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Sample records for saliva peptide p-c

  1. The pH changes of artificial saliva after interaction with oral of artificial saliva after interaction with oral micropathogen

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    Basri A. Gani

    2012-12-01

    Full Text Available Backgorund: Saliva contains several protein elements, exocrine proteins and antibodies, such as lactoferrin, sIgA, peroxidase, albumin, polypeptides, and oligopeptides that contribute to the defense of oral mucosa and dental pellicle to prevent infection caused by oral micropathogen, such as Candida albicans, Streptococcus mutans and Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans. Those micropathogens have a role to change salivary pH as an indicator of oral disease activities. Purpose: This study was aimed to analyze the changes of artificial saliva pH after interaction with S. mutans, C. albicans, and A. Actinomycetemcpmitans. Methods: The materials used in this study consist of S. mutans (ATCC 31987, C. albicans (ATCC 10231, A. actinomycetemcomitans (ATTC 702 358, and artificial saliva. To examine the pH changes of artificial saliva, those three microbiotas were cultured and incubated for 24 hours. Results: The results showed that the interactions of S. mutans, C. albicans, and A. actinomycetemcomitans in the artificial saliva can change the salivary on neutral. There were no significant difference with the control treatment salivary pH 4, 5, 6, 8, and 9 (p>0.05. Similarly, there was also no significant difference when those three microorganism interacted each other in the artificial saliva (p<0.05. Conclusion: It can be concluded that the biological activity of S. mutans, C. albicans, and A. actinomycetemcomitans in artificial saliva can change the salivary pH into neutral. It indicates that those microbiotas mutually supported and cooperated in influencing the biological cycle of the oral cavity with salivary pH as an indicator.Latar belakang: Saliva merupakan cairan eksokrin yang mengandung unsur protein dan antibodi seperti sIgA laktoferin peroksidase, albumin, polipeptida dan oligopeptida yang berperan pada pertahanan mukosa rongga mulut dan gigi guna mencegah infeksi oral mikropatogen seperti C. albicans, S. mutans, dan

  2. Identities of P2 and P3 Residues of H-2Kb-Bound Peptides Determine Mouse Ly49C Recognition.

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    Elsa A Marquez

    Full Text Available Ly49 receptors can be peptide selective in their recognition of MHC-I-peptide complexes, affording them a level of discrimination beyond detecting the presence or absence of specific MHC-I allele products. Despite this ability, little is understood regarding the properties that enable some peptides, when bound to MHC-I molecules, to support Ly49 recognition, but not others. Using RMA-S target cells expressing MHC-I molecules loaded with individual peptides and effector cells expressing the ectodomain of the inhibitory Ly49C receptor, we found that two adjacent amino acid residues, P2 and P3, both buried in the peptide binding groove of H-2Kb, determine mouse Ly49C specificity. If both are aliphatic residues, this is supportive. Whereas, small amino acids at P2 and aromatic amino acids at the P3 auxiliary anchor residue are detrimental to Ly49C recognition. These results resemble those with a rat Ly49 where the identity of a peptide anchor residue determines recognition, suggesting that dependence on specific peptide residues buried in the MHC-I peptide-binding groove may be fundamental to Ly49 peptide selectivity and recognition.

  3. CHROMOGRANIN A DETECTION IN SALIVA OF TYPE 2 DIABETES PATIENTS

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    Martine Soell

    2010-02-01

    Full Text Available Chromogranin A is present in secretion granules of nerve, endocrine and immune cells and is a precursor of several peptides with antibacterial and antifungal properties at micromolar concentrations.Our aim in this prospective, double blind study, was to determine the expression of chromogranin A and its peptides at protein level in saliva of type 2 diabetic patients and thereby to obtain a new non-invasive diagnostic means for the future.Saliva was taken from 30 type 2 diabetic patients and 30 healthy individuals at the same time interval in the morning without any oral stimuli. Circadianic periodics in protein productions have been avoided. The presence of chromogranin A and its derived peptides was determined in whole saliva, after centrifugation at 40C for 12 min at 14 000 rpm, by SDS-PAGE electrophoresis and Immunoblotting (Western Blot. To ensure same protein concentrations Bradford protein quantification assay has been performed before.For the first time, we have determined an overexpression of chromogranin A in saliva of diabetic patients in 100% of the individuals.Chromogranin A, a circulating biomarker for epithelial tumours, is also overexpressed in saliva of type 2 diabetic patients. To confirm our results, more studies with a large amount of patients is necessary.

  4. Chromogranin A Detection in Saliva of Type 2 Diabetes Patients

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    Martine Soell

    2010-02-01

    Full Text Available Chromogranin A is present in secretion granules of nerve, endocrine and immune cells and is a precursor of several peptides with antibacterial and antifungal properties at micromolar concentrations.Our aim in this prospective, double blind study, was to determine the expression of chromogranin A and its peptides at protein level in saliva of type 2 diabetic patients and thereby to obtain a new non-invasive diagnostic means for the future.Saliva was taken from 30 type 2 diabetic patients and 30 healthy individuals at the same time interval in the morning without any oral stimuli. Circadianic periodics in protein productions have been avoided. The presence of chromogranin A and its derived peptides was determined in whole saliva, after centrifugation at 4°C for 12 min at 14 000 rpm, by SDS-PAGE electrophoresis and Immunoblotting (Western Blot. To ensure same protein concentrations Bradford protein quantification assay has been performed before.For the first time, we have determined an overexpression of chromogranin A in saliva of diabetic patients in 100% of the individuals.Chromogranin A, a circulating biomarker for epithelial tumours, is also overexpressed in saliva of type 2 diabetic patients. To confirm our results, more studies with a large amount of patients is necessary.

  5. Pengaruh Stimulus Pengunyahan dan Pengecapan Terhadap Kecepatan Aliran dan pH Saliva

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    Hj. Edeh Rolette Haroen

    2015-09-01

    Full Text Available The aim of the research were to describe how salivary flow rate and pH vary with time during use of chewing and gustatory stimulation. Fifty young adult subjects collected unstimulated saliva by spitting method, and then collected stimulated saliva by chewing paraffin wax, and a few drops of citric acid are usually placed on the subject’s tongue. The mean of saliva flow rate that unstimulated: 0.50 cc/minute; stimulated saliva by chewing paraffin wax: 1.57 cc/minute, and drops of citric acid stimulation showed that saliva flow rate: 2.98 cc/minute; and pH saliva that unstimulated 6.39; stimulated saliva by chewing paraffin wax 7.2; and stimulated saliva by citric acid: 7.55. Statistical paired t test showed that t lower than t table. The conclusion of the research showed that there were significant influences in the unstimulated salivary flow rates and pH with stimulated saliva elicited by chewing and gustatory stimulation.

  6. Estudo preliminar do potencial cariogênico de preparações doces da merenda escolar através do pH da saliva Preliminary study of cariogenic potential of sweet school meal preparations through saliva pH

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    Cristiane Pinheiro Lázaro

    1999-12-01

    Full Text Available Objetivou-se determinar a curva de pH da saliva após o consumo de preparações doces, relacionar as variáveis teor de glicídios e pH, e discutir dados sobre o uso de preparações doces na merenda escolar do município de Niterói, RJ, com diferentes teores de glicídios e diferentes consistências, com crianças em idade pré-escolar e escolar, atendidas no Departamento de Odontologia Clínica da Faculdade de Odontologia da Universidade Federal Fluminense. Para os testes, foram selecionadas 52 crianças eutróficas, sadias, sem uso de medicamentos que alterassem o fluxo e a composição da saliva e com baixo e alto risco de cárie. Neste grupo, 54% apresentavam idade de 7 a 10 anos. Foi medido o pH da saliva, com fitas indicadoras, antes e após o consumo destas preparações, em um intervalo de 10, 20, 40 e 60 minutos. O tratamento estatístico foi realizado através do "software" Epi Info e foram realizados os testes de Qui-Quadrado e "t" de Student. Nenhuma das curvas de pH da saliva, para todas as preparações testadas e grupos de risco atingiram valor de pH considerado crítico (5,3 - 5,5. As curvas de pH médio da saliva, para todas as preparações testadas, oscilaram em valores de pH mais elevados para o grupo de baixo risco de cárie e em valores mais baixos para o grupo de alto risco de cárie. As curvas de pH médio da saliva do grupo de baixo risco demonstraram queda mais lenta e recuperação mais tardia, enquanto que no grupo de alto risco a queda e a recuperação do pH foram mais imediatas. A consistência das preparações foi preponderante sobre o teor de glicídios. Recomenda-se que o planejamento da merenda escolar leve em consideração o efeito da consistência das preparações doces servidas.The aim of this study was to establish the saliva pH curve in response to sweet foods, with different carbohydrate rates and different consistencies, frequently found in the school meal program in Niterói, Rio de Janeiro

  7. Saliva as a tool for monitoring steroid, peptide and immune markers in sport and exercise science.

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    Papacosta, Elena; Nassis, George P

    2011-09-01

    This paper discusses the use of saliva analysis as a tool for monitoring steroid, peptide, and immune markers of sports training. Salivary gland physiology, regarding the regulation and stimulation of saliva secretion, as well as methodological issues including saliva collection, storage and analysis are addressed in this paper. The effects of exercise on saliva composition are then considered. Exercise elicits changes in salivary levels of steroid hormones, immunoglobulins, antimicrobial proteins and enzymes. Cortisol, testosterone and dehydroepiandrosterone can be assessed in saliva, providing a non-invasive option to assess the catabolic and anabolic effects of exercise. Validation studies using blood and salivary measures of steroid hormones are addressed in this paper. Effects of acute exercise and training on salivary immunoglobulins (SIgA, SIgM, SIgG) and salivary antimicrobial proteins, including α-amylase, lysozyme and lactoferrin, are also discussed. Analysis of cortisol and testosterone in saliva may help detect the onset of non-functional overreaching and subsequently may help to prevent the development of overtraining syndrome. Assessment of salivary immunoglobulins and antimicrobial proteins has been shown to successfully represent the effects of exercise on mucosal immunity. Increases in SIgA and antimicrobial proteins concentration and/or secretion rate are associated with acute exercise whereas conversely, decreases have been reported in athletes over a training season leaving the athlete susceptible for upper respiratory tract infections. The measurement of physiological biomarkers in whole saliva can provide a significant tool for assessing the immunological and endocrinological status associated with exercise and training. Copyright © 2011 Sports Medicine Australia. Published by Elsevier Ltd. All rights reserved.

  8. Identifikasi epitop dari Streptococcus mutans terhadap sekretori Imunoglobulin A saliva (The identification of Streptococcus mutans epitopes to secretory Immunoglobulin A saliva

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    Anita Yuliati

    2005-09-01

    Full Text Available S. mutans is one of the etiology agent of dental caries, these bacteria have a surface protein of about 185 kDa named Ag I/II. The secretory of sIgA saliva to Ag I/II of S.mutans has shown to be able to prevent colonization in human oral cavity. Peptides derived from the 824 to 853 residues of the P region of antigen I/II S. mutans related to the pathogenesis of dental caries. The aim of this study was to identify the overlapping sequence of amino acids (epitope derived from the 624 to 853 residues of P of antigen I/II S. mutans to sIgA saliva on caries and caries-free subject in a observational cross sectional study. The P region of antigen I/II S.mutans was cut into 22 peptides of 9 mer sequences with an overlapping of 8 mer and an offset of 1 mer, synthesized on polyethylene pins and tested for the reactivity with an ELISA indirect method to sIgA saliva on caries and caries-free subject. The results of this study showed that amino acid sequences with TPPVKP (832–837 and TAPTKPTY (838–845 were reactive to sIgA saliva on caries and caries-free subject. The conclusion of this study was that the overlapping common sequence of amino acid (epitopes corresponding to TPPVKP (832–837 and TAPTKPTY (838–845 was identified as caries marker epitopes in human.

  9. Amperometric micro pH measurements in oxygenated saliva.

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    Chaisiwamongkhol, Korbua; Batchelor-McAuley, Christopher; Compton, Richard G

    2017-07-24

    An amperometric micro pH sensor has been developed based on the chemical oxidation of carbon fibre surfaces (diameter of 9 μm and length of ca. 1 mm) to enhance the population of surface quinone groups for the measurement of salivary pH. The pH analysis utilises the electrochemically reversible two-electron, two-proton behaviour of surface quinone groups on the micro-wire electrodes. A Nernstian response is observed across the pH range 2-8 which is the pH range of many biological fluids. We highlight the measurement of pH in small volumes of biological fluids without the need for oxygen removal and specifically the micro pH electrode is examined by measuring the pH of commercial synthetic saliva and authentic human saliva samples. The results correspond well with those obtained by using commercial glass pH electrodes on large volume samples.

  10. [Concentration of calcium ions in the saliva and the value of the pH of the saliva in female and male smokers].

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    Nakonieczna-Rudnicka, Marta; Bachanek, Teresa; Rogowska, Wanda

    2009-01-01

    Dental decay is a pathological process of extrasomatic origin which leads to demineralization and proteolytic degradation of hard surfaces of a tooth susceptible to this disease. Saliva composition, including calcium ion concentration and its pH value, is of importance in the development of the carious process. Tobacco smoke contains toxic compounds which negatively influence oral health. The aim of the study was evaluation of the selected saliva components: protein concentration, Ca2+ concentration, pH value both in male and female smokers. The investigated group included 65 patients reporting for the treatment to the Department of Conservative Dentistry of Medical University in Lublin. In the investigated group male smokers constituted 15.38%, female smokers--20.00%, male nicotine abstinents 21.54% and female nicotine abstinent 43.08%. The study included both survey examinations of patients and biochemical examinations of the saliva. Mixed, non-stimulated saliva was used as a material for biochemical examinations. Ca2+ concentration and pH of the saliva were assayed with the use of Rapidlab 348 analyzer. Protein in the saliva was assayed with calorimetric method according to Lowry. Saliva was collected from smokers 10-120 minutes after smoking of several cigarettes. It was stated that Ca2+ and protein concentration as well as pH of the saliva were not correlated with sex and cigarette smoking or non-smoking.

  11. Clinical significance of determination of serum C-peptide levels

    International Nuclear Information System (INIS)

    Wang Guohong; Xu Ruiji; Zhang Zhongshu; Wang Xiaoji

    2006-01-01

    Objective: To study the clinical meanings of changes of serum C-peptide levels and insulin/C-peptide ratio. Methods: Serum insulin and C-peptide levels were determined with RIA in 171 patients with DM-2 of all ages (31-50, n= 50, 51-60, n=60, over 60, n=61) and 50 patients with renal insufficiency. The insulin/C-peptide ratio were calculated. Results: The serum C-peptide and insulin levels in patients with renal insufficiency were significantly higher than those in diabetics of all age groups and the insulin/C-peptide ratio were significantly lower than those in diabetics (P 0.05), but the serum C-peptide levels increased as the age of patients increased with decrease of insulin/C-peptide ratio (P<0.01). Conclusion: Abnormal changes of C-peptide levels and insulin/C-peptide ratio in diabetics (the age-factor corrected) might reflect renal dysfunction. (authors)

  12. Pengaruh (pH Saliva terhadap Terjadinya Karies Gigi pada Anak Usia Prasekolah

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    Made Ayu Lely

    2017-12-01

    Full Text Available AbstractDental caries is a multifactorial process that occurs through the interaction between teeth and saliva as host, the bacteria in the oral cavity, as well as easily fermented foods. Saliva is one of the factors that have a major influence on the severity of dental caries. The aim of this study was to determine the relationship of salivary pH with dental caries among 564 preschool-age children in DIY Province and Banten Province. The results showed that pH levels of the preschool-age children’saliva are more than 75% basic ranging between 6.8 to 8.0 and the highest levels are in Serang District. Index def-t in Serang District is highest (8.83 and the lowest one is in Yogyakarta City (4.97. The mean number of cavities/ decay more than missing teeth or filling teeth. The study indicates that the acidity of saliva among preschool children in the two provinces is not associated with the occurrence of dental caries.It is more likely due to the habit of drinking sweet milk or eating sticky foods.Key words: thepH of saliva, dental caries,sweet food, sticky foods, preschool childrenageAbstrakKaries gigi merupakan proses multifaktor yang terjadi melalui interaksi antara gigi dan saliva sebagai pejamu, bakteri didalam rongga mulut, serta makanan yang mudah difermentasikan. Saliva merupakan salah satu faktor yang mempunyai pengaruh besar terhadap keparahan karies gigi. Tujuan dari penelitian ini adalah untuk mengetahui pengaruh pH saliva terhadap terjadinya karies gigi pada anak usia prasekolah. Penelitian dilakukan secara potong lintang pada 564 orang anak usia prasekolah di Provinsi Daerah Istimewa Yogyakarta (Provinsi DIY dan Provinsi Banten. Hasil penelitian menunjukkan bahwa derajat keasaman (pH saliva pada anak-anak usia prasekolah lebih dari 75% bersifat basa berkisar antara 6,8-8,0 dan tertinggi di Kabupaten Serang. Indeks def-t tertinggi 8,83 di Kabupaten Serang dan yang terendah 4,97 di Kotamadya Yogyakarta. Rerata jumlah gigi berlubang

  13. Preliminary findings on the correlation of saliva pH, buffering ...

    African Journals Online (AJOL)

    The aim of the present comparative study was to compare some salivary characteristics between exclusive waterpipe smokers (EWPS) and non-smokers. 72 males (36 EWPS) were recruited. The volume of stimulated saliva was determined and divided by the duration of saliva collection. The pH was measured directly ...

  14. Correlations between fasting plasma C-peptide, glucagon-stimulated plasma C-peptide, and urinary C-peptide in insulin-treated diabetics

    DEFF Research Database (Denmark)

    Gjessing, H J; Matzen, L E; Frøland, A

    1987-01-01

    This study correlated fasting plasma C-peptide (CP), plasma CP 6 min after stimulation with 1 mg glucagon i.v., and the mean of three 24-h urinary excretions of C-peptide (UCP)/creatinine in 132 insulin-treated diabetics. Patients were divided into three groups: group 1, stimulated CP less than 0.......06 nM (n = 51); group 2, stimulated CP 0.06-0.60 nM (n = 48); and group 3, stimulated CP greater than 0.60 nM (n = 33). In all patients fasting CP was closely correlated to stimulated CP (r = .988, P less than .001), whereas the correlations between UCP and both fasting CP (r = .904, P less than .001......) and stimulated CP r = .902, P less than .001) were slightly less pronounced. The associations between UCP and both fasting CP (r = .716, P less than .001) and stimulated CP (r = .731, P less than .001) were modest in group 2, and even more so in group 3 (r = .557, P less than .001 and r = .641, P less than .001...

  15. The proteome of human saliva

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    Griffin, Timothy J.

    2013-05-01

    Human saliva holds tremendous potential for transforming disease and health diagnostics given its richness of molecular information and non-invasive collection. Enumerating its molecular constituents is an important first step towards reaching this potential. Among the molecules in saliva, proteins and peptides arguably have the most value: they can directly indicate biochemical functions linked to a health condition/disease state, and they are attractive targets for biomarker assay development. However, cataloging and defining the human salivary proteome is challenging given the dynamic, chemically heterogeneous and complex nature of the system. In addition, the overall human saliva proteome is composed of several "sub-proteomes" which include: intact full length proteins, proteins carrying post-translational modifications (PTMs), low molecular weight peptides, and the metaproteome, derived from protein products from nonhuman organisms (e.g. microbes) present in the oral cavity. Presented here will be a summary of communal efforts to meet the challenge of characterizing the multifaceted saliva proteome, focusing on the use of mass spectrometry as the proteomic technology of choice. Implications of these efforts to characterize the salivary proteome in the context of disease diagnostics will also be discussed.

  16. Electrochemical behavior and pH stability of artificial salivas for corrosion tests.

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    Queiroz, Gláucia Maria Oliveira de; Silva, Leandro Freitas; Ferreira, José Tarcísio Lima; Gomes, José Antônio da Cunha P; Sathler, Lúcio

    2007-01-01

    It is assumed that the compositions of artificial salivas are similar to that of human saliva. However, the use of solutions with different compositions in in vitro corrosion studies can lead dissimilar electrolytes to exhibit dissimilar corrosivity and electrochemical stability. This study evaluated four artificial salivas as regards pH stability with time, redox potentials and the polarization response of an inert platinum electrode. The tested solutions were: SAGF medium, Mondelli artificial saliva, UFRJ artificial saliva (prepared at the School of Pharmacy, Federal University of Rio de Janeiro, RJ, Brazil) and USP-RP artificial saliva (prepared at the School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, SP, Brazil). It was observed that pH variations were less than 1 unit during a 50-hour test. The SAGF medium, and the UFRJ and USP-RP solutions exhibited more oxidizing characteristics, whereas the Mondelli solution presented reducing properties. Anodic polarization revealed oxidation of the evaluated electrolytes at potentials below +600 mV SCE. It was observed that the UFRJ and USP-RP solutions presented more intense oxidation and reduction processes as compared to the Mondelli and SAGF solutions.

  17. Flow rate, pH and calcium concentration of saliva of children and adolescents with type 1 diabetes mellitus.

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    Moreira, A R; Passos, I A; Sampaio, F C; Soares, M S M; Oliveira, R J

    2009-08-01

    Alterations in salivary parameters may increase the caries risk in diabetic children, but, contradictory data on this issue have been reported. The aims of this study were to compare salivary parameters (flow rate, pH and calcium concentration) between healthy and type 1 diabetes mellitus (T1DM) individuals. The sample consisted of 7- to 18-year-old individuals divided into two groups: 30 subjects with T1DM (group A) and 30 healthy control subjects (group B). Fasting glucose levels were determined. Unstimulated and stimulated saliva was collected. The pH of unstimulated saliva was measured with paper strips and an electrode. Calcium concentrations in stimulated saliva were determined with a selective electrode. Group A individuals had inadequate blood glucose control (HbA(1C) >9%), with means +/- SD unstimulated salivary flow rate of 0.15 +/- 0.1 mL/min compared to 0.36 +/- 0.2 mL/min for group B (P salivary flow rate was similar by both groups and above 2.0 mL/min. Saliva pH was 6.0 +/- 0.8 for group A and significantly different from 7.0 +/- 0.6 for group B (P Salivary calcium was 14.7 +/- 8.1 mg/L for group A and significantly higher than 9.9 +/- 6.4 mg/L for group B (P salivary parameters favoring caries such as low saliva pH and unstimulated salivary flow rate were observed in T1DM individuals.

  18. Flow rate, pH and calcium concentration of saliva of children and adolescents with type 1 diabetes mellitus

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    A.R. Moreira

    2009-08-01

    Full Text Available Alterations in salivary parameters may increase the caries risk in diabetic children, but, contradictory data on this issue have been reported. The aims of this study were to compare salivary parameters (flow rate, pH and calcium concentration between healthy and type 1 diabetes mellitus (T1DM individuals. The sample consisted of 7- to 18-year-old individuals divided into two groups: 30 subjects with T1DM (group A and 30 healthy control subjects (group B. Fasting glucose levels were determined. Unstimulated and stimulated saliva was collected. The pH of unstimulated saliva was measured with paper strips and an electrode. Calcium concentrations in stimulated saliva were determined with a selective electrode. Group A individuals had inadequate blood glucose control (HbA1C >9%, with means ± SD unstimulated salivary flow rate of 0.15 ± 0.1 mL/min compared to 0.36 ± 0.2 mL/min for group B (P < 0.01. Stimulated salivary flow rate was similar by both groups and above 2.0 mL/min. Saliva pH was 6.0 ± 0.8 for group A and significantly different from 7.0 ± 0.6 for group B (P < 0.01. Salivary calcium was 14.7 ± 8.1 mg/L for group A and significantly higher than 9.9 ± 6.4 mg/L for group B (P < 0.01. Except for elevated calcium concentrations in saliva, salivary parameters favoring caries such as low saliva pH and unstimulated salivary flow rate were observed in T1DM individuals.

  19. Electrochemical behavior and pH stability of artificial salivas for corrosion tests Comportamento eletroquímico e estabilidade de pH de salivas artificiais para testes de corrosão

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    Gláucia Maria Oliveira de Queiroz

    2007-09-01

    Full Text Available It is assumed that the compositions of artificial salivas are similar to that of human saliva. However, the use of solutions with different compositions in in vitro corrosion studies can lead dissimilar electrolytes to exhibit dissimilar corrosivity and electrochemical stability. This study evaluated four artificial salivas as regards pH stability with time, redox potentials and the polarization response of an inert platinum electrode. The tested solutions were: SAGF medium, Mondelli artificial saliva, UFRJ artificial saliva (prepared at the School of Pharmacy, Federal University of Rio de Janeiro, RJ, Brazil and USP-RP artificial saliva (prepared at the School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, SP, Brazil. It was observed that pH variations were less than 1 unit during a 50-hour test. The SAGF medium, and the UFRJ and USP-RP solutions exhibited more oxidizing characteristics, whereas the Mondelli solution presented reducing properties. Anodic polarization revealed oxidation of the evaluated electrolytes at potentials below +600 mV SCE. It was observed that the UFRJ and USP-RP solutions presented more intense oxidation and reduction processes as compared to the Mondelli and SAGF solutions.Admite-se que as composições das salivas artificiais são semelhantes àquela da saliva humana. A utilização de soluções de composições distintas em estudos de corrosão in vitro, entretanto, pode fazer com que eletrólitos diferentes exibam diferenças no processo corrosivo e na estabilidade eletroquímica. Este estudo avaliou quatro salivas artificiais em relação a estabilidade do pH em função do tempo, potencial redox e resposta à polarização de um eletrodo de platina inerte. As soluções testadas foram: meio SAGF, saliva artificial Mondelli, saliva artificial UFRJ (preparada pela Faculdade de Farmácia da Universidade Federal do Rio de Janeiro, RJ, Brasil e saliva artificial USP-RP (preparada pela

  20. Comparing Properties (Concentration, PH and mutans streptococcus Saliva in Both Status Resting Saliva and Stimulated Saliva in Preschoolers of Kerman city

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    Elham Farokh-Gisour,

    2016-08-01

    Full Text Available This paper aimed to compare the characteristics (concentration, PH and mutans streptococcus saliva in both status resting saliva and stimulated saliva in preschoolers of Kerman city. In this study, 100 children aged 5 years among patients admitted to the pediatric ward of Kerman dental school and dental offices, some experts in Kerman dental school participated. Resting and stimulated saliva (after chewing oral paraffin children collected and in concentrations, PH and the amount of mutans streptococcus was measured. Mc Nemar test to compare the frequency of positive and negative cultures before and after stimulation as well as paired t-test to compare the saliva pH and concentration of not stimulated saliva and stimulated saliva in two modes was used. The significance level was set less than 0.05.The mean resting salivary osmolality of the population: 30.42 ± 87.41 and the average salivary osmolality of the total population were 79.81. Osmolality differences in saliva before and after stimulation with each other was significant (p = 0.009, paired t-test. The mean of resting saliva in the total population PH 0.45 ± 7.78 and the average PH stimulated saliva in the total population was 8.22 and the difference before and after each significant (p = 0.02, paired t-test. In mutans streptococcus in test samples in all 71 patients (71% positive test and 29 patients (29% had a negative test that number of positive cultures are equal before and after stimulation of saliva and thus the difference between the two groups (p> 0.05 was observed. In terms of comparing the properties of resting and stimulated saliva can conclude that salivary stimulated PH was significantly higher than resting saliva. While stimulated saliva osmolality was significantly less than resting saliva and the frequency of positive test mutans streptococcus in saliva before and after stimulation had no significant difference (p> 0.05. This means that test results on samples of mutans

  1. Proinsulin C-peptide interferes with insulin fibril formation

    International Nuclear Information System (INIS)

    Landreh, Michael; Stukenborg, Jan-Bernd; Willander, Hanna; Söder, Olle; Johansson, Jan; Jörnvall, Hans

    2012-01-01

    Highlights: ► Insulin and C-peptide can interact under insulin fibril forming conditions. ► C-peptide is incorporated into insulin aggregates and alters aggregation lag time. ► C-peptide changes insulin fibril morphology and affects backbone accessibility. ► C-peptide may be a regulator of fibril formation by β-cell granule proteins. -- Abstract: Insulin aggregation can prevent rapid insulin uptake and cause localized amyloidosis in the treatment of type-1 diabetes. In this study, we investigated the effect of C-peptide, the 31-residue peptide cleaved from proinsulin, on insulin fibrillation at optimal conditions for fibrillation. This is at low pH and high concentration, when the fibrils formed are regular and extended. We report that C-peptide then modulates the insulin aggregation lag time and profoundly changes the fibril appearance, to rounded clumps of short fibrils, which, however, still are Thioflavine T-positive. Electrospray ionization mass spectrometry also indicates that C-peptide interacts with aggregating insulin and is incorporated into the aggregates. Hydrogen/deuterium exchange mass spectrometry further reveals reduced backbone accessibility in insulin aggregates formed in the presence of C-peptide. Combined, these effects are similar to those of C-peptide on islet amyloid polypeptide fibrillation and suggest that C-peptide has a general ability to interact with amyloidogenic proteins from pancreatic β-cell granules. Considering the concentrations, these peptide interactions should be relevant also during physiological secretion, and even so at special sites post-secretory or under insulin treatment conditions in vivo.

  2. Proinsulin C-peptide interferes with insulin fibril formation

    Energy Technology Data Exchange (ETDEWEB)

    Landreh, Michael [Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm (Sweden); Stukenborg, Jan-Bernd [Department of Women' s and Children' s Health, Astrid Lindgren Children' s Hospital, Pediatric Endocrinology Unit, Karolinska Institutet and University Hospital, S-17176 Stockholm (Sweden); Willander, Hanna [KI-Alzheimer' s Disease Research Center, NVS Department, Karolinska Institutet, S-141 86 Stockholm (Sweden); Soeder, Olle [Department of Women' s and Children' s Health, Astrid Lindgren Children' s Hospital, Pediatric Endocrinology Unit, Karolinska Institutet and University Hospital, S-17176 Stockholm (Sweden); Johansson, Jan [KI-Alzheimer' s Disease Research Center, NVS Department, Karolinska Institutet, S-141 86 Stockholm (Sweden); Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, S-751 23 Uppsala (Sweden); Joernvall, Hans, E-mail: Hans.Jornvall@ki.se [Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm (Sweden)

    2012-02-17

    Highlights: Black-Right-Pointing-Pointer Insulin and C-peptide can interact under insulin fibril forming conditions. Black-Right-Pointing-Pointer C-peptide is incorporated into insulin aggregates and alters aggregation lag time. Black-Right-Pointing-Pointer C-peptide changes insulin fibril morphology and affects backbone accessibility. Black-Right-Pointing-Pointer C-peptide may be a regulator of fibril formation by {beta}-cell granule proteins. -- Abstract: Insulin aggregation can prevent rapid insulin uptake and cause localized amyloidosis in the treatment of type-1 diabetes. In this study, we investigated the effect of C-peptide, the 31-residue peptide cleaved from proinsulin, on insulin fibrillation at optimal conditions for fibrillation. This is at low pH and high concentration, when the fibrils formed are regular and extended. We report that C-peptide then modulates the insulin aggregation lag time and profoundly changes the fibril appearance, to rounded clumps of short fibrils, which, however, still are Thioflavine T-positive. Electrospray ionization mass spectrometry also indicates that C-peptide interacts with aggregating insulin and is incorporated into the aggregates. Hydrogen/deuterium exchange mass spectrometry further reveals reduced backbone accessibility in insulin aggregates formed in the presence of C-peptide. Combined, these effects are similar to those of C-peptide on islet amyloid polypeptide fibrillation and suggest that C-peptide has a general ability to interact with amyloidogenic proteins from pancreatic {beta}-cell granules. Considering the concentrations, these peptide interactions should be relevant also during physiological secretion, and even so at special sites post-secretory or under insulin treatment conditions in vivo.

  3. Insulin C-peptide test

    Science.gov (United States)

    C-peptide ... the test depends on the reason for the C-peptide measurement. Ask your health care provider if ... C-peptide is measured to tell the difference between insulin the body produces and insulin someone injects ...

  4. Fasting plasma C-peptide, glucagon stimulated plasma C-peptide, and urinary C-peptide in relation to clinical type of diabetes

    DEFF Research Database (Denmark)

    Gjessing, H J; Matzen, L E; Faber, O K

    1989-01-01

    with a fasting plasma C-peptide value less than 0.20 nmol/l, a glucagon stimulated plasma C-peptide value less than 0.32 nmol/l, and a urinary C-peptide value less than 3.1 nmol/l, or less than 0.54 nmol/mmol creatinine/24 h, or less than 5.4 nmol/24 h mainly were Type 1 diabetic patients; while patients with C...

  5. Time-related Changes in pH, Buffering Capacity and Phosphate and Urea Concentration of Stimulated Saliva.

    Science.gov (United States)

    Vuletic, Lea; Peros, Kristina; Spalj, Stjepan; Rogic, Dunja; Alajbeg, Ivan

    2014-01-01

    To quantify changes in pH, buffering capacity and hydrogen carbonate, phosphate, protein and urea concentrations of stimulated saliva which occur during a 30-min measurement delay after saliva collection. The correlation between time-related chemical changes and changes of salivary pH and buffering capacity was assessed in order to explain the observed changes in salivary pH and buffering capacity. Stimulated saliva samples were collected from 30 volunteers after inducing salivation by chewing a piece of parafilm. Measurements of salivary variables were made immediately after saliva collection and again 30 min later, during which time the specimens were exposed to the atmosphere in collection cups at room temperature. Postponement of measurements resulted in a significant increase in pH and a significant decrease of buffering capacity, phosphate and urea concentration. The results suggest that the time-related pH increase could primarily be attributed to loss of dissolved carbon dioxide from saliva, and confirm the importance of hydrogen carbonate in the neutralisation of hydrogen ions, but they do not support the principle of catalysed phase-buffering for the hydrogen carbonate buffer system in saliva. A decrease in phosphate and urea concentration affects salivary buffering capacity. This study emphasises the importance of the standardisation of measurement time when measuring salivary pH, buffering capacity, phosphate and urea concentrations following the collection of saliva in order to obtain comparable results. It also provides a partial explanation of the mechanisms underlying the observed changes of pH and buffering capacity over time.

  6. Use of Saliva Biomarkers to Monitor Efficacy of Vitamin C in Exercise-Induced Oxidative Stress

    Directory of Open Access Journals (Sweden)

    Levi W. Evans

    2017-01-01

    Full Text Available Saliva is easily obtainable for medical research and requires little effort or training for collection. Because saliva contains a variety of biological compounds, including vitamin C, malondialdehyde, amylase, and proteomes, it has been successfully used as a biospecimen for the reflection of health status. A popular topic of discussion in medical research is the potential association between oxidative stress and negative outcomes. Systemic biomarkers that represent oxidative stress can be found in saliva. It is unclear, however, if saliva is an accurate biospecimen as is blood and/or plasma. Exercise can induce oxidative stress, resulting in a trend of antioxidant supplementation to combat its assumed detriments. Vitamin C is a popular antioxidant supplement in the realm of sports and exercise. One potential avenue for evaluating exercise induced oxidative stress is through assessment of biomarkers like vitamin C and malondialdehyde in saliva. At present, limited research has been done in this area. The current state of research involving exercise-induced oxidative stress, salivary biomarkers, and vitamin C supplementation is reviewed in this article.

  7. Increase in acrolein-conjugated immunoglobulins in saliva from patients with primary Sjögren's syndrome.

    Science.gov (United States)

    Hirose, Tadao; Saiki, Ryotaro; Uemura, Takeshi; Suzuki, Takehiro; Dohmae, Naoshi; Ito, Satoshi; Takahashi, Hoyu; Ishii, Itsuko; Toida, Toshihiko; Kashiwagi, Keiko; Igarashi, Kazuei

    2015-10-23

    We previously reported that the level of protein-conjugated acrolein (PC-Acro), a marker of cell or tissue damage, was increased in saliva from patients with primary Sjögren's syndrome (pSS), and that the level of PC-Acro was well correlated with the severity of pSS. Acrolein-conjugated immunoglobulins were measured in saliva from pSS patients. The activities of autoantibodies recognizing Sjögren's syndrome SSA (Ro) and SSB (La) proteins in saliva from pSS patients were approximately 3- to 5-fold higher than those from control subjects. We also found that autoantibody activities recognizing SSA (Ro) and SSB (La) proteins increased after acrolein treatment of saliva from control subjects. When an antibody against human serum albumin was treated with acrolein, the ability to recognize albumin was reduced but the ability to recognize other proteins was increased. Twenty-four and eleven kinds of acrolein-conjugated amino acids were found at the variable and constant regions of peptides, respectively, obtained from the immunoglobulins in saliva from pSS patients. The altered recognition patterns of immunoglobulins due to acrolein conjugation are at least partially involved in autoimmune diseases. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Isolation and Analytical Characterization of Local Malaysian Leech Saliva

    Directory of Open Access Journals (Sweden)

    Mohamed Alaama

    2011-12-01

    Full Text Available Leech saliva contains biologically active compounds that are mainly proteins and peptides. In this study a modified and smooth extraction method of saliva was used without leeches' scarification. UV and Bradford Assay protein methods showed that the saliva extract contains high concentrations of proteins. RP-HPLC chromatogram revealed that more than 30 different peaks were observed in leech saliva extract. Gel electrophoresis revealed the existence of proteins and peptides with different molecular weights. The gel showed up to 25 different bands. Comparison of gel electrophoresis data with protein database revealed the closeness of four molecular weights to known proteins from Hirudinaria leech family. Other proteins detected by gel electrophoresis may be related to completely new biologically active proteins and peptides in the saliva extract or to a modification (isoforms of the existing ones or finally to a mixture of both.ABSTRAK: Air liur pacat secara biologinya mengandungi sebahagian besar campuran aktif protein dan peptida. Dalam kajian ini, kaedah pengestrakan air liur pacat yang telah diubah suai digunakan tanpa perlu membunuh pacat. Kaedah protein Cerakin UV dan Bradford menunjukkan air liur pacat yang diekstrak mengandungi konsentrasi protein yang tinggi. Kromatogram RP-HPLC memperlihatkan lebih daripada 30 puncak berbeza diperolehi semasa air liur pacat diekstrak. Gel elektroforesis memperlihatkan kewujudan protein dan peptida dengan berat molekul yang berbeza. Gel menunjukkan hingga 25 jalur yang berbeza. Perbandingan data menggunakan gel elektroforesis seiring dengan pangkalan data protein memperlihatkan persamaan empat berat molekul, dengan protein yang yang dikenali daripada keluarga pacat Hirudinaria. Jenis protein lain yang dikesan dengan menggunakan gel elektrofosis mungkin juga berkait secara biologinya dengan protein dan peptida aktif yang baru, dalam ekstrak air liur atau pengubahsuaian (beberapa jenis yang berbeza daripada

  9. Association between low C-peptide and fragility fractures in postmenopausal women without diabetes.

    Science.gov (United States)

    Ferro, Y; Russo, C; Russo, D; Gazzaruso, C; Coppola, A; Gallotti, P; Zambianchi, V; Fodaro, M; Romeo, S; Galliera, E; Marazzi, M G; Romanelli, M M C; Giannini, S; Pujia, A; Montalcini, T

    2017-10-01

    C-peptide has been shown to exert several, previously unknown, biological effects. A recent cross-sectional study demonstrated an association between low C-peptide serum levels and low lumbar bone density of postmenopausal women not affected by diabetes. To date, very little research attention has been directed toward the association between C-peptide and osteoporotic fractures. To contribute toward filling this gap, we investigated the association between C-peptide and fractures in postmenopausal women. A cohort of 133 non-diabetic postmenopausal women with and without a history of fractures was evaluated in this cross-sectional investigation. Standardized interviews were performed to gather information on the patients' fracture history. All of the participants underwent a bone mineral density assessment by DXA, radiographs, and a serum C-peptide measurement. Thirty-four women presented fractures. Bivariate analysis revealed an inverse correlation between C-peptide and fractures (r = -0.27, p = 0.002). A significant difference in mean C-peptide levels was also found between women with vs. without fractures (p = 0.01, adjusted for age, BMI and glucose). Logistic regression analysis showed that C-peptide levels, femoral and vertebral BMD were all negatively associated with fracture status (B = -1.097, ES = 0.401, p = 0.006, 95% CI 0.15-0.73; B = -15.6, SE = 4.17, p C-peptide levels and a history of fractures in postmenopausal women without diabetes. These results suggest that C-peptidemay exert an effect on bone mineral density. However, further large-scale studies are needed to corroborate this finding and investigate the potential underlying mechanisms involved.

  10. Complete covalent structure of statherin, a tyrosine-rich acidic peptide which inhibits calcium phosphate precipitation from human parotid saliva.

    Science.gov (United States)

    Schlesinger, D H; Hay, D I

    1977-03-10

    The complete amino acid sequence of human salivary statherin, a peptide which strongly inhibits precipitation from supersaturated calcium phosphate solutions, and therefore stabilizes supersaturated saliva, has been determined. The NH2-terminal half of this Mr=5380 (43 amino acids) polypeptide was determined by automated Edman degradations (liquid phase) on native statherin. The peptide was digested separately with trypsin, chymotrypsin, and Staphylococcus aureus protease, and the resulting peptides were purified by gel filtration. Manual Edman degradations on purified peptide fragments yielded peptides that completed the amino acid sequence through the penultimate COOH-terminal residue. These analyses, together with carboxypeptidase digestion of native statherin and of peptide fragments of statherin, established the complete sequence of the molecule. The 2 serine residues (positions 2 and 3) in statherin were identified as phosphoserine. The amino acid sequence of human salivary statherin is striking in a number of ways. The NH2-terminal one-third is highly polar and includes three polar dipeptides: H2PO3-Ser-Ser-H2PO3-Arg-Arg-, and Glu-Glu-. The COOH-terminal two-thirds of the molecule is hydrophobic, containing several repeating dipeptides: four of -Gn-Pro-, three of -Tyr-Gln-, two of -Gly-Tyr-, two of-Gln-Tyr-, and two of the tetrapeptide sequence -Pro-Tyr-Gln-Pro-. Unusual cleavage sites in the statherin sequence obtained with chymotrypsin and S. aureus protease were also noted.

  11. Human C-peptide. Pt. 1

    International Nuclear Information System (INIS)

    Beischer, W.; Keller, L.; Maas, M.; Schiefer, E.; Pfeiffer, E.F.

    1976-01-01

    Synthetic human C-peptide bearing a tyrosine group at its amino end is labelled with 125 iodine using chloramin T or hydrogen peroxide and lactoperoxidase. The results of the two methods are compared. Antiserum to synthetic human C-peptide (without tyrosine), which was partially coupled to rabbit albumin, is raised in guinea pigs and goats. Goats show to be superior to guinea pips concerning antibody production. The so-called 'hook effect' phenomenon is observed when setting up the standard curves for the radioimmunoassay. Monotonically decreasing standard curves are obtained on dilution of antiserum with a high antibody titer which was produced by repeated immunization in goats. Free C-peptide and C-peptide bound to antiserum are separated using the anion exchange resin amberlite. Using this separation technique we excluded unspecific binding of labelled C-peptide to protein fractions in serum of diabetics. The sensitivity of our radioimmunoassay is approx. 0.3 ng C-peptide/ml serum. Intra- and interassay variability are below 10%. Human proinsulin is the only substance found to crossreact with the antiserum. (orig.) [de

  12. A study on the C-peptide radioimmunoassay with synthetized connecting peptide

    International Nuclear Information System (INIS)

    Nakagawa, Shoichi; Sasaki, Takashi; Nakayama, Hidetaka; Watanabe, Takuji; Aoki, Shin

    1976-01-01

    A method of C-peptide radioimmunoassay with the synthetized connecting peptide by Yanaihara was tested for the determination of serum C-peptide immunoreactivity (CPR) in normal people and in diabetics with or without insulin treatment. The CPR value obtained by this method was not interfered with by the presence of serum proteins or by the insulin of people with or without insulin treatment judged by the dilution test and the recovery test. The normal fasting CPR was 2.80 +- 0.78 ng/ml with the synthetized C-peptide as a standard. The CPR value increased and reached a maximum 90 minutes after the ingestion of 50 g of glucose. The increase after the glucose loading reduced corresponding to the severity of diabetes, and some juvenile-onset diabetes showed no response. Adult-type diabetics under insulin treatment, however, showed weak but significant CPR response. The increment of CPR and immunoreactive insulin after glucose loading in normal people and non-treated diabetics was well correlated (γ=0.8262). Judged from the above mentioned results, CPR determination in insulin-treated diabetics was thought to be a useful method for the assessment of the insulin-secreting ability of beta-cells of the pancreas. (J.P.N.)

  13. Connecting peptide (c-peptide) and the duration of diabetes mellitus ...

    African Journals Online (AJOL)

    Objective: C-peptide is derived from proinsulin and it is secreted in equimolar concentration with insulin. Plasma C-peptide is more stable than insulin and it provides an indirect measure of insulin secretory reserve and beta cell function. To determine relationship between C-peptide and duration of diabetes mellitus, age, ...

  14. The differences of saliva pH before and after application of Bis-GMA resin based-pit and fissure sealant

    Directory of Open Access Journals (Sweden)

    Ayub Irmadani Anwar

    2016-06-01

    Full Text Available Pit and fissure sealant applications is one way to prevent caries. One of the effective ingredient as a pit and fissure sealant is a Bis-GMA resin. One of the active components  of  Bis-GMA  resin  is  sodium  fluoride,  which is capable  of releasing fluoride ions to the tooth surface and able to improve the repair and remineralization process through the replacement  of  calcium and phosphate ions found in saliva to increase the pH of saliva. This study was to determine the effect of pit and fissure sealant applications made from Bis-GMA resin to pH of saliva in children aged 6-12 years. The study design is pre and post test with control group design. Saliva samples were derived from 15 pediatric patients has met the criteria. The pH of saliva were taken before and after the application of pit and fissure sealant made from Bis-GMA and measured with pH meter. This study used a non-parametric test, the Wilcoxon Signed Rank test. The results of this study are also supported by the results of statistical tests showed p-: 0.005 (p<0.05 value, which means that there are significant differences in pH of saliva before and after the application of pit and fissure sealant made from Bis-GMA resin.

  15. Intracellular Signalling by C-Peptide

    Directory of Open Access Journals (Sweden)

    Claire E. Hills

    2008-01-01

    Full Text Available C-peptide, a cleavage product of the proinsulin molecule, has long been regarded as biologically inert, serving merely as a surrogate marker for insulin release. Recent findings demonstrate both a physiological and protective role of C-peptide when administered to individuals with type I diabetes. Data indicate that C-peptide appears to bind in nanomolar concentrations to a cell surface receptor which is most likely to be G-protein coupled. Binding of C-peptide initiates multiple cellular effects, evoking a rise in intracellular calcium, increased PI-3-kinase activity, stimulation of the Na+/K+ ATPase, increased eNOS transcription, and activation of the MAPK signalling pathway. These cell signalling effects have been studied in multiple cell types from multiple tissues. Overall these observations raise the possibility that C-peptide may serve as a potential therapeutic agent for the treatment or prevention of long-term complications associated with diabetes.

  16. The influence of pilocarpine and biperiden on pH value and calcium, phosphate, and bicarbonate concentrations in saliva during and after radiotherapy for head and neck cancer.

    Science.gov (United States)

    Rode, M; smid, L; Budihna, M; Gassperssic, D; Rode, M; Soba, E

    2001-11-01

    The purpose of the present study was to investigate the influence of parasympathomimetic pilocarpine and anticholinergic biperiden on salivation, pH value, and calcium, phosphate, and bicarbonate concentrations in saliva in patients irradiated for malignant tumors of the head and neck region. Sixty-nine patients were randomly assigned into 3 groups. Group A consisted of patients receiving pilocarpine, group B of those who were receiving biperiden during radiotherapy and pilocarpine for 6 weeks after its completion, and group C comprised patients receiving neither of the mentioned drugs. The quantity of secreted unstimulated saliva, its pH value, as well as calcium, phosphate, and bicarbonate concentrations in saliva were measured before the beginning of radiotherapy, after 30 Gy of irradiation, at completed irradiation, and 3, 6, and 12 months after completion of radiotherapy. Saliva secretion was found to be the least affected in the group of patients receiving biperiden throughout the course of radiotherapy. One year after completion of therapy, the quantity of secreted saliva could only be measured in the patients receiving biperiden during radiotherapy; it amounted to 16% of the average initial quantity of saliva secreted before the beginning of irradiation. In all 3 groups of patients, mean pH value decreased during radiotherapy and started to increase again after completion of irradiation. In group B the decrease in pH value after radiotherapy was statistically significantly smaller than that in group C (P =.01). During and after irradiation, calcium concentration was increased in all 3 groups of patients. Phosphate concentration decreased during radiotherapy in all 3 groups. In group B it started to increase again 3 months after completion of radiotherapy. Bicarbonate concentration showed a slight increase during radiotherapy and started to decrease again after completion of irradiation. The results of our study indicate that the inhibition of saliva

  17. High-throughput screening of saliva for early detection of oral cancer: a pilot study.

    Science.gov (United States)

    Szanto, I; Mark, L; Bona, A; Maasz, G; Sandor, B; Gelencser, G; Turi, Z; Gallyas, F

    2012-04-01

    The success of tumour therapy depends considerably on early diagnosis. Therefore, we aimed to develop a widely available, cheap, non-invasive, high-throughput method suitable for screening high-risk populations, at least, for early signs of malignant transformation in the oral cavity. First, in order to identify suitable tumour marker candidates, we compared the protein patterns of five selected saliva samples obtained from healthy controls and tumour patients after electrophoretic separation, excised the bands that were consistently up-regulated in the tumour patients only, and performed matrix-assisted laser-desorption ionisation (MALDI)-time of flight (TOF) tandem mass spectrometry (MS/MS) analysis of the proteins in these bands after in-gel tryptic digestion. From the panel of proteins identified, we chose annexin 1 and peroxiredoxin 2 for further studies based on their presence in the saliva of all five oral cancer patients only. Then, we performed a homology search of protein databases using the primary sequence of each in silico tryptic fragment peptide of these two proteins as bait, and selected a unique peptide for each. Finally, we performed targeted MALDI-TOF MS peptide analysis in a blinded fashion on all samples obtained from 20 healthy controls and 22 tumour patients for the presence of these peptides. We found both peptides present in the saliva samples of all cancer patients only. Even though these tumour markers should be validated in a wider population, our results indicate that targeted MALDI-TOF MS analysis of unique peptides of putative saliva protein tumour biomarkers could be the method of choice for cost-efficient, high-throughput screening for the early detection of oral cancer.

  18. Family of pH-Low-Insertion-Peptides (pHLIPs)

    Science.gov (United States)

    Weerakkody, Dhammika; Moshnikova, Anna; Moshnikova, Valentina; Thakur, Mak; Rossi, Bethany; Engelman, Donald; Andreev, Oleg; Reshetnyak, Yana

    2012-02-01

    pHLIP (pH (Low) Insertion Peptide) is a novel delivery system for targeting of acidic diseased tissue such as solid tumors, sites of inflammation, arthritis and other pathological states. The molecular mechanism of pHLIP action is based on pH-dependent insertion and folding of pHLIP in membrane. We performed sequence variation and investigated 16 pHLIP variants with main goals of understanding the main principles of peptide-lipid interactions and tune delivery capability of pHLIP. The biophysical studies including thermodynamics and kinetics of the peptides interaction with a lipid bilayer of liposomes and cellular membranes were carried out. We found that peptides association to membrane at neutral and low pH could be modulated by 3-4 times. The apparent pK of transition from surface bound to membrane-inserted state could be tuned from 6.5 to 4.5. The rate of peptide's insertion across a bilayer could be enhanced 100 times compared to parent pHLIP. As a result, blood clearance and tumor targeting were modulated in a significant degree. The work is supported by NIH grants CA133890 to OAA, DME, YRK.

  19. Peptides for functionalization of InP semiconductors.

    Science.gov (United States)

    Estephan, Elias; Saab, Marie-belle; Larroque, Christian; Martin, Marta; Olsson, Fredrik; Lourdudoss, Sebastian; Gergely, Csilla

    2009-09-15

    The challenge is to achieve high specificity in molecular sensing by proper functionalization of micro/nano-structured semiconductors by peptides that reveal specific recognition for these structures. Here we report on surface modification of the InP semiconductors by adhesion peptides produced by the phage display technique. An M13 bacteriophage library has been used to screen 10(10) different peptides against the InP(001) and the InP(111) surfaces to finally isolate specific peptides for each orientation of the InP. MALDI-TOF/TOF mass spectrometry has been employed to study real affinity of the peptide towards the InP surfaces. The peptides serve for controlled placement of biotin onto InP to bind then streptavidin. Our Atomic Force Microscopy study revealed a total surface coverage of molecules when the InP surface was functionalized by its specific biotinylated peptide (YAIKGPSHFRPS). Finally, fluorescence microscopy has been employed to demonstrate the preferential attachment of the peptide onto a micro-patterned InP surface. Use of substrate specific peptides could present an alternative solution for the problems encountered in the actually existing sensing methods and molecular self-assembly due to the unwanted unspecific interactions.

  20. Production of peptide antisera specific for mouse and rat proinsulin C-peptide 2

    DEFF Research Database (Denmark)

    Blume, N; Madsen, O D; Kofod, Hans

    1990-01-01

    for antibody binding to the immunizing antigen. Antisera to C-peptide 2, stained islet beta-cells on mouse and rat, but not monkey pancreas sections in immunocytochemical analysis. Preabsorption to the synthetic C-peptide 2, but not the synthetic mouse and rat C-peptide 1 abolished staining. In conclusion we......Mice and rats have two functional non-allelic insulin genes. By using a synthetic peptide representing a common sequence in mouse and rat C-peptide 2 as antigen, we have produced rabbit antisera specific for an epitope which is not present in mouse or rat C-peptide 1. Long-term immunization did...... not seem to increase the end point titre as tested in direct ELISA. The specificity of the antiserum was determined by competitive ELISA and histochemistry on pancreas sections. Only the synthetic C-peptide 2, but not the homologous synthetic C-peptide 1 from mouse and rat competed efficiently in ELISA...

  1. Human C-peptide. Pt. 1. Radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Beischer, W; Keller, L; Maas, M; Schiefer, E; Pfeiffer, E F [Ulm Univ. (Germany, F.R.). Abt. Innere Medizin, Endokrinologie und Stoffwechsel

    1976-08-01

    Synthetic human C-peptide bearing a tyrosine group at its amino end is labelled with /sup 125/iodine using chloramin T or hydrogen peroxide and lactoperoxidase. The results of the two methods are compared. Antiserum to synthetic human C-peptide (without tyrosine), which was partially coupled to rabbit albumin, is raised in guinea pigs and goats. Goats show to be superior to guinea pips concerning antibody production. The so-called 'hook effect' phenomenon is observed when setting up the standard curves for the radioimmunoassay. Monotonically decreasing standard curves are obtained on dilution of antiserum with a high antibody titer which was produced by repeated immunization in goats. Free C-peptide and C-peptide bound to antiserum are separated using the anion exchange resin amberlite. Using this separation technique we excluded unspecific binding of labelled C-peptide to protein fractions in serum of diabetics. The sensitivity of our radioimmunoassay is approx. 0.3 ng C-peptide/ml serum. Intra- and interassay variability are below 10%. Human proinsulin is the only substance found to crossreact with the antiserum.

  2. Basal C-peptide Level as a Surrogate Marker of Subclinical Atherosclerosis in Type 2 Diabetic Patients

    Directory of Open Access Journals (Sweden)

    Sung-Tae Kim

    2011-02-01

    Full Text Available BackgroundRecent studies have revealed that C-peptide induces smooth muscle cell proliferation and causes human atherosclerotic lesions in diabetic patients. The present study was designed to examine whether the basal C-peptide levels correlate with cardiovascular risk in type 2 diabetes mellitus (T2DM patients.MethodsData was obtained from 467 patients with T2DM from two institutions who were followed for four years. The medical findings of all patients were reviewed, and patients with creatinine >1.4 mg/dL, any inflammation or infection, hepatitis, or type 1 DM were excluded. The relationships between basal C-peptide and other clinical values were statistically analyzed.ResultsA simple correlation was found between basal C-peptide and components of metabolic syndrome (MS. Statistically basal C-peptide levels were significantly higher than the three different MS criteria used in the present study, the Adult Treatment Panel III (ATP III of the National Cholesterol Education Program's (NCEP's, World Health Organization (WHO, and the International Diabetes Federation (IDF criteria (NCEP-ATP III, P=0.001; IDF, P<0.001; WHO, P=0.029. The multiple regression analysis between intima-media thickness (IMT and clinical values showed that basal C-peptide significantly correlated with IMT (P=0.043, while the analysis between the 10-year coronary heart disease risk by the United Kingdom Prospective Diabetes Study risk engine and clinical values showed that basal C-peptide did not correlate with IMT (P=0.226.ConclusionBasal C-peptide is related to cardiovascular predictors (IMT of T2DM, suggesting that basal C-peptide does provide a further indication of cardiovascular disease.

  3. Stability and reproducibility of a single-sample urinary C-peptide/creatinine ratio and its correlation with 24-h urinary C-peptide.

    Science.gov (United States)

    McDonald, Tim J; Knight, Bridget A; Shields, Beverley M; Bowman, Pamela; Salzmann, Maurice B; Hattersley, Andrew T

    2009-11-01

    C-peptide measurement in blood or 24-h urine samples provides useful information regarding endogenous insulin secretion, but problems related to the rapid degradation of C-peptide in blood and difficulty of 24-h urine collection have limited widespread routine clinical use of this test. We assessed the feasibility of measuring urinary C-peptide (UCP) with correction for creatinine concentration in single urine samples. We analyzed UCP using a routine electrochemiluminescence immunoassay in samples from 21 healthy volunteers. We investigated the stability of UCP with different preservatives and storage conditions and compared the reproducibility of urinary C-peptide/creatinine ratio (UCPCR) in first- and second-void fasting urines, then assessed correlations with 24-h collections. UCPCR was unchanged at room temperature for 24 h and at 4 degrees C for 72 h even in the absence of preservative. UCPCR collected in boric acid was stable at room temperature for 72 h. UCPCR remained stable after 7 freeze-thaw cycles but decreased with freezer storage time and dropped to 82%-84% of baseline by 90 days at -20 degrees C. Second-void fasting UCPCRs were lower than first-void (median 0.78 vs 1.31, P = 0.0003) and showed less variation (CV 33% vs 52%), as second-void UCPCRs were not influenced by evening food-related insulin secretion. Second-void fasting UCPCR was highly correlated with 24-h UCP (r = 0.8, P = 0.00006). Second-void fasting UCPCR is a reproducible measure that correlates well with 24-h UCP in normal samples. The 3-day stability of UCPCR at room temperature greatly increases its potential clinical utility.

  4.  Pleiotropic action of proinsulin C-peptid

    Directory of Open Access Journals (Sweden)

    Michał Usarek

    2012-03-01

    Full Text Available  Proinsulin C-peptide, released in equimolar amounts with insulin by pancreatic β cells, since its discovery in 1967 has been thought to be devoid of biological functions apart from correct insulin processing and formation of disulfide bonds between A and B chains. However, in the last two decades research has brought a substantial amount of data indicating a crucial role of C-peptide in regulating various processes in different types of cells and organs. C-peptide acts presumably via either G-protein-coupled receptor or directly inside the cell, after being internalized. However, a receptor binding this peptide has not been identified yet. This peptide ameliorates pathological changes induced by type 1 diabetes mellitus, including glomerular hyperfiltration, vessel endothelium inflammation and neuron demyelinization. In diabetic patients and diabetic animal models, C-peptide substitution in physiological doses improves the functional and structural properties of peripheral neurons and protects against hyperglycemia-induced apoptosis, promoting neuronal development, regeneration and cell survival. Moreover, it affects glycogen synthesis in skeletal muscles. In vitro C-peptide promotes disaggregation of insulin oligomers, thus enhancing its bioavailability and effects on metabolism. There are controversies concerning the biological action of C-peptide, particularly with respect to its effect on Na /K -ATPase activity. Surprisingly, the excess of circulating peptide associated with diabetes type 2 contributes to atherosclerosis development. In view of these observations, long-term, large-scale clinical investigations using C-peptide physiological doses need to be conducted in order to determine safety and health outcomes of long-term administration of C-peptide to diabetic patients.

  5. Novel thrombopoietin mimetic peptides bind c-Mpl receptor: Synthesis, biological evaluation and molecular modeling.

    Science.gov (United States)

    Liu, Yaquan; Tian, Fang; Zhi, Dejuan; Wang, Haiqing; Zhao, Chunyan; Li, Hongyu

    2017-02-01

    Thrombopoietin (TPO) acts in promoting the proliferation of hematopoietic stem cells and by initiating specific maturation events in megakaryocytes. Now, TPO-mimetic peptides with amino acid sequences unrelated to TPO are of considerable pharmaceutical interest. In the present paper, four new TPO mimetic peptides that bind and activate c-Mpl receptor have been identified, synthesized and tested by Dual-Luciferase reporter gene assay for biological activities. The molecular modeling research was also approached to understand key molecular mechanisms and structural features responsible for peptide binding with c-Mpl receptor. The results presented that three of four mimetic peptides showed significant activities. In addition, the molecular modeling approaches proved hydrophobic interactions were the driven positive forces for binding behavior between peptides and c-Mpl receptor. TPO peptide residues in P7, P13 and P7' positions were identified by the analysis of hydrogen bonds and energy decompositions as the key ones for benefiting better biological activities. Our data suggested the synthesized peptides have considerable potential for the future development of stable and highly active TPO mimetic peptides. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Diagnostic value of C-peptide determination

    International Nuclear Information System (INIS)

    Kober, G.; Rainer, O.H.

    1983-01-01

    C-peptide and insulin serum determinations were performed in 94 glucagon-stimulated diabetics and in 15 healthy persons. A minimal increase of 1.5 ng C-peptide/ml serum after glucagon injection (1 mg i.v.) was found to be a useful parameter for the differentiation of insulin dependent and non-insulin dependent diabetics. The maximal response to glucagon occurred during the first 10-minutes after the injection (blood was drawn at 2-minutes intervals). Serum insulin levels and basal C-peptide concentrations were of no value in predicting insulin-dependency. Basal C-peptide levels were significantly different from control in juvenile insulin dependent diabetics (decrease) only. (Author)

  7. Erosion protection conferred by whole human saliva, dialysed saliva, and artificial saliva

    Science.gov (United States)

    Baumann, T.; Kozik, J.; Lussi, A.; Carvalho, T. S.

    2016-10-01

    During dental erosion, tooth minerals are dissolved, leading to a softening of the surface and consequently to irreversible surface loss. Components from human saliva form a pellicle on the tooth surface, providing some protection against erosion. To assess the effect of different components and compositions of saliva on the protective potential of the pellicle against enamel erosion, we prepared four different kinds of saliva: human whole stimulated saliva (HS), artificial saliva containing only ions (AS), human saliva dialysed against artificial saliva, containing salivary proteins and ions (HS/AS), and human saliva dialysed against deionised water, containing only salivary proteins but no ions (HS/DW). Enamel specimens underwent four cycles of immersion in either HS, AS, HS/AS, HS/DW, or a humid chamber (Ctrl), followed by erosion with citric acid. During the cycling process, the surface hardness and the calcium released from the surface of the specimens were measured. The different kinds of saliva provided different levels of protection, HS/DW exhibiting significantly better protection than all the other groups (p < 0.0001). Different components of saliva, therefore, have different effects on the protective properties of the pellicle and the right proportions of these components in saliva are critical for the ability to form a protective pellicle.

  8. Fall in C-Peptide During First 2 Years From Diagnosis

    Science.gov (United States)

    Greenbaum, Carla J.; Beam, Craig A.; Boulware, David; Gitelman, Stephen E.; Gottlieb, Peter A.; Herold, Kevan C.; Lachin, John M.; McGee, Paula; Palmer, Jerry P.; Pescovitz, Mark D.; Krause-Steinrauf, Heidi; Skyler, Jay S.; Sosenko, Jay M.

    2012-01-01

    Interpretation of clinical trials to alter the decline in β-cell function after diagnosis of type 1 diabetes depends on a robust understanding of the natural history of disease. Combining data from the Type 1 Diabetes TrialNet studies, we describe the natural history of β-cell function from shortly after diagnosis through 2 years post study randomization, assess the degree of variability between patients, and investigate factors that may be related to C-peptide preservation or loss. We found that 93% of individuals have detectable C-peptide 2 years from diagnosis. In 11% of subjects, there was no significant fall from baseline by 2 years. There was a biphasic decline in C-peptide; the C-peptide slope was −0.0245 pmol/mL/month (95% CI −0.0271 to −0.0215) through the first 12 months and −0.0079 (−0.0113 to −0.0050) from 12 to 24 months (P < 0.001). This pattern of fall in C-peptide over time has implications for understanding trial results in which effects of therapy are most pronounced early and raises the possibility that there are time-dependent differences in pathophysiology. The robust data on the C-peptide obtained under clinical trial conditions should be used in planning and interpretation of clinical trials. PMID:22688329

  9. Insulin and C-peptide in human brain neurons (insulin/C-peptide/brain peptides/immunohistochemistry/radioimmunoassay)

    International Nuclear Information System (INIS)

    Dorn, A.; Bernstein, H.G.; Rinne, A.; Hahn, H.J.; Ziegler, M.

    1983-01-01

    The regional distribution and cellular localization of insulin and C-peptide immunoreactivities were studied in human cadaver brains using the indirect immunofluorescence method, the peroxidase-antiperoxidase technique, and radioimmunoassay. Products of the immune reactions to both polypeptides were observed in most nerve cells in all areas of the brain examined. Immunostaining was mainly restricted to the cell soma and proximal dendrites. Radioimmunoassay revealed that human brain contains insulin and C-peptide in concentrations much higher than the blood, the highest being in the hypothalamus. These findings support the hypothesis that the 'brain insulin' is - at least in part - produced in the CNS. (author)

  10. An Optical Sensor with Polyaniline-Gold Hybrid Nanostructures for Monitoring pH in Saliva

    Directory of Open Access Journals (Sweden)

    Chongdai Luo

    2017-03-01

    Full Text Available Saliva contains important personal physiological information that is related to some diseases, and it is a valuable source of biochemical information that can be collected rapidly, frequently, and without stress. In this article, we reported a new and simple localized surface plasmon resonance (LSPR substrate composed of polyaniline (PANI-gold hybrid nanostructures as an optical sensor for monitoring the pH of saliva samples. The overall appearance and topography of the substrates, the composition, and the wettability of the LSPR surfaces were characterized by optical and scanning electron microscope (SEM images, infrared spectra, and contact angles measurement, respectively. The PANI-gold hybrid substrate readily responded to the pH. The response time was very short, which was 3.5 s when the pH switched from 2 to 7, and 4.5 s from 7 to 2. The changes of visible-near-infrared (NIR spectra of this sensor upon varying pH in solution showed that—for the absorption at given wavelengths of 665 nm and 785 nm—the sensitivities were 0.0299 a.u./pH (a.u. = arbitrary unit with a linear range of pH = 5–8 and 0.0234 a.u./pH with linear range of pH = 2–8, respectively. By using this new sensor, the pH of a real saliva sample was monitored and was consistent with the parallel measurements with a standard laboratory method. The results suggest that this novel LSPR sensor shows great potential in the field of mobile healthcare and home medical devices, and could also be modified by different sensitive materials to detect various molecules or ions in the future.

  11. An Optical Sensor with Polyaniline-Gold Hybrid Nanostructures for Monitoring pH in Saliva.

    Science.gov (United States)

    Luo, Chongdai; Wang, Yangyang; Li, Xuemeng; Jiang, Xueqin; Gao, Panpan; Sun, Kang; Zhou, Jianhua; Zhang, Zhiguang; Jiang, Qing

    2017-03-17

    Saliva contains important personal physiological information that is related to some diseases, and it is a valuable source of biochemical information that can be collected rapidly, frequently, and without stress. In this article, we reported a new and simple localized surface plasmon resonance (LSPR) substrate composed of polyaniline (PANI)-gold hybrid nanostructures as an optical sensor for monitoring the pH of saliva samples. The overall appearance and topography of the substrates, the composition, and the wettability of the LSPR surfaces were characterized by optical and scanning electron microscope (SEM) images, infrared spectra, and contact angles measurement, respectively. The PANI-gold hybrid substrate readily responded to the pH. The response time was very short, which was 3.5 s when the pH switched from 2 to 7, and 4.5 s from 7 to 2. The changes of visible-near-infrared (NIR) spectra of this sensor upon varying pH in solution showed that-for the absorption at given wavelengths of 665 nm and 785 nm-the sensitivities were 0.0299 a.u./pH (a.u. = arbitrary unit) with a linear range of pH = 5-8 and 0.0234 a.u./pH with linear range of pH = 2-8, respectively. By using this new sensor, the pH of a real saliva sample was monitored and was consistent with the parallel measurements with a standard laboratory method. The results suggest that this novel LSPR sensor shows great potential in the field of mobile healthcare and home medical devices, and could also be modified by different sensitive materials to detect various molecules or ions in the future.

  12. Saliva-catalyzed hydrolysis of a ketobemidone ester prodrug

    DEFF Research Database (Denmark)

    Hansen, L.B.; Christrup, Lona Louring; Bundgaard, H.

    1992-01-01

    Saliva enzyme-catalysed hydrolysis of ester prodrugs or drugs containing sensitive ester groups may be a limiting factor for the buccal absorption of such compounds. Using the isopropyl carbonate ester of ketobemidone as a model substance of a hydrolysis-sensitive prodrug the esterase activity...... of human saliva has been characterized as a function of various factors. The esterase activity was found to decrease rapidly upon storage of the saliva at 37°C. The activity increased with increasing pH in the range 4.5-7.4 and with increasing salivation flow rate up to a rate of 0.9 ml min. Under resting...... conditions, the flow rate was about 0.2 ml min which implied a greatly decreased esterase activity. The activity was highest after fasting and decreased after intake of a meal. The intraindividual variation in the saliva esterase activity was small whereas a larger interindividual variation was found....

  13. Lassomycin, a ribosomally synthesized cyclic peptide, kills mycobacterium tuberculosis by targeting the ATP-dependent protease ClpC1P1P2.

    Science.gov (United States)

    Gavrish, Ekaterina; Sit, Clarissa S; Cao, Shugeng; Kandror, Olga; Spoering, Amy; Peoples, Aaron; Ling, Losee; Fetterman, Ashley; Hughes, Dallas; Bissell, Anthony; Torrey, Heather; Akopian, Tatos; Mueller, Andreas; Epstein, Slava; Goldberg, Alfred; Clardy, Jon; Lewis, Kim

    2014-04-24

    Languishing antibiotic discovery and flourishing antibiotic resistance have prompted the development of alternative untapped sources for antibiotic discovery, including previously uncultured bacteria. Here, we screen extracts from uncultured species against Mycobacterium tuberculosis and identify lassomycin, an antibiotic that exhibits potent bactericidal activity against both growing and dormant mycobacteria, including drug-resistant forms of M. tuberculosis, but little activity against other bacteria or mammalian cells. Lassomycin is a highly basic, ribosomally encoded cyclic peptide with an unusual structural fold that only partially resembles that of other lasso peptides. We show that lassomycin binds to a highly acidic region of the ClpC1 ATPase complex and markedly stimulates its ATPase activity without stimulating ClpP1P2-catalyzed protein breakdown, which is essential for viability of mycobacteria. This mechanism, uncoupling ATPase from proteolytic activity, accounts for the bactericidal activity of lassomycin. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Rabbit IgG directed to a synthetic C-terminal peptide of the major grass pollen allergen Lol p I inhibits human basophil histamine release induced by natural Lol p I.

    Science.gov (United States)

    van Ree, R; Aalberse, R C

    1995-03-01

    The potential role of allergen-specific IgG antibodies as 'blocking' antibodies in allergen-induced human basophil histamine release was investigated. This was studied in a model with the major grass pollen allergen Lol p I and polyclonal rabbit antisera directed against this allergen and against a synthetic peptide of its C terminus. When allergen and antibodies were allowed to preincubate, Lol p I induced histamine release was inhibited up to 85% by the antiserum against Lol p I. By omitting preincubation, and thereby more closely mimicking an in vivo situation, up to 55% inhibition was realized. This indicates that allergen-specific IgG can act as 'blocking' antibody without preincubation. Immunization of rabbits with a synthetic C-terminal peptide of Lol p I resulted in antibodies reactive with natural Lol p I. Despite their 100-fold lower avidity for Lol p I (as compared with antinatural Lol p I), these antibodies had the capacity to inhibit Lol p I induced histamine release for > 90% (up to 50% without preincubation). This indicates that it is possible to block histamine release induced by a major allergen with low-avidity IgG antibodies directed against a minor proportion of the allergen (25 amino acids). IgE antibodies from the donors studied were unreactive with this synthetic peptide, indicating that for blocking activity identical epitope specificity of IgE and IgG is not essential. This opens interesting perspectives for application of synthetic peptides in immunotherapy, distinct from their effects on T cell reactivity.

  15. Concentration of total protein and degree of acidity (pH of saliva when fasting and after breakfasting

    Directory of Open Access Journals (Sweden)

    Gemella Nur Illahi

    2016-04-01

    Full Text Available Background: While fasting, the mouth does not work to eat and drink so that the salivary glands become less active so saliva production decreased and there was a change in eating timewhich is relation to the mastication process that impact on changes in the degree of acidity (pH Objectives: To determine the concentration of total protein and the degree of acidity (pH of saliva when fasting and after breakfasting. Materials and Methods: The study was observational analytic design with longitudinal (follow up study conducted in the Hj. Halima Dg. Sikati Dental Hospital inKandea in July 2015, the sampling method was purposive sampling. Population was 35 clinical students at the Department of Dental Public Health, Faculty of Dentistry Hasanuddin University with a total sample of 16 students who fit the criteria of the study subjects. To calculate the total protein of saliva concentration using Kyltecautoanalyzerand pH meter to measure the acidity of saliva. Data was analyzed was using SPSS version 17.0 (paired t-test, p <0.05. Results: The mean of total protein (% while fasting by 0135% ± 0.026 and the mean total protein (% after breakfasting at 0.179% ± 0.035, while the average degree of acidity (pH during fasting at 7.26 ± 0:24 and the average degree of acidity (pH after breakfasting at 7.66 ± 0.23 with p-value (0.000. Conclusions: An increase in the total protein concentration and acidity (pH after breakfasting.

  16. Effect of Mobile Phone Usage on Nickel Ions Release and pH of Saliva in Patients Undergoing Fixed Orthodontic Treatment.

    Science.gov (United States)

    Nanjannawar, Lalita Girish; Girme, Tejashree Suresh; Agrawal, Jiwanasha Manish; Agrawal, Manish Suresh; Fulari, Sangamesh Gurunath; Shetti, Shraddha Subhash; Kagi, Vishwal Ajith

    2017-09-01

    Hand held mobile phones are presently the most popular means of communication worldwide and have transformed our lives in many aspects. The widespread use of such devices have resulted in growing concerns regarding harmful effects of radiations emitted by them. This study was designed to evaluate the effects of mobile phone usage on nickel ion release as well as pH of saliva in patients with fixed orthodontic appliances. To assess the level of nickel ions in saliva and pH of saliva in mobile phone users undergoing fixed orthodontic treatment using inductively coupled plasma atomic emission spectrometry. A total of 42 healthy patients with fixed orthodontic appliance in mouth for a duration of six to nine months were selected for the study. They were divided into experimental group (n=21) consisting of mobile phone users and control group (n=21) of non mobile phone users. Saliva samples were collected from both the groups and nickel ion levels were measured using inductively coupled plasma-mass spectroscopy. The pH values were also assessed for both groups using pH meter. Unpaired t-test was used for the data analysis. Statistical analysis revealed that though the pH levels were reduced and the nickel ion levels were higher in the experimental group compared to the control group, the results were non significant. Mobile phone usage may affect the pH of saliva and result in increased release of nickel ions in saliva of patients with fixed orthodontic appliances in the oral cavity.

  17. Constructing bioactive peptides with pH-dependent activities.

    Science.gov (United States)

    Tu, Zhigang; Volk, Melanie; Shah, Khushali; Clerkin, Kevin; Liang, Jun F

    2009-08-01

    Many bioactive peptides are featured by their arginine and lysine rich contents. In this study, lysine and arginine residues in lytic peptides were selectively replaced by histidines. Although resulting histidine-containing lytic peptides had decreased activity, they did show pH-dependent cytotoxicity. The activity of the constructed histidine-containing lytic peptides increased 2-8 times as the solution pH changed from 7.4 to 5.5. More importantly, these histidine-containing peptides maintain the same cell killing mechanism as their parent peptides by causing cell lysis. Both the activity and pH-sensitivity of histidine-containing peptides are tunable by adjusting histidine substitution numbers and positions. This study has presented a general strategy to create bioactive peptides with desired pH-sensitivity to meet the needs of various applications such as cancer treatments.

  18. Protective effect of C-peptide on experimentally induced diabetic nephropathy and the possible link between C-peptide and nitric oxide.

    Science.gov (United States)

    Elbassuoni, Eman A; Aziz, Neven M; El-Tahawy, Nashwa F

    2018-06-01

    Diabetic nephropathy one of the major microvascular diabetic complications. Besides hyperglycemia, other factors contribute to the development of diabetic complications as the proinsulin connecting peptide, C-peptide. We described the role of C-peptide replacement therapy on experimentally induced diabetic nephropathy, and its potential mechanisms of action by studying the role of nitric oxide (NO) as a mediator of C-peptide effects by in vivo modulating its production by N G -nitro-l-arginine methyl ester (L-NAME). Renal injury markers measured were serum urea, creatinine, tumor necrosis factor alpha, and angiotensin II, and malondialdehyde, total antioxidant, Bcl-2, and NO in renal tissue. In conclusion, diabetic induction resulted in islet degenerations and decreased insulin secretion with its metabolic consequences and subsequent renal complications. C-Peptide deficiencies in diabetes might have contributed to the metabolic and renal error, since C-peptide treatment to the diabetic rats completely corrected these errors. The beneficial effects of C-peptide are partially antagonized by L-NAME coadministration, indicating that NO partially mediates C-peptide effects.

  19. Usefulness of saliva for measurement of 3,4-methylenedioxymethamphetamine and its metabolites: correlation with plasma drug concentrations and effect of salivary pH.

    Science.gov (United States)

    Navarro, M; Pichini, S; Farré, M; Ortuño, J; Roset, P N; Segura, J; de la Torre, R

    2001-10-01

    Saliva is an alternative biologic matrix for drugs-of-abuse testing that offers the advantages of noninvasive, rapid, and easy sampling. We studied the excretion profile of 3,4-methylenedioxymethamphetamine (MDMA) and its metabolites in both saliva and plasma, as well the effect of the drug on salivary pH. Saliva and plasma samples were obtained from eight healthy MDMA consumers after ingestion of a single 100-mg dose of the drug. Concentrations of MDMA and its main metabolites, 3,4-methylenedioxyamphetamine (MDA) and 4-hydroxy-3-methoxymethamphetamine (HMMA), in saliva and plasma were measured by gas chromatography-mass spectrometry. Apparent pharmacokinetic parameters for MDMA in saliva were estimated, and the saliva-to-plasma ratio at each time interval was calculated and correlated with salivary pH. MDMA, MDA, and HMMA were detected in saliva. Salivary concentrations of MDMA were 1728.9-6510.6 microg/L and peaked at 1.5 h after drug intake. This was followed by a progressive decrease, with a mean concentration of 126.2 microg/L at 24 h. The saliva-to-plasma ratio was 32.3-1.2, with a peak of 18.1 at 1.5 h after drug administration. Salivary pH seemed to be affected by MDMA administration; pH values decreased by 0.6 units (mean pH values of 6.9 and 6.8 at 1.5 and 4 h after drug administration vs predose pH of 7.4). Measurement of MDMA in saliva is a valuable alternative to determination of plasma drug concentrations in both clinical and toxicologic studies. On-site testing is also facilitated by noninvasive and rapid collection of salivary specimens.

  20. Influence of C-Peptide on Glucose Utilisation

    Directory of Open Access Journals (Sweden)

    B. Wilhelm

    2008-01-01

    Full Text Available During the recent years, multiple studies demonstrated that C-peptide is not an inert peptide, but exerts important physiological effects. C-peptide binds to cell membranes, stimulates the Na,K-ATPase and the endothelial nitric oxide (NO synthase. Moreover, there is evidence that C-peptide decreases glomerular hyperfiltration and increases glucose utilisation. Nevertheless, there is still limited knowledge concerning mechanisms leading to an increased glucose utilisation either in rats or in humans. The aim of this paper is to give an overview over the published studies regarding C-peptide and glucose metabolism from in vitro studies to longer lasting studies in humans.

  1. Does bombesin-like peptide mediate radiation-induced anorexia and satiety?

    Energy Technology Data Exchange (ETDEWEB)

    Aalto, Y.; Franzen, L.; Henriksson, R. [Umeaa Univ. (Sweden). Dept. of Oncology; Forsgren, S.; Kjoerell, U. [Umeaa Univ. (Sweden). Dept. of Anatomy; Funegaard, U. [Umeaa Univ. (Sweden). Dept. of Cardiology

    1999-07-01

    Bombesin (BN) and its mammalian counterpart gastrin-releasing peptide (GRP) act as neuroregulatory hormones and peripheral and central satiety-inducing agents. Previously, we demonstrated that irradiation induces an increase in the expression of BN/GRP in the innervation of the salivary glands in rats. We therefore carried out a study using radioimmunoassay (RIA) analysis and immunohistochemistry to examine whether saliva contains BN and whether irradiation affects the BN release to saliva in rats. Immunoreactivity for BN was detected not only in the innervation of the parenchyma but also in the duct cells and in the lumina of the ducts, suggesting entrance of BN into saliva. The RIA analysis confirmed that rat saliva contains a BN-like peptide. The observation shows that saliva contains this peptide but that there is no significant increase following the radiation schedule used. Nevertheless, the occurrence of an enhanced expression of BN in different peripheral tissues such as the salivary and laryngeal glands should be taken into consideration when discussing the clinically important problem of reduced food intake and anorexia in cancer patients. (orig.)

  2. Does bombesin-like peptide mediate radiation-induced anorexia and satiety?

    International Nuclear Information System (INIS)

    Aalto, Y.; Franzen, L.; Henriksson, R.; Forsgren, S.; Kjoerell, U.; Funegaard, U.

    1999-01-01

    Bombesin (BN) and its mammalian counterpart gastrin-releasing peptide (GRP) act as neuroregulatory hormones and peripheral and central satiety-inducing agents. Previously, we demonstrated that irradiation induces an increase in the expression of BN/GRP in the innervation of the salivary glands in rats. We therefore carried out a study using radioimmunoassay (RIA) analysis and immunohistochemistry to examine whether saliva contains BN and whether irradiation affects the BN release to saliva in rats. Immunoreactivity for BN was detected not only in the innervation of the parenchyma but also in the duct cells and in the lumina of the ducts, suggesting entrance of BN into saliva. The RIA analysis confirmed that rat saliva contains a BN-like peptide. The observation shows that saliva contains this peptide but that there is no significant increase following the radiation schedule used. Nevertheless, the occurrence of an enhanced expression of BN in different peripheral tissues such as the salivary and laryngeal glands should be taken into consideration when discussing the clinically important problem of reduced food intake and anorexia in cancer patients. (orig.)

  3. Saliva C-reactive protein as a biomarker of metabolic syndrome in diabetic patients.

    Science.gov (United States)

    Dezayee, Zhian Mahmood Ibrahim; Al-Nimer, Marwan Salih Mohamad

    2016-01-01

    Human C-reactive protein (CRP) has been used in the risk assessment of coronary events. Human saliva mirrors the body's health and well-being and is noninvasive, easy to collect, and ideal for third-world countries as well as for large patient screening. This study aimed to screen the saliva CRP qualitatively in patients with diabetes (Type 1 and 2) taking in considerations, the diagnostic criteria of metabolic syndrome. Center for diabetes mellitus, prospective study. A total number of 50 Type 2 diabetes (T2D) patients, 25 Type 1 diabetes (T1D) patients, and 25 healthy subjects were recruited from the center for diabetes mellitus. Each patient was assessed clinically, and the anthropometric measures, glycemic status, and lipid profiles were determined. Stimulated salivary flow rate and saliva CRP were determined. All calculations analysis was made using Excel 2003 program for Windows. The results showed that the salivary flow rate in T1D was less than healthy subjects and T2D and CRP was found positive (6 mg/L) in 36% and 56% of patients with T1D and T2D, respectively. Saliva CRP was found to be related to the anthropometric measurement, blood pressure, and glycemic control. We conclude that saliva CRP may be used as a biomarker for metabolic syndrome and its value is obvious in T2D rather than in T1D.

  4. Cigarette smoke-exposed saliva suppresses cellular and humoral immune responses in an animal model

    International Nuclear Information System (INIS)

    Jafarzadeh, A.; Bakhshi, H.; Rezayati, M.T.; Nemati, M.

    2009-01-01

    To evaluate the effects of cigarette smoke (CS)-exposed saliva on cellular and antibody responses in an animal model. The stimulatory and non-stimulatory saliva samples were collected from 10 healthy subjects and were then exposed to CS for 20 or 80 minutes. The CS-exposed saliva samples were administrated intraperitoneally (i.p) to male Balb/c mice. Then the delayed type hypersensitivity (DTH) and antibody responses to sheep red blood cell (SRBC) was assessed. Moreover, the total white blood cells (WBC) counts and the blood lymphocytes counts were determined. The mean of DTH responses of animal groups received 20 minutes or 80 minutes CS-exposed saliva samples was significantly lower than that observed in control group. Moreover, The mean titer of anti-SRBC antibody was significantly lower in animal groups who received 80 minutes CS-exposed stimulatory or non-stimulatory saliva as compared to control group (P<0.04 and P<0.002, respectively). The mean counts of blood lymphocytes in 80 minutes CS exposed-stimulatory saliva group was also significantly lower as compared to control group (P<0.05). These results show that the CS-exposed saliva samples have profound suppressive effects on both cellular and humoral immune response in a mouse animal model (JPMA 59:760; 2009). (author)

  5. Peptide Fraction pOh2 Exerts Antiadipogenic Activity through Inhibition of C/EBP-α and PPAR-γ Expression in 3T3-L1 Adipocytes

    Directory of Open Access Journals (Sweden)

    Thi Tuyet Nhung Nguyen

    2017-01-01

    Full Text Available Many studies have comprehensively examined the venom of Ophiophagus hannah snake. Its venom comprises different compounds exhibiting a wide range of pharmacological activities. In this investigation, four peptide fractions (PFs, ranging from 3 kDa to 10 kDa, isolated from the Vietnamese snake venom of O. hannah were separated by HPLC and investigated for their inhibitory activity on adipogenesis in 3T3-L1 adipocytes. The most effective PF was then further purified, generating two peptides, pOh1 and pOh2. Upon investigation of these two peptides on 3T3-L1 adipocytes, it was revealed that, at 10 μg/mL, pOh2 was able to inhibit the lipid accumulation in 3T3-L1 adipocytes by up to 56%, without affecting cell viability. Furthermore, the pOh2 downregulated the gene expression of important transcription factors C/EBP-α and PPAR-γ. In addition, aP2 and GPDH adipocyte-specific markers were also significantly reduced compared to untreated differentiated cells. Taken together, pOh2 inhibited the expression of key transcription factors C/EBP-α and PPAR-γ and their target genes, aP2 and GPDH, thereby blocking the adipocyte differentiation. In conclusion, this novel class of peptide might have potential for in vivo antiobesity effects.

  6. Novel tetra-peptide insertion in Gag-p6 ALIX-binding motif in HIV-1 subtype C associated with protease inhibitor failure

    Science.gov (United States)

    Neogi, Ujjwal; RAO, Shwetha D; BONTELL, Irene; VERHEYEN, Jens; RAO, Vasudev R; GORE, Sagar C; SONI, Neelesh; SHET, Anita; SCHÜLTER, Eugen; EKSTRAND, Maria L.; WONDWOSSEN, Amogne; KAISER, Rolf; MADHUSUDHAN, Mallur S.; PRASAD, Vinayaka R; SONNERBORG, Anders

    2014-01-01

    A novel tetra-peptide insertion was identified in Gag-p6 ALIX-binding region which is appears in protease inhibitor (PI) failure Indian HIV-1C sequences (Odds Ratio 17.1, p<0.001) but naturally present in half of untreated Ethiopian sequences. The insertion will probably restore the ALIX mediated virus release pathway, which is lacking in HIV-1C. The clinical importance of such insertion need to be evaluated in HIV-1C dominating regions were PI-drugs are being scaled up as second line treatment options. PMID:25102091

  7. The relationship between C-type natriuretic peptide and cognitive impairment in older patients with Type 2 diabetes

    International Nuclear Information System (INIS)

    Li Xinling; Zhu Xiangyang; Huang Huaiyu; Jin Yan

    2011-01-01

    Objective: To investigate the relationship between C-type natriuretic peptide and cognitive impairment in older patients with type 2 diabetes, and to explore the pathogenesis of diabetic cognitive impairment. Methods: According to the Montreal Cognitive Assessment (MoCA) scores, 80 type 2 diabetic patients over the age of 60 years were divided into two groups, one group including 31 cases with cognitive impairment, the other 49 patients with non-cognitive impairment. And 80 normal participants were selected as the control group. Plasma level of C-type natriuretic peptide was measured by radio-immunity assay in all subjects. The changes and associations of the plasma C-type natriuretic peptide level among three groups was analyzed. Result: In the non-cognitive impairment group, plasma level of C-type natriuretic peptide was higher than that in the control group (P<0.01). But the plasma level of C-type natriuretic peptide in the cognitive impairment group was degraded, significantly deferent with those in the control group and the non-cognitive impairment group (P<0.01). MoCA scores of the cognitive impairment group positively correlated with plasma level of C-type natriuretic peptide (r=0.513, P<0.01). Conclusion: In the early period of type 2 diabetes,the secretion of C-type natriuretic peptide was increased. When diabetic cognitive impairment complicated,the secretion of C-type natriuretic peptide was decompensated. Then plasma level of C-type natriuretic peptide become low. The level of C-type natriuretic peptide closely correlated with diabetic cognitive impairment. It was suggested that diabetic angiopathies may act an important role in the pathogenesis of diabetic cognitive impairment. (authors)

  8. A dietary pattern that is associated with C-peptide and risk of colorectal cancer in women.

    Science.gov (United States)

    Fung, Teresa T; Hu, Frank B; Schulze, Matthias; Pollak, Michael; Wu, Tianying; Fuchs, Charles S; Giovannucci, Edward

    2012-06-01

    Higher serum C-peptide concentrations have shown to be associated with an increased risk of colorectal cancer (CRC). Therefore, we used diet information to identify food groups that correlated with fasting serum concentrations of C-peptide and assess the association of this dietary pattern and CRC risk. Major food contributors to fasting C-peptide concentrations were identified with stepwise linear regression in a subsample (n = 833) of women from a large cohort. We then summed the consumption frequency of the major food contributors to form a C-peptide dietary pattern for the entire cohort (n = 66,714). Risk for CRC was computed using Cox proportional hazard model with the C-peptide dietary pattern score as the predictor. In up to 20 years of follow-up, we ascertained 985 cases of CRC and 758 colon cancer. After adjusting for confounders, the C-peptide dietary pattern, characterized by higher meat, fish, and sweetened beverage intake, but lower coffee, high fat dairy, and whole grains intake, showed direct association with CRC risk (RR comparing extreme quintiles = 1.29, 95 % CI = 1.05-1.58, p trend = 0.048). The same comparison was slightly stronger for colon cancer (RR = 1.35, 95 % CI = 1.07-1.70, p trend = 0.009). In stratified analysis, there was no association between the C-peptide dietary pattern and colon cancer among lean and active women. However, for overweight or sedentary women, RR for the same comparison was 1.58 (95 % CI = 1.20-2.07, p trend = 0.002) (p for interaction = 0.007). We derived a dietary pattern that correlated with C-peptide concentrations. This pattern was associated with an increase in colon cancer, especially among women who were overweight or sedentary.

  9. Diagnostic value of C-peptide determination. [Radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Kober, G; Rainer, O H [Landeskrankenhaus Klagenfurt (Austria). Nuklearmedizinische Abt.

    1983-01-01

    C-peptide and insulin serum determinations were performed in 94 glucagon-stimulated diabetics and in 15 healthy persons. A minimal increase of 1.5 ng C-peptide/ml serum after glucagon injection (1 mg i.v.) was found to be a useful parameter for the differentiation of insulin dependent and non-insulin dependent diabetics. The maximal response to glucagon occurred during the first 10-minutes after the injection (blood was drawn at 2-minutes intervals). Serum insulin levels and basal C-peptide concentrations were of no value in predicting insulin-dependency. Basal C-peptide levels were significantly different from control in juvenile insulin dependent diabetics (decrease) only.

  10. Pregnancy-induced rise in serum C-peptide concentrations in women with type 1 diabetes

    DEFF Research Database (Denmark)

    Nielsen, Lene Ringholm; Rehfeld, Jens F; Pedersen-Bjergaard, Ulrik

    2009-01-01

    OBJECTIVE: The purpose of this study was to investigate whether pregnancy induces increased insulin production as a marker of improved beta-cell function in women with long-term type 1 diabetes. RESEARCH DESIGN AND METHODS: This was a prospective study of 90 consecutive pregnant women with type 1.......85). Multivariate regression analysis revealed a positive association between the absolute increase in C-peptide concentrations during pregnancy and decreased A1C from 8 to 33 weeks (P = 0.003). CONCLUSIONS: A pregnancy-induced increase in C-peptide concentrations in women with long-term type 1 diabetes...... in 35 women. RESULTS: C-peptide concentrations gradually increased throughout pregnancy regardless of serum glucose concentrations in the 90 women with a median duration of diabetes of 17 years (range 1-36 years). Among 35 women with paired recordings of stimulated C-peptide, C-peptide production...

  11. Some Biological Activities of Malaysian Leech Saliva Extract

    Directory of Open Access Journals (Sweden)

    Abdualrahman M. Abdualkader

    2011-12-01

    Full Text Available Normal 0 21 false false false MS X-NONE AR-SA Leeches were fed on the phagostimulatory solution through parafilm membrane. The satiated leeches were forced to regurgitate the solution by soaking them in an ice-container. The anticoagulant activity was ascertained using thrombin time assay (TT. The result revealed that the saliva concentration which increases TT by 100% (IC100 is 43.205µg/ml plasma. The antimicrobial activity of the saliva was tested against several bacterial spp. (E.coli, P.aeruginosa, B.cereus, Sal.typhi and S.aureus  and fungi spp. (C.albicans and C.neoformans. It was found that saliva has an inhibition activity against Sal.typhi (minimal inhibitory concentration MIC 78.253µg/ml, S.aureus (MIC 78.253µg/ml and E.coli (MIC 121.256µg/ml.ABSTRAK: Pacat-pacat diberi makan larutan phagostimulatory menerusi membran parafilem. Pacat-pacat yang kekenyangan itu dipaksa memuntahkan larutan tersebut dengan direndam di dalam bekas berisi ais. Aktiviti antigumpal ditentukan menggunakan cerakin masa trombin (TT. Keputusan menunjukkan kepekatan air liur pacat menyebabkan pertambahan TT sebanyak 100% (IC100 iaitu 43.205µg/ml plasma. Aktiviti antimikrob air liur telah diuji dengan pelbagai jenis bakteria (E.coli, P.aeruginosa, B.cereus, Sal.typhi dan S.aureus dan pelbagai jenis kulat (C.albicans and C.neoformans. Didapati air liur menghasilkan aktiviti perencatan terhadap Sal.typhi (kepekatan perencat minima (Minimal inhibitory concentration - MIC 78.253µg/ml, S.aureus (MIC 78.253µg/ml dan E.coli (MIC 121.256µg/ml.

  12. Unstimulated Saliva-Related Caries Risk Factors in Individuals with Cystic Fibrosis: A Cross-Sectional Analysis of Unstimulated Salivary Flow, pH, and Buffering Capacity.

    Science.gov (United States)

    Alkhateeb, Alaa A; Mancl, Lloyd A; Presland, Richard B; Rothen, Marilynn L; Chi, Donald L

    2017-01-01

    Salivary flow rate, pH, and buffering capacity are associated with dental caries, but studies from the cystic fibrosis (CF) literature are inconclusive regarding these salivary factors and caries. The aim of this study was to evaluate these factors and their associations with dental caries in individuals with CF. Unstimulated whole saliva was collected from individuals aged 6-20 years at Seattle Children's Hospital CF Clinic, USA (n = 83). Salivary flow rate was measured in milliliters per minute. Salivary pH was assessed using a laboratory pH meter. Buffering capacity was assessed by titration with HCl. The outcome measure was caries prevalence, defined as the number of decayed, missing, or filled primary and permanent tooth surfaces. Spearman's rank correlation coefficient and the t test were used to test for bivariate associations. Multiple variable linear regression models were used to (1) run confounder-adjusted analyses and (2) assess for potential interactions. There was no significant association between salivary flow rate or buffering capacity and caries prevalence. There was a significant negative association between salivary pH and caries prevalence, but this association was no longer significant after adjusting for age. There was no significant interaction between salivary flow rate and buffering capacity or between antibiotic use and the 3 salivary factors. Our results indicate that unstimulated salivary factors are not associated with dental caries prevalence in individuals with CF. Future studies should investigate other potential saliva-related caries risk factors in individuals with CF such as cariogenic bacteria levels, salivary host defense peptide levels, and medication use. © 2016 S. Karger AG, Basel.

  13. Shared and Unique Proteins in Human, Mouse and Rat Saliva Proteomes: Footprints of Functional Adaptation

    Directory of Open Access Journals (Sweden)

    Robert C. Karn

    2013-12-01

    Full Text Available The overall goal of our study was to compare the proteins found in the saliva proteomes of three mammals: human, mouse and rat. Our first objective was to compare two human proteomes with very different analysis depths. The 89 shared proteins in this comparison apparently represent a core of highly-expressed human salivary proteins. Of the proteins unique to each proteome, one-half to 2/3 lack signal peptides and probably are contaminants instead of less highly-represented salivary proteins. We recently published the first rodent saliva proteomes with saliva collected from the genome mouse (C57BL/6 and the genome rat (BN/SsNHsd/Mcwi. Our second objective was to compare the proteins in the human proteome with those we identified in the genome mouse and rat to determine those common to all three mammals, as well as the specialized rodent subset. We also identified proteins unique to each of the three mammals, because differences in the secreted protein constitutions can provide clues to differences in the evolutionary adaptation of the secretions in the three different mammals.

  14. Characterization of the cell penetrating properties of a human salivary proline-rich peptide.

    Science.gov (United States)

    Radicioni, Giorgia; Stringaro, Annarita; Molinari, Agnese; Nocca, Giuseppina; Longhi, Renato; Pirolli, Davide; Scarano, Emanuele; Iavarone, Federica; Manconi, Barbara; Cabras, Tiziana; Messana, Irene; Castagnola, Massimo; Vitali, Alberto

    2015-11-01

    Saliva contains hundreds of small proline-rich peptides most of which derive from the post-translational and post-secretory processing of the acidic and basic salivary proline-rich proteins. Among these peptides we found that a 20 residue proline-rich peptide (p1932), commonly present in human saliva and patented for its antiviral activity, was internalized within cells of the oral mucosa. The cell-penetrating properties of p1932 have been studied in a primary gingival fibroblast cell line and in a squamous cancer cell line, and compared to its retro-inverso form. We observed by mass-spectrometry, flow cytometry and confocal microscopy that both peptides were internalized in the two cell lines on a time scale of minutes, being the natural form more efficient than the retro-inverso one. The cytosolic localization was dependent on the cell type: both peptide forms were able to localize within nuclei of tumoral cells, but not in the nuclei of gingival fibroblasts. The uptake was shown to be dependent on the culture conditions used: peptide internalization was indeed effective in a complete medium than in a serum-free one allowing the hypothesis that the internalization could be dependent on the cell cycle. Both peptides were internalized likely by a lipid raft-mediated endocytosis mechanism as suggested by the reduced uptake in the presence of methyl-ß-cyclodextrin. These results suggest that the natural peptide may play a role within the cells of the oral mucosa after its secretion and subsequent internalization. Furthermore, lack of cytotoxicity of both peptide forms highlights their possible application as novel drug delivery agents.

  15. Effects of milk curd on saliva secretion in healthy volunteer compared to baseline, 2% pilocarpine and equivalent pH adjusted acetic acid solutions.

    Science.gov (United States)

    Babaee, Neda; Gholizadehpasha, Atefeh; Zahedpasha, Samir; Moghadamnia, Yasaman; Zamaninejad, Shiva; Moghadamnia, Ali Akbar

    2011-01-01

    Dry mouth is a common clinical problem, and different products have been proposed to improve it. In this investigation, the effects of "milk curd" on the amount of saliva secretion were studied. A total of 32 patients (aged 20-30) were selected from healthy volunteers. Milk curd concentrations of 0.5, 1, 2 and 4%, and 2% pilocarpine were prepared as drops. The impact of the drugs on the saliva weight was assessed after 1-5 min. To determine the effects of the pH of the milk curd on the amount of saliva secretion, different concentrations of acetic acid were used. At the end of the first minute, the differences between the data for all groups were statistically significant, and the difference between the 2% and 4% milk curd groups was higher than the others (P saliva secreted at the end of the second minute between the baseline and 4% milk curd groups and between the 0.5% and 4% MC groups were significant (P = 0.006 and P = 0.025, respectively). In total, there was no significant difference between the effect of various pH treatments and the amount of baseline saliva secretion. Milk curd has a significant local impact, and the saliva increase depends on the dose. It seems that this effect is not only related to its acidic taste. As a result, factors other than pH are involved in the effect.

  16. Efek Pengunyahan Permen Karet Gula dan Xylitol terhadap Status Saliva

    Directory of Open Access Journals (Sweden)

    Lisna Kurnia Rezky

    2016-11-01

    Full Text Available Latar belakang. Rongga mulut sebagai pintu masuk makanan ke dalam tubuh selalu dibasahi oleh saliva setiap harinya. Saat ini banyak produk permen karet yang beredar di masyarakat yang mengandung gula dan xylitol. Banyak orang yang gemar mengunyah permen karet dengan kurang memperhatikan komposisinya baik yang mengandung gula ataupun xylitol sehingga kurang mengetahui efek masing-masing jenis permen karet tersebut terhadap kesehatan rongga mulut. Tujuan. Penelitian ini bertujuan untuk mengetahui efek pengunyahan permen karet gula dengan permen karet xylitol terhadap status saliva yang terdiri dari volume, pH, dan viskositas saliva. Metode penelitian. Subjek penelitian berjumlah 30 orang dibagi menjadi 3 kelompok masing-masing 10 orang, terdiri dari kelompok mengunyah permen karet gula, xylitol, dan kontrol dengan mengunyah apel. Pengambilan saliva dilakukan pagi hari dan siang hari. Subjek mengunyah 2 butir permen karet dan tidak diperbolehkan untuk makan dan minum 1 jam sebelum mengunyah. Subjek diinstruksikan meludah ke dalam pot saliva selama 10 menit dalam interval setiap 1 menit. Pengukuran volume saliva menggunakan pipet volume, pH saliva dengan menggunakan pH meter, dan viskositas saliva dengan menggunakan viskometer Ostwald hari ke-1 dan ke-4. Analisis data dengan uji statistik Mann-Whitney. Hasil. penelitian menunjukkan adanya peningkatan bermakna volume dan viskositas saliva pada pengunyahan permen karet xylitol dan gula. Derajat keasaman (pH saliva menurun setelah mengunyah permen karet gula sedangkan pada perm en karet xylitol relatif stabil. Disimpulkan bahwa permen karet xylitollebih baik untuk kestabilan status saliva dibandingkan permen karet gula.

  17. Influence of artificial saliva in biofilm formation of Candida albicans in vitro

    Directory of Open Access Journals (Sweden)

    Michelle Peneluppi Silva

    2012-02-01

    Full Text Available Due to the increase in life expectancy, new treatments have emerged which, although palliative, provide individuals with a better quality of life. Artificial saliva is a solution that contains substances that moisten a dry mouth, thus mimicking the role of saliva in lubricating the oral cavity and controlling the existing normal oral microbiota. This study aimed to assess the influence of commercially available artificial saliva on biofilm formation by Candida albicans. Artificial saliva I consists of carboxymethylcellulose, while artificial saliva II is composed of glucose oxidase, lactoferrin, lysozyme and lactoperoxidase. A control group used sterile distilled water. Microorganisms from the oral cavity were transferred to Sabouraud Dextrose Agar and incubated at 37°C for 24 hours. Colonies of Candida albicans were suspended in a sterile solution of NaCl 0.9%, and standardisation of the suspension to 106 cells/mL was achieved. The acrylic discs, immersed in artificial saliva and sterile distilled water, were placed in a 24-well plate containing 2 mL of Sabouraud Dextrose Broth plus 5% sucrose and 0.1 mL aliquot of the Candida albicans suspension. The plates were incubated at 37°C for 5 days, the discs were washed in 2 mL of 0.9% NaCl and placed into a tube containing 10 mL of 0.9% NaCl. After decimal dilutions, aliquots of 0.1 mL were seeded on Sabouraud Dextrose Agar and incubated at 37°C for 48 hours. Counts were reported as CFU/mL (Log10. A statistically significant reduction of 29.89% (1.45 CFU/mL of Candida albicans was observed in saliva I when compared to saliva II (p = 0.002, considering p≤0.05.

  18. Uncovering the Peptide-Binding Specificities of HLA-C: A General Strategy To Determine the Specificity of Any MHC Class I Molecule

    DEFF Research Database (Denmark)

    Rasmussen, Michael; Harndahl, Mikkel; Stryhn, Anette

    2014-01-01

    MHC class I molecules (HLA-I in humans) present peptides derived from endogenous proteins to CTLs. Whereas the peptide-binding specificities of HLA-A and -B molecules have been studied extensively, little is known about HLA-C specificities. Combining a positional scanning combinatorial peptide...... library approach with a peptide-HLA-I dissociation assay, in this study we present a general strategy to determine the peptide-binding specificity of any MHC class I molecule. We applied this novel strategy to 17 of the most common HLA-C molecules, and for 16 of these we successfully generated matrices...... representing their peptide-binding motifs. The motifs prominently shared a conserved C-terminal primary anchor with hydrophobic amino acid residues, as well as one or more diverse primary and auxiliary anchors at P1, P2, P3, and/or P7. Matrices were used to generate a large panel of HLA-C-specific peptide...

  19. A device for the collection of submandibular saliva.

    Science.gov (United States)

    Hanning, Sara; Motoi, Lidia; Medlicott, Natalie; Swindells, Stephen

    2012-03-01

    The objective of this study was to describe the construction of a non-invasive device for the collection of submandibular saliva. Preliminary tests were carried out on saliva collected from a single donor in order to determine whether the rheological properties of submandibular saliva collected using the device were comparable to whole saliva collected using the expectoration (or 'spit') method. The device collected a lower quantity of saliva than that collected using the expectoration method. Stimulated saliva collected using the device had a pH close to that of unstimulated saliva because the sealed collection unit in the device minimised contamination. Saliva exhibited shear-thinning behaviour regardless of the method of collection, although that collected using the device was more viscous. The viscoelasticity of saliva collected using the two methods was different, probably as a result of differences in composition. This difference was greater with stimulated saliva. Despite the discrepancies between whole saliva and submandibular saliva, the device provides a non-invasive method for the collection of high-quality saliva over extended periods.

  20. Study on the C-peptide radioimmunoassay with synthetized connecting peptide

    Energy Technology Data Exchange (ETDEWEB)

    Nakagawa, S; Sasaki, T; Nakayama, H; Watanabe, T; Aoki, S [Hokkaido Univ., Sapporo (Japan). School of Medicine

    1976-01-01

    A method of C-peptide radioimmunoassay with the synthetized connecting peptide by Yanaihara was tested for the determination of serum C-peptide immunoreactivity (CPR) in normal people and in diabetics with or without insulin treatment. The CPR value obtained by this method was not interfered with by the presence of serum proteins or by the insulin of people with or without insulin treatment judged by the dilution test and the recovery test. The normal fasting CPR was 2.80 +- 0.78 ng/ml with the synthetized C-peptide as a standard. The CPR value increased and reached a maximum 90 minutes after the ingestion of 50 g of glucose. The increase after the glucose loading reduced corresponding to the severity of diabetes, and some juvenile-onset diabetes showed no response. Adult-type diabetics under insulin treatment, however, showed weak but significant CPR response. The increment of CPR and immunoreactive insulin after glucose loading in normal people and non-treated diabetics was well correlated (..gamma..=0.8262). Judged from the above mentioned results, CPR determination in insulin-treated diabetics was thought to be a useful method for the assessment of the insulin-secreting ability of beta-cells of the pancreas.

  1. Effects of milk curd on saliva secretion in healthy volunteer compared to baseline, 2% pilocarpine and equivalent pH adjusted acetic acid solutions

    Directory of Open Access Journals (Sweden)

    Neda Babaee

    2011-01-01

    Full Text Available Background: Dry mouth is a common clinical problem, and different products have been proposed to improve it. In this investigation, the effects of "milk curd" on the amount of saliva secretion were studied. Materials and Methods: A total of 32 patients (aged 20-30 were selected from healthy volunteers. Milk curd concentrations of 0.5, 1, 2 and 4%, and 2% pilocarpine were prepared as drops. The impact of the drugs on the saliva weight was assessed after 1-5 min. To determine the effects of the pH of the milk curd on the amount of saliva secretion, different concentrations of acetic acid were used. Results: At the end of the first minute, the differences between the data for all groups were statistically significant, and the difference between the 2% and 4% milk curd groups was higher than the others (P < 0.0001. The differences in the amount of the saliva secreted at the end of the second minute between the baseline and 4% milk curd groups and between the 0.5% and 4% MC groups were significant (P = 0.006 and P = 0.025, respectively. In total, there was no significant difference between the effect of various pH treatments and the amount of baseline saliva secretion. Conclusion: Milk curd has a significant local impact, and the saliva increase depends on the dose. It seems that this effect is not only related to its acidic taste. As a result, factors other than pH are involved in the effect.

  2. Dynamic changes in saliva after acute mental stress

    Science.gov (United States)

    Naumova, Ella A.; Sandulescu, Tudor; Bochnig, Clemens; Khatib, Philipp Al; Lee, Wing-Kee; Zimmer, Stefan; Arnold, Wolfgang H.

    2014-01-01

    Stress-related variations of fluoride concentration in supernatant saliva and salivary sediment, salivary cortisol, total protein and pH after acute mental stress were assessed. The hypothesis was that stress reactions have no influence on these parameters. Thirty-four male students were distributed into two groups: first received the stress exposure followed by the same protocol two weeks later but without stress exposure, second underwent the protocol without stress exposure followed by the stress exposure two weeks later. The stressor was a public speech followed by tooth brushing. Saliva was collected before, immediately after stress induction and immediately, at 10, 30 and 120 min. after tooth brushing. Cortisol concentrations, total protein, intraoral pH, and fluoride content in saliva were measured. The data were analyzed statistically. Salivary sediment was ca 4.33% by weight of whole unstimulated saliva. Fluoride bioavailability was higher in salivary sediment than in supernatant saliva. The weight and fluoride concentration was not altered during 2 hours after stress exposure. After a public speech, the salivary cortisol concentration significantly increased after 20 minutes compared to the baseline. The salivary protein concentration and pH also increased. Public speaking influences protein concentration and salivary pH but does not alter the fluoride concentration of saliva. PMID:24811301

  3. Epimerization-free C-terminal peptide activation, elongation and cyclization

    NARCIS (Netherlands)

    Popović, S.

    2015-01-01

    C-terminal peptide activation and cyclization reactions are generally accompanied with epimerization (partial loss of C‐terminal stereointegrity). Therefore, the focus of this thesis was to develop epimerization-free methods for C-terminal peptide activation to enable C-terminal peptide elongation

  4. [Saliva: more than just a factor in oral health].

    Science.gov (United States)

    van Nieuw Ameronpn, A; Veerman, E C I; Brand, H S; Vissink, A

    2006-05-01

    Several salivary proteins are involved in the antimicrobial defence within the oral cavity, and are able to eliminate or inhibit a great number of microorganisms that enter the body via the oral cavity. Thus changes in the composition of saliva may not only affect people's oral health, but also have consequences for their general health. Based on salivary proteins, antimicrobial peptides have been synthesized. Potentially, these could be used as a new class of antibiotics. The antimicrobial peptides are hardly cytotoxic to human cells and do not evoke any or only very little resistence in microorganisms.

  5. pH dependence of MHC class I-restricted peptide presentation

    DEFF Research Database (Denmark)

    Stryhn, A; Pedersen, L O; Romme, T

    1996-01-01

    The function of MHC class I molecules is to bind and present antigenic peptides to cytotoxic T cells. Here, we report that class I-restricted peptide presentation is strongly pH dependent. The presentation of some peptides was enhanced at acidic pH, whereas the presentation of others was inhibited....... Biochemical peptide-MHC class I binding assays demonstrated that peptide-MHC class I complexes are more stable at neutral pH than at acidic pH. We suggest that acid-dependent peptide dissociation can generate empty class I molecules and that the resulting binding potential can be exploited by a subset...

  6. Preventive effects of the novel antimicrobial peptide Nal-P-113 in a rat Periodontitis model by limiting the growth of Porphyromonas gingivalis and modulating IL-1β and TNF-α production.

    Science.gov (United States)

    Wang, Hong-Yan; Lin, Li; Fu, Wei; Yu, Hui-Yuan; Yu, Ning; Tan, Li-Si; Cheng, Jya-Wei; Pan, Ya-Ping

    2017-08-29

    P-113 (AKRHHGYKRKFH-NH2) is a 12-amino-acid histidine-rich peptide derived from histatin 5 that is highly degradable in high salt concentrations and biological fluids such as serum, plasma and saliva. Nal-P-113, a novel antimicrobial peptide whose histidine residues are replaced by the bulky amino acids β-naphthylalanine, causes the antimicrobial peptide to retain its bactericidal activity even in physiological environments. This study evaluated the effect of the novel antimicrobial peptide Nal-P-113 in a rat periodontitis model and the mechanisms of action of Nal-P-113 for suppressing periodontitis. Periodontitis was induced in mandibular first molars in rats receiving a ligature and infected with Porphyromonas gingivalis. Animals were randomly divided into six groups: a, P. gingivalis W83 alone; b, P. gingivalis W83 with 6.25 μg/mL of Nal-P-113; c, P. gingivalis W83 with 25 μg/mL of Nal-P-113; d, P. gingivalis W83 with 100 μg/mL of Nal-P-113; e, P. gingivalis W83 with 400 μg/mL of Nal-P-113; and f, control without P. gingivalis W83 or Nal-P-113. Morphometric analysis was used to evaluate alveolar bone loss. Microbiological assessment of the presence of Porphyromonas gingivalis and total bacteria was performed using absolute quantitative real-time PCR and scanning electron microscopy. Gingival tissue was collected for western blot and immunohistochemical assays of IL-1β and TNF-α levels. Alveolar bone loss was inhibited by 100 μg/mL or 400 μg/mL of Nal-P-113 compared to the control group (P periodontal tissue (P periodontitis in rats by limiting the amount of bacteria and modulating IL-1β and TNF-α production. The use of Nal-P-113 in vivo might serve as a beneficial preventive or therapeutic approach for periodontitis.

  7. Insulin and C peptide response, and antibody levels in hepatitis C related chronic liver disease

    International Nuclear Information System (INIS)

    Abbas, Z.; Tariq, N.; Iqbal, M.; Shah, M.A.

    2002-01-01

    Objective: Patients with cirrhosis due to hepatitis C (HC) have an increased prevalence of diabetes mellitus. The pathogenic mechanism by which HC predisposes to DM is not clear. The objective of this study was to determine the insulin and C-peptide response to 75 gram oral glucose load and measure anti phospholipid antibody levels in patients with chronic liver disease due to HC. Design: a prospective study. Place and duration of study: This study was conducted at the department of medicine, Jinnah postgraduate medical centre over period of three months. Subjects and methods: An analytical case control study was carried out on 37 patients (m-18,f=19); none of these patients had received interferon. They were divided into four groups: (a) HC cirrhosis with DM (n=9 ), (b) HC cirrhosis without DM (n=11), (c) hepatitis B (HB) cirrhosis without DM (n=7), (d) chronic hepatitis C without DM (n=10). Group C and D were taken as controls. Fasting blood samples were taken and repeated after 2 hours of 75 gram oral glucose load (2 h PG). Result: mean ages of group A,B,C and D were (yr +- SD) 51.3 +- 7.6,48.9 +- 2.4, 33.7 +-10.8 and 31.7 +- 8.8 respectively. There was no statistically significant difference in the age, Pugh score and body mass index of HC cirrhotic patients with and without DM. Patients of group A had higher fasting and 2 h PG glucose levels (P=0.003 and 0.000) and higher fasting insulin level (p=0.045). However, increments in insulin and c peptide levels 2 h PG were much less (p=0.048 and 0.003). HB cirrhotics without diabetes (group C behaved just like HC cirrhotic without diabetes (group B). Patients of group D had normal glucose tolerance and insulin and C peptide levels. All four groups had normal anti phospholipid antibody levels. Conclusion: Patients with cirrhosis due to HC nd HB show evidence of glucose intolerance in spite of hyperinsulinaemia probably due to insulin resistance. HC cirrhotics with diabetes have fasting hyperglycemia in spite of

  8. Fundamental studies on the development of C-peptide radioimmunoassay kit

    International Nuclear Information System (INIS)

    Nakazawa, Nobuhiko; Maki, Kentaro; Ogawa, Hiroshi; Ikeda, Osamu

    1976-01-01

    We have studied the development of the C-peptide radioimmunoassay kit which is usable in the pancreatic function test with satisfactory results. The C-peptide antiserum was prepared by immunizing rabbits with synthetic human connecting peptide. The antiserum revealed no cross reaction with any C-peptides other than human C-peptide, porcine insulin and gastrointestinal hormone, and showed high specificity to human C-peptide. We adopted the double antibody method in B,F separation, and chose 4 0 C, 48 hrs. for 1st. incubation and 4 0 C, 24 hrs. for 2nd. incubation. On this kit, the assay range was 0.5 ng/ml-30 ng/ml, the recovery rate was 98.4%-107.8% in the recovery test, the coefficient of variance was 6.2% in the intra assay and was 7.6% in the inter assay. We think this kit is sufficiently usable to assay C-peptide in blood. (auth.)

  9. Fundamental studies on the development of C-peptide radioimmunoassay kit

    Energy Technology Data Exchange (ETDEWEB)

    Nakazawa, N; Maki, K; Ogawa, H; Ikeda, O [Daiichi Radioisotope Labs. Ltd., Tokyo (Japan)

    1976-05-01

    We have studied the development of the C-peptide radioimmunoassay kit which is usable in the pancreatic function test with satisfactory results. The C-peptide antiserum was prepared by immunizing rabbits with synthetic human connecting peptide. The antiserum revealed no cross reaction with any C-peptides other than human C-peptide, porcine insulin and gastrointestinal hormone, and showed high specificity to human C-peptide. We adopted the double antibody method in B,F separation, and chose 4/sup 0/C, 48 hrs. for 1st. incubation and 4/sup 0/C, 24 hrs. for 2nd. incubation. On this kit, the assay range was 0.5 ng/ml-30 ng/ml, the recovery rate was 98.4%-107.8% in the recovery test, the coefficient of variance was 6.2% in the intra assay and was 7.6% in the inter assay. We think this kit is sufficiently usable to assay C-peptide in blood.

  10. Immunological half-life of porcine proinsulin C-peptide

    Energy Technology Data Exchange (ETDEWEB)

    Oyama, H; Horino, M; Matsumura, S [Kawasaki Medical Coll., Kurashiki (Japan). Div. of Endocrinology; Kobayshi, K; Suetsugu, N [Yamaguchi Univ., Ube (Japan). School of Medicine

    1975-11-01

    Immunological half-lifes of injected porcine C-peptide and insulin with RIA were studied and calculated as 9.8 and 8.0 minutes. Higher circulating levels of C-peptide as compared to insulin in normal young swines lead to speculation about a longer half-life of C-peptide. This hypothesis was verified in this study. Immunological half-lifes of porcine proinsulin and insulin in the pig were 20 and 6 minutes, respectively.

  11. Preserved C-peptide levels in overweight or obese compared with underweight children upon diagnosis of type 1 diabetes mellitus

    Directory of Open Access Journals (Sweden)

    Hyeoh Won Yu

    2015-06-01

    Full Text Available PurposeWe hypothesized that overweight or obese children might develop type 1 diabetes mellitus (T1DM early despite residual beta-cell function. Factors independently associated with preservation of C-peptide level were analyzed.MethodsWe retrospectively reviewed the medical data of 135 children aged 2.1-16.5 years with autoimmune T1DM. Body mass index (BMI, pubertal stage, and glycosylated hemoglobin (HbA1c and C-peptide levels were evaluated. Patients were assigned to underweight (22.2%, normal weight (63.7%, and overweight or obese (14.1% groups according to their BMI.ResultsPreservation of serum C-peptide levels (≥0.6 ng/mL was found in 43.0% of subjects. With increasing BMI, the proportions of children with preserved C-peptide levels increased from 33.3% to 41.9% to 63.2%, with marginal significance (P=0.051. Interaction analysis indicated no effect of BMI score on age at onset associated with serum C-peptide levels. The lower the C-peptide level, the younger the age of onset (P<0.001, after adjustment for BMI z-score and HbA1c level. However, no significant relationship between BMI z-score or category and onset age was evident. Upon multivariate-adjusted modeling, the odds that the C-peptide level was preserved increased by 1.2 fold (P=0.001 per year of life, by 3.1 folds (P=0.015 in children presenting without (compared to with ketoacidosis, and by 5.0 folds (P=0.042 in overweight or obese (compared to underweight children.ConclusionOverweight or obese children had slightly more residual beta-cell function than did underweight children. However, we found no evidence that obesity temporally accelerates T1DM presentation.

  12. Treatment of Oral Multispecies Biofilms by an Anti-Biofilm Peptide.

    Science.gov (United States)

    Wang, Zhejun; de la Fuente-Núñez, Cesar; Shen, Ya; Haapasalo, Markus; Hancock, Robert E W

    2015-01-01

    Human oral biofilms are multispecies microbial communities that exhibit high resistance to antimicrobial agents. Dental plaque gives rise to highly prevalent and costly biofilm-related oral infections, which lead to caries or other types of oral infections. We investigated the ability of the recently identified anti-biofilm peptide 1018 to induce killing of bacterial cells present within oral multispecies biofilms. At 10 μg/ml (6.5 μM), peptide 1018 was able to significantly (pbiofilm formation over 3 days. The activity of the peptide on preformed biofilms was found to be concentration-dependent since more than 60% of the total plaque biofilm cell population was killed by 10 μg/ml of peptide 1018 in 3 days, while at 5 μg/ml 50% of cells were dead and at 1 μg/ml the peptide triggered cell death in around 30% of the total bacterial population, as revealed by confocal microscopy. The presence of saliva did not affect peptide activity, since no statistically significant difference was found in the ability of peptide 1018 to kill oral biofilms using either saliva coated and non-saliva coated hydroxyapatite surfaces. Scanning electron microscopy experiments indicated that peptide 1018 induced cell lysis in plaque biofilms. Furthermore, combined treatment using peptide 1018 and chlorhexidine (CHX) increased the anti-biofilm activity of each compound compared to when these were used alone, resulting in >50% of the biofilm being killed and >35% being dispersed in only 3 minutes. Peptide 1018 may potentially be used by itself or in combination with CHX as a non-toxic and effective anti-biofilm agent for plaque disinfection in clinical dentistry.

  13. Changes of serum leptin and c-peptide level in children with type 1 diabetic mellitus

    International Nuclear Information System (INIS)

    Du Tongxin; Wang Zizheng; Sun Junjiang; Wang Shukui; Qi Shaokang

    2001-01-01

    To deplore the relationship between leptin and c-peptide in children with type 1 diabetic mellitus (DM). The levels of serum leptin and c-peptide (C-P) in 65 type 1 DM children (including 31 before and after insulin treatment) and 30 normal controls were measured by radioimmunoassay (RIA). The results found that there was significant differences (P < 0.01) in leptin and C-P between DM children and normal controls, also in 31 DM children before and after treatment. It showed a positive correlation between leptin and C-P. The changes of the leptin/C-P ratio in DM children compared with normal controls and that before and after treatment were also significantly different. It suggested that leptin may have close relationship in the development, progress and the occurrence of complications in children with DM and also provide a new clue for their diagnosis treatment and complication occurrence

  14. Zein nanoparticle as a novel BMP6 derived peptide carrier for enhanced osteogenic differentiation of C2C12 cells.

    Science.gov (United States)

    Hadavi, Mahvash; Hasannia, Sadegh; Faghihi, Shahab; Mashayekhi, Farhad; Homazadeh, Homayoun; Mostofi, Seyed Behrooz

    2018-01-26

    Zein nanoparticles as a carrier system for BMP6-derived peptide were prepared by liquid-liquid phase separation procedure and characterized with SEM, DLS, FTIR and thermogravimetric methods. After peptide encapsulation, nanoparticle size increased from 236.3 ± 92.2 nm to 379.4 ± 116.8 nm. The encapsulation efficiency of peptide was 72.6% and the release of peptide from Zein nanoparticles was partly sustained in trypsin containing phosphate buffered saline (pH 7.4) for up to 14 days. Peptide-loaded nanoparticles showed similar cell viability compared with blank ones. ALP activity of C2C12 cells treated with peptide-loaded nanoparticles (500 µg/mL) was evaluated 7, 14, 21 and 28 days after culture. In peptide-loaded nanoparticles, ALP activity was significantly higher (p < .05) compared with other groups at day 14. Alizarin Red S staining showed, C2C12 cells behind peptide-loaded nanoparticles had significantly (p < .05) higher calcium deposition at day 21. The results of RT-qPCR show that the BMP-6 peptide activated expression of RUNX2 as a transcription factor. In turn, RUNX2 regulates SPP1 and BGLAP gene expression, as osteogenic marker genes. The results confirm that the peptide-loaded Zein nanoparticles, as osteoinductive material, may be used to repair small area of bone defects, with low load bearing.

  15. Factors That Influence the Extensional Rheological Property of Saliva.

    Directory of Open Access Journals (Sweden)

    Amrita Vijay

    Full Text Available The spinnbarkeit of saliva reflects the ability of saliva to adhere to surfaces within the mouth, thereby serving as a protective role and aiding in lubrication. Therefore, alterations in the extensional rheology of saliva may result in the loss in adhesiveness or the ability to bind onto surfaces. Mucin glycoproteins and their structures are known to be important factors for the extensional rheological properties of saliva. The conformation of mucin depends on factors such as pH and ionic strength. Chewing is one of the main stimuli for salivary secretion but creates significant sheer stress on the salivary film which could influence mouthfeel perceptions. The current study investigates the possible factors which affect the extensional rheological properties of saliva by comparing submandibular/sublingual saliva with different oral stimuli within the same group of subjects. Unstimulated and stimulated saliva (chew, smell and taste salivas were collected primarily from submandibular/sublingual glands. The saliva samples were measured for Spinnbarkeit followed by the measuring mucin, total protein, total calcium and bicarbonate concentrations. The results indicated correlations between rheological properties and mucin/ion concentrations. However, chewing stimulated submandibular/sublingual saliva is shown to have significantly lower Spinnbarkeit, but factors such as mucin, protein and calcium concentrations did not account for this variation. Analysis of the concentration of bicarbonate and pH appears to suggest that it has a prominent effect on extensional rheology of saliva.

  16. Preliminary findings on the correlation of saliva pH, buffering capacity, flow rate and consistency in relation to waterpipe tobacco smoking

    OpenAIRE

    Khemiss, Mehdi; Ben Khelifa, Mohamed; Ben Saad, Helmi

    2017-01-01

    ABSTRACT The aim of the present comparative study was to compare some salivary characteristics between exclusive waterpipe smokers (EWPS) and non-smokers. 72 males (36 EWPS) were recruited. The volume of stimulated saliva was determined and divided by the duration of saliva collection. The pH was measured directly using a pH meter. The buffering capacity was determined using a quantitative method which involved the addition of 10 ?l HCl. Up to a total of 160 ?L was titrated up to obtain a pH ...

  17. Effect of saliva contamination on the microshear bond strength of one-step self-etching adhesive systems to dentin.

    Science.gov (United States)

    Yoo, H M; Oh, T S; Pereira, P N R

    2006-01-01

    This study evaluated the effect of saliva contamination and decontamination methods on the dentin bond strength of one-step self-etching adhesive systems. Three commercially available "all-in-one" adhesives (One Up Bond F, Xeno III and Adper Prompt) and one resin composite (Filtek Z-250) were used. Third molars stored in distilled water with 0.5% thymol at 4 degrees C were ground with #600 SiC paper under running water to produce a standardized smear layer. The specimens were randomly divided into groups according to contamination methods: no contamination, which was the control (C); contamination of the adhesive surface with fresh saliva before light curing (A) and contamination of the adhesive surface with fresh saliva after light curing (B). Each contamination group was further subdivided into three subgroups according to the decontamination method: A1-Saliva was removed by a gentle air blast and the adhesive was light-cured; A2-Saliva was rinsed for 10 seconds, gently air-dried and the was adhesive light-cured; A3-Saliva was rinsed and dried as in A2, then the adhesive was re-applied to the dentin surface and light-cured; B1-Saliva was removed with a gentle air blast; B2-Saliva was rinsed and dried; B3-Saliva was rinsed, dried and the adhesive was re-applied and light cured. Tygon tubes filled with resin composite were placed on each surface and light cured. All specimens were stored in distilled water at 37 degrees C for 24 hours. Microshear bond strength was measured using a universal testing machine (EZ test), and data were analyzed by one-way ANOVA followed by the Duncan test to make comparisons among the groups (p0.05). Bond strengths of all B groups were significantly lower compared to the controls (pcontamination after adhesive curing. There was no statistically significant difference among the control groups (p>0.05).

  18. The association between newborn regional body composition and cord blood concentrations of C-peptide and insulin-like growth factor I

    DEFF Research Database (Denmark)

    Carlsen, Emma M; Renault, Kristina M; Jensen, Rikke B

    2015-01-01

    BACKGROUND: Third trimester fetal growth is partially regulated by C-peptide and insulin-like growth factor I (IGF-I). Prenatal exposures including maternal obesity and high gestational weight gain as well as high birth weight have been linked to subsequent metabolic disease. We evaluated...... with both C-peptide (p tissue accumulation was associated with cord blood C-peptide and IGF-I. Total and abdominal fat masses were related to C-peptide but not to IGF-I. Thus, newborn adiposity is partially mediated through C-peptide and early...

  19. C-terminal peptides of tissue factor pathway inhibitor are novel host defense molecules.

    Science.gov (United States)

    Papareddy, Praveen; Kalle, Martina; Kasetty, Gopinath; Mörgelin, Matthias; Rydengård, Victoria; Albiger, Barbara; Lundqvist, Katarina; Malmsten, Martin; Schmidtchen, Artur

    2010-09-03

    Tissue factor pathway inhibitor (TFPI) inhibits tissue factor-induced coagulation, but may, via its C terminus, also modulate cell surface, heparin, and lipopolysaccharide interactions as well as participate in growth inhibition. Here we show that C-terminal TFPI peptide sequences are antimicrobial against the gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa, gram-positive Bacillus subtilis and Staphylococcus aureus, as well as the fungi Candida albicans and Candida parapsilosis. Fluorescence studies of peptide-treated bacteria, paired with analysis of peptide effects on liposomes, showed that the peptides exerted membrane-breaking effects similar to those seen for the "classic" human antimicrobial peptide LL-37. The killing of E. coli, but not P. aeruginosa, by the C-terminal peptide GGLIKTKRKRKKQRVKIAYEEIFVKNM (GGL27), was enhanced in human plasma and largely abolished in heat-inactivated plasma, a phenomenon linked to generation of antimicrobial C3a and activation of the classic pathway of complement activation. Furthermore, GGL27 displayed anti-endotoxic effects in vitro and in vivo in a mouse model of LPS shock. Importantly, TFPI was found to be expressed in the basal layers of normal epidermis, and was markedly up-regulated in acute skin wounds as well as wound edges of chronic leg ulcers. Furthermore, C-terminal fragments of TFPI were associated with bacteria present in human chronic leg ulcers. These findings suggest a new role for TFPI in cutaneous defense against infections.

  20. Measurement of insulin and C-peptide excitatory test levels in gestational diabetes mellitus

    International Nuclear Information System (INIS)

    Du Tongxin; Wang Zizheng

    2001-01-01

    To investigate the function of islet β cells in patients with gestational diabetes mellitus (GDM), serum insulin and C-peptide (C-P) excitatory test levels were measured dynamically by radioimmunoassay in 41 patients with GDM and 30 normal pregnant controls. The results showed that there were significant difference in insulin and C-peptide excitatory test levels between normal pregnancy for 32-40 weeks and patients with GDM (P < 0.001). The secretory peak of insulin occurred at 60 min in normal pregnancy, while at 120 min in patients with GDM, and the recovery postponed in patients with GDM. The peak time for C-P was just as same as that of insulin, but the peak error for C-P between normal pregnant controls and patients with GDM was more larger than that for insulin and it recovered more slowly. It suggested that majority of islet β cells in patients with GDM were good enough for response to islet resistance factors and big stress from pregnancy, and also suggested a relation between pregnancy and islet β cells function

  1. Immunogenicity Evaluation of A Synthetic Peptide EgP-29aa134-142 from Echinococcus granulosus In BALB /c Strain Mice

    Directory of Open Access Journals (Sweden)

    Sh. Jafari

    2016-02-01

    Full Text Available Background: Cystic echinococcosis or hydatid disease is a widely endemic helminthic  zoonotic disease caused by infection with metacestodes the larval stage of the tapeworm Echinococcus granulosus. P-29, a 29-kDa antigen from E. granulosus, is a protoscolex specific component. The immunologic cross reactivity between P-29 and a major diagnostic antigen of E. granulosus (Ag5 indicated that P-29 might be another useful antigen of E. granulosus to be used in diagnosis or in multi epitope vaccines to prevent secondary echinococcosis. In this study, the peptide epitope regions after the identification and synthesis were evaluated in order to determine the effect of stimulating an immune response in a mouse model. Methods: In this study, the p29 antigen epitope regions of the parasite Echinococcus granulosus detected by the IEDB Bioinformatics software and a 8 amino acids sequence were synthesized. BALB/c mice were immunized subcutaneously three times with two weeks interval. Fourteen days after last immunization spleen tissues were extracted and splenocytes were cultured in presence of antigen for 72 hours. Supernatants were collected and used for cytokine assay by Quantikine  ELISA kit. Results: Sandwich ELISA results were analyzed and showed significant difference in IFN-γ but no significant differences observed in levels of two cytokines IL-4, IL-10, between immunized and control groups. Conclusion: This study has assessed the effect of synthetic peptide EgP-29aa134-142 on the immune response and our results showed that the peptide can increase IFN-γ and therefore induce activation of innate Immunity as well as CTL and Th1 response.

  2. Frequency of enterococcus faecalis in saliva and root canals with treatment failure

    International Nuclear Information System (INIS)

    Khan, I.; Shan, T.; Manzoor, M.A.

    2015-01-01

    To compare the frequency of E. faecalis in the saliva and root canals of teeth associated with apical periodontitis due to endodontic treatment failure in the same patient. Study Design: Cross-sectional comparative study. Place and Duration of Study: Samples were collected from Operative Dentistry Department, AFID, while laboratory processing was done at AFIP, Rawalpindi. Study duration was one year. Patients and Methods: Fifty patients, both males and females with failed endodontic treatment were selected. Saliva and root canal samples were collected from each patient, inoculated on MacKonkey agar plate and incubated at 35-37 degree C for 48 hrs. E. faecalis colonies were identified by colony morphology, gramstain, catalase, bile asculin test, arabinose fermentation and growth in 6% NaCl nutrient broth. Results: The frequency of E. faecalis in saliva was 34% and in root canal it was 58%. Frequency between the presence of E. faecalis in root canals and saliva was found to be statistically different (p = 0.001). Conclusion: The presence of E. faecalis in root canal was not associated with their presence in saliva. (author)

  3. Frequency of enterococcus faecalis in saliva and root canals with treatment failure

    International Nuclear Information System (INIS)

    Shan, T.; Manzoor, M.A.; Hussain, W.

    2014-01-01

    To compare the frequency of E.faecalis in the saliva and root canals of teeth associated with apical periodontitis due to endodontic treatment failure Study. Design: Cross-sectional comparative. Place and Duration of Study: Samples were collected from Operative Dentistry department, AFID, while laboratory processing was done at AFIP, Rawalpindi. Duration of this study was one year. Patients and Method: Fifty patients, both males and females with failed endodontic treatment were selected. Saliva and root canal samples were collected from each patient, inoculated on MacKonkey agar plate and incubated at 35-370 C for 48 hours. E.faecalis colonies were identified by colony morphology, Gram stain, catalase, bile asculin test, arabinose fermentation and growth in 6% NaCl nutrient broth. Results: The frequency of E.faecalis in saliva was 34% and 58% in root canal samples. Frequency of the presence of E.faecalis in root canals and saliva was found to be statistically different (p=0.000). Conclusion: The presence of E.faecalis in root canal was not associated with their presence in saliva. (author)

  4. Radioimmunological determination of chloramphenicol in the saliva of lactating cows

    International Nuclear Information System (INIS)

    Dotter, A.; Kroker, R.; Arnold, D.; Somogyi, A.

    1987-01-01

    In an effort to search for noninvasive methods suitable to monitor compliance with the ban of chloramphenicol (CAP) in milkproducing animals, the pharmacokinetic behavior of this drug in bovine saliva was investigated. As revealed by studies using a radioimmunological assay, CAP appears following its intracisternal (i.c.) or subcutaneous (s.c.) administration in the saliva of lactating cows. The level of sensitivity of the method (1.5 ng CAP per g saliva) was reached 14 and 18 days after i.c. and s.c. administration, respectively. At present, the question must remain open as to whether the concentration of CAP in the saliva can serve as a reliable indicator for the enforcement of the highest permissible level set at 1 ng CAP per g of milk by German regulations. (orig.) [de

  5. Radioimmunological determination of chloramphenicol in the saliva of lactating cows

    Energy Technology Data Exchange (ETDEWEB)

    Dotter, A.; Kroker, R.; Arnold, D.; Somogyi, A.

    1987-02-01

    In an effort to search for noninvasive methods suitable to monitor compliance with the ban of chloramphenicol (CAP) in milk-producing animals, the pharmacokinetic behavior of this drug in bovine saliva was investigated. As revealed by studies using a radioimmunological assay, CAP appears following its intracisternal (i.c.) or subcutaneous (s.c.) administration in the saliva of lactating cows. The level of sensitivity of the method (1.5 ng CAP per g saliva) was reached 14 and 18 days after i.c. and s.c. administration, respectively. At present, the question must remain open as to whether the concentration of CAP in the saliva can serve as a reliable indicator for the enforcement of the highest permissible level set at 1 ng CAP per g of milk by German regulations.

  6. C-type natriuretic peptide and its precursor

    DEFF Research Database (Denmark)

    Lippert, Solvej; Iversen, Peter; Brasso, Klaus

    2015-01-01

    AIM: Seminal plasma offer a more organ-specific matrix for markers in prostatic disease. We hypothesized that C-type natriuretic peptide (CNP) expression may constitute such a new target. METHODS: Patients with benign prostatic hyperplasia, clinically localized and metastatic prostate cancer were...... examined for CNP and CNP precursor (proCNP) concentrations in blood and seminal plasma. Furthermore, CNP and the CNP receptor (NPR-B) mRNA contents in tissue from prostate and seminal vesicles were analyzed by qPCR. RESULTS: CNP and NPR-B concentrations decreased with increasing tumor burden (p = 0.......0027 and p = 0.0096, respectively). In contrast, seminal plasma CNP and proCNP concentrations were markedly increased with increased tumor burden (p prostate cancer....

  7. C-terminal peptide extension via gas-phase ion/ion reactions

    Science.gov (United States)

    Peng, Zhou; McLuckey, Scott A.

    2015-01-01

    The formation of peptide bonds is of great importance from both a biological standpoint and in routine organic synthesis. Recent work from our group demonstrated the synthesis of peptides in the gas-phase via ion/ion reactions with sulfo-NHS reagents, which resulted in conjugation of individual amino acids or small peptides to the N-terminus of an existing ‘anchor’ peptide. Here, we demonstrate a complementary approach resulting in the C-terminal extension of peptides. Individual amino acids or short peptides can be prepared as reagents by incorporating gas phase-labile protecting groups to the reactive C-terminus and then converting the N-terminal amino groups to the active ketenimine reagent. Gas-phase ion/ion reactions between the anionic reagents and doubly protonated “anchor” peptide cations results in extension of the “anchor” peptide with new amide bond formation at the C-terminus. We have demonstrated that ion/ion reactions can be used as a fast, controlled, and efficient means for C-terminal peptide extension in the gas phase. PMID:26640400

  8. The Association between Newborn Regional Body Composition and Cord Blood Concentrations of C-Peptide and Insulin-Like Growth Factor I.

    Directory of Open Access Journals (Sweden)

    Emma M Carlsen

    Full Text Available Third trimester fetal growth is partially regulated by C-peptide and insulin-like growth factor I (IGF-I. Prenatal exposures including maternal obesity and high gestational weight gain as well as high birth weight have been linked to subsequent metabolic disease. We evaluated the associations between newborn regional body composition and cord blood levels of C-peptide and IGF-I.We prospectively included obese and normal-weight mothers and their newborns; cord blood was collected and frozen. Analyses of C-peptide and IGF-I were performed simultaneously, after recruitment was completed. Newborn regional body composition was assessed with dual-energy X-ray absorptiometry scanning (DXA within 48 hours of birth.Three hundred thirty-six term infants were eligible to participate in the study; of whom 174 (52% infants had cord blood taken. Total, abdominal and arm and leg fat mass were positively associated with C-peptide (p < 0.001. Arm and leg fat mass was associated with IGF-I concentration: 28 g [95% confidence interval: 4, 53] per doubling of IGF-I. There was no association between total or abdominal fat mass and IGF-I. Fat-free mass was positively associated with both C-peptide (p < 0.001 and IGF-I (p = 0.004.Peripheral fat tissue accumulation was associated with cord blood C-peptide and IGF-I. Total and abdominal fat masses were related to C-peptide but not to IGF-I. Thus, newborn adiposity is partially mediated through C-peptide and early linear growth is associated with IGF-I.

  9. Saliva transit in patients with gastroesophageal reflux disease.

    Science.gov (United States)

    Cassiani, R A; Mota, G A; Aprile, L R O; Dantas, R O

    2015-10-01

    Saliva is an important factor in the neutralization of the acidity of the refluxed material that comes from the stomach to the esophagus. The impairment of saliva transit from oral cavity to distal esophagus may be one of the causes of esophagitis and symptoms in gastroesophageal reflux disease (GERD). With the scintigraphic method, the transit of 2 mL of artificial saliva was measured in 30 patients with GERD and 26 controls. The patients with GERD had symptoms of heartburn and acid regurgitation, a 24-hour pH monitoring with more than 4.2% of the time with pH below four, 26 with erosive esophagitis, and four with non-erosive reflux disease. Fourteen had mild dysphagia for solid foods. Twenty-one patients had normal esophageal manometry, and nine had ineffective esophageal motility. They were 15 men and 15 women, aged 21-61 years, mean 39 years. The control group had 14 men and 12 women, aged 19-61 years, mean 35 years. The subjects swallowed in the sitting and supine position 2 mL of artificial saliva labeled with 18 MBq of (99m) Technetium phytate. The time of saliva transit was measured from oral cavity to esophageal-gastric transition, from proximal esophagus to esophageal-gastric transition, and the transit through proximal, middle, and distal esophageal body. There was no difference between patients and controls in the time for saliva to go from oral cavity to esophageal-gastric transition, and from proximal esophagus to esophageal-gastric transition, in the sitting and supine positions. In distal esophagus in the sitting position, the saliva transit duration was shorter in patients with GERD (3.0 ± 0.8 seconds) than in controls (7.6 ± 1.7 seconds, P = 0.03). In conclusion, the saliva transit from oral cavity to the esophageal-gastric transition in patients with GERD has the same duration than in controls. Saliva transit through the distal esophageal body is faster in patients with GERD than controls. © 2014 International Society for Diseases of the

  10. C-terminal substitution of MDM2 interacting peptides modulates binding affinity by distinctive mechanisms.

    Directory of Open Access Journals (Sweden)

    Christopher J Brown

    Full Text Available The complex between the proteins MDM2 and p53 is a promising drug target for cancer therapy. The residues 19-26 of p53 have been biochemically and structurally demonstrated to be a most critical region to maintain the association of MDM2 and p53. Variation of the amino acid sequence in this range obviously alters the binding affinity. Surprisingly, suitable substitutions contiguous to this region of the p53 peptides can yield tightly binding peptides. The peptide variants may differ by a single residue that vary little in their structural conformations and yet are characterized by large differences in their binding affinities. In this study a systematic analysis into the role of single C-terminal mutations of a 12 residue fragment of the p53 transactivation domain (TD and an equivalent phage optimized peptide (12/1 were undertaken to elucidate their mechanistic and thermodynamic differences in interacting with the N-terminal of MDM2. The experimental results together with atomistically detailed dynamics simulations provide insight into the principles that govern peptide design protocols with regard to protein-protein interactions and peptidomimetic design.

  11. Pathological consequences of C-peptide deficiency ininsulin-dependent diabetes mellitus

    Institute of Scientific and Technical Information of China (English)

    Ahmad Ghorbani; Reza Shafiee-Nick

    2015-01-01

    Diabetes is associated with several complicationssuch as retinopathy, nephropathy, neuropathy andcardiovascular diseases. Currently, insulin is the mainused medication for management of insulin-dependentdiabetes mellitus (type-1 diabetes). In this metabolicsyndrome, in addition to decrease of endogenous insulin,the plasma level of connecting peptideC-peptide) is alsoreduced due to beta cell destruction. Studies in the pastdecade have shown that C-peptide is much more than abyproduct of insulin biosynthesis and possess differentbiological activities. Therefore, it may be possible thatC-peptide deficiency be involved, at least in part, in thedevelopment of different complications of diabetes. It hasbeen shown that a small level of remaining C-peptide isassociated with significant metabolic benefit. The purposeof this review is to describe beneficial effects of C-peptidereplacement on pathological features associated withinsulin-dependent diabetes. Also, experimental andclinical findings on the effects of C-peptide on wholebodyglucose utilization, adipose tissue metabolism andtissues blood flow are summarized and discussed. Thehypoglycemic, antilipolytic and vasodilator effects ofC-peptide suggest that it may contribute to fine-tuningof the tissues metabolism under different physiologic orpathologic conditions. Therefore, C-peptide replacementtogether with the classic insulin therapy may prevent,retard, or ameliorate diabetic complications in patientswith type-1 diabetes.

  12. Describing the Peptide Binding Specificity of HLA-C

    DEFF Research Database (Denmark)

    Rasmussen, Michael; Harndahl, Mikkel Nors; Nielsen, Morten

    for 5 HLA-C molecules and for all, but one, molecule we find a high frequency of binders, >70%, among these peptides. To extend the examined peptide space, we use bioinformatic prediction tools to search for additional binders. Finally, we update our prediction tool, NetMHCpan, with the HLA-C affinity......Human leukocyte antigen (HLA) presents peptides to T-cells for immune scrutiny. Whereas HLA-A and -B have been described in great detail, HLA-C has received much less attention. Here, to increase the coverage of HLA-C and the accuracy of the corresponding tools, we have generated HLA-C molecules...... data and show that the predictive performance for HLA-C molecules now is increased to a level comparable withthat of HLA-A and -B. These novel HLA-C molecules and predictors are successfully used to generate HLA-C tetramers and validate HLA-C-restricted T cell responses....

  13. Synthesis of peptide thioacids at neutral pH using bis(2-sulfanylethyl)amido peptide precursors.

    Science.gov (United States)

    Pira, Silvain L; Boll, Emmanuelle; Melnyk, Oleg

    2013-10-18

    Reaction of bis(2-sulfanylethyl)amido (SEA) peptides with triisopropylsilylthiol in water at neutral pH yields peptide thiocarboxylates. An alkylthioester derived from β-alanine was used to trap the released bis(2-sulfanylethyl)amine and displace the equilibrium toward the peptide thiocarboxylate.

  14. High Level Expression and Purification of the Clinically Active Antimicrobial Peptide P-113 in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Kuang-Ting Cheng

    2018-03-01

    Full Text Available P-113, which was originally derived from the human saliva protein histatin 5, is a histidine-rich antimicrobial peptide with the sequence AKRHHGYKRKFH. P-113 is currently undergoing phase II clinical trial as a pharmaceutical agent to fight against fungal infections in HIV patients with oral candidiasis. Previously, we developed a new procedure for the high-yield expression and purification of hG31P, an analogue and antagonist of human CXCL8. Moreover, we have successfully removed lipopolysaccharide (LPS, endotoxin associated with hG31P in the expression with Escherichia coli. In this paper, we have used hG31P as a novel fusion protein for the expression and purification of P-113. The purity of the expressed P-113 is more than 95% and the yield is 4 mg P-113 per liter of E. coli cell culture in Luria-Bertani (LB medium. The antimicrobial activity of the purified P-113 was tested. Furthermore, we used circular dichroism (CD and nuclear magnetic resonance (NMR spectroscopy to study the structural properties of P-113. Our results indicate that using hG31P as a fusion protein to obtain large quantities of P-113 is feasible and is easy to scale up for commercial production. An effective way of producing enough P-113 for future clinical studies is evident in this study.

  15. Patients with dental calculus have increased saliva and gingival crevicular fluid fetuin-A levels but no association with fetuin-A polymorphisms.

    Science.gov (United States)

    Doğan, Gülnihal Emrem; Demir, Turgut; Laloğlu, Esra; Sağlam, Ebru; Aksoy, Hülya; Yildirim, Abdulkadir; Akçay, Fatih

    2016-12-22

    Fetuin-A is a potent inhibitor of calcium-phosphate precipitation and of the calcification process, therefore it can also be related with dental calculus. Thus, we aimed to investigate a possible relationship between fetuin-A gene polymorphism and the presence of dental calculus. A possible relationship between serum, saliva and gingival crevicular fluid (GCF) levels of fetuin-A was also investigated. Fetuin-A c.742C > T and c.766C > G polymorphisms were investigated in 103 patients with or without dental calculus. Additionally, serum, saliva and GCF fetuin-A levels of patients were compared according to dental calculus presence. A significant difference was not observed in the distribution of the fetuin-A c.742C > T and c.766C > G polymorphisms between patients with or without dental calculus. Saliva and GCF fetuin-A concentrations of patients with dental calculus were statistically higher than those without dental calculus (P=0.001, P=0.036 respectively). According to our results, fetuin-A c.742C > T and c.766C > G polymorphisms were not associated with presence of dental calculus. However, higher GCF and saliva fetuin-A levels were detected in patients with dental calculus than in patients without dental calculus, which may result from an adaptive mechanism to inhibit mineral precipitation and eventually calculus formation.

  16. Patients with dental calculus have increased saliva and gingival crevicular fluid fetuin-A levels but no association with fetuin-A polymorphisms

    Directory of Open Access Journals (Sweden)

    Gülnihal Emrem DOĞAN

    Full Text Available ABSTRACT: Fetuin-A is a potent inhibitor of calcium-phosphate precipitation and of the calcification process, therefore it can also be related with dental calculus. Thus, we aimed to investigate a possible relationship between fetuin-A gene polymorphism and the presence of dental calculus. A possible relationship between serum, saliva and gingival crevicular fluid (GCF levels of fetuin-A was also investigated. Fetuin-A c.742C > T and c.766C > G polymorphisms were investigated in 103 patients with or without dental calculus. Additionally, serum, saliva and GCF fetuin-A levels of patients were compared according to dental calculus presence. A significant difference was not observed in the distribution of the fetuin-A c.742C > T and c.766C > G polymorphisms between patients with or without dental calculus. Saliva and GCF fetuin-A concentrations of patients with dental calculus were statistically higher than those without dental calculus (P=0.001, P=0.036 respectively. According to our results, fetuin-A c.742C > T and c.766C > G polymorphisms were not associated with presence of dental calculus. However, higher GCF and saliva fetuin-A levels were detected in patients with dental calculus than in patients without dental calculus, which may result from an adaptive mechanism to inhibit mineral precipitation and eventually calculus formation.

  17. C-di-GMP regulates antimicrobial peptide resistance in Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Chua, Song Lin; Tan, Sean Yang-Yi; Rybtke, Morten Theil

    2013-01-01

    Bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) is an intracellular second messenger which controls the life styles of many bacteria. A high intracellular level of c-di-GMP induces a biofilm lifestyle, whereas a low intracellular level of c-di-GMP stimulates dispersal of biofilms and promotes...... a planktonic lifestyle. Here, we used expression of different reporters to show that planktonic cells (PCells), biofilm cells (BCells) and cells dispersed from biofilms (DCells) had distinct intracellular c-di-GMP levels. Proteomics analysis showed that the low intracellular c-di-GMP level of DCells induced...... the expression of proteins required for the virulence and development of antimicrobial peptide resistance in P. aeruginosa. In accordance, P. aeruginosa cells with low c-di-GMP levels were found to be more resistant to colistin than P. aeruginosa cells with high c-di-GMP levels. This contradicts the current...

  18. Longitudinal Study of Hepatitis A Infection by Saliva Sampling: The Kinetics of HAV Markers in Saliva Revealed the Application of Saliva Tests for Hepatitis A Study

    Science.gov (United States)

    Amado Leon, Luciane Almeida; de Almeida, Adilson José; de Paula, Vanessa Salete; Tourinho, Renata Santos; Villela, Daniel Antunes Maciel; Gaspar, Ana Maria Coimbra; Lewis-Ximenez, Lia Laura; Pinto, Marcelo Alves

    2015-01-01

    Despite the increasing numbers of studies investigating hepatitis A diagnostic through saliva, the frequency and the pattern of hepatitis A virus (HAV) markers in this fluid still remains unknown. To address this issue, we carried on a longitudinal study to examine the kinetics of HAV markers in saliva, in comparison with serum samples. The present study followed-up ten patients with acute hepatitis A infection during 180 days post diagnosis (dpd). Total anti-HAV was detected in paired serum and saliva samples until the end of the follow-up, showing a peak titer at 90th. However, total anti-HAV level was higher in serum than in saliva samples. This HAV marker showed a probability of 100% to be detected in both serum and saliva during 180 dpd. The IgM anti-HAV could be detected in saliva up to 150 dpd, showing the highest frequency at 30th, when it was detected in all individuals. During the first month of HAV infection, this acute HAV marker showed a detection probability of 100% in paired samples. The detection of IgM anti-HAV in saliva was not dependent on its level in serum, HAV-RNA detection and/or viral load, since no association was found between IgM anti-HAV positivity in saliva and any of these parameter (p>0.05). Most of the patients (80%) were found to contain HAV-RNA in saliva, mainly at early acute phase (30th day). However, it was possible to demonstrate the HAV RNA presence in paired samples for more than 90 days, even after seroconversion. No significant relationship was observed between salivary HAV-RNA positivity and serum viral load, demonstrating that serum viral load is not predictive of HAV-RNA detection in saliva. Similar viral load was seen in paired samples (on average 104 copies/mL). These data demonstrate that the best diagnostic coverage can be achieved by salivary anti-HAV antibodies and HAV-RNA tests during 30–90 dpd. The long detection and high probability of specific-HAV antibodies positivity in saliva samples make the assessment of

  19. The relationship between MMPs and pH in whole saliva of radiated head and neck cancer patients.

    Science.gov (United States)

    Vuotila, T; Ylikontiola, L; Sorsa, T; Luoto, H; Hanemaaijer, R; Salo, T; Tjäderhane, L

    2002-07-01

    Radiation therapy for head and neck tumour patients may lead to decreased salivary flow, oral mucosal lesions and increased caries experience. Salivary matrix metalloproteinases (MMPs) may participate in the pathogenesis of mucosal lesions and dentinal caries. The aims of this study were: (i) to assay the presence, molecular forms and proteolytic activity of MMP-8 (collagenase-2) and MMP-9 (gelatinase B) in the whole saliva of head and neck cancer patients having radiation therapy; (ii) to see whether salivary pH affects the activity of MMP-8 and MMP-9; and (iii) to find out the possible connection between MMP-8 and MMP-9 with the eruption of oral mucosal lesions during radiation therapy. The whole saliva samples of 39 head and neck cancer patients having radiation therapy were collected before, during and after radiation therapy, and saliva flow rate, pH, buffer capacity, Streptococcus mutans, Lactobacillus and Candida albicans were measured. Any oral mucosal lesions were examined during each visit. The levels of MMP-8 were measured by immunofluorometric assay (IFMA) and the presence of different MMP-8 forms was analysed using Western immunoblotting. The presence and molecular forms of MMP-9 were analyzed by gelatin zymography. MMP-9 capture activity assay was used to determine the APMA-activated MMP-9 activity (total) and the endogenously active MMP-9 (free activity). Salivary flow rate, buffer capacity and pH decreased, and the levels of Lactobacilli increased significantly, during the first half of the radiation therapy. The endogenously activated salivary MMP-9 correlated with low salivary pH (P = 0.013). No connection was found between the oral mucosal lesions and salivary MMP-8 or MMP-9. In this study, salivary MMP-8 or MMP-9 did not correlate with the presence of radiation induced oral mucosal lesions, but the activation of MMP-9 may be dependent on pH.

  20. An evaluation of saliva as an alternative to plasma for the detection of hepatitis C virus antibodies

    Directory of Open Access Journals (Sweden)

    Moorthy M

    2008-01-01

    Full Text Available Purpose: Seroepidemiological studies on the prevalence of Hepatitis C virus (HCV in India have been hampered by reluctance of subjects to provide blood samples for testing. We evaluated the use of saliva as an alternate specimen to blood for the detection of antibodies to HCV. Methods: Chronic liver disease (CLD patients attending the liver clinic were recruited for this study. A saliva and plasma sample (sample pair was collected from each patient included in the study. Saliva samples were collected using a commercially available collection device - OmniSal. Sample pairs were tested with an in-use ELISA for the detection of antibodies to HCV (HCV-Ab, with a minor modification in the manufacturer′s protocol while testing saliva. The cut-off absorbance value for declaring a sample as positive was determined by receiver operating curve (ROC analysis. HCV-Ab positivity in saliva was compared with that in plasma as well as with viral load in plasma and infecting genotype of the virus. Sensitivity, specificity, positive and negative predictive values, and correlation coefficients were calculated using Medcalc statistical software. Results: The optimal accuracy indices were: sensitivity-81.6%; specificity-92.5%; PPV-85.1% and NPV-90.5%. No correlation was found between salivary positivity and HCV viral load in plasma or infecting genotype. Conclusions: The accuracy indices indicate that the assay must be optimized further before it can be recommended for routine use in epidemiological surveys for HCV-Ab.

  1. Prediction of Impending Type 1 Diabetes through Automated Dual-Label Measurement of Proinsulin:C-Peptide Ratio.

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    Annelien Van Dalem

    Full Text Available The hyperglycemic clamp test, the gold standard of beta cell function, predicts impending type 1 diabetes in islet autoantibody-positive individuals, but the latter may benefit from less invasive function tests such as the proinsulin:C-peptide ratio (PI:C. The present study aims to optimize precision of PI:C measurements by automating a dual-label trefoil-type time-resolved fluorescence immunoassay (TT-TRFIA, and to compare its diagnostic performance for predicting type 1 diabetes with that of clamp-derived C-peptide release.Between-day imprecision (n = 20 and split-sample analysis (n = 95 were used to compare TT-TRFIA (AutoDelfia, Perkin-Elmer with separate methods for proinsulin (in-house TRFIA and C-peptide (Elecsys, Roche. High-risk multiple autoantibody-positive first-degree relatives (n = 49; age 5-39 were tested for fasting PI:C, HOMA2-IR and hyperglycemic clamp and followed for 20-57 months (interquartile range.TT-TRFIA values for proinsulin, C-peptide and PI:C correlated significantly (r2 = 0.96-0.99; P<0.001 with results obtained with separate methods. TT-TRFIA achieved better between-day %CV for PI:C at three different levels (4.5-7.1 vs 6.7-9.5 for separate methods. In high-risk relatives fasting PI:C was significantly and inversely correlated (rs = -0.596; P<0.001 with first-phase C-peptide release during clamp (also with second phase release, only available for age 12-39 years; n = 31, but only after normalization for HOMA2-IR. In ROC- and Cox regression analysis, HOMA2-IR-corrected PI:C predicted 2-year progression to diabetes equally well as clamp-derived C-peptide release.The reproducibility of PI:C benefits from the automated simultaneous determination of both hormones. HOMA2-IR-corrected PI:C may serve as a minimally invasive alternative to the more tedious hyperglycemic clamp test.

  2. Pengaruh Berkumur Air Kelapa Muda Terhadap Ph Saliva

    OpenAIRE

    Mokoginta, Zuthra P

    2017-01-01

    PENGARUH BERKUMUR AIR KELAPA MUDA TERHADAP pH SALIVA Zuthra P. Mokoginta1) , Vonny N.S. Wowor1) , Juliatri1) 1)Program Studi Pendidikan Dokter Gigi, Fakultas Kedokteran UNSRAT Manado, 95115 ABSTRACT Saliva is one factor that contributes to the development of caries, particularly in the process of demineralization. The low pH in the oral cavity will facilitate the growth of acidogenic bacteria such as Streptococcus mutans and Lactobacillus which is the main cause microorganisms in th...

  3. Dendroaspis natriuretic peptide binds to the natriuretic peptide clearance receptor

    International Nuclear Information System (INIS)

    Johns, Douglas G.; Ao, Zhaohui; Heidrich, Bradley J.; Hunsberger, Gerald E.; Graham, Taylor; Payne, Lisa; Elshourbagy, Nabil; Lu, Quinn; Aiyar, Nambi; Douglas, Stephen A.

    2007-01-01

    Dendroaspis natriuretic peptide (DNP) is a newly-described natriuretic peptide which lowers blood pressure via vasodilation. The natriuretic peptide clearance receptor (NPR-C) removes natriuretic peptides from the circulation, but whether DNP interacts with human NPR-C directly is unknown. The purpose of this study was to test the hypothesis that DNP binds to NPR-C. ANP, BNP, CNP, and the NPR-C ligands AP-811 and cANP(4-23) displaced [ 125 I]-ANP from NPR-C with pM-to-nM K i values. DNP displaced [ 125 I]-ANP from NPR-C with nM potency, which represents the first direct demonstration of binding of DNP to human NPR-C. DNP showed high pM affinity for the GC-A receptor and no affinity for GC-B (K i > 1000 nM). DNP was nearly 10-fold more potent than ANP at stimulating cGMP production in GC-A expressing cells. Blockade of NPR-C might represent a novel therapeutic approach in augmenting the known beneficial actions of DNP in cardiovascular diseases such as hypertension and heart failure

  4. Rapid determination of natural steroidal hormones in saliva for the clinical diagnoses

    Directory of Open Access Journals (Sweden)

    Oh Jin-Aa

    2012-03-01

    Full Text Available Abstract Background Saliva samples are easily collectable and non-invasive, and the monitoring of natural steroidal hormones, such as estrone (E1, 17β-estradiol (E2, estriol (E3, progesterone (P, and testosterone (T, in saliva has attracted much attention due to its numerous potential clinical and health-related applications. Because E1, E2, E3, P and T are useful indicators in numerous clinical and health-related diagnoses, there is a need for simultaneous determination. Results A gas chromatography-mass spectrometric assay was developed for rapid simultaneous determination of E1, E2, E3, P and T in saliva for clinical diagnoses. Extraction was achieved with a liquid extraction using 3.0 mL of pentane. The extract was dried and silylated with N-methyl-N-(trimethylsilyl trifluoroacetamide/NH4I (100:2 under a catalysis of 1.5% dithioerythritol for 10 min at 90°C. The accuracy of the analytes was in the range of 96% to 112% at concentrations of 0.05 and 0.10 μg/L (5.0 and 10.0 μg/L for E3, respectively, with relative standard deviations of less than 11%. The lowest quantification limits were from 0.002 to 0.6 μg/L for 1.0 mL of saliva. Conclusion Natural steroidal hormones were detected in the concentration ranges of nd to 0.2 μg/L in human saliva. The salivary testosterone values in the patients with prostatic carcinoma were significantly lower than in normal males. The method may useful in numerous clinical and health-related diagnoses.

  5. Clinical trial participant characteristics and saliva and DNA metrics

    Directory of Open Access Journals (Sweden)

    Richards Julie

    2009-10-01

    Full Text Available Abstract Background Clinical trial and epidemiological studies need high quality biospecimens from a representative sample of participants to investigate genetic influences on treatment response and disease. Obtaining blood biospecimens presents logistical and financial challenges. As a result, saliva biospecimen collection is becoming more frequent because of the ease of collection and lower cost. This article describes an assessment of saliva biospecimen samples collected through the mail, trial participant demographic and behavioral characteristics, and their association with saliva and DNA quantity and quality. Methods Saliva biospecimens were collected using the Oragene® DNA Self-Collection Kits from participants in a National Cancer Institute funded smoking cessation trial. Saliva biospecimens from 565 individuals were visually inspected for clarity prior to and after DNA extraction. DNA samples were then quantified by UV absorbance, PicoGreen®, and qPCR. Genotyping was performed on 11 SNPs using TaqMan® SNP assays and two VNTR assays. Univariate, correlation, and analysis of variance analyses were conducted to observe the relationship between saliva sample and participant characteristics. Results The biospecimen kit return rate was 58.5% among those invited to participate (n = 967 and 47.1% among all possible COMPASS participants (n = 1202. Significant gender differences were observed with males providing larger saliva volume (4.7 vs. 4.5 ml, p = 0.019, samples that were more likely to be judged as cloudy (39.5% vs. 24.9%, p 0.21, P Conclusion Findings from this study show that demographic and behavioral characteristics of smoking cessation trial participants have significant associations with saliva and DNA metrics, but not with the performance of TaqMan® SNP or VNTR genotyping assays. Trial registration COMPASS; registered as NCT00301145 at clinicaltrials.gov.

  6. Relation of muscle indices with metabolic parameters and C-peptide in type 2 diabetes mellitus

    International Nuclear Information System (INIS)

    Tuzun, S.; Oner, C.; Dabak, M.R.; Kasikci, H.O.; Sargin, M.

    2017-01-01

    Objective: To assess the relation between bioimpedance measurements and metabolic parameters and C-peptide in patient with type 2 diabetes mellitus (DM). Study Design: Cross-sectional study. Place and Duration of Study: Kartal Dr Lutfi Kirdar Training and Research Hospital, Pendik Kaynarca Diabetes Center, Exercise and Metabolism Unit, between January and March 2015. Methodology: Patients with DM, aged less than 65 years, were assessed for bioimpedance analysis, fasting plasma glucose (FPG), HbA1c, C-peptide levels, triglyceride levels, LDL-cholesterol, and HDL-cholesterol levels. Skeletal muscle index, total muscle index, skeletal muscle percentage, and total muscle percentage were used for muscle-related analyses. Mann-Whitney U-test or independent t-test were used to compare differences between two independent groups. Pearson correlation test or Spearman correlation test were used to find out correlation between variables. Results: A total of 359 DM patients were enrolled in the study. Mean age was 51.6+-8.0 years, and 278 (77.7%) of the participants were females. After adjusting age and gender variables, there was no relation between muscle-related measurements and FPG, triglyceride, LDL-cholesterol (p>0.05). However, there was muscle-related indexes (MRI) positively correlation with C-peptide and inversely associated with HDL-cholesterol (p<0.05). Conclusion: Muscle-related indices positively correlated with C-peptide, which showed endogenous insulin reserve. (author)

  7. Novel tetra-peptide insertion in Gag-p6 ALIX-binding motif in HIV-1 subtype C associated with protease inhibitor failure in Indian patients.

    Science.gov (United States)

    Neogi, Ujjwal; Rao, Shwetha D; Bontell, Irene; Verheyen, Jens; Rao, Vasudev R; Gore, Sagar C; Soni, Neelesh; Shet, Anita; Schülter, Eugen; Ekstrand, Maria L; Wondwossen, Amogne; Kaiser, Rolf; Madhusudhan, Mallur S; Prasad, Vinayaka R; Sonnerborg, Anders

    2014-09-24

    A novel tetra-peptide insertion was identified in Gag-p6 ALIX-binding region, which appeared in protease inhibitor failure Indian HIV-1C sequences (odds ratio=17.1, P < 0.001) but was naturally present in half of untreated Ethiopian HIV-1C sequences. The insertion is predicted to restore ALIX-mediated virus release pathway, which is lacking in HIV-1C. The clinical importance of the insertion needs to be evaluated in HIV-1C dominating regions wherein the use of protease inhibitor drugs are being scaled up.

  8. Efficacy of a novel antimicrobial peptide against periodontal pathogens in both planktonic and polymicrobial biofilm states.

    Science.gov (United States)

    Wang, Hong-Yan; Cheng, Jya-Wei; Yu, Hui-Yuan; Lin, Li; Chih, Ya-Han; Pan, Ya-Ping

    2015-10-01

    Streptococcus gordonii, Fusobacterium nucleatum and Porphyromonas gingivalis represent the early, middle and late colonizers of the bacterial accretion in dental plaque biofilms. These sessile communities constitute a protected mode of growth that promotes survival in a hostile environment. This study describes a novel and unrecognized role for a synthetic cationic antimicrobial peptide, Nal-P-113, which inhibits and kills periodontal bacteria in planktonic state, inhibits the formation of biofilms and eradicates polymicrobial biofilms. Nal-P-113 is also stable in saliva, serum and saline solution. At a concentration less than 320 μg/mL which is harmless to normal oral cells, Nal-P-113 can kill bacteria in planktonic state. At a concentration of antimicrobial peptide Nal-P-113 (1280 μg/mL) which only causes slight damages to normal oral cells is needed to kill bacteria in biofilm state. It is worth mentioning that this concentration of Nal-P-113 is harmless to rat oral mucosa compared to chlorhexidine. The mechanism of Nal-P-113 inhibiting and killing periodontal bacteria might rely on the abilities to permeabilize and/or to form pores within the cytoplasmic membranes, thus causes the death of bacteria. Here, we provided a novel and stable antimicrobial peptide with very low mammalian cytotoxicity, which can inhibit and kill periodontal bacteria in both planktonic and polymicrobial biofilm states. Nal-P-113 is a potent antimicrobial peptide with strong antimicrobial ability, improved deficiency compared with other antibacterial peptides, and remains stable in phosphate buffered saline, saliva, brain-heart infusion medium and bovine calf serum. Nal-P-113 exhibits a broad spectrum of bacteriocidal activity with excellent eradicating capability on oral pathogens and the respective biofilms. In this study, we used propidium iodide staining, scanning electron microscopy and transmission electron microscopy to confirm that Nal-P-113 can perforate plasmalemma thereby

  9. Comparative evaluation of qigong on various parameters of saliva

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    Bayat Movahed S.

    2007-05-01

    Full Text Available Background and Aim: Qigong is a type of Chinese psychosomatic exercise that integrates meditation, slow physical movements, and breathing. Numerous physical and mental benefits have been classically ascribed to qigong. On the other hand, unstimulated saliva is thought to play an important role in oral immunity, enamel stability and moisturizing the oral mucous membrane. Impaired salivary secretion leads to dental caries, mucosal deterioration and mouth dryness. The aim of the present study was to analyze the effect of a qigong program on various parameters of saliva such as quantity of unstimulated saliva, pH and SIgA. Materials and Methods: Twenty three subjects participated in this clinical trial study. The experimental subjects underwent a qigong training program, conducted by a qualified instructor. The program consisted of half an hour daily practice for 6 months (spring and winter. Saliva was collected in two periods: once during the spring before the experiment commencement and the second, in the following spring. During each period saliva collection was done on tuesday of each week. pH and quantity of salvia measurements were taken simultaneously. SIgA measurements were also taken based on the values obtained in the last phase of the experiment at the end of each spring. The results were analyzed using paired sample T test, one way repeated measure and Bon Ferroni multiple comparison. P<0.05 was the level of significance. Results: Based on our findings, the change in the amount of unstimulated salvia as well as SIgA was statistically significant (P<0.001; however, there was no significant difference in pH values before and after experiment. Conclusion: These findings demonstrate that after 6 months of practicing qigong, significant changes in amount of unstimulated saliva and SIgA occurred in participants. The authors suggest that qigong may be a beneficial adjunctive treatment that enhances amount of unstimulated saliva and SIgA.

  10. Determination of carbamazepine in serum and saliva samples by high performance liquid chromatography with ultraviolet detection.

    Science.gov (United States)

    Dordević, Snezana; Kilibarda, Vesna; Stojanović, Tomislav

    2009-05-01

    Carbamazepine is antiepileptic drug widely used for the treatment of epilepsy. Due to low therapeutic index of carbamazepine there is a need for routine measuring its concentrations in biological fluids. The aim of the study was to describe a method for concomitant determination of carbamazepine in the serum and saliva. Separation of the drug from matrix is achieved by reversed-phase chromatography on a C18 column, with a mobile phase of methanol-water-acetic acid (65:34:1) at a flow-rate of 1.0 ml/min. Detection was effected by ultra-violet absorption at 285 nm. The total run time was 5 min. Samples were prepared by alkaline extraction (pH 10) using chlorophorm. Calibration curves were in the range 0.1-5 microg/mL for serum and saliva samples. Mean recoveries of spiked serum and saliva were 97.59 and 92.30%, respectively. Limits of detection (LOD) of carbamazepine in serum and saliva were 0.166 and 0.178 microg/mL, respectively. Limits of quantification (LOQ) in the serum and saliva were 0.237 and 0.226 microg/mL, respectively. The method precision was carried out with coefficient of variation of 2.10% and 4.03% for the serum and saliva, respectively. The obtained data showed that there was a strong correlation between saliva and serum concentrations (r = 0.9481, p < 0.001). The method described here is rapid, precise, accurate and simple, and can be used for quantitative determination of carbamazepine in human serum and saliva after therapy applying. Saliva samples could be used as an alternative matrix for therapeutic drug monitoring of this antiepileptic drug.

  11. Development of SI-traceable C-peptide certified reference material NMIJ CRM 6901-a using isotope-dilution mass spectrometry-based amino acid analyses.

    Science.gov (United States)

    Kinumi, Tomoya; Goto, Mari; Eyama, Sakae; Kato, Megumi; Kasama, Takeshi; Takatsu, Akiko

    2012-07-01

    A certified reference material (CRM) is a higher-order calibration material used to enable a traceable analysis. This paper describes the development of a C-peptide CRM (NMIJ CRM 6901-a) by the National Metrology Institute of Japan using two independent methods for amino acid analysis based on isotope-dilution mass spectrometry. C-peptide is a 31-mer peptide that is utilized for the evaluation of β-cell function in the pancreas in clinical testing. This CRM is a lyophilized synthetic peptide having the human C-peptide sequence, and contains deamidated and pyroglutamylated forms of C-peptide. By adding water (1.00 ± 0.01) g into the vial containing the CRM, the C-peptide solution in 10 mM phosphate buffer saline (pH 6.6) is reconstituted. We assigned two certified values that represent the concentrations of total C-peptide (mixture of C-peptide, deamidated C-peptide, and pyroglutamylated C-peptide) and C-peptide. The certified concentration of total C-peptide was determined by two amino acid analyses using pre-column derivatization liquid chromatography-mass spectrometry and hydrophilic chromatography-mass spectrometry following acid hydrolysis. The certified concentration of C-peptide was determined by multiplying the concentration of total C-peptide by the ratio of the relative area of C-peptide to that of the total C-peptide measured by liquid chromatography. The certified value of C-peptide (80.7 ± 5.0) mg/L represents the concentration of the specific entity of C-peptide; on the other hand, the certified value of total C-peptide, (81.7 ± 5.1) mg/L can be used for analyses that does not differentiate deamidated and pyroglutamylated C-peptide from C-peptide itself, such as amino acid analyses and immunochemical assays.

  12. Conserved peptides within the E2 region of Hepatitis C virus induce humoral and cellular responses in goats

    Directory of Open Access Journals (Sweden)

    El Shenawy Reem

    2009-05-01

    Full Text Available Abstract The reason(s why human antibodies raised against hepatitis C virus (HCV E2 epitopes do not offer protection against multiple viral infections may be related to either genetic variations among viral strains particularly within the hypervariable region-1 (HVR-1, low titers of anti E2 antibodies or interference of non neutralizing antibodies with the function of neutralizing antibodies. This study was designed to assess the immunogenic properties of genetically conserved peptides derived from the C-terminal region of HVR-1 as potential therapeutic and/or prophylactic vaccines against HCV infection. Goats immunized with E2-conserved synthetic peptides termed p36 (a.a 430–446, p37(a.a 517–531 and p38 (a.a 412–419 generated high titers of anti-p36, anti-p37 and anti-P38 antibody responses of which only anti- p37 and anti- p38 were neutralizing to HCV particles in sera from patients infected predominantly with genotype 4a. On the other hand anti-p36 exhibited weak viral neutralization capacity on the same samples. Animals super-immunized with single epitopes generated 2 to 4.5 fold higher titers than similar antibodies produced in chronic HCV patients. Also the studied peptides elicited approximately 3 fold increase in cell proliferation of specific antibody-secreting peripheral blood mononuclear cells (PBMC from immunized goats. These results indicate that, besides E1 derived peptide p35 (a.a 315–323 described previously by this laboratory, E2 conserved peptides p37 and p38 represent essential components of a candidate peptide vaccine against HCV infection.

  13. Chitosan-Poly (I:C-PADRE Based Nanoparticles as Delivery Vehicles for Synthetic Peptide Vaccines

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    Jorge F. Correia-Pinto

    2015-09-01

    Full Text Available The safety and precision of peptide antigens has prompted the search for adjuvants capable of increasing the immune response against these intrinsically poorly immunogenic antigens. The integration of both immunostimulants and peptide antigens within nanometric delivery systems for their co-delivery to immune cells is a promising vaccination strategy. With this in mind, the potential synergistic effect of the immunostimulant poly (I:C (pIC and a T-Helper peptide (PADRE, integrated into a chitosan (CS based nanostructure, was explored. The value of this nanostructured combination of materials was assessed for a peptide antigen (1338aa derived from the HPV-16 L2 protein. These nanoparticles, produced by ionic gelation technique, exhibited a nanometric size (<300 nm, a high positive surface charge (>40 mV and high pIC association efficiency (>96%. They also showed capacity for the association of both the 1338aa and PADRE peptides. The influence of the presence of pIC and PADRE in the nanocomposition, as well as that of the peptide presentation form (encapsulated versus surface adsorbed on the antibody induction was evaluated in a preliminary in vivo study. The data obtained highlights the possibility to engineer nanoparticles through the rational combination of a number of adjuvant molecules together with the antigen.

  14. The association between saliva control, silent saliva penetration, aspiration, and videofluoroscopic findings in Parkinson's disease patients

    Science.gov (United States)

    Rajaei, Ali; Ashtari, Fereshteh; Azargoon, Seyed Abolfazl; Chitsaz, Ahmad; Nilforoush, Mohammad Hussein; Taheri, Masoud; Sadeghi, Saba

    2015-01-01

    Background: Dysphagia is a common disorder among patients with Parkinson's disease (PD). It occurs in up to 80% of all (PD) patients during the early stages of the disease and up to 95% in the advanced stages; but professionals may not hear from the patients about dysphagia symptoms until these symptoms reach an advanced stage and lead to medical complications. Materials and Methods: Thirty-three PD patients (mean age 66.09 ± 9.4 years; 24 men, nine women) participated in this study at our Neurology Institute, between April 20, 2013, and October 26, 2013. They were asked two questions; one about saliva control and the other about silent saliva penetration and aspiration. Next, they underwent the videofluoroscopic swallowing study (VFSS). Results: The Pearson Correlation coefficient between the Penetration–Aspiration Scale (PAS) scores and question 1 scores was 0.48 (P < 0.05, =0.25), and there was a significant correlation between the PAS scores and question 2 scores, and also question 1 scores + question 2 scores (r = 0.589, P < 0.05, =0 and r = 0589, P < 0.05, =0). Conclusions: This study showed a significant correlation between the questions about saliva control, silent saliva penetration, and aspiration, and laryngeal penetration and aspiration during VFSS. Therefore, by using these two questions, the potential silent laryngeal penetration and aspiration during meals could be detected before it led to aspiration pneumonia. Taking the benefit of these questions, as a part of the swallowing assessment of PD patients, is recommended. PMID:26261810

  15. Saliva from nymph and adult females of Haemaphysalis longicornis: a proteomic study.

    Science.gov (United States)

    Tirloni, Lucas; Islam, Mohammad Saiful; Kim, Tae Kwon; Diedrich, Jolene K; Yates, John R; Pinto, Antônio F M; Mulenga, Albert; You, Myung-Jo; Da Silva Vaz, Itabajara

    2015-06-24

    Haemaphysalis longicornis is a major vector of Theileria spp., Anaplasma phagocytophilum, Babesia spp. and Coxiella burnetti in East Asian countries. All life stages of ixodid ticks have a destructive pool-feeding style in which they create a pool-feeding site by lacerating host tissue and secreting a variety of biologically active compounds that allows the tick to evade host responses, enabling the uptake of a blood meal. The identification and functional characterization of tick saliva proteins can be useful to elucidate the molecular mechanisms involved in tick development and to conceive new anti-tick control methods. H. longicornis tick saliva was collected from fully engorged nymphs and fully engorged adults induced by dopamine or pilocarpine, respectively. Saliva was digested with trypsin for LC-MS/MS sequencing and peptides were searched against tick and rabbit sequences. A total of 275 proteins were identified, of which 135 were tick and 100 were rabbit proteins. Of the tick proteins, 30 proteins were identified exclusively in fully engorged nymph saliva, 74 in fully engorged adult females, and 31 were detected in both stages. The identified tick proteins include heme/iron metabolism-related proteins, oxidation/detoxification proteins, enzymes, proteinase inhibitors, tick-specific protein families, and cytoskeletal proteins. Proteins involved in signal transduction, transport and metabolism of carbohydrate, energy, nucleotide, amino acids and lipids were also detected. Of the rabbit proteins, 13 were present in nymph saliva, 48 in adult saliva, and 30 were present in both. The host proteins include immunoglobulins, complement system proteins, antimicrobial proteins, serum albumin, peroxiredoxin, serotransferrin, apolipoprotein, hemopexin, proteinase inhibitors, and hemoglobin/red blood cells-related products. This study allows the identification of H. longicornis saliva proteins. In spontaneously detached tick saliva various proteins were identified

  16. A Novel Soluble Peptide with pH-Responsive Membrane Insertion.

    Science.gov (United States)

    Nguyen, Vanessa P; Alves, Daiane S; Scott, Haden L; Davis, Forrest L; Barrera, Francisco N

    2015-11-03

    Several diseases, such as cancer, are characterized by acidification of the extracellular environment. Acidosis can be employed as a target to specifically direct therapies to the diseased tissue. We have used first principles to design an acidity-triggered rational membrane (ATRAM) peptide with high solubility in solution that is able to interact with lipid membranes in a pH-dependent fashion. Biophysical studies show that the ATRAM peptide binds to the surface of lipid membranes at pH 8.0. However, acidification leads to the peptide inserting into the lipid bilayer as a transmembrane α-helix. The insertion of ATRAM into membranes occurs at a moderately acidic pH (with a pK of 6.5), similar to the extracellular pH found in solid tumors. Studies with human cell lines showed a highly efficient pH-dependent membrane targeting, without causing toxicity. Here we show that it is possible to rationally design a soluble peptide that selectively targets cell membranes in acidic environments.

  17. Adrenal status assessed by direct radioimmunoassay of cortisol in whole saliva or parotid saliva

    International Nuclear Information System (INIS)

    Walker, R.F.; Riad-Fahmy, D.; Read, G.F.

    1978-01-01

    We describe a direct radioimmunoassay for cortisol in 10-μl volumes of parotid saliva or whole saliva. Binding proteins are absent from these fluids, as demonstrated by the excellent correlation between results for samples assayed directly and by a comparison procedure involving extraction with 1,2-dichloroethane. The direct assay is specific, precise, and had a lower limit of sensitivity of 4 pg per assay tube. Comparison of cortisol concentrations in plasma, parotid saliva, and whole saliva in persons undergoing investigations for assessing adrenal function, including stimulation with cosyntropin (Synachthen) and suppression with dexamethasone, indicated that changes in plasma cortisol concentration were accurately and immediately reflected in saliva from either the parotid-gland or whole saliva. A marked circadian rhythm has also been demonstrated for cortisol in parotid-gland saliva and whole saliva. We had to modify the 1,2-dichloroethane extraction procedure for accurate determination of cortisol in parotid saliva and whole saliva of patients undergoing treatment with metyrapone

  18. Microstructure and nanomechanical properties of enamel remineralized with asparagine–serine–serine peptide

    International Nuclear Information System (INIS)

    Chung, Hsiu-Ying; Li, Cheng Che

    2013-01-01

    A highly biocompatible peptide, triplet repeats of asparagine–serine–serine (3NSS) was designed to regulate mineral deposition from aqueous ions in saliva for the reconstruction of enamel lesions. Healthy human enamel was sectioned and acid demineralized to create lesions, then exposed to the 3NSS peptide solution, and finally immersed in artificial saliva for 24 h. The surface morphology and roughness were examined using scanning electron microscopy (SEM) and atomic force microscopy (AFM), respectively. X-ray diffraction (XRD) was used to identify the phases and crystallinity of the deposited minerals observed on the enamel surface. Attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) was used to quantitatively analyze the mineral variation by calculating the relative integrated-area of characteristic bands. Nanohardness and elastic modulus measured by nanoindentation at various treatment stages were utilized to evaluate the degree of recovery. Biomimetic effects were accessed according to the degree of nanohardness recovery and the amount of hydroxyapatite deposition. The charged segments in the 3NSS peptide greatly attracted aqueous ions from artificial saliva to form hydroxyapatite crystals to fill enamel caries, in particular the interrod areas, resulting in a slight reduction in overall surface roughness. Additionally, the deposited hydroxyapatites were of a small crystalline size in the presence of the 3NSS peptide, which effectively restrained the plastic deformations and thus resulted in greater improvements in nanohardness and elastic modulus. The degree of nanohardness recovery was 5 times greater for remineralized enamel samples treated with the 3NSS peptide compared to samples without peptide treatment. - Highlights: ► The degree of nanohardness recovery of enamel was 4 times greater with the aid of 3NSS peptide. ► 3NSS peptide promoted the formation of hydroxyapatites with a smaller crystalline size (14 nm). ► 3NSS

  19. C-peptide comparative radioimmunoassays: a study of three commercial kits

    International Nuclear Information System (INIS)

    Villaume, C.; Beck, B.

    1983-01-01

    Plasma C-peptide immunoreactivity (CPR) was measured in 18 fasting subjects with three different commercial kits (RIA-mat C-peptide-, Byk-Mallinckrodt; RIA-gnost-hC-peptide, Hoechst-Behring; human C-peptide radioimmunoassay kit, Novo) The subjects were chosen as to cover a wide range of CPR concentrations (five healthy subjects, six obese subjects, three insulin-dependent diabetics, four normal subjects whose plasmas had been kept at - 20 0 C for periods of 16 or 36 months). CPR was measured with the Novo kit in eight other plasmas which were kept over a period of 36 months, with or without aprotinin. Good correlations have been established among the values found with the three kits. However, absolute concentration values for each subject as well as the dispersion of all plasma C-peptide values varied as a function of the kit used because of antibody specificity differences and because of the various separation methods. The normal range proposed changes with each kit and the blood CPR of a subject can be a normal, reduced or increased one, depending on the kit used. After several months of storage, plasma CPR degradation is observed with the three kits. A protease-inhibitor is necessary in order to avoid this C-peptide degradation due to the apparent existence of a plasma proteolytic enzyme

  20. Effect of dental restorative materials on total antioxidant capacity and calcium concentration of unstimulated saliva.

    Science.gov (United States)

    Ramezani, Gholam H; Moghadam, Mona-Momeni; Saghiri, Mohammad-Ali; Garcia-Godoy, Franklin; Asatourian, Armen; Aminsobhani, Mohsen; Scarbecz, Mark; Sheibani, Nader

    2017-01-01

    To evaluate the effect of dental amalgam and composite restorations on total antioxidant capacity (TAC) and calcium (Ca) ion concentration of unstimulated saliva. Forty-eight children aged 6-10 years selected and divided into three groups of sixteen (8 males, 8 females). In group A and B, samples consisted of two class II dental composite or amalgam restorations, while in group C samples were caries-free (control group). Unstimulated saliva from all samples was collected and TAC was measured by spectrophotometry using an adaptation of 2, 2'-azino-di-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) assay. The Ca ion level was estimated by an auto- analyzer. Data were analyzed with one- and two-way ANOVA test, at a p difference between groups ( p differences within and between groups ( p Gender is an effective factor in changes induced in oral cavity as females showed more emphatic reaction to dental filling materials than males. Patients who have dental restorations, especially dental composites, should pay more attention to their dental hygiene, because dental restorations can increase oxidative stress and decrease Ca ion level in saliva, which might jeopardize remineralization process of tooth structures after demineralization. Key words: Amalgam, caries, composite, saliva, total antioxidant capacity.

  1. Identification of novel inhibitors of Pseudomonas aeruginosa MurC enzyme derived from phage-displayed peptide libraries.

    Science.gov (United States)

    El Zoeiby, Ahmed; Sanschagrin, François; Darveau, André; Brisson, Jean-Robert; Levesque, Roger C

    2003-03-01

    The machinery of peptidoglycan biosynthesis is an ideal site at which to look for novel antimicrobial targets. Phage display was used to develop novel peptide inhibitors for MurC, an essential enzyme involved in the early steps of biosynthesis of peptidoglycan monomer. We cloned and overexpressed the murA, -B and -C genes from Pseudomonas aeruginosa in the pET expression vector, adding a His-tag to their C termini. The three proteins were overproduced in Escherichia coli and purified to homogeneity in milligram quantities. MurA and -B were combinatorially used to synthesize the MurC substrate UDP-N-acetylmuramate, the identity of which was confirmed by mass spectrometry and nuclear magnetic resonance analysis. Two phage-display libraries were screened against MurC in order to identify peptide ligands to the enzyme. Three rounds of biopanning were carried out, successively increasing elution specificity from round 1 to 3. The third round was accomplished with both non-specific elution and competitive elution with each of the three MurC substrates, UDP-N-acetylmuramic acid (UNAM), ATP and L-alanine. The DNA of 10 phage, selected randomly from each group, was extracted and sequenced, and consensus peptide sequences were elucidated. Peptides were synthesized and tested for inhibition of the MurC-catalysed reaction, and two peptides were shown to be inhibitors of MurC activity with IC(50)s of 1.5 and 0.9 mM, respectively. The powerful selection technique of phage display allowed us to identify two peptide inhibitors of the essential bacterial enzyme MurC. The peptide sequences represent the basis for the synthesis of inhibitory peptidomimetic molecules.

  2. Peptide functionalized gold nanoparticles: the influence of pH on binding efficiency

    Science.gov (United States)

    Harrison, Emma; Hamilton, Jeremy W. J.; Macias-Montero, Manuel; Dixon, Dorian

    2017-07-01

    We report herein on the synthesis of mixed monolayer gold nanoparticles (AuNPs) capped with both polyethylene glycol (PEG) and one of three peptides. Either a receptor-mediated endocytosis peptide, an endosomal escape pathway (H5WYG) peptide or the Nrp-1 targeting RGD peptide (CRGDK) labeled with FITC. All three peptides have a thiol containing cysteine residue which can be used to bind the peptides to the AuNPs. In order to investigate the influence of pH on peptide attachment, PEGylated AuNPs were centrifuged, the supernatant removed, and the nanoparticles were then re-suspended in a range of pH buffer solutions above, below and at the respective isoelectric points of the peptides before co-functionalization. Peptide attachment was investigated using dynamic light scattering, Ultra-violet visible spectroscopy (UV/Vis), FTIR and photo luminescence spectroscopy. UV/Vis analysis coupled with protein assay results and photoluminescence of the FITC tagged RGD peptide concluded that a pH of ∼8 optimized the cysteine binding and stability, irrespective of the peptide used.

  3. Application of reaction type of C-peptide release test in diabetes mellitus

    International Nuclear Information System (INIS)

    Chen Dong; Duan Wenruo; He Juan; Lu Zhenfang

    2001-01-01

    The author is to confirm the effect of C-peptide release test and types of release reaction in appraisal of pancreas function of β-cell and selection of treatment for diabetes mellitus (DM) patients. The serum C-peptide release test of 67 normal controls and 217 DM patients were determined by RIA, and the results were analyzed and compared. C-peptide release test can reflect the pancreas function of β-cell better, the peak of C-peptide ≥ 0.6 nmol/L after lunch can be the limit of whether to reduce the level of blood glucose only by oral drug. The authors should adjust the treatment through analyzing the type of C-peptide release reaction. C-peptide release test is very important in evaluating the pancreas function of β-cell, classifying the type of DM and selecting the treatment

  4. Determination of carbamazepine in serum and saliva samples by high performance liquid chromatography with ultraviolet detection

    Directory of Open Access Journals (Sweden)

    Đorđević Snežana

    2009-01-01

    Full Text Available Background/Aim. Carbamazepine is antiepileptic drug widely used for the treatment of epilepsy. Due to low therapeutic index of carbamazepine there is a need for routine measuring its concentrations in biological fluids. The aim of the study was to describe a method for concomitant determination of carbamazepine in the serum and saliva. Methods. Separation of the drug from matrix is achieved by reversedphase chromatography on a C18 column, with a mobile phase of methanol-water-acetic acid (65:34:1 at a flow-rate of 1.0 ml/min. Detection was effected by ultra-violet absorption at 285 nm. The total run time was 5 min. Samples were prepared by alkaline extraction (pH 10 using chlorophorm. Results. Calibration curves were in the range 0.1-5 μg/mL for serum and saliva samples. Mean recoveries of spiked serum and saliva were 97.59 and 92.30%, respectively. Limits of detection (LOD of carbamazepine in serum and saliva were 0.166 and 0.178 μg/mL, respectively. Limits of quantification (LOQ in the serum and saliva were 0.237 and 0.226 μg/mL, respectively. The method precision was carried out with coefficient of variation of 2.10% and 4.03% for the serum and saliva, respectively. The obtained data showed that there was a strong correlation between saliva and serum concentrations (r = 0.9481, p < 0.001. Conclusion. The method described here is rapid, precise, accurate and simple, and can be used for quantitative determination of carbamazepine in human serum and saliva after therapy applying. Saliva samples could be used as an alternative matrix for therapeutic drug monitoring of this antiepileptic drug.

  5. The association between saliva control, silent saliva penetration, aspiration, and videofluoroscopic findings in Parkinson′s disease patients

    Directory of Open Access Journals (Sweden)

    Ali Rajaei

    2015-01-01

    Full Text Available Background: Dysphagia is a common disorder among patients with Parkinson′s disease (PD. It occurs in up to 80% of all (PD patients during the early stages of the disease and up to 95% in the advanced stages; but professionals may not hear from the patients about dysphagia symptoms until these symptoms reach an advanced stage and lead to medical complications. Materials and Methods: Thirty-three PD patients (mean age 66.09 ± 9.4 years; 24 men, nine women participated in this study at our Neurology Institute, between April 20, 2013, and October 26, 2013. They were asked two questions; one about saliva control and the other about silent saliva penetration and aspiration. Next, they underwent the videofluoroscopic swallowing study (VFSS. Results: The Pearson Correlation coefficient between the Penetration-Aspiration Scale (PAS scores and question 1 scores was 0.48 (P < 0.05, =0.25, and there was a significant correlation between the PAS scores and question 2 scores, and also question 1 scores + question 2 scores (r = 0.589, P < 0.05, =0 and r = 0589, P < 0.05, =0. Conclusions: This study showed a significant correlation between the questions about saliva control, silent saliva penetration, and aspiration, and laryngeal penetration and aspiration during VFSS. Therefore, by using these two questions, the potential silent laryngeal penetration and aspiration during meals could be detected before it led to aspiration pneumonia. Taking the benefit of these questions, as a part of the swallowing assessment of PD patients, is recommended.

  6. PERBEDAAN ANTARA pH SALIVA DAN AKTIVITAS ENZIM AMILASE MAHASISWA YANG MEROKOK DENGAN MAHASISWA YANG TIDAK MEROKOK

    OpenAIRE

    Syauqi, Ahmad; Humaryanto, Humaryanto

    2018-01-01

    ABSTRACT Introduction: Cigarettes can cause disturbances in the oral cavity. From previous research it is known that the daily consumption of cigarettes increases the decrease of salivary secretion and the content of bicarbonate. This will have an effect on the decrease of pH saliva. Low salivary pH also has an impact on decreased amylase enzyme activity. Departing from the basic theory, the researchers wanted to see if there are differences in salivary pH and amylase enzyme activity in st...

  7. [Correlation analysis of the pre-pregnant body mass index, the gestational weight gain and umbilical cord blood C peptide].

    Science.gov (United States)

    Chen, X L; Han, Y; Zhao, X M; Liu, Y; Lü, J J

    2017-01-03

    Objective: To explore the correlation among the pre-pregnant body mass index (BMI), gestational weight gain (GWG) and umbilical cord blood C peptide, and to investigate the influence of maternal weight management on the incidence of baby long-term metabolic syndrome. Methods: During May to Aug.2015, 485 pregnant women in Zhejiang Taizhou first people's hospital and Taizhou Huangyan maternal &child care service centre were selected in random and divided into four groups according to pre-pregnant BMI: low body mass, normal body mass, over body mass and obese group, and also divided into two groups for getting gestational diabetes mellitus (GDM) or not. According to the gestational weight gain (GWG), all the cases were divided into two groups: above the Institute Of Medicine (IMO) 2009 recommendations or not. According to the outcome, the GDM group which had received weight control treatment, was divided into successful treat group or not. At last, we tested the umbilical cord blood C peptide and birth weight of each newborn and compared the difference in all subgroups. The correlation between the umbilical cord blood C peptide and birth weight were analysed. Results: (1) In the pre-pregnant BMI groups, there were significant differences of incidence of GDM ( P 0.05). (2) Newborn birth weight and the umbilical cord blood C peptide were positively correlated ( r =0.673, P newborn birth weight in all subgroups. (4) All groups showed correlations with neonatal umbilical cord blood C peptide, the GDM mostly (β=0.58), pre-pregnant BMI secondly (β: 0.36, 0.38) , and GWG weakly (β=0.17). (5) By stratification analysis, in GDM group, low body mass was negatively correlated with umbilical cord blood C peptide ( P gestational weight gain and umbilical cord blood C peptide. Suitable maternal weight control especially pre-pregnant body mass index control will lower the baby long-term metabolic syndrome incidence.

  8. C-Peptides for diagnostics and therapy: a veterinary medicine point of view

    Directory of Open Access Journals (Sweden)

    Derek A. Rosenfield

    Full Text Available ABSTRACT: Empirical studies proved that C-peptides are performing numerous intrinsic biological roles, and serve as a marker for pancreatic performance analysis. Since the last decade, C-peptide assays for differential diagnosis in veterinary diabetic patients are becoming more available, but still only for a very limited number of species. Studies on C-peptide as a diagnostic tool, therapy for associated complications, or as replacement therapies for C-peptide deficiency still showed not to be a common practice in veterinary medicine. This review was conducted to determine the potential importance of C-peptide in Veterinary Medicine, relevant in the diagnosis of diabetes and for other metabolic processes, as well as its proposed therapeutic benefits. Numerous articles were identified that reported positive results in their experimental studies, whether C-peptide as a biomarker for pancreatic performance in dogs, cats, and horses, as a non-invasive method to monitor nutritional status in primates, or to investigate its potential therapeutic benefits for diabetes-related illnesses.

  9. Automatic computation of radioimmunoassay data. Insulin and C-peptide

    Energy Technology Data Exchange (ETDEWEB)

    Toyota, T; Kudo, M; Abe, K [Hirosaki Univ., Aomori (Japan). School of Medicine; Kawamata, F; Uehata, S

    1975-09-01

    Radioimmunoassay provided dose response curves which showed linearity by the use of logistic transformation (Rodbard). This transformation which was applicable to radioimmunoassay should be useful for the computer processing of insulin and C-peptide assay. In the present studies, standard curves were analysed by testing the fit of analytic functions to radioimmunoassay of insulin and C-peptides. A program for use in combination with the double antibody technique was made by Dr. Kawamata. This approach was evidenced to be useful in order to allow automatic computation of data derived from the double antibody assays of insulin and C-peptides. Automatic corrected calculations of radioimmunoassay data of insulin was found to be satisfactory.

  10. pDeep: Predicting MS/MS Spectra of Peptides with Deep Learning.

    Science.gov (United States)

    Zhou, Xie-Xuan; Zeng, Wen-Feng; Chi, Hao; Luo, Chunjie; Liu, Chao; Zhan, Jianfeng; He, Si-Min; Zhang, Zhifei

    2017-12-05

    In tandem mass spectrometry (MS/MS)-based proteomics, search engines rely on comparison between an experimental MS/MS spectrum and the theoretical spectra of the candidate peptides. Hence, accurate prediction of the theoretical spectra of peptides appears to be particularly important. Here, we present pDeep, a deep neural network-based model for the spectrum prediction of peptides. Using the bidirectional long short-term memory (BiLSTM), pDeep can predict higher-energy collisional dissociation, electron-transfer dissociation, and electron-transfer and higher-energy collision dissociation MS/MS spectra of peptides with >0.9 median Pearson correlation coefficients. Further, we showed that intermediate layer of the neural network could reveal physicochemical properties of amino acids, for example the similarities of fragmentation behaviors between amino acids. We also showed the potential of pDeep to distinguish extremely similar peptides (peptides that contain isobaric amino acids, for example, GG = N, AG = Q, or even I = L), which were very difficult to distinguish using traditional search engines.

  11. IgE and IgG cross-reactivity among Lol p I and Lol p II/III. Identification of the C-termini of Lol p I, II, and III as cross-reactive structures.

    Science.gov (United States)

    van Ree, R; van Leeuwen, W A; van den Berg, M; Weller, H H; Aalberse, R C

    1994-04-01

    In this study, the homologous C-termini of Lol p I, Lol p II, and Lol p III were shown to contain cross-reactive B-cell epitopes. This was demonstrated by inhibition studies with purified Lol p I, II, and III and synthetic peptides of their C-termini. It was ruled out that the observed cross-reactivity was caused by cross-contamination of the purified allergens. Both human IgE and IgG bound to the C-terminus of Lol p I. These antibodies were cross-reactive with Lol p II and, more specifically, with its C-terminus. Within a small panel of allergic patients, no cross-reactivity with Lol p III was found. A hyperimmune polyclonal rabbit antiserum against Lol p I also recognized the Lol p I C-terminus. As for human antibodies, cross-reactivity with Lol p II and its C-terminus was demonstrated. Cross-reactivity with Lol p III was demonstrated with C-terminal peptides, but not with native Lol p III. A polyclonal rabbit antiserum against Lol p II bound to the C-terminal peptides of both Lol p II and III. This binding was inhibited with Lol p I, confirming that cross-reactive structures exist not only on the C-termini of Lol p II and Lol p I, but also of Lol p III and Lol p I. The existence of cross-reactivity between Lol p I and Lol p II and III possibly contributes to the frequently observed cosensitization for these allergens in grass-pollen-allergic patients.

  12. Antiviral activity of a Bacillus sp: P34 peptide against pathogenic viruses of domestic animals

    Directory of Open Access Journals (Sweden)

    Débora Scopel e Silva

    2014-09-01

    Full Text Available P34 is an antimicrobial peptide produced by a Bacillus sp. strain isolated from the intestinal contents of a fish in the Brazilian Amazon basin with reported antibacterial activity. The aim of this work was to evaluate the peptide P34 for its in vitro antiviral properties against canine adenovirus type 2 (CAV-2, canine coronavirus (CCoV, canine distemper virus (CDV, canine parvovirus type 2 (CPV-2, equine arteritis virus (EAV, equine influenza virus (EIV, feline calicivirus (FCV and feline herpesvirus type 1 (FHV-1. The results showed that the peptide P34 exhibited antiviral activity against EAV and FHV-1. The peptide P34 inhibited the replication of EAV by 99.9% and FHV-1 by 94.4%. Virucidal activity was detected only against EAV. When P34 and EAV were incubated for 6 h at 37 °C the viral titer reduced from 10(4.5 TCID50 to 10(2.75 TCID50, showing a percent of inhibition of 98.6%. In conclusion, our results demonstrated that P34 inhibited EAV and FHV-1 replication in infected cell cultures and it showed virucidal activity against EAV. Since there is documented resistance to the current drugs used against herpesviruses and there is no treatment for equine viral arteritis, it is advisable to search for new antiviral compounds to overcome these infections.

  13. Blood Contamination in Saliva: Impact on the Measurement of Salivary Oxidative Stress Markers

    Directory of Open Access Journals (Sweden)

    Natália Kamodyová

    2015-01-01

    Full Text Available Salivary oxidative stress markers represent a promising tool for monitoring of oral diseases. Saliva can often be contaminated by blood, especially in patients with periodontitis. The aim of our study was to examine the impact of blood contamination on the measurement of salivary oxidative stress markers. Saliva samples were collected from 10 healthy volunteers and were artificially contaminated with blood (final concentration 0.001–10%. Next, saliva was collected from 12 gingivitis and 10 control patients before and after dental hygiene treatment. Markers of oxidative stress were measured in all collected saliva samples. Advanced oxidation protein products (AOPP, advanced glycation end products (AGEs, and antioxidant status were changed in 1% blood-contaminated saliva. Salivary AOPP were increased in control and patients after dental treatment (by 45.7% and 34.1%, p<0.01. Salivary AGEs were decreased in patients after microinjury (by 69.3%, p<0.001. Salivary antioxidant status markers were decreased in both control and patients after dental treatment (p<0.05 and p<0.01. One % blood contamination biased concentrations of salivary oxidative stress markers. Saliva samples with 1% blood contamination are visibly discolored and can be excluded from analyses without any specific biochemic detection of blood constituents. Salivary markers of oxidative stress were significantly altered in blood-contaminated saliva in control and patients with gingivitis after dental hygiene treatment.

  14. Cytosolically expressed PrP GPI-signal peptide interacts with mitochondria.

    Science.gov (United States)

    Guizzunti, Gianni; Zurzolo, Chiara

    2015-01-01

    We previously reported that PrP GPI-anchor signal peptide (GPI-SP) is specifically degraded by the proteasome. Additionally, we showed that the point mutation P238S, responsible for a genetic form of prion diseases, while not affecting the GPI-anchoring process, results in the accumulation of PrP GPI-SP, suggesting the possibility that PrP GPI-anchor signal peptide could play a role in neurodegenerative prion diseases. We now show that PrP GPI-SP, when expressed as a cytosolic peptide, is able to localize to the mitochondria and to induce mitochondrial fragmentation and vacuolarization, followed by loss in mitochondrial membrane potential, ultimately resulting in apoptosis. Our results identify the GPI-SP of PrP as a novel candidate responsible for the impairment in mitochondrial function involved in the synaptic pathology observed in prion diseases, establishing a link between PrP GPI-SP accumulation and neuronal death.

  15. Potent anti-tumor effect generated by a novel human papillomavirus (HPV antagonist peptide reactivating the pRb/E2F pathway.

    Directory of Open Access Journals (Sweden)

    Cai-ping Guo

    Full Text Available Human papillomavirus type 16 (HPV16 E7 is a viral oncoprotein believed to play a major role in cervical cancer. In this study, an antagonist peptide against HPV16E7 protein was first identified from screening the c7c phage display peptide library. The binding specificity and affinity of the selected peptide to HPV16E7 were tested by competitive enzyme-linked immunosorbent assay (ELISA. The antagonist peptide showed obvious anti-tumor efficacy both in cell lines and animal tumor models. Significant cell proliferation inhibition with high specificity was noted when HPV16-positive cells were treated with the peptide. This anti-tumor efficacy was resulted from overriding the activities of HPV16E7 and reactivating the pRb/E2F pathway, as shown by a series of experiments. Flow cytometry analysis revealed that the selected peptide induced G1 arrest in a dose-dependent manner. Competitive ELISA, pull down, and Co-IP experiments indicated that the selected peptide disrupted the interaction between HPV16E7 and pRb proteins both in vitro and in vivo. Luciferase reporter assay verified that transcription activities of E2F were suppressed by the peptide through restoration of pRb. RT-PCR and Western blot revealed that it reduced cyclins A, D1, and E1 expression, and led to HPV16E7 protein degradation, but pRb protein stabilization. The current study suggests that this specific peptide may serve as a potential therapeutic agent for HPV16-positive cervical cancer.

  16. Whole Saliva has a Dual Role on the Adherence of Candida albicans to Polymethylmetacrylate.

    Science.gov (United States)

    Elguezabal, N; Maza, J L; Dorronsoro, S; Pontón, J

    2008-01-01

    Adhesion of Candida albicans to acrylic of dental prostheses or to salivary macromolecules adsorbed on their surface is believed to be a critical event in the development of denture stomatitis. In previous studies our group has shown that adhesion of C. albicans germ tubes to polystyrene is decreased by saliva whereas C. albicans yeast cells adhesion to the same material is enhanced. The results presented in this study confirm this dual role played by whole saliva, since it decreased the adhesion of germ tubes but increased the adhesion of yeast cells to polymethylmetacrylate (PMMA). These effects mediated by whole saliva do not seem to be related to an inhibition of the germination of C. albicans, since similar levels of filamentation were observed in presence and absence of saliva. These results may give new insights into the conflicting role of saliva in the adhesion of C. albicans to acrylic resins of dental prostheses.

  17. Factors determining the passage of drugs from blood into saliva.

    OpenAIRE

    Stephen, K W; McCrossan, J; Mackenzie, D; Macfarlane, C B; Speirs, C F

    1980-01-01

    1. Following single oral dosing of ampicillin, cephalexin, tetracycline, erythromycin estolate, clindamycin and rifampicin to six normal volunteers, antibacterial activity was measured at 1, 3 and 6 h in serum, gingival fluid and minor gland saliva from all subjects and in parotid and submandiabular saliva from three. 2. pH values of all gingival fluid and saliva specimens were noted. 3. Partition coefficients between n-octanol and water were measured for erythromycin, clindamycin and rifampi...

  18. A 13C{31P} REDOR NMR Investigation of the Role of Glutamic Acid Residues in Statherin-Hydroxyapatite Recognition

    Science.gov (United States)

    Ndao, Moise; Ash, Jason T.; Breen, Nicholas F.; Goobes, Gil; Stayton, Patrick S.; Drobny, Gary P.

    2011-01-01

    The side chain carboxyl groups of acidic proteins found in the extra-cellular matrix (ECM) of mineralized tissues play a key role in promoting or inhibiting the growth of minerals such as hydroxyapatite (HAP), the principal mineral component of bone and teeth. Among the acidic proteins found in the saliva is statherin, a 43-residue tyrosine-rich peptide that is a potent lubricant in the salivary pellicle and an inhibitor of both HAP crystal nucleation and growth. Three acidic amino acids – D1, E4, and E5 – are located in the N-terminal 15 amino acid segment, with a fourth amino acid, E26, located outside the N-terminus. We have utilized 13C{31P} REDOR NMR to analyze the role played by acidic amino acids in the binding mechanism of statherin to the HAP surface by measuring the distance between the δ-carboxyl 13C spins of the three glutamic acid side chains of statherin (residues E4, E5, E26) and 31P spins of the phosphate groups at the HAP surface. 13C{31P} REDOR studies of glutamic-5-13C acid incorporated at positions E4 and E26 indicate a 13C–31P distance of more than 6.5 Å between the side chain carboxyl 13C spin of E4 and the closest 31P in the HAP surface. In contrast, the carboxyl 13C spin at E5 has a much shorter 13C–31P internuclear distance of 4.25±0.09 Å, indicating that the carboxyl group of this side chain interacts directly with the surface. 13C T1ρ and slow-spinning MAS studies indicate that the motions of the side chains of E4 and E5 are more restricted than that of E26. Together, these results provide further insight into the molecular interactions of statherin with HAP surfaces. PMID:19678690

  19. Microstructure and nanomechanical properties of enamel remineralized with asparagine-serine-serine peptide

    Energy Technology Data Exchange (ETDEWEB)

    Chung, Hsiu-Ying, E-mail: hychung@mail.fcu.edu.tw; Li, Cheng Che

    2013-03-01

    A highly biocompatible peptide, triplet repeats of asparagine-serine-serine (3NSS) was designed to regulate mineral deposition from aqueous ions in saliva for the reconstruction of enamel lesions. Healthy human enamel was sectioned and acid demineralized to create lesions, then exposed to the 3NSS peptide solution, and finally immersed in artificial saliva for 24 h. The surface morphology and roughness were examined using scanning electron microscopy (SEM) and atomic force microscopy (AFM), respectively. X-ray diffraction (XRD) was used to identify the phases and crystallinity of the deposited minerals observed on the enamel surface. Attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) was used to quantitatively analyze the mineral variation by calculating the relative integrated-area of characteristic bands. Nanohardness and elastic modulus measured by nanoindentation at various treatment stages were utilized to evaluate the degree of recovery. Biomimetic effects were accessed according to the degree of nanohardness recovery and the amount of hydroxyapatite deposition. The charged segments in the 3NSS peptide greatly attracted aqueous ions from artificial saliva to form hydroxyapatite crystals to fill enamel caries, in particular the interrod areas, resulting in a slight reduction in overall surface roughness. Additionally, the deposited hydroxyapatites were of a small crystalline size in the presence of the 3NSS peptide, which effectively restrained the plastic deformations and thus resulted in greater improvements in nanohardness and elastic modulus. The degree of nanohardness recovery was 5 times greater for remineralized enamel samples treated with the 3NSS peptide compared to samples without peptide treatment. - Highlights: Black-Right-Pointing-Pointer The degree of nanohardness recovery of enamel was 4 times greater with the aid of 3NSS peptide. Black-Right-Pointing-Pointer 3NSS peptide promoted the formation of hydroxyapatites with

  20. Different host complement systems and their interactions with saliva from Lutzomyia longipalpis (Diptera, Psychodidae) and Leishmania infantum promastigotes.

    Science.gov (United States)

    Mendes-Sousa, Antonio Ferreira; Nascimento, Alexandre Alves Sousa; Queiroz, Daniel Costa; Vale, Vladimir Fazito; Fujiwara, Ricardo Toshio; Araújo, Ricardo Nascimento; Pereira, Marcos Horácio; Gontijo, Nelder Figueiredo

    2013-01-01

    Lutzomyia longipalpis is the vector of Leishmania infantum in the New World, and its saliva inhibits classical and alternative human complement system pathways. This inhibition is important in protecting the insect´s midgut from damage by the complement. L. longipalpis is a promiscuous blood feeder and must be protected against its host's complement. The objective of this study was to investigate the action of salivary complement inhibitors on the sera of different host species, such as dogs, guinea pigs, rats and chickens, at a pH of 7.4 (normal blood pH) and 8.15 (the midgut pH immediately after a blood meal). We also investigated the role of the chicken complement system in Leishmania clearance in the presence and absence of vector saliva. The saliva was capable of inhibiting classical pathways in dogs, guinea pigs and rats at both pHs. The alternative pathway was not inhibited except in dogs at a pH of 8.15. The chicken classical pathway was inhibited only by high concentrations of saliva and it was better inhibited by the midgut contents of sand flies. Neither the saliva nor the midgut contents had any effect on the avian alternative pathway. Fowl sera killed L. infantum promastigotes, even at a low concentration (2%), and the addition of L. longipalpis saliva did not protect the parasites. The high body temperature of chickens (40°C) had no effect on Leishmania viability during our assays. Salivary inhibitors act in a species-specific manner. It is important to determine their effects in the natural hosts of Leishmania infantum because they act on canid and rodent complements but not on chickens (which do not harbour the parasite). Moreover, we concluded that the avian complement system is the probable mechanism through which chickens eliminate Leishmania and that their high body temperature does not influence this parasite.

  1. Short-term glucagon stimulation test of C-peptide effect on glucose utilization in patients with type 1 diabetes mellitus.

    Science.gov (United States)

    Mojto, Viliam; Rausova, Zuzana; Chrenova, Jana; Dedik, Ladislav

    2015-12-01

    This work aimed to evaluate the use of a four-point glucagon stimulation test of C-peptide effect on glucose utilization in type 1 diabetic patients using a new mathematical model. A group of 32 type 1 diabetic patients and a group of 10 healthy control subjects underwent a four-point glucagon stimulation test with blood sampling at 0, 6, 15 and 30 min after 1 mg glucagon bolus intravenous administration. Pharmacokinetic and pharmacokinetic/pharmacodynamic models of C-peptide effect on glucose utilization versus area under curve (AUC) were used. A two-sample t test and ANOVA with Bonferroni correction were used to test the significance of differences between parameters. A significant difference between control and patient groups regarding the coefficient of whole-body glucose utilization and AUC C-peptide/AUC glucose ratio (p ≪ 0.001 and p = 0.002, respectively) was observed. The high correlation (r = 0.97) between modeled coefficient of whole-body glucose utilization and numerically calculated AUC C-peptide/AUC glucose ratio related to entire cohort indicated the stability of used method. The short-term four-point glucagon stimulation test allows the numerically calculated AUC C-peptide/AUC glucose ratio and/or the coefficient of whole-body glucose utilization calculated from model to be used to diagnostically identify type 1 diabetic patients.

  2. Immediate Repair Bond Strength of Fiber-reinforced Composite after Saliva or Water Contamination.

    Science.gov (United States)

    Bijelic-Donova, Jasmina; Flett, Andrew; Lassila, Lippo V J; Vallittu, Pekka K

    2018-05-31

    This in vitro study aimed to evaluate the shear bond strength (SBS) of particulate filler composite (PFC) to saliva- or water-contaminated fiber-reinforced composite (FRC). One type of FRC substrate with semi-interpenetrating polymer matrix (semi-IPN) (everStick C&B) was used in this investigation. A microhybrid PFC (Filtek Z250) substrate served as control. Freshly cured PFC and FRC substrates were first subjected to different contamination and surface cleaning treatments, then the microhybrid PFC restorative material (Filtek Z250) was built up on the substrates in 2-mm increments and light cured. Uncontaminated and saliva- or water-contaminated substrate surfaces were either left untreated or were cleaned via phosphoric acid etching or water spray accompanied with or without adhesive composite application prior applying the adherent PFC material. SBS was evaluated after thermocycling the specimens (6000 cycles, 5°C and 55°C). Three-way ANOVA showed that both the surface contamination and the surface treatment signficantly affected the bond strength (p contamination reduced the SBS more than did the water contamination. SBS loss after saliva contamination was 73.7% and 31.3% for PFC and FRC, respectively. After water contamination, SBS loss was 17.2% and 13.3% for PFC and FRC, respectively. The type of surface treatment was significant for PFC (p contamination of freshly cured PFC or semi-IPN FRC, surfaces should be re-prepared via phosphoric acid etching, water cleaning, drying, and application of adhesive composite in order to recover optimal bond strength.

  3. Clinical aspects of Candida species carriage in saliva of xerotomic subjects.

    Science.gov (United States)

    Torres, S R; Peixoto, C B; Caldas, D M; Silva, E B; Magalhães, F A C; Uzeda, M; Nucci, M

    2003-10-01

    In order to investigate the clinical factors that might influence the diversity and the degree of Candida species carriage in saliva, we conducted a cross-sectional study with 133 patients with complaints of xerostomia. Anamnesis, oral examination and collection of chewing-stimulated whole saliva were performed. The samples of saliva were kept refrigerated until they were plated onto CHROMagar Candida; cfu were counted and Candida species were identified by standard methods. There was a high prevalence of mixed Candida colonization. No relationship was found between total Candida cfu counts and variables like gender, age, place of origin, underlying diseases, exposure to medications (except antibiotics), daily habits and salivary flow rates. Oral candidiasis, antibiotic exposure and dental prosthesis wearing were associated with relatively high Candida counts in saliva. Low salivary flow rates predisposed to intense colonization by C. albicans and C. parapsilosis.

  4. C-Peptide Is Independently Associated with an Increased Risk of Coronary Artery Disease in T2DM Subjects: A Cross-Sectional Study.

    Directory of Open Access Journals (Sweden)

    Lingshu Wang

    Full Text Available C-peptide has been reported to be a marker of subclinical atherosclerosis in type 2 diabetes mellitus (T2DM patients, whereas its role in coronary artery disease (CAD has not been clarified, especially in diabetics with differing body mass indices (BMIs.This cross-sectional study included 501 patients with T2DM. First, all subjects were divided into the following two groups: CAD and non-CAD. Then, binary logistic regression was used to determine the risk factors for CAD for all patients. To clarify the role of obesity, we re-divided all subjects into two additional groups (obese and non-obese based on BMI. Finally, binary logistic regression was used to determine the risk factors for CAD for each weight group.The patients with CAD showed a higher BMI and fasting C-peptide level in addition to an increased prevalence of traditional risk factors for CAD, such as hypertension, insulin resistance, higher cholesterol, cysteine-C (Cys-C and lower estimated glomerular filtration rate (eGFR. Logistic regression analysis showed that fasting C-peptide (OR=1.513, p=0.005, insulin treatment (OR=1.832, p=0.027 hypertension (OR=1.987, p=0.016 and hyperlipidemia (OR=4.159, p<0.001 significantly increased the risk of clinical CAD in the T2DM patients independent of age, gender, diabetes duration, smoking and alcohol statuses, fasting insulin and glucose, hypoglycemic episodes, UA and eGFR. Additionally, in both of the obese (OR=1.488, p=0.049 and non-obese (OR=1.686, p=0.037 DM groups, C-peptide was associated with an increased risk of CAD after multiple adjustments.C-peptide is associated with an increased CAD risk in T2DM patients, no matter whether they are obese or not.

  5. Study of pH (low) insertion peptides (pHLIPs) interaction with lipid bilayer of membrane

    Science.gov (United States)

    Weerakkody, Dhammika

    The pH-dependent interactions of pHLIPsRTM (pH (Low) Insertion Peptides) with lipid bilayer of membrane provides an opportunity to study and address fundamental questions of protein folding/insertion into membrane and unfolding/exit, as well as develop novel approach to target acidic diseased tissue such as cancer, ischemic myocardium, infection and others. The main goal of the work presented here is to answer the following questions: - What is the molecular mechanism of spontaneous insertion and folding of a peptide in a lipid bilayer of membrane; - What is the molecular mechanism of unfolding and exit of a peptide from a lipid bilayer of membrane; - How polar cargo attached to a peptide's inserting end might affect the process of insertion into a lipid bilayer of membrane; How sequence variation will affect a peptide's interactions with a lipid bilayer of membrane (partitioning into bilayer at neutral and low pH; apparent pK of insertion) with the main goal to identify the best pHLIP variants for imaging and therapy of pathological states such as cancer and others. It has been demonstrated that pHLIP insertion into a membrane is associated with the protonation of Asp/Glu residues, which leads to an increase of hydrophobicity that triggers the folding and insertion of the peptide across a lipid bilayer. The insertion of the pHLIP is unidirectional and it is accompanied by the release of energy. Therefore, the energy of membrane associated-folding can be used to favor the movement of cell-impermeable polar cargo molecules across the hydrophobic membrane bilayer when they are attached to the inserting end of pHLIP. Both pH-targeting behavior and molecular translocation have been demonstrated in cultured cells and in vivo. Thus, there is an opportunity to develop a novel concept in drug delivery, which is based on the use of a monomeric, pH-sensitive peptide molecular transporter, to deliver agents that are significantly more polar than conventional drugs

  6. Saliva secretion difference before and after rinsing with baking soda on menopause women

    Directory of Open Access Journals (Sweden)

    Dewi Anggraeni

    2007-03-01

    Full Text Available Menopause women can experience a decrease in saliva secretion (decrease. To understand the clear picture about saliva secretion, the volume, flow rate, pH and viscosity were then measured. The aim of this research was to obtain a picture about the difference of saliva secretion before and after rinsing with baking soda on menopause women. The type of the research used was a laboratory quasi-experiment with comparative descriptive form. The technique used in this research is the survey method, and samples were taken using the multistage cluster random sampling method, and t-student statistical analysis. This research was conducted with the saliva collected with spitting method on 45 menopause women. The results show that the average volume, flow rate, pH and viscosity before rinsing with baking soda was 1.79 ml, 0.18 ml/minute, 7.40 and 0.81 mm2/second. The average volume, flow rate, pH and viscosity after rinsing with baking soda were 2.66 ml; 0.27 ml/minute; 8.67 and 0.78 mm2/second. Statistical analysis t-student on α = 0.05 shows volume changes, flow rate, pH and saliva viscosity before and after rinsing with baking soda was 0.873; 0.086; 1.273 and 0.037 respectively. The conclusion shows a significant difference between saliva secretion before and after rinsing with baking soda, and saliva secretion after rinsing with baking soda on menopause women.

  7. Different Dose-Dependent Modes of Action of C-Type Natriuretic Peptide on Pseudomonas aeruginosa Biofilm Formation

    Directory of Open Access Journals (Sweden)

    Florie Desriac

    2018-04-01

    Full Text Available We have previously shown that the C-type Natriuretic Peptide (CNP, a peptide produced by lungs, is able to impact Pseudomonas aeruginosa physiology. In the present work, the effect of CNP at different concentrations on P. aeruginosa biofilm formation was studied and the mechanisms of action of this human hormone on P. aeruginosa were deciphered. CNP was shown to inhibit dynamic biofilm formation in a dose-dependent manner without affecting the bacterial growth at any tested concentrations. The most effective concentrations were 1 and 0.1 µM. At 0.1 µM, the biofilm formation inhibition was fully dependent on the CNP sensor protein AmiC, whereas it was only partially AmiC-dependent at 1 µM, revealing the existence of a second AmiC-independent mode of action of CNP on P. aeruginosa. At 1 µM, CNP reduced both P. aeruginosa adhesion on glass and di-rhamnolipid production and also increased the bacterial membrane fluidity. The various effects of CNP at 1 µM and 0.1 µM on P. aeruginosa shown here should have major consequences to design drugs for biofilm treatment or prevention.

  8. Synthesis and screening of peptide libraries with free C-termini.

    Science.gov (United States)

    Wang, Yen-Chih; Distefano, Mark D

    2014-01-01

    Peptide libraries are useful tools to investigate the relationship between structure and function of proteins. The creation of peptide libraries with free C-termini presents unique synthetic challenges. In this review, methods for creating peptide libraries using either solid-phase peptide synthesis or phage display are described. Methods for screening such libraries and their application in studying several important biological problems are also reported.

  9. Candida in saliva of Brazilian hemophilic patients Candida na saliva de pacientes hemofílicos brasileiros

    Directory of Open Access Journals (Sweden)

    Claudio Maranhão Pereira

    2004-12-01

    Full Text Available Hemophilia is a common hereditary hemorrhagic disorder, however little is known about the oral microflora of hemophilic patients. The aim of this study was to quantify the Candida and identify its species in non-stimulated saliva of hemophilic patients, and consider its relationship with clinical factors influencing Candida carriage. This study comprised evaluation of 86 hemophilic patients of the Hematology Center/UNICAMP and 43 healthy subjects as controls. All patients were submitted to anamnesis, intraoral examination and unstimulated saliva collection. Candida counts and species identification were performed in salivary samples. Candida was present in 64% of the hemophilic patients and in 44% of the healthy controls. C. albicans represented 65% and 68% of the isolated species, in hemophiliacs and control group respectively, and C. tropicalis was the second most common species in both groups. These results indicate that hemophilic patients carry Candida more frequently and in higher counts than healthy controls, independently of oral clinical parameter considered, as viral infections, complete dentures, transfusions of hemoderivatives, and salivary flow.Hemofilia é uma alteração hemorrágica hereditária comum, entretanto pouco se sabe a respeito da microbiota oral destes indivíduos. O objetivo deste estudo foi quantificar a presença de Candida e identificar as suas espécies na saliva de hemofílicos, correlacionando os resultados com fatores clínicos que possam influenciar a presença deste fungo. Foram avaliados 86 hemofílicos do Hemocentro/UNICAMP e 43 indivíduos saudáveis. Todos os pacientes foram submetidos a anamnese, exame clínico intra-oral e coleta de saliva de forma não estimulada. A quantificação e identificação das espécies de Candida foram realizadas nas amostras de saliva. Candida estava presente em 64% dos hemofílicos e em 44% dos indivíduos saudáveis. C. albicans representou 65% e 68% das esp

  10. [The physicochemical and microbiological characteristics of saliva during and after pregnancy].

    Science.gov (United States)

    Martínez-Pabón, María C; Martínez Delgado, Cecilia M; López-Palacio, Ana M; Patiño-Gómez, Lina M; Arango-Pérez, Eduin A

    2014-01-01

    Identify the changes in some physiological and microbiological parameters in the saliva from a group of women during and after their pregnancies. Stimulated whole saliva was collected from a cohort of 35 women during their pregnancy and afterwards to determine each sample's physicochemical (secretion rate, pH and buffer capacity) and microbiological characteristics (acidogenic bacteria count). The pH and buffer capacity of saliva during pregnancy were lower than after pregnancy. There were no statistically significant changes regarding S. mutans and Lactobacillus spp. count, but a tendency towards increased values during pregnancy was noted. Changes occurring in the saliva of pregnant women can lead to an increase of risk of suffering disease affecting one's oral health, such as caries, gingivitis and periodontal disease; this could be prevented by appropriate diagnosis and dental follow-up, including education regarding pregnant women's oral health.

  11. Investigation of trefoil factor expression in saliva and oral mucosal tissues of patients with oral squamous cell carcinoma

    DEFF Research Database (Denmark)

    Chaiyarit, Ponlatham; Utrawichian, Akasith; Leelayuwat, Chanvit

    2012-01-01

    Objectives The aims of our study were to determine levels of trefoil factor (TFF) peptides in saliva and oral mucosal tissues from patients with oral squamous cell carcinoma (OSCC), and to evaluate whether individual members of TFFs (TFF1, TFF2, and TFF3) might act as biomarkers of disease....... Materials and methods Saliva samples were from 23 healthy subjects and 23 OSCC patients. Tissue samples were collected from 32 normal oral mucosa (NOM) and 32 OSCC biopsy specimens. ELISA and immunohistochemical methods were used to evaluate the expression of TFF1, TFF2, and TFF3 in saliva and oral mucosal...... tissues, respectively. Results Expression of TFF2 and TFF3 in oral mucosal tissues of OSCC patients was strongly downregulated when compared to healthy subjects (p 

  12. A high-throughput mass spectrometry assay to simultaneously measure intact insulin and C-peptide.

    Science.gov (United States)

    Taylor, Steven W; Clarke, Nigel J; Chen, Zhaohui; McPhaul, Michael J

    2016-04-01

    Measurements of fasting levels of insulin and C-peptide are useful in documenting insulin resistance and may help predict development of diabetes mellitus. However, the specific insulin and C-peptide levels associated with specific degrees of insulin resistance have not been defined, owing to marked variability among immunoassays and lack of standardization. Herein, we describe a multiplexed liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for intact insulin and C-peptide. Insulin and C-peptide were enriched from patient sera using monoclonal antibodies immobilized on magnetic beads and processed on a robotic liquid handler. Eluted peptides were analyzed by LC-MS/MS. Bovine insulin and a stable isotopically-labeled (13C/15N) C-peptide were utilized as internal standards. The assay had an analytical measurement range of 3 to 320 μIU/ml (18 to 1920 pmol/l) for insulin and 0.11 to 27.2 ng/ml (36 to 9006 pmol/l) for C-peptide. Intra- and inter-day assay variation was less than 11% for both peptides. Of the 5 insulin analogs commonly prescribed to treat diabetes, only the recombinant drug insulin lispro caused significant interference for the determination of endogenous insulin. There were no observed interferences for C-peptide. We developed and validated a high-throughput, quantitative, multiplexed LC-MS/MS assay for intact insulin and C-peptide. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Correlation between factors associated with the removable partial dentures use and Candida spp. in saliva.

    Science.gov (United States)

    Gusmão, João Milton Rocha; Ferreira dos Santos, Silvana Soleo; Neisser, Maximiliano Piero; Jorge, Antônio Olavo Cardoso; Faria, Ms Ivan

    2011-12-01

    To correlate the presence and number of Candida spp. in the saliva of wearers of removable partial dentures retained with precision attachments with the proportion of metal/acrylic resin present in the dentures. Saliva samples from 40 removable partial denture wearers (test) and one paired sample of individuals, non-wearers of any type of removable denture (control) were collected, seeded, and the colony forming units of Candida counted and identified. The metal/acrylic resin proportion of each denture was quantified, using silicone plates pressed over each denture. Candida spp. was found in the saliva of 80% of the individuals in the test group and 65% of the control, with C. albicans being the most prevalent species. The test group presented with the highest number of colony forming units of Candida per ml of saliva, and there was weak correlation between this number and the metal and resin area of the denture (Pearson's coefficient of correlation). Greater prevalence and a higher number of colony forming units of Candida per ml of saliva occurred in removable partial denture wearers (p = 0.04) with a weak positive correlation between the metal and resin area and the number of colony forming units of Candida per ml of saliva. However, this correlation was more significant for the area of resin. © 2010 The Gerodontology Society and John Wiley & Sons A/S.

  14. Correlation of the secretion of insulin and C-peptide in cancer patients

    Energy Technology Data Exchange (ETDEWEB)

    Modnikov, O P; Lomtev, N G [Kirgizskij Nauchno-Issledovatel' skij Inst. Onkologii i Radiologii, Frunze (USSR)

    1983-08-01

    Insulin and C-peptide levels were studied with a radioimmunoassay in the peripheral blood serum of 44 patients with gastric and cervical cancer and 22 healthy persons. Hyperfunction of the pancreatic insular apparatus was shown in cancer patients which was expressed in a statistically significant increase in the C-peptide level. In gastric cancer patients hyperfunction of the insular apparatus was accompanied by hypoinsulinemia, and in cervical cancer patients by hormoinsulinemia. An analysis has shown that the ratio insulin/C-peptide in gastric and cervical cancer patients was about the same and significantly lower than the control. A conclusion has been made that in spite of difference in the initial insulin concentration, the same phenomenon - acceleration of the metabolic clearance of insulin - occurs in patients with cancer of the above sites. The C-peptide level decreased, the ratio insulin/C-peptide increased, i.e. hyperfunction of the insular apparatus disappeared and the metabolic clearance of insulin slowed down.

  15. Correlation of the sectetion of insulin and C-peptide in cancer patients

    International Nuclear Information System (INIS)

    Modnikov, O.P.; Lomtev, N.G.

    1983-01-01

    Insulin and C-peptide levels were studied with a radioimmunoassay in the peripheral blood serum of 44 patients with gastric and cervical cancer and 22 healthy persons. Hyperfunction of the pancreatic insular apparatus was shown in cancer patients which was expressed in a statistically significant increase in the C-peptide level. In gastric cancer patients hyperfunction of the insular apparatus was accompanied by hypoinsulinemia, and in cervical cancer patients by hormoinsulinemia. An analysis has shown that the ratio insulin/C-peptide in gastric and cervical cancer patients was about the same and significantly lower than the control. A conclusion has been made that in spite of difference in the initial insulin concentration, the same phenomenon - acceleration of the metabolic clearance of insulin - occurs in patients with cancer of the above sites. The C-peptide level decreased, the ratio insulin/C-peptide increased, i.e. hyperfunction of the insular apparatus disappeared and the metabolic clearance of insulin slowed down

  16. Trace element measurement in Saliva by NAA and PIXE techniques

    Energy Technology Data Exchange (ETDEWEB)

    Hamidian, M.R.; Vahid Golpayegani, M.; Shojai, S. (Shahid Beheshti Medical Science Univ., Shemiran, Tehran (Iran, Islamic Republic of))

    1993-01-01

    The activity of salivary glands and the chemical and physical properties of saliva, especially in some illnesses in which the activity of salivary glands and the chemical and physical properties alter, sometimes have severe effects on sedimentation and tooth decay. Long-standing investigations have shown the relationship between salivary gland activity and saliva composition in dental carries. Many modern techniques have been employed to measure important elements in saliva. The major elements in saliva include sodium, potassium, calcium, magnesium, chlorine, phosphorus, iodine, and fluorine. It should be pointed out that the amount of minerals changes when the diet changes. The major constituent of saliva is water with a density of 1.007 g/cm[sup 3] in which 0.6% is solid, 0.3% organic material and 0.3% inorganic material. In addition to other effects, the acidity (pH) of saliva has a strong effect on tooth sedimentation. Type of work, degree of stress, and mental condition affect salivary gland activity. When the acidity of salivary fluid in the mouth and consequently over the teeth drops, sedimentation increases. In this paper, the results of trace element measurement in saliva are presented.

  17. Interactions between resin monomers and commercial composite resins with human saliva derived esterases.

    Science.gov (United States)

    Jaffer, F; Finer, Y; Santerre, J P

    2002-04-01

    Cholesterol esterase (CE) and pseudocholinesterase (PCE) have been reported to degrade commercial and model composite resins containing bisphenylglycidyl dimethacrylate (BisGMA), triethylene glycol dimethacrylate (TEGDMA) or the latter in combination with urethane modified BisGMA monomer systems. In addition, human saliva has been shown to contain esterase like activities similar to CE and PCE. Hence, it was the aim of the current study to determine to what extent human saliva could degrade two common commercial composite resins (Z250 from 3M Inc. and Spectrum TPH from L.D. Caulk) which contain the above monomer systems. Saliva samples from different volunteers were collected, processed, pooled, and freeze-dried. TEGDMA and BisGMA monomers were incubated with human saliva derived esterase activity (HSDEA) and their respective hydrolysis was monitored using high performance liquid chromatography (HPLC). Both monomers were completely hydrolyzed within 25 h by HSDEA. Photopolymerized composites were incubated with buffer or human saliva (pH 7.0 and 37 C) for 2, 8 and 16 days. The incubation solutions were analyzed using HPLC and mass spectrometry. Surface morphology characterization was carried out using scanning electron microscopy. Upon biodegradation, the Z250 composite yielded higher amounts of BisGMA and TEGDMA related products relative to the TPH composite. However, there were higher amounts of ethoxylated bis-phenol A released from the TPH material. In terms of total mass of products released, human saliva demonstrated a greater ability to degrade Z250. In summary, HSDEA has been shown to contain esterase activities that can readily catalyze the biodegradation of current commercial composite resins.

  18. Anti-inflammatory and antinociceptive activities of Rhipicephalus microplus saliva

    Directory of Open Access Journals (Sweden)

    D F Buccini

    2018-01-01

    Full Text Available Objective: To evaluate the antinociceptive and anti-inflammatory activities and the toxic effects of Rhipicephalus microplus saliva for elucidating the modulation mechanism between arthropod saliva and host. Methods: For saliva collection, engorged ticks were obtained from a controlled bovine infestation and collected by natural fall. The ticks were fixed and injected pilocarpine 0.2% for induction of salivation. Saliva was collected, lyophilized and stored at - 80 °C. Cytotoxic activity was assessed by the hemolysis method (25, 50, 100, 200 and 300 μ g/mL and MTT cell viability assay (2.5, 5, 10, 20 and 40 μ g/mL for 24, 48 and 72 h. Anti-inflammatory activity was evaluated using the method of neutrophil migration to the peritoneal cavity of mice at doses of 10, 15 and 20 mg/kg; antinociceptive activity was assessed using the acetic acid-induced writhing test, and formalin-induced paw-licking in mice at dose of 15 mg/kg. Results: Saliva did not cause erythrocytes hemolysis at any concentration tested, as well as did not decrease cell viability in the MTT assay. Saliva inhibited neutrophil migration by 87% and 73% at doses of 15 and 20 mg/kg, respectively. In the nociceptive tests, saliva presented analgesic activity of 69.96% in the abdominal writhing test, and of 84.41% in the formalin test. Conclusions: The study proves that Rhipicephalus microplus saliva has significant in vivo anti-inflammatory and antinociceptive activities. The data presented herein support the development of further studies to elucidate the active principles of Rhipicephalus microplus saliva and its mechanism of action and, in future, to develop novel anti-inflammatory and analgesic drugs.

  19. Saliva Preservative for Diagnostic Purposes

    Science.gov (United States)

    Pierson, Duane L.; Mehta, Satish K.

    2012-01-01

    Saliva is an important body fluid for diagnostic purposes. Glycoproteins, glucose, steroids, DNA, and other molecules of diagnostic value are found in saliva. It is easier to collect as compared to blood or urine. Unfortunately, saliva also contains large numbers of bacteria that can release enzymes, which can degrade proteins and nucleic acids. These degradative enzymes destroy or reduce saliva s diagnostic value. This innovation describes the formulation of a chemical preservative that prevents microbial growth and inactivates the degradative enzymes. This extends the time that saliva can be stored or transported without losing its diagnostic value. Multiple samples of saliva can be collected if needed without causing discomfort to the subject and it does not require any special facilities to handle after it is collected.

  20. Lipopolysaccharide interactions of C-terminal peptides from human thrombin.

    Science.gov (United States)

    Singh, Shalini; Kalle, Martina; Papareddy, Praveen; Schmidtchen, Artur; Malmsten, Martin

    2013-05-13

    Interactions with bacterial lipopolysaccharide (LPS), both in aqueous solution and in lipid membranes, were investigated for a series of amphiphilic peptides derived from the C-terminal region of human thrombin, using ellipsometry, dual polarization interferometry, fluorescence spectroscopy, circular dichroism (CD), dynamic light scattering, and z-potential measurements. The ability of these peptides to block endotoxic effects caused by LPS, monitored through NO production in macrophages, was compared to peptide binding to LPS and its endotoxic component lipid A, and to size, charge, and secondary structure of peptide/LPS complexes. While the antiendotoxic peptide GKY25 (GKYGFYTHVFRLKKWIQKVIDQFGE) displayed significant binding to both LPS and lipid A, so did two control peptides with either selected D-amino acid substitutions or with maintained composition but scrambled sequence, both displaying strongly attenuated antiendotoxic effects. Hence, the extent of LPS or lipid A binding is not the sole discriminant for the antiendotoxic effect of these peptides. In contrast, helix formation in peptide/LPS complexes correlates to the antiendotoxic effect of these peptides and is potentially linked to this functionality. Preferential binding to LPS over lipid membrane was furthermore demonstrated for these peptides and preferential binding to the lipid A moiety within LPS inferred.

  1. Development and evaluation of a saliva-based chair-side diagnostic for the detection of Porphyromonas gingivalis

    Directory of Open Access Journals (Sweden)

    Neil M. O'Brien-Simpson

    2015-09-01

    Full Text Available Porphyromonas gingivalis is a key pathogen in the polymicrobial biofilm that is associated with the oral disease chronic periodontitis. A number of studies have shown that in humans the level of P. gingivalis in the polymicrobial biofilm is positively correlated with disease progression. The aim of this study was to develop a P. gingivalis diagnostic that has high specificity and sensitivity for P. gingivalis using a range of laboratory and clinical isolates and then compare the efficacy of the diagnostic with RTPCR using samples from chronic periodontitis patients and age- and sex-matched healthy controls. Key parameters for the kit were to use saliva as the biological fluid as this is a most convenient medium for chair-side sampling and to give a positive reading for the reported threshold for detection of 5×105 P. gingivalis cells/mL that indicates disease progression. We initially screened a range of monoclonal antibodies for recognition of the P. gingivalis conserved virulence factor RgpA-Kgp complex and identified two mAbs that could be used in a capture and detection ELISA system. These mAbs were used to formulate and manufacture the GC P. gingivalis saliva diagnostic kit used in the study. To validate the saliva kit, saliva (P. gingivalis free was spiked with known concentrations of viable P. gingivalis whole cells of W50, 381, A7A1-28, and ATCC 33277; P. gingivalis clinical isolates; P. gingivalis vesicles; and the secreted form of the RgpA-Kgp complex. Laboratory findings indicated that the kit was able to detect all laboratory and clinical isolate strains of P. gingivalis at 5×104/mL to 5×105/mL. It was also able to detect the RgpA-Kgp complex and vesicles at 5×104 and 5×105 cell equivalent doses, respectively. Saliva and plaque were then collected from 50 subjects with moderate–severe chronic periodontitis and 50 age- and sex-matched subjects with healthy periodontium. Real-time PCR was utilised to analyse levels of P

  2. Total antioxidant property and pH change of dental plaque and saliva in 6-11-year-old children after consumption of flavored milk

    Directory of Open Access Journals (Sweden)

    Effat Khodadadi

    2013-09-01

    Full Text Available Introduction: The antioxidant properties of chocolate and other flavored additives besides the sugar added to milk raises the question about the acidogenecity of flavored milk. This study was conducted to measure the pH changes of dental plaque and saliva after the consumption of flavored milk and evaluate the antioxidant property of them. Methods: This study was performed on 42 samples of dental plaque and 42 samples of saliva in 6-11 year old school going children. Milk with flavors of strawberry, chocolate, banana, honey and slim milk were evaluated, all from the same manufacturer with a similar production date. At the beginning of the study on the first day, children were given thorough oral propHylaxis and they were instructed to avoid any method of oral hygiene for 48 hours to permit enough plaque deposition. On the third day the children were divided into 7 groups, 6 children in each group. The supra-gingival plaque was collected through the help of an excavator #3 which was pulled twice with the same force on the tooth surface. The saliva was collected using spitting technique. Each child swished 10 cc of milk for 1 minute in his/her mouth. Fresh plaque samples after 5, 10, 20 and 30 minutes and saliva samples immediately, after 5, 10, 20 and 30 minutes were collected. The pH of the samples were recorded by a pH testing apparatus (Basic 20+, Crisom. To evaluate the antioxidant property of studied milk, Frap test was performed. The collected readings were reported as mean±SD and analyzed by ANOVA repeated measures, Post hoc Tukey and Paired T-test. In this study, p≤0.05 was considered as significant. Results: After 30 minutes, honey milk caused the least drop 0.74±0.30 and banana milk caused the highest drop 1.38± 0.25 in plaque pH (p≤0.05. After 30 minutes, the pH of saliva showed no significant difference compared to the initial pH. Chocolate milk contained the highest (1000 micromol/liter and banana milk the lowest (706

  3. Salivary Parameters (Salivary Flow, pH and Buffering Capacity) in Stimulated Saliva of Mexican Elders 60 Years Old and Older.

    Science.gov (United States)

    Islas-Granillo, H; Borges-Yañez, S A; Medina-Solís, C E; Galan-Vidal, C A; Navarrete-Hernández, J J; Escoffié-Ramirez, M; Maupomé, G

    2014-12-01

    To compare a limited array of chewing-stimulated saliva features (salivary flow, pH and buffer capacity) in a sample of elderly Mexicans with clinical, sociodemographic and socio-economic variables. A cross-sectional study was carried out in 139 adults, 60 years old and older, from two retirement homes and a senior day care centre in the city of Pachuca, Mexico. Sociodemographic, socio-economic and behavioural variables were collected through a questionnaire. A trained and standardized examiner obtained the oral clinical variables. Chewing-stimulated saliva (paraffin method) was collected and the salivary flow rate, pH and buffer capacity were measured. The analysis was performed using non-parametric tests in Stata 9.0. Mean age was 79.1 ± 9.8 years. Most of the subjects included were women (69.1%). Mean chewing-stimulated salivary flow was 0.75 ± 0.80 mL/minute, and the pH and buffer capacity were 7.88 ± 0.83 and 4.20 ± 1.24, respectively. Mean chewing-stimulated salivary flow varied (p pH varied across the type of retirement home (p salivary features (salivary flow rate, pH and buffer capacity) and outline the variability of those features across selected sociodemographic, socio-economic and behavioural variables in a group of Mexican elders.

  4. Effect of sequence and stereochemistry reversal on p53 peptide mimicry.

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    Alessio Atzori

    Full Text Available Peptidomimetics effective in modulating protein-protein interactions and resistant to proteolysis have potential in therapeutic applications. An appealing yet underperforming peptidomimetic strategy is to employ D-amino acids and reversed sequences to mimic a lead peptide conformation, either separately or as the combined retro-inverso peptide. In this work, we examine the conformations of inverse, reverse and retro-inverso peptides of p53(15-29 using implicit solvent molecular dynamics simulation and circular dichroism spectroscopy. In order to obtain converged ensembles for the peptides, we find enhanced sampling is required via the replica exchange molecular dynamics method. From these replica exchange simulations, the D-peptide analogues of p53(15-29 result in a predominantly left-handed helical conformation. When the parent sequence is reversed sequence as either the L-peptide and D-peptide, these peptides display a greater helical propensity, feature reflected by NMR and CD studies in TFE/water solvent. The simulations also indicate that, while approximately similar orientations of the side-chains are possible by the peptide analogues, their ability to mimic the parent peptide is severely compromised by backbone orientation (for D-amino acids and side-chain orientation (for reversed sequences. A retro-inverso peptide is disadvantaged as a mimic in both aspects, and further chemical modification is required to enable this concept to be used fruitfully in peptidomimetic design. The replica exchange molecular simulation approach adopted here, with its ability to provide detailed conformational insights into modified peptides, has potential as a tool to guide structure-based design of new improved peptidomimetics.

  5. Bioorthogonal Diversification of Peptides through Selective Ruthenium(II)-Catalyzed C-H Activation.

    Science.gov (United States)

    Schischko, Alexandra; Ren, Hongjun; Kaplaneris, Nikolaos; Ackermann, Lutz

    2017-02-01

    Methods for the chemoselective modification of amino acids and peptides are powerful techniques in biomolecular chemistry. Among other applications, they enable the total synthesis of artificial peptides. In recent years, significant momentum has been gained by exploiting palladium-catalyzed cross-coupling for peptide modification. Despite major advances, the prefunctionalization elements on the coupling partners translate into undesired byproduct formation and lengthy synthetic operations. In sharp contrast, we herein illustrate the unprecedented use of versatile ruthenium(II)carboxylate catalysis for the step-economical late-stage diversification of α- and β-amino acids, as well as peptides, through chemo-selective C-H arylation under racemization-free reaction conditions. The ligand-accelerated C-H activation strategy proved water-tolerant and set the stage for direct fluorescence labelling as well as various modes of peptide ligation with excellent levels of positional selectivity in a bioorthogonal fashion. The synthetic utility of our approach is further demonstrated by twofold C-H arylations for the complexity-increasing assembly of artificial peptides within a multicatalytic C-H activation manifold. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Expression and Purification of C-Peptide Containing Insulin Using Pichia pastoris Expression System

    Directory of Open Access Journals (Sweden)

    Mohammed N. Baeshen

    2016-01-01

    Full Text Available Increase in the incidence of Insulin Dependent Diabetes Mellitus (IDDM among people from developed and developing countries has created a large global market for insulin. Moreover, exploration of new methods for insulin delivery including oral or inhalation route which require very high doses would further increase the demand of cost-effective recombinant insulin. Various bacterial and yeast strains have been optimized to overproduce important biopharmaceuticals. One of the approaches we have taken is the production of recombinant human insulin along with C-peptide in yeast Pichia pastoris. We procured a cDNA clone of insulin from Origene Inc., USA. Insulin cDNA was PCR amplified and cloned into yeast vector pPICZ-α. Cloned insulin cDNA was confirmed by restriction analysis and DNA sequencing. pPICZ-α-insulin clone was transformed into Pichia pastoris SuperMan5 strain. Several Zeocin resistant clones were obtained and integration of insulin cDNA in Pichia genome was confirmed by PCR using insulin specific primers. Expression of insulin in Pichia clones was confirmed by ELISA, SDS-PAGE, and Western blot analysis. In vivo efficacy studies in streptozotocin induced diabetic mice confirmed the activity of recombinant insulin. In conclusion, a biologically active human proinsulin along with C-peptide was expressed at high level using Pichia pastoris expression system.

  7. Levels of 25(OHD3, IL-2, and C-peptide in Children with Type 1 Diabetes Mellitus (T1DM Receiving Vitamin D3 Supplementation

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    Tjahyo Suryanto

    2018-01-01

    Full Text Available Type 1 Diabetes Mellitus (T1DM has become a health problem in many countries. T1DM is the consequence of autoimmune destruction process of β cells. There was relationship between vitamin D deficiency with T1DM. The destruction process was caused by an imbalance of pro-inflammatory and anti-inflammatory cytokines. One of the pro-inflammatory cytokines is IL-2. C-peptide examination to see the function of beta cells due to destruction of pancreatic beta cell. Administration of vitamin D3 supplementation still cause controversy and give varying results. This randomized clinical trial was conducted to determine the levels of 25(OHD3, IL-2, and C-peptide in people with T1DM who received vitamin D3 supplementation. The subjects were 26 children with T1DM, divided into K1 group (received vitamin D3 supplementation and K2 group (received placebo. The results showed higher levels of 25(OHD3 in the K1 group and statistically found a significant difference (p = 0.00. Higher levels of IL-2 and lower C-peptide were obtained in the K1 group and no statistically significant differences were found (p = 0.76 and p= 0.26. The insignificant relationship and the negative correlation were found between 25(OHD3 and IL-2 (p = 0.71; r = - 0.12, 25(OHD3 and C-peptide (p = 0.59; r = -0.16, also levels of IL-2 and C-peptide (p = 0.13; r = -0.44 in children with type 1 diabetes who received vitamin D3 supplementation. From this study can be concluded that administration vitamin D3 supplementation in patients with T1DM can increase levels 25(OHD3 significantly. This increase has not significantly lowered levels of IL-2 and increased levels of C-peptide. However, there was an absolute decrease in the rate of slower C-peptide in the supplemented group than in the placebo group.

  8. Isolation and Characterization of Antithrombin Peptides from the ...

    African Journals Online (AJOL)

    Purpose: To isolate and characterize the antithrombin compounds of Malaysian leeches' saliva collection (LSC) for use as anticoagulant proteins and peptides. Method: Reversed phase - high performance liquid chromatography (RP-HPLC) was used to isolate all proteins from LSC. All isolated proteins were tested for ...

  9. Factors determining the passage of drugs from blood into saliva.

    Science.gov (United States)

    Stephen, K W; McCrossan, J; Mackenzie, D; Macfarlane, C B; Speirs, C F

    1980-01-01

    1. Following single oral dosing of ampicillin, cephalexin, tetracycline, erythromycin estolate, clindamycin and rifampicin to six normal volunteers, antibacterial activity was measured at 1, 3 and 6 h in serum, gingival fluid and minor gland saliva from all subjects and in parotid and submandiabular saliva from three. 2. pH values of all gingival fluid and saliva specimens were noted. 3. Partition coefficients between n-octanol and water were measured for erythromycin, clindamycin and rifampicin. Published data were used for ampicillin, cephalexin and tetracycline. 4. All antibiotics, but particularly rifampicin, were detected in gingival fluid. Only rifampicin and to a lesser degree, clindamycin were present in the other salivary constituents. 5. In studies of secretion of drugs in saliva, both the physico-chemical characteristics of the drugs and the physiological differences between individual salivary components should be considered. 6. Parotid saliva samples are likely to be of greatest value. PMID:7356893

  10. Saliva and wound healing.

    Science.gov (United States)

    Brand, Henk S; Ligtenberg, Antoon J M; Veerman, Enno C I

    2014-01-01

    Oral wounds heal faster and with less scar formation than skin wounds. One of the key factors involved is saliva, which promotes wound healing in several ways. Saliva creates a humid environment, thus improving the survival and functioning of inflammatory cells that are crucial for wound healing. In addition, saliva contains several proteins which play a role in the different stages of wound healing. Saliva contains substantial amounts of tissue factor, which dramatically accelerates blood clotting. Subsequently, epidermal growth factor in saliva promotes the proliferation of epithelial cells. Secretory leucocyte protease inhibitor inhibits the tissue-degrading activity of enzymes like elastase and trypsin. Absence of this protease inhibitor delays oral wound healing. Salivary histatins in vitro promote wound closure by enhancing cell spreading and cell migration, but do not stimulate cell proliferation. A synthetic cyclic variant of histatin exhibits a 1,000-fold higher activity than linear histatin, which makes this cyclic variant a promising agent for the development of a new wound healing medication. Conclusively, recognition of the many roles salivary proteins play in wound healing makes saliva a promising source for the development of new drugs involved in tissue regeneration.

  11. Trefoil factors in saliva and gingival tissues of patients with chronic periodontitis

    DEFF Research Database (Denmark)

    Chaiyarit, Ponlatham; Chayasadom, Anek; Wara-Aswapati, Nawarat

    2012-01-01

    BACKGROUND: Trefoil factors (TFFs) are secreted molecules that are involved in cytoprotection against tissue damage and the immune response. TFFs have been detected in saliva and oral tissues, but their clinical significance has never been investigated in patients with chronic periodontitis....... The objective of this study is to determine whether TFF expression in saliva and gingival tissues is associated with periodontal pathology. METHODS: Saliva and gingival tissue samples were collected from 25 non-periodontitis individuals and 25 patients with chronic periodontitis (CP). Enzyme...... observed in patients with CP (P = 0.003 and P periodontal pathology and number of Porphyromonas gingivalis...

  12. Sensing pH via p-cyanophenylalanine fluorescence: Application to determine peptide pKa and membrane penetration kinetics.

    Science.gov (United States)

    Pazos, Ileana M; Ahmed, Ismail A; Berríos, Mariana I León; Gai, Feng

    2015-08-15

    We expand the spectroscopic utility of a well-known infrared and fluorescence probe, p-cyanophenylalanine, by showing that it can also serve as a pH sensor. This new application is based on the notion that the fluorescence quantum yield of this unnatural amino acid, when placed at or near the N-terminal end of a polypeptide, depends on the protonation status of the N-terminal amino group of the peptide. Using this pH sensor, we are able to determine the N-terminal pKa values of nine tripeptides and also the membrane penetration kinetics of a cell-penetrating peptide. Taken together, these examples demonstrate the applicability of using this unnatural amino acid fluorophore to study pH-dependent biological processes or events that accompany a pH change. Copyright © 2015 Elsevier Inc. All rights reserved.

  13. Effect of pH and fluoride on behavior of dental ZrO2 ceramics in artificial saliva

    International Nuclear Information System (INIS)

    Mukaeda, L.E.; Robin, A.; Santos, C.; Taguchi, S.P.; Borges Junior, L.A.

    2009-01-01

    A considerable increase in the ceramic products demand occurred due to the evolution of dental restoration techniques and these materials must resist to the complex mouth environment. The pH of saliva can decrease significantly due to the ingestion of acidic foods and beverages and mainly due to reactions occurring during bacteria metabolism that lead to the formation of organic acids. Fluorides are also present in the mouth since fluorides are usually added in drinking water, mouth washes, tooth pastes and gels for the prevention of plaque and caries formation. The combination of low pH and presence of fluorides can lead to the formation of HF and HF 2 - which are detrimental to metallic and probably to ceramic devices. In this work, commercial blocks of ZrO 2 ceramics (ProtMat Materiais Avancados® and Ivoclar®) were immersed in Fusayama artificial saliva of different pHs and fluoride concentrations. The properties of the as-produced ceramics (crystalline phases (XRD), microstructure (SEM), roughness (3D surface topography AFM) and mechanical resistance - Vickers hardness (Hv) and fracture toughness (KIC) were evaluated. Some of these properties were also determined after the immersion tests as well as the mass variation of the samples in order to evaluate the resistance of these ZrO 2 ceramics to degradation under these conditions. (author)

  14. Some Biological Activities of Malaysian Leech Saliva Extract

    OpenAIRE

    Abdualrahman M. Abdualkader; Ahmed Merzouk; Abbas Mohammed Ghawi; and Mohammed Alaama

    2011-01-01

    Leeches were fed on the phagostimulatory solution through parafilm membrane. The satiated leeches were forced to regurgitate the solution by soaking them in an ice-container. The anticoagulant activity was ascertained using thrombin time assay (TT). The result revealed that the saliva concentration which increases TT by 100% (IC100) is 43.205µg/ml plasma. The antimicrobial activity of the saliva was tested against several bacterial spp. (E.coli, P.aeruginosa, B.cereus, Sal.typhi and S...

  15. Saliva and dental erosion.

    Science.gov (United States)

    Buzalaf, Marília Afonso Rabelo; Hannas, Angélicas Reis; Kato, Melissa Thiemi

    2012-01-01

    Dental erosion is a multifactorial condition. The consideration of chemical, biological and behavioral factors is fundamental for its prevention and therapy. Among the biological factors, saliva is one of the most important parameters in the protection against erosive wear. This review discusses the role of salivary factors on the development of dental erosion. A search was undertaken on MeDLINe website for papers from 1969 to 2010. The keywords used in the research were "saliva", "acquired pellicle", "salivary flow", "salivary buffering capacity" and "dental erosion". Inclusion of studies, data extraction and quality assessment were undertaken independently and in duplicate by two members of the review team. Disagreements were solved by discussion and consensus or by a third party. Several characteristics and properties of saliva play an important role in dental erosion. Salivary clearance gradually eliminates the acids through swallowing and saliva presents buffering capacity causing neutralization and buffering of dietary acids. Salivary flow allows dilution of the acids. In addition, saliva is supersaturated with respect to tooth mineral, providing calcium, phosphate and fluoride necessary for remineralization after an erosive challenge. Furthermore, many proteins present in saliva and acquired pellicle play an important role in dental erosion. Saliva is the most important biological factor affecting the progression of dental erosion. Knowledge of its components and properties involved in this protective role can drive the development of preventive measures targeting to enhance its known beneficial effects.

  16. Saliva and dental erosion

    Directory of Open Access Journals (Sweden)

    Marília Afonso Rabelo Buzalaf

    2012-10-01

    Full Text Available Dental erosion is a multifactorial condition. The consideration of chemical, biological and behavioral factors is fundamental for its prevention and therapy. Among the biological factors, saliva is one of the most important parameters in the protection against erosive wear. Objective: This review discusses the role of salivary factors on the development of dental erosion. Material and Methods: A search was undertaken on MeDLINe website for papers from 1969 to 2010. The keywords used in the research were "saliva", "acquired pellicle", "salivary flow", "salivary buffering capacity" and "dental erosion". Inclusion of studies, data extraction and quality assessment were undertaken independently and in duplicate by two members of the review team. Disagreements were solved by discussion and consensus or by a third party. Results: Several characteristics and properties of saliva play an important role in dental erosion. Salivary clearance gradually eliminates the acids through swallowing and saliva presents buffering capacity causing neutralization and buffering of dietary acids. Salivary flow allows dilution of the acids. In addition, saliva is supersaturated with respect to tooth mineral, providing calcium, phosphate and fluoride necessary for remineralization after an erosive challenge. Furthermore, many proteins present in saliva and acquired pellicle play an important role in dental erosion. Conclusions: Saliva is the most important biological factor affecting the progression of dental erosion. Knowledge of its components and properties involved in this protective role can drive the development of preventive measures targeting to enhance its known beneficial effects.

  17. Circulating osteocrin stimulates bone growth by limiting C-type natriuretic peptide clearance.

    Science.gov (United States)

    Kanai, Yugo; Yasoda, Akihiro; Mori, Keita P; Watanabe-Takano, Haruko; Nagai-Okatani, Chiaki; Yamashita, Yui; Hirota, Keisho; Ueda, Yohei; Yamauchi, Ichiro; Kondo, Eri; Yamanaka, Shigeki; Sakane, Yoriko; Nakao, Kazumasa; Fujii, Toshihito; Yokoi, Hideki; Minamino, Naoto; Mukoyama, Masashi; Mochizuki, Naoki; Inagaki, Nobuya

    2017-11-01

    Although peptides are safe and useful as therapeutics, they are often easily degraded or metabolized. Dampening the clearance system for peptide ligands is a promising strategy for increasing the efficacy of peptide therapies. Natriuretic peptide receptor B (NPR-B) and its naturally occurring ligand, C-type natriuretic peptide (CNP), are potent stimulators of endochondral bone growth, and activating the CNP/NPR-B system is expected to be a powerful strategy for treating impaired skeletal growth. CNP is cleared by natriuretic peptide clearance receptor (NPR-C); therefore, we investigated the effect of reducing the rate of CNP clearance on skeletal growth by limiting the interaction between CNP and NPR-C. Specifically, we generated transgenic mice with increased circulating levels of osteocrin (OSTN) protein, a natural NPR-C ligand without natriuretic activity, and observed a dose-dependent skeletal overgrowth phenotype in these animals. Skeletal overgrowth in OSTN-transgenic mice was diminished in either CNP- or NPR-C-depleted backgrounds, confirming that CNP and NPR-C are indispensable for the bone growth-stimulating effect of OSTN. Interestingly, double-transgenic mice of CNP and OSTN had even higher levels of circulating CNP and additional increases in bone length, as compared with mice with elevated CNP alone. Together, these results support OSTN administration as an adjuvant agent for CNP therapy and provide a potential therapeutic approach for diseases with impaired skeletal growth.

  18. Different host complement systems and their interactions with saliva from Lutzomyia longipalpis (Diptera, Psychodidae and Leishmania infantum promastigotes.

    Directory of Open Access Journals (Sweden)

    Antonio Ferreira Mendes-Sousa

    Full Text Available BACKGROUND: Lutzomyia longipalpis is the vector of Leishmania infantum in the New World, and its saliva inhibits classical and alternative human complement system pathways. This inhibition is important in protecting the insect´s midgut from damage by the complement. L. longipalpis is a promiscuous blood feeder and must be protected against its host's complement. The objective of this study was to investigate the action of salivary complement inhibitors on the sera of different host species, such as dogs, guinea pigs, rats and chickens, at a pH of 7.4 (normal blood pH and 8.15 (the midgut pH immediately after a blood meal. We also investigated the role of the chicken complement system in Leishmania clearance in the presence and absence of vector saliva. RESULTS: The saliva was capable of inhibiting classical pathways in dogs, guinea pigs and rats at both pHs. The alternative pathway was not inhibited except in dogs at a pH of 8.15. The chicken classical pathway was inhibited only by high concentrations of saliva and it was better inhibited by the midgut contents of sand flies. Neither the saliva nor the midgut contents had any effect on the avian alternative pathway. Fowl sera killed L. infantum promastigotes, even at a low concentration (2%, and the addition of L. longipalpis saliva did not protect the parasites. The high body temperature of chickens (40°C had no effect on Leishmania viability during our assays. CONCLUSION: Salivary inhibitors act in a species-specific manner. It is important to determine their effects in the natural hosts of Leishmania infantum because they act on canid and rodent complements but not on chickens (which do not harbour the parasite. Moreover, we concluded that the avian complement system is the probable mechanism through which chickens eliminate Leishmania and that their high body temperature does not influence this parasite.

  19. Genetic selection of peptide aptamers that interact and inhibit both Small protein B and alternative ribosome-rescue factor A of Aeromonas veronii C4

    Directory of Open Access Journals (Sweden)

    Peng Liu

    2016-08-01

    Full Text Available Aeromonas veronii is a pathogenic gram-negative bacterium, which infects a variety of animals and results in mass mortality. The stalled-ribosome rescues are reported to ensure viability and virulence under stress conditions, of which primarily include trans-translation and alternative ribosome-rescue factor A (ArfA in A. veronii. For identification of specific peptides that interact and inhibit the stalled-ribosome rescues, peptide aptamer library (pTRG-SN-peptides was constructed using pTRG as vector and Staphylococcus aureus nuclease (SN as scaffold protein, in which 16 random amino acids were introduced to form an exposed surface loop. In the meantime both Small Protein B (SmpB which acts as one of the key components in trans-translation, and alternative ribosome-rescue factor A (ArfA were inserted to pBT to constitute pBT-SmpB and pBT-ArfA, respectively. The peptide aptamer PA-2 was selected from pTRG-SN-peptides by bacterial two-hybrid system (B2H employing pBT-SmpB or pBT-ArfA as baits. The conserved sites G133K134 and D138K139R140 of C-terminal SmpB were identified by interacting with N-terminal SN, and concurrently the residue K62 of ArfA was recognized by interacting with the surface loop of the specific peptide aptamer PA-2. The expression plasmids pN-SN or pN-PA-2, which combined the duplication origin of pRE112 with the neokanamycin promoter expressing SN or PA-2, were created and transformed into A. veronii C4, separately. The engineered A. veronii C4 which endowing SN or PA-2 expression impaired growth capabilities under stress conditions including temperatures, sucrose, glucose, potassium chloride (KCl and antibiotics, and the stress-related genes rpoS and nhaP were down-regulated significantly by Quantitative Real-time PCR (qRT-PCR when treating in 2.0% KCl. Thus,the engineered A. veronii C4 conferring PA-2 expression might be potentially attenuated vaccine, and also the peptide aptamer PA-2 could develop as anti

  20. Genetic Selection of Peptide Aptamers That Interact and Inhibit Both Small Protein B and Alternative Ribosome-Rescue Factor A of Aeromonas veronii C4.

    Science.gov (United States)

    Liu, Peng; Chen, Yong; Wang, Dan; Tang, Yanqiong; Tang, Hongqian; Song, Haichao; Sun, Qun; Zhang, Yueling; Liu, Zhu

    2016-01-01

    Aeromonas veronii is a pathogenic gram-negative bacterium, which infects a variety of animals and results in mass mortality. The stalled-ribosome rescues are reported to ensure viability and virulence under stress conditions, of which primarily include trans-translation and alternative ribosome-rescue factor A (ArfA) in A. veronii. For identification of specific peptides that interact and inhibit the stalled-ribosome rescues, peptide aptamer library (pTRG-SN-peptides) was constructed using pTRG as vector and Staphylococcus aureus nuclease (SN) as scaffold protein, in which 16 random amino acids were introduced to form an exposed surface loop. In the meantime both Small Protein B (SmpB) which acts as one of the key components in trans-translation, and ArfA were inserted to pBT to constitute pBT-SmpB and pBT-ArfA, respectively. The peptide aptamer PA-2 was selected from pTRG-SN-peptides by bacterial two-hybrid system (B2H) employing pBT-SmpB or pBT-ArfA as baits. The conserved sites G133K134 and D138K139R140 of C-terminal SmpB were identified by interacting with N-terminal SN, and concurrently the residue K62 of ArfA was recognized by interacting with the surface loop of the specific peptide aptamer PA-2. The expression plasmids pN-SN or pN-PA-2, which combined the duplication origin of pRE112 with the neokanamycin promoter expressing SN or PA-2, were created and transformed into A. veronii C4, separately. The engineered A. veronii C4 which endowing SN or PA-2 expression impaired growth capabilities under stress conditions including temperatures, sucrose, glucose, potassium chloride (KCl) and antibiotics, and the stress-related genes rpoS and nhaP were down-regulated significantly by Quantitative Real-time PCR (qRT-PCR) when treating in 2.0% KCl. Thus, the engineered A. veronii C4 conferring PA-2 expression might be potentially attenuated vaccine, and also the peptide aptamer PA-2 could develop as anti-microbial drugs targeted to the ribosome rescued factors in A

  1. Influence of saliva contamination on the shear bond strength of adhesives on enamel

    Directory of Open Access Journals (Sweden)

    Tatiana Feres Assad-Loss

    2012-04-01

    Full Text Available OBJECTIVE: To evaluate shear bond strength of 3 adhesive systems (Single Bond, TransbondTM MIP and TransbondTM XT applied on bovine enamel under saliva contamination condition. METHOD: One hundred and twenty enamel surfaces of bovine incisors were divided into 6 groups (n = 20 according to the adhesive system used (TransbondTM XT, TransbondTM MIP and Single Bond with or without saliva contamination. For each adhesive system, there were two groups defined as no contamination group (NC: 37% H3PO4 conditioning for 30 seconds and two layers of adhesive systems; saliva contamination group (SC: After the first adhesive layer application, the examined areas were contaminated with saliva. Samples were mounted appropriately for testing and stored in deionized water at 37 ºC for 7 days. Samples were then submitted to shear bond strength trials at a speed of 0.5 mm/min. The Adhesive Remnant Index (ARI was evaluated under stereomicroscopy. Two-way analysis of variance and the Tukey test were used to compare mean values (α = 0.05. RESULTS: Groups XT (NC = 26.29 ± 7.23; MIP (NC = 24.47 ± 7.52 and SB (NC = 32.36 ± 4.14 XT (SC = 19.59 ± 6.76; MIP (SC = 18.08 ± 6.39 and SB (SC = 18.18 ± 7.03 MPa. ARI 0 and 1 were the most prevalent scores in all study groups examined. CONCLUSION: Saliva contamination significantly decreased bond strength of the three adhesive systems examined (p <0.05. However, the comparison of groups with and without saliva contamination did not reveal any significant differences, and, therefore, the three systems may be considered equivalent.

  2. A Comparative Analysis of the Corrosive Effect of Artificial Saliva of Variable pH on DMLS and Cast Co-Cr-Mo Dental Alloy.

    Science.gov (United States)

    Puskar, Tatjana; Jevremovic, Danimir; Williams, Robert J; Eggbeer, Dominic; Vukelic, Djordje; Budak, Igor

    2014-09-11

    Dental alloys for direct metal laser sintering (DMLS) are available on the market today, but there is little scientific evidence reported on their characteristics. One of them is the release of ions, as an indicator of the corrosion characteristics of a dental alloy. Within this research, the difference in the elution of metals from DMLS and cast (CM) samples of Co-Cr-Mo dental alloy in saliva-like medium of three different pH was examined by inductively-coupled plasma mass spectrometry (ICP-MS). The obtained results show that the metal elution in artificial saliva from the DMLS alloy was lower than the elution from the CM alloy. The release of all investigated metal ions was influenced by the acidity, both from the DMLS and CM alloy, throughout the investigated period of 30 days. The change in acidity from a pH of 6.8 to a pH of 2.3 for the cast alloy led to a higher increase of the elution of Co, Cr and Mo from CM than from the DMLS alloy. The greatest release out of Co, Cr and Mo was for Co for both tested alloys. Further, the greatest release of all ions was measured at pH 2.3. In saliva of pH 2.3 and pH 4.5, the longer the investigated period, the higher the difference between the total metal ion release from the CM and DMLS alloys. Both alloys showed a safe level of elution according to the ISO definition in all investigated acidic environments.

  3. A Comparative Analysis of the Corrosive Effect of Artificial Saliva of Variable pH on DMLS and Cast Co-Cr-Mo Dental Alloy

    Directory of Open Access Journals (Sweden)

    Tatjana Puskar

    2014-09-01

    Full Text Available Dental alloys for direct metal laser sintering (DMLS are available on the market today, but there is little scientific evidence reported on their characteristics. One of them is the release of ions, as an indicator of the corrosion characteristics of a dental alloy. Within this research, the difference in the elution of metals from DMLS and cast (CM samples of Co-Cr-Mo dental alloy in saliva-like medium of three different pH was examined by inductively-coupled plasma mass spectrometry (ICP-MS. The obtained results show that the metal elution in artificial saliva from the DMLS alloy was lower than the elution from the CM alloy. The release of all investigated metal ions was influenced by the acidity, both from the DMLS and CM alloy, throughout the investigated period of 30 days. The change in acidity from a pH of 6.8 to a pH of 2.3 for the cast alloy led to a higher increase of the elution of Co, Cr and Mo from CM than from the DMLS alloy. The greatest release out of Co, Cr and Mo was for Co for both tested alloys. Further, the greatest release of all ions was measured at pH 2.3. In saliva of pH 2.3 and pH 4.5, the longer the investigated period, the higher the difference between the total metal ion release from the CM and DMLS alloys. Both alloys showed a safe level of elution according to the ISO definition in all investigated acidic environments.

  4. Saliva/serum ghrelin, obestatin and homocysteine levels in patients with ischaemic heart disease

    Science.gov (United States)

    Kilic, Nermin; Dagli, Necati; Aydin, Suleyman; Erman, Fazilet; Bek, Yuksel; Akin, Okhan; Kilic, SS; Erdemli, Haci Kemal; Alacam, Hasan

    2017-01-01

    Summary Background: We aimed to compare ghrelin, obestatin, homocysteine (Hcy), vitamin B12 and folate levels in the serum and saliva of ischaemic heart disease patients. Methods: Serum and saliva were collected from 33 ischaemic heart disease (IHD) patients and 28 age- and body mass index-matched healthy individuals. Levels of acylated and desacylated ghrelin, obestatin and Hcy were determined using the ELISA method. Results: Acylated ghrelin, desacylated ghrelin and obestatin levels in the saliva were found to be higher than those in the serum of the control group, while acylated and desacylated ghrelin levels in the saliva were significantly lower than those in the serum. Obestatin levels were higher in IHD patients (p = 0.001). Saliva and serum vitamin B12 and folate levels in IHD patients were significantly lower than in the control group (p = 0.001). Conclusions: It was determined that serum ghrelin levels increased in ischaemic heart disease patients, while serum levels of obestatin decreased. PMID:28759087

  5. Dynamic PET and SPECT imaging with radioiodinated, amyloid-reactive peptide p5 in mice: a positive role for peptide dehalogenation.

    Science.gov (United States)

    Martin, Emily B; Kennel, Stephen J; Richey, Tina; Wooliver, Craig; Osborne, Dustin; Williams, Angela; Stuckey, Alan; Wall, Jonathan S

    2014-10-01

    Dynamic molecular imaging provides bio-kinetic data that is used to characterize novel radiolabeled tracers for the detection of disease. Amyloidosis is a rare protein misfolding disease that can affect many organs. It is characterized by extracellular deposits composed principally of fibrillar proteins and hypersulfated proteoglycans. We have previously described a peptide, p5, which binds preferentially to amyloid deposits in a murine model of reactive (AA) amyloidosis. We have determined the whole body distribution of amyloid by molecular imaging techniques using radioiodinated p5. The loss of radioiodide from imaging probes due to enzymatic reaction has plagued the use of radioiodinated peptides and antibodies. Therefore, we studied iodine-124-labeled p5 by using dynamic PET imaging of both amyloid-laden and healthy mice to assess the rates of amyloid binding, the relevance of dehalogenation and the fate of the radiolabeled peptide. Rates of blood pool clearance, tissue accumulation and dehalogenation of the peptide were estimated from the images. Comparisons of these properties between the amyloid-laden and healthy mice provided kinetic profiles whose differences may prove to be indicative of the disease state. Additionally, we performed longitudinal SPECT/CT imaging with iodine-125-labeled p5 up to 72h post injection to determine the stability of the radioiodinated peptide when bound to the extracellular amyloid. Our data show that amyloid-associated peptide, in contrast to the unbound peptide, is resistant to dehalogenation resulting in enhanced amyloid-specific imaging. These data further support the utility of this peptide for detecting amyloidosis and monitoring potential therapeutic strategies in patients. Copyright © 2014 Elsevier Inc. All rights reserved.

  6. [Advances of tumor targeting peptides drug delivery system with pH-sensitive activities].

    Science.gov (United States)

    Ma, Yin-yun; Li, Li; Huang, Hai-feng; Gou, San-hu; Ni, Jing-man

    2016-05-01

    The pH-sensitive peptides drug delivery systems, which target to acidic extracellular environment of tumor tissue, have many advantages in drug delivery. They exhibit a high specificity to tumor and low cytotoxicity, which significantly increase the efficacy of traditional anti-cancer drugs. In recent years the systems have received a great attention. The pH-sensitive peptides drug delivery systems can be divided into five types according to the difference in pH-responsive mechanism,type of peptides and carrier materials. This paper summarizes the recent progresses in the field with a focus on the five types of pH-sensitive peptides in drug delivery systems. This may provide a guideline to design and application of tumor targeting drugs.

  7. A Convenient Approach to Synthesizing Peptide C-Terminal N-Alkyl Amides

    Science.gov (United States)

    Fang, Wei-Jie; Yakovleva, Tatyana; Aldrich, Jane V.

    2014-01-01

    Peptide C-terminal N-alkyl amides have gained more attention over the past decade due to their biological properties, including improved pharmacokinetic and pharmacodynamic profiles. However, the synthesis of this type of peptide on solid phase by current available methods can be challenging. Here we report a convenient method to synthesize peptide C-terminal N-alkyl amides using the well-known Fukuyama N-alkylation reaction on a standard resin commonly used for the synthesis of peptide C-terminal primary amides, the PAL-PEG-PS (Peptide Amide Linker-polyethylene glycol-polystyrene) resin. The alkylation and oNBS deprotection were conducted under basic conditions and were therefore compatible with this acid labile resin. The alkylation reaction was very efficient on this resin with a number of different alkyl iodides or bromides, and the synthesis of model enkephalin N-alkyl amide analogs using this method gave consistently high yields and purities, demonstrating the applicability of this methodology. The synthesis of N-alkyl amides was more difficult on a Rink amide resin, especially the coupling of the first amino acid to the N-alkyl amine, resulting in lower yields for loading the first amino acid onto the resin. This method can be widely applied in the synthesis of peptide N-alkyl amides. PMID:22252422

  8. Erosive potential of saliva stimulating tablets with and without fluoride in irradiated head and neck cancer patients

    International Nuclear Information System (INIS)

    Lajer, Christel; Buchwald, Christian; Nauntofte, Birgitte; Specht, Lena; Bardow, Allan; Jensdottir, Thorbjoerg

    2009-01-01

    Background: Patients irradiated in the head and neck region often suffer from severe dry mouth and use acidic saliva stimulating products, which may cause erosion of teeth. Purpose: To determine saliva stimulating effects and erosive potential (EP) of acidic saliva stimulating tablets (Xerodent TM ) with and without fluoride in irradiated head and neck cancer patients. Materials and method: Nineteen irradiated patients (median age 57 years) sucked Xerodent TM tablets with and without fluoride. Saliva collections were divided into three 10-min sessions in the sequence: unstimulated whole saliva, Xerodent TM stimulated saliva without fluoride, and with fluoride. Saliva pH was determined without loss of CO 2 and in combination with inorganic measures used to calculate the degree of saturation of hydroxyapatite (HAp) and fluorapatite (FAp). EP was determined directly in all saliva samples by monitored dissolution of HAp crystals. Results: Saliva flow rates increased significantly (15-fold) when sucking both tablets (p TM with and without fluoride were evaluated as non-erosive, however, for additional caries protection the fluoride variant is preferable.

  9. Radiation-induced parotid gland changes in oral cancer patients. Correlation between parotid volume and saliva production

    International Nuclear Information System (INIS)

    Teshima, Keiko; Murakami, Ryuji; Tomitaka, Etsuji

    2010-01-01

    The objective of this study was to evaluate whether saliva production reflects the parotid volume during the course of radiation therapy (RT) in patients with head-and-neck cancer. Twenty patients with advanced oral squamous cell carcinomas, who were treated with preoperative chemo-RT, underwent morphological assessment with CT or MRI and functional assessment with the Saxon test. For the Saxon test, saliva production was measured by weighing a gauze pad before and 2 min after chewing without swallowing; the low-normal value is 2 g. Saliva production and parotid volumes before and 2 weeks after RT were compared with the paired t-test, the Spearman rank correlation test and the Fisher exact test. After 30 Gy irradiation, mean saliva production was decreased from 4.2 to 1.0 g (P 3 (P<0.01); the post-RT: pre-RT parotid volume ratio ranged from 54% to 85% (mean 71%). Although the initial parotid volume was correlated with initial saliva production (r=0.47, P=0.04), no significant correlation was noted after RT (r=0.08, P=0.71), and there were considerable individual variations. The parotid volume ratio was inversely correlated with the saliva-reduction amount (r=-0.79, P<0.01). There was a correlation between decreased parotid gland volume and decreased saliva production in patients with head-and-neck cancer undergoing RT. Parotid volume reduction may predict parotid gland function. (author)

  10. Effect of Different Saliva Decontamination Procedures on Bond Strength to Dentin in Single Bottle Systems

    Directory of Open Access Journals (Sweden)

    M. Ghavam

    2004-09-01

    Full Text Available Statement of Problem: Following the increasing use of composites in restoring anterior and posterior teeth, problems due to its technique sensitivity have become a major concern.One of these problems is the possibility of contamination of dentin with saliva, blood and/or gingival fluid in different stages of bonding procedure, even with application of different methods of isolation. However, by introduction of Single-bottle dentin adhesives,the contamination possibility reduced to two stages. Scientific documents show that saliva contamination reduces bond strength of composites to dentin. Application of simple and efficient methods for reducing or eliminating saliva contamination enables clinicians to carry out dental treatment without any concern about deterioration of clinical longevity of restoration.Purpose: This study was designed to compare the effect of different decontamination methods on the shear bond strength of composite to dentin using a “Single-bottle” adhesive.Materials and Methods: Seventy-two extracted sound human molars and premolars were selected. Enamel of buccal surface was ground flat to expose dentin. The teeth were divided into 9 groups of 8 each. In control group (1 the adhesive “Excite” was used according tothe manufacturer, without any contamination. Conditioned and saliva contaminated dentin was (2 rinsed and blot dried, (3 rinsed, dried and re-etched. In groups 4, 5, 6 uncured adhesive was saliva contaminated and then: (4 only blot dried (5 rinsed, blot dried with adhesive reapplication and (6 resurfaced with bur, rinsed, dried and followed by repeating the whole process. In groups 7, 8, 9 cured adhesive was contaminated with saliva and then:(7 rinsed and dried (8 rinsed, blot dried with adhesive reapplication (9 same as group (6.Then “Tetric Ceram” composite cylinders were bonded to dentin surfaces. Samples were thermo cycled in 5°C and 55°C water, 30 seconds in each bath with a dowel time of 10

  11. Kinetics of Anti-Phlebotomus perniciosus Saliva Antibodies in Experimentally Bitten Mice and Rabbits.

    Directory of Open Access Journals (Sweden)

    Inés Martín-Martín

    Full Text Available Sand flies are hematophagous arthropods that act as vectors of Leishmania parasites. When hosts are bitten they develop cellular and humoral responses against sand fly saliva. A positive correlation has been observed between the number of bites and antibody levels indicating that anti-saliva antibody response can be used as marker of exposure to sand flies. Little is known about kinetics of antibodies against Phlebotomus perniciosus salivary gland homogenate (SGH or recombinant salivary proteins (rSP. This work focused on the study of anti-P. perniciosus saliva antibodies in sera of mice and rabbits that were experimentally exposed to the bites of uninfected sand flies.Anti-saliva antibodies were evaluated by ELISA and Western blot. In addition, antibody levels against two P. perniciosus rSP, apyrase rSP01B and D7 related protein rSP04 were determined in mice sera. Anti-saliva antibody levels increased along the immunizations and correlated with the number of sand fly bites. Anti-SGH antibody levels were detected in sera of mice five weeks after exposure, and persisted for at least three months. Anti-apyrase rSP01B antibodies followed similar kinetic responses than anti-SGH antibodies while rSP04 showed a delayed response and exhibited a greater variability among sera of immunized mice. In rabbits, anti-saliva antibodies appeared after the second week of exposure and IgG antibodies persisted at high levels, even 7 months post-exposure.Our results contributed to increase the knowledge on the type of immune response P. perniciosus saliva and individual proteins elicited highlighting the use of rSP01B as an epidemiological marker of exposure. Anti-saliva kinetics in sera of experimentally bitten rabbits were studied for the first time. Results with rabbit model provided useful information for a better understanding of the anti-saliva antibody levels found in wild leporids in the human leishmaniasis focus in the Madrid region, Spain.

  12. Structures of self-assembled amphiphilic peptide-heterodimers: effects of concentration, pH, temperature and ionic strength

    KAUST Repository

    Luo, Zhongli

    2010-01-01

    The amphiphilic double-tail peptides AXG were studied regarding secondary structure and self-assembly in aqueous solution. The two tails A = Ala 6 and G = Gly6 are connected by a central pair X of hydrophilic residues, X being two aspartic acids in ADG, two lysines in AKG and two arginines in ARG. The peptide AD (Ala6Asp) served as a single-tail reference. The secondary structure of the four peptides was characterized by circular dichroism spectroscopy under a wide range of peptide concentrations (0.01-0.8 mM), temperatures (20-98 °C), pHs (4-9.5) and ionic strengths. In salt-free water both ADG and AD form a β-sheet type of structure at high concentration, low pH and low temperature, in a peptide-peptide driven assembly of individual peptides. The transition has a two-state character for ADG but not for AD, which indicates that the added tail in ADG makes the assembly more cooperative. By comparison the secondary structures of AKG and ARG are comparatively stable over the large range of conditions covered. According to dynamic light scattering the two-tail peptides form supra-molecular aggregates in water, but high-resolution AFM-imaging indicate that ordered (self-assembled) structures are only formed when salt (0.1 M NaCl) is added. Since the CD-studies indicate that the NaCl has only a minor effect on the peptide secondary structure we propose that the main role of the added salt is to screen the electrostatic repulsion between the peptide building blocks. According to the AFM images ADG and AKG support a correlation between nanofibers and a β-sheet or unordered secondary structure, whereas ARG forms fibers in spite of lacking β-sheet structure. Since the AKG and ARG double-tail peptides self-assemble into distinct nanostructures while their secondary structures are resistant to environment factors, these new peptides show potential as robust building blocks for nano-materials in various medical and nanobiotechnical applications. © 2010 The Royal Society

  13. INFLUENCE OF SYSTEMIC DISEASES AND REMOVABLE ORTHODONTIC APPLIANCES ON THE QUALITY OF SALIVA IN CHILDHOOD.

    Directory of Open Access Journals (Sweden)

    Maya Rashkova

    2012-03-01

    Full Text Available During the last 10 years numerous investigations using saliva as a diagnostic tool have been carried out. The aim of present study is to evaluate saliva qualities for various general diseases and conditions that influence its qualities. (1 Evaluation of salivary flow and saliva consistency of children. (2 Evaluation of saliva pH and buffer capacity of children. Material and Methods. The investigation was carried out with 126 children (age 6 to 17 selected by their general diseases and conditions influencing the oral risk environment. The children were divided into 4 groups: 30 children with diabetes, 25 children with asthma treated with local corticosteroids, 27 healthy children with orthodontic treatment, 34 children as a control group (healthy children. The saliva of the children was tested with the help of “Saliva Check” of GC company. The instructions of the company producer were followed.Results. Stimulated saliva current is reliably lower for children with asthma treated with local corticosteroids, diabetes and children with orthodontic appliances. Saliva pH is with lower values for children with diabetes and asthma – diseases predisposing to acid oral environment. The decreased saliva buffer capacity for children with diabetes and asthma is an indicator for the difficult regulation of the dynamically changing oral electrolytic balance of those children.Conclusion. The saliva parameters studied can be used as biomarkers of the liquid oral environment with regard to the risks for caries and periodontal diseases in children. General health status influences saliva qualities increasing thus indirectly the caries risk.

  14. Markers in blood and saliva for prediction of orthodontically induced inflammatory root resorption: a retrospective case controlled-study

    Directory of Open Access Journals (Sweden)

    Dilara Yashin

    2017-09-01

    Full Text Available Abstract Background Hormonal and enzymatic factors may render certain individuals more susceptible to orthodontically induced inflammatory root resorption (OIIRR. The objectives of this study are (1 to identify biochemical key markers in blood and saliva that may be correlated to the trend of extensive OIIRR and (2 to utilise these markers to predict a susceptible patient-receiving orthodontic treatment. Methods Nine patients (mean age 23 + 2.9 years who had moderate to severe OIIRR that assessed via orthopantomograms and met the inclusion criteria were classified as the root resorption group (RRG. Blood chemistry was evaluated using the collection of fasting blood and unstimulated saliva samples. Multiplex enzyme-linked immunosorbent assay (ELISA arrays were used to screen blood and saliva samples for human cytokines, chemokines and several key enzymes that may play a role in root resorption following orthodontic force application. Biochemical findings from 16 matching subjects were used as the control (CG for comparative measurements. Results Patients with moderate to severe OIIRR showed a significant increase in salivary cytokines including interleukin (IL 7, IL-10, IL-12p70 and interferon-gamma (IFN-γ level as well as a significant decrease in IL-4 level. Osteocalcin and procollagen type I N-terminal peptide (P1NP appeared to be the only blood factors that showed a significant difference, more in the CG than the RRG. Conclusions Saliva might be a more valuable way of measuring changes in cytokine expression than blood secondary to orthodontic treatment. Although the increased expression of pro-inflammatory and anti-inflammatory cytokines may be determinants in the development of moderate to severe OIIRR, cytokine expression may be affected by several potential inflammations in another part of the body. Future research could investigate the cause/effect relationship of different cytokines, in a larger group of patients and at different

  15. Continuous analysis of parotid saliva during resting and short-duration simulated chewing

    NARCIS (Netherlands)

    Neyraud, E.; Bult, J.H.F.; Dransfield, E.

    2009-01-01

    Objective: Parotid saliva flow is increased by mastication and its composition is also modified. The aim of this work was to clarify the relationships between flow rate, pH and protein concentration, during resting and short-duration simulated chewing, using continuous and fractional saliva

  16. Amyloid fibril formation of peptides derived from the C-terminus of CETP modulated by lipids

    Energy Technology Data Exchange (ETDEWEB)

    García-González, Victor [Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, 04510 México, DF (Mexico); Mas-Oliva, Jaime, E-mail: jmas@ifc.unam.mx [Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, 04510 México, DF (Mexico); División de Investigación, Facultad de Medicina, Universidad Nacional Autónoma de México, 04510 México, DF (Mexico)

    2013-04-26

    Highlights: •The secondary structure of a C-terminal peptide derived from CETP was studied. •Lipids modulate secondary structure changes of a C-terminal peptide derived from CETP. •Lysophosphatidic acid maintains a functional α-helix and prevents fibril formation. •Transfer of lipids by CETP is related to the presence of an α-helix at its C-end. -- Abstract: Cholesteryl-ester transfer protein (CETP) is a plasmatic protein involved in neutral lipid transfer between lipoproteins. Focusing on the last 12 C-terminus residues we have previously shown that mutation D{sub 470}N promotes a conformational change towards a β-secondary structure. In turn, this modification leads to the formation of oligomers and fibrillar structures, which cause cytotoxic effects similar to the ones provoked by amyloid peptides. In this study, we evaluated the role of specific lipid arrangements on the structure of peptide helix-Z (D{sub 470}N) through the use of thioflavin T fluorescence, peptide bond absorbance, circular dichroism and electron microscopy. The results indicate that the use of micelles formed with lysophosphatidylcholine and lysophosphatidic acid (LPA) under neutral pH induce a conformational transition of peptide helix-Z containing a β-sheet conformation to a native α-helix structure, therefore avoiding the formation of amyloid fibrils. In contrast, incubation with phosphatidic acid does not change the profile for the β-sheet conformation. When the electrostatic charge at the surface of micelles or vesicles is regulated through the use of lipids such as phospholipid and LPA, minimal changes and the presence of β-structures were recorded. Mixtures with a positive net charge diminished the percentage of β-structure and the amount of amyloid fibrils. Our results suggest that the degree of solvation determined by the presence of a free hydroxyl group on lipids such as LPA is a key condition that can modulate the secondary structure and the consequent formation of

  17. Amyloid fibril formation of peptides derived from the C-terminus of CETP modulated by lipids

    International Nuclear Information System (INIS)

    García-González, Victor; Mas-Oliva, Jaime

    2013-01-01

    Highlights: •The secondary structure of a C-terminal peptide derived from CETP was studied. •Lipids modulate secondary structure changes of a C-terminal peptide derived from CETP. •Lysophosphatidic acid maintains a functional α-helix and prevents fibril formation. •Transfer of lipids by CETP is related to the presence of an α-helix at its C-end. -- Abstract: Cholesteryl-ester transfer protein (CETP) is a plasmatic protein involved in neutral lipid transfer between lipoproteins. Focusing on the last 12 C-terminus residues we have previously shown that mutation D 470 N promotes a conformational change towards a β-secondary structure. In turn, this modification leads to the formation of oligomers and fibrillar structures, which cause cytotoxic effects similar to the ones provoked by amyloid peptides. In this study, we evaluated the role of specific lipid arrangements on the structure of peptide helix-Z (D 470 N) through the use of thioflavin T fluorescence, peptide bond absorbance, circular dichroism and electron microscopy. The results indicate that the use of micelles formed with lysophosphatidylcholine and lysophosphatidic acid (LPA) under neutral pH induce a conformational transition of peptide helix-Z containing a β-sheet conformation to a native α-helix structure, therefore avoiding the formation of amyloid fibrils. In contrast, incubation with phosphatidic acid does not change the profile for the β-sheet conformation. When the electrostatic charge at the surface of micelles or vesicles is regulated through the use of lipids such as phospholipid and LPA, minimal changes and the presence of β-structures were recorded. Mixtures with a positive net charge diminished the percentage of β-structure and the amount of amyloid fibrils. Our results suggest that the degree of solvation determined by the presence of a free hydroxyl group on lipids such as LPA is a key condition that can modulate the secondary structure and the consequent formation of amyloid

  18. Protein components in saliva and plaque fluid from irradiated primates

    Energy Technology Data Exchange (ETDEWEB)

    Edgar, W.M.; Bowen, W.H.; Cole, M.F. (Caries Prevention and Research Branch, National Caries Program, NIDR, Bethesda, Maryland, USA)

    1982-01-01

    Irradiation of the major salivary glands of monkeys (Macaca mulatta) fed cariogenic diets leads to caries clinically indistinguishable from radiation caries in man. This study compares the organic compostion of individual samples of plaque fluid and saliva from irradiated and control monkeys receiving the same cariogenic diet. Plaque and saliva were collected from fasting, tranquillised animals. Four irradiated animals were sampled repeatedly as were non-irradiated controls. Total protein, albumin, immunoglobulins A, G, and M, and the third component of complement (C'3) were quantitated in plaque fluid and whole saliva. Salivary amylase and peroxidase activities were also determined. Plaque fluid and saliva samples were also subjected to polyacrylamide gel electrophoresis. The total viable anaerobic count and numbers of Streptococcus mutans were determined in samples of plaque. The results suggest that the major effect of irradiation leading to increased numbers of S. mutans and caries susceptibility is in the amount, and not the composition, of the saliva produced by the residual gland tissue. The scanty flow of saliva may reduce the effectiveness of cleansing, buffering and lubrication mechanisms as well as resulting in a marked reduction in the total amount of specific and non-specific immune factors entering the mouth.

  19. Protein components in saliva and plaque fluid from irradiated primates

    International Nuclear Information System (INIS)

    Edgar, W.M.; Bowen, W.H.; Cole, M.F.

    1982-01-01

    Irradiation of the major salivary glands of monkeys (Macaca mulatta) fed cariogenic diets leads to caries clinically indistinguishable from radiation caries in man. This study compares the organic compostion of individual samples of plaque fluid and saliva from irradiated and control monkeys receiving the same cariogenic diet. Plaque and saliva were collected from fasting, tranquillised animals. Four irradiated animals were sampled repeatedly as were non-irradiated controls. Total protein, albumin, immunoglobulins A, G, and M, and the third component of complement (C'3) were quantitated in plaque fluid and whole saliva. Salivary amylase and peroxidase activities were also determined. Plaque fluid and saliva samples were also subjected to polyacrylamide gel electrophoresis. The total viable anaerobic count and numbers of Streptococcus mutans were determined in samples of plaque. The results suggest that the major effect of irradiation leading to increased numbers of S. mutans and caries susceptibility is in the amount, and not the composition, of the saliva produced by the residual gland tissue. The scanty flow of saliva may reduce the effectiveness of cleansing, buffering and lubrication mechanisms as well as resulting in a marked reduction in the total amount of specific and non-specific immune factors entering the mouth. (author)

  20. Protein components in saliva and plaque fluid from irradiated primates

    Energy Technology Data Exchange (ETDEWEB)

    Edgar, W M; Bowen, W H; Cole, M F [Caries Prevention and Research Branch, National Caries Program, NIDR, Bethesda, Maryland, USA

    1982-01-01

    Irradiation of the major salivary glands of monkeys (Macaca mulatta) fed cariogenic diets leads to caries clinically indistinguishable from radiation caries in man. This study compares the organic compostion of individual samples of plaque fluid and saliva from irradiated and control monkeys receiving the same cariogenic diet. Plaque and saliva were collected from fasting, tranquillised animals. Four irradiated animals were sampled repeatedly as were non-irradiated controls. Total protein, albumin, immunoglobulins A, G, and M, and the third component of complement (C'3) were quantitated in plaque fluid and whole saliva. Salivary amylase and peroxidase activities were also determined. Plaque fluid and saliva samples were also subjected to polyacrylamide gel electrophoresis. The total viable anaerobic count and numbers of Streptococcus mutans were determined in samples of plaque. The results suggest that the major effect of irradiation leading to increased numbers of S. mutans and caries susceptibility is in the amount, and not the composition, of the saliva produced by the residual gland tissue. The scanty flow of saliva may reduce the effectiveness of cleansing, buffering and lubrication mechanisms as well as resulting in a marked reduction in the total amount of specific and non-specific immune factors entering the mouth.

  1. Improving ambulatory saliva-sampling compliance in pregnant women: a randomized controlled study.

    Directory of Open Access Journals (Sweden)

    Julian Moeller

    Full Text Available OBJECTIVE: Noncompliance with scheduled ambulatory saliva sampling is common and has been associated with biased cortisol estimates in nonpregnant subjects. This study is the first to investigate in pregnant women strategies to improve ambulatory saliva-sampling compliance, and the association between sampling noncompliance and saliva cortisol estimates. METHODS: We instructed 64 pregnant women to collect eight scheduled saliva samples on two consecutive days each. Objective compliance with scheduled sampling times was assessed with a Medication Event Monitoring System and self-reported compliance with a paper-and-pencil diary. In a randomized controlled study, we estimated whether a disclosure intervention (informing women about objective compliance monitoring and a reminder intervention (use of acoustical reminders improved compliance. A mixed model analysis was used to estimate associations between women's objective compliance and their diurnal cortisol profiles, and between deviation from scheduled sampling and the cortisol concentration measured in the related sample. RESULTS: Self-reported compliance with a saliva-sampling protocol was 91%, and objective compliance was 70%. The disclosure intervention was associated with improved objective compliance (informed: 81%, noninformed: 60%, F(1,60  = 17.64, p<0.001, but not the reminder intervention (reminders: 68%, without reminders: 72%, F(1,60 = 0.78, p = 0.379. Furthermore, a woman's increased objective compliance was associated with a higher diurnal cortisol profile, F(2,64  = 8.22, p<0.001. Altered cortisol levels were observed in less objective compliant samples, F(1,705  = 7.38, p = 0.007, with delayed sampling associated with lower cortisol levels. CONCLUSIONS: The results suggest that in pregnant women, objective noncompliance with scheduled ambulatory saliva sampling is common and is associated with biased cortisol estimates. To improve sampling compliance, results suggest

  2. Effect of pH and fluoride on behavior of dental ZrO{sub 2} ceramics in artificial saliva

    Energy Technology Data Exchange (ETDEWEB)

    Mukaeda, L.E.; Robin, A.; Santos, C.; Taguchi, S.P.; Borges Junior, L.A., E-mail: luizamukaeda@gmail.com, E-mail: alain@demar.eel.usp.br, E-mail: claudinei@demar.eel.usp.br, E-mail: simone@demar.eel.usp.br, E-mail: borges.jr@itelefonica.com.br [Universidade de Sao Paulo (EEL/DEMAR/USP), Lorena, SP (Brazil). Escola de Engenharia; Machado, J.P.B., E-mail: joaopaulo@las.inpe.br [Instituto Nacional de Pesquisas Espaciais (INPE), Sao Jose dos Campos, SP (Brazil)

    2009-07-01

    A considerable increase in the ceramic products demand occurred due to the evolution of dental restoration techniques and these materials must resist to the complex mouth environment. The pH of saliva can decrease significantly due to the ingestion of acidic foods and beverages and mainly due to reactions occurring during bacteria metabolism that lead to the formation of organic acids. Fluorides are also present in the mouth since fluorides are usually added in drinking water, mouth washes, tooth pastes and gels for the prevention of plaque and caries formation. The combination of low pH and presence of fluorides can lead to the formation of HF and HF{sub 2}{sup -} which are detrimental to metallic and probably to ceramic devices. In this work, commercial blocks of ZrO{sub 2} ceramics (ProtMat Materiais Avancados® and Ivoclar®) were immersed in Fusayama artificial saliva of different pHs and fluoride concentrations. The properties of the as-produced ceramics (crystalline phases (XRD), microstructure (SEM), roughness (3D surface topography AFM) and mechanical resistance - Vickers hardness (Hv) and fracture toughness (KIC) were evaluated. Some of these properties were also determined after the immersion tests as well as the mass variation of the samples in order to evaluate the resistance of these ZrO{sub 2} ceramics to degradation under these conditions. (author)

  3. Short- to medium-term effects of consumption of quebracho tannins on saliva production and composition in sheep and goats.

    Science.gov (United States)

    Salem, A Z M; López, S; Ranilla, M J; González, J S

    2013-03-01

    Eight Merino sheep (49.4 ± 4.23 kg BW) and 8 Alpine goats (53.2 ± 2.51 kg BW) were used to study the effect of ingestion of quebracho tannins on salivation. Four sheep and 4 goats were individually fed a daily allotment of 20 g DM of alfalfa hay/kg BW (Control). Another 4 sheep and 4 goats were also given 20 g DM of alfalfa hay/kg BW supplemented with 50 g of quebracho/kg DM (Tannin) for a period of 64 d. The saliva secretion from the left parotid gland was collected by insertion of a polyvinyl chloride catheter into the parotid duct and the amount of parotid saliva produced recorded over three 48-h periods on d 1 and 2 (P1), d 31 and 32 (P2), and d 61 and 62 (P3) after the tannin feeding was initiated. The total amount of saliva produced was estimated from rumen water kinetics determined on d 4, d 34, and d 64 of the experiment. Experimental design was completely randomized, with repeated measures on each experimental unit, performing separate analysis for sheep and goats. Parotid saliva production was not affected by the sampling period in either animal species receiving the Control diet. Corresponding values for sheep were 2.04, 2.12, and 2.27 L/d (P = 0.89) and for goats 1.65, 1.79, and 1.86 L/d (P = 0.95). Sheep fed the Tannin diet produced 55, 73, and 107% of the amount of saliva recorded in sheep fed the Control diet on P1, P2, or P3, respectively. Corresponding values in goats were 88, 130, and 134% on P1, P2, or P3, respectively. Estimated total saliva production was not affected (P = 0.50 for sheep and P = 0.97 for goats) by the ingestion of quebracho. There was no difference (P > 0.10) in osmotic pressure, P, Mg, Ca, urea, and protein concentrations in parotid saliva. There were, however, differences in Na and K concentrations in response to the ingestion of quebracho tannins, with Na concentrations increasing (P = 0.05) and K concentrations decreasing (P = 0.04) in sheep saliva and pH increasing (P = 0.05) in goat saliva. In conclusion, the inclusion

  4. Lysine as helix C-capping residue in a synthetic peptide.

    Science.gov (United States)

    Esposito, G; Dhanapal, B; Dumy, P; Varma, V; Mutter, M; Bodenhausen, G

    1997-01-01

    The structure of the synthetic peptide CH3CO(Leu-Ser-Leu-Leu-Leu-Ser-Leu)3Lys-NH2 in trifluoroethanol/water 60/40 (volume ratio) was characterized by two-dimensional nmr spectroscopy. The peptide, closely related to the amphiphilic helix models designed by W. F. De-Grado and co-workers to mimic protein ion channels [(1988) Science, Vol. 240, p. 1177-1181], folds into a regular helix spanning residues 1-20. Evidence for a helix C-terminal capping conformation, involving the terminal lysine residue, was observed from Overhauser effects and checked for consistency by restrained molecular dynamics simulations. The side-chain amino group of Lys22 forms a hydrogen bond with the carbonyl of Leu18, and the distorted helical geometry of the terminal dipeptide allows the inclusion of a water bridge between the backbone NH of the Lys22 residue and the carbonyls of Leu19 and Ser20.

  5. Oestrogens in saliva

    International Nuclear Information System (INIS)

    Rothschild, R.S.; Levine, L.S.; Hattingh, J.

    1981-01-01

    Matched plasma and saliva samples were obtained from a non-pregnant and pregnant group (last trimester) of female caucasians. Using a sensitive radioimmunoassay, 17β-oestradiol was measured, and the gingival index system of Loe (1967) was used to assess the gingival condition of each patient. The results showed that 17β-oestradiol could be measured in saliva but that the levels were extremely low and a very sensitive immunoassay was necessary. In the pregnant group, saliva represented 3 per cent of the plasma level. This was not the case in the non-pregnant group, probably because of the constantly changing free: bound plasma ratio. The results are discussed in relation to the fact that oestrogens are known to bind to the oral epithelium [af

  6. Fluoride kinetics in saliva after the use of a fluoride-containing chewing gum Cinética de flúor na saliva após o uso de uma goma de mascar fluoretada

    Directory of Open Access Journals (Sweden)

    Maria Francisca Thereza Borro Bijella

    2005-12-01

    Full Text Available There is a relationship between the use of fluoride, the reduction of dental caries and the increase of dental fluorosis. The purpose of this study was to analyze the fluoride kinetics in saliva after using the HappydentTM chewing gum, which contains 3.38 mg of fluoride as monofluorophosphate. Fifteen 7-9-year-old volunteers were instructed to chew the gum TridentTM (control and HappydentTM on different days. Total saliva was collected for 3 minutes, at 0, 3, 6, 9, 15, 30 and 45 minutes after starting chewing. Salivary fluoride was analyzed with a fluoride-specific electrode (Orion 96-09 after acid hydrolysis. The data were analyzed by two-way analysis of variance and by Tukey’s post hoc test (p Há uma relação entre o uso de fluoretos, a redução na cárie e o aumento da fluorose dentária. O objetivo deste estudo foi analisar a cinética do flúor na saliva após o uso da goma de mascar Happydent®, que contém 3,38 mg de flúor como monofluorfosfato. A saliva foi coletada de 15 voluntários entre 7 e 9 anos de idade, durante 3 minutos nos intervalos de 0, 3, 6, 9, 15, 30 e 45 minutos. Inicialmente, a coleta foi realizada com o Trident® (controle e, após 24 h, a coleta foi repetida com a goma de mascar Happydent®. O flúor foi analisado com um eletrodo íon-específico (Orion 96-09 após a realização da hidrólise ácida. Os dados foram analisados através da análise de variância a dois critérios e pelo teste de Tukey (p < 0,05. A quantidade média ± dp (mg de flúor liberado na saliva foi 0,276 ± 0,126 e 0,024 ± 0,014 para o Happydent® e o Trident®, respectivamente. A quantidade de flúor nas amostras de saliva após o uso do Happydent® foi significativamente maior do que após o uso do Trident® em todos os tempos experimentais, com exceção dos períodos de 30 e 45 minutos. A alta quantidade de flúor na saliva após o uso do Happydent® poderia ser eficiente na prevenção da cárie dentária, o que deveria ser avaliado

  7. IgE and IgG cross-reactivity among Lol p I and Lol p II/III. Identification of the C-termini of Lol p I, II, and III as cross-reactive structures

    NARCIS (Netherlands)

    van Ree, R.; van Leeuwen, W. A.; van den Berg, M.; Weller, H. H.; Aalberse, R. C.

    1994-01-01

    In this study, the homologous C-termini of Lol p I, Lol p II, and Lol p III were shown to contain cross-reactive B-cell epitopes. This was demonstrated by inhibition studies with purified Lol p I, II, and III and synthetic peptides of their C-termini. It was ruled out that the observed

  8. Reversibly Switchable, pH-Dependent Peptide Ligand Binding via 3,5-Diiodotyrosine Substitutions.

    Science.gov (United States)

    Ngambenjawong, Chayanon; Sylvestre, Meilyn; Gustafson, Heather H; Pineda, Julio Marco B; Pun, Suzie H

    2018-04-20

    Cell type-specific targeting ligands utilized in drug delivery applications typically recognize receptors that are overexpressed on the cells of interest. Nonetheless, these receptors may also be expressed, to varying extents, on off-target cells, contributing to unintended side effects. For the selectivity profile of targeting ligands in cancer therapy to be improved, stimuli-responsive masking of these ligands with acid-, redox-, or enzyme-cleavable molecules has been reported, whereby the targeting ligands are exposed in specific environments, e.g., acidic tumor hypoxia. One possible drawback of these systems lies in their one-time, permanent trigger, which enables the "demasked" ligands to bind off-target cells if released back into the systemic circulation. A promising strategy to address the aforementioned problem is to design ligands that show selective binding based on ionization state, which may be microenvironment-dependent. In this study, we report a systematic strategy to engineer low pH-selective targeting peptides using an M2 macrophage-targeting peptide (M2pep) as an example. 3,5-Diiodotyrosine mutagenesis into native tyrosine residues of M2pep confers pH-dependent binding behavior specific to acidic environment (pH 6) when the amino acid is protonated into the native tyrosine-like state. At physiological pH of 7.4, the hydroxyl group of 3,5-diiodotyrosine on the peptide is deprotonated leading to interruption of the peptide native binding property. Our engineered pH-responsive M2pep (Ac-Y-Î-Î) binds target M2 macrophages more selectively at pH 6 than at pH 7.4. In addition, 3,5-diiodotyrosine substitutions also improve serum stability of the peptide. Finally, we demonstrate pH-dependent reversibility in target binding via a postbinding peptide elution study. The strategy presented here should be applicable for engineering pH-dependent functionality of other targeting peptides with potential applications in physiology-dependent in vivo targeting

  9. Phase behavior and nanoscale structure of phospholipid membranes incorporated with acylated C-14-peptides

    DEFF Research Database (Denmark)

    Pedersen, T.B.; Kaasgaard, Thomas; Jensen, M.O.

    2005-01-01

    The thermotropic phase behavior and lateral structure of dipalmitoylphosphatidylcholine (DPPC) lipid bilayers containing an acylated peptide has been characterized by differential scanning calorimetry (DSC) on vesicles and atomic force microscopy (AFM) on mica-supported bilayers. The acylated...... peptide, which is a synthetic decapeptide N-terminally linked to a C-14 acyl chain (C-14-peptide), is incorporated into DPPC bilayers in amounts ranging from 0-20 mol %. The calorimetric scans of the two-component system demonstrate a distinct influence of the C-14-peptide on the lipid bilayer...... gel phase DPPC bilayers, inserts preferentially into preexisting defect regions and has a noticeable influence on the organization of the surrounding lipids. The presence of the C-14-peptide gives rise to a laterally heterogeneous bilayer structure with coexisting lipid domains characterized by a 10...

  10. Pengaruh Transplantasi Allograf Pancreatic Stem Cell terhadap Kadar Insulin dan C-Peptide Tikus Putih Penderita Diabetes Melitus Tipe I

    Directory of Open Access Journals (Sweden)

    Boedi Setiawan

    2016-09-01

    Full Text Available Diabetes mellitus is one of the degenerative diseases in which the therapy still remains unresolved and is still a serious threat to the global health, including to the health of Indonesian people. The aim of this study was to describe the level of insulin and C-peptide in diabetes mellitus type I white rats treated with pancreatic stem cell allograft through intrapancreatic laparotomy. This study was conducted at the Institute of Tropical Diseases, Universitas Airlangga, Surabaya in a 6 month period (July–December 2014. Twelve male white rats Rattus novergicus Wistar strain, were randomly divided into two groups. The first group (P0 was injected by alloxan, 150 mg/kg body weight, without stem cell therapy. Another group was injected by alloxan, 150 mg/kg body weight, and was treated with 1x106/kg body weight pancreatic stem cell throughintrapancreatic laparotomy (P1. The experiment was finalized on the 31th day of the experiment. The results showed that the blood glucose levels at the end of experiment were highly significantly different p<0.01 between the treatment group that received stem cell therapy (P1 and P0 positive control, although the average value of blood glucose levels was not as normal as on the first day. C-peptide and insulin levels of P0 and P1 group differed significantly (p<0.01. It can be concluded that stem cell therapy through intrapancreatic laparotomy can reduce blood glucose levels and increase the levels of C-peptide and insulin.

  11. Preliminary findings on the correlation of saliva pH, buffering capacity, flow rate and consistency in relation to waterpipe tobacco smoking.

    Science.gov (United States)

    Khemiss, Mehdi; Ben Khelifa, Mohamed; Ben Saad, Helmi

    2017-12-01

    The aim of the present comparative study was to compare some salivary characteristics between exclusive waterpipe smokers (EWPS) and non-smokers. 72 males (36 EWPS) were recruited. The volume of stimulated saliva was determined and divided by the duration of saliva collection. The pH was measured directly using a pH meter. The buffering capacity was determined using a quantitative method which involved the addition of 10 µl HCl. Up to a total of 160 µL was titrated up to obtain a pH titration curve. At 50 µL of titrated HCl, buffering capacity was ranked into three categories: high, medium and low. EWPS and non-smoker groups had similar flow rates (1.81 ± 0.79 and 1.78 ± 1.14 mL min-1) and similar baseline pH (6.60 ± 0.37 and 6.76 ± 0.39). Statistically significant differences in the two groups' pH were observed from 30 to 160 µL of titrated up HCl. At 50 µL of titrated up HCl, the EWPS group compared to the non-smoker group had a significantly higher pH (4.79 ± 0.72 vs. 5.32 ± 0.79). To conclude, waterpipe tobacco smoking alters the buffering capacity but does not alter either salivary flow rates or the baseline pH and consistency.

  12. Electrochemical Behaviour and Galvanic Effects of Titanium Implants Coupled to Metallic Suprastructures in Artificial Saliva

    Science.gov (United States)

    Mellado-Valero, Ana; Igual Muñoz, Anna; Guiñón Pina, Virginia

    2018-01-01

    The aim of the present study is to analyze the electrochemical behavior of five different dental alloys: two cobalt-chromium alloys (CoCr and CoCr-c), one nickel-chromium-titanium alloy (NiCrTi), one gold-palladium alloy (Au), and one titanium alloy (Ti6Al4V), and the galvanic effect when they are coupled to titanium implants (TiG2). It was carried out by electrochemical techniques (open circuit measurements, potentiodynamic curves and Zero-Resistance Ammetry) in artificial saliva (AS), with and without fluorides in different acidic conditions. The studied alloys are spontaneously passivated, but NiCrTi alloy has a very narrow passive domain and losses its passivity in presence of fluorides, so is not considered as a good option for implant superstructures. Variations of pH from 6.5 to 3 in artificial saliva do not change the electrochemical behavior of Ti, Ti6Al4V, and CoCr alloys, and couples, but when the pH of the artificial saliva is below 3.5 and the fluoride content is 1000 ppm Ti and Ti6Al4V starts actively dissolving, and CoCr-c superstructures coupled to Ti show acceleration of corrosion due to galvanic effects. Thus, NiCrTi is not recommended for implant superstructures because of risk of Ni ion release to the body, and fluorides should be avoided in acidic media because Ti, Ti6Al4V, and CoCr-c superstructures show galvanic corrosion. The best combinations are Ti/Ti6Al4V and Ti/CoCr as alternative of noble gold alloys. PMID:29361767

  13. A family of cell-adhering peptides homologous to fibrinogen C-termini

    International Nuclear Information System (INIS)

    Levy-Beladev, Liron; Levdansky, Lilia; Gaberman, Elena; Friedler, Assaf; Gorodetsky, Raphael

    2010-01-01

    Research highlights: → Cell-adhesive sequences homologous to fibrinogen C-termini exist in other proteins. → The extended homologous cell-adhesive C-termini peptides family is termed Haptides. → In membrane-like environment random coiled Haptides adopt a helical conformation. → Replacing positively charged residues with alanine reduces Haptides activity. -- Abstract: A family of cell-adhesive peptides homologous to sequences on different chains of fibrinogen was investigated. These homologous peptides, termed Haptides, include the peptides Cβ, preCγ, and CαE, corresponding to sequences on the C-termini of fibrinogen chains β, γ, and αE, respectively. Haptides do not affect cell survival and rate of proliferation of the normal cell types tested. The use of new sensitive assays of cell adhesion clearly demonstrated the ability of Haptides, bound to inert matrices, to mediate attachment of different matrix-dependent cell types including normal fibroblasts, endothelial, and smooth muscle cells. Here we present new active Haptides bearing homologous sequences derived from the C-termini of other proteins, such as angiopoietin 1 and 2, tenascins C and X, and microfibril-associated glycoprotein-4. The cell adhesion properties of all the Haptides were found to be associated mainly with their 11 N-terminal residues. Mutated preCγ peptides revealed that positively charged residues account for their attachment effect. These results suggest a mechanism of direct electrostatic interaction of Haptides with the cell membrane. The extended Haptides family may be applied in modulating adhesion of cells to scaffolds for tissue regeneration and for enhancement of nanoparticulate transfection into cells.

  14. Evaluation of pH, buffering capacity, viscosity and flow rate levels of saliva in caries-free, minimal caries and nursing caries children: An in vivo study

    Directory of Open Access Journals (Sweden)

    Dwitha Animireddy

    2014-01-01

    Full Text Available Background and Aim: The present study was undertaken to evaluate the pH, buffering capacity, viscosity and flow rate of saliva in caries free, minimal caries and nursing caries children and to evaluate the relationship of these on the caries activity of children. Materials and Methods: A total of 75 school children of age group between 4 and 12 years were selected and divided into three equal groups: Group I, Group II and Group III, consisting of 25 subjects each. Group I included caries-free subjects, Group II included subjects with minimal caries and Group III included subjects with nursing caries. Saliva samples were collected from all subjects and were estimated for flow rate, pH, buffering capacity and viscosity. Results: There was a significant decrease in the mean salivary flow rate, salivary ph and salivary buffer capacity and a significant increase in the salivary viscosity among caries-free subjects, subjects with minimal caries and subjects with nursing caries. Conclusion: The physicochemical properties of saliva, such as salivary flow rate, pH, buffering capacity and viscosity, has a relation with caries activity in children and act as markers of caries activity.

  15. A short synthetic peptide fragment of human C2ORF40 has therapeutic potential in breast cancer

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Chaoyang [Shandong Univ., Jinan (China); Zhang, Pengju [Shandong Univ., Jinan (China); Jiang, Anli [Shandong Univ., Jinan (China); Mao, Jian-Hua [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Wei, Guangwei [Shandong Univ. School of Medicine, Jinan (China)

    2017-03-30

    C2ORF40 encodes a secreted protein which is cleaved to generate soluble peptides by proteolytic processing and this process is believed to be necessary for C2ORF40 to exert cell type specific biological activity. Here, we reported a short mimic peptide of human C2ORF40 acts potential therapeutic efficacy in human cancer cells in vitro and in vivo. We synthesized a short peptide of human C2ORF40, named C2ORF40 mimic peptide fragment and assessed its biological function on cancer cell growth, migration and tumorigenesis. Cell growth assay showed that C2ORF40 mimic peptide fragment significantly suppressed cell proliferation of breast and lung cancer cells. Moreover, C2ORF40 mimic peptide fragment significantly inhibited the migration and invasion of breast cancer cells. Furthermore, we showed that this peptide suppressed tumorigenesis in breast tumor xenograft model. Cell cycle assay indicated that the C2ORF40 mimic peptide fragment suppressed the growth of tumor cells through inducing mitotic phase arrest. In conclusion, our results firstly suggested that this short synthetic peptide of human C2ORF40 may be a candidate tumor therapeutic agent.

  16. Erosive potential of saliva stimulating tablets with and without fluoride in irradiated head and neck cancer patients

    DEFF Research Database (Denmark)

    Lajer, Christel; Buchwald, Christian; Nauntofte, Birgitte

    2009-01-01

    stimulated saliva without fluoride, and with fluoride. Saliva pH was determined without loss of CO(2) and in combination with inorganic measures used to calculate the degree of saturation of hydroxyapatite (HAp) and fluorapatite (FAp). EP was determined directly in all saliva samples by monitored dissolution......BACKGROUND: Patients irradiated in the head and neck region often suffer from severe dry mouth and use acidic saliva stimulating products, which may cause erosion of teeth. PURPOSE: To determine saliva stimulating effects and erosive potential (EP) of acidic saliva stimulating tablets (Xerodent...

  17. ARSENIC SPECIATION ANALYSIS IN HUMAN SALIVA

    Science.gov (United States)

    Background: Determination of arsenic species in human saliva is potentially useful for biomonitoring of human exposure to arsenic and for studying arsenic metabolism. However, there is no report on the speciation analysis of arsenic in saliva. Methods: Arsenic species in saliva ...

  18. The Association of Fasting Glucose, Insulin, and C-Peptide, with 19-Year Incidence of Coronary Heart Disease in Older Japanese-American Men; the Honolulu Heart Program

    Directory of Open Access Journals (Sweden)

    Nazneem Wahab

    2018-04-01

    Full Text Available The role of fasting glucose, insulin levels, and C-peptide in coronary heart disease (CHD in non-diabetic individuals remains uncertain. We examined the association between fasting glucose, insulin and C-peptide with the long-term incidence of CHD in Japanese-American men. In 1980–1982, from a random sample of the Honolulu Heart Program men (n = 1378, aged 61–81 years, data on several CHD and metabolic risk factors were obtained to examine the relation of fasting glucose, insulin and C-peptide to 19-year CHD incidence. Age-adjusted incidence of CHD increased with increasing quintiles of glucose, insulin and C-peptide. Age-adjusted CHD rates in the glucose quintiles were 11.9, 11.6, 14.4, 18.1 and 24.1 per 1000 person-years (trend p < 0.001. In individual Cox models (lowest quintiles of glucose, insulin and C-peptide as reference the relative risks (95% confidence interval of CHD incidence for the glucose quintiles adjusting for age, smoking, hypertension, cholesterol, physical activity, and body mass index, were 0.9 (0.6–1.4, 1.2 (0.8–1.8, 1.4 (0.9–2.2, and 1.7 (1.1–2.6, respectively (trend p = 0.004. Insulin and C-peptide were not significantly associated with CHD on multivariate analysis. Fasting glucose remained the only significant predictor of increased CHD risk (p = 0.003 in a model combining all 3 metabolic variables. In this cohort, only fasting glucose independently predicts long-term incidence of CHD. Age-adjusted insulin and C-peptide levels were associated with CHD incidence, but after adjustment for other risk factors, do not independently predict CHD.

  19. Lutzomyia longipalpis saliva or salivary protein LJM19 protects against Leishmania braziliensis and the saliva of its vector, Lutzomyia intermedia.

    Directory of Open Access Journals (Sweden)

    Natalia M Tavares

    Full Text Available BACKGROUND: Leishmania transmission occurs in the presence of insect saliva. Immunity to Phlebotomus papatasi or Lutzomyia longipalpis saliva or salivary components confers protection against an infection by Leishmania in the presence of the homologous saliva. However, immunization with Lutzomyia intermedia saliva did not protect mice against Leishmania braziliensis plus Lu. intermedia saliva. In the present study, we have studied whether the immunization with Lu. longipalpis saliva or a DNA plasmid coding for LJM19 salivary protein would be protective against L. braziliensis infection in the presence of Lu. intermedia saliva, the natural vector for L. braziliensis. METHODOLOGY/PRINCIPAL FINDINGS: Immunization with Lu. longipalpis saliva or with LJM19 DNA plasmid induced a Delayed-Type Hypersensitivity (DTH response against Lu. longipalpis as well as against a Lu. intermedia saliva challenge. Immunized and unimmunized control hamsters were then intradermally infected in the ears with L. braziliensis in the presence of Lu. longipalpis or Lu. intermedia saliva. Animals immunized with Lu. longipalpis saliva exhibited smaller lesion sizes as well as reduced disease burdens both at lesion site and in the draining lymph nodes. These alterations were associated with a significant decrease in the expression levels of IL-10 and TGF-β. Animals immunized with LJM19 DNA plasmid presented similar findings in protection and immune response and additionally increased IFN-γ expression. CONCLUSIONS/SIGNIFICANCE: Immunization with Lu. longipalpis saliva or with a DNA plasmid coding LJM19 salivary protein induced protection in hamsters challenged with L. braziliensis plus Lu. intermedia saliva. These findings point out an important role of immune response against saliva components, suggesting the possibility to develop a vaccine using a single component of Lu. longipalpis saliva to generate protection against different species of Leishmania, even those

  20. Incidence of Epstein-Barr Virus in Astronaut Saliva During Spaceflight

    Science.gov (United States)

    Payne, Deborah A.; Mehta, Satish K.; Tyring, Stephen K.; Stowe, Raymond P.; Pierson, Duane L.

    1998-01-01

    Astronauts experience psychological and physical stresses that may result in re-activation of latent viruses during spaceflight, potentially increasing the risk of disease among crew members. The shedding of Epstein-Barr virus (EBV) in the saliva of astronauts will increase during spaceflight. A total of 534 saliva specimens were collected from 11 EBV-seropositive astronauts before, during, and after four space shuttle missions. The presence of EBV DNA in saliva, assessed by polymerase chain reaction (PCR), was used to determine shedding patterns before, during, and after spaceflight. EBV DNA was detected more frequently before flight than during (p less than 0.001) or after (p less than 0.01) flight. No significant difference between the in-flight and postflight periods was detected in the frequency of occurrence of EBV DNA. The increased frequency of shedding of EBV before flight suggests that stress levels may be greater before launch than during or after spaceflight.

  1. Metabolism and pharmacokinetic of cyclo-peptides and peptides. Use of radioelement and stable isotopes

    International Nuclear Information System (INIS)

    Aninat, C.

    2003-10-01

    More and more peptides and proteins are used in therapeutic. Three mainly techniques are used for pharmacokinetic and metabolism studies: immunoassay, radioactively labeled molecules and mass spectrometry. In the first part of this work, we have used uniformly labelled peptides (C-peptide and insulin) with stables ( 13 C, 15 N, and 13 C/ 15 N) or radioactive ( 14 C) isotopes to investigated these kind of studies. These works are based on isotope dilution mass spectrometry assay. In a second time we have investigated the metabolism of a particular cyclo-peptides families composed of two amino acids: the diketo-piperazine. These compounds are found in mammals and in microorganisms. There are not recognized by proteolytic enzymes. We have estimated if the main enzymes implicated in the metabolism of xenobiotics, the P450 cytochrome mono-oxygenases, were able to recognized them

  2. Protein buffering in model systems and in whole human saliva.

    Directory of Open Access Journals (Sweden)

    Andreas Lamanda

    Full Text Available The aim of this study was to quantify the buffer attributes (value, power, range and optimum of two model systems for whole human resting saliva, the purified proteins from whole human resting saliva and single proteins. Two model systems, the first containing amyloglucosidase and lysozyme, and the second containing amyloglucosidase and alpha-amylase, were shown to provide, in combination with hydrogencarbonate and di-hydrogenphosphate, almost identical buffer attributes as whole human resting saliva. It was further demonstrated that changes in the protein concentration as small as 0.1% may change the buffer value of a buffer solution up to 15 times. Additionally, it was shown that there was a protein concentration change in the same range (0.16% between saliva samples collected at the time periods of 13:00 and others collected at 9:00 am and 17:00. The mode of the protein expression changed between these samples corresponded to the change in basic buffer power and the change of the buffer value at pH 6.7. Finally, SDS Page and Ruthenium II tris (bathophenantroline disulfonate staining unveiled a constant protein expression in all samples except for one 50 kDa protein band. As the change in the expression pattern of that 50 kDa protein band corresponded to the change in basic buffer power and the buffer value at pH 6.7, it was reasonable to conclude that this 50 kDa protein band may contain the protein(s belonging to the protein buffer system of human saliva.

  3. Characterization of an insulin-like growth factor-I/somatomedin-C radioimmunoassay specific for the C-peptide region

    International Nuclear Information System (INIS)

    Hintz, R.L.; Liu, F.; Seegan, G.

    1982-01-01

    Insulin-like growth factor-I (IGF-I) and somatomedin-C (SM-C) have been shown to be functionally identical by a number of criteria. We have synthesized the 12 amino acid C-peptide region of IGF-I (Gly-Tyr-Gly-Ser-Ser-Ser-Arg-Arg-Ala-Pro-Glu-Thr) and developed a RIA based on antibodies against this synthetic peptide. IGF-I and SM-C were indistinguishable in this RIA. No other peptides competed for this antiserum. The SM-C/IGF-I values of acid-chromatographed serum were strongly age dependent. The mean of children 1-5 yr old was 0.67 +/- 0.033 U/ml (mean +/- sD; n = 23), whereas the mean of children 12-17 yr old was 2.01 +/- 0.66 U/ml (n = 39) and the mean of 38 adults 26-85 yr old was 1.05 +/- 0.34. The SM-C/IGF-I values measured by this RIA were also growth hormone dependent. Thus, this region-specific RIA provides a clinically useful assessment of serum SM-C/IGF-I levels

  4. Selected antibacterial factors in the saliva of diabetic patients.

    Science.gov (United States)

    Malicka, Barbara; Kaczmarek, Urszula; Skośkiewicz-Malinowska, Katarzyna

    2015-03-01

    Diabetes mellitus leads to many systemic complications, including changes in the morphology, function of the salivary glands, and the composition of saliva. The study comprised a randomly selected 156 adults, of both genders, aged from 21 to 79, out of which patients with diabetes type 1 and 2, and healthy subjects forming two control age- and gender matched to the ill subjects. In unstimulated mixed saliva, total protein, peroxidase, myeloperoxidase and immunoglobulin A were measured as well as salivary flow rate. The periodontal condition was assessed with the use of GI, mSBI and PSR index. The obtained data were analysed with the use of U Mann-Whitney's test, Spearman's rang correlation and Chi-square test at a significant level of p diabetics in comparison to healthy age and gender matched control group had a lower salivary flow rate (p diabetics in comparison to control subjects had a higher level of total protein concentration (p diabetes type 1 and 2 can cause abnormalities in salivary glands function resulting in the diminishing of salivary flow rate and the increase in total protein content. Higher levels of myeloperoxidase and IgA in the saliva can be linked to worse periodontal condition in the diabetic patients. Copyright © 2014. Published by Elsevier Ltd.

  5. Shear bond strength of metallic brackets: influence of saliva contamination

    Directory of Open Access Journals (Sweden)

    Luciana Borges Retamoso

    2009-06-01

    Full Text Available OBJECTIVE: To evaluate the influence of saliva contamination on shear bond strength and the bond failure pattern of 3 adhesive systems (Transbond XT, AdheSE and Xeno III on orthodontic metallic brackets bonded to human enamel. MATERIAL AND METHODS: Seventy-two permanent human molars were cut longitudinally in a mesiodistal direction, producing seventy-two specimens randomly divided into six groups. Each system was tested under 2 different enamel conditions: no contamination and contaminated with saliva. In T, A and X groups, the adhesive systems were applied to the enamel surface in accordance with manufacturer's instructions. In TS, AS and XS groups, saliva was applied to enamel surface followed by adhesive system application. The samples were stored in distilled water at 37ºC for 24 h, and then tested for shear bond strength in a universal testing machine (Emic, DL 2000 running at a crosshead speed of 1 mm/min. After bond failure, the enamel surfaces were observed under an optical microscope at 40x magnification. RESULTS: The control and contaminated groups showed no significant difference in shear bond strength for the same adhesive system. However, shear bond strength of T group (17.03±4.91 was significantly higher than that of AS (8.58±1.73 and XS (10.39±4.06 groups (p<0.05. Regarding the bond failure pattern, TS group had significantly higher scores of no adhesive remaining on the tooth in the bonding area than other groups considering the adhesive remnant index (ARI used to evaluate the amount of adhesive left on the enamel. CONCLUSIONS: Saliva contamination showed little influence on the 24-h shear bond strength of orthodontic brackets.

  6. Quantification of anti-Leishmania antibodies in saliva of dogs.

    Science.gov (United States)

    Cantos-Barreda, Ana; Escribano, Damián; Bernal, Luis J; Cerón, José J; Martínez-Subiela, Silvia

    2017-08-15

    Detection of serum anti-Leishmania antibodies by quantitative or qualitative techniques has been the most used method to diagnose Canine Leishmaniosis (CanL). Nevertheless, saliva may represent an alternative to blood because it is easy to collect, painless and non-invasive in comparison with serum. In this study, two time-resolved immunofluorometric assays (TR-IFMAs) for quantification of anti-Leishmania IgG2 and IgA antibodies in saliva were developed and validated and their ability to distinguish Leishmania-seronegative from seropositive dogs was evaluated. The analytical study was performed by evaluation of assay precision, sensitivity and accuracy. In addition, serum from 48 dogs (21 Leishmania-seropositive and 27 Leishmania-seronegative) were analyzed by TR-IFMAs. The assays were precise, with an intra- and inter-assay coefficients of variation lower than 11%, and showed high level of accuracy, as determined by linearity under dilution (R 2 =0.99) and recovery tests (>88.60%). Anti-Leishmania IgG2 antibodies in saliva were significantly higher in the seropositive group compared with the seronegative (pLeishmania IgA antibodies between both groups were observed. Furthermore, TR-IFMA for quantification of anti-Leishmania IgG2 antibodies in saliva showed higher differences between seropositive and seronegative dogs than the commercial assay used in serum. In conclusion, TR-IFMAs developed may be used to quantify anti-Leishmania IgG2 and IgA antibodies in canine saliva with an adequate precision, analytical sensitivity and accuracy. Quantification of anti-Leishmania IgG2 antibodies in saliva could be potentially used to evaluate the humoral response in CanL. However, IgA in saliva seemed not to have diagnostic value for this disease. For future studies, it would be desirable to evaluate the ability of the IgG2 assay to detect dogs with subclinical disease or with low antibody titers in serum and also to study the antibodies behaviour in saliva during the

  7. Characterization of multiple antilisterial peptides produced by sakacin P-producing Lactobacillus sakei subsp. sakei 2a.

    Science.gov (United States)

    Carvalho, Kátia G; Bambirra, Felipe H S; Nicoli, Jacques R; Oliveira, Jamil S; Santos, Alexandre M C; Bemquerer, Marcelo P; Miranda, Antonio; Franco, Bernadette D G M

    2018-05-01

    Antimicrobial compounds produced by lactic acid bacteria can be explored as natural food biopreservatives. In a previous report, the main antimicrobial compounds produced by the Brazilian meat isolate Lactobacillus sakei subsp. sakei 2a, i.e., bacteriocin sakacin P and two ribosomal peptides (P2 and P3) active against Listeria monocytogenes, were described. In this study, we report the spectrum of activity, molecular mass, structural identity and mechanism of action of additional six antilisterial peptides produced by Lb. sakei 2a, detected in a 24 h-culture in MRS broth submitted to acid treatment (pH 1.5) and proper fractionation and purification steps for obtention of free and cell-bound proteins. The six peptides presented similarity to different ribosomal proteins of Lb. sakei subsp sakei 23K and the molecular masses varied from 4.6 to 11.0 kDa. All peptides were capable to increase the efflux of ATP and decrease the membrane potential in Listeria monocytogenes. The activity of a pool of the obtained antilisterial compounds [enriched active fraction (EAF)] against Listeria monocytogenes in a food model (meat gravy) during refrigerated storage (4 °C) for 10 days was also tested and results indicated that the populations of L. monocytogenes in the food model containing the acid extract remained lower than those at time 0-day, evidencing that the acid extract of a culture of Lb. sakei 2a is a good technological alternative for the control of growth of L. monocytogenes in foods.

  8. Corticosteroids in plasma and saliva : the influence of oral contraceptive use and pregnancy

    NARCIS (Netherlands)

    Meulenberg, E.P.M.M.

    1995-01-01

    <p>Cortisol in saliva is considered to be derived from the free, unbound fraction in plasma by simple diffusion through the salivary gland. Despite considerable conversion into cortisone in the salivary gland by the enzyme 11β-hydroxysteroid dehydrogenase, levels of cortisol in saliva form a

  9. Hubungan PH, Hidrasi, Kapasitas Bufer Saliva, Jumlah Streptococcus Mutans dengan Keparahan Karies pada Anak Rampan Karies

    OpenAIRE

    Subekti, Ani; Kristiani, Nany; Rimbyastuti, Hermin

    2013-01-01

    Rampan caries is the habit of bottle feeding at night, sweets preference and often meals sucking. Saliva is one of the factors that may influence the occurrence of caries. Some of the factors that cause caries saliva such as saliva pH, hydration, microorganisms of the oral cavity and salivary buffer capacity.The research objective was to determine the relationship of hydration, pH, buffer capacity salivary and growth Streptoccoccus mutans against caries severity or deft.A sample of 30 kinderg...

  10. Structure of a C-terminal AHNAK peptide in a 1:2:2 complex with S100A10 and an acetylated N-terminal peptide of annexin A2

    International Nuclear Information System (INIS)

    Ozorowski, Gabriel; Milton, Saskia; Luecke, Hartmut

    2013-01-01

    Structure of a 20-amino-acid peptide of AHNAK bound asymmetrically to the AnxA2–S100A10A heterotetramer (1:2:2 symmetry) provides insights into the atomic level interactions that govern this membrane-repair scaffolding complex. AHNAK, a large 629 kDa protein, has been implicated in membrane repair, and the annexin A2–S100A10 heterotetramer [(p11) 2 (AnxA2) 2 )] has high affinity for several regions of its 1002-amino-acid C-terminal domain. (p11) 2 (AnxA2) 2 is often localized near the plasma membrane, and this C2-symmetric platform is proposed to be involved in the bridging of membrane vesicles and trafficking of proteins to the plasma membrane. All three proteins co-localize at the intracellular face of the plasma membrane in a Ca 2+ -dependent manner. The binding of AHNAK to (p11) 2 (AnxA2) 2 has been studied previously, and a minimal binding motif has been mapped to a 20-amino-acid peptide corresponding to residues 5654–5673 of the AHNAK C-terminal domain. Here, the 2.5 Å resolution crystal structure of this 20-amino-acid peptide of AHNAK bound to the AnxA2–S100A10 heterotetramer (1:2:2 symmetry) is presented, which confirms the asymmetric arrangement first described by Rezvanpour and coworkers and explains why the binding motif has high affinity for (p11) 2 (AnxA2) 2 . Binding of AHNAK to the surface of (p11) 2 (AnxA2) 2 is governed by several hydrophobic interactions between side chains of AHNAK and pockets on S100A10. The pockets are large enough to accommodate a variety of hydrophobic side chains, allowing the consensus sequence to be more general. Additionally, the various hydrogen bonds formed between the AHNAK peptide and (p11) 2 (AnxA2) 2 most often involve backbone atoms of AHNAK; as a result, the side chains, particularly those that point away from S100A10/AnxA2 towards the solvent, are largely interchangeable. While the structure-based consensus sequence allows interactions with various stretches of the AHNAK C-terminal domain, comparison

  11. Sequence of the radioactive tryptic peptide obtained after inactivating the F1-ATPase of the thermophilic bacterium PS3 with 5'-p-fluorosulfonylbenzoyl[3H]adenosine at 65 degrees C

    International Nuclear Information System (INIS)

    Bullough, D.A.; Yoshida, M.; Allison, W.S.

    1986-01-01

    Following a lag of about 30 min, the F1-ATPase from the thermophilic bacterium, PS3 (TF1), was inactivated slowly by 0.8 mM 5'-p-fluorosulfonylbenzoyladenosine (FSBA) at 23 degrees C and pH 7.0. When the enzyme was treated with 0.2 mM FSBA at pH 7.0 and 23 degrees C for 15 min and gel-filtered, no enzyme activity was lost. However, the lag in inactivation was abolished when the enzyme was subsequently incubated with 2.0 mM FSBA at 23 degrees C in the pH range from 6.8 to 10.0. The pH-inactivation profile obtained under these conditions revealed a pK alpha of about 9.3 which was associated with the inactivation. When pretreated TF1 was inactivated at 23 degrees C with [3H]FSBA by about 90%, greater than 20 mol of [3H]SBA was incorporated per mole of enzyme. TF1 was inactivated rapidly by 0.8 mM FSBA at pH 6.4 and 65 degrees C, and no lag was observed. Following inactivation of TF1 with 0.8 mM [3H]FSBA at 65 degrees C and pH 6.4, about 10 mol of [3H]SBA was incorporated per mole of enzyme. When a tryptic digest of the labeled enzyme was fractionated by reversed-phase high-performance liquid chromatography, a single major radioactive peptide was isolated. When subjected to automatic Edman degradation, this peptide was shown to have the amino acid sequence: A-L-A-P-E-I-V-G-E-E-H-X-Q-V-A-R, where X indicates that a phenylthiohydantoin derivative was not detected in cycle 12. However, from the DNA sequence of the gene encoding the subunit of TF1 (Y. Kagawa, M. Ishizuka, T. Saishu, and S. Nakao (1985)), this position has been shown to be occupied by tyrosine. This tyrosine is homologous with beta-Tyr-368 of the bovine mitochondrial F1-ATPase (MF1) the modification of which is responsible for the inactivation MF1 by FSBA

  12. Rapid antemortem detection of CWD prions in deer saliva.

    Directory of Open Access Journals (Sweden)

    Davin M Henderson

    Full Text Available Chronic wasting disease (CWD is an efficiently transmitted prion disease of cervids, now identified in 22 United States, 2 Canadian provinces and Korea. One hallmark of CWD is the shedding of infectious prions in saliva, as demonstrated by bioassay in deer. It is also clear that the concentration of prions in saliva, blood, urine and feces is much lower than in the nervous system or lymphoid tissues. Rapid in vitro detection of CWD (and other prions in body fluids and excreta has been problematic due to the sensitivity limits of direct assays (western blotting, ELISA and the presence of inhibitors in these complex biological materials that hamper detection. Here we use real-time quaking induced conversion (RT-QuIC to demonstrate CWD prions in both diluted and prion-enriched saliva samples from asymptomatic and symptomatic white-tailed deer. CWD prions were detected in 14 of 24 (58.3% diluted saliva samples from CWD-exposed white-tailed deer, including 9 of 14 asymptomatic animals (64.2%. In addition, a phosphotungstic acid enrichment enhanced the RT-QuIC assay sensitivity, enabling detection in 19 of 24 (79.1% of the above saliva samples. Bioassay in Tg[CerPrP] mice confirmed the presence of infectious prions in 2 of 2 RT-QuIC-positive saliva samples so examined. The modified RT-QuIC analysis described represents a non-invasive, rapid ante-mortem detection of prions in complex biologic fluids, excreta, or environmental samples as well as a tool for exploring prion trafficking, peripheralization, and dissemination.

  13. pMPES: A Modular Peptide Expression System for the Delivery of Antimicrobial Peptides to the Site of Gastrointestinal Infections Using Probiotics

    Directory of Open Access Journals (Sweden)

    Kathryn Geldart

    2016-10-01

    Full Text Available Antimicrobial peptides are a promising alternative to traditional antibiotics, but their utility is limited by high production costs and poor bioavailability profiles. Bacterial production and delivery of antimicrobial peptides (AMPs directly at the site of infection may offer a path for effective therapeutic application. In this study, we have developed a vector that can be used for the production and secretion of seven antimicrobial peptides from both Escherichia coli MC1061 F’ and probiotic E.coli Nissle 1917. The vector pMPES (Modular Peptide Expression System employs the Microcin V (MccV secretion system and a powerful synthetic promoter to drive AMP production. Herein, we demonstrate the capacity of pMPES to produce inhibitory levels of MccV, Microcin L (MccL, Microcin N (McnN, Enterocin A (EntA, Enterocin P (EntP, Hiracin JM79 (HirJM79 and Enterocin B (EntB. To our knowledge, this is the first demonstration of such a broadly-applicable secretion system for AMP production. This type of modular expression system could expedite the development of sorely needed antimicrobial technologies

  14. Fall in C-Peptide During First 4 Years From Diagnosis of Type 1 Diabetes: Variable Relation to Age, HbA1c, and Insulin Dose.

    Science.gov (United States)

    Hao, Wei; Gitelman, Steven; DiMeglio, Linda A; Boulware, David; Greenbaum, Carla J

    2016-10-01

    We aimed to describe the natural history of residual insulin secretion in Type 1 Diabetes TrialNet participants over 4 years from diagnosis and relate this to previously reported alternative clinical measures reflecting β-cell secretory function. Data from 407 subjects from 5 TrialNet intervention studies were analyzed. All subjects had baseline stimulated C-peptide values of ≥0.2 nmol/L from mixed-meal tolerance tests (MMTTs). During semiannual visits, C-peptide values from MMTTs, HbA1c, and insulin doses were obtained. The percentage of individuals with stimulated C-peptide of ≥0.2 nmol/L or detectable C-peptide of ≥0.017 nmol/L continued to diminish over 4 years; this was markedly influenced by age. At 4 years, only 5% maintained their baseline C-peptide secretion. The expected inverse relationships between C-peptide and HbA1c or insulin doses varied over time and with age. Combined clinical variables, such as insulin-dose adjusted HbA1c (IDAA1C) and the relationship of IDAA1C to C-peptide, also were influenced by age and time from diagnosis. Models using these clinical measures did not fully predict C-peptide responses. IDAA1C ≤9 underestimated the number of individuals with stimulated C-peptide ≥0.2 nmol/L, especially in children. Current trials of disease-modifying therapy for type 1 diabetes should continue to use C-peptide as a primary end point of β-cell secretory function. Longer duration of follow-up is likely to provide stronger evidence of the effect of disease-modifying therapy on preservation of β-cell function. © 2016 by the American Diabetes Association.

  15. cDNA, deduced polypeptide structure and chromosomal assignment of human pulmonary surfactant proteolipid, SPL(pVal)

    International Nuclear Information System (INIS)

    Glasser, S.W.; Korfhagen, T.R.; Weaver, T.E.; Clark, J.C.; Pilot-Matias, T.; Meuth, J.; Fox, J.L.; Whitsett, J.A.

    1988-01-01

    In hyaline membrane disease of premature infants, lack of surfactant leads to pulmonary atelectasis and respiratory distress. Hydrophobic surfactant proteins of M/sub r/ = 5000-14,000 have been isolated from mammalian surfactants which enhance the rate of spreading and the surface tension lowering properties of phospholipids during dynamic compression. The authors have characterized the amino-terminal amino acid sequence of pulmonary proteolipids from ether/ethanol extracts of bovine, canine, and human surfactant. Two distinct peptides were identified and termed SPL(pVal) and SPL(Phe). An oligonucleotide probe based on the valine-rich amino-terminal amino acid sequence of SPL(pVal) was utilized to isolate cDNA and genomic DNA encoding the human protein, termed surfactant proteolipid SPL(pVal) on the basis of its unique polyvaline domain. The primary structure of a precursor protein of 20,870 daltons, containing the SPL(pVal) peptide, was deduced from the nucleotide sequence of the cDNAs. Hybrid-arrested translation and immunoprecipitation of labeled translation products of human mRNA demonstrated a precursor protein, the active hydrophobic peptide being produced by proteolytic processing. Two classes of cDNAs encoding SPL(pVal) were identified. Human SPL(pVal) mRNA was more abundant in the adult than in fetal lung. The SPL(pVal) gene locus was assigned to chromosome 8

  16. [Activity of alpha-amylase and concentration of protein in saliva of pregnant women].

    Science.gov (United States)

    Ciejak, Magdalena; Olszewska, Maria; Jakubowska, Katarzyna; Zebiełowicz, Dariusz; Safranow, Krzysztof; Chlubek, Dariusz

    2007-01-01

    One of the hypothetical reasons of the increased incidence of caries in women during the pregnancy may be the increased activity of alpha-amylase, which can be found in their saliva. The enzyme takes part in the process of decomposition of simple sugars, which make basic substrate for caries-causing bacteria. The aim of the paper was the evaluation of the influence of pregnancy and gestational age on the activity of alpha-amylase and the concentration of protein in women's saliva. The examined group consisted of 64 pregnant women at age 17-39, between 21st and 40th week of pregnancy. The control group consisted of 44 healthy women at age 20-35, who were not pregnant. In saliva, which was taken before morning meal, without stimulation, protein concentration was determined by Bradford method and the activity of amylase was determined by kinetic method. The activity of amylase correlated strongly and positively with protein concentration in saliva of both the pregnant (RS = +0.65; p women. There were no significant differences between examined parameters in the examined and the control group. It has been observed in the examined group, that there is the significant negative correlation between protein concentration in saliva and the week of pregnancy (RS = -0.35; p increased caries incidence of pregnant women. However, the observed changes of total protein concentration in saliva during pregnancy, suggest that the exact cognition of proteins in pregnant women's saliva may reveal new mechanisms, which lead to an increase of caries risk.

  17. Pseudomonas aeruginosa elastase cleaves a C-terminal peptide from human thrombin that inhibits host inflammatory responses

    DEFF Research Database (Denmark)

    van der Plas, Mariena J A; Bhongir, Ravi K V; Kjellström, Sven

    2016-01-01

    Pseudomonas aeruginosa is an opportunistic pathogen known for its immune evasive abilities amongst others by degradation of a large variety of host proteins. Here we show that digestion of thrombin by P. aeruginosa elastase leads to the release of the C-terminal thrombin-derived peptide FYT21...

  18. Streptococcal adhesin SspA/B analogue peptide inhibits adherence and impacts biofilm formation of Streptococcus mutans.

    Directory of Open Access Journals (Sweden)

    Tatsuro Ito

    Full Text Available Streptococcus mutans, the major causative agent of dental caries, adheres to tooth surfaces via the host salivary glycoprotein-340 (gp340. This adherence can be competitively inhibited by peptides derived from the SspA/B adhesins of Streptococcus gordonii, a human commensal microbe that competes for the same binding sites. Ssp(A4K-A11K, a double-lysine substituted SspA/B peptide analogue, has been shown to exhibit superior in vitro binding affinity for a gp340-derived peptide (SRCRP2, suggesting that Ssp(A4K-A11K may be of clinical interest. In the present work, we tested the inhibitory effects of Ssp(A4K-A11K on adherence and biofilm formation of S. mutans by reconstructing an artificial oral environment using saliva-coated polystyrene plates and hydroxyapatite disks. Bacterial adherence (adherence period: 1 h was assessed by an enzyme-linked immunosorbent assay using biotinylated bacterial cells. Biofilm formation (periods: 8, 11, or 14 h was assessed by staining and imaging of the sessile cells, or by recovering biofilm cells and plating for cell counts. The pH values of the culture media were measured as a biofilm acidogenicity indicator. Bactericidality was measured by loss of optical density during culturing in the presence of the peptide. We observed that 650 μM Ssp(A4K-A11K significantly inhibited adherence of S. mutans to saliva-coated polystyrene; a similar effect was seen on bacterial affinity for SRCRP2. Ssp(A4K-A11K had lesser effects on the adherence of commensal streptococci. Pretreatment of polystyrene and hydroxyapatite with 650 μM Ssp(A4K-A11K significantly attenuated biofilm formation, whether tested with glucose- or sucrose-containing media. The SspA/B peptide's activity did not reflect bactericidality. Strikingly, pH in Ssp-treated 8-h (6.8 ± 0.06 and 11-h (5.5 ± 0.06 biofilms showed higher values than the critical pH. Thus, Ssp(A4K-A11K acts by inhibiting bacterial adherence and cariogrnic biofilm formation. We further

  19. Effectiveness of transcutaneous electrical nerve stimulation on saliva production in post-radiated oral cancer patients

    Directory of Open Access Journals (Sweden)

    Sakshi Ojha

    2016-01-01

    Full Text Available Aims and Objectives: To determine the effectiveness of transcutaneous electrical nerve stimulation (TENS in stimulating salivary flow in post-radiated oral cancer patients, and to compare the salivary flow rate between unstimulated saliva and saliva stimulated with TENS in post-radiated oral cancer patients. Materials and Methods: In 30 patients who underwent radiotherapy for oral cancer, unstimulated saliva was collected every minute for 5 min in a graduated test tube. The TENS unit was activated and stimulated saliva was collected for 5 min in a separate graduated test tube, and the flow rate was compared with the unstimulated salivary flow rate. Results: A statistically significant improvement was seen in saliva production during stimulation (P < 0.001. In addition, statistically significant increase in TENS stimulated saliva was observed in patients aged ≥50 years compared to that in patients aged <50 years (P < 0.05. There was no significant difference in salivary flow rate between the two genders in both stimulated and unstimulated conditions, however, statistically significant increase in salivary flow rate was observed in males under stimulated condition (P < 0.01. Conclusion: TENS was highly effective in stimulating the whole salivary flow rate in post-radiated oral cancer patients. It is an effective supportive treatment modality in xerostomia patients caused by radiotherapy in oral cancer patients.

  20. Effect of dietary roughage level on chewing activity, ruminal pH, and saliva secretion in lactating Holstein cows.

    Science.gov (United States)

    Jiang, F G; Lin, X Y; Yan, Z G; Hu, Z Y; Liu, G M; Sun, Y D; Liu, X W; Wang, Z H

    2017-04-01

    Increasing dietary roughage level is a commonly used strategy to prevent subacute ruminal acidosis. We hypothesized that high-roughage diets could promote chewing activity, saliva secretion, and hence more alkaline to buffer rumen pH. To verify the hypothesis, 12 multiparous Holstein cows in mid lactation were randomly allocated to 4 treatments in a triplicated 4 × 4 Latin square experiment with one cow in each treatment surgically fitted with a ruminal cannula. Treatments were diets containing 40, 50, 60, or 70% of roughage on a DM basis. Increasing dietary roughage level decreased DM, CP, OM, starch, and NE L intake, increased ADF intake, and decreased milk yield linearly. Intake of NDF was quite stable across treatments and ranged from 7.8 to 8.1 kg/d per cow. Daily eating time increased linearly with increased roughage level. The increase in eating time was due to increased eating time per meal but not number of meals per day, which was stable and ranged from 8.3 to 8.5 meals per day across treatments. Increasing dietary roughage level had no effect on ruminating time (min/d), the number of ruminating periods (rumination periods per d), and chewing time per ruminating period (min/ruminating period). Ruminating time per kilogram of NDF intake and total chewing time per kilogram of ADF intake were similar across treatments (57.4 and 183.8 min/kg, respectively). Increasing dietary roughage level linearly increased daily total chewing time; linearly elevated the mean, maximum, and minimum ruminal pH; and linearly decreased total VFA concentration and molar proportion of propionate in ruminal fluid. Saliva secretion during eating was increased, the secretion during rumination was unaffected, but the secretion during resting tended to decrease with increased dietary roughage level. As a result, total saliva secretion was not affected by treatments. In conclusion, the results of the present study did not support the concept that high-roughage diets elevated ruminal p

  1. Circulating Dopamine and C-Peptide Levels in Fasting Nondiabetic Hypertensive Patients

    OpenAIRE

    Tomaschitz, Andreas; Ritz, Eberhard; Kienreich, Katharina; Pieske, Burkert; M?rz, Winfried; Boehm, Bernhard O.; Drechsler, Christiane; Meinitzer, Andreas; Pilz, Stefan

    2012-01-01

    OBJECTIVE Accumulating evidence supports a potential role for dopamine in the regulation of insulin secretion. We examined the association between circulating dopamine and C-peptide concentrations using data from the Graz Endocrine Causes of Hypertension (GECOH) study. RESEARCH DESIGN AND METHODS After 12 h of fasting, we measured plasma dopamine and serum C-peptide levels and established determining factors of insulin secretion in 201 nondiabetic hypertensive patients (mean age 48.1 ? 16.0 y...

  2. PERBEDAAN PH SALIVA ANTARA PENGGUNA PASTA GIGI YANG MENGANDUNG BAKING SODA DAN PENGGUNA PASTA GIGI YANG MENGANDUNG FLUOR

    OpenAIRE

    LINARDI, ALICIA NADIA

    2014-01-01

    2014 Latar belakang : Baking soda dan fluor merupakan bahan yang biasa ditambahkan dalam pasta gigi. Baking soda dan fluor mempunyai kemampuan untuk meningkatkan sekresi saliva dan pH saliva. Tujuan penelitian ini adalah untuk mengetahui perbedaan pH saliva antara pengguna pasta gigi yang mengandung baking soda dan pengguna pasta gigi yang mengandun fluor. Bahan dan metode : Jenis penelitian ini adalah eksperimental dengan desai...

  3. Creatine metabolism: detection of creatine and guanidinoacetate in saliva of healthy subjects.

    Science.gov (United States)

    Martínez, Lidia D; Bezard, Miriam; Brunotto, Mabel; Dodelson de Kremer, Raquel

    2016-04-01

    Creatine (Cr) plays an important role in storage and transmission of phosphate-bound energy. Cerebral creatine deficiency syndromes comprise three inherited defects in Cr biosynthesis and transport. The aim of this study was to investigate whether Cr and Guanidinoacetate (GAA) can be detected in saliva of healthy subjects and to establish the relationship between salivary and plasma levels of these molecules. An adapted gas chromatography (GC) method is described for the quantification of Cr and GAA biomarkers in saliva. Reference values were established for GAA and Cr in saliva. These values were age dependent (p= 0.001). No difference between genders was observed. We detected a difference between GAA and Cr concentrations in saliva and in plasma. The GC method for simultaneous determination of GAA and Cr in human saliva is fast, reliable, sensitive, non-invasive and precise to use as a biochemical approach in early detection of cerebral creatine deficiency syndromes. Sociedad Argentina de Investigación Odontológica.

  4. Candida albicans adherence to resin-composite restorative dental material: influence of whole human saliva.

    Science.gov (United States)

    Maza, José Luis; Elguezabal, Natalia; Prado, Carlota; Ellacuría, Joseba; Soler, Iñaki; Pontón, José

    2002-11-01

    Attachment of Candida albicans to oral surfaces is believed to be a critical event in the colonization of the oral cavity and in the development of oral diseases such as Candida-associated denture stomatitis. Although there is considerable information about the adhesion of C albicans to buccal epithelial cells and prosthetic materials, there is very little information about the adhesion of C albicans to composite restorative materials. The purpose of this study was to investigate the degree of adhesion of C albicans to a resin-composite restorative material (Herculite). The adhesion of 2 strains of C albicans, a germinative and a germ tube-deficient mutant, was studied by a visual method after incubating the fungus and the resin with and without human whole saliva. In absence of saliva, the adhesion of the C albicans germinative isolate to the resin showed an increase in parallel with the germination, reaching a maximum at the end of the experiment (120 minutes). However, no significant differences were observed in the adhesion of the agerminative mutant during the period of time studied. In the presence of saliva, the adhesion of both isolates to the resin was significantly lowered. Germination and the presence of human whole saliva are important factors in the adhesion of C albicans to the resin-composite restorative material Herculite.

  5. Uranyl Photocleavage of Phosphopeptides Yields Truncated C-Terminally Amidated Peptide Products

    DEFF Research Database (Denmark)

    Elnegaard, Rasmus L B; Møllegaard, Niels Erik; Zhang, Qiang

    2017-01-01

    photocleavage reaction of a tetraphosphorylated β-casein model peptide. We show that the primary photocleavage products of the uranyl-catalysed reaction are C-terminally amidated. This could be of great interest to the pharmaceutical industry, as efficient peptide amidation reactions are one of the top...

  6. Saliva, Serum Levels of Interleukin-21, -33 and Prostaglandin E2 in Patients with Generalised Aggressive or Chronic Periodontitis.

    Science.gov (United States)

    Gümüş, Pınar; Nizam, Nejat; Nalbantsoy, Ayşe; Özçaka, Özgün; Buduneli, Nurcan

    This cross-sectional study aims to evaluate saliva, serum levels of interleukin-21 (IL-21), IL-33, and prostaglandin E2 (PGE2) in patients with generalised chronic periodontitis or aggressive periodontitis. Before initiation of any periodontal treatment, saliva and serum samples were collected and clinical periodontal measurements were recorded from 94 participants (25 aggressive periodontitis patients, 25 chronic periodontitis patients, 44 periodontally healthy individuals). IL-21, IL-33 and PGE2 levels in serum and saliva samples were determined by ELISA. Data were tested statistically using Kruskal-Wallis, Mann-Whitney U-, and Spearman-rho rank tests. Saliva IL-33 levels were statistically significantly higher in the chronic than the aggressive group (p periodontitis groups. Saliva IL-33 levels correlated with age in the chronic periodontitis group (p periodontitis groups (p chronic and aggressive periodontitis, but the present findings support the role of these cytokines in periodontitis. Statistically significantly higher saliva IL-33 levels in the chronic periodontitis group warrant further research.

  7. Increased [32P]-phosphorylation of tryptic peptides of erythrocyte spectrin in Duchenne muscular dystrophy

    International Nuclear Information System (INIS)

    Mabry, M.E.; Roses, A.D.

    1981-01-01

    Increased [32P]-incorporation in tryptic peptides of the erythrocyte membrane protein spectrin Band 2 in Duchenne muscular dystrophy (DMD) was studied in a consecutive series of 10 matched DMD/control pairs. Spectrin was [32P]-phosphorylated by cyclic AMP-independent endogenous membrane protein kinase in the presence of [gamma-32P]ATP. [32P]-labeled spectrin was isolated, purified, and subjected to tryptic cleavage with excess trypsin. The resulting peptides were separated on a high-resolution 5%/15% stacking SDS--polyacrylamide gel electrophoresis system. Liquid scintillation counting was performed on sequential slices of unstained gels. A broad [32P]-labeled band containing a number of [32P]-polypeptides was found to be more highly [32P]-phosphorylated in DMD patients than in their matched controls. This band migrated with an apparent molecular mass of 4.8-5.2 kilodaltons and contained approximately 55% of total [32P] radioactivity covalently bound to spectrin peptides. These data demonstrated an increased [32P]-phosphorylation of an identifiable tryptic peptide fraction in DMD that is consistent with previous reports of increased spectrin Band 2 [32P]-phosphorylation in DMD

  8. Vicilin-like peptides from Capsicum baccatum L. seeds are α-amylase inhibitors and exhibit antifungal activity against important yeasts in medical mycology.

    Science.gov (United States)

    Vieira Bard, Gabriela C; Nascimento, Viviane V; Oliveira, Antônia Elenir A; Rodrigues, Rosana; Da Cunha, Maura; Dias, Germana B; Vasconcelos, Ilka M; Carvalho, Andre O; Gomes, Valdirene M

    2014-07-01

    The objective of this study was to isolate antimicrobial peptides from Capsicum baccatum seeds and evaluate their antimicrobial activity and inhibitory effects against α-amylase. Initially, proteins from the flour of C. baccatum seeds were extracted in sodium phosphate buffer, pH 5.4, and precipitated with ammonium sulfate at 90% saturation. The D1 and D2 fractions were subjected to antifungal tests against the yeasts Saccharomyces cerevisiae, Candida albicans, Candida tropicalis, and Kluyveromyces marxiannus, and tested against α-amylases from Callosobruchus maculates and human saliva. The D2 fraction presented higher antimicrobial activity and was subjected to further purification and seven new different fractions (H1-H7) were obtained. Peptides in the H4 fraction were sequenced and the N-terminal sequences revealed homology with previously reported storage vicilins from seeds. The H4 fraction exhibited strong antifungal activity and also promoted morphological changes in yeast, including pseudohyphae formation. All fractions, including H4, inhibited mammalian α-amylase activity but only the H4 fraction was able to inhibit C. maculatus α-amylase activity. These results suggest that the fractions isolated from the seeds of C. baccatum can act directly in plant defenses against pathogens and insects. © 2014 Wiley Periodicals, Inc.

  9. Diurnal Variations in Serum Glucose, Insulin and C-Peptide of Normal Korean Adults

    International Nuclear Information System (INIS)

    Choi, Du Hyok; Chung, June Key; Lee, Hong Kyu; Koh, Chang Soon; Hong, Kee Suk

    1983-01-01

    , Group I and Group II showed 3.50±1.85 and 1.66±0.53 ng/ml of mean±S.D., respectively. Group II showed peaks parallel to those for insulin level. None out of seven in Group I showed expected increase in C-peptide increased in 5 subjects out of seven in Group I at 11 : 00 p.m. when insulin did not increase. 4. According to the integrated concentration method for a measurement of 24 hour total insulin secretion rate, the mean±S.D. of Group I was 70.4± 15.2 U and that of Group II was 58.6±21.1 U. The above results confirm that Koreans, when given ordinary diet of 2,100 kcal and 69% sugar, show insulin secretion pattern essentially similar to that of Westerners. On the contrary, when they are put on high-calorie diet of 3, 100 kcal a day, 75% of which is sugar, insulin secretion can be increased before lunch without increase in blood glucose. These results implies that insulin secretion can be affected by some other factors. The observation that an increase in C-peptide after 11 : 00 p.m. independent of insulin level supports an assertion that insulin secretion and C-peptide secretion can be thought as being physiologically dissociable, and these changes of diurnal patterns in the levels of serum insulin and C-peptide are thought to be resulted form the large meal and high-carbohydrate diet.

  10. Diurnal Variations in Serum Glucose, Insulin and C-Peptide of Normal Korean Adults

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Du Hyok; Chung, June Key; Lee, Hong Kyu; Koh, Chang Soon [Seoul National University College of Medicine, Seoul (Korea, Republic of); Hong, Kee Suk [Korea Cancer Center Hospital, Seoul (Korea, Republic of)

    1983-03-15

    , Group I and Group II showed 3.50+-1.85 and 1.66+-0.53 ng/ml of mean+-S.D., respectively. Group II showed peaks parallel to those for insulin level. None out of seven in Group I showed expected increase in C-peptide increased in 5 subjects out of seven in Group I at 11 : 00 p.m. when insulin did not increase. 4. According to the integrated concentration method for a measurement of 24 hour total insulin secretion rate, the mean+-S.D. of Group I was 70.4+- 15.2 U and that of Group II was 58.6+-21.1 U. The above results confirm that Koreans, when given ordinary diet of 2,100 kcal and 69% sugar, show insulin secretion pattern essentially similar to that of Westerners. On the contrary, when they are put on high-calorie diet of 3, 100 kcal a day, 75% of which is sugar, insulin secretion can be increased before lunch without increase in blood glucose. These results implies that insulin secretion can be affected by some other factors. The observation that an increase in C-peptide after 11 : 00 p.m. independent of insulin level supports an assertion that insulin secretion and C-peptide secretion can be thought as being physiologically dissociable, and these changes of diurnal patterns in the levels of serum insulin and C-peptide are thought to be resulted form the large meal and high-carbohydrate diet.

  11. Levels of saliva cotinine in electronic cigarette users.

    Science.gov (United States)

    Etter, Jean-François

    2014-05-01

    To assess saliva cotinine levels in experienced users of e-cigarettes ('vapers'). An internet survey in 2011 and 2012, with collection of saliva vials by mail. Participants were 71 users of e-cigarettes enrolled mainly on websites and online forums dedicated to e-cigarettes. Use of e-cigarettes, tobacco and nicotine medications. Collection of saliva by mail and analysis of cotinine by liquid chromatography-mass spectrometry. Most participants (89%) were former smokers, most (92%) were using e-cigarettes daily, had been using e-cigarettes for 12 months on average and puffed a median of 150 times per day on their e-cigarettes [mean = 220 puffs/day, 95% confidence interval (CI) = 169-271]. The median concentration of nicotine in refill liquids was 16 mg/ml (mean = 16.4, 95% CI = 14.5-18.3). In the 62 e-cigarette users who, in the past 5 days, had not used any tobacco or nicotine medications, the median cotinine level was 353 ng/ml (mean = 374, 95% CI = 318-429), the correlation between cotinine and nicotine concentration in e-liquids was r = 0.33 (P = 0.013), and the correlation between cotinine and the number of cigarettes smoked per day before quitting smoking was r = 0.48 (P Addiction.

  12. Investigation of mixed saliva by optoelectronic methods

    Science.gov (United States)

    Savchenko, Ekaterina; Nepomnyashchaya, Elina; Baranov, Maksim; Velichko, Elena; Aksenov, Evgenii; Bogomaz, Tatyana

    2018-04-01

    At present, saliva and its properties are being actively studied. Human saliva is a unique biological material that has potential in clinical practice. A detailed analysis of the characteristics and properties of saliva is relevant for diagnostic purposes. In this paper, the properties and characteristics of saliva are studied using optoelectronic methods: dynamic light scattering, electrophoretic light scattering and optical microscopy. Mixed saliva from a healthy patient and patient with diabetes mellitus type 2 was used as an object of the study. The dynamics of the behavior of a healthy and patient with diabetes mellitus type 2 is visible according to the results obtained. All three methods confirm hypothesis of structural changes in mixed saliva in the disease of diabetes mellitus type 2.

  13. Measurement of plasma canine C peptide by radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Besch, W; Woltanski, K P; Fischer, U; Kohnert, K D; Ziegler, M

    1985-12-01

    A sensitive radioimmunoassay for canine C peptide (CCP) was established using synthetic CCP, a specific antiserum, and rabbit anti-guinea pig serum. Radioiodination was performed according to a modified chloramine T method. Tracer preparations have been used for 6 weeks after iodination. The standard curve ranges from 0.028 to 3.0 nmol/l. The intra-assay coefficient of variation (CV) was 3-5% and the inter-assay CV was 6-9% in the optimal range between 0.3 and 0.8 nmol/l. The average recovery of CCP added to plasma samples was 100.6% (n = 9). Canine insulin, porcine proinsulin, bovine proinsulin, and human C peptide exhibited no cross-reactivity. The mean fasting plasma CCP concentration was 0.089 +- 0.021 nmol/l in normal dogs and -0.005 +- 0.007 nmol/l (mean +- SEM) in diabetic dogs, respectively.

  14. Enhancement of Cellulose Degradation by Cattle Saliva

    Science.gov (United States)

    Seki, Yasutaka; Kikuchi, Yukiko; Kimura, Yoshihiro; Yoshimoto, Ryo; Takahashi, Masatoshi; Aburai, Kenichi; Kanai, Yoshihiro; Ruike, Tatsushi; Iwabata, Kazuki; Sugawara, Fumio; Sakai, Hideki; Abe, Masahiko; Sakaguchi, Kengo

    2015-01-01

    Saccharification of cellulose is a promising technique for producing alternative source of energy. However, the efficiency of conversion of cellulose into soluble sugar using any currently available methodology is too low for industrial application. Many additives, such as surfactants, have been shown to enhance the efficiency of cellulose-to-sugar conversion. In this study, we have examined first whether cattle saliva, as an additive, would enhance the cellulase-catalyzed hydrolysis of cellulose, and subsequently elucidated the mechanism by which cattle saliva enhanced this conversion. Although cattle saliva, by itself, did not degrade cellulose, it enhanced the cellulase-catalyzed degradation of cellulose. Thus, the amount of reducing sugar produced increased approximately 2.9-fold by the addition of cattle saliva. We also found that non-enzymatic proteins, which were present in cattle saliva, were responsible for causing the enhancement effect. Third, the mechanism of cattle saliva mediated enhancement of cellulase activity was probably similar to that of the canonical surfactants. Cattle saliva is available in large amounts easily and cheaply, and it can be used without further purification. Thus, cattle saliva could be a promising additive for efficient saccharification of cellulose on an industrial scale. PMID:26402242

  15. MDR1 P-glycoprotein transports endogenous opioid peptides

    NARCIS (Netherlands)

    Oude Elferink, R. P.; Zadina, J.

    2001-01-01

    MDR1 P-glycoprotein is generally regarded as an efflux pump for amphipathic toxic compounds. The question remains, however, whether certain endogenous compounds are also substrates for this transporter. Certain peptides have been shown to interact with MDR1 Pgp as well and we have therefore

  16. pH-dependence of the specific binding of Cu(II) and Zn(II) ions to the amyloid-β peptide

    International Nuclear Information System (INIS)

    Ghalebani, Leila; Wahlström, Anna; Danielsson, Jens; Wärmländer, Sebastian K.T.S.; Gräslund, Astrid

    2012-01-01

    Highlights: ► Cu(II) and Zn(II) display pH-dependent binding to the Aβ(1–40) peptide. ► At pH 7.4 both metal ions display residue-specific binding to the Aβ peptide. ► At pH 5.5 the binding specificity is lost for Zn(II). ► Differential Cu(II) and Zn(II) binding may help explain metal-induced AD toxicity. -- Abstract: Metal ions like Cu(II) and Zn(II) are accumulated in Alzheimer’s disease amyloid plaques. The amyloid-β (Aβ) peptide involved in the disease interacts with these metal ions at neutral pH via ligands provided by the N-terminal histidines and the N-terminus. The present study uses high-resolution NMR spectroscopy to monitor the residue-specific interactions of Cu(II) and Zn(II) with 15 N- and 13 C, 15 N-labeled Aβ(1–40) peptides at varying pH levels. At pH 7.4 both ions bind to the specific ligands, competing with one another. At pH 5.5 Cu(II) retains its specific histidine ligands, while Zn(II) seems to lack residue-specific interactions. The low pH mimics acidosis which is linked to inflammatory processes in vivo. The results suggest that the cell toxic effects of redox active Cu(II) binding to Aβ may be reversed by the protective activity of non-redox active Zn(II) binding to the same major binding site under non-acidic conditions. Under acidic conditions, the protective effect of Zn(II) may be decreased or changed, since Zn(II) is less able to compete with Cu(II) for the specific binding site on the Aβ peptide under these conditions.

  17. Effect of gestational age and blood glucose on C-peptide excretion rate and clearance in neonates.

    Science.gov (United States)

    Salis, Emma R; Soelbeck, Mikkel K; Reith, David M; Wheeler, Benjamin J; Broadbent, Roland S; Medlicott, Natalie J

    2016-01-01

    The aim of this study was to measure urinary C-peptide concentrations, and then calculate C-peptide clearance (Cl), and excretion rate (UER) in neonates. In addition, the effect of gestational age (GA) and blood glucose levels (BGL) on C-peptide UER were investigated. Insulin concentrations in plasma and C-peptide concentrations were measured in plasma and urine, in 20 neonates. Chemiluminescent immunoassays were used for insulin and C-peptide measurements, with urine diluted to 40% with bovine serum albumin 1% in phosphate buffered saline. Urine volume and time of collection were recorded and used to calculate UER and Cl. The mean Cl of C-peptide was 0.309 ± 0.329 mL/min/kg, and UER was 0.0329 ± 0.0342 pmol/min/kg. Correlations between Cl or UER and GA were not significant (P > 0.05). No significant correlation was shown between Cl or UER and BGL (P > 0.05). Both Cl and UER were highly variable in neonates, but were not correlated with GA. Additionally, BGL did not appear to affect C-peptide UER and Cl. As GA and BGL did not appear to affect Cl and UER, urinary C-peptide may provide a non-invasive method of measuring insulin production in neonates. © 2015 The Authors. Journal of Paediatrics and Child Health © 2015 Paediatrics and Child Health Division (Royal Australasian College of Physicians).

  18. Nature of the Charged-Group Effect on the Stability of the C-Peptide Helix

    Science.gov (United States)

    Shoemaker, Kevin R.; Kim, Peter S.; Brems, David N.; Marqusee, Susan; York, Eunice J.; Chaiken, Irwin M.; Stewart, John M.; Baldwin, Robert L.

    1985-04-01

    The residues responsible for the pH-dependent stability of the helix formed by the isolated C-peptide (residues 1-13 of ribonuclease A) have been identified by chemical synthesis of analogues and measurement of their helix-forming properties. Each of the residues ionizing between pH 2 and pH 8 has been replaced separately by an uncharged residue. Protonation of Glu-2- is responsible for the sharp decrease in helix stability between pH 5 and pH 2, and deprotonation of His-12+ causes a similar decrease between pH 5 and pH 8. Glu-9- is not needed for helix stability. The results cannot be explained by the Zimm-Bragg model and host-guest data for α -helix formation, which predict that the stability of the C-peptide helix should increase when Glu-2- is protonated or when His-12+ is deprotonated. Moreover, histidine+ is a strong helix-breaker in host-guest studies. In proteins, acidic and basic residues tend to occur at opposite ends of α -helices: acidic residues occur preferentially near the NH2-terminal end and basic residues near the COOH-terminal end. A possible explanation, based on a helix dipole model, has been given [Blagdon, D. E. & Goodman, M. (1975) Biopolymers 14, 241-245]. Our results are consistent with the helix dipole model and they support the suggestion that the distribution of charged residues in protein helices reflects the helix-stabilizing propensity of those residues. Because Glu-9 is not needed for helix stability, a possible Glu-9-\\cdots His-12+ salt bridge does not contribute significantly to helix stability. The role of a possible Glu-2-\\cdots Arg-10+ salt bridge has not yet been evaluated. A charged-group effect on α -helix stability in water has also been observed in a different peptide system [Ihara, S., Ooi, T. & Takahashi, S. (1982) Biopolymers 21, 131-145]: block copolymers containing (Ala)20 and (Glu)20 show partial helix formation at low temperatures, pH 7.5, where the glutamic acid residues are ionized. (Glu)20(Ala)20Phe forms a

  19. Oral health status, salivary pH status, and Streptococcus mutans counts in dental plaques and saliva of children with acute lymphoblastic leukemia.

    Science.gov (United States)

    Mazaheri, Romina; Jabbarifar, Ebrahim; Ghasemi, Elnaz; Akkafzadeh, Elahe; Poursaeid, Elmira

    2017-01-01

    Acute lymphoblastic leukemia (ALL), accounting for 23% of all malignancies in children, is the most prevalent pediatric malignancy. This study compared dental caries, oral hygiene status, salivary pH, and Streptococcus mutans counts in dental plaques and saliva of children with leukemia with those of healthy controls. This case-control cross-sectional study assessed 32 children with ALL and 32 healthy children (4-9-year-old) for gingival bleeding index (GBI), decayed, missing, and filled/decayed, missing, and filled surfaces (DMF/dmfs), and plaque index (PI). Sampling was performed to determine salivary pH and S. mutans counts of the participants. The two groups matched in terms of age, gender, and socioeconomic status. The groups were compared using independent t-test, Mann-Whitney test, Chi-square test, and Spearman's and Pearson's correlation analyses. The mean DMF/dmfs and GBI were significantly higher in the ALL group (P DMF / dmfs = 0.03; P GBI = 0.04). However, the two groups were not significantly different in the mean PI values ( P = 0.47). The mean S. mutans counts in dental plaques and saliva of the children with leukemia were significantly lower than the healthy controls ( P salivary pH was significantly lower in the ALL group compared to the control group ( P salivary pH, and cumulative effects of other risk factors highlight the significance of oral hygiene training programs (for the parents of these children) and regular dental examinations for these children.

  20. Kinetics of antibody response in BALB/c and C57BL/6 mice bitten by Phlebotomus papatasi.

    Directory of Open Access Journals (Sweden)

    Michaela Vlkova

    Full Text Available BACKGROUND: Phlebotomine sand flies are blood-sucking insects transmitting Leishmania parasites. In bitten hosts, sand fly saliva elicits specific immune response and the humoral immunity was shown to reflect the intensity of sand fly exposure. Thus, anti-saliva antibodies were suggested as the potential risk marker of Leishmania transmission. In this study, we examined the long-term kinetics and persistence of anti-Phlebotomus papatasi saliva antibody response in BALB/c and C57BL/6 mice. We also tested the reactivity of mice sera with P. papatasi salivary antigens and with the recombinant proteins. METHODOLOGY/PRINCIPAL FINDINGS: Sera of BALB/c and C57BL/6 mice experimentally bitten by Phlebotomus papatasi were tested by ELISA for the presence of anti-saliva IgE, IgG and its subclasses. We detected a significant increase of specific IgG and IgG1 in both mice strains and IgG2b in BALB/c mice that positively correlated with the number of blood-fed P. papatasi females. Using western blot and mass spectrometry we identified the major P. papatasi antigens as Yellow-related proteins, D7-related proteins, antigen 5-related proteins and SP-15-like proteins. We therefore tested the reactivity of mice sera with four P. papatasi recombinant proteins coding for most of these potential antigens (PpSP44, PpSP42, PpSP30, and PpSP28. Each mouse serum reacted with at least one of the recombinant protein tested, although none of the recombinant proteins were recognized by all sera. CONCLUSIONS: Our data confirmed the concept of using anti-sand fly saliva antibodies as a marker of sand fly exposure in Phlebotomus papatasi-mice model. As screening of specific antibodies is limited by the availability of salivary gland homogenate, utilization of recombinant proteins in such studies would be beneficial. Our present work demonstrates the feasibility of this implementation. A combination of recombinant salivary proteins is recommended for evaluation of intensity of

  1. Upregulation of α7 Nicotinic Receptors by Acetylcholinesterase C-Terminal Peptides

    Science.gov (United States)

    Bond, Cherie E.; Zimmermann, Martina; Greenfield, Susan A.

    2009-01-01

    Background The alpha-7 nicotinic acetylcholine receptor (α7-nAChR) is well known as a potent calcium ionophore that, in the brain, has been implicated in excitotoxicity and hence in the underlying mechanisms of neurodegenerative disorders such as Alzheimer's disease. Previous research implied that the activity of this receptor may be modified by exposure to a peptide fragment derived from the C-terminal region of the enzyme acetylcholinesterase. This investigation was undertaken to determine if the functional changes observed could be attributed to peptide binding interaction with the α7-nAChR, or peptide modulation of receptor expression. Methodology/Principal Findings This study provides evidence that two peptides derived from the C-terminus of acetylcholinesterase, not only selectively displace specific bungarotoxin binding at the α7-nAChR, but also alter receptor binding properties for its familiar ligands, including the alternative endogenous agonist choline. Of more long-term significance, these peptides also induce upregulation of α7-nAChR mRNA and protein expression, as well as enhancing receptor trafficking to the plasma membrane. Conclusions/Significance The results reported here demonstrate a hitherto unknown relationship between the α7-nAChR and the non-enzymatic functions of acetylcholinesterase, mediated independently by its C-terminal domain. Such an interaction may prove valuable as a pharmacological tool, prompting new approaches for understanding, and combating, the process of neurodegeneration. PMID:19287501

  2. Dessert formulation using sucralose and dextrin affects favorably postprandial response to glucose, insulin, and C-peptide in type 2 diabetic patients.

    Science.gov (United States)

    Argyri, Konstantina; Sotiropoulos, Alexios; Psarou, Eirini; Papazafiropoulou, Athanasia; Zampelas, Antonios; Kapsokefalou, Maria

    2013-01-01

    Dessert compositions may conform to diabetic diet when it contains low sugar or artificial sweetener to replace sugar. However, it is still questionable whether glycemic control in type 2 diabetes patients is improved by the use of diet-conforming dessert compositions. To compare, in type 2 diabetes patients, the glycemic, insulin, and C-peptide responses to seven modified dessert compositions for diabetics (D-dessert) with the response to seven similar desserts of non-modified composition, used as control desserts (C-dessert). Seventy type 2 diabetes patients were allocated to seven groups of ten. On three occasions, each patient received either the meal which consisted of bread and cheese, or the meal and D-dessert, or the meal and the respective C-dessert. Differences in postprandial glucose, insulin, and C-peptide were evaluated using analysis of repeated measures at 0, 30, 60, 90, and 120 min after consumption. D-cake and D-pastry cream resulted in lower glucose levels (8.81 ± 0.32 mmol/l and 8.67 ± 0.36 mmol/l, respectively) and D-strawberry jelly in lower insulin levels (16.46 ± 2.66 μU/ml) than the respective C-desserts (9.99 ± 0.32 mmol/l for C-cake, 9.28 ± 0.36 mmol/l for C-pastry cream, and 27.42 ± 2.66 μU/ml for C-strawberry jelly) (p pastry cream increased glucose to a lesser extent than C-pastry cream (p < 0.05). Similar effects were reported for D-milk dessert, D-millefeuille, and D-chocolate on glucose, insulin, and C-peptide at specific timepoints. D-crème caramel showed no effect. Some desserts formulated with sugar substitutes and soluble fiber may have a favorable effect on postprandial levels of glucose, insulin, and C-peptide in type 2 diabetic patients.

  3. Changes in the Concentration of Ions in Saliva and Dental Plaque after Application of CPP-ACP with and without Fluoride among 6-9 Year Old Children.

    Science.gov (United States)

    Poureslami, H; Hoseinifar, Ra; Khazaeli, P; Hoseinifar, Re; Sharifi, H; Poureslami, P

    2017-03-01

    The casein phospho peptide-amorphous calcium phosphate with or without fluoride (CPP-ACPF and CPP-ACP respectively) are of considerably new materials which are highly recommended for prevention of dental caries. However, there is a shortage in literature on how they affect the ion concentration of saliva or dental plaque. The aim of this study was to evaluate the concentration of calcium, phosphate and fluoride in the plaque and saliva of children with Early Childhood Caries (ECC) after applying the CPP-ACP paste in comparison with the use of CPP-ACPF paste. One ml of un-stimulated saliva of 25 preschool children was collected and then 1 mg of the plaque sample was collected from the buccal surfaces of the two first primary molars on the upper jaw. CPP-ACP as well as CPP-ACPF pastes were applied on the tooth surfaces in two separate steps. In steps, plaque and saliva sampling was performed after 60 minutes. The amount of calcium ions was measured by Atomic Absorption Device and the amount of phosphate and fluoride ions was measured by Ion Chromatography instrument. Data were analyzed using Repeated Measurements ANOVA at a p Application of both CPP-ACPF and CPP-ACP significantly increased the concentration of calcium, phosphate, and fluoride in both saliva and dental plaque. Moreover, significantly higher salivary fluoride concentration was seen after application of CPP-ACPF compared to CPP-ACP. No other significant difference was observed between these two materials. CPP-ACPF can be more useful than CPP-ACP in protecting the primary teeth against caries process, especially when there is poor hygiene.

  4. Corrosion in artificial saliva of a Ni-Cr-based dental alloy joined by TIG welding and conventional brazing.

    Science.gov (United States)

    Matos, Irma C; Bastos, Ivan N; Diniz, Marília G; de Miranda, Mauro S

    2015-08-01

    Fixed prosthesis and partial dental prosthesis frameworks are usually made from welded Ni-Cr-based alloys. These structures can corrode in saliva and have to be investigated to establish their safety. The purpose of this study was to evaluate the corrosion behavior of joints joined by tungsten inert gas (TIG) welding and conventional brazing in specimens made of commercial Ni-Cr alloy in Fusayama artificial saliva at 37°C (pH 2.5 and 5.5). Eighteen Ni-Cr base metal specimens were cast and welded by brazing or tungsten inert gas methods. The specimens were divided into 3 groups (base metal, 2 welded specimens), and the composition and microstructure were qualitatively evaluated. The results of potential corrosion and corrosion current density were analyzed with a 1-way analysis of variance and the Tukey test for pairwise comparisons (α=.05). Base metal and tungsten inert gas welded material showed equivalent results in electrochemical corrosion tests, while the air-torched specimens exhibited low corrosion resistance. The performance was worst at pH 2.5. These results suggest that tungsten inert gas is a suitable welding process for use in dentistry, because the final microstructure does not reduce the corrosion resistance in artificial saliva at 37°C, even in a corrosion-testing medium that facilitates galvanic corrosion processes. Moreover, the corrosion current density of brazed Ni-Cr alloy joints was significantly higher (P<.001) than the base metal and tungsten inert gas welded joints. Copyright © 2015 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

  5. Effect of saliva contamination and artificial aging on different primer/cement systems bonded to zirconia.

    Science.gov (United States)

    Pitta, João; Branco, Teresa C; Portugal, Jaime

    2018-05-01

    Saliva contamination has been shown to decrease bonding to zirconia. Adopting a less contamination-sensitive cement system may be an alternative to decontamination. The purpose of this in vitro study was to assess the ability of different primer/cement systems to promote a durable bond to zirconia after saliva contamination. Zirconia blocks (Lava Plus) (N=320) were airborne-particle abraded (50 μm Al 2 O 3 ) and divided into 32 experimental groups (n=10) according to the variables in the study: saliva contamination; primer/cement system (Panavia SA [PSA]; RelyX Unicem 2 [RU2]; Bifix SE [BSE]; Panavia F2.0 [PF2]; Scotchbond Universal + RelyX Ultimate [SBU+RXU]; Futurabond M+ + Bifix QM [FBM+BQM]; All-Bond Universal + Duo-link [ABU+DL]; Z-Prime Plus + Duo-link [ZPP+DL]; and aging period (72 hours; 30 days with 10 000 thermocycles at 5°C to 55°C). After half of the blocks had been contaminated with fresh human saliva for 10 minutes, rinsed with water, and air-dried, each primer/cement was applied. Polymerized composite resin disks were then placed over the cement, and the resin cement was light-polymerized for 20 seconds each at 2 opposite margins. After the aging time, the specimens were tested in shear (1 mm/min). The failure mode was classified as adhesive, cohesive, or mixed. Statistical analysis of the shear bond strength (SBS) data was performed with ANOVA followed by Tukey honest significant difference post hoc tests. Chi-square tests were used to analyze the failure mode data (α=.05). The mean SBS ranged between 4.2 and 34.5 MPa. Shear bond strength was influenced (Pcontamination, aging time). SBU+RXU and FBM+BQM showed a higher mean SBS than those of the other experimental groups (Pcontamination (P>.05). Failure was predominantly classified as adhesive. In general, saliva contamination and aging decreased bonding efficacy. Two systems, combining an application of a universal adhesive and a resin cement (SBU+RXU and FBM+BQM) were not affected by

  6. Role of C-peptide in Altered Lipid Profile among Apparently Healthy Adults of Vijayapura City, Karnataka

    Directory of Open Access Journals (Sweden)

    Chandrahas M.Kulkarni

    2016-04-01

    Full Text Available Background: C-peptide is produced in equimolar concentration during insulin production as inactive molecule by beta islet cells of Langerhans. C-peptide is most useful biomarker of endogenous insulin production. Aim and Objectives: To predict metabolic syndrome in advance by estimation of C-peptide and lipid profile in healthy adults. Material and Methods: Serum C-peptide, fasting blood glucose and lipid profile of 128 healthy individuals were estimated. Adults in the age group of 18 to 60 years of both sexes were included in study. Results: C-peptide levels were increased in 27%, Serum cholesterol in 30%, LDL Cholesterol in 55% and triglyceride levels in 21% of healthy individuals. Significant correlation was observed between C peptide, age, serum cholesterol, LDL and cholesterol LDL ratio in male subjects only. In our study group most of the subjects (both males and females fell in overweight group. Conclusion: Cpeptide level and lipid profile may be considered as useful biomarkers to predict type 2 diabetes mellitus in advance, possibly due to insulin resistance.

  7. Insulin and C-peptide secretion in non-obese patients with polycystic ovarian disease.

    Science.gov (United States)

    Mahabeer, S; Jialal, I; Norman, R J; Naidoo, C; Reddi, K; Joubert, S M

    1989-09-01

    Plasma glucose, immunoreactive insulin (IRI) and C-peptide responses during an oral glucose tolerance test (oGTT) were assessed in 11 non-obese patients with polycystic ovarian disease (PCOD) and 11 reference subjects matched for age, height and weight. Also, 6 patients with PCOD and 6 normal women were subjected to intravenous glucose tolerance testing (ivGTT) On oGTT, all subjects exhibited normal glucose tolerance; however, PCOD patients had significantly higher mean plasma glucose levels at 30, 60, 90 and 120 min and higher mean incremental glucose areas. In addition the patients with polycystic ovaries showed higher mean basal IRI and C-peptide levels, higher mean glucose stimulated IRI and C-peptide levels and higher mean incremental IRI and C-peptide values. The molar ratios of C-peptide/IRI were significantly lower in the PCOD group at all time intervals after glucose stimulation when compared to the normal women. During ivGTT, there were significantly higher mean glucose levels at 5, 40, 50 and 60 min in the PCOD group when compared to the reference group. The IRI response to intravenous glucose in the PCOD women was similar to the reference group. The findings on oGTT suggest that non-obese patients with PCOD have increased pancreatic IRI secretion as well as impaired hepatic extraction of the hormone.

  8. Peptide-targeted delivery of a pH sensor for quantitative measurements of intraglycosomal pH in live Trypanosoma brucei.

    Science.gov (United States)

    Lin, Sheng; Morris, Meredith T; Ackroyd, P Christine; Morris, James C; Christensen, Kenneth A

    2013-05-28

    Studies of dynamic changes in organelles of protozoan parasite Trypanosoma brucei have been limited, in part because of the difficulty of targeting analytical probes to specific subcellular compartments. Here we demonstrate application of a ratiometric probe for pH quantification in T. brucei glycosomes. The probe consists of a peptide encoding the peroxisomal targeting sequence (F-PTS1, acetyl-CKGGAKL) coupled to fluorescein, which responds to pH. When incubated with living parasites, the probe is internalized within vesicular structures that colocalize with a glycosomal marker. Inhibition of uptake of F-PTS1 at 4 °C and pulse-chase colocalization with fluorescent dextran suggested that the probe is initially taken up by non-receptor-mediated endocytosis but is subsequently transported separately from dextran and localized within glycosomes, prior to the final fusion of labeled glycosomes and lysosomes as part of glycosomal turnover. Intraorganellar measurements and pH calibration with F-PTS1 in T. brucei glycosomes indicate that the resting glycosomal pH under physiological conditions is 7.4 ± 0.2. However, incubation in glucose-depleted buffer triggered mild acidification of the glycosome over a period of 20 min, with a final observed pH of 6.8 ± 0.3. This glycosomal acidification was reversed by reintroduction of glucose. Coupling of ratiometric fluorescent sensors and reporters to PTS peptides offers an invaluable tool for monitoring in situ glycosomal response(s) to changing environmental conditions and could be applied to additional kinetoplastid parasites.

  9. Detection of phencyclidine usage by radioimmunoassay of saliva

    International Nuclear Information System (INIS)

    McCarron, M.M.; Walberg, C.B.; Soares, J.R.; Gross, S.J.; Baselt, R.C.

    1984-01-01

    Paired serum and saliva samples, obtained from 100 emergency department patients suspected of phencyclidine (PCP) intoxication, were analyzed using a specific PCP radioimmunoassay (RIA). Seventy-four of the 100 saliva samples and 75 of the paired serum samples were positive for PCP. The final clinical diagnosis was PCP intoxication in 79 cases. Of these, both serum and saliva tests were positive in 70 cases, only serum was positive in two cases, and both serum and saliva samples were negative in seven cases. The concentration of PCP in the samples did not correlate with the severity of PCP intoxication. In the remaining 21 cases, with no clinical evidence of PCP intoxication, PCP assays were negative in both serum and saliva in 17 cases, three patients had positive saliva and serum tests, and one other patient had a positive PCP saliva assay. Thus, saliva would appear to be as reliable as serum as a specimen for PCP analysis

  10. Single administration of p2TA (AB103, a CD28 antagonist peptide, prevents inflammatory and thrombotic reactions and protects against gastrointestinal injury in total-body irradiated mice.

    Directory of Open Access Journals (Sweden)

    Salida Mirzoeva

    Full Text Available The goal of this study was to elucidate the action of the CD28 mimetic peptide p2TA (AB103 that attenuates an excessive inflammatory response in mitigating radiation-induced inflammatory injuries. BALB/c and A/J mice were divided into four groups: Control (C, Peptide (P; 5 mg/kg of p2TA peptide, Radiation (R; total body irradiation with 8 Gy γ-rays, and Radiation + Peptide (RP; irradiation followed by p2TA peptide 24 h later. Gastrointestinal tissue damage was evaluated by analysis of jejunum histopathology and immunohistochemistry for cell proliferation (Cyclin D1 and inflammation (COX-2 markers, as well as the presence of macrophages (F4/80. Pro-inflammatory cytokines IL-6 and KC as well as fibrinogen were quantified in plasma samples obtained from the same mice. Our results demonstrated that administration of p2TA peptide significantly reduced the irradiation-induced increase of IL-6 and fibrinogen in plasma 7 days after exposure. Seven days after total body irradiation with 8 Gy of gamma rays numbers of intestinal crypt cells were reduced and villi were shorter in irradiated animals compared to the controls. The p2TA peptide delivery 24 h after irradiation led to improved morphology of villi and crypts, increased Cyclin D1 expression, decreased COX-2 staining and decreased numbers of macrophages in small intestine of irradiated mice. Our study suggests that attenuation of CD28 signaling is a promising therapeutic approach for mitigation of radiation-induced tissue injury.

  11. Variables associated with persistence of C-Peptide secretion among patients with Type 1 diabetes mellitus

    Directory of Open Access Journals (Sweden)

    Ibrahim Abbood Zaboon

    2017-01-01

    Full Text Available Background: C-peptide is a reliable method for estimating the beta-cell residual function. The objective of this study to assess the variables associated with persistence of C-peptide secretion among patients with Type 1 diabetes mellitus (T1DM. Patients and Methods: This was a cross-sectional study conducted from October 2015 to September 2016. This study enrolled patients with T1DM with at least 1 year or more duration. Random C-peptide with concomitant plasma glucose at least 144 mg/dl (8 mmol/l was measured and at this cutoff considered as a stimulated value. Variables that were assessed were age at the time of enrollment, age at the diagnosis of diabetes, gender, family history of diabetes, duration of diabetes, frequency of insulin per day, insulin dose (units/kg/day, type of insulin, devices delivery, body mass index (BMI at enrollment, blood pressure, glucose (plasma, lipid profile, glycated hemoglobin (HbA1c, thyrotropin (TSH, and antibodies to glutamic acid decarboxylase (GAD65, thyroid peroxidase antibodies (anti-TPO, and tissue transglutaminase antibodies-IgA (anti-TTG-IgA. Results: A total 324 patients were included in the study. A higher level of C-peptide has been seen if the disease acquired at the age of 18 years and older with detectable C-peptide observed among 17.7% of those diagnosed at age <18 years versus 31.7% for those aged 18 years or above. The more the duration of diabetes, the more is the loss of C-peptide. On logistic regression analysis, only duration of diabetes <6 years, and insulin dose <1 U/kg/day were statistically significantly associated with the detectable level of C-peptide in this cohort of T1DM. Conclusion: Diagnosis of TIDM at a late age, positive family history of diabetes, those requiring <1 U of insulin per kg per day, and higher fasting glucose was associated with higher and more detectable C-peptide. On multivariable analysis, the only duration of diabetes <6 years and insulin dose <1 U of insulin

  12. C-Peptide Is a Sensitive Indicator for the Diagnosis of Metabolic Syndrome in Subjects from Central Mexico.

    Science.gov (United States)

    Gonzalez-Mejia, M Elba; Porchia, Leonardo M; Torres-Rasgado, Enrique; Ruiz-Vivanco, Guadalupe; Pulido-Pérez, Patricia; Báez-Duarte, Blanca G; Pérez-Fuentes, Ricardo

    2016-05-01

    Metabolic Syndrome (MetS) is associated with elevated risk for developing diabetes and cardiovascular disease. A key component of MetS is the development of insulin resistance (IR). The homeostatic model assessment (HOMA) model can determine IR by using insulin or C-peptide concentrations; however, the efficiency of insulin and C-peptide to determine MetS has not been compared. The aim of the study was to compare the efficiency of C-peptide and insulin to determine MetS in Mexicans. Anthropometrics, glucose, insulin, C-peptide, triglycerides, and high-density lipoproteins were determined in 156 nonpregnant females and 114 males. Subjects were separated into normal or positive for MetS. IR was determined by the HOMA2 calculator using insulin or C-peptide. Correlations were calculated using the Spearman correlation coefficient (ρ). Differences between correlations were determined by calculating Steiger's Z. The sensitivity was determined by the area under receiver operating characteristics curve (AUC) analysis. Independent of the MetS definition [Adult Treatment Panel III (ATP III), International Diabetes Federation (IDF), or World Health Organization (WHO)], C-peptide and insulin were significantly higher in MetS subjects (P indicator of MetS. Since C-peptide has recently emerged as a biomolecule with significant importance for inflammatory diseases, monitoring C-peptide levels will aid clinicians in preventing MetS.

  13. Proteomic Analysis of Saliva Identifies Potential Biomarkers for Orthodontic Tooth Movement

    Science.gov (United States)

    Ellias, Mohd Faiz; Zainal Ariffin, Shahrul Hisham; Karsani, Saiful Anuar; Abdul Rahman, Mariati; Senafi, Shahidan; Megat Abdul Wahab, Rohaya

    2012-01-01

    Orthodontic treatment has been shown to induce inflammation, followed by bone remodelling in the periodontium. These processes trigger the secretion of various proteins and enzymes into the saliva. This study aims to identify salivary proteins that change in expression during orthodontic tooth movement. These differentially expressed proteins can potentially serve as protein biomarkers for the monitoring of orthodontic treatment and tooth movement. Whole saliva from three healthy female subjects were collected before force application using fixed appliance and at 14 days after 0.014′′ Niti wire was applied. Salivary proteins were resolved using two-dimensional gel electrophoresis (2DE) over a pH range of 3–10, and the resulting proteome profiles were compared. Differentially expressed protein spots were then identified by MALDI-TOF/TOF tandem mass spectrometry. Nine proteins were found to be differentially expressed; however, only eight were identified by MALDI-TOF/TOF. Four of these proteins—Protein S100-A9, immunoglobulin J chain, Ig alpha-1 chain C region, and CRISP-3—have known roles in inflammation and bone resorption. PMID:22919344

  14. cDNA cloning and immunological characterization of the rye grass allergen Lol p I.

    Science.gov (United States)

    Perez, M; Ishioka, G Y; Walker, L E; Chesnut, R W

    1990-09-25

    The complete amino acid sequence of two "isoallergenic" forms of Lol p I, the major rye grass (Lolium perenne) pollen allergen, was deduced from cDNA sequence analysis. cDNA clones isolated from a Lolium perenne pollen library contained an open reading frame coding for a 240-amino acid protein. Comparison of the nucleotide and deduced amino acid sequence of two of these clones revealed four changes at the amino acid level and numerous nucleotide differences. Both clones contained one possible asparagine-linked glycosylation site. Northern blot analysis shows one RNA species of 1.2 kilobases. Based on the complete amino acid sequence of Lol p I, overlapping peptides covering the entire molecule were synthesized. Utilizing these peptides we have identified a determinant within the Lol p I molecule that is recognized by human leukocyte antigen class II-restricted T cells obtained from persons allergic to rye grass pollen.

  15. Quantitative analysis of differentially expressed saliva proteins in human immunodeficiency virus type 1 (HIV-1) infected individuals

    International Nuclear Information System (INIS)

    Zhang, Nawei; Zhang, Zhenyu; Feng, Shan; Wang, Qingtao; Malamud, Daniel; Deng, Haiteng

    2013-01-01

    Highlights: ► A high-throughput method for profiling and quantification of the differentially expressed proteins in saliva samples was developed. ► Identified that DMBT1, S100A7, S100A8, S100A9 and alpha defensin were up-regulated in saliva from HIV-1 seropositive patients. ► Established analytical strategies are translatable to the clinical setting. -- Abstract: In the present study, we have established a new methodology to analyze saliva proteins from HIV-1-seropositive patients before highly active antiretroviral therapy (HAART) and seronegative controls. A total of 593 and 601 proteins were identified in the pooled saliva samples from 5 HIV-1 subjects and 5 controls, respectively. Forty-one proteins were found to be differentially expressed. Bioinformatic analysis of differentially expressed salivary proteins showed an increase of antimicrobial proteins and decrease of protease inhibitors upon HIV-1 infection. To validate some of these differentially expressed proteins, a high-throughput quantitation method was established to determine concentrations of 10 salivary proteins in 40 individual saliva samples from 20 seropositive patients before HAART and 20 seronegative subjects. This method was based on limited protein separation within the zone of the stacking gel of the 1D SDS PAGE and using isotope-coded synthetic peptides as internal standards. The results demonstrated that a combination of protein profiling and targeted quantitation is an efficient method to identify and validate differentially expressed salivary proteins. Expression levels of members of the calcium-binding S100 protein family and deleted in malignant brain tumors 1 protein (DMBT1) were up-regulated while that of Mucin 5B was down-regulated in HIV-1 seropositive saliva samples, which may provide new perspectives for monitoring HIV-infection and understanding the mechanism of HIV-1 infectivity

  16. Quantitative analysis of differentially expressed saliva proteins in human immunodeficiency virus type 1 (HIV-1) infected individuals

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Nawei; Zhang, Zhenyu [Beijing Chaoyang Hospital Affiliated Capital Medical University, Beijing (China); Feng, Shan [MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing (China); Wang, Qingtao [Beijing Chaoyang Hospital Affiliated Capital Medical University, Beijing (China); Malamud, Daniel [NYU College of Dentistry, 345 East 24th Street, New York, NY 10010 (United States); Deng, Haiteng, E-mail: dht@mail.tsinghua.edu.cn [MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing (China)

    2013-04-24

    Highlights: ► A high-throughput method for profiling and quantification of the differentially expressed proteins in saliva samples was developed. ► Identified that DMBT1, S100A7, S100A8, S100A9 and alpha defensin were up-regulated in saliva from HIV-1 seropositive patients. ► Established analytical strategies are translatable to the clinical setting. -- Abstract: In the present study, we have established a new methodology to analyze saliva proteins from HIV-1-seropositive patients before highly active antiretroviral therapy (HAART) and seronegative controls. A total of 593 and 601 proteins were identified in the pooled saliva samples from 5 HIV-1 subjects and 5 controls, respectively. Forty-one proteins were found to be differentially expressed. Bioinformatic analysis of differentially expressed salivary proteins showed an increase of antimicrobial proteins and decrease of protease inhibitors upon HIV-1 infection. To validate some of these differentially expressed proteins, a high-throughput quantitation method was established to determine concentrations of 10 salivary proteins in 40 individual saliva samples from 20 seropositive patients before HAART and 20 seronegative subjects. This method was based on limited protein separation within the zone of the stacking gel of the 1D SDS PAGE and using isotope-coded synthetic peptides as internal standards. The results demonstrated that a combination of protein profiling and targeted quantitation is an efficient method to identify and validate differentially expressed salivary proteins. Expression levels of members of the calcium-binding S100 protein family and deleted in malignant brain tumors 1 protein (DMBT1) were up-regulated while that of Mucin 5B was down-regulated in HIV-1 seropositive saliva samples, which may provide new perspectives for monitoring HIV-infection and understanding the mechanism of HIV-1 infectivity.

  17. Low-molecular-weight color pI markers to monitor on-line the peptide focusing process in OFFGEL fractionation.

    Science.gov (United States)

    Michelland, Sylvie; Bourgoin-Voillard, Sandrine; Cunin, Valérie; Tollance, Axel; Bertolino, Pascal; Slais, Karel; Seve, Michel

    2017-08-01

    High-throughput mass spectrometry-based proteomic analysis requires peptide fractionation to simplify complex biological samples and increase proteome coverage. OFFGEL fractionation technology became a common method to separate peptides or proteins using isoelectric focusing in an immobilized pH gradient. However, the OFFGEL focusing process may be further optimized and controlled in terms of separation time and pI resolution. Here we evaluated OFFGEL technology to separate peptides from different samples in the presence of low-molecular-weight (LMW) color pI markers to visualize the focusing process. LMW color pI markers covering a large pH range were added to the peptide mixture before OFFGEL fractionation using a 24-wells device encompassing the pH range 3-10. We also explored the impact of LMW color pI markers on peptide fractionation labeled previously for iTRAQ. Then, fractionated peptides were separated by RP_HPLC prior to MS analysis using MALDI-TOF/TOF mass spectrometry in MS and MS/MS modes. Here we report the performance of the peptide focusing process in the presence of LMW color pI markers as on-line trackers during the OFFGEL process and the possibility to use them as pI controls for peptide focusing. This method improves the workflow for peptide fractionation in a bottom-up proteomic approach with or without iTRAQ labeling. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. C-peptide prevents SMAD3 binding to alpha promoters to inhibit collagen type IV synthesis.

    Science.gov (United States)

    Li, Yanning; Zhong, Yan; Gong, Wenjian; Gao, Xuehan; Qi, Huanli; Liu, Kun; Qi, Jinsheng

    2018-07-01

    Activation of transforming growth factor β1 (TGFB1)/SMAD3 signaling may lead to additional synthesis of collagen type IV (COL4), which is a major contributor to extracellular matrix (ECM) accumulation in diabetic nephropathy (DN). C-peptide can attenuate fibrosis to have unique beneficial effects in DN. However, whether and how C-peptide affects TGFB1/SMAD3-activated COL4 synthesis is unclear. In this study, pathological changes, expression of COL4 a1-a5 chains ( Col4a1-a5 ), COL4 distribution and protein and TGFB1 and SMAD3 protein were first assessed in a rat model of diabetes. Then, rat mesangial cells were treated with high glucose (HG) and/or C-peptide to investigate the underlying mechanism. Col4a1-a5 expression, COL4 protein and secretion, TGFB1 protein, SMAD3 nuclear translocation and binding of SMAD3 to its cognate sites in the promoters of Col4a1a2 , Col4a3a4 and Col4a5 were measured. It was found that C-peptide attenuated glomerular pathological changes and suppressed renal Col4a1 -a5 mRNA expression, COL4 protein content and TGFB1 protein content. C-peptide had a dose-dependent effect to inhibit Col4a1-a5 mRNA expression, COL4 protein content and secretion, in HG-stimulated mesangial cells. In addition, the HG-induced increase in TGFB1 protein content was significantly reduced by C-peptide. Although not apparently affecting SMAD3 nuclear translocation, C-peptide prevented SMAD3 from binding to its sites in the Col4a1a2 , Col4a3a4 and Col4a5 promoters in HG-stimulated mesangial cells. In conclusion, C-peptide could prevent SMAD3 from binding to its sites in the Col4a1a2 , Col4a3a4 and Col4a5 promoters, to inhibit COL4 generation. These results may provide a mechanism for the alleviation of fibrosis in DN by C-peptide. © 2018 Society for Endocrinology.

  19. SALIVA AS A DIAGNOSTIC FLUID

    Directory of Open Access Journals (Sweden)

    Pezelj-Ribarić Sonja

    2015-12-01

    Full Text Available Saliva is a readily available oral fluid with many functions, from digestion, maintenance of oral tissues' integrity, to caries prevention. Changes regarding its secretion may be divided into qualitative and quantitative: both of them are a consequence of certain conditions/diseases (e.g. internal factors or nutrients/drugs ingested (e.g. external factors. During the last 15 years, technological advances gave a significant momentum to utilization of saliva as a diagnostic tool. Analysis of saliva, just like the blood analysis, has two main objectives: to identify the subjects suffering from a certain disorder, and to follow the development and progress of therapy. This paper provides an overview of possibilities for the use of saliva for diagnostic purposes and gives specific examples of some clinical investigations, with the final aim to stimulate the use of this noninvasive means for the health care promotion.

  20. Preliminary findings on the correlation of saliva pH, buffering capacity, flow, Consistency and Streptococcus mutans in relation to cigarette smoking.

    Science.gov (United States)

    Voelker, Marsha A; Simmer-Beck, Melanie; Cole, Molly; Keeven, Erin; Tira, Daniel

    2013-02-01

    The purpose of this preliminary study was to examine the relationship of caries risk, salivary buffering capacity, salivary pH, salivary quality (flow, consistency) and levels of Streptococcus mutans in relation to cigarette smoking. This clinical trial consisted of 53 volunteer patients receiving care in a university based dental hygiene clinic. Participants completed a questionnaire specific to their social history in regards to tobacco use, oral health and dietary history. Measurements of unstimulated saliva were collected followed by collection of stimulated saliva samples. These samples were used to measure salivary pH, buffering capacity and Streptococcus mutans levels. The subject's smoking status was significantly associated with caries risk (p= 0.001), with 25% of the variability of caries risk attributed to smoking. The smoking status was significantly associated with buffering capacity (p=0.025), with 9% of the variability of buffering status attributed to the smoking. Associations between smoking status and salivary pH were not statistically significant. The subject's caries risk was significantly associated with buffering capacity (p= 0.001), with 25% of the variability of caries risk attributed to the buffering capacity. The subject's caries risk was significantly associated with salivary pH (p= 0.031), with 9% of the variability of caries risk attributed to the salivary pH. The Streptococcus mutans test showed no statistical significance (p>0.05) possibly due to the number and low variance in the subjects. A relationship between caries risk and smoking, buffering capacity and smoking, and stimulated salivary pH and smoking were concluded. No significance difference (p>0.05) between caries risk and salivary pH, salivary quality and smoking, S. mutans and smoking were noted from the preliminary results.

  1. Structure of a C-terminal AHNAK peptide in a 1:2:2 complex with S100A10 and an acetylated N-terminal peptide of annexin A2

    Energy Technology Data Exchange (ETDEWEB)

    Ozorowski, Gabriel [University of California, Irvine, Irvine, CA 92697-3900 (United States); University of California, Irvine, Irvine, CA 92697-3900 (United States); Milton, Saskia [University of California, Irvine, Irvine, CA 92697-3900 (United States); Luecke, Hartmut, E-mail: hudel@uci.edu [University of California, Irvine, Irvine, CA 92697-3900 (United States); University of California, Irvine, Irvine, CA 92697-3900 (United States); University of California, Irvine, Irvine, CA 92697 (United States); University of California, Irvine, Irvine, CA 92697 (United States)

    2013-01-01

    Structure of a 20-amino-acid peptide of AHNAK bound asymmetrically to the AnxA2–S100A10A heterotetramer (1:2:2 symmetry) provides insights into the atomic level interactions that govern this membrane-repair scaffolding complex. AHNAK, a large 629 kDa protein, has been implicated in membrane repair, and the annexin A2–S100A10 heterotetramer [(p11){sub 2}(AnxA2){sub 2})] has high affinity for several regions of its 1002-amino-acid C-terminal domain. (p11){sub 2}(AnxA2){sub 2} is often localized near the plasma membrane, and this C2-symmetric platform is proposed to be involved in the bridging of membrane vesicles and trafficking of proteins to the plasma membrane. All three proteins co-localize at the intracellular face of the plasma membrane in a Ca{sup 2+}-dependent manner. The binding of AHNAK to (p11){sub 2}(AnxA2){sub 2} has been studied previously, and a minimal binding motif has been mapped to a 20-amino-acid peptide corresponding to residues 5654–5673 of the AHNAK C-terminal domain. Here, the 2.5 Å resolution crystal structure of this 20-amino-acid peptide of AHNAK bound to the AnxA2–S100A10 heterotetramer (1:2:2 symmetry) is presented, which confirms the asymmetric arrangement first described by Rezvanpour and coworkers and explains why the binding motif has high affinity for (p11){sub 2}(AnxA2){sub 2}. Binding of AHNAK to the surface of (p11){sub 2}(AnxA2){sub 2} is governed by several hydrophobic interactions between side chains of AHNAK and pockets on S100A10. The pockets are large enough to accommodate a variety of hydrophobic side chains, allowing the consensus sequence to be more general. Additionally, the various hydrogen bonds formed between the AHNAK peptide and (p11){sub 2}(AnxA2){sub 2} most often involve backbone atoms of AHNAK; as a result, the side chains, particularly those that point away from S100A10/AnxA2 towards the solvent, are largely interchangeable. While the structure-based consensus sequence allows interactions with various

  2. Candida in saliva of Brazilian hemophilic patients.

    Science.gov (United States)

    Pereira, Claudio Maranhão; Pires, Fábio Ramôa; Corrêa, Maria Elvira Pizzigatti; di Hipólito Júnior, Osvaldo; Almeida, Oslei Paes de

    2004-12-01

    Hemophilia is a common hereditary hemorrhagic disorder, however little is known about the oral microflora of hemophilic patients. The aim of this study was to quantify the Candida and identify its species in non-stimulated saliva of hemophilic patients, and consider its relationship with clinical factors influencing Candida carriage. This study comprised evaluation of 86 hemophilic patients of the Hematology Center/UNICAMP and 43 healthy subjects as controls. All patients were submitted to anamnesis, intraoral examination and unstimulated saliva collection. Candida counts and species identification were performed in salivary samples. Candida was present in 64% of the hemophilic patients and in 44% of the healthy controls. C. albicans represented 65% and 68% of the isolated species, in hemophiliacs and control group respectively, and C. tropicalis was the second most common species in both groups. These results indicate that hemophilic patients carry Candida more frequently and in higher counts than healthy controls, independently of oral clinical parameter considered, as viral infections, complete dentures, transfusions of hemoderivatives, and salivary flow.

  3. Cloning of cDNAs encoding new peptides of the dermaseptin-family.

    Science.gov (United States)

    Wechselberger, C

    1998-10-14

    Dermaseptins are a group of basic (lysine-rich) peptides, 27-34 amino acids in length and involved in the defense of frog skin against microbial invasion. By using a degenerated oligonucleotide primer binding to the 5'-untranslated region of previously characterized cDNAs of these peptides, it was possible to identify new members of the dermaseptin family in the South American frogs Agalychnis annae and Pachymedusa dacnicolor. Amino acid alignment and secondary structure prediction reveals, that only five of the deduced peptides can be supposed to be also functional homologs to the known dermaseptins from Phyllomedusa bicolor and Phyllomedusa sauvagei. The remaining six peptides described in this paper have not been isolated and characterized yet.

  4. Kontrasepsi Hormonal Meningkatkan Kadar α-Amylase Saliva

    Directory of Open Access Journals (Sweden)

    Juni Handajani

    2014-06-01

    Full Text Available Salivary α-amylase atau α-amylase saliva (SAA adalah salah satu enzim dalam saliva yang berperan penting pada inisiasi digesti karbohidrat dan fungsi interaksi bakteri. Kontrasepsi hormonal sangat populer di Indonesia untuk mencegah kehamilan. Penelitian ini bertujuan untuk mengetahui kadar SAA wanita pemakai kontrasepsi pil dan suntik. Subjek penelitian sebanyak 30 perempuan usia 20-35 tahun. Prosedur penelitian telah mendapat persetujuan dari Komite Etik Fakultas Kedokteran Universitas Gadjah Mada Yogyakarta. Subjek dibagi menjadi 3 kelompok (pemakai kontrasepsi pil, suntik, dan kontrol, masing-masing 10 perempuan. Kriteria subjek antara lain subjek sehat, tidak menggunakan alat ortodontik, protesa atau mahkota, serta menggunakan kontrasepsi hormonal lebih dari 3 bulan. Sampel saliva dikumpulkan pada sore hari (16.00-18.00 WIB selama 1 menit dengan metode tanpa stimulasi. Kadar tingkat SAA diukur menggunakan ELISA kit (Salimetrics LLC dengan Optical Density (OD pada 405 nm. Data dianalisis menggunakan ANOVA (p<0,05. Hasil penelitian menunjukkan kadar SAA tertinggi pada perempuan pemakai kontrasepsi  pil dan ada perbedaan yang signifikan diantara tiga kelompok. Disimpulkan bahwa kontrasepsi hormonal meningkatkan kadar SAA. Hormonal Contraceptive Increased The Level of Salivary Α-Amylase. Salivary α-amylase (SAA is one of the most important enzymes in saliva. This enzyme was mainly involved in the initiation of the digestion of starch in the oral cavity and has significant bacterial interactive function. Hormonal contraceptives are very popular in Indonesia to avoid pregnancy. This study aimed to evaluate the level of SAA in woman who taking pill and by injection contraceptives. Thirty women were in subjects, 20-35 years old, approval ethical clearance from Ethic Committee Medical Faculty of Gadjah Mada University, Yogyakarta Indonesia. Subjects were divided into three groups (taking pill contraceptive, by injection contraceptive and

  5. Preoperative Fasting Plasma C-Peptide Levels as Predictors of Remission of Type 2 Diabetes Mellitus after Bariatric Surgery: A Systematic Review and Meta-Analysis.

    Science.gov (United States)

    Yan, Wenmao; Bai, Rixing; Yan, Ming; Song, Maomin

    2017-12-01

    The study evaluated the predictive role of preoperative fasting C-peptide, hemoglobin (Hb)A1c, fasting plasma glucose (FPG), and body mass index (BMI) levels on diabetes remission in patients with type 2 diabetes following bariatric surgery. Medline, PubMed, Central, and Google Scholar databases of up to September 7, 2016 were searched using the following terms: type 2 diabetes mellitus, gastric bypass, Roux-en-Y, anastomosis, C-peptide, weight loss, HbA/HbA1c, predictive/predictor. Meta-analysis of the pooled data indicated that fasting C-peptide was predictive of increased chance of remission of type 2 diabetes (pooled difference in means = 0.93, 95% confidence interval [CI] = 0.61 to 1.25, p fasting plasma C-peptide was associated with increased type 2 diabetes remission after bariatric surgery, whereas baseline HbA1c and FPG levels were associated with reduced chance of remission. These parameters may be used as a guideline in weighing the risks and benefits for surgical intervention in patients with type 2 diabetes.

  6. Increased (/sup 32/P)-phosphorylation of tryptic peptides of erythrocyte spectrin in Duchenne muscular dystrophy

    Energy Technology Data Exchange (ETDEWEB)

    Mabry, M.E.; Roses, A.D.

    Increased (32P)-incorporation in tryptic peptides of the erythrocyte membrane protein spectrin Band 2 in Duchenne muscular dystrophy (DMD) was studied in a consecutive series of 10 matched DMD/control pairs. Spectrin was (32P)-phosphorylated by cyclic AMP-independent endogenous membrane protein kinase in the presence of (gamma-32P)ATP. (32P)-labeled spectrin was isolated, purified, and subjected to tryptic cleavage with excess trypsin. The resulting peptides were separated on a high-resolution 5%/15% stacking SDS--polyacrylamide gel electrophoresis system. Liquid scintillation counting was performed on sequential slices of unstained gels. A broad (32P)-labeled band containing a number of (32P)-polypeptides was found to be more highly (32P)-phosphorylated in DMD patients than in their matched controls. This band migrated with an apparent molecular mass of 4.8-5.2 kilodaltons and contained approximately 55% of total (32P) radioactivity covalently bound to spectrin peptides. These data demonstrated an increased (32P)-phosphorylation of an identifiable tryptic peptide fraction in DMD that is consistent with previous reports of increased spectrin Band 2 (32P)-phosphorylation in DMD.

  7. Pepsin and bile acids in saliva in patients with laryngopharyngeal reflux - a prospective comparative study.

    Science.gov (United States)

    Sereg-Bahar, M; Jerin, A; Jansa, R; Stabuc, B; Hocevar-Boltezar, I

    2015-06-01

    Laryngopharyngeal reflux (LPR) and biliary duodenogastric reflux can cause damage to the laryngeal mucosa and voice disorders. The aim of this study was to find out whether levels of pepsin and bile acids in the saliva can serve as diagnostic markers of LPR. A prospective comparative study. Twenty-eight patients with LPR proven via high-resolution manometry and combined multichannel intraluminal impedance and 24-h pH monitoring and 48 healthy controls without symptoms of LPR were included in the study. In the patients with LPR symptoms, oesophagogastroscopy with oesophageal biopsy was performed. The levels of total pepsin, active pepsin, bile acids and the pH of the saliva were determined in all participants and compared between the groups. Reflux symptom index (RSI) and reflux finding score (RFS) were also obtained and compared. The groups differed significantly in RSI (P = 0.00), RFS (P = 0.00), the levels of bile acids (P = 0.005) and total pepsin in saliva (P = 0.023). The levels of total pepsin and bile acids were about three times higher in the patients with LPR than in the healthy controls. There was a significant correlation between the RSI and RFS score and the level of total pepsin and bile acids in the saliva. Histopathological examination of the oesophageal biopsy taken 5 cm above the lower oesophageal sphincter confirmed reflux in almost 93% of patients with symptoms. The study results show that the levels of total pepsin and bile acids in saliva are significantly higher in patients with LPR than in the controls, thus suggesting this as a useful tool in the diagnosis of LPR and particularly biliary LPR. © 2014 John Wiley & Sons Ltd.

  8. Gliadin peptide P31-43 localises to endocytic vesicles and interferes with their maturation.

    Directory of Open Access Journals (Sweden)

    Maria Vittoria Barone

    Full Text Available BACKGROUND: Celiac Disease (CD is both a frequent disease (1:100 and an interesting model of a disease induced by food. It consists in an immunogenic reaction to wheat gluten and glutenins that has been found to arise in a specific genetic background; however, this reaction is still only partially understood. Activation of innate immunity by gliadin peptides is an important component of the early events of the disease. In particular the so-called "toxic" A-gliadin peptide P31-43 induces several pleiotropic effects including Epidermal Growth Factor Receptor (EGFR-dependent actin remodelling and proliferation in cultured cell lines and in enterocytes from CD patients. These effects are mediated by delayed EGFR degradation and prolonged EGFR activation in endocytic vesicles. In the present study we investigated the effects of gliadin peptides on the trafficking and maturation of endocytic vesicles. METHODS/PRINCIPAL FINDINGS: Both P31-43 and the control P57-68 peptide labelled with fluorochromes were found to enter CaCo-2 cells and interact with the endocytic compartment in pulse and chase, time-lapse, experiments. P31-43 was localised to vesicles carrying early endocytic markers at time points when P57-68-carrying vesicles mature into late endosomes. In time-lapse experiments the trafficking of P31-43-labelled vesicles was delayed, regardless of the cargo they were carrying. Furthermore in celiac enterocytes, from cultured duodenal biopsies, P31-43 trafficking is delayed in early endocytic vesicles. A sequence similarity search revealed that P31-43 is strikingly similar to Hrs, a key molecule regulating endocytic maturation. A-gliadin peptide P31-43 interfered with Hrs correct localisation to early endosomes as revealed by western blot and immunofluorescence microscopy. CONCLUSIONS: P31-43 and P57-68 enter cells by endocytosis. Only P31-43 localises at the endocytic membranes and delays vesicle trafficking by interfering with Hrs

  9. Fall in C-peptide during first 2 years from diagnosis: evidence of at least two distinct phases from composite Type 1 Diabetes TrialNet data.

    Science.gov (United States)

    Greenbaum, Carla J; Beam, Craig A; Boulware, David; Gitelman, Stephen E; Gottlieb, Peter A; Herold, Kevan C; Lachin, John M; McGee, Paula; Palmer, Jerry P; Pescovitz, Mark D; Krause-Steinrauf, Heidi; Skyler, Jay S; Sosenko, Jay M

    2012-08-01

    Interpretation of clinical trials to alter the decline in β-cell function after diagnosis of type 1 diabetes depends on a robust understanding of the natural history of disease. Combining data from the Type 1 Diabetes TrialNet studies, we describe the natural history of β-cell function from shortly after diagnosis through 2 years post study randomization, assess the degree of variability between patients, and investigate factors that may be related to C-peptide preservation or loss. We found that 93% of individuals have detectable C-peptide 2 years from diagnosis. In 11% of subjects, there was no significant fall from baseline by 2 years. There was a biphasic decline in C-peptide; the C-peptide slope was -0.0245 pmol/mL/month (95% CI -0.0271 to -0.0215) through the first 12 months and -0.0079 (-0.0113 to -0.0050) from 12 to 24 months (P < 0.001). This pattern of fall in C-peptide over time has implications for understanding trial results in which effects of therapy are most pronounced early and raises the possibility that there are time-dependent differences in pathophysiology. The robust data on the C-peptide obtained under clinical trial conditions should be used in planning and interpretation of clinical trials.

  10. Bond strength of self-etch adhesives after saliva contamination at different application steps.

    Science.gov (United States)

    Cobanoglu, N; Unlu, N; Ozer, F F; Blatz, M B

    2013-01-01

    This study evaluated and compared the effect of saliva contamination and possible decontamination methods on bond strengths of two self-etching adhesive systems (Clearfil SE Bond [CSE], Optibond Solo Plus SE [OSE]). Flat occlusal dentin surfaces were created on 180 extracted human molar teeth. The two bonding systems and corresponding composite resins (Clearfil AP-X, Kerr Point 4) were bonded to the dentin under six surface conditions (n=15/group): group 1 (control): primer/bonding/composite; group 2: saliva/drying/primer/bonding/composite; group 3: primer/saliva/rinsing/drying/primer/bonding/composite; group 4: primer/saliva/rinsing/drying/bonding/composite; group 5: primer/bonding (cured)/saliva/rinsing/drying/primer/bonding/composite; group 6: primer/bonding (cured)/saliva/removing contaminated layer with a bur/rinsing/drying/primer/bonding/composite. Shear bond strength was tested after specimens were stored in distilled water at 37°C for 24 hours. One-way analysis of variance and Tukey post hoc tests were used for statistical analyses. For CSE, groups 2, 3, and 4 and for OSE, groups 6, 2, and 4 showed significantly lower bond strengths than the control group (pcontamination occurred after light polymerization of the bonding agent, repeating the bonding procedure recovered the bonding capacity of both self-etch adhesives. However, saliva contamination before or after primer application negatively affected their bond strength.

  11. Peptide immunisation of HLA-DR-transgenic mice permits the identification of a novel HLA-DRbeta1*0101- and HLA-DRbeta1*0401-restricted epitope from p53.

    Science.gov (United States)

    Rojas, José Manuel; McArdle, Stephanie E B; Horton, Roger B V; Bell, Matthew; Mian, Shahid; Li, Geng; Ali, Selman A; Rees, Robert C

    2005-03-01

    Because of the central role of CD4(+) T cells in antitumour immunity, the identification of the MHC class II-restricted peptides to which CD4(+) T cells respond has become a priority of tumour immunologists. Here, we describe a strategy permitting us to rapidly determine the immunogenicity of candidate HLA-DR-restricted peptides using peptide immunisation of HLA-DR-transgenic mice, followed by assessment of the response in vitro. This strategy was successfully applied to the reported haemaglutinin influenza peptide HA(307-319), and then extended to three candidate HLA-DR-restricted p53 peptides predicted by the evidence-based algorithm SYFPEITHI to bind to HLA-DRbeta1*0101 (HLA-DR1) and HLA-DRbeta1*0401 (HLA-DR4) molecules. One of these peptides, p53(108-122), consistently induced responses in HLA-DR1- and in HLA-DR4-transgenic mice. Moreover, this peptide was naturally processed by dendritic cells (DCs), and induced specific proliferation in the splenocytes of mice immunised with p53 cDNA, demonstrating that immune responses could be naturally mounted to the peptide. Furthermore, p53(108-122) peptide was also immunogenic in HLA-DR1 and HLA-DR4 healthy donors. Thus, the use of this transgenic model permitted the identification of a novel HLA-DR-restricted epitope from p53 and constitutes an attractive approach for the rapid identification of novel immunogenic MHC class II-restricted peptides from tumour antigens, which can ultimately be incorporated in immunotherapeutic protocols.

  12. Associations between food consumption patterns and saliva composition: Specificities of eating difficulties children.

    Science.gov (United States)

    Morzel, Martine; Truntzer, Caroline; Neyraud, Eric; Brignot, Hélène; Ducoroy, Patrick; Lucchi, Géraldine; Canlet, Cécile; Gaillard, Ségolène; Nicod, Florian; Nicklaus, Sophie; Peretti, Noël; Feron, Gilles

    2017-05-01

    Identifying objective markers of diet would be beneficial to research fields such as nutritional epidemiology. As a preliminary study on the validity of using saliva for this purpose, and in order to explore the relationship between saliva and diet, we focused on clearly contrasted groups of children: children with eating difficulties (ED) receiving at least 50% of their energy intake through artificial nutrition vs healthy controls (C). Saliva of ED and C children was analyzed by various methods (targeted biochemical analyses, 2-D electrophoresis coupled to MS, 1 H NMR) and their diet was characterized using food frequency questionnaires, considering 148 food items grouped into 13 categories. Complete datasets were obtained for 16 ED and 16 C subjects (median age 4.7y and 5.0y, respectively) and the statistical link between salivary and dietary characteristics was studied by Multiple Factor Analysis (MFA). Overall, ED children showed as expected lower consumption frequency scores and higher food selectivity. The two groups of children differed in "diet/saliva" associations. Some distinctive salivary variables were common to both groups of children. For example, carbonic anhydrase 6 and the consumption frequency of biscuits & sweets and drinks were positively associated with the MFA axis 1 in C children, but oppositely associated in ED children. Specifically for ED children, abundant salivary proteins (cystatins, amylase, amylase fragments) and some metabolites (amino acids, galactose, lactate) correlated with axis 1, together with the consumption frequency of sauces & seasonings, bread & cereal products, ready-to-eat meals, fish, biscuits & sweets, drinks and potatoes. Specifically for C children, several proteins (serum albumin, haptoglobin, Igκ, apolipoprotein A-1, α-1 antitrypsin) correlated with axis 1, together with the consumption frequency of biscuits & sweets, milk & dairy products, drinks, fruit, meat and vegetables. This study demonstrates that the

  13. Whole Saliva has a Dual Role on the Adherence of Candida albicans to Polymethylmetacrylate

    OpenAIRE

    Elguezabal, N; Maza, J.L.; Dorronsoro, S.; Pont?n, J.

    2008-01-01

    Adhesion of Candida albicans to acrylic of dental prostheses or to salivary macromolecules adsorbed on their surface is believed to be a critical event in the development of denture stomatitis. In previous studies our group has shown that adhesion of C. albicans germ tubes to polystyrene is decreased by saliva whereas C. albicans yeast cells adhesion to the same material is enhanced. The results presented in this study confirm this dual role played by whole saliva, since it decreased the adhe...

  14. The C-Terminal O-S Acyl Shift Pathway under Acidic Condition to Propose Peptide-Thioesters

    Directory of Open Access Journals (Sweden)

    Bo Mi Kim

    2016-11-01

    Full Text Available Peptide-thioester is a pivotal intermediate for peptide ligation and N-, C-terminal cyclization. In this study, desired pathway and the side products of two C-terminal handles, hydroxyethylthiol (HET and hydroxypropylthiol (HPT are described in different conditions as well as kinetic studies. In addition, a new mechanism of C-terminal residue racemization is proposed on the basis of differentiation of products derived from the two C-terminal handles in preparing peptide thioesters through an acid-catalyzed tandem thiol switch, first by an intramolecular O-S acyl shift, and then by an intermolecular S-S exchange.

  15. Measurement of recent exposure to Phlebotomus argentipes, the vector of Indian visceral Leishmaniasis, by using human antibody responses to sand fly saliva.

    Science.gov (United States)

    Clements, Meredith F; Gidwani, Kamlesh; Kumar, Rajiv; Hostomska, Jitka; Dinesh, Diwakar S; Kumar, Vijay; Das, Pradeep; Müller, Ingrid; Hamilton, Gordon; Volfova, Vera; Boelaert, Marleen; Das, Murari; Rijal, Suman; Picado, Albert; Volf, Petr; Sundar, Shyam; Davies, Clive R; Rogers, Matthew E

    2010-05-01

    Antibody (IgG) responses to the saliva of Phlebotomus argentipes were investigated using serum samples from regions of India endemic and non-endemic for visceral leishmaniasis (VL). By pre-adsorbing the sera against the saliva of the competing human-biting but non-VL vector P. papatasi, we significantly improved the specificity of a P. argentipes saliva enzyme-linked immunosorbent assay. Using this method, we observed a statistically significant correlation between antibodies to P. argenitpes saliva and the average indoor density of female sand flies. Additionally, the method was able to detect recent changes in vector exposure when sera from VL patients were assayed before, during, and after hospitalization and protected from sand fly bites under untreated bed nets. Collectively, these results highlight the utility of antibodies to P. argentipes saliva as an important tool to evaluate VL vector control programs.

  16. Contribution of targeted saliva screening for congenital CMV-related hearing loss in newborns who fail hearing screening.

    Science.gov (United States)

    Ari-Even Roth, Daphne; Lubin, Daniel; Kuint, Jacob; Teperberg-Oikawa, Michal; Mendelson, Ella; Strauss, Tzipora; Barkai, Galia

    2017-11-01

    We previously reported a 2.2% rate of infants born with sensorineural hearing loss (SNHL) due to congenital cytomegalovirus (cCMV) infection identified by universal neonatal screen for cCMV using saliva. To evaluate the contribution of targeted saliva screening for cCMV to the detection of infants born with cCMV-related SNHL who failed universal newborn hearing screening (UNHS). We retrospectively reviewed the audiological and medical records of infants who failed UNHS and were tested for cCMV using saliva sample prior to discharge at Sheba Medical Center between 2014 and 2015. Positive cases were confirmed by urine sample. Two hundred (1%) of the 19 830 infants tested during the study period failed in-hospital hearing screening. A saliva specimen was obtained prior to discharge in 187 infants (93.5% of those who failed UNHS). In 178 infants saliva testing was performed at ≤21 days of chronological age and yielded results. cCMV infection was identified in 4/178 tested infants (2.25%, 95% CI 0.8% to 5.3%), of whom three were diagnosed with SNHL (1.7%, 95% CI 0.5% to 4.4%) and offered antiviral treatment. Two of the tested infants (1.12%, 95% CI 0.2% to 3.6%) were diagnosed with cCMV solely due to failure in UNHS. Occult central nervous system (CNS) symptoms of cCMV infection were detected in 2/4 infants following targeted investigation. Targeted cCMV screening in newborns who failed UNHS contributed to the early detection of infants born with cCMV-related isolated SNHL or with occult CNS symptoms who could potentially benefit from antiviral treatment. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  17. White Light Generation in Human Saliva

    Science.gov (United States)

    Santhosh, C.; Dharmadhikari, A. K.; Dharmadhikari, J. A.; Alti, K.; Mathur, D.

    2011-07-01

    Interaction of intense, femto-second pulses of infrared light (800 nm) with water generates white light supercontinuum due to nonlinear optical effects. This supercontinuum was found to be suppressed by the addition of alpha amylase, a major protein in the human saliva. We have studied the suppression of supper continuum by human saliva, collected from healthy subjects with and without smoking habits. Suppression of the blue-sided components was observed significantly in non-smokers saliva than chain smokers.

  18. Innate Immunity and Saliva in Candida albicans–mediated Oral Diseases

    Science.gov (United States)

    Salvatori, O.; Puri, S.; Tati, S.; Edgerton, M.

    2016-01-01

    The oral cavity is a unique niche where Candida albicans infections occur in immunocompetent as well as immunosuppressed individuals. Here we critically review the significance of human innate immune response in preventing oral candidiasis. One important line of defense against oropharyngeal candidiasis is the oral microbiota that prevents infection by competing for space and nutrients as well as by secreting antagonistic molecules and triggering local inflammatory responses. C. albicans is able to induce mucosal defenses through activation of immune cells and production of cytokines. Also, saliva contains various proteins that affect C. albicans growth positively by promoting mucosal adherence and negatively through immune exclusion and direct fungicidal activity. We further discuss the role of saliva in unifying host innate immune defenses against C. albicans as a communicating medium and how C. albicans overgrowth in the oral cavity may be a result of aberrations ranging from microbial dysbiosis and salivary dysfunction to epithelial damage. Last we underscore select oral diseases in which C. albicans is a contributory microorganism in immune-competent individuals. PMID:26747422

  19. Changes in the Concentration of Ions in Saliva and Dental Plaque after Application of CPP-ACP with and without Fluoride among 6-9 Year Old Children

    Directory of Open Access Journals (Sweden)

    Poureslami H

    2017-03-01

    Full Text Available Statement of Problem: The casein phospho peptide-amorphous calcium phosphate with or without fluoride (CPP-ACPF and CPP-ACP respectively are of considerably new materials which are highly recommended for prevention of dental caries. However, there is a shortage in literature on how they affect the ion concentration of saliva or dental plaque. Objectives: The aim of this study was to evaluate the concentration of calcium, phosphate and fluoride in the plaque and saliva of children with Early Childhood Caries (ECC after applying the CPP-ACP paste in comparison with the use of CPP- ACPF paste. Materials and Methods: One ml of un-stimulated saliva of 25 preschool children was collected and then 1 mg of the plaque sample was collected from the buccal surfaces of the two first primary molars on the upper jaw. CPP-ACP as well as CPP- ACPF pastes were applied on the tooth surfaces in two separate steps. In steps, plaque and saliva sampling was performed after 60 minutes. The amount of calcium ions was measured by Atomic Absorption Device and the amount of phosphate and fluoride ions was measured by Ion Chromatography instrument. Data were analyzed using Repeated Measurements ANOVA at a p < 0.05 level of significance. Results: Application of both CPP-ACPF and CPP-ACP significantly increased the concentration of calcium, phosphate, and fluoride in both saliva and dental plaque. Moreover, significantly higher salivary fluoride concentration was seen after application of CPP-ACPF compared to CPP-ACP. No other significant difference was observed between these two materials. Conclusions: CPP-ACPF can be more useful than CPP-ACP in protecting the primary teeth against caries process, especially when there is poor hygiene.

  20. Rhizobial peptidase HrrP cleaves host-encoded signaling peptides and mediates symbiotic compatibility.

    Science.gov (United States)

    Price, Paul A; Tanner, Houston R; Dillon, Brett A; Shabab, Mohammed; Walker, Graham C; Griffitts, Joel S

    2015-12-08

    Legume-rhizobium pairs are often observed that produce symbiotic root nodules but fail to fix nitrogen. Using the Sinorhizobium meliloti and Medicago truncatula symbiotic system, we previously described several naturally occurring accessory plasmids capable of disrupting the late stages of nodule development while enhancing bacterial proliferation within the nodule. We report here that host range restriction peptidase (hrrP), a gene found on one of these plasmids, is capable of conferring both these properties. hrrP encodes an M16A family metallopeptidase whose catalytic activity is required for these symbiotic effects. The ability of hrrP to suppress nitrogen fixation is conditioned upon the genotypes of both the host plant and the hrrP-expressing rhizobial strain, suggesting its involvement in symbiotic communication. Purified HrrP protein is capable of degrading a range of nodule-specific cysteine-rich (NCR) peptides encoded by M. truncatula. NCR peptides are crucial signals used by M. truncatula for inducing and maintaining rhizobial differentiation within nodules, as demonstrated in the accompanying article [Horváth B, et al. (2015) Proc Natl Acad Sci USA, 10.1073/pnas.1500777112]. The expression pattern of hrrP and its effects on rhizobial morphology are consistent with the NCR peptide cleavage model. This work points to a symbiotic dialogue involving a complex ensemble of host-derived signaling peptides and bacterial modifier enzymes capable of adjusting signal strength, sometimes with exploitative outcomes.

  1. Solubility of root-canal sealers in water and artificial saliva.

    Science.gov (United States)

    Schäfer, E; Zandbiglari, T

    2003-10-01

    To compare the weight loss of eight different root-canal sealers in water and in artificial saliva with different pH values. For standardized samples (n = 12 per group), ring moulds were filled with epoxy resin (AH 26, AH Plus)-, silicone (RSA RoekoSeal)-, calcium hydroxide (Apexit, Sealapex)-, zinc oxide-eugenol (Aptal-Harz)-, glass-ionomer (Ketac Endo)- and polyketone (Diaket)-based sealers. These samples were immersed in double-distilled water or artificial saliva with different pH values (7.0, 5.7 and 4.5) for 30 s, 1 min, 2 min, 5 min, 10 min, 20 min, 1 h, 2 h, 10 h, 24 h, 48 h, 72 h, 14 days and 28 days. Mean loss of weight was determined and analysed statistically using a one-way anova and Student-Newman-Keuls test for all pairwise comparisons. Most sealers were of low solubility, although Sealapex, Aptal-Harz and Ketac Endo showed a marked weight loss in all liquids. Even after 28 days of storage in water, AH 26, AH Plus, RSA RoekoSeal, and Diaket showed less than 3% weight loss. At exposure times greater than 14 days, Sealapex showed the significantly greatest weight loss of all sealers tested (P < 0.05). Aptal-Harz and Ketac Endo were significantly more soluble in saliva (pH 4.5) than in water (P < 0.05). Under the conditions of the present study, AH Plus showed the least weight loss of all sealers tested, independent of the solubility medium used. Sealapex, Aptal-Harz and Ketac Endo had a marked weight loss in all liquids.

  2. Sensitivity and reproducibility of urinary C-peptide as estimate of islet B-cell function in insulin-treated diabetes

    DEFF Research Database (Denmark)

    Gjessing, H J; Matzen, L E; Faber, O K

    1989-01-01

    The aims of the present study were to evaluate the ability of urinary C-peptide determination to demonstrate presence of residual insulin secretion, and to evaluate the reproducibility of urinary C-peptide excretion in 125 insulin-treated diabetic patients. C-peptide was determined in two...

  3. Porphyromonas pasteri sp. nov., isolated from human saliva.

    Science.gov (United States)

    Sakamoto, Mitsuo; Li, Dan; Shibata, Yukie; Takeshita, Toru; Yamashita, Yoshihisa; Ohkuma, Moriya

    2015-08-01

    A bacterial strain, designated KUFDS01T, isolated from human saliva was characterized using a polyphasic taxonomic approach that included analysis of physiological and biochemical features, cellular fatty acid profiles and phylogenetic position based on 16S rRNA gene sequence analysis. Cells of the strain were obligately anaerobic, non-pigmented, non-spore-forming, non-motile, Gram-stain-negative rods. Growth of the strain was inhibited on medium containing 20% bile. The 16S rRNA gene sequence analysis showed that the strain was a member of the genus Porphyromonas. Strain KUFDS01T was closely related to Porphyromonas catoniae JCM 13863T (96.6% sequence similarity). An hsp60 gene sequence analysis indicated that strain KUFDS01T was different from P. catoniae JCM 13863T, with a sequence similarity value of 87.8%. The major cellular fatty acids of strain KUFDS01T were C16 : 0, iso-C15 : 0, anteiso-C15 : 0, C18 : 2ω6, 9c and C18 : 1ω9c. The DNA G+C content of strain KUFDS01T was 57.7 ± 0.66 mol%. On the basis of these data, strain KUFDS01T represents a novel species of the genus Porphyromonas, for which the name Porphyromonas pasteri sp. nov. is proposed. The type strain of P. pasteri is KUFDS01T ( = JCM 30531T = CCUG 66735T).

  4. Fasting serum C-peptide is useful for initial classification of diabetes mellitus in children and adolescents

    Directory of Open Access Journals (Sweden)

    Min Jung Cho

    2014-06-01

    Full Text Available PurposeWith rising obesity rates in children, it is increasingly difficult to differentiate between type 1 and type 2 diabetes mellitus (T1DM, T2DM on clinical grounds alone. Using C-peptide as a method of classifying diabetes mellitus (DM has been suggested. This study aimed to find a correlation between fasting C-peptide level and DM types in children and adolescents.MethodsA total of 223 diabetic children, newly diagnosed at 5 hospitals between January 2001 and December 2012, were enrolled in this study. Initial DM classification was based on clinical and laboratory data including fasting C-peptide at diagnosis; final classification was based on additional data (pancreatic autoantibodies, human leukocyte antigen type, and clinical course.ResultsOf 223 diabetic children, 140 were diagnosed with T1DM (62.8% and the remaining 83 with T2DM (37.2%. The mean serum C-peptide level was significantly lower in children with T1DM (0.80 ng/mL than in children with T2DM (3.91 ng/mL. Among 223 children, 54 had a serum C-peptide level 3.0 ng/mL; 48 of them (97.9% were diagnosed with T2DM.ConclusionIn this study, we found that if the C-peptide level was 3.0 ng/mL, a T1DM diagnosis is unlikely. This finding suggests that serum fasting C-peptide level is useful for classifying DM type at the time of diagnosis in youth.

  5. Peptide p5 binds both heparinase-sensitive glycosaminoglycans and fibrils in patient-derived AL amyloid extracts

    Energy Technology Data Exchange (ETDEWEB)

    Martin, Emily B.; Williams, Angela [Department of Medicine, University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, TN 37922 (United States); Heidel, Eric [Department of Surgery, University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, TN 37922 (United States); Macy, Sallie [Department of Medicine, University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, TN 37922 (United States); Kennel, Stephen J. [Department of Medicine, University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, TN 37922 (United States); Department of Radiology, University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, TN 37922 (United States); Wall, Jonathan S., E-mail: jwall@utmck.edu [Department of Medicine, University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, TN 37922 (United States); Department of Radiology, University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, TN 37922 (United States)

    2013-06-21

    Highlights: •Polybasic peptide p5 binds human light chain amyloid extracts. •The binding of p5 with amyloid involves both glycosaminoglycans and fibrils. •Heparinase treatment led to a correlation between p5 binding and fibril content. •p5 binding to AL amyloid requires electrostatic interactions. -- Abstract: In previously published work, we have described heparin-binding synthetic peptides that preferentially recognize amyloid deposits in a mouse model of reactive systemic (AA) amyloidosis and can be imaged by using positron and single photon emission tomographic imaging. We wanted to extend these findings to the most common form of visceral amyloidosis, namely light chain (AL); however, there are no robust experimental animal models of AL amyloidosis. To further define the binding of the lead peptide, p5, to AL amyloid, we characterized the reactivity in vitro of p5 with in situ and patient-derived AL amyloid extracts which contain both hypersulfated heparan sulfate proteoglycans as well as amyloid fibrils. Histochemical staining demonstrated that the peptide specifically localized with tissue-associated AL amyloid deposits. Although we anticipated that p5 would undergo electrostatic interactions with the amyloid-associated glycosaminoglycans expressing heparin-like side chains, no significant correlation between peptide binding and glycosaminoglycan content within amyloid extracts was observed. In contrast, following heparinase I treatment, although overall binding was reduced, a positive correlation between peptide binding and amyloid fibril content became evident. This interaction was further confirmed using synthetic light chain fibrils that contain no carbohydrates. These data suggest that p5 can bind to both the sulfated glycosaminoglycans and protein fibril components of AL amyloid. Understanding these complex electrostatic interactions will aid in the optimization of synthetic peptides for use as amyloid imaging agents and potentially as

  6. Radioimmunoassay of seric C-peptide. Practical value in the study of insulin secretion. Results of 140 stimulation tests

    International Nuclear Information System (INIS)

    Wafflart, Jean.

    1977-10-01

    C-peptide, which appears as a by-product of insulin synthesis, is secreted with this latter in equimolar quantities but is not degraded in the liver. It thus reflects indirectly the insulin secreted. After the structure of C-peptide was determined in 1971 by OYER it was synthesized by YANAIHARA and a radioimmunoassay was developed by KANEKO in 1974. This work was made possible by the recent commercialisation of a Japanese analysis kit, the 'DAIICHI' kit, and its availability through GUERBET TESTS. Part one describes the structural, physiological and immuno properties of C-peptide and its method of determination. Part two is devoted to a review of foreign publications on the practical interest of the C-peptide measurement. Part three gives the results of 140 oral or venous stimulation tests where blood sugar, blood insulin and C-peptide are measured in parallel. The different diabetic pathologies are explored and compared against normal subjects. The purpose of this work is to establish the value of C-peptide as a reflection of insulin secretion on the one hand, and that of a parallel insulin and C-peptide determination on the other [fr

  7. Optimization of reagent concentration for radioiodination of rat C-peptide II in development of radioimmunoassay procedure for rats

    Directory of Open Access Journals (Sweden)

    B R Manupriya

    2018-01-01

    Full Text Available Rat C-peptide is a polypeptide molecule made up of 31 amino acids and secreted from pancreas into circulation in two isoforms I and II. Quantification of rat C-peptide II in rat serum is important as it is directly related to the diagnosis of carbohydrate metabolism abnormalities, pancreatic performance analysis, monitoring of hypoglycemia, and diabetes-related illness in rat model. The aim of the present work is to develop a tracer by chloramine-T method for radioimmunoassay (RIA procedure and to determine the optimum amount of chloramine-T required for the preparation of stable radioiodinated product with a specific activity of around 24.97 MBq/μg, corresponding to 1 125I atom per molecule of the peptide. Tyrosylated rat C-peptide II was selected for the radioiodination procedure as rat C-peptide II does not contain either tyrosine or histidine which is mandatory for the incorporation of 125I atom to the rat C-peptide II. Tyrosylated rat C-peptide II was subjected to radioiodination by chloramine-T method with different concentrations of chloramine-T and sodium metabisulfite (MBS to obtain a stable radiolabeled compound. Optimized reaction conditions relating to the concentration of chloramine-T (10 μg and MBS (20 μg yielded a stable 125I-rat C-peptide II with specific activity of 21.01 MBq/μg corresponding to 0.84 125I atoms per molecule of the peptide. Preparation of high integrity tracer of rat C-peptide II was achieved by combining one molecule of oxidant (chloramine-T and two molecule of reductant (MBS.

  8. Correlation of Dental Caries Experience with pH, Buffering Capacity and Flow Rate of Saliva among 15-year-old School Children in Mangalore

    Directory of Open Access Journals (Sweden)

    Vijay S Kumar

    2013-01-01

    Full Text Available Aim and Objectives: To correlate dental caries experience with pH, buffering capacity and flow rate of saliva among 15-year-old-children Materials & Method : Fifteen year old school children from adopted schools of a dental college in Mangalore were considered as study subjects. The study was carried out using a standardized proforma that consisted of questionnaire for recording demographic data. The caries experience was assessed by DMFT Index. Salivary pH was measured by using a pH meter and salivary buffering capacity was measured by using titration method. Unstimulated whole saliva was collected from all subjects by direct expectoration to calculate the flow rate. Results: The mean DMFT of the study population was 2.85 ± 2.5. The mean salivary pH was found to be 6.88 ± 0.69.About 74.1% were having medium salivary buffering capacities. The mean unstimulated salivary flow rate and mean total antioxidant capacity of the study population was 0.41 ± 0.14 ml/min and 94.15 ± 60.72 g/dL respectively. Conclusions: A marked association between the pH, buffering capacity and unstimulated flow rate with dental caries experience suggest that assessment of these salivary parameters can be used as predictors for future dental caries susceptibility in an individual.

  9. Elevated C-peptide and insulin predict increased risk of colorectal adenomas in normal mucosa

    International Nuclear Information System (INIS)

    Vidal, Adriana C; Keku, Temitope O; Lund, Pauline Kay; Hoyo, Cathrine; Galanko, Joseph; Burcal, Lauren; Holston, Rachel; Massa, Berri; Omofoye, Oluwaseun; Sandler, Robert S

    2012-01-01

    Lower concentrations of the insulin–like growth factor binding protein-1 (IGFBP-1) and elevated concentrations of insulin or C-peptide have been associated with an increase in colorectal cancer risk (CRC). However few studies have evaluated IGFBP-1 and C-peptide in relation to adenomatous polyps, the only known precursor for CRC. Between November 2001 and December 2002, we examined associations between circulating concentrations of insulin, C-peptide, IGFBP-1 and apoptosis among 190 individuals with one or more adenomatous polyps and 488 with no adenomatous polyps using logistic regression models. Individuals with the highest concentrations of C-peptide were more likely to have adenomas (OR = 2.2, 95% CI 1.4-4.0) than those with the lowest concentrations; associations that appeared to be stronger in men (OR = 4.4, 95% CI 1.7-10.9) than women. Individuals with high insulin concentrations also had a higher risk of adenomas (OR = 3.5, 95% CI 1.7-7.4), whereas higher levels of IGFBP-1 were associated with a reduced risk of adenomas in men only (OR = 0.3, 95% CI 0.1-0.7). Overweight and obese individuals with higher C-peptide levels (>1 st Q) were at increased risk for lower apoptosis index (OR = 2.5, 95% CI 0.9-7.1), an association that remained strong in overweight and obese men (OR = 6.3, 95% CI 1.0-36.7). Higher levels of IGFBP-1 in overweight and obese individuals were associated with a reduced risk of low apoptosis (OR = 0.3, 95% CI 0.1-1.0). Associations between these peptides and the apoptosis index in overweight and obese individuals, suggest that the mechanism by which C-peptide could induce adenomas may include its anti-apoptotic properties. This study suggests that hyperinsulinemia and IGF hormones predict adenoma risk, and that outcomes associated with colorectal carcinogenesis maybe modified by gender

  10. Effect of endurance training on dental erosion, caries, and saliva.

    Science.gov (United States)

    Frese, C; Frese, F; Kuhlmann, S; Saure, D; Reljic, D; Staehle, H J; Wolff, D

    2015-06-01

    The aim of this investigation was to give insights into the impact of endurance training on oral health, with regard to tooth erosion, caries, and salivary parameters. The study included 35 triathletes and 35 non-exercising controls. The clinical investigation comprised oral examination, assessment of oral status with special regard to caries and erosion, saliva testing during inactivity, and a self-administered questionnaire about eating, drinking, and oral hygiene behavior. In addition, athletes were asked about their training habits and intake of beverages and sports nutrition. For saliva assessment during exercise, a subsample of n = 15 athletes volunteered in an incremental running field test (IRFT). Athletes showed an increased risk for dental erosion (P = 0.001). No differences were observed with regard to caries prevalence and salivary parameters measured during inactivity between athletes and controls. Among athletes, a significant correlation was found between caries prevalence and the cumulative weekly training time (r = 0.347, P = 0.04). In athletes after IRFT and at maximum workload, saliva flow rates decreased (P = 0.001 stimulated; P = 0.01 unstimulated) and saliva pH increased significantly (P = 0.003). Higher risk for dental erosions, exercise-dependent caries risk, and load-dependent changes in saliva parameters point out the need for risk-adapted preventive dental concepts in the field of sports dentistry. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  11. Discovery of Novel Antimicrobial Peptides from Varanus komodoensis (Komodo Dragon) by Large-Scale Analyses and De-Novo-Assisted Sequencing Using Electron-Transfer Dissociation Mass Spectrometry.

    Science.gov (United States)

    Bishop, Barney M; Juba, Melanie L; Russo, Paul S; Devine, Megan; Barksdale, Stephanie M; Scott, Shaylyn; Settlage, Robert; Michalak, Pawel; Gupta, Kajal; Vliet, Kent; Schnur, Joel M; van Hoek, Monique L

    2017-04-07

    Komodo dragons are the largest living lizards and are the apex predators in their environs. They endure numerous strains of pathogenic bacteria in their saliva and recover from wounds inflicted by other dragons, reflecting the inherent robustness of their innate immune defense. We have employed a custom bioprospecting approach combining partial de novo peptide sequencing with transcriptome assembly to identify cationic antimicrobial peptides from Komodo dragon plasma. Through these analyses, we identified 48 novel potential cationic antimicrobial peptides. All but one of the identified peptides were derived from histone proteins. The antimicrobial effectiveness of eight of these peptides was evaluated against Pseudomonas aeruginosa (ATCC 9027) and Staphylococcus aureus (ATCC 25923), with seven peptides exhibiting antimicrobial activity against both microbes and one only showing significant potency against P. aeruginosa. This study demonstrates the power and promise of our bioprospecting approach to cationic antimicrobial peptide discovery, and it reveals the presence of a plethora of novel histone-derived antimicrobial peptides in the plasma of the Komodo dragon. These findings may have broader implications regarding the role that intact histones and histone-derived peptides play in defending the host from infection. Data are available via ProteomeXChange with identifier PXD005043.

  12. A Maple Program That Illustrates the Effect of pH on Peptide Charge

    Science.gov (United States)

    Sokolik, Charles W.

    1998-11-01

    One topic covered early in an introductory biochemistry course is acid-base chemistry and the Henderson-Hasselbalch equation (buffer equation). Using this equation a biochemistry student can determine the partial charges of amino acids in a peptide chain. This is an important concept to master for a student who is learning the structure-function relationship in proteins. The program described in this paper, written for Maple V, release 3 (Waterloo Maple Software, Waterloo, ON, Canada), uses the Henderson-Hasselbalch equation to calculate the partial charges of individual amino acids and the net charge of a peptide over the pH range 0 to 14. The amino acid sequence of a peptide is entered and an animated histogram is displayed illustrating the partial charge of the amino acids over the pH range. A graph showing the net charge of the peptide from pH 0 to 14 is also given. The program has been used with success in an introductory biochemistry course as an in-class demonstration as well as for individual homework assignments. The program is available through the Web page of the Journal of Chemical Education.

  13. Morphology and Differentiation of MG63 Osteoblast Cells on Saliva Contaminated Implant Surfaces

    Directory of Open Access Journals (Sweden)

    Neda Shams

    2015-11-01

    Full Text Available Objectives: Osteoblasts are the most important cells in the osseointegration process. Despite years of study on dental Implants, limited studies have discussed the effect of saliva on the adhesion process of osteoblasts to implant surfaces. The aim of this in vitro study was to evaluate the effect of saliva on morphology and differentiation of osteoblasts attached to implant surfaces.Materials and Methods: Twelve Axiom dental implants were divided into two groups. Implants of the case group were placed in containers, containing saliva, for 40 minutes. Then, all the implants were separately stored in a medium containing MG63 human osteoblasts for a week. Cell morphology and differentiation were assessed using a scanning electron microscope and their alkaline phosphatase (ALP activity was determined. The t-test was used to compare the two groups.Results: Scanning electron microscopic observation of osteoblasts revealed round or square cells with fewer and shorter cellular processes in saliva contaminated samples, whereas elongated, fusiform and well-defined cell processes were seen in the control group. ALP level was significantly lower in case compared to control group (P<0.05.Conclusion: Saliva contamination alters osteoblast morphology and differentiation and may subsequently interfere with successful osseointegration. Thus, saliva contamination of bone and implant must be prevented or minimized.

  14. C-type natriuretic peptide in prostate cancer

    DEFF Research Database (Denmark)

    Nielsen, Soeren Junge; Iversen, Peter; Rehfeld, Jens F.

    2009-01-01

    C-type natriuretic peptide (CNP) is expressed in the male reproductive organs in pigs. To examine whether the human prostate also expresses the CNP gene, we measured CNP and N-terminal proCNP in prostate cancer tissue extracts and performed immunohistochemical biopsy staining. Additionally, pro......CNP-derived peptides were quantitated in plasma from patients with prostate cancer. Blood was collected from healthy controls and patients before surgery for localized prostate cancer. Tissue extracts were prepared from tissue biopsies obtained from radical prostatectomy surgery. N-terminal proCNP, proCNP (1......-50) and CNP were measured in plasma and tissue extracts. Biopsies were stained for CNP-22 and N-terminal proCNP. Tissue extracts from human prostate cancer contained mostly N-terminal proCNP [median 5.3 pmol/g tissue (range 1.0-12.9)] and less CNP [0.14 pmol/g tissue (0.01-1.34)]. Immunohistochemistry...

  15. The functions of human saliva

    DEFF Research Database (Denmark)

    Dawes, C; Pedersen, Anne Marie Lynge; Villa, A

    2015-01-01

    This narrative review of the functions of saliva was conducted in the PubMed, Embase and Web of Science databases. Additional references relevant to the topic were used, as our key words did not generate references which covered all known functions of saliva. These functions include maintaining a...... of oral wounds. Clearly, saliva has many functions which are needed for proper protection and functioning of the human body....... a moist oral mucosa which is less susceptible to abrasion, and removal of micro-organisms, desquamated epithelial cells, leucocytes and food debris by swallowing. The mucins form a slimy coating on all surfaces in the mouth and act as a lubricant during such processes as mastication, formation of a food...

  16. The clinical value of detecting INS and C - P in essential hypertension

    International Nuclear Information System (INIS)

    Huang Jianmin; Sun Shuqiang; Liu Xiaomei

    2001-01-01

    In order to explore the clinical value of insulin (INS) and C - peptide (C-P) in essential hypertension patients (EH), concentrations of blood glucose, serum INS and serum C - P both during fasting and 2 - hour postprandial time were measured in 32 non-diabetic patients with EH and 40 healthy persons, and the ISI of each group was calculated and compared. The results showed that with the exception of fasting blood glucose level being of no difference between two groups, all the other parameters observed, including concentrations of 2 hour postprandial glucose, both fasting and 2 hour postprandial INS and C - P were significantly higher than those of healthy control group (P < 0.01). The proportions of patients with insulin resistance and hyperinsulinemia in EH group were both significantly higher than those of control group (P < 0.01). These results suggest that EH patients may coexist with some metabolic disorders at the same time, thus INS and C - P detection may have other wider applications in non-diabetic diseases

  17. Radioimmunological analysis of circadian rhythms of cortisol and melatonin in saliva

    International Nuclear Information System (INIS)

    Demel, A.W.

    1990-12-01

    Since blood cortisol (F) and melatonin (MLT) display a circadian secretion pattern and since the saliva concentration of this hormones is an excellent indicator of its blood levels the measurement of salivary F and MLT may be used for examining circadian rhythmicity. In this study the relationship between salivary F and MLT was explored. For this purpose it was necessary first to establish and validate a radioimmunoassay for F in saliva: salivary F was determined by a direct radioimmunoassay using cortisol-3-(O-carboxymethyl) oximino-(2-( 125 I)iodohistamin) as tracer and cortisol-3-CMO-BSA antiserum. The parallel measurement of F levels in saliva and serum of adults gave an excellent correlation (r=0.87, p 0.00956x ). Serum F was assayed on the Abott TDX-System using a radioimmunofluorescence method. Secondly, using this assay the circadian saliva F pattern was determined as well as the pattern of salivary MLT in 9 young, healthy volunteers. For saliva MLT estimations a previously published method was applied (Schulz et al 1990). Using a computerized program (RHYTHM) written by Eve v. Cauter (1979), the hormone data of each individuum were examined for circadian rhythmicity and its acrophase (time of occurrence of the maximum of a sinusoid fitted to the data). The F acrophase occurred between 7:00 and 12:00 h (mean: 3:33 h, SD: 104.4 min). The easy stress-free non invasive nature of saliva collection makes saliva to one of the most accessible body fluids and of high value in studying the circadian system in healthy humans as well as in infants, children, pregnant women and anaemic patients. Measurements of salivary F and MLT may help to elucidate not only the circadian rhythms of these hormones under normal and pathological conditions but it may also provide insight in physiology and pathology of the circadian system in general. (author)

  18. BnaA.bZIP1 Negatively Regulates a Novel Small Peptide Gene, BnaC.SP6, Involved in Pollen Activity

    Directory of Open Access Journals (Sweden)

    Xuanpeng Wang

    2017-12-01

    Full Text Available Small peptides secreted to the extracellular matrix control many aspects of the plant’s physiological activities which were identified in Arabidopsis thaliana, called ATSPs. Here, we isolated and characterized the small peptide gene Bna.SP6 from Brassica napus. The BnaC.SP6 promoter was cloned and identified. Promoter deletion analysis suggested that the -447 to -375 and -210 to -135 regions are crucial for the silique septum and pollen expression of BnaC.SP6, respectively. Furthermore, the minimal promoter region of p158 (-210 to -52 was sufficient for driving gene expression specifically in pollen and highly conserved in Brassica species. In addition, BnaA.bZIP1 was predominantly expressed in anthers where BnaC.SP6 was also expressed, and was localized to the nuclei. BnaA.bZIP1 possessed transcriptional activation activity in yeast and protoplast system. It could specifically bind to the C-box in p158 in vitro, and negatively regulate p158 activity in vivo. BnaA.bZIP1 functions as a transcriptional repressor of BnaC.SP6 in pollen activity. These results provide novel insight into the transcriptional regulation of BnaC.SP6 in pollen activity and the pollen/anther-specific promoter regions of BnaC.SP6 may have their potential agricultural application for new male sterility line generation.

  19. 21 CFR 862.1135 - C-peptides of proinsulin test system.

    Science.gov (United States)

    2010-04-01

    .... Measurements of C-peptides of proinsulin are used in the diagnosis and treatment of patients with abnormal insulin secretion, including diabetes mellitus. (b) Classification. Class I (general controls). The device...

  20. NMR and molecular modeling of wine tannins binding to saliva proteins: revisiting astringency from molecular and colloidal prospects.

    Science.gov (United States)

    Cala, Olivier; Pinaud, Noël; Simon, Cécile; Fouquet, Eric; Laguerre, Michel; Dufourc, Erick J; Pianet, Isabelle

    2010-11-01

    In organoleptic science, the association of tannins to saliva proteins leads to the poorly understood phenomenon of astringency. To decipher this interaction at molecular and colloidal levels, the binding of 4 procyanidin dimers (B1-4) and 1 trimer (C2) to a human saliva proline-rich peptide, IB7(14), was studied. Interactions have been characterized by measuring dissociation constants, sizes of complexes, number, and nature of binding sites using NMR (chemical shift variations, diffusion-ordered spectroscopy, and saturation transfer diffusion). The binding sites were identified using molecular mechanics, and the hydrophilic/hydrophobic nature of the interactions was resolved by calculating the molecular lipophilicity potential within the complexes. The following comprehensive scheme can be proposed: 1) below the tannin critical micelle concentration (CMC), interaction is specific, and the procyanidin anchorage always occurs on the same three IB7(14) sites. The tannin 3-dimensional structure plays a key role in the binding force and in the tannin's ability to act as a bidentate ligand: tannins adopting an extended conformation exhibit higher affinity toward protein and initiate the formation of a network. 2) Above the CMC, after the first specific hydrophilic interaction has taken place, a random hydrophobic stacking occurs between tannins and proteins. The whole process is discussed in the general frame of wine tannins eliciting astringency.

  1. Cortisol in urine and saliva

    DEFF Research Database (Denmark)

    Hurwitz Eller, N; Netterstrøm, B; Hansen, Åse Marie

    2001-01-01

    The objective of the study was to analyse the relations between excretion of cortisol in urine and saliva and the intima media thickness (IMT) of the artery carotis communis.......The objective of the study was to analyse the relations between excretion of cortisol in urine and saliva and the intima media thickness (IMT) of the artery carotis communis....

  2. Computational strategy for quantifying human pesticide exposure based upon a saliva measurement

    Energy Technology Data Exchange (ETDEWEB)

    Timchalk, Charles; Weber, Thomas J.; Smith, Jordan N.

    2015-05-27

    The National Research Council of the National Academies report, Toxicity Testing in the 21st Century: A Vision and Strategy, highlighted the importance of quantitative exposure data for evaluating human toxicity risk and noted that biomonitoring is a critical tool for quantitatively evaluating exposure from both environmental and occupational settings. Direct measurement of chemical exposures using personal monitoring provides the most accurate estimation of a subject’s true exposure, and non-invasive methods have also been advocated for quantifying the pharmacokinetics and bioavailability of drugs and xenobiotics. In this regard, there is a need to identify chemicals that are readily cleared in saliva at concentrations that can be quantified to support the implementation of this approach.. The current manuscript describes the use of computational modeling approaches that are closely coupled to in vivo and in vitro experiments to predict salivary uptake and clearance of xenobiotics. The primary mechanism by which xenobiotics leave the blood and enter saliva is thought to involve paracellular transport, passive transcellular diffusion, or trancellular active transport with the majority of drugs and xenobiotics cleared from plasma into saliva by passive diffusion. The transcellular or paracellular diffusion of unbound chemicals in plasma to saliva has been computational modeled using a combination of compartmental and physiologically based approaches. Of key importance for determining the plasma:saliva partitioning was the utilization of a modified Schmitt algorithm that calculates partitioning based upon the tissue composition, pH, chemical pKa and plasma protein-binding. Sensitivity analysis of key model parameters specifically identified that both protein-binding and pKa (for weak acids and bases) had the most significant impact on the determination of partitioning and that there were clear species dependent differences based upon physiological variance between

  3. Bacterial profiles of saliva in relation to diet, lifestyle factors, and socioeconomic status

    Directory of Open Access Journals (Sweden)

    Daniel Belstrøm

    2014-04-01

    Full Text Available Background and objective: The bacterial profile of saliva is composed of bacteria from different oral surfaces. The objective of this study was to determine whether different diet intake, lifestyle, or socioeconomic status is associated with characteristic bacterial saliva profiles. Design: Stimulated saliva samples from 292 participants with low levels of dental caries and periodontitis, enrolled in the Danish Health Examination Survey (DANHES, were analyzed for the presence of approximately 300 bacterial species by means of the Human Oral Microbe Identification Microarray (HOMIM. Using presence and levels (mean HOMIM-value of bacterial probes as endpoints, the influence of diet intake, lifestyle, and socioeconomic status on the bacterial saliva profile was analyzed by Mann–Whitney tests with Benjamini–Hochberg's correction for multiple comparisons and principal component analysis. Results: Targets for 131 different probes were identified in 292 samples, with Streptococcus and Veillonella being the most predominant genera identified. Two bacterial taxa (Streptococcus sobrinus and Eubacterium [11][G-3] brachy were more associated with smokers than non-smokers (adjusted p-value<0.01. Stratification of the group based on extreme ends of the parameters age, gender, alcohol consumption, body mass index (BMI, and diet intake had no statistical influence on the composition of the bacterial profile of saliva. Conversely, differences in socioeconomic status were reflected by the bacterial profiles of saliva. Conclusions: The bacterial profile of saliva seems independent of diet intake, but influenced by smoking and maybe socioeconomic status.

  4. Peptides-Derived from Thai Rice Bran Improves Endothelial Function in 2K-1C Renovascular Hypertensive Rats

    Directory of Open Access Journals (Sweden)

    Orachorn Boonla

    2015-07-01

    Full Text Available In recent years, a number of studies have investigated complementary medical approaches to the treatment of hypertension using dietary supplements. Rice bran protein hydrolysates extracted from rice is a rich source of bioactive peptides. The present study aimed to investigate the vasorelaxation and antihypertensive effects of peptides-derived from rice bran protein hydrolysates (RBP in a rat model of two kidney-one clip (2K-1C renovascular hypertension. 2K-1C hypertension was induced in male Sprague-Dawley rats by placing a silver clip around the left renal artery, whereas sham-operated rats were served as controls. 2K-1C and sham-operated rats were intragastrically administered with RBP (50 mg kg−1 or 100 mg kg−1 or distilled water continuously for six weeks. We observed that RBP augmented endothelium-dependent vasorelaxation in all animals. Administration of RBP to 2K-1C rats significantly reduced blood pressure and decreased peripheral vascular resistance compared to the sham operated controls (p < 0.05. Restoration of normal endothelial function and blood pressure was associated with reduced plasma angiotensin converting enzyme (ACE, decreased superoxide formation, reduced plasma malondialdehyde and increased plasma nitrate/nitrite (p < 0.05. Up-regulation of eNOS protein and down-regulation of p47phox protein were found in 2K-1C hypertensive rats-treated with RBP. Our results suggest that RBP possesses antihypertensive properties which are mainly due to the inhibition of ACE, and its vasodilatory and antioxidant activity.

  5. Recuperación de veillonellas a partir de saliva Recovery of Veillonella from saliva

    Directory of Open Access Journals (Sweden)

    M.I. Gutiérrez De Ferro

    2005-03-01

    Full Text Available Las veillonellas son cocos gram-negativos anaerobios asociados con salud oral. Para su aislamiento, se han reportado diferentes medios de cultivo. Las colonias de Veillonella spp. producen fluorescencia roja visible con luz ultravioleta, que desaparece en contacto con oxígeno. Esta propiedad sería útil para su identificación presuntiva rápida. Los objetivos de este trabajo fueron: 1- comparar el medio selectivo para Veillonella de Rogosa con los medios de cultivo recomendados por diferentes autores para determinar en cual de ellos se obtiene una mejor recuperación de veillonellas a partir de saliva, ya que esta muestra es generalmente utilizada para determinar la presencia y predominio de esta bacteria; 2- detectar la producción de fluorescencia en estos medios de cultivo como método rápido de identificación. Los medios de cultivo estudiados fueron: medio selectivo para Veillonella, agar Schaedler para anaerobios con vitamina K, agar tioglicolato, agar infusión cerebro corazón, agar Brucella, agar tripteína soja y agar Columbia con y sin el agregado de vancomicina y sangre lacada. La muestra ensayada fue un pool de saliva. Se hicieron recuentos de colonias de veillonellas y de microorganismos totales expresados en UFC/ml de saliva. La mayor recuperación de veillonellas en saliva se obtuvo en el medio selectivo para Veillonella con vancomicina y sangre lacada. Sólo se observó producción de fluorescencia en este medio.Veillonella spp. are anaerobic gram-negative cocci associated to oral health. Different types of cultures have been reported for the isolation of these microorganisms. Veillonella spp. colonies produce a red fluorescence, which is made visible through ultraviolet light and disappears in contact with oxygen. This feature would be very useful for rapid presumptive identification. The aims of this study were: 1. to compare the Rogosa selective medium for Veillonella with the cultures recommended by different authors in

  6. Kinetics of circulating endogenous insulin, C-peptide, and proinsulin in fasting nondiabetic man

    DEFF Research Database (Denmark)

    Henriksen, Jens Henrik Sahl; Tronier, B; Bülow, J B

    1987-01-01

    Plasma concentrations of insulin, C-peptide, and proinsulin were measured in different vascular beds in order to determine renal, hepatic, and systemic kinetics of the endogenous peptides in the fasting condition. Nineteen nondiabetic subjects were studied, two were normal, nine had minor vascular...

  7. Nicotine concentrations in urine and saliva of smokers and non-smokers.

    Science.gov (United States)

    Feyerabend, C; Higenbottam, T; Russell, M A

    1982-01-01

    Nicotine concentrations were measured in saliva and urine samples collected from 82 smokers and 56 non-smokers after a morning at work. Each subject answered a series of questions related to their recent intentional or passive exposure to tobacco smoke. All non-smokers had measurable amounts of nicotine in both saliva and urine. Those non-smokers who reported recent exposure to tobacco smoke had significantly higher nicotine concentrations (p less than 0.001) than those who had not been exposed; their concentrations overlapped those of smokers who had smoked up to three cigarettes before sampling had the greatest influence on nicotine concentrations (r=0.62 for saliva and r=0.51 for urine). Neither the nicotine for yield of cigarettes nor the self-reported degree of inhalation had any significant effect on nicotine concentrations. PMID:6802384

  8. A novel one-step Helicobacter pylori saliva antigen test.

    Science.gov (United States)

    Yang, Bi-Ling; Yeh, Chun; Kwong, Wei-Gang; Lee, Shou-Dong

    2015-02-01

    A rapid, reliable, and sufficiently accurate test for diagnosing Helicobacter pylori infection is required for screening dyspeptic patients before a referral for endoscopy. The purpose of this article is two-fold: first, to evaluate the accuracy of a one-step H. pylori saliva antigen (HPS) test; and second, to compare noninvasive and invasive H. pylori tests in Taiwanese population. A total of 104 consecutive dyspeptic patients admitted for gastroenterology into the outpatient department underwent a one-step HPS test, rapid urease test, histology, and (13)C-urea breath test (13)C-UBT (proto C-13 urea kit). The accuracy of the HPS test was compared with a gold standard defined by at least two positive H. pylori test results from three H. pylori tests (histology, rapid urease test, and (13)C-UBT). The 104 patients eligible for analysis (mean age: 58 years, range 22-87 years), 21 (20%) were gold standard positive. Among them, the positive of the one-step H. pylori saliva Ag test, rapid urease test, (13)C-UBT, histology were (52; 50%), (17; 16%), (27; 25%) and (22; 21%) respectively. The sensitivity and specificity of the HPS tests, rapid urease test, (13)C-UBTs, and histology were 71.43% and 55.42%, 76.19% and 98.80%, 100% and 92.77%, and 85.71% and 95.18%, respectively, relative to the gold standard. The one-step HPS test exhibited a sensitivity of 71.43%, nearly equivalent to that of the rapid urea test. The one-step HPS test exhibited a high sensitivity and low specificity compared with the other tests, indicating that it is not sufficiently accurate for use in a clinical setting for diagnosing H. pylori infection. However, the test is simple to use (requiring only a saliva sample), inexpensive, and noninvasive in its application, and thus appealing for use in population-based prevalence surveys of the epidemiology of H. pylori infection. Copyright © 2014. Published by Elsevier Taiwan.

  9. Quantitative study of non-stimulated human whole saliva using NAA

    International Nuclear Information System (INIS)

    Lewgoy, H.R.; Zamboni, C.B.; Sabrina Metairon; Medeiros, I.M.M.A.; Medeiros de, J.A.G.

    2013-01-01

    In the past few years, the use of saliva has increased as a method for diagnosis of a variety of diseases. Investigations of flow rates, pH, molecular components, hormones and proteins have presented significant progress in clinical testing as a diagnostic tool. Inorganic elements found in saliva also have important correlations that can assist in the diagnosis of periodontal disease, but these salivary components are still poorly investigated. In this study, we investigated non-stimulated whole saliva of 44 healthy subjects and 12 patients with periodontal disease, obtained from donors at Sao Paulo city (Brazil). Using neutron activation analysis (NAA) technique, we found considerable metabolic changes in the salivary composition of periodontal patients: abnormal concentrations of Br, Ca, I, K, Mg and S that may be associated with periodontal, with the most effective indicator of periodontal disease being Ca concentration. The data from healthy donors also provide a scientific basis for biomedical researches of other oral diseases. (author)

  10. Observations on saliva osmolality during progressive dehydration and partial rehydration.

    Science.gov (United States)

    Taylor, Nigel A S; van den Heuvel, Anne M J; Kerry, Pete; McGhee, Sheena; Peoples, Gregory E; Brown, Marc A; Patterson, Mark J

    2012-09-01

    A need exists to identify dehydrated individuals under stressful settings beyond the laboratory. A predictive index based on changes in saliva osmolality has been proposed, and its efficacy and sensitivity was appraised across mass (water) losses from 1 to 7%. Twelve euhydrated males [serum osmolality: 286.1 mOsm kg(-1) H(2)O (SD 4.3)] completed three exercise- and heat-induced dehydration trials (35.6°C, 56% relative humidity): 7% dehydration (6.15 h), 3% dehydration (with 60% fluid replacement: 2.37 h), repeat 7% dehydration (5.27 h). Expectorated saliva osmolality, measured at baseline and at each 1% mass change, was used to predict instantaneous hydration state relative to mass losses of 3 and 6%. Saliva osmolality increased linearly with dehydration, although its basal osmolality and its rate of change varied among and within subjects across trials. Receiver operating characteristic curves indicated a good predictive power for saliva osmolality when used with two, single-threshold cutoffs to differentiate between hydrated and dehydrated individuals (area under curve: 3% cutoff = 0.868, 6% cutoff = 0.831). However, when analysed using a double-threshold detection technique (3 and 6%), as might be used in a field-based monitor, <50% of the osmolality data could correctly identify individuals who exceeded 3% dehydration. Indeed, within the 3-6% dehydration range, its sensitivity was 64%, while beyond 6% dehydration, this fell to 42%. Therefore, while expectorated saliva osmolality tracked mass losses within individuals, its large intra- and inter-individual variability limited its predictive power and sensitivity, rendering its utility questionable within a universal dehydration monitor.

  11. LACK OF ASSOCIATION BETWEEN HERPESVIRUS DETECTION IN SALIVA AND GINGIVITIS IN HIV‑INFECTED CHILDREN.

    Science.gov (United States)

    Otero, Renata A; Nascimento, Flávia N N; Souza, Ivete P R; Silva, Raquel C; Lima, Rodrigo S; Robaina, Tatiana F; Câmara, Fernando P; Santos, Norma; Castro, Gloria F

    2015-01-01

    The aims of this study were to compare the detection of human herpesviruses (HHVs) in the saliva of HIV-infected and healthy control children, and to evaluate associations between viral infection and gingivitis and immunodeficiency. Saliva samples were collected from 48 HIV-infected and 48 healthy control children. Clinical and laboratory data were collected during dental visits and from medical records. A trained dentist determined gingival indices and extension of gingivitis. Saliva samples were tested for herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), varicella zoster virus (VZV), Epstein-Barr virus (EBV), and cytomegalovirus (CMV) by nested polymerase chain reaction assays. Thirty-five HIV-infected and 16 control children had gingivitis. Seventeen (35.4%) HIV-infected children and 13 (27%) control children were positive for HHVs. CMV was the most commonly detected HHV in both groups (HIV-infected, 25%; control, 12.5%), followed by HSV-1 (6.2% in both groups) and HSV-2 (HIV-infected, 4.2%; control, 8.3%). The presence of HHVs in saliva was not associated with the presence of gingivitis in HIV-1-infected children (p = 0.104) or healthy control children (p = 0.251), or with immunosuppression in HIV-infected individuals (p = 0.447). Gingivitis was correlated with HIV infection (p = 0.0001). These results suggest that asymptomatic salivary detection of HHVs is common in HIV-infected and healthy children, and that it is not associated with gingivitis.

  12. Características de la saliva en niños con Síndrome de Down

    OpenAIRE

    Alicia Leonor Pinzón-Te; Rubén López-Pérez; Félix Manuel Enríquez-Rivera; Fernando Javier Aguilar- Ayala; María Eduviges Rejón-Peraza; José Ladislao López-Osorio

    2017-01-01

    ANTECEDENTES: los niños con Síndrome de Down (SD) generalmente tienen características morfogenéticas que los predisponen a patologías bucales; por lo tanto, el análisis de las características de la saliva constituye una herramienta diagnóstica de su estado de salud bucal. OBJETIVOS: a) Comparar los valores promedio de pH salival y tasa de flujo salival (TFS), según género y edad. b) Comparar los tipos de pH salival y TFS, según género y edad. c) Determinar si existe correlación entre el ...

  13. Skim Milk, Whey, and Casein Increase Body Weight and Whey and Casein Increase the Plasma C-Peptide Concentration in Overweight Adolescents12

    DEFF Research Database (Denmark)

    Arnberg, Karina; Mølgaard, Christian; Michaelsen, Kim Fleischer

    2012-01-01

    insulin, and insulin secretion estimated as the plasma C-peptide concentration in overweight adolescents. Overweight adolescents (n = 203) aged 12–15 y with a BMI of 25.4 ± 2.3 kg/m2 (mean ± SD) were randomized to 1 L/d of skim milk, whey, casein, or water for 12 wk. All milk drinks contained 35 g protein....... Outcomes were BMI-for-age Z-scores (BAZs), waist circumference, plasma insulin, homeostatic model assessment, and plasma C-peptide. We found no change in BAZ in the pretest control and water groups, whereas it was greater at 12 wk in the skim milk, whey, and casein groups compared with baseline...... and with the water and pretest control groups. The plasma C-peptide concentration increased from baseline to wk 12 in the whey and casein groups and increments were greater than in the pretest control (P

  14. Comparison of Plasma, Saliva, and Hair Levetiracetam Concentrations.

    Science.gov (United States)

    Karaś-Ruszczyk, Katarzyna; Kuczyńska, Julita; Sienkiewicz-Jarosz, Halina; Kurkowska-Jastrzębska, Iwona; Bienkowski, Przemyslaw; Restel, Magdalena; Samochowiec, Jerzy; Mierzejewski, Pawel

    2017-06-01

    Previous findings revealed high correlations between serum/plasma and saliva levetiracetam concentrations, indicating saliva as an alternative matrix for monitoring levetiracetam therapy. Levetiracetam concentration in the hair, which could reflect long-term drug exposure and patients' compliance, has not been systematically tested, as yet. The aim of this study was to determine the correlation between plasma, saliva, and hair levetiracetam concentrations in 47 patients with epilepsy. Plasma, saliva, and hair levetiracetam concentrations were measured by liquid chromatography-tandem mass spectrometry with positive ionization. Levetiracetam saliva and plasma concentrations were highly correlated (r = 0.93). Plasma concentrations were not influenced by sex, age, and other concomitant antiepileptic drugs. Levetiracetam hair concentrations correlated with plasma concentrations (r = 0.36) but not daily dose (mg/kg). Drug hair concentrations were not influenced by hair color or treatment (dyed). The results tend to indicate that saliva may be a reliable alternative to plasma for monitoring levetiracetam concentrations. Levetiracetam can also be detected in human hair.

  15. Comparison of two extraction procedures for determination of drugs of abuse in human saliva by high-performance liquid chromatography.

    Science.gov (United States)

    Fernández, P; Morales, L; Vázquez, C; Lago, M; Bermejo, A M

    2008-11-01

    High performance liquid chromatography in combination with diode array detection (HPLC-DAD) was used to determine morphine, 6-acetylmorphine, cocaine, benzoylecgonine, cocaethylene, methadone and 2-ethylene-1,5-dimethyl-3,3,-diphenylpyrrolidine in human saliva. For comparison, samples were prepared by either liquid-liquid extraction in Toxitubes A or microwave-assisted extraction (MAE), by mixing 1 ml of saliva with 10 ml of chloroform and operating at 100 degrees C for 10 min. Acetonitrile and 0.02 m phosphate buffer at pH 6.5 were used as mobile phase in HPLC in gradient mode. The detector response was linear over the drug concentration range of 0.05-2.0 microg ml(-1) in human saliva. The analytical method was validated by determining its precision and accuracy (n = 5), which were lower than 5% as relative standard deviation and 6% as relative error. Limits of detection ranged from 10 to 35 ng ml(-1); mean recoveries of drugs were from 53 to 95% with Toxitubes A and from 83 to 100% with MAE at two different concentrations (0.1 and 1.0 microg ml(-1)). The proposed method was applied to 24 saliva samples from individuals poisoned with opiates and/or cocaine.

  16. Purification and characterization of insulin and the C-peptide of proinsulin from Przewalski's horse, zebra, rhino, and tapir (Perissodactyla).

    Science.gov (United States)

    Henry, J S; Lance, V A; Conlon, J M

    1993-02-01

    Within the order Perissodactyla, the primary structure of insulin has been strongly conserved. Insulin from Przewalski's horse and the mountain zebra (suborder Hippomorpha) is the same as that from the domestic horse and differs from insulin from the white rhinoceros and mountain tapir (suborder Ceratomorpha) by a single substitution (Gly-->Ser) at position 9 in the A-chain. A second molecular form of Przewalski's horse insulin isolated in this study was shown to represent the gamma-ethyl ester of the Glu17 residue of the A-chain. This component was probably formed during the extraction of the pancreas with acidified ethanol. The amino acid sequence of the C-peptide of proinsulin has been less well conserved. Zebra C-peptide comprises 31 amino acid residues and differs from Przewalski's horse and domestic horse C-peptide by one substitution (Gln30-->Pro). Rhino C-peptide was isolated only in a truncated form corresponding to residues (1-23) of intact C-peptide. Its amino acid sequence contains three substitutions compared with the corresponding region of horse C-peptide. It is postulated that the substitution (Pro23-->Thr) renders rhino C-peptide more liable to proteolytic cleavage by a chymotrypsin-like enzyme than horse C-peptide. C-peptide could not be identified in the extract of tapir pancreas, suggesting that proteolytic degradation may have been more extensive than in the rhino. In contrast to the ox and pig (order Artiodactyla), there was no evidence for the expression of more than one proinsulin gene in the species of Perissodactyla examined.

  17. Inhibition of the classical pathway of the complement system by saliva of Amblyomma cajennense (Acari: Ixodidae).

    Science.gov (United States)

    Franco, Paula F; Silva, Naylene C S; Fazito do Vale, Vladimir; Abreu, Jéssica F; Santos, Vânia C; Gontijo, Nelder F; Valenzuela, Jesus G; Pereira, Marcos H; Sant'Anna, Mauricio R V; Gomes, Alessandra P S; Araujo, Ricardo N

    2016-05-01

    Inhibition of the complement system during and after haematophagy is of utmost importance for tick success in feeding and tick development. The role of such inhibition is to minimise damage to the intestinal epithelium as well as avoiding inflammation and opsonisation of salivary molecules at the bite site. Despite its importance, the salivary anti-complement activity has been characterised only in species belonging to the Ixodes ricinus complex which saliva is able to inhibit the alternative and lectin pathways. Little is known about this activity in other species of the Ixodidae family. Thus, the aim of this study was to describe the inhibition of the classical pathway of the complement system by the saliva of Amblyomma cajennense at different stages of the haematophagy. The A. cajennense saliva and salivary gland extract (SGE) were able to inhibit the complement classical pathway through haemolytic assays with higher activity observed when saliva was used. The anti-complement activity is present in the salivary glands of starving females and also in females throughout the whole feeding process, with significant higher activity soon after tick detachment. The SGE activity from both females fed on mice or horses had no significant correlation (p > 0.05) with tick body weight. The pH found in the intestinal lumen of A. cajennense was 8.04 ± 0.08 and haemolytic assays performed at pH 8.0 showed activation of the classical pathway similarly to what occurs at pH 7.4. Consequently, inhibition could be necessary to protect the tick enterocytes. Indeed, the inhibition observed by SGE was higher in pH 8.0 in comparison to pH 7.4 reinforcing the role of saliva in protecting the intestinal cells. Further studies should be carried out in order to identify the inhibitor molecule and characterise its inhibition mechanism. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. Natriuretic peptides stimulate the cardiac sodium pump via NPR-C-coupled NOS activation

    DEFF Research Database (Denmark)

    William, M.; Hamilton, E.J.; Garcia, A.

    2008-01-01

    Natriuretic peptides (NPs) and their receptors (NPRs) are expressed in the heart, but their effects on myocyte function are poorly understood. Because NPRs are coupled to synthesis of cGMP, an activator of the sarcolemmal Na(+)-K(+) pump, we examined whether atrial natriuretic peptide (ANP) regul...

  19. An improved method of 18F peptide labeling: hydrazone formation with HYNIC-conjugated c(RGDyK)

    International Nuclear Information System (INIS)

    Lee, Yun-Sang; Jeong, Jae Min; Kim, Hyung Woo; Chang, Young Soo; Kim, Young Joo; Hong, Mee Kyung; Rai, Ganesha B.; Chi, Dae Yoon; Kang, Won Jun; Kang, Joo Hyun; Lee, Dong Soo; Chung, June-Key; Lee, Myung Chul; Suh, Young-Ger

    2006-01-01

    Radiolabeled α v β 3 -integrin antagonists are increasingly investigated as a means of imaging angiogenesis. Several methods of labeling α v β 3 -integrin binding peptide with 18 F have been reported recently. In the present study, we devised a straightforward means for labeling Arg-Gly-Asp (RGD) peptide with 18 F via hydrazone formation between c(RGDyK)-hydrazinonicotinic acid (HYNIC) (3) and 4-[ 18 F]-fluorobenzaldehyde ([ 18 F]4). The resulting reaction mixture was purified by HPLC to give 4'-[ 18 F]-fluorobenzylidenehydrazone-6-nicotinamide-c(RGDyK) ([ 18 F]5). The conjugation efficiency of 3 and 4 to form [ 18 F]5 was 95.2%, and the radiochemical purity of [ 18 F]5 after purification was >99%. The specific activity of [ 18 F]5 estimated by radio-HPLC was 20.5 GBq/μmol (end of synthesis). Competitive binding assay of c(RGDyK) (1) and 5 was performed using [ 125 I]iodo-c(RGDyK) as a radioligand, and K i values were found to be 2.8 and 21.7 nM, respectively. For the biodistribution study, the angiogenic mouse model was established by inducing unilateral ischemia on the left hindlimbs of ICR mice after femoral artery ablation. Seven days after inducing ischemia, [ 18 F]5 was administered to the mice through the tail vein. Ischemic muscle uptake of [ 18 F]5 was significantly higher than that of normal muscle (P 18 F]5. Here, we successfully labeled RGD peptide with 18 F via hydrazone formation between 3 and 4, resulting to [ 18 F]5. [ 18 F]5 was found to have high affinity for α v β 3 -integrin and to accumulate specifically in ischemic hindlimb muscle of mice. We suggest that 18 F labeling via formation of hydrazone between HYNIC peptide and [ 18 F]4 is a useful method for labeling c(RGDyK), which can be applied for imaging angiogenesis

  20. Is parotid saliva sterile on entry to the oral cavity?

    DEFF Research Database (Denmark)

    Schrøder, Stine A; Bardow, Allan; Eickhardt-Dalbøge, Steffen

    2017-01-01

    CONCLUSION: The present study indicates that parotid saliva is sterile on entry to the oral cavity. OBJECTIVES: The objective was to investigate if parotid saliva is sterile on entry to the oral cavity and, thus, prior to contamination by oral bacteria. METHOD: Forty healthy volunteers were...... included in sterile parotid saliva collection. Parotid saliva was collected using a sterile Lashley cup, placed over the papilla of the Stensen´s duct, as well as sterile tubes and syringes for collection. All collections were followed by collection of a positive control sample where some of the sterile...... obtained parotid saliva had been exposed to the contralateral mucosal membranes. All samples parotid saliva, as well as the positive controls, were cultivated, and 10 randomly selected parotid saliva samples underwent polymerase chain reaction (PCR) analyses. RESULTS: In 33 of 40 parotid saliva samples...

  1. Reversible pH-controlled DNA-binding peptide nanotweezers: An in-silico study

    Directory of Open Access Journals (Sweden)

    Gaurav Sharma

    2008-11-01

    Full Text Available Gaurav Sharma1, Kaushal Rege2,3, David E Budil4, Martin L Yarmush2,5, Constantinos Mavroidis11Department of Mechanical and Industrial Engineering; 4Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA, USA; 2The Center for Engineering in Medicine (CEM, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA; 3Department of Chemical Engineering, Arizona State University, Tempe, AZ, USA; 5Department of Biomedical Engineering, Rutgers University, NJ, USAAbstract: We describe the molecular dynamics (MD-aided engineering design of mutant peptides based on the α-helical coiled-coil GCN4 leucine zipper peptide (GCN4-p1 in order to obtain environmentally-responsive nanotweezers. The actuation mechanism of the nanotweezers depends on the modification of electrostatic charges on the residues along the length of the coiled coil. Modulating the solution pH between neutral and acidic values results in the reversible movement of helices toward and away from each other and creates a complete closed-open-closed transition cycle between the helices. Our results indicate that the mutants show a reversible opening of up to 15 Å (1.5 nm; approximately 150% of the initial separation upon pH actuation. Investigation on the physicochemical phenomena that influence conformational properties, structural stability, and reversibility of the coiled-coil peptide-based nanotweezers revealed that a rationale- and design-based approach is needed to engineer stable peptide or macromolecules into stimuli-responsive devices. The efficacy of the mutant that demonstrated the most significant reversible actuation for environmentally responsive modulation of DNA-binding activity was also demonstrated. Our results have significant implications in bioseparations and in the engineering of novel transcription factors.Keywords: bionanotechnology, nanotweezers, coiled-coil, GCN4, leucine zipper, molecular dynamics, environmentally

  2. Preparation and characterization of different liposomal formulations containing P5 HER2/neu-derived peptide and evaluation of their immunological responses and antitumor effects

    Directory of Open Access Journals (Sweden)

    Sheida Shariat

    2015-05-01

    Full Text Available Objective(s:Tumor-associated antigen (TAA subunit-based vaccines constitute promising tools for anticancer immunotherapy. However, a major limitation in the development of such vaccines is the poor immunogenicity of peptides when used alone.The aim of this study was to develop an efficient vaccine delivery system and adjuvant to enhance anti-tumor activity of a synthetic HER2/neu derived peptide (P5. Materials and Methods: P5 peptide was encapsulated with different liposomal formulations composed of DMPC:DMPG:Chol:DOPE and loaded with monophosphoryl lipid A (MPL. All formulations were characterized for their physicochemical properties. To evaluate vaccine efficacy, BALB/c mice were first immunized with free peptide or liposomal formulations, then, inoculated with a subcutaneous injection of TUBO tumor cells. Enzyme-linked immunospot, cytotoxicity and intracellular cytokine assays, as well as tumor size and animal survival analysis, were performed to evaluate the immune responses. Results: The results demonstrated that P5 encapsulated into liposomal formulations was not able to induce CD8 and CD4 T cells to produce IFN-γ. That is why, a potent CTL response and antitumor immunity was not induced. Conclusion: The Lip-DOPE-P5-MPL formulation in spite of using pH-sensitive lipid to direct intracellular trafficking of peptide to MHC I presentation pathway and MPL to enhance peptide adjuvanticity was interesting. The failure in inducing anti-tumor immunity may be attributed to low uptake of anionic conventional liposomes by dendritic cells (DCs that have negative surface charge.

  3. Quantification of piroxicam and 5'-hydroxypiroxicam in human plasma and saliva using liquid chromatography-tandem mass spectrometry following oral administration.

    Science.gov (United States)

    Calvo, Adriana Maria; Santos, Gabriel Mulinari; Dionísio, Thiago José; Marques, Maria Paula; Brozoski, Daniel Thomas; Lanchote, Vera Lúcia; Fernandes, Maria Helena Raposo; Faria, Flávio Augusto Cardoso; Santos, Carlos Ferreira

    2016-02-20

    Saliva sampling used to quantify piroxicam and 5'-hydroxypiroxicam is a noninvasive and painless method when compared to sequential blood sampling. For that, a rapid, selective and sensitive liquid chromatography-tandem mass spectrometric method for simultaneous determination of piroxicam and 5'-hydroxypiroxicam in saliva and human plasma was developed and validated. Piroxicam and its major metabolite were separated using a LiChroCART 125-4 RP Select-B Sorbent C18 column using a mixture of methanol and 2% phosphoric acid (pH 2.7) (70:30, v/v) for the mobile phase with a flow injection of 1mL/min. The run time was 4min. Volunteers had saliva and blood sampled before, 1, 2, 3, 4, 5, 6, 8, 11, 24, 48 and 72h after taking a 20mg oral dose of piroxicam. The pharmacokinetic parameters of piroxicam in plasma samples were as follows: AUC0-72 (64819hng/mL), predicted clearance (0.2L/h), distribution volume (14.8L), elimination half-life (50.7h) and saliva/plasma concentration ratio (0.003). The estimation of all pharmacokinetic parameters for 5'-hydroxypiroxicam would require collections beyond 72h; however, it was possible to quantify the mean maximum concentration (133ng/mL), time to peak concentration (53.6h), mean AUC0-72 (6213hng/mL), predicted clearance (110.3L/h) and saliva/plasma concentration ratio (0.04). The developed methods proved effective and sensitive for determining the lower quantification limit of piroxicam in plasma (6.1ng/mL) and saliva (0.15ng/mL) and of 5'-hydroxypiroxicam in plasma (1.2ng/mL) and saliva (0.15ng/mL). Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Human Saliva Collection Devices for Proteomics: An Update

    Directory of Open Access Journals (Sweden)

    Zohaib Khurshid

    2016-06-01

    Full Text Available There has been a rapid growth in the interest and adaptation of saliva as a diagnostic specimen over the last decade, and in the last few years in particular, there have been major developments involving the application of saliva as a clinically relevant specimen. Saliva provides a “window” into the oral and systemic health of an individual, and like other bodily fluids, saliva can be analyzed and studied to diagnose diseases. With the advent of new, more sensitive technologies to detect smaller concentrations of analytes in saliva relative to blood levels, there have been a number of critical developments in the field that we will describe. In particular, recent advances in standardized saliva collection devices that were not available three to four years ago, have made it easy for safe, simple, and non-invasive collection of samples to be carried out from patients. With the availability of these new technologies, we believe that in the next decade salivary proteomics will make it possible to predict and diagnose oral as well as systemic diseases, cancer, and infectious diseases, among others. The aim of this article is to review recent developments and advances in the area of saliva specimen collection devices and applications that will advance the field of proteomics.

  5. Salivary flow rate and biochemical composition analysis in stimulated whole saliva of children with cystic fibrosis.

    Science.gov (United States)

    da Silva Modesto, Karine Barros; de Godói Simões, Jéssica Bueno; de Souza, Amanda Ferreira; Damaceno, Neiva; Duarte, Danilo Antonio; Leite, Mariana Ferreira; de Almeida, Eliete Rodrigues

    2015-11-01

    It is recognized that cystic fibrosis (CF) patients present a risk for oral diseases, since it affects exocrine glands, and the treatment consists of a carbohydrate-rich diet. Recognizing the protective function of saliva on maintaining oral health, the aim of the study was to evaluate salivary parameters in stimulated whole saliva from children with CF. A case-control study was conducted comparing stimulated whole saliva of healthy (n=28; control group) and CF children (n=21; experimental group). Salivary flow rate, initial pH, buffer capacity (total and in each range of pH), total protein and sialic acid (total, free, and conjugated) concentration, α-amylase and salivary peroxidase activities were evaluated. Data were compared by two-tailed Student t test (95% CI; p ≤ 0.05). CF patients presented a significant reduction in salivary parameters compared with the control group (p ≤ 0.05): salivary flow rate (36%), buffer capacity (pH range from 6.9 to 6.0), sialic acid concentration (total 75%, free 61%, and conjugated 83%); α-amylase and salivary peroxidase activities (55%). Additionally, a significant increase in total protein concentration (180%) of stimulated whole saliva from CF patients was verified compared with the control group (p ≤ 0.05). Children with CF presented significant changes in salivary composition, including salivary flow rate, buffering capacity and protective proteins of the oral cavity, compared with children without CF. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Identification and accurate quantification of structurally related peptide impurities in synthetic human C-peptide by liquid chromatography-high resolution mass spectrometry.

    Science.gov (United States)

    Li, Ming; Josephs, Ralf D; Daireaux, Adeline; Choteau, Tiphaine; Westwood, Steven; Wielgosz, Robert I; Li, Hongmei

    2018-06-04

    Peptides are an increasingly important group of biomarkers and pharmaceuticals. The accurate purity characterization of peptide calibrators is critical for the development of reference measurement systems for laboratory medicine and quality control of pharmaceuticals. The peptides used for these purposes are increasingly produced through peptide synthesis. Various approaches (for example mass balance, amino acid analysis, qNMR, and nitrogen determination) can be applied to accurately value assign the purity of peptide calibrators. However, all purity assessment approaches require a correction for structurally related peptide impurities in order to avoid biases. Liquid chromatography coupled to high resolution mass spectrometry (LC-hrMS) has become the key technique for the identification and accurate quantification of structurally related peptide impurities in intact peptide calibrator materials. In this study, LC-hrMS-based methods were developed and validated in-house for the identification and quantification of structurally related peptide impurities in a synthetic human C-peptide (hCP) material, which served as a study material for an international comparison looking at the competencies of laboratories to perform peptide purity mass fraction assignments. More than 65 impurities were identified, confirmed, and accurately quantified by using LC-hrMS. The total mass fraction of all structurally related peptide impurities in the hCP study material was estimated to be 83.3 mg/g with an associated expanded uncertainty of 3.0 mg/g (k = 2). The calibration hierarchy concept used for the quantification of individual impurities is described in detail. Graphical abstract ᅟ.

  7. Three-dimensional structure and dynamics of wine tannin-saliva protein complexes. A multitechnique approach.

    Science.gov (United States)

    Simon, Cécile; Barathieu, Karine; Laguerre, Michel; Schmitter, Jean-Marie; Fouquet, Eric; Pianet, Isabelle; Dufourc, Erick J

    2003-09-09

    The interactions between the B3 (catechin-4alpha,8-catechin) red wine tannin and the human salivary protein fragment IB7(14) (SPPGKPQGPPPQGG) were monitored by (1)H magic angle spinning NMR, circular dichroism, electrospray ionization mass spectrometry, and molecular modeling. It is found that the secondary structure of IB7(14) is made of a type II helix (collagen helix) and random coil. The central glycine 8 appears to act as a flexible rotula separating two helix II regions. Three tannin molecules tightly complex the peptide, without modifying its secondary structure, but seem to reduce its conformational dynamics. The binding dissociation constant is in the millimolar range. B3 tannins with a "tweezers" conformation bind to the hydrophilic side of the saliva peptide, suggesting that the principal driving forces toward association are governed by hydrogen bonding between the carbonyl functions of proline residues and both the phenol and catechol OH groups. These findings are further discussed in the frame of an astringency phenomenon.

  8. Saliva as a diagnostic fluid: literature review

    OpenAIRE

    Martí Álamo, Silvia; Mancheño Franch, Aisha; Marzal Gamarra, Cristina

    2012-01-01

    There is a growing interest in diagnosis based on the analysis of saliva. This is a simple, non-invasive method of obtaining oral samples which is safe for both the health worker and the patient, not to mention allowing for simple and cost-efficient storage. The majority of studies use general saliva samples in their entirety, complex fluids containing both local and systemic sources and whose composition corresponds to that of the blood. General saliva contains a considerable ...

  9. State of the Art: Saliva-Hormonanalyse // State of the art: Saliva analysis

    Directory of Open Access Journals (Sweden)

    Obermayer-Pietsch B

    2017-01-01

    Full Text Available Saliva analyses are widely used for hormone measurements, but also for drug and therapy monitoring, periodontal inflammation and human, viral or microbial nucleic acid (DNA detection. In this review, we focus on preanalytics, and measurement techniques. Clinical applications of steroid hormone analyses are presented with regard to practical use. p bKurzfassung: /bSpeichelanalysen können Hormone, aber auch Drogen- und Medikamenten-Monitoring, parodontale Entzündungsparameter und humane, virale oder mikrobielle Nukleinsäuren (DNA umfassen. In dieser Übersichtsarbeit wird auf Präanalytik, Nachweis-Techniken und Beispiele u.a. aus der Steroidanalyse eingegangen, die derzeit am weitesten entwickelt ist und vielerorts bereits Eingang in die medizinische Routine findet.

  10. Effect of Immersion Time in Artificial Saliva on Flexural Strength of Provisional Crown and Bridge Material: Light zPolymerization versus Autopolymerization system

    Directory of Open Access Journals (Sweden)

    Marzia Magdalena Tetelepta

    2013-07-01

    Full Text Available Objective: The aim of this study was to investigate the effect of immersion time in artificial salive on the flexural strength of provisional crown and bridge (p-c&b materials. Materials and Methods: Two types of p-c&b materials were used in this study: Light polymerized p-c&b material (Revotek LC and autopolymerized p-c&b material (PerfecTemp II. A total of 100 specimens were fabricated and measured according to ISO 4049/2000. A stainless steel mould was used to prepare 2mmx2mmx25mm bar shaped specimens. All materials were dispensed and manipulated according to the manufacturers' instructions. The specimens were divided into 5 groups (n=10. Each specimen of the first group was measured immediately after preparation. The second, third, fourth and fifth groups were immersed in artificial saliva at 37ºC in an incubator for 1 hour, 1 day, 7 days, and 14 days, respectively. Flexural strength was tested by Universal Mechanical Testing Machine Shimadzu in a 3-point bending test. The repeated ANOVA and Post-Hoc Bonferroni test were used to compare the continuous variables between the groups. Results: The results showed flexural strength of Revotek LC were higher than PerfecTemp II at first and second group. However, flexural strength of PerfecTemp II was higher than Revotek LC at third, fourth, and fifth group. The highest flexural strength of Revotek LC was achieved in 1 hour immersion, whereas PerfecTemp II achieved the highest value in 7 days. Conclusion: Flexural strength of p-c&b materials were influenced by immersion time in artifical saliva and the type of p-c&b materials.DOI: 10.14693/jdi.v17i1.108

  11. Corrosion of Nickel-Titanium Orthodontic Archwires in Saliva and Oral Probiotic Supplements

    Directory of Open Access Journals (Sweden)

    Gianluca Turco

    2017-01-01

    Full Text Available Objectives: The aim of the study was to examine how probiotic supplements affect the corrosion stability of orthodontic archwires made of nickel-titanium alloy (NiTi. Materials and Methods: Ni-Ti archwires (0.508x0.508 and having the length of 2.5 cm were tested. The archwires (composition Ni=50.4%, Ti=49.6% were uncoated, nitrified and rhodium coated. Surface microgeometry was observed by using scanning electron microscope and surface roughness was measured by profilometer through these variables: roughness average, maximum height and maximum roughness depth. Corrosion was examined by electrochemical method of cyclic polarisation. Results: Rhodium coated alloy in saliva has significantly higher general corrosion in saliva than nitrified alloy and uncoated alloy, with large effect size (p=0.027; η2=0.700. In the presence of probiotics, the result was even more pronounced (p<0.001; η2=0.936. Probiotic supplement increases general and localised corrosion of rhodium coated archwire and slightly decreases general corrosion and increases localised corrosion in uncoated archwire , while in the case of nitrified archwire the probability of corrosion is very low. The differences in surface roughness between NiTi wires before corrosion are not significant. Exposure to saliva decreases roughness average in rhodium coated wire (p=0.015; η2=0.501. Media do not significantly influence surface microgeometry in nitrified and uncoated wires. Conclusion: Probiotic supplement affects corrosion depending on the type of coating of the NiTi archwire. It increases general corrosion of rhodium coated wire and causes localised corrosion of uncoated and rhodium coated archwire. Probiotic supplement does not have greater influence on surface roughness compared to that of saliva.

  12. Radioimmunological analysis of circadian rhythms of cortisol and melatonin in saliva

    International Nuclear Information System (INIS)

    Demel, A.W.

    1990-12-01

    Since blood cortisol (F) and melatonin (MLT) display a circadian secretion pattern and since the saliva concentration of this hormones is an excellent indicator of its blood levels the measurement of salivary F and MLT may be user for examining circadian rhythmicity. In this study the relationship between salivary F and MLT was explored. For this purpose it was necessary first to establish and validate a radioimmunoassay for F in saliva: salivary F was determined by a direct radioimmunoassay using cortisol-3-(O-carb oxymethyl) oximino-(2-( 125 I)iodohistamin) as tracer and cortisol-3-CMO-BSA antiserum. The parallel measurement of F levels in saliva and serum of adults gave an excellent correlation (r=0.87, p 0.00956x ). Serum F was assayed on the Abott TDX-System using a radioimmunofluorescence method. Secondly, using this assay the circadian saliva F pattern was determined as well as the pattern of salivary MLT in 9 young, healthy volunteers. For saliva MLT estimations a previously published method was applied (SCHULZ et al 1990). Using a computerized program (RHYTHM) written by EVE v. CAUTER (1979), the hormone data of each individuum were examined for circadian rhythmicity and its acrophase (time of occurence of the maximum of a sinusoid fitted to the data). The F acrophase occured between 7:00 and 12:00 h (Mean: 3:33 h, SD: 104.4 min). The easy stress-free non invasive nature of saliva collection makes saliva to one of the most accessible body fluids and of high value in studying the circadian system in healthy humans as well as in infants, children, pregnant women and anaemic patients. Measurements of salivary F and MLT may help to elucidate not only the circadian rhythms of these hormones under normal and pathological conditions but it may also provide insight in physiology and pathology of the circadian system in general. (author)

  13. Parámetros inflamatorios en saliva y sangre en niños y adolescentes sanos Inflammatory parameters in saliva and blood from healthy children and adolescents

    Directory of Open Access Journals (Sweden)

    Ninoska Tahis Viera Sirit

    2011-09-01

    Full Text Available En la actualidad se ha mostrado interés en el empleo de la saliva para ser utilizada como una alternativa de diagnóstico, predicción y progresión de diversas enfermedades con relación a otros fluidos corporales. Los objetivos trazados para la realización de este trabajo fueron: correlacionar las concentraciones en saliva y sangre de IL-1, IL-6, TNF-a, sustancias reactivas al ácido tiobarbitúrico y O2- de niños y adolescentes sistémicamente sanos. Se realizó un estudio de corte transversal en 23 niños y adolescentes sanos, entre 4 y 17 años de edad. Se les realizaron evaluaciones clínicas para determinar las condiciones bucales y estudios inmunológicos con el propósito de identificar los niveles de citosinas, a través del ensayo inmunoenzimático indirecto, el O2- por método citoquímico y las sustancias reactivas al ácido tiobarbitúrico, a través del ensayo colorimétrico. Hubo diferencia significativa entre las muestras de saliva y las de sangre periférica respecto a las citosinas y sustancias reactivas al ácido tiobarbitúrico estudiadas. Los resultados fueron: IL-1 en sangre= 1,646 ± 0,13 pg/mL y de IL-1 en saliva= 552,36 ± 75,7 pg/mL; IL-6 en sangre= 3,506 ± 1,85 pg/mL, e IL-6 en saliva= 26,89 ± 9,97 pg/mL. Al analizar el TNF-a en sangre fue de 12,91 ± 3,05 pg/mL y en saliva= 43,56 ± 6,44 pg/mL, las sustancias reactivas al ácido tiobarbitúrico en sangre= 9,46 ± 3,26 nmol/mL y en saliva= 1,26 ± 0,03 nmol/mL. No se observó correlación estadísticamente significativa entre las muestras de sangre y saliva para los valores de IL-1, IL-6 y sustancias reactivas al ácido tiobarbitúrico. En cuanto al TNF-a se evidenció una correlación significativa, r s= 0,78. No se evidenciaron células positivas para el O2- en las muestras estudiadas. Los resultados del análisis de correlación obtenido entre las muestras salivales y séricas, no aportaron evidencias suficientes para sugerir que la saliva pueda ser utilizada

  14. Occurrence of C-Terminal Residue Exclusion in Peptide Fragmentation by ESI and MALDI Tandem Mass Spectrometry

    Science.gov (United States)

    Dupré, Mathieu; Cantel, Sonia; Martinez, Jean; Enjalbal, Christine

    2012-02-01

    By screening a data set of 392 synthetic peptides MS/MS spectra, we found that a known C-terminal rearrangement was unexpectedly frequently occurring from monoprotonated molecular ions in both ESI and MALDI tandem mass spectrometry upon low and high energy collision activated dissociations with QqTOF and TOF/TOF mass analyzer configuration, respectively. Any residue localized at the C-terminal carboxylic acid end, even a basic one, was lost, provided that a basic amino acid such arginine and to a lesser extent histidine and lysine was present in the sequence leading to a fragment ion, usually depicted as (bn-1 + H2O) ion, corresponding to a shortened non-scrambled peptide chain. Far from being an epiphenomenon, such a residue exclusion from the peptide chain C-terminal extremity gave a fragment ion that was the base peak of the MS/MS spectrum in certain cases. Within the frame of the mobile proton model, the ionizing proton being sequestered onto the basic amino acid side chain, it is known that the charge directed fragmentation mechanism involved the C-terminal carboxylic acid function forming an anhydride intermediate structure. The same mechanism was also demonstrated from cationized peptides. To confirm such assessment, we have prepared some of the peptides that displayed such C-terminal residue exclusion as a C-terminal backbone amide. As expected in this peptide amide series, the production of truncated chains was completely suppressed. Besides, multiply charged molecular ions of all peptides recorded in ESI mass spectrometry did not undergo such fragmentation validating that any mobile ionizing proton will prevent such a competitive C-terminal backbone rearrangement. Among all well-known nondirect sequence fragment ions issued from non specific loss of neutral molecules (mainly H2O and NH3) and multiple backbone amide ruptures (b-type internal ions), the described C-terminal residue exclusion is highly identifiable giving raise to a single fragment ion in

  15. P17, an Original Host Defense Peptide from Ant Venom, Promotes Antifungal Activities of Macrophages through the Induction of C-Type Lectin Receptors Dependent on LTB4-Mediated PPARγ Activation

    Directory of Open Access Journals (Sweden)

    Khaddouj Benmoussa

    2017-11-01

    Full Text Available Despite the growing knowledge with regard to the immunomodulatory properties of host defense peptides, their impact on macrophage differentiation and on its associated microbicidal functions is still poorly understood. Here, we demonstrated that the P17, a new cationic antimicrobial peptide from ant venom, induces an alternative phenotype of human monocyte-derived macrophages (h-MDMs. This phenotype is characterized by a C-type lectin receptors (CLRs signature composed of mannose receptor (MR and Dectin-1 expression. Concomitantly, this activation is associated to an inflammatory profile characterized by reactive oxygen species (ROS, interleukin (IL-1β, and TNF-α release. P17-activated h-MDMs exhibit an improved capacity to recognize and to engulf Candida albicans through the overexpression both of MR and Dectin-1. This upregulation requires arachidonic acid (AA mobilization and the activation of peroxisome proliferator-activated receptor gamma (PPARγ nuclear receptor through the leukotriene B4 (LTB4 production. AA/LTB4/PPARγ/Dectin-1-MR signaling pathway is crucial for P17-mediated anti-fungal activity of h-MDMs, as indicated by the fact that the activation of this axis by P17 triggered ROS production and inflammasome-dependent IL-1β release. Moreover, we showed that the increased anti-fungal immune response of h-MDMs by P17 was dependent on intracellular calcium mobilization triggered by the interaction of P17 with pertussis toxin-sensitive G-protein-coupled receptors on h-MDMs. Finally, we also demonstrated that P17-treated mice infected with C. albicans develop less severe gastrointestinal infection related to a higher efficiency of their macrophages to engulf Candida, to produce ROS and IL-1β and to kill the yeasts. Altogether, these results identify P17 as an original activator of the fungicidal response of macrophages that acts upstream PPARγ/CLRs axis and offer new immunomodulatory therapeutic perspectives in the field of

  16. Disposable Collection Kit for Rapid and Reliable Collection of Saliva

    OpenAIRE

    Yamaguchi, Masaki; Tezuka, Yuki; Takeda, Kazunori; Shetty, Vivek

    2015-01-01

    Objectives To describe and evaluate disposable saliva collection kit for rapid, reliable, and reproducible collection of saliva samples. Methods The saliva collection kit comprised of a saliva absorbent swab and an extractor unit was used to retrieve whole saliva samples from 10 subjects. The accuracy and precision of the extracted volumes (3, 10, and 30 ?l) were compared to similar volumes drawn from control samples obtained by passive drool. Additionally, the impact of kit collection method...

  17. The effect of oral and intravenous dextrose on C-peptide secretion in ponies.

    Science.gov (United States)

    de Laat, M A; van Haeften, J J; Sillence, M N

    2016-02-01

    Managing equine hyperinsulinemia is crucial for preventing laminitis, but our understanding of the mechanisms involved in insulin dysregulation in this species is incomplete. C-peptide is co-secreted with insulin but is resistant to hepatic metabolism and can be used to study insulin dysregulation. This study examined C-peptide secretion in serial blood samples collected after oral and i.v. dextrose (0.75 g/kg) administration to 9 ponies (BCS, 7.1 ± 0.5). The ponies were designated as hyperinsulinemic (HI) or normoinsulinemic (NI) responders before the study, using oral glucose tests and fasted glucose-to-insulin ratios, and responses were compared between the 2 groups. C-peptide concentrations increased ( dextrose, with similar area under the concentration-time curve (AUC) for both tests and a significant correlation with AUC. The AUC was similar in HI and NI ponies after i.v. dextrose, indicating similar pancreatic capacity for both groups. However, for oral dextrose, the AUC and the AUC were markedly higher ( < 0.05) in the HI ponies, indicating a greater secretion rate of these peptides. Slower insulin clearance might have also contributed to the larger AUC in HI ponies, but this hypothesis requires further investigation with specific measures of hepatic insulin clearance.

  18. A cGMP kinase mutant with increased sensitivity to the protein kinase inhibitor peptide PKI(5-24).

    Science.gov (United States)

    Ruth, P; Kamm, S; Nau, U; Pfeifer, A; Hofmann, F

    1996-01-01

    Synthetic peptides corresponding to the active domain of the heat-stable inhibitor protein PKI are very potent inhibitors of cAMP-dependent protein kinase, but are extremely weak inhibitors of cGMP-dependent protein kinase. In this study, we tried to confer PKI sensitivity to cGMP kinase by site-directed mutagenesis. The molecular requirements for high affinity inhibition by PKI were deduced from the crystal structure of the cAMP kinase/PKI complex. A prominent site of interaction are residues Tyr235 and Phe239 in the catalytic subunit, which from a sandwich-like structure with Phe10 of the PKI(5-24) peptide. To increase the sensitivity for PKI, the cGMP kinase codons at the corresponding sites, Ser555 and Ser559, were changed to Tyr and Phe. The mutant cGMP kinase was stimulated half maximally by cGMP at 3-fold higher concentrations (240 nM) than the wild type (77 nM). Wild type and mutant cGMP kinase did not differ significantly in their Km and Vmax for three different substrate peptides. The PKI(5-24) peptide inhibited phosphotransferase activity of the mutant cGMP kinase with higher potency than that of wild type, with Ki values of 42 +/- .3 microM and 160 +/- .7 microM, respectively. The increased affinity of the mutant cGMP kinase was specific for the PKI(5-24) peptide. Mutation of the essential Phe10 in the PKI(5-24) sequence to an Ala yielded a peptide that inhibited mutant and wild type cGMP kinase with similar potency, with Ki values of 160 +/- 11 and 169 +/- 27 microM, respectively. These results suggest that the mutations Ser555Tyr and Ser559Phe are required, but not sufficient, for high affinity inhibition of cGMP kinase by PKI.

  19. Effects of xylitol chewing gum on salivary flow rate, pH, buffering capacity and presence of Streptococcus mutans in saliva.

    Science.gov (United States)

    Ribelles Llop, M; Guinot Jimeno, F; Mayné Acién, R; Bellet Dalmau, L J

    2010-03-01

    The first studies on the use of chewing gum in dentistry were done in the 1970s. The Turku Sugar Studies, carried out between 1970 and 1973, showed the excellent anticaries properties of xylitol chewing gums. Since then, many dentists, particularly in Scandinavian countries, have studied the role of chewing xylitol-sweetened chewing gums as another preventive strategy in the control of dental caries. To compare variations in salivary flow rate, pH, buffering capacity, and levels of Streptococcus mutans in baseline conditions and after chewing paraffin pellets or xylitol chewing gum in children between the ages of 6 and 12 years who eat lunch in a school canteen. The study sample consisted of 90 children divided into 2 study groups, and a control group. The children ate lunch at the canteen of the Escultor Ortells state school in the town of Vila-real (Castellón, Spain). The baseline data recorded in the first phase of the study were compared with the data recorded in the second phase, after 15 minutes of chewing xylitol- sweetened chewing gums or paraffin pellets, depending on the study group. Salivary flow rate was measured by collecting the stimulated saliva in a graduated beaker. Levels of pH were measured using a Cyberscan pH 110 pH meter (Eutech Instruments). CRT buffer strips and the CRT bacteria test (Ivoclar-Vivadent) were used to measure buffering capacity and levels of S. mutans, respectively. The data obtained after sample collection were compared by means of a 1-way analysis of variance using the StatGraphics Plus statistical software package, version 5.0. Statistically significant differences were found (ppH, buffering capacity and levels of S. mutans were compared between the 3 groups. Comparison of salivary flow rates revealed no statistically significant differences (p>.05), though salivary flow rates were higher in the groups where gum was chewed. The effect of chewing is essential to the stimulation of salivary flow and the resulting recovery of

  20. Postprandial C-Peptide to Glucose Ratio as a Marker of β Cell Function: Implication for the Management of Type 2 Diabetes

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    Yoshifumi Saisho

    2016-05-01

    Full Text Available C-peptide is secreted from pancreatic β cells at an equimolar ratio to insulin. Since, in contrast to insulin, C-peptide is not extracted by the liver and other organs, C-peptide reflects endogenous insulin secretion more accurately than insulin. C-peptide is therefore used as a marker of β cell function. C-peptide has been mainly used to assess the presence of an insulin-dependent state for the diagnosis of type 1 diabetes. However, recent studies have revealed that β cell dysfunction is also a core deficit of type 2 diabetes, and residual β cell function is a key factor in achieving optimal glycemic control in patients with type 2 diabetes. This review summarizes the role of C-peptide, especially the postprandial C-peptide to glucose ratio which likely better reflects maximum β cell secretory capacity compared with the fasting ratio in assessing β cell function, and discusses perspectives on its clinical utility for managing glycemic control in patients with type 2 diabetes.

  1. Effect of topical application of fluoride gel NaF 2% on enzymatic and non-enzymatic antioxidant parameters of saliva.

    Science.gov (United States)

    Leite, Mariana Ferreira; Ferreira, Nayara Ferraz D'Assumpção; Shitsuka, Caleb David Willy Moreira; Lima, Amanda Martins; Masuyama, Mônica Miyuki; Sant'Anna, Giselle Rodrigues; Yamaguti, Paula Mochidome; Polotow, Tatiana G; de Barros, Marcelo Paes

    2012-06-01

    The aim of the study was to evaluate the effect of topical fluoride gel NaF 2% application on antioxidant parameters of whole saliva from children. The saliva mechanically stimulated with parafilm was collected from 25 children (6-12 years) attending the Clinic of Paediatric Dentistry of Universidade Cruzeiro do Sul, São Paulo, Brazil, before (control group) and immediately after application of neutral fluoride gel NaF 2% (fluoride-gel group), according to the Standards for Research Using Human Subjects, Resolution 196/96 of the USA National Health Council of 10/10/1996. Afterwards, pre-post ferric-reducing antioxidant power (FRAP), trolox-equivalent antioxidant capacity (TEAC), uric acid, reduced/oxidised glutathione content (GSH/GSSG) and total peroxidase activity (TPO) were evaluated in whole saliva of both groups. All non-enzymatic antioxidant parameters were augmented by fluoride-gel NaF 2% application, whereas a notable reduction (31%) of peroxidase activity was concomitantly observed in the children's saliva (p ≤ 0.05). Nevertheless, the reducing power of saliva was kept unaltered under these circumstances (p ≤ 0.05). Despite the reduced activity of peroxidase (an important antimicrobial and antioxidant enzyme), the topical fluoride gel NaF 2% favourably stimulated the release of non-enzymatic antioxidant components of saliva, sustaining the reducing power of saliva and the natural defences of the oral cavity. Copyright © 2011 Elsevier Ltd. All rights reserved.

  2. Improving Multi-Epitope Long Peptide Vaccine Potency by Using a Strategy that Enhances CD4+ T Help in BALB/c Mice.

    Directory of Open Access Journals (Sweden)

    Haniyeh Ghaffari-Nazari

    Full Text Available Peptide-based vaccines are attractive approaches for cancer immunotherapy; but the success of these vaccines in clinical trials have been limited. Our goal is to improve immune responses and anti-tumor effects against a synthetic, multi-epitope, long peptide from rat Her2/neu (rHer2/neu using the help of CD4+ T cells and appropriate adjuvant in a mouse tumor model. Female BALB/c mice were vaccinated with P5+435 multi-epitope long peptide that presents epitopes for cytotoxic T lymphocytes (CTL in combination with a universal Pan DR epitope (PADRE or CpG-oligodeoxynucleotides (CpG-ODNs as a Toll-like receptor agonist adjuvant. The results show that vaccination with the multi-epitope long peptide in combination with the PADRE peptide and CpG-ODN induced expansion of subpopulations of CD4+ and CD8+ cells producing IFN-γ, the average tumor size in the vaccinated mice was less than that of the other groups, and tumor growth was inhibited in 40% of the mice in the vaccinated group. The mean survival time was 82.6 ± 1.25 days in mice vaccinated with P5+435 + CpG+ PADRE. Our results demonstrate that inclusion of PADRE and CpG with the peptide vaccine enhanced significant tumor specific-immune responses in vaccinated mice.

  3. Polymeric pH nanosensor with extended measurement range bearing octaarginine as cell penetrating peptide

    DEFF Research Database (Denmark)

    Ke, Peng; Sun, Honghao; Liu, Mingxing

    2016-01-01

    A synthetic peptide octaarginine which mimics human immunodeficiency virus-1, Tat protein is used as cell penetrating moiety for new pH nanosensors which demonstrate enhanced cellular uptake and expanded measurement range from pH 3.9 to pH 7.3 by simultaneously incorporating two complemental pH-s......H-sensitive fluorophores in a same nanoparticle. The authors believe that this triple fluorescent pH sensor provides a new tool to pH measurements that can have application in cellular uptake mechanism study and new nanomedicine design.......A synthetic peptide octaarginine which mimics human immunodeficiency virus-1, Tat protein is used as cell penetrating moiety for new pH nanosensors which demonstrate enhanced cellular uptake and expanded measurement range from pH 3.9 to pH 7.3 by simultaneously incorporating two complemental p...

  4. Salivary pH: A diagnostic biomarker.

    Science.gov (United States)

    Baliga, Sharmila; Muglikar, Sangeeta; Kale, Rahul

    2013-07-01

    Saliva contains a variety of host defense factors. It influences calculus formation and periodontal disease. Different studies have been done to find exact correlation of salivary biomarkers with periodontal disease. With a multitude of biomarkers and complexities in their determination, the salivary pH may be tried to be used as a quick chairside test. The aim of this study was to analyze the pH of saliva and determine its relevance to the severity of periodontal disease. The study population consisted of 300 patients. They were divided into three groups of 100 patients each: Group A had clinically healthy gingiva, Group B who had generalized chronic gingivitis and Group C who had generalized chronic periodontitis. The randomized unstimulated saliva from each patient was collected and pH was tested. Data was analyzed statistically using analysis of variance technique. The salivary pH was more alkaline for patients with generalized chronic gingivitis as compared with the control group (P = 0.001) whereas patients with generalized chronic periodontitis had more acidic pH as compared with the control group (P = 0.001). These results indicate a significant change in the pH depending on the severity of the periodontal condition. The salivary pH shows significant changes and thus relevance to the severity of periodontal disease. Salivary pH may thus be used as a quick chairside diagnostic biomarker.

  5. Mensuração do potencial erosivo de balas dissolvidas em água e saliva artificial

    Directory of Open Access Journals (Sweden)

    Brenda BONVINI

    Full Text Available Resumo Introdução O consumo excessivo e frequente de balas ácidas pode estar associado à etiologia da erosão dental. Objetivo Mensurar e comparar o pH e a acidez titulável de balas dissolvidas em água e saliva artificial. Material e método Foram utilizadas as balas Tic Tac® sabores laranja, cereja/maracujá e morango, agrupadas em dois grupos: G-1: balas dissolvidas em água; G-2: balas dissolvidas em saliva artificial. As balas foram trituradas e, do pó resultante de cada sabor, foram pesadas duas amostras de 20 g, sendo então dissolvidas em 120 mL de água destilada ou 120 mL de saliva artificial. Destas soluções, obtiveram-se três amostras de 30 mL para cada um dos sabores, permitindo a leitura em triplicata do pH e da acidez titulável. O pH foi mensurado utilizando-se um potenciômetro e eletrodo combinado de vidro, previamente calibrado com soluções padrão pH 7,0 e pH 4,0. Para a verificação da acidez titulável, adicionaram-se alíquotas de 100 μL NaOH 1M, até o pH alcançar 5,5. Submeteram-se os resultados à Análise de Variância (ANOVA. As comparações das médias foram realizadas pelo Teste Tukey, em um nível de 5% de significância (p<0,05. Resultado Todas as balas apresentaram pH abaixo do crítico para dissolução do esmalte, quando dissolvidas em água e saliva artificial. Na comparação entre os grupos, o G-2, mostrou um pH mais elevado e menor acidez titulável, diferindo significantemente do G-1. Conclusão Todas as balas dissolvidas em água e saliva artificial mostraram-se potencialmente erosivas, podendo contribuir para a etiologia da erosão dental.

  6. Islet Cell Associated Autoantibodies and C-Peptide Levels in Patients with Diabetes and Symptoms of Gastroparesis

    Directory of Open Access Journals (Sweden)

    Elias S. Siraj

    2018-02-01

    Full Text Available IntroductionIndividuals with diabetes are at increased risk for complications, including gastroparesis. Type 1 diabetes mellitus (T1DM is an autoimmune disorder resulting in decreased beta-cell function. Glutamic acid decarboxylase-65 antibody (GADA is the most commonly used test to assess autoimmunity while C-peptide level is used to assess beta-cell function. Patients with type 2 diabetes mellitus (T2DM, who are GADA positive, are labeled latent autoimmune diabetes in adults (LADA.ObjectiveTo characterize patients with T1 and T2DM who have symptoms of gastroparesis using GADA and C-peptide levels and to look for association with the presence of gastroparesis and its symptom severity.Design113 T1DM and 90 T2DM patients with symptoms suggestive of gastroparesis were studied. Symptom severity was assessed using Gastroparesis Cardinal Symptom Index (GCSI. Serum samples were analyzed for GADA and C-peptide.ResultsDelayed gastric emptying was present in 91 (81% of T1DM and 60 (67% of T2DM patients (p = 0.04. GADA was present in 13% of T2DM subjects [10% in delayed gastric emptying and 20% in normal gastric emptying (p = 0.2]. Gastric retention and GCSI scores were mostly similar in GADA positive and negative T2DM patients. GADA was present in 45% of T1DM subjects [46% in delayed gastric emptying and 41% in normal gastric emptying (p = 0.81]. Low C-peptide levels were seen in 79% T1DM patients and 8% T2DM. All seven T2DM patients with low C-peptide were taking insulin compared to 52% of T2DM with normal C-peptide.ConclusionGADA was present in 13% while low C-peptide was seen in 8% of our T2DM patients with symptoms of gastroparesis. Neither did correlate with degree of delayed gastric emptying or symptom severity.ClinicalTrials.gov IdentifierNCT01696747.

  7. A Novel Apolipoprotein C-II Mimetic Peptide That Activates Lipoprotein Lipase and Decreases Serum Triglycerides in Apolipoprotein E–Knockout Mice

    Science.gov (United States)

    Sakurai, Toshihiro; Sakurai-Ikuta, Akiko; Sviridov, Denis; Freeman, Lita; Ahsan, Lusana; Remaley, Alan T.

    2015-01-01

    Apolipoprotein A-I (apoA-I) mimetic peptides are currently being developed as possible new agents for the treatment of cardiovascular disease based on their ability to promote cholesterol efflux and their other beneficial antiatherogenic properties. Many of these peptides, however, have been reported to cause transient hypertriglyceridemia due to inhibition of lipolysis by lipoprotein lipase (LPL). We describe a novel bihelical amphipathic peptide (C-II-a) that contains an amphipathic helix (18A) for binding to lipoproteins and stimulating cholesterol efflux as well as a motif based on the last helix of apolipoprotein C-II (apoC-II) that activates lipolysis by LPL. The C-II-a peptide promoted cholesterol efflux from ATP-binding cassette transporter ABCA1-transfected BHK cells similar to apoA-I mimetic peptides. Furthermore, it was shown in vitro to be comparable to the full-length apoC-II protein in activating lipolysis by LPL. When added to serum from a patient with apoC-II deficiency, it restored normal levels of LPL-induced lipolysis and also enhanced lipolysis in serum from patients with type IV and V hypertriglyceridemia. Intravenous injection of C-II-a (30 mg/kg) in apolipoprotein E–knockout mice resulted in a significant reduction of plasma cholesterol and triglycerides of 38 ± 6% and 85 ± 7%, respectively, at 4 hours. When coinjected with the 5A peptide (60 mg/kg), the C-II-a (30 mg/kg) peptide was found to completely block the hypertriglyceridemic effect of the 5A peptide in C57Bl/6 mice. In summary, C-II-a is a novel peptide based on apoC-II, which promotes cholesterol efflux and lipolysis and may therefore be useful for the treatment of apoC-II deficiency and other forms of hypertriglyceridemia. PMID:25395590

  8. Synthesis of two S-(methyl-3H)-labelled enkephalins and S-(methyl-14C) substance P

    International Nuclear Information System (INIS)

    Naegren, K.; Laangstroem, B.; Franzen, H.M.; Ragnarsson, U.

    1988-01-01

    The synthesis of 3 H-labelled Met-enkephalin and Tyr-D-Ala-Gly-Phe-Met-NH 2 (DALA) and 14 C-labelled Substance P (SP) from previously described, fully protected intermediates is reported. The labelled peptides were prepared by methylation with ( 3 H)- or ( 14 C)methyl iodide of the sulphide anions formed on deprotection of the corresponding S-benzyl-homocysteine precursors with sodium in liquid ammonia. After purification by LC, the labelled peptides were obtained in radiochemical yields in the range of 9 to 24% with a radiochemical purity higher than 97%. The specific radioactivities of the 3 H- and 14 C- labelled products, corresponding to the labelled methyl iodides used, were 80 mCi/μmol and 60 μCi/μmol, respectively. (author)

  9. Substance P receptor desensitization requires receptor activation but not phospholipase C

    International Nuclear Information System (INIS)

    Sugiya, Hiroshi; Putney, J.W. Jr.

    1988-01-01

    Previous studies have shown that exposure of parotid acinar cells to substance P at 37 degree C results in activation of phospholipase C, formation of [ 3 H]inositol 1,4,5-trisphosphate (IP 3 ), and persistent desensitization of the substance P response. In cells treated with antimycin in medium containing glucose, ATP was decreased to ∼20% of control values, IP 3 formation was completely inhibited, but desensitization was unaffected. When cells were treated with antimycin in the absence of glucose, cellular ATP was decreased to ∼5% of control values, and both IP 3 formation and desensitization were blocked. A series of substance P-related peptides increased the formation of [ 3 H]IP 3 and induced desensitization of the substance P response with a similar rank order of potencies. The substance P antagonist, [D-Pro 2 , D-Try 7,9 ]-substance P, inhibited substance P-induced IP 3 formation and desensitization but did not induce desensitization. These results suggest that the desensitization of substance P-induced IP 3 formation requires agonist activation of a P-type substance P receptor, and that one or more cellular ATP-dependent processes are required for this reaction. However, activation of phospholipase C and the generation of inositol phosphates does not seem to be a prerequisite for desensitization

  10. Antibodies to ribosomal P proteins of Trypanosoma cruzi in Chagas disease possess functional autoreactivity with heart tissue and differ from anti-P autoantibodies in lupus.

    Science.gov (United States)

    Kaplan, D; Ferrari, I; Bergami, P L; Mahler, E; Levitus, G; Chiale, P; Hoebeke, J; Van Regenmortel, M H; Levin, M J

    1997-09-16

    Anti-P antibodies present in sera from patients with chronic Chagas heart disease (cChHD) recognize peptide R13, EEEDDDMGFGLFD, which encompasses the C-terminal region of the Trypanosoma cruzi ribosomal P1 and P2 proteins. This peptide shares homology with the C-terminal region (peptide H13 EESDDDMGFGLFD) of the human ribosomal P proteins, which is in turn the target of anti-P autoantibodies in systemic lupus erythematosus (SLE), and with the acidic epitope, AESDE, of the second extracellular loop of the beta1-adrenergic receptor. Anti-P antibodies from chagasic patients showed a marked preference for recombinant parasite ribosomal P proteins and peptides, whereas anti-P autoantibodies from SLE reacted with human and parasite ribosomal P proteins and peptides to the same extent. A semi-quantitative estimation of the binding of cChHD anti-P antibodies to R13 and H13 using biosensor technology indicated that the average affinity constant was about 5 times higher for R13 than for H13. Competitive enzyme immunoassays demonstrated that cChHD anti-P antibodies bind to the acidic portions of peptide H13, as well as to peptide H26R, encompassing the second extracellular loop of the beta1 adrenoreceptor. Anti-P antibodies isolated from cChHD patients exert a positive chronotropic effect in vitro on cardiomyocytes from neonatal rats, which resembles closely that of anti-beta1 receptor antibodies isolated from the same patient. In contrast, SLE anti-P autoantibodies have no functional effect. Our results suggest that the adrenergic-stimulating activity of anti-P antibodies may be implicated in the induction of functional myocardial impairments observed in cChHD.

  11. A simple high-performance liquid chromatography for the determination of linezolid in human plasma and saliva.

    Science.gov (United States)

    Hara, Shuuji; Uchiyama, Masanobu; Yoshinari, Masami; Matsumoto, Taichi; Jimi, Shiro; Togawa, Atsushi; Takata, Tohru; Takamatsu, Yasushi

    2015-09-01

    Linezolid is an antimicrobial agent for the treatment of multiresistant Gram-positive infections. A practical high-performance liquid chromatography method was developed for the determination of linezolid in human plasma and saliva. Linezolid and an internal standard (o-ethoxybenzamide) were extracted from plasma and saliva with ethyl acetate and analyzed on a Capcell Pak C18 MG column with UV detection at 254 nm. The calibration curve was linear through the range 0.5-50 µg/mL using a 200 μL sample volume. The intra- and interday precisions were all plasma and 5.60% for saliva. The accuracies ranged from 98.8 to 110% for both matrices. The mean recoveries of linezolid were 80.8% for plasma and 79.0% for saliva. This method was used to determine the plasma and saliva concentrations of linezolid in healthy volunteers who were orally administered a 600 mg dose of linezolid. Our liquid-liquid extraction procedure is easy and requires a small volume of plasma or saliva (200 μL). This small volume can be advantageous in clinical pharmacokinetic studies, especially if children participate. Copyright © 2015 John Wiley & Sons, Ltd.

  12. Biochemical Characterisation of Phage Pseudomurein Endoisopeptidases PeiW and PeiP Using Synthetic Peptides

    OpenAIRE

    Schofield, Linley R.; Beattie, Amy K.; Tootill, Catherine M.; Dey, Debjit; Ronimus, Ron S.

    2015-01-01

    Pseudomurein endoisopeptidases cause lysis of the cell walls of methanogens by cleaving the isopeptide bond Ala-ε-Lys in the peptide chain of pseudomurein. PeiW and PeiP are two thermostable pseudomurein endoisopeptidases encoded by phage ΨM100 of Methanothermobacter wolfei and phages ΨM1 and ΨM2 of Methanothermobacter marburgensis, respectively. A continuous assay using synthetic peptide substrates was developed and used in the biochemical characterisation of recombinant PeiW and PeiP. The a...

  13. Hubungan Body Mass Index dengan Laju Aliran Saliva (Studi pada Mahasiswa Fakultas Kedokteran Gigi Universitas Andalas

    Directory of Open Access Journals (Sweden)

    Fildzah Nurul Fajrin

    2015-12-01

    Relationship between Body Mass Index and Saliva Flow Rate (A Study in Faculty of Dentistry, Andalas University. Saliva flow rate is an affecting factor of caries formation. Adequate saliva flow rate can prevent cariess formation process and progression of periodontal disease and oral infection. Nutritional status is known as an influencing factor of saliva flow rate. The parameter of nutritional status is Body Mass Index (BMI. The purpose of this study is to discover the relation between BMI and saliva flow rate in College Students of the Dentistry Faculty Andalas University. This observational study with cross sectional design was conducted to 24 college students of the Dentistry Faculty Andalas University whose ages were 18-25 years old who complied with inclussion and exclussion criteria. BMI was calculated with BMI formula. The saliva flow rate was determined with Sialometry Method of Navazesh 2008 with ”ml/minute” unit. The univariat data analysis was caried out to describe each variable. Normality test Kolmogorov Smirnoff was done to see the normal distribution (p > 0,05. After the distribution was proven to be normal, corelation and regression test was executed to discover the relation between BMI and saliva flow rate. Based on the correlation test result, BMI of college students of the Dentistry Faculty Andalas University is χχ ± SD = 24,6 ± 6,02 kg/m2 and saliva flow rate is χχ ± SD = 0,29 ± 0,1 ml/minute. The relation between BMI and saliva flow rate expresses medium correlation with negative direction ( r = - 0,451. The result of linear regression shows that Saliva Flow Rate = 0,404 - 0,008*(BMI. The coefficient of regression BMI 0.008 shows that increasing 1 unit of BMI will decrease saliva flow rate 0,008 ml/minute. This study concludes that there is correlation between Body Mass Index and saliva flow rate. Obesity group has the lowest saliva flow rate, while the saliva flow rate does not decrease in underweight group.

  14. Meta-genome-wide association studies identify a locus on chromosome 1 and multiple variants in the MHC region for serum C-peptide in type 1 diabetes.

    Science.gov (United States)

    Roshandel, Delnaz; Gubitosi-Klug, Rose; Bull, Shelley B; Canty, Angelo J; Pezzolesi, Marcus G; King, George L; Keenan, Hillary A; Snell-Bergeon, Janet K; Maahs, David M; Klein, Ronald; Klein, Barbara E K; Orchard, Trevor J; Costacou, Tina; Weedon, Michael N; Oram, Richard A; Paterson, Andrew D

    2018-05-01

    The aim of this study was to identify genetic variants associated with beta cell function in type 1 diabetes, as measured by serum C-peptide levels, through meta-genome-wide association studies (meta-GWAS). We performed a meta-GWAS to combine the results from five studies in type 1 diabetes with cross-sectionally measured stimulated, fasting or random C-peptide levels, including 3479 European participants. The p values across studies were combined, taking into account sample size and direction of effect. We also performed separate meta-GWAS for stimulated (n = 1303), fasting (n = 2019) and random (n = 1497) C-peptide levels. In the meta-GWAS for stimulated/fasting/random C-peptide levels, a SNP on chromosome 1, rs559047 (Chr1:238753916, T>A, minor allele frequency [MAF] 0.24-0.26), was associated with C-peptide (p = 4.13 × 10 -8 ), meeting the genome-wide significance threshold (p C>T, MAF 0.07-0.10, p = 8.43 × 10 -8 ). In the stimulated C-peptide meta-GWAS, rs61211515 (Chr6:30100975, T/-, MAF 0.17-0.19) in the MHC region was associated with stimulated C-peptide (β [SE] = - 0.39 [0.07], p = 9.72 × 10 -8 ). rs61211515 was also associated with the rate of stimulated C-peptide decline over time in a subset of individuals (n = 258) with annual repeated measures for up to 6 years (p = 0.02). In the meta-GWAS of random C-peptide, another MHC region, SNP rs3135002 (Chr6:32668439, C>A, MAF 0.02-0.06), was associated with C-peptide (p = 3.49 × 10 -8 ). Conditional analyses suggested that the three identified variants in the MHC region were independent of each other. rs9260151 and rs3135002 have been associated with type 1 diabetes, whereas rs559047 and rs61211515 have not been associated with a risk of developing type 1 diabetes. We identified a locus on chromosome 1 and multiple variants in the MHC region, at least some of which were distinct from type 1 diabetes risk loci, that were associated with C-peptide

  15. Analysis of risperidone and 9-hydroxyrisperidone in human plasma, urine and saliva by MEPS-LC-UV.

    Science.gov (United States)

    Mandrioli, Roberto; Mercolini, Laura; Lateana, Domenico; Boncompagni, Giancarlo; Raggi, Maria Augusta

    2011-01-15

    Risperidone is currently one of the most frequently prescribed atypical antipsychotic drugs; its main active metabolite 9-hydroxyrisperidone contributes significantly to the therapeutic effects observed. An original analytical method is presented for the simultaneous analysis of risperidone and the metabolite in plasma, urine and saliva by high-performance liquid chromatography coupled to an original sample pre-treatment procedure based on micro-extraction by packed sorbent (MEPS). The assays were carried out using a C8 reversed-phase column and a mobile phase composed of 73% (v/v) acidic phosphate buffer (30 mM, pH 3.0) containing 0.23% triethylamine and 27% (v/v) acetonitrile. The UV detector was set at 238 nm and diphenhydramine was used as the internal standard. The sample pre-treatment by MEPS was carried out on a C8 sorbent. The extraction yields values were higher than 92% for risperidone and 90% for 9-hydroxyrisperidone, with RSD for precision always lower than 7.9% for both analytes. Limit of quantification values in the different matrices were 4 ng/mL or lower for risperidone and 6 ng/mL or lower for the metabolite. The method was successfully applied to plasma, urine and saliva samples from psychotic patients undergoing therapy with risperidone, with satisfactory accuracy results (recovery>89%) and no interference from other drugs. Thus, the method seems to be suitable for the therapeutic drug monitoring of schizophrenic patients using the three different biological matrices plasma, urine and saliva. Copyright © 2010 Elsevier B.V. All rights reserved.

  16. Saliva: Physiology and Diagnostic Potential in Health and Disease

    Directory of Open Access Journals (Sweden)

    Sebastien J. C. Farnaud

    2010-01-01

    Full Text Available Saliva has been described as the mirror of the body. In a world of soaring healthcare costs and an environment where rapid diagnosis may be critical to a positive patient outcome, saliva is emerging as a viable alternative to blood sampling. In this review, we discuss the composition and various physiological roles of saliva in the oral cavity, including soft tissue protection, antimicrobial activities, and oral tissue repair. We then explore saliva as a diagnostic marker of local oral disease and focus particularly on oral cancers. The cancer theme continues when we focus on systemic disease diagnosis from salivary biomarkers. Communicable disease is the focus of the next section where we review the literature relating to the direct and indirect detection of pathogenic infections from human saliva. Finally, we discuss hormones involved in appetite regulation and whether saliva is a viable alternative to blood in order to monitor hormones that are involved in satiety.

  17. Recombinant heat shock protein 70 functional peptide and alpha-fetoprotein epitope peptide vaccine elicits specific anti-tumor immunity.

    Science.gov (United States)

    Wang, Xiao-Ping; Wang, Qiao-Xia; Lin, Huan-Ping; Xu, Bing; Zhao, Qian; Chen, Kun

    2016-11-01

    Alpha-fetoprotein (AFP) is a marker of hepatocellular carcinoma (HCC) and serves as a target for immunotherapy. However, current treatments targeting AFP are not reproducible and do not provide complete protection against cancer. This issue may be solved by developing novel therapeutic vaccines with enhanced immunogenicity that could effectively target AFP-expressing tumors. In this study, we report construction of a therapeutic peptide vaccine by linking heat shock protein 70 (HSP70) functional peptide to the AFP epitope to obtain HSP70-P/AFP-P. This novel peptide was administered into BALB/c mice to observe the effects. Quantification of AFP-specific CD8 + T cells that secrete IFN-γ in these mice via ELISPOT revealed the synergistic effects of HSP70-P/AFP-P with increased numbers of AFP-specific CD8 + T cells. Similarly, ELISA analysis showed increased granzyme B and perforin released by natural killer cells. Moreover, in vitro cytotoxic T-lymphocyte assays and in vivo tumor preventive experiments clearly showed the higher antitumor effects of HSP70-P/AFP-P against AFP-expressing tumors. These results show that treatment of BALB/c mice with HSP70-P/AFP-P induced stronger T-cells responses and improved protective immunity. Our data suggest that HSP70-P/AFP-P may be used as a therapeutic approach in the treatment of AFP-expressing cancers.

  18. Saliva composition in three selected groups with normal stimulated salivary flow rates, but yet major differences in caries experience and dental erosion.

    Science.gov (United States)

    Bardow, Allan; Lykkeaa, Joan; Qvist, Vibeke; Ekstrand, Kim; Twetman, Svante; Fiehn, Niels-Erik

    2014-08-01

    It was hypothesized that, by comparing matched subjects with major differences in these dental diseases, but yet normal saliva flow rates, it would be possible to obtain data on the effect of saliva composition on dental disease isolated from the effect of the flow rate. Thus, the aim of the study was to compare the major physicochemical characteristics of stimulated whole saliva in three groups of 85 subjects, each with normal saliva flow rates and at least 24 remaining teeth. A group with very little dental disease (healthy), a group with dental erosion (erosion) and a group with very high caries experience (caries) were chosen. Furthermore, the aim was to determine whether differences among groups could also be found on an individual level. Although it was not possible to retrieve three groups whose members were completely identical, the present study points in the direction that, on a group level, subjects with very little dental disease seemed to have a more favorable physicochemical saliva composition with respect to higher calcium, phosphate, bicarbonate, pH, degree of saturation with respect to hydroxyapatite and a lower critical pH (p dental erosion (p dental caries and erosion in single individuals.

  19. C-Peptide Decline in Type 1 Diabetes Has Two Phases: An Initial Exponential Fall and a Subsequent Stable Phase.

    Science.gov (United States)

    Shields, Beverley M; McDonald, Timothy J; Oram, Richard; Hill, Anita; Hudson, Michelle; Leete, Pia; Pearson, Ewan R; Richardson, Sarah J; Morgan, Noel G; Hattersley, Andrew T

    2018-06-07

    The decline in C-peptide in the 5 years after diagnosis of type 1 diabetes has been well studied, but little is known about the longer-term trajectory. We aimed to examine the association between log-transformed C-peptide levels and the duration of diabetes up to 40 years after diagnosis. We assessed the pattern of association between urinary C-peptide/creatinine ratio (UCPCR) and duration of diabetes in cross-sectional data from 1,549 individuals with type 1 diabetes using nonlinear regression approaches. Findings were replicated in longitudinal follow-up data for both UCPCR ( n = 161 individuals, 326 observations) and plasma C-peptide ( n = 93 individuals, 473 observations). We identified two clear phases of C-peptide decline: an initial exponential fall over 7 years (47% decrease/year [95% CI -51%, -43%]) followed by a stable period thereafter (+0.07%/year [-1.3, +1.5]). The two phases had similar durations and slopes in patients above and below the median age at diagnosis (10.8 years), although levels were lower in the younger patients irrespective of duration. Patterns were consistent in both longitudinal UCPCR ( n = 162; ≤7 years duration: -48%/year [-55%, -38%]; >7 years duration -0.1% [-4.1%, +3.9%]) and plasma C-peptide ( n = 93; >7 years duration only: -2.6% [-6.7%, +1.5%]). These data support two clear phases of C-peptide decline: an initial exponential fall over a 7-year period, followed by a prolonged stabilization where C-peptide levels no longer decline. Understanding the pathophysiological and immunological differences between these two phases will give crucial insights into understanding β-cell survival. © 2018 by the American Diabetes Association.

  20. Nanovesicle encapsulation of antimicrobial peptide P34: physicochemical characterization and mode of action on Listeria monocytogenes

    Science.gov (United States)

    da Silva Malheiros, Patrícia; Sant'Anna, Voltaire; Micheletto, Yasmine Miguel Serafini; da Silveira, Nadya Pesce; Brandelli, Adriano

    2011-08-01

    Antimicrobial peptide P34, a substance showing antibacterial activity against pathogenic and food spoilage bacteria, was encapsulated in liposomes prepared from partially purified soybean phosphatidylcholine, and their physicochemical characteristics were evaluated. The antimicrobial activity was estimated by agar diffusion assay using Listeria monocytogenes ATCC 7644 as indicator strain. A concentration of 3,200 AU/mL of P34 was encapsulated in nanovesicles and stocked at 4 °C. No significant difference ( p > 0.05) in the biological activity of free and encapsulated P34 was observed through 24 days. Size and PDI of liposomes, investigated by light scattering analysis, were on average 150 nm and 0.22 respectively. Zeta potential was -27.42 mV. There was no significant change ( p > 0.05) in the physicochemical properties of liposomes during the time of evaluation. The liposomes presented closed spherical morphology as visualized by transmission electron microscopy (TEM). The mode of action of liposome-encapsulated P34 under L. monocytogenes cells was investigated by TEM. Liposomes appeared to adhere but not fuse with the bacterial cell wall, suggesting that the antimicrobial is released from nanovesicles to act against the microorganism. The effect of free and encapsulated P34 was tested against L. monocytogenes, showing that free bacteriocin inhibited the pathogen more quickly than the encapsulated P34. Liposomes prepared with low-cost lipid showed high encapsulation efficiency for a new antimicrobial peptide and were stable during storage. The mode of action against the pathogen L. monocytogenes was characterized.

  1. Computational Strategy for Quantifying Human Pesticide Exposure based upon a Saliva Measurement

    Directory of Open Access Journals (Sweden)

    Charles eTimchalk

    2015-05-01

    Full Text Available Quantitative exposure data is important for evaluating toxicity risk and biomonitoring is a critical tool for evaluating human exposure. Direct personal monitoring provides the most accurate estimation of a subject’s true dose, and non-invasive methods are advocated for quantifying exposure to xenobiotics. In this regard, there is a need to identify chemicals that are cleared in saliva at concentrations that can be quantified to support the implementation of this approach. This manuscript reviews the computational modeling approaches that are coupled to in vivo and in vitro experiments to predict salivary uptake and clearance of xenobiotics and provides additional insight on species-dependent differences in partitioning that are of key importance for extrapolation. The primary mechanism by which xenobiotics leave the blood and enter saliva involves paracellular transport, passive transcellular diffusion, or trancellular active transport with the majority of xenobiotics transferred by passive diffusion. The transcellular or paracellular diffusion of unbound chemicals in plasma to saliva has been computationally modeled using compartmental and physiologically based approaches. Of key importance for determining the plasma:saliva partitioning was the utilization of the Schmitt algorithm that calculates partitioning based upon the tissue composition, pH, chemical pKa and plasma protein-binding. Sensitivity analysis identified that both protein-binding and pKa (for weak acids and bases have significant impact on determining partitioning and species dependent differences based upon physiological variance. Future strategies are focused on an in vitro salivary acinar cell based system to experimentally determine and computationally predict salivary gland uptake and clearance for xenobiotics. It is envisioned that a combination of salivary biomonitoring and computational modeling will enable the non-invasive measurement of chemical exposures in human

  2. IN VITRO Cr(VI SPECIATION IN SYNTHETIC SALIVA AFTER RELEASING FROM ORTHODONTIC BRACKETS USING SILICA-APTES SEPARATION AND GF AAS DETERMINATION

    Directory of Open Access Journals (Sweden)

    Maciel S. Luz

    Full Text Available A method for Cr(VI speciation in synthetic saliva after releasing from orthodontic brackets, using silica nanoparticles organofunctionalized with (3-aminopropyltriethoxysilane (APTES for Cr(III/Cr(VI separation and GF AAS determination is proposed. Under the optimized conditions, Cr(VI speciation was performed using 150 mg of silica organofunctionalized with 2.0% (v v-1 of APTES at pH 8. It was observed different sensitivity when calibrations of GF AAS were performed using Cr(III or Cr(VI as standard solutions. Consequently, calibrations using stoichiometric mixtures (Cr(III + Cr(VI were used for total Cr determination and calibration using Cr(VI was used only for the determination of this specie. The reliability of the proposed silica-APTES separation procedure and GF AAS determination was checked by addition of both species in synthetic saliva. Recoveries ranging from 97 to 110% were obtained. The repeatability, based on the relative standard deviation (RSD inter days was less than 6%. A corrosion test was carried out on 20 orthodontic brackets from two different models, after immersion in synthetic saliva (pH=6.0 at 37 °C with agitation (125 rpm for 24 h. It was observed that about 40% of the total chromium released from the analyzed orthodontic brackets was Cr(VI.

  3. Artificial saliva effect on toxic substances release from acrylic resins

    Directory of Open Access Journals (Sweden)

    Kostić Milena

    2015-01-01

    Full Text Available Background/Aim. Acrylic-based resins are intensively used in dentistry practice as restorative or denture-base materials. The purpose of this study was to analyze the surface structure of denture base resins and the amount of released potentially toxic substances (PTS immediately upon polymerization and incubation in different types of artificial saliva. Methods. Storage of acrylic samples in two models of artificial saliva were performed in a water bath at the temperature of 37 ± 1°C. Analysis of the surface structure of samples was carried out using scanning electronic microscopy analysis immediately after polymerization and after the 30-day incubation. The amounts of PTS per day, week and month extracts were measured using high-pressure liquid chromatography. Results. Surface design and amount of PTS in acrylic materials were different and depended on the types and duration of polymerization. The surfaces of tested acrylates became flatter after immersing in solutions of artificial saliva. The degree of acrylic materials release was not dependent on the applied model of artificial saliva. Conclusion. In order to improve biological features of acrylic resin materials, it was recommended that dentures lined with soft or hard coldpolymerized acrylates should be kept at least 1 to 7 days in water before being given to a patient. So, as to reach high degree of biocompatibility preparation of prosthetic restorations from heat-polymerized acrylate was unnecessary. [Projekat Ministarstva nauke Republike Srbije, br. 41017

  4. HEPATITIS B VIRUS DNA IN SALIVA FROM CHILDREN WITH CHRONIC HEPATITIS B INFECTION IMPLICATIONS FOR SALIVA AS A POTENTIAL MODE OF HORIZONTAL TRANSMISSION

    NARCIS (Netherlands)

    Heiberg, Ida Louise; Hoegh, Mette; Ladelund, Steen; Niesters, Hubert G. M.; Hogh, Birthe

    2010-01-01

    To explore the mechanism of horizontal transmission of hepatitis B virus (HBV) among children, we investigated the quantitative relationship between HBV in saliva and blood from 46 children with chronic hepatitis B. We found high levels of HBV DNA in saliva of HBeAg (+) children, suggesting saliva

  5. Hepatitis B virus DNA in saliva from children with chronic hepatitis B infection: implications for saliva as a potential mode of horizontal transmission

    DEFF Research Database (Denmark)

    Heiberg, Ida Louise; Hoegh, Mette; Ladelund, Steen

    2010-01-01

    To explore the mechanism of horizontal transmission of hepatitis B virus (HBV) among children, we investigated the quantitative relationship between HBV in saliva and blood from 46 children with chronic hepatitis B. We found high levels of HBV DNA in saliva of HBeAg (+) children, suggesting saliva...

  6. Daily and annually variation of unstimulated whole saliva flow rate and pH and their relation with body profile in healthy young adults.

    Science.gov (United States)

    Foglio-Bonda, P L; Migliario, M; Rocchetti, V; Pattarino, F; Foglio-Bonda, A

    2013-09-01

    To analyse pH and flow rate (FR) of unstimulated whole saliva (UWS), detecting their possible correlations both among themselves and with body profile; in addition to identify daily, annually and gender differences. Eighty-one (47 ♀; 34 ♂) healthy young adults (mean age 22.7±4.09 years old) were enrolled. Saliva was sampled using spitting method. The data were statistically analysed using Pearson's coefficient, ANOVA or Kruskal-Wallis test, Student's t test or the Wilcoxon-Mann-Whitney test. The mean UWS/FR was 0.643 ml/min (range 0.164-1.656 ml/min; percentile 25 = 0.400 ml/min; percentile 50 = 0.643 ml/min, percentile 75 = 0.832 ml/min; median = 0.590 ml/min) and no significant differences were found in gender. The mean UWS/pH was 6.95 (range 6.06-7.91, S.D. 0.28, RSD % 4.08): pH was higher in males (7.02) than females (6.92; p = 0.009). The UWS/FR increased almost steadily during the day: from 0.593 ml/min at 9:00 to 0.669 ml/min at 17:00 (p = 0.04), the greatest increase was found between 9:00 and 11:00. Through the seasons the UWS/FR decreased from summer to spring with a difference of 0.048 ml/min (p pH showed a slight increase between 9:00 and 17:00 (p pH among the seasons (max. 0.09; p pH was found (R = 0.20; p = 0.008). We did not find correlations between body profile vs UWS/FR or pH. UWS/FR varies more widely than UWS/pH: maintaining a proper acid/base balance is an essential factor for the homeostasis of the oral cavity and probably this would explain the reason for the lack of the variables evaluated influencing UWS/pH.

  7. Truncated Glucagon-like Peptide-1 and Exendin-4 α-Conotoxin pl14a Peptide Chimeras Maintain Potency and α-Helicity and Reveal Interactions Vital for cAMP Signaling in Vitro*

    Science.gov (United States)

    Swedberg, Joakim E.; Schroeder, Christina I.; Mitchell, Justin M.; Fairlie, David P.; Edmonds, David J.; Griffith, David A.; Ruggeri, Roger B.; Derksen, David R.; Loria, Paula M.; Price, David A.; Liras, Spiros; Craik, David J.

    2016-01-01

    Glucagon-like peptide-1 (GLP-1) signaling through the glucagon-like peptide 1 receptor (GLP-1R) is a key regulator of normal glucose metabolism, and exogenous GLP-1R agonist therapy is a promising avenue for the treatment of type 2 diabetes mellitus. To date, the development of therapeutic GLP-1R agonists has focused on producing drugs with an extended serum half-life. This has been achieved by engineering synthetic analogs of GLP-1 or the more stable exogenous GLP-1R agonist exendin-4 (Ex-4). These synthetic peptide hormones share the overall structure of GLP-1 and Ex-4, with a C-terminal helical segment and a flexible N-terminal tail. Although numerous studies have investigated the molecular determinants underpinning GLP-1 and Ex-4 binding and signaling through the GLP-1R, these have primarily focused on the length and composition of the N-terminal tail or on how to modulate the helicity of the full-length peptides. Here, we investigate the effect of C-terminal truncation in GLP-1 and Ex-4 on the cAMP pathway. To ensure helical C-terminal regions in the truncated peptides, we produced a series of chimeric peptides combining the N-terminal portion of GLP-1 or Ex-4 and the C-terminal segment of the helix-promoting peptide α-conotoxin pl14a. The helicity and structures of the chimeric peptides were confirmed using circular dichroism and NMR, respectively. We found no direct correlation between the fractional helicity and potency in signaling via the cAMP pathway. Rather, the most important feature for efficient receptor binding and signaling was the C-terminal helical segment (residues 22–27) directing the binding of Phe22 into a hydrophobic pocket on the GLP-1R. PMID:27226591

  8. Characterisation of human saliva as a platform for oral dissolution medium development.

    Science.gov (United States)

    Gittings, Sally; Turnbull, Neil; Henry, Brian; Roberts, Clive J; Gershkovich, Pavel

    2015-04-01

    Human saliva is a biological fluid of great importance in the field of dissolution testing. However, until now, no consensus has been reached on its key characteristics relevant to dissolution testing. As a result, it is difficult to select or develop an in vitro dissolution medium to best represent human saliva. In this study, the pH, buffer capacity, surface tension, viscosity and flow rate of both unstimulated (US) and stimulated (SS) human saliva were investigated in order to provide a platform of reference for future dissolution studies using simulated salivary fluids. Age and gender related differences in a sample size of 30 participants for each parameter were investigated. Significant differences were established between US and SS for all characteristics except surface tension. Therefore, the requirement for using two simulated salivary fluids should be considered when developing an oral dissolution model. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Peptide-DNA conjugates as tailored bivalent binders of the oncoprotein c-Jun.

    Science.gov (United States)

    Pazos, Elena; Portela, Cecilia; Penas, Cristina; Vázquez, M Eugenio; Mascareñas, José L

    2015-05-21

    We describe a ds-oligonucleotide-peptide conjugate that is able to efficiently dismount preformed DNA complexes of the bZIP regions of oncoproteins c-Fos and c-Jun (AP-1), and therefore might be useful as disrupters of AP-1-mediated gene expression pathways.

  10. Sealing effectiveness of fissure sealant bonded with universal adhesive systems on saliva-contaminated and noncontaminated enamel.

    Science.gov (United States)

    Memarpour, Mahtab; Shafiei, Fereshteh; Zarean, Mehran; Razmjoei, Faranak

    2018-01-01

    The effectiveness of sealants is dependent upon their adhesion to enamel surface. The aim of the study was to evaluate the sealing ability of a pit and fissure sealant used with a universal adhesive (etch-and-rinse vs. self-etch modes) when the site is contaminated with saliva. Adhesive properties were evaluated as microleakage and scanning electron microscopic (SEM) characteristics. A total of 72 mandibular third molars were randomly divided into 6 groups (n=12). Occlusal pits and fissures were sealed with an unfilled resin fissure sealant (FS) material with or without saliva contamination. The groups included: 1) phosphoric acid etching + FS (control), 2) phosphoric acid etching + Scotchbond Universal (etch-and-rinse) + FS, 3) phosphoric acid etching + saliva + Scotchbond Universal (etch-and-rinse) + FS, 4) Scotchbond Universal (self-etching) + FS,5) Scotchbond Universal (self-etching) + saliva + FS, and 6) Scotchbond Universal (self-etching) + saliva + Scotchbond Universal + FS. After thermocycling, the teeth were placed in 0.5% fuchsin, sectioned, and evaluated by digital microscopy. Two samples from each group were also observed by SEM. The data were analyzed with Kruskal-Wallis and Mann-Whitney tests for a significance of p universal adhesive in etch-and-rinse or self-etch modes was similar to that of conventional acid etching. Saliva contamination had a negative effect on sealant adhesion to pretreated enamel. Key words: Pit and fissure sealant, Universal adhesive, Saliva.

  11. Assessment of extracellular dehydration using saliva osmolality.

    Science.gov (United States)

    Ely, Brett R; Cheuvront, Samuel N; Kenefick, Robert W; Spitz, Marissa G; Heavens, Kristen R; Walsh, Neil P; Sawka, Michael N

    2014-01-01

    When substantial solute losses accompany body water an isotonic hypovolemia (extracellular dehydration) results. The potential for using blood or urine to assess extracellular dehydration is generally poor, but saliva is not a simple ultra-filtrate of plasma and the autonomic regulation of salivary gland function suggests the possibility that saliva osmolality (Sosm) may afford detection of extracellular dehydration via the influence of volume-mediated factors. This study aimed to evaluate the assessment of extracellular dehydration using Sosm. In addition, two common saliva collection methods and their effects on Sosm were compared. Blood, urine, and saliva samples were collected in 24 healthy volunteers during paired euhydration and dehydration trials. Furosemide administration and 12 h fluid restriction were used to produce extracellular dehydration. Expectoration and salivette collection methods were compared in a separate group of eight euhydrated volunteers. All comparisons were made using paired t-tests. The diagnostic potential of body fluids was additionally evaluated. Dehydration (3.1 ± 0.5% loss of body mass) decreased PV (-0.49 ± 0.12 L; -15.12 ± 3.94% change), but Sosm changes were marginal ( 0.05). Extracelluar dehydration was not detectable using plasma, urine, or saliva measures. Salivette and expectoration sampling methods produced similar, consistent results for Sosm, suggesting no methodological influence on Sosm.

  12. Shear Bond Strength of Orthodontic Brackets and Disinclusion Buttons: Effect of Water and Saliva Contamination

    OpenAIRE

    Sfondrini, Maria Francesca; Fraticelli, Danilo; Gandini, Paola; Scribante, Andrea

    2013-01-01

    Purpose. The aim of this study was to assess the effect of water and saliva contamination on the shear bond strength and failure site of orthodontic brackets and lingual buttons. Materials and Methods. 120 bovine permanent mandibular incisors were randomly divided into 6 groups of 20 specimens each. Both orthodontic brackets and disinclusion buttons were tested under three different enamel surface conditions: (a) dry, (b) water contamination, and (c) saliva contamination. Brackets and buttons...

  13. DNA methylation analysis from saliva samples for epidemiological studies.

    Science.gov (United States)

    Nishitani, Shota; Parets, Sasha E; Haas, Brian W; Smith, Alicia K

    2018-06-18

    Saliva is a non-invasive, easily accessible tissue, which is regularly collected in large epidemiological studies to examine genetic questions. Recently, it is becoming more common to use saliva to assess DNA methylation. However, DNA extracted from saliva is a mixture of both bacterial and human DNA derived from epithelial and immune cells in the mouth. Thus, there are unique challenges to using salivary DNA in methylation studies that can influence data quality. This study assesses: (1) quantification of human DNA after extraction; (2) delineation of human and bacterial DNA; (3) bisulfite conversion (BSC); (4) quantification of BSC DNA; (5) PCR amplification of BSC DNA from saliva and; (6) quantitation of DNA methylation with a targeted assay. The framework proposed will allow saliva samples to be more widely used in targeted epigenetic studies.

  14. Extra virgin olive oil aroma release after interaction with human saliva from individuals with different body mass index.

    Science.gov (United States)

    Genovese, Alessandro; Rispoli, Tiziana; Sacchi, Raffaele

    2018-07-01

    The interindividual variability observed in saliva characteristics raises the question of its relationship with variability in fat sensory perception, particularly in aroma compounds. In the present study, which aimed to measure aroma release from different individuals, eleven key aroma compounds of extra virgin olive oil (EVOO) were monitored and quantified in dynamic headspace after an in vitro interaction between EVOO and human saliva. Therefore, 60 individuals were studied from those who were normal weight (NW), overweight (OW) and obese (O). OW and O demonstrate a higher release of C 6 compounds compared to NW. By contrast, NW have a higher release of C 5 compounds. Pentanal and hexanal also increased after saliva interaction in a refined olive oil that is free from volatiles. Among the saliva samples with a higher release in NW individuals, only pentanal was different. However, the low levels of these lipid oxidation end-products do not appear to be very important with respect to increasing odorous fat sensitivity. The results obtained in the present study demonstrate the important role of saliva in the aroma release of EVOO, indicating that humans can perceive it differently in relation to their body mass index. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  15. Radioimmunological analysis of circadian rhythms of cortisol and melatonin in saliva

    International Nuclear Information System (INIS)

    Demel, A.W.

    1990-12-01

    Since blood cortisol (F) and melatonin (MTL) display a circadian secretion pattern and since the saliva concentration of these hormones is an excellent indicator of its blood levels the measurement of salivary F and MLT may be used for examining circadian rhythmicity. In these study the relationship between salivary F and MLT was explored. For this purpose it was necessary first to establish and validate a radioimmunoassay for F in saliva: salivary F was determined by a direct radioimmunoassay using cortisol-3-(O-carboxymethyl) oximino-(2- ( 125 I)iodohistamin) as tracer and cortisol-3-CMO-BSA antiserum. The parallel measurement of F levels in saliva and serum of adults gave an excellent correlation (r=0.87, p 0.00956x ). Serum F was assayed on the Abott TDX-System using a radioimmunofluorescence method. Secondly, using this assay the circadian saliva F pattern was determined as well as the pattern of salivary MLT in 9 young, healthy volunteers. For saliva MLT estimations a previously published method was applied (SCHULZ et al 1990). Using a computerized program (RHYTHM) written by EVE v. CAUTER (1979), the hormone data of each individuum were examined for circadian rhythmicity and its acrophases (time of occurrence of the maximum of a sinusoid fitted to the data). The F acrophase occured between 7:00 and 12:00 h (Mean: 8:06 h, SD: 93.8 min). The acrophase for MLT was found between 0:00 and 6:00 h (Mean: 3:33 h, SD: 104.4 min). There was a phase difference between both hormones of 3 to 7 hours (Mean: 292.5 min, SD: 74.7 min). The easy stress-free non-invasive nature of saliva collection makes saliva to one of the most accessible body fluids and of high value in studying the circadian system in healthy humans as well as in infants, children, pregnant women and anaemic patients. Measurements of salivary F and MLT may help to elucidate not only the circadian rhythms of these hormones under normal and pathological conditions but it may also provide insight in

  16. Binding of corroded ions to human saliva.

    Science.gov (United States)

    Mueller, H J

    1985-05-01

    Employing equilibrium dialysis, the binding abilities of Cu, Al, Co and Cr ions from corroded Cu-Al and Co-Cr dental casting alloys towards human saliva and two of its gel chromatographic fractions were determined. Results indicate that both Cu and Co bind to human saliva i.e. 0.045 and 0.027 mg/mg protein, respectively. Besides possessing the largest binding ability, Cu also possessed the largest binding capacity. The saturation of Cu binding was not reached up to the limit of 0.35 mg protein/ml employed in the tests, while Co reached full saturation at about 0.2 mg protein/ml. Chromium showed absolutely no binding to human saliva while Al ions did not pass through the dialysis membranes. Compared to the binding with solutions that were synthetically made up to contain added salivary-type proteins, it is shown that the binding to human saliva is about 1 order of magnitude larger, at least for Cu ions.

  17. Study of experiment on leaching of bisphenol A from infant books to artificial saliva.

    Science.gov (United States)

    Sajiki, Junko; Yanagibori, Ryoko; Kobayashi, Yaeko

    2010-05-01

    To assess the risk of bisphenol A (BPA) exposure when infants suck or chew infant books, the concentration of BPA leaching from infant books published by Japanese makers to artificial saliva was measured. The concentration of BPA leaching from 10 infant books to 15 ml artificial saliva or water was measured at 37 degrees C for 20 hrs. BPA concentration was measured by high-performance liquid chromatography-electrochemical detection (HPLC-ECD) with solid-phase extraction. BPA was leached from all books when pieces of them were dipped both into saliva and water for 20 hrs. The highest concentration of BPA leaching from one out of 10 books was 43.4 ng/ml (for 2 hrs) in saliva, which was estimated to be approximately 0.052 mg/kg body weight/day for infants aged 6-10 months. As BPA has endocrine-disrupting effects and poses higher risks in infants than in adults, it is desired to reduce BPA use in the printing of infant books from the viewpoint of child health.

  18. A Qualitative Review on the Pharmacokinetics of Antibiotics in Saliva: Implications on Clinical Pharmacokinetic Monitoring in Humans.

    Science.gov (United States)

    Kiang, Tony K L; Ensom, Mary H H

    2016-03-01

    We conducted a systematic search to describe the current state of knowledge regarding the utility of saliva for clinical pharmacokinetic monitoring (CPM) of antibiotics. Although the majority of identified studies lacked sufficient pharmacokinetic data needed to assign an appropriate suitability classification, most aminoglycosides, fluoroquinolones, macrolides, penicillins/cephalosporins, and tetracyclines are likely not suitable for CPM in saliva. No clear pattern of correlation was observed between physiochemical properties that favor drug distribution into saliva and the likelihood of the antibiotic being classified as suitable for CPM in saliva (and vice versa). Insufficient data were available to determine if pathophysiological conditions affected salivary distribution of antibiotics. Additional confirmatory data are required for drugs (especially in patients) that are deemed likely suitable for CPM in saliva because only a few studies were available and many focused only on healthy subjects. All studies identified had relatively small sample sizes and exhibited large variability. Very few studies reported salivary collection parameters (e.g., salivary flow, pH) that could potentially have some impact on drug distribution into saliva. The available data are heavily weighted on healthy subjects, and insufficient data were available to determine if pathophysiology had effects on saliva drug distribution. Some studies also lacked assay sensitivity for detecting antibiotics in saliva. Overall, this review can be useful to clinicians who desire an overview on the suitability of saliva for conducting CPM of specific antibiotics, or for researchers who wish to fill the identified knowledge gaps to move the science of salivary CPM further.

  19. Salivary pH: A diagnostic biomarker

    Directory of Open Access Journals (Sweden)

    Sharmila Baliga

    2013-01-01

    Full Text Available Objectives: Saliva contains a variety of host defense factors. It influences calculus formation and periodontal disease. Different studies have been done to find exact correlation of salivary biomarkers with periodontal disease. With a multitude of biomarkers and complexities in their determination, the salivary pH may be tried to be used as a quick chairside test. The aim of this study was to analyze the pH of saliva and determine its relevance to the severity of periodontal disease. Study Design: The study population consisted of 300 patients. They were divided into three groups of 100 patients each: Group A had clinically healthy gingiva, Group B who had generalized chronic gingivitis and Group C who had generalized chronic periodontitis. The randomized unstimulated saliva from each patient was collected and pH was tested. Data was analyzed statistically using analysis of variance technique. Results: The salivary pH was more alkaline for patients with generalized chronic gingivitis as compared with the control group (P = 0.001 whereas patients with generalized chronic periodontitis had more acidic pH as compared with the control group (P = 0.001. Conclusion: These results indicate a significant change in the pH depending on the severity of the periodontal condition. The salivary pH shows significant changes and thus relevance to the severity of periodontal disease. Salivary pH may thus be used as a quick chairside diagnostic biomarker.

  20. Optimization of nonribosomal peptides production by a psychrotrophic fungus: Trichoderma velutinum ACR-P1.

    Science.gov (United States)

    Sharma, Richa; Singh, Varun P; Singh, Deepika; Yusuf, Farnaz; Kumar, Anil; Vishwakarma, Ram A; Chaubey, Asha

    2016-11-01

    Trichoderma is an anamorphic filamentous fungal genus with immense potential for production of small valuable secondary metabolites with indispensable biological activities. Microbial dynamics of a psychrotrophic strain Trichoderma velutinum ACR-P1, isolated from unexplored niches of the Shiwalik region, bestowed with rich biodiversity of microflora, was investigated for production of nonribosomal peptides (NRPs) by metabolite profiling by intact-cell mass spectrometry (ICMS) employing matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometer. Being the first report on NRPs production by T. velutinum, studies on optimization of growth conditions by Response Surface Methodology (RSM) for production of NRPs by ACR-P1 was carried out strategically. Multifold enhancement in the yield of NRPs belonging to subfamily SF4 with medium chain of amino acid residues having m/z 1437.9, 1453.9, and 1452.0 at pH 5.9 at 20 °C and of subfamily SF1 with long-chain amino acid residues having m/z 1770.2, 1784.2, 1800.1, 1802.1, and 1815.1 was achieved at pH 7.0 at 25 °C. Complexities of natural mixtures were thus considerably reduced under respective optimized culture conditions accelerating the production of novel microbial natural products by saving time and resources.

  1. Unstimulated salivary flow rate, pH and buffer capacity of saliva in healthy volunteers.

    Science.gov (United States)

    Fenoll-Palomares, C; Muñoz Montagud, J V; Sanchiz, V; Herreros, B; Hernández, V; Mínguez, M; Benages, A

    2004-11-01

    To assess the salivary flow rate, pH, and buffer capacity of healthy volunteers, and their relationships with age, gender, obesity, smoking, and alcohol consumption, and to establish the lower-end value of normal salivary flow (oligosialia). A prospective study was conducted in 159 healthy volunteers (age > 18 years, absence of medical conditions that could decrease salivary flow). Unstimulated whole saliva was collected during ten minutes, and salivary flow rate (ml/min), pH, and bicarbonate concentration (mmol/l) were measured using a Radiometer ABL 520. The 5 percentile of salivary flow rate and bicarbonate concentration was considered the lower limit of normality. Median salivary flow rate was 0.48 ml/min (range: 0.1-2 ml/min). Age younger than 44 years was associated with higher flow rates (OR 2.10). Compared with women, men presented a higher flow rate (OR 3.19) and buffer capacity (OR 2.81). Bicarbonate concentration correlated with salivary flow rate. The lower-end values of normal flow rate and bicarbonate concentration were 0.15 ml/min and 1.800 mmol/l, respectively. The presence of obesity, smoking, and alcohol consumption did not influence salivary parameters. In healthy volunteers, salivary flow rate depends on age and gender, and correlates with buffer capacity. Obesity, smoking, and alcohol use do not influence salivary secretion.

  2. Scalp hair and saliva as biomarkers in determination of mercury levels in Iranian women: Amalgam as a determinant of exposure

    Energy Technology Data Exchange (ETDEWEB)

    Fakour, H., E-mail: fakour.h@gmail.com [Department of Environment, Faculty of Natural Resources and Marine Sciences, Tarbiat Modares University, Noor, Mazandaran (Iran, Islamic Republic of); Esmaili-Sari, A. [Department of Environment, Faculty of Natural Resources and Marine Sciences, Tarbiat Modares University, Noor, Mazandaran (Iran, Islamic Republic of); Zayeri, F. [Department of Biostatistics, Faculty of Paramedical Sciences and Proteomics Research Center, Shahid Beheshti University of Medical Sciences, Tehran (Iran, Islamic Republic of)

    2010-05-15

    The aim of this study was to determine the relationship between mercury concentrations in saliva and hair in women with amalgam fillings and its relation with age and number of amalgam fillings. Eighty-two hair and saliva samples were collected randomly from Iranian women who have the same fish consumption pattern and free from occupational exposures. The mean {+-} SD age of these women was 29.37 {+-} 8.12 (ranged from 20 to 56). The determination of Hg level in hair samples was carried out by the LECO, AMA 254, Advanced Mercury Analyzer according to ASTM, standard No. D-6722. Mercury concentration in saliva samples was analyzed by PERKIN-ELMER 3030 Cold Vapor Atomic Absorption Spectrophotometer. The mean {+-} SD mercury level in the women was 1.28 {+-} 1.38 {mu}g/g in hair and 4.14 {+-} 4.08 {mu}g/l in saliva; and there were positive correlation among them. A significant correlation was also observed between Hg level of saliva (Spearman's {rho} = 0.93, P < 0.001) and hair (Spearman's {rho} = 0.92, P < 0.001) with number of amalgam fillings. According to the results, we can conclude that amalgam fillings may be an effective source for high Hg concentration in hair and releasing the mercury to the saliva samples.

  3. Effect of Saliva Contamination on Microleakage Beneath Bonded Brackets: A Comparison between Two Moisture-Tolerant Bonding Systems

    Directory of Open Access Journals (Sweden)

    Mohammad Hossein Toodehzaeim

    2016-03-01

    Full Text Available Objectives: This study aimed to evaluate the effects of saliva contamination on the metallic bracket microleakage bonded with two moisture-tolerant bonding systems.Materials and Methods:  Ninety freshly extracted premolar teeth were randomly divided into six groups of 15 with the following treatments: G1 (control: After acid etching, Assure primer and Assure adhesive were applied to non-contaminated enamel surfaces. G2 (contaminated after etching: The etched enamel surface was exposed to saliva, then Assure primer and Assure adhesive were applied. G3 (contaminated after priming: Saliva contamination was done after application of Assure primer. The exact same procedures were applied to groups G4 to G6 except that TIMP primer and Transbond Plus adhesive system were used.  To measure the microleakage score, the teeth were stained with 2% methylene blue for 24 hours, sectioned and examined under a stereomicroscope at ×16 magnification. Data analysis was performed using Fisher’s exact test.Results: In dry conditions, Assure and TMIP were not significantly different in terms of microleakage scores.  All contaminated groups exhibited higher microleakage score at the enamel/adhesive interface compared to the bracket/adhesive interface (P< 0.01. In wet conditions, Assure groups showed higher microleakage at the enamel-adhesive interface compared to the TMIP groups (P<0.05. At the bracket-adhesive interface, the microleakage scores were not significantly different in saliva contaminated groups compared to the controls. Conclusion: Saliva contamination caused greater microleakage at the enamel-adhesive interface compared to the adhesive-bracket interface.Keywords: Orthodontic Brackets; Adhesives; Saliva

  4. Suitability of saliva cortisol as a biomarker for hypothalamic-pituitary-adrenal axis activation assessment, effects of feeding actions, and immunostimulatory challenges in dairy cows

    DEFF Research Database (Denmark)

    Schwinn, A. -C.; Knight, C. H.; Bruckmaier, R. M.

    2016-01-01

    , current research focuses on noninvasive sampling of media other than blood, for example, saliva. The aim of this study was to assess the suitability of saliva cortisol as a biomarker under different physiological and immunological states in dairy cows. Our objectives were to 1) evaluate the relationship...... between HPA axis activation and saliva cortisol concentration, 2) investigate effects of some feeding action (as influenced by feed and water consumption) on saliva cortisol concentration, and 3) evaluate the time lag between plasma and saliva cortisol during induced inflammatory conditions...... administration. Saliva and blood samples were taken before, during, and after drinking, feeding, and ruminating. Only a low correlation between saliva and plasma cortisol concentrations (r = 0.03, P = 0.83) but no significant effects of the different feeding actions on saliva cortisol were observed. When...

  5. Hepatitis B virus DNA in saliva from children with chronic hepatitis B infection: implications for saliva as a potential mode of horizontal transmission

    DEFF Research Database (Denmark)

    Heiberg, Ida Louise; Hoegh, Mette; Ladelund, Steen

    2010-01-01

    To explore the mechanism of horizontal transmission of hepatitis B virus (HBV) among children, we investigated the quantitative relationship between HBV in saliva and blood from 46 children with chronic hepatitis B. We found high levels of HBV DNA in saliva of HBeAg (+) children, suggesting saliva...... as a vehicle for horizontal transmission of HBV among children....

  6. Saliva with reduced calcium and phosphorous concentrations: Effect on erosion dental lesions.

    Science.gov (United States)

    Denucci, Giovanna Corrêa; Mantilla, Taís Fonseca; Amaral, Flávia Lucisano Botelho; Basting, Roberta Tarkany; França, Fabiana Mantovani Gomes; Turssi, Cecilia Pedroso

    2018-02-08

    To investigate whether saliva formulations with reduced calcium (Ca) and inorganic phosphorous (Pi) concentration would affect dental erosion caused by hydrochloric acid (HCl). Enamel and root dentine bovine slabs were embedded, polished and measured for surface Knoop microhardness (SMH). After reference areas were created, specimens were exposed to HCl solution (0.01M; pH 2; 120s) and immersed in artificial salivas (6h) containing three different Ca/Pi concentrations (n=15), which simulate serum conditions of normo-, mild- or severe hypocalcaemia. The control group was immersed in Ca/Pi-free saliva. The study protocol was carried out 2x/day for 5 days. Surface loss of enamel and root dentine was assessed using an optical profilometer and SMH was remeasured for enamel. ANOVA (pdental substrate, salivary formulations resembling serum hypocalcaemia affected surface loss due to erosion and rehardening thereof. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  7. The role of sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P) in the trafficking of normal and malignant cells

    Science.gov (United States)

    Ratajczak, Mariusz Z.; Suszynska, Malwina; Borkowska, Sylwia; Ratajczak, Janina; Schneider, Gabriela

    2014-01-01

    Introduction A common feature of many types of cells is their responsiveness to chemotactic gradients of factors for which they express the corresponding receptors. The most studied chemoattractants so far are peptide-based growth factors and a family of cytokines endowed with strong chemotactic properties, called chemokines. However, additional evidence has accumulated that, in addition to these peptide-based chemoattractants, an important role in cell migration is played by bioactive lipids. Areas covered Solid evidence has accumulated that two bioactive phosphorylated sphingolipids that are derivatives of sphingolipid metabolism, namely sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P), are potent chemoattractants for a variety of cells. In this review, we will discuss the effect of these two phosphorylated sphingolipids on the trafficking of normal and malignant cells, and, in particular, we will focus on their role in trafficking of normal hematopoietic stem/progenitor cells. Unlike other mediators, S1P under steady state conditions maintain a steep gradient between interstitial fluid and peripheral blood and lymph across the endothelial barrier, which is important in the egress of cells from bone marrow. Both S1P and C1P may be upregulated in damaged tissues, which may result in reversal of this gradient. Expert opinion S1P and C1P are important regulators of the trafficking of normal and malignant cells, and modification of their biological effects will have important applications in optimizing stem cell mobilization and homing, tissue organ/regeneration, and preventing cancer metastasis. PMID:24188167

  8. Comparative analysis of biological activities of Der p I-derived peptides on Fc epsilon receptor-bearing cells from Dermatophagoides pteronyssinus-sensitive patients.

    Science.gov (United States)

    Jeannin, P; Pestel, J; Bossus, M; Lassalle, P; Tartar, A; Tonnel, A B

    1993-01-01

    The ability of four uncoupled synthetic peptides (p52-71, p117-133, p176-187, p188-199) derived from Der p I, a major allergen from the house dust mite Dermatophagoides pteronyssinus (Dpt) to stimulate Fc epsilon R+ cells from Dpt-sensitive patients was comparatively analysed. Each free peptide may specifically stimulate basophils (Fc epsilon RI+ cells) and platelets (Fc epsilon RII+ cells) from patients with significant levels of anti-Der p I IgE antibodies; p52-71 and p117-133 appear the best cell stimulation inducers. Both concentration-dependent biological activities of Der p I-peptide on Fc epsilon R+ cells are enhanced by coupling peptide to a carrier (as human serum albumin). Interestingly each Der p I-sensitive patient tested presents an individual pattern of response to peptide. Thus, from our results it appears that different Der p I sequences could be involved in the immune response to Der p I. PMID:7682161

  9. Comparative analysis of biological activities of Der p I-derived peptides on Fc epsilon receptor-bearing cells from Dermatophagoides pteronyssinus-sensitive patients.

    Science.gov (United States)

    Jeannin, P; Pestel, J; Bossus, M; Lassalle, P; Tartar, A; Tonnel, A B

    1993-04-01

    The ability of four uncoupled synthetic peptides (p52-71, p117-133, p176-187, p188-199) derived from Der p I, a major allergen from the house dust mite Dermatophagoides pteronyssinus (Dpt) to stimulate Fc epsilon R+ cells from Dpt-sensitive patients was comparatively analysed. Each free peptide may specifically stimulate basophils (Fc epsilon RI+ cells) and platelets (Fc epsilon RII+ cells) from patients with significant levels of anti-Der p I IgE antibodies; p52-71 and p117-133 appear the best cell stimulation inducers. Both concentration-dependent biological activities of Der p I-peptide on Fc epsilon R+ cells are enhanced by coupling peptide to a carrier (as human serum albumin). Interestingly each Der p I-sensitive patient tested presents an individual pattern of response to peptide. Thus, from our results it appears that different Der p I sequences could be involved in the immune response to Der p I.

  10. Effect of Synthetic Truncated Apolipoprotein C-I Peptide on Plasma Lipoprotein Cholesterol in Nonhuman Primates

    Directory of Open Access Journals (Sweden)

    Rampratap S. Kushwaha

    2004-01-01

    Full Text Available The present studies were conducted to determine whether a synthetic truncated apoC-I peptide that inhibits CETP activity in baboons would raise plasma HDL cholesterol levels in nonhuman primates with low HDL levels. We used 2 cynomolgus monkeys and 3 baboons fed a cholesterol- and fat-enriched diet. In cynomolgus monkeys, we injected synthetic truncated apoC-I inhibitor peptide at a dose of 20 mg/kg and, in baboons, at doses of 10, 15, and 20 mg/kg at weekly intervals. Blood samples were collected 3 times a week and VLDL + LDL and HDL cholesterol concentrations were measured. In cynomolgus monkeys, administration of the inhibitor peptide caused a rapid decrease in VLDL + LDL cholesterol concentrations (30%–60% and an increase in HDL cholesterol concentrations (10%–20%. VLDL + LDL cholesterol concentrations returned to baseline levels in approximately 15 days. In baboons, administration of the synthetic inhibitor peptide caused a decrease in VLDL + LDL cholesterol (20%–60% and an increase in HDL cholesterol (10%–20%. VLDL + LDL cholesterol returned to baseline levels by day 21, whereas HDL cholesterol concentrations remained elevated for up to 26 days. ApoA-I concentrations increased, whereas apoE and triglyceride concentrations decreased. Subcutaneous and intravenous administrations of the inhibitor peptide had similar effects on LDL and HDL cholesterol concentrations. There was no change in body weight, food consumption, or plasma IgG levels of any baboon during the study. These studies suggest that the truncated apoC-I peptide can be used to raise HDL in humans.

  11. The levels of bioelements in the saliva and hair of young athletes

    Directory of Open Access Journals (Sweden)

    N. A. Troegubova

    2016-01-01

    Full Text Available The salivary and hair levels of bioelements, such as calcium, potassium, magnesium, iron, zinc, chromium, and selenium, were investigated in 130 children and adolescents, including 35 swimmers, 47 hockey players, 20 fencers, and 28 schoolchildren who did not additionally go in for sports. Atomic emission spectrometry and inductively coupled plasma mass spectrometry were applied. The aim of our study was to investigate the characteristics of macro- and microelemental composition of the saliva and hair of young athletes. The investigation established that that all young athletes showed the high levels of potassium and selenium in their hair, those of selenium and chromium in the saliva, and a significantly low concentration of potassium in the saliva (p<0,05, reflecting the intensity of mineral exchange during physical exercises. Individual characteristics were found in the content of chemical elements in the test biological substrates in relation to the kind of sports: the salivary levels of calcium and magnesium were reduced and the hair level of iron was increased in fencers (p<0,05; and the field-hockey players had higher zinc values (p<0,05. Comparison of the hair and salivary concentrations of bioelements established that there was a statistically significant relationship for potassium (R=-0,72; p<0,001 and selenium (R=0,44; p=0,004. Non-invasive screening studies of salivary and hair minerals are recommended for the early diagnosis of an imbalance in macro- and microelements in young athletes. There is evidence that young athletes should be identified as a group of patients at risk for an imbalance of macro- and microelements.

  12. A hierarchical coarse-grained (all-atom to all residue) approach to peptides (P1, P2) binding with a graphene sheet

    Science.gov (United States)

    Pandey, Ras; Kuang, Zhifeng; Farmer, Barry; Kim, Sang; Naik, Rajesh

    2012-02-01

    Recently, Kim et al. [1] have found that peptides P1: HSSYWYAFNNKT and P2: EPLQLKM bind selectively to graphene surfaces and edges respectively which are critical in modulating both the mechanical as well as electronic transport properties of graphene. Such distinctions in binding sites (edge versus surface) observed in electron micrographs were verified by computer simulation by an all-atomic model that captures the pi-pi bonding. We propose a hierarchical approach that involves input from the all-atom Molecular Dynamics (MD) study (with atomistic detail) into a coarse-grained Monte Carlo simulation to extend this study further to a larger scale. The binding energy of a free amino acid with the graphene sheet from all-atom simulation is used in the interaction parameter for the coarse-grained approach. Peptide chain executes its stochastic motion with the Metropolis algorithm. We investigate a number of local and global physical quantities and find that peptide P1 is likely to bind more strongly to graphene sheet than P2 and that it is anchored by three residues ^4Y^5W^6Y. [1] S.N. Kim et al J. Am. Chem. Soc. 133, 14480 (2011).

  13. Bone induction by biomimetic PLGA copolymer loaded with a novel synthetic RADA16-P24 peptide in vivo

    International Nuclear Information System (INIS)

    Pan, Haitao; Hao, Shaofei; Zheng, Qixin; Li, Jingfeng; Zheng, Jin; Hu, Zhilei; Yang, Shuhua; Guo, Xiaodong; Yang, Qin

    2013-01-01

    Bone morphogenetic protein-2 (BMP-2) is a key bone morphogenetic protein, and poly(lactic-co-glycolic acid) (PLGA) has been widely used as scaffold for clinical use to carry treatment protein. In the previous studies, we have synthesized BMP-2-related peptide (P24) and found its capacity of inducing bone regeneration. In this research, we have synthesized a new amphiphilic peptide Ac-RADA RADA RADA RADA S[PO4]KIPKASSVPTELSAISTLYLDDD-CONH2 (RADA16-P24) with an assembly peptide RADA16-Ion the P24 item of BMP2 to form divalent ion-induced gelatin. Two methods of physisorption and chemical cross-linking were used to bind RADA16-P24 onto the surface of the copolymer PLGA to synthesize RADA16-P24–PLGA, and its capacity of attaching bone marrow stromal cells (BMSCs) was evaluated in vitro and inducing ectopic bone formation was examined in vivo. In vitro our results demonstrated that RADA16-P24–PLGA copolymer prepared by physisorbing or prepared by chemical cross-linking had a peptide binding rate of (2.0180 ± 0.5296)% or (10.0820 ± 0.8405)% respectively (P < 0.05). In addition the BMSCs proliferated vigorously in the RADA16-P24–PLGA biomaterials. Significantly the percentage of BMSCs attached to RADA16-P24–PLGA composite prepared by chemical cross-linking and physisorbing were (71.4 ± 7.5) % or (46.7 ± 5.8) % (P < 0.05). The in vivo study showed that RADA16-P24–PLGA chemical cross-linking could better induce ectopic bone formation compared with RADA16-P24–PLGA physisorbing and PLGA. It is concluded that the PLGA copolymer is a good RADA16-P24 carrier. This novel RADA16-P24–PLGA composite has strong osteogenic capability. - Highlights: • We have synthesized a new RADA16-P24 amphiphilic peptide. • It is an assembly peptide RADA16-Ion the P24 to form divalent ion-induced gelatin. • RADA16-P24/PLGA could better induce etopia osteogenesis compared with PLGA. • RADA16-P24–PLGA has strong osteogenic capability

  14. Bone induction by biomimetic PLGA copolymer loaded with a novel synthetic RADA16-P24 peptide in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Pan, Haitao; Hao, Shaofei [Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022 (China); Zheng, Qixin, E-mail: zheng-qx@163.com [Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022 (China); Li, Jingfeng [Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022 (China); Department of Orthopedics, Zhongnan Hospital of Wuhan University, Wuhan 430071 (China); Zheng, Jin; Hu, Zhilei; Yang, Shuhua; Guo, Xiaodong [Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022 (China); Yang, Qin [Advanced Biomaterials and Tissue Engineering Center, Huazhong University of Science and Technology, Wuhan 430074 (China)

    2013-08-01

    Bone morphogenetic protein-2 (BMP-2) is a key bone morphogenetic protein, and poly(lactic-co-glycolic acid) (PLGA) has been widely used as scaffold for clinical use to carry treatment protein. In the previous studies, we have synthesized BMP-2-related peptide (P24) and found its capacity of inducing bone regeneration. In this research, we have synthesized a new amphiphilic peptide Ac-RADA RADA RADA RADA S[PO4]KIPKASSVPTELSAISTLYLDDD-CONH2 (RADA16-P24) with an assembly peptide RADA16-Ion the P24 item of BMP2 to form divalent ion-induced gelatin. Two methods of physisorption and chemical cross-linking were used to bind RADA16-P24 onto the surface of the copolymer PLGA to synthesize RADA16-P24–PLGA, and its capacity of attaching bone marrow stromal cells (BMSCs) was evaluated in vitro and inducing ectopic bone formation was examined in vivo. In vitro our results demonstrated that RADA16-P24–PLGA copolymer prepared by physisorbing or prepared by chemical cross-linking had a peptide binding rate of (2.0180 ± 0.5296)% or (10.0820 ± 0.8405)% respectively (P < 0.05). In addition the BMSCs proliferated vigorously in the RADA16-P24–PLGA biomaterials. Significantly the percentage of BMSCs attached to RADA16-P24–PLGA composite prepared by chemical cross-linking and physisorbing were (71.4 ± 7.5) % or (46.7 ± 5.8) % (P < 0.05). The in vivo study showed that RADA16-P24–PLGA chemical cross-linking could better induce ectopic bone formation compared with RADA16-P24–PLGA physisorbing and PLGA. It is concluded that the PLGA copolymer is a good RADA16-P24 carrier. This novel RADA16-P24–PLGA composite has strong osteogenic capability. - Highlights: • We have synthesized a new RADA16-P24 amphiphilic peptide. • It is an assembly peptide RADA16-Ion the P24 to form divalent ion-induced gelatin. • RADA16-P24/PLGA could better induce etopia osteogenesis compared with PLGA. • RADA16-P24–PLGA has strong osteogenic capability.

  15. Modulation of taste responsiveness by the satiation hormone peptide YY

    Science.gov (United States)

    La Sala, Michael S.; Hurtado, Maria D.; Brown, Alicia R.; Bohórquez, Diego V.; Liddle, Rodger A.; Herzog, Herbert; Zolotukhin, Sergei; Dotson, Cedrick D.

    2013-01-01

    It has been hypothesized that the peripheral taste system may be modulated in the context of an animal's metabolic state. One purported mechanism for this phenomenon is that circulating gastrointestinal peptides modulate the functioning of the peripheral gustatory system. Recent evidence suggests endocrine signaling in the oral cavity can influence food intake (FI) and satiety. We hypothesized that these hormones may be affecting FI by influencing taste perception. We used immunohistochemistry along with genetic knockout models and the specific reconstitution of peptide YY (PYY) in saliva using gene therapy protocols to identify a role for PYY signaling in taste. We show that PYY is expressed in subsets of taste cells in murine taste buds. We also show, using brief-access testing with PYY knockouts, that PYY signaling modulates responsiveness to bitter-tasting stimuli, as well as to lipid emulsions. We show that salivary PYY augmentation, via viral vector therapy, rescues behavioral responsiveness to a lipid emulsion but not to bitter stimuli and that this response is likely mediated via activation of Y2 receptors localized apically in taste cells. Our findings suggest distinct functions for PYY produced locally in taste cells vs. that circulating systemically.—La Sala, M. S., Hurtado, M. D., Brown, A. R., Bohórquez, D. V., Liddle, R. A., Herzog, H., Zolotukhin, S., Dotson, C. D. Modulation of taste responsiveness by the satiation hormone peptide YY. PMID:24043261

  16. Facilitated saliva secretion and reduced oral inflammation by a novel artificial saliva system in the treatment of salivary hypofunction

    Directory of Open Access Journals (Sweden)

    Kang M

    2017-01-01

    Full Text Available Minkyung Kang,1 Hyounggeun Park,1 Joon-Ho Jun,1 Miwon Son,1 Myung Joo Kang2 1Pharmaceutical Product Research Laboratories, Dong-A ST Research Institute, Gyeonggi, 2Division of Pharmaceutical Sciences, College of Pharmacy, Dankook University, Cheonan, Chungnam, Korea Abstract: Saliva substitutes and/or lubricants are commonly employed to lessen dry mouth symptoms by stimulating and/or substituting for the secretion of saliva. In this study, a novel artificial saliva containing inorganic salts, including sodium chloride and potassium chloride, and bactericidal agents, including potassium thiocyanate and lactoperoxidase, was formulated in the form of a solution (DM-sol or gel (DM-gel. Those in vivo therapeutic efficacies were assessed in terms of saliva secretion and anti-inflammatory activity in rats and mice, respectively. Salivary secretion was promoted by mucosal application of DM-formulations in normal rats. In particular, DM-gel resulted in 2.5- and 1.9-fold greater salivary flow rates compared to normal saline and DM-sol, respectively. In an in vivo efficacy evaluation in diabetic mice with salivary hypofunction, repeated application of DM-formulations alleviated histopathological changes in the buccal mucosa in terms of atrophy and thinning of the epithelium, compared to vehicle, after 4 weeks. Moreover, the DM-sol and DM-gel were comparably effective for relieving periodontal gingivitis, reducing infiltration of inflammatory cells, and normalizing the neutrophil level in the gingival gingiva, after 4 weeks. Therefore, the novel artificial saliva is expected to facilitate salivary secretion and restore physiological conditions in the mouth of patients with salivary hypofunction. Keywords: saliva substitute, carbopol gel, hypothiocyanite–hydrogen peroxide mixture, antimicrobial activity, diabetic rats

  17. Disposable collection kit for rapid and reliable collection of saliva.

    Science.gov (United States)

    Yamaguchi, Masaki; Tezuka, Yuki; Takeda, Kazunori; Shetty, Vivek

    2015-01-01

    To describe and evaluate disposable saliva collection kit for rapid, reliable, and reproducible collection of saliva samples. The saliva collection kit comprised of a saliva absorbent swab and an extractor unit was used to retrieve whole saliva samples from 10 subjects. The accuracy and precision of the extracted volumes (3, 10, and 30 μl) were compared to similar volumes drawn from control samples obtained by passive drool. Additionally, the impact of kit collection method on subsequent immunoassay results was verified by assessing salivary cortisol levels in the samples and comparing them to controls. The recovered volumes for the whole saliva samples were 3.85 ± 0.28, 10.79 ± 0.95, and 31.18 ± 1.72 μl, respectively (CV = 8.76%) and 2.91 ± 0.19, 9.75 ± 0.43, and 29.64 ± 0.91 μl, respectively, (CV = 6.36%) for the controls. There was a close correspondence between the salivary cortisol levels from the saliva samples obtained by the collection kit and the controls (R(2)  > 0.96). The disposable saliva collection kit allows accurate and repeatable collection of fixed amounts of whole saliva and does not interfere with subsequent measurements of salivary cortisol. The simple collection process, lack of elaborate specimen recovery steps, and the short turnaround time (<3 min) should render the kit attractive to test subjects and researchers alike. © 2015 Wiley Periodicals, Inc.

  18. Rationally designed chimeric peptide of met-enkephalin and FMRFa-[D-Ala2,p-Cl-Phe4]YFa induce multiple opioid receptors mediated antinociception and up-regulate their expression.

    Science.gov (United States)

    Vats, Ishwar Dutt; Chaudhary, Snehlata; Sharma, Ahuti; Nath, Mahendra; Pasha, Santosh

    2010-07-25

    The physiological role of NPFF/FMRFa family of peptides appears to be complex and exact mechanism of action of these peptides is not yet completely understood. In same line of scrutiny, another analog of YGGFMKKKFMRFamide (YFa), a chimeric peptide of met-enkephalin and FMRFamide, was rationally designed and synthesized which contain D-alanine and p-Cl-phenylalanine residues at 2nd and 4th positions, respectively i.e., Y-(D-Ala)-G-(p-Cl-Phe)-MKKKFMRFamide ([D-Ala(2), p-Cl-Phe(4)]YFa) in order to achieve improved bioavailability and blood brain barrier penetration. Therefore, present study investigates the possible antinociceptive effect of [D-Ala(2), p-Cl-Phe(4)]YFa on intra-peritoneal (i.p.) administration using tail-flick test in rats followed by its opioid receptor(s) specificity using mu, delta and kappa receptor antagonists. Further, its antinociceptive effect was examined during 6 days of chronic i.p. treatment and assessed effect of this treatment on differential expression of opioid receptors. [D-Ala(2), p-Cl-Phe(4)]YFa in comparison to parent peptide YFa, induce significantly higher dose dependent antinociception in rats which was mediated by all three opioid receptors (mu, delta and kappa). Importantly, it induced comparable antinociception in rats throughout the chronic i.p. treatment and significantly up-regulated the overall expression (mRNA and protein) of mu, delta and kappa opioid receptors. Therefore, pharmacological and molecular behavior of [D-Ala(2), p-Cl-Phe(4)]YFa demonstrate that incorporation of D-alanine and p-Cl-phenylalanine residues at appropriate positions in chimeric peptide leads to altered opioid receptor selectivity and enhanced antinociceptive potency, relative to parent peptide. (c) 2010 Elsevier B.V. All rights reserved.

  19. C- and N-truncated antimicrobial peptides from LFampin 265 - 284: Biophysical versus microbiology results

    Directory of Open Access Journals (Sweden)

    Regina Adão

    2011-01-01

    Full Text Available Lactoferrin is a glycoprotein with two globular lobes, each having two domains. Since the discovery of its antimicrobial properties, efforts have been made to find peptides derived from this protein showing antimicrobial properties. Most peptides initially studied were derived from Lactoferricin B, obtained from the protein by digestion with pepsin. More recently, a new family of antimicrobial peptides (AMPs derived from Lactoferrin was discovered by Bolcher et al, and named Lactoferrampin (LFampin. The original sequence of LFampin contained residues 268 - 284 from the N1 domain of Lactoferrin. From this peptide, the Bolscher′s group synthesized a collection of peptides obtained by extension and / or truncation at the C or N-terminal sides, in order to unravel the main structural features responsible for antimicrobial action. Here, we present results for three of these peptides, namely LFampin 265 - 284, LFampin 265 - 280, and LFampin 270 - 284. The peptides were tested against bacteria (E. coli and S. sanguinis, fungi (C. albicans, and model membranes of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC, 1,2-dimyristoyl-sn-glycero-3-[phospho-rac-(1-glycerol] (DMPG, and their mixtures at a ratio of 3 : 1 (DMPC : DMPG (3 : 1. The ability to adopt a helical conformation was followed by a circular dichroism (CD, and the perturbation of the gel to the liquid-crystalline phase transition of the membrane was characterized by differential scanning calorimetry (DSC. Distinct behavior was observed in the three peptides, both from the microbiology and model membrane studies, with the biophysical results showing excellent correlation with the microbiology activity studies. LFampin 265 - 284 was the most active peptide toward the tested microorganisms, and in the biophysical studies it showed the highest ability to form an a-helix and the strongest interaction with model membranes, followed by LFampin 265 - 280. LFampin 270 - 284 was inactive, showing

  20. Fluoride in dental biofilm and saliva

    DEFF Research Database (Denmark)

    Larsen, Line Staun

    Dette ph.d.-projekt bidrager med ny viden om fordelingen af fluorid i dental biofilm og saliva. For at udforske koncentrationen af fluorid i naturlig (in vivo) biofilmvæske, biofilmsediment og i saliva, blev der udført to meget forskellige kliniske studier. Resultaterne fra tværsnitsstudiet (Studie...... I), hos en stor gruppe mennesker (n=42) der konsulterede en tandklinik for behandling, bekræfter tidligere viden, at der findes en naturlig biologisk variation i fluoridkoncentrationerne i biofilm fra forskellige intra-orale regioner samt mellem biofilmvæske, biofilmsediment og saliva...... fluoridkoncentrationer i underkæbefronten, intermediære koncentrationer i alle tre overkæberegioner og de laveste koncentrationer i underkæbemolarregionerne. Begge studier viser at biofilmsedimentet indeholder størstedelen af fluorid i biofilm. Set i et bredere perspektiv viser fundene at der er et omvendt forhold...

  1. The diagnostic value of pepsin detection in saliva for gastro-esophageal reflux disease: a preliminary study from China.

    Science.gov (United States)

    Du, Xing; Wang, Feng; Hu, Zhiwei; Wu, Jimin; Wang, Zhonggao; Yan, Chao; Zhang, Chao; Tang, Juan

    2017-10-17

    None of current diagnostic methods has been proven to be a reliable tool for gastro-esophageal reflux disease (GERD). Pepsin in saliva has been proposed as a promising diagnostic biomarker for gastro-esophageal reflux. We aimed to determine the diagnostic value of salivary pepsin detection for GERD. Two hundred and fifty patients with symptoms suggestive of GERD and 35 asymptomatic healthy volunteers provided saliva on morning waking, after lunch and dinner for pepsin determination using the Peptest lateral flow device. All patients underwent 24-h multichannel intraluminal impedance pH (24-h MII-pH) monitoring and upper gastrointestinal endoscopy. Based on 24-h MII-pH and endoscopy study, patients were defined as GERD (abnormal MII-pH results and/or reflux esophagitis) and non-GERD otherwise. Patients with GERD had a higher prevalence of pepsin in saliva and higher pepsin concentration than patients with non-GERD and healthy controls (P < 0.001 for all). The pepsin test had a sensitivity of 73% and a specificity of 88.3% for diagnosing GERD using the optimal cut-off value of 76 ng/mL. Postprandial saliva samples collected when the symptoms occurred had a more powerful ability to identify GERD. Salivary pepsin test had moderate diagnostic value for GERD. It may be a promising tool to replace the use of currently invasive tools with advantages of non-invasive, easy to perform and cost effective. ChiCTR-DDD-16009506 (date of registration: October 20, 2016).

  2. Structural and pharmacological characteristics of chimeric peptides derived from peptide E and beta-endorphin reveal the crucial role of the C-terminal YGGFL and YKKGE motifs in their analgesic properties.

    Science.gov (United States)

    Condamine, Eric; Courchay, Karine; Rego, Jean-Claude Do; Leprince, Jérôme; Mayer, Catherine; Davoust, Daniel; Costentin, Jean; Vaudry, Hubert

    2010-05-01

    Peptide E (a 25-amino acid peptide derived from proenkephalin A) and beta-endorphin (a 31-amino acid peptide derived from proopiomelanocortin) bind with high affinity to opioid receptors and share structural similarities but induce analgesic effects of very different intensity. Indeed, whereas they possess the same N-terminus Met-enkephalin message sequence linked to a helix by a flexible spacer and a C-terminal part in random coil conformation, in contrast with peptide E, beta-endorphin produces a profound analgesia. To determine the key structural elements explaining this very divergent opioid activity, we have compared the structural and pharmacological characteristics of several chimeric peptides derived from peptide E and beta-endorphin. Structures were obtained under the same experimental conditions using circular dichroism, computational estimation of helical content and/or nuclear magnetic resonance spectroscopy (NMR) and NMR-restrained molecular modeling. The hot-plate and writhing tests were used in mice to evaluate the antinociceptive effects of the peptides. Our results indicate that neither the length nor the physicochemical profile of the spacer plays a fundamental role in analgesia. On the other hand, while the functional importance of the helix cannot be excluded, the last 5 residues in the C-terminal part seem to be crucial for the expression or absence of the analgesic activity of these peptides. These data raise the question of the true function of peptides E in opioidergic systems. Copyright (c) 2010 Elsevier Inc. All rights reserved.

  3. Pelepasan ion nikel dan kromium kawat Australia dan stainless steel dalam saliva buatan (The release of nickel and chromium ions from Australian wire and stainless steel in artificial saliva

    Directory of Open Access Journals (Sweden)

    Nolista Indah Rasyid

    2014-09-01

    Full Text Available Background: Fixed orthodontic treatment needs several types of wire to produce biomechanical force to move teeth. The use orthodontic wire within the mouth interacts with saliva, causing the release of nickel and chromium ions. Purpose: The study was aimed to examine the effect of immersion time in artificial saliva between special type of Australian wire and stainless steel on the release of nickel and chromium ions. Methods: Thirty special type Australian wires and 30 stainless steel wires were used in this study, each of which weighed 0.12 grams. The wires were immersed for 1, 7, 28, 35, 42, and 49 days in artificial saliva with a normal pH. The release of ions in saliva was examined using Atomic Absorption spectrophotometry. Results: The result indicated that the release of nickel ions on special type of Australian wire was larger than that on stainless steel wire (p<0.005, there were differences in the release of the amount of nickel ions on special type of Australia in different immersion time, and there was a correlation between the types of wire and immersion time. Nickel ions released from the special type of Australian wire detected on the 7th day of immersion and reached its peak on the 35th day, while from stainless steel wire were detected on the 49th day of immersion. The released of chromium ions from the special type of Australian wire and stainless steel wire were not detected until the 49th day of immersion. Conclusion: The release of nickel ions were highest on the 35th day of immersion in special type of Australian wire and they were detected on the 49th day in stainless steel wire. The release of chromium ions were not detected until 49th day of immersion in special type of Australian and stainless steel wire.Latar belakang: Perawatan ortodonti cekat memerlukan beberapa macam kawat untuk menghasilkan kekuatan biomekanika yang sesuai dalam menggerakkan gigi. Pemakaian kawat ortodonti di dalam mulut dapat bereaksi dengan

  4. Radioimmunological method for determination of cortisol in saliva

    International Nuclear Information System (INIS)

    Maleeva, A.; Mileva, Zh.; Kekhajova, M.

    1989-01-01

    A method was developed for determination of cortisol in saliva after being previously extracted with dichlormethane. Cortisol concentration in saliva of 19 subjects was determined by this method. The saliva cortisol levels were compared with those of blood plasma. No statistically significant difference was found. The method finds acceptance primarily when frequent measurements of cortisol level are neccessary as a screening technique and when strongly abnormally high levels should be differentiated from the normal ones: in this latter case determination of plasma cortisol is mandatory. 5 tabs., 8 refs

  5. Effects of saliva collection using cotton swab on cortisol enzyme immunoassay.

    Science.gov (United States)

    Kozaki, Tomoaki; Hashiguchi, Nobuko; Kaji, Yumi; Yasukouchi, Akira; Tochihara, Yutaka

    2009-12-01

    Cotton swabs are among the most commonly used devices for collecting saliva, but various studies have reported that their use impacts the results of salivary cortisol assays. These studies, however, estimated this impact by comparing the average of the concentration and/or scatter plots. In the present study, we estimated the impact of cotton swabs on the results of salivary cortisol enzyme immunoassay (EIA) by Bland-Altman plot. Eight healthy males (aged 20-23 years) provided four saliva samples on different days to yield a total of 32 samples. Saliva samples were collected directly in plastic tubes using plastic straws and then pipetted onto cotton swabs (cotton saliva collection) and into clear sterile tubes (passive saliva collection). There was a lower correlation between cotton and passive saliva collection. Individually, four subjects showed a negative correlation between passive and cotton saliva collection. A Bland-Altman plot indicated that cotton swabs causes a proportional bias on the EIA assay result. Our findings indicate a considerable effect of using cotton swabs for saliva collection, and subject-specific variability in the impact. A Bland-Altman plot further suggests possible reasons for this effect.

  6. A Study of the Insulin and the C-Peptide Responses to Oral Glucose Load in Nondiabetic and Diabetic Subjects

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Myung Chul; Choi, Sung Jae; Kim, Eung Jin; Koh, Chang Soon; Min, Hun Ki [Seoul National University College of Medicine, Seoul (Korea, Republic of)

    1977-03-15

    The present study was undertaken to evaluate the significance of the insulin and the C-peptide response to oral glucose loads in normal and diabetic subjects and to establish the effects of the obesity. In this study, the authors have measured plasma insulin and C-peptide by means of radioimmunoassay in 10 nonobese normal, 5 obese normal, 13 nonobese moderate diabetic patients, 9 obese moderate diabetic patients and 9 severe diabetic patients. The results obtained were as follows; 1) In 10 nonobese normal subjects, the plasma insulin level at fasting state and at 30, 60, 90, and 120 min after oral glucose loads were 15.7+-3.4, 48.3+-9.8, 40.4+-6.7, 37.4+-6.5 and 26.0+-4.2 uU/ml (Mean+-S.E.) and C-peptide were 1.9+-0.3, 3.9+-0.6, 6.3+-0.6, 5.7+-0.5 and 4.0+-0.5 ng/ml. The change of C-peptide was found to go almost parallel with that of insulin and the insulin value reaches to the highest level at 30 min whereas C-peptide reaches to its peak at 60 min.. 2) The plasma insulin level in 5 obese normal subjects were 38.9+-12.3, 59.5+-12.3, 59.2+-17.1, 56.1+-20.0 and 48.4+-17.2 uU/ml and the C-peptide were 5.5+-0.4, 6.8+-0.5, 7.9+-0.8, 7.9+-0.8 and 7.8+-2.0 ng/ml. The insulin response appeared to be greater than nonobese normal subjects. 3) In 13 nonobese moderate diabetic patients, the plasma insulin levels were 27.1+-4.9, 44.1+-6.0, 37.3+-6.6, 35.5+-8.1 and 34.7+-10.7 uU/ml and the C-peptide levels were 2.7+-0.4, 4.9+-0.7, 6.5+-0.5, 7.0+-0.3 and 6.7+-1.0 ng/ml. There was little significance compared to nonobese normal groups but delayed pattern is noted. 4) In 9 obese moderated diabetic patients, the plasma insulin levels were 22.1+-7.9, 80.0+-19.3, 108.0+-27.0, 62.0+-17.6 and 55.5+-10.l uU/ml and the C-peptide levels were 5.2+-0.4, 8.0+-1.0, 10.4+-1.6, 10.4+-1.7 and 10.1+-1.0 ng/ml and its response was also greater than that of nonobese moderate diabetic patients. 5) The plasma insulin concentrations in 9 severe diabetic subjects were 8.0+-3.8, 12.1+-3.5, 16.8+-4.6, 19

  7. Use of Fructosyl Peptide Oxidase for HbA1c Assay

    Science.gov (United States)

    Yonehara, Satoshi; Inamura, Norio; Fukuda, Miho; Sugiyama, Koji

    2015-01-01

    ARKRAY, Inc developed the world’s first automatic glycohemoglobin analyzer based on HPLC (1981). After that, ARKRAY developed enzymatic HbA1c assay “CinQ HbA1c” with the spread and diversification of HbA1c measurement (2007). CinQ HbA1c is the kit of Clinical Chemistry Analyzer, which uses fructosyl peptide oxidase (FPOX) for a measurement reaction. This report mainly indicates the developmental background, measurement principle, and future of the enzymatic method HbA1c reagent. PMID:25633966

  8. Scalp hair and saliva as biomarkers in determination of mercury levels in Iranian women: Amalgam as a determinant of exposure

    International Nuclear Information System (INIS)

    Fakour, H.; Esmaili-Sari, A.; Zayeri, F.

    2010-01-01

    The aim of this study was to determine the relationship between mercury concentrations in saliva and hair in women with amalgam fillings and its relation with age and number of amalgam fillings. Eighty-two hair and saliva samples were collected randomly from Iranian women who have the same fish consumption pattern and free from occupational exposures. The mean ± SD age of these women was 29.37 ± 8.12 (ranged from 20 to 56). The determination of Hg level in hair samples was carried out by the LECO, AMA 254, Advanced Mercury Analyzer according to ASTM, standard No. D-6722. Mercury concentration in saliva samples was analyzed by PERKIN-ELMER 3030 Cold Vapor Atomic Absorption Spectrophotometer. The mean ± SD mercury level in the women was 1.28 ± 1.38 μg/g in hair and 4.14 ± 4.08 μg/l in saliva; and there were positive correlation among them. A significant correlation was also observed between Hg level of saliva (Spearman's ρ = 0.93, P < 0.001) and hair (Spearman's ρ = 0.92, P < 0.001) with number of amalgam fillings. According to the results, we can conclude that amalgam fillings may be an effective source for high Hg concentration in hair and releasing the mercury to the saliva samples.

  9. The corrosion behaviour of austenitic and duplex stainless steels in artificial saliva with the addition of fluoride

    International Nuclear Information System (INIS)

    Kocijan, Aleksandra; Merl, Darja Kek; Jenko, Monika

    2011-01-01

    Research highlights: → The corrosion behaviour of AISI 316L and 2205 DSS in orthodontics. → The increased passive range for DSS 2205 compared to AISI 316L in artificial saliva. → Higher R p values of DSS compared to AISI 316L in artificial saliva. → The main constituent of the passive layers on DSS at the OCP in saliva was Cr-oxide. → DSS 2205 is suitable for orthodontic applications in artificial saliva. - Abstract: The evolution of the passive films on 2205 duplex stainless steel (2205 DSS) and AISI 316L stainless steel in artificial saliva, and with the addition of fluoride, was studied using electrochemical impedance spectroscopy (EIS) and potentiodynamic measurements. The extent of the passive range increased for the 2205 DSS compared to the AISI 316L in both solutions. The formation of the passive film was studied by EIS at the open-circuit potential (OCP). The passive layers were studied at the OCP by X-ray photoelectron spectroscopy (XPS). The passive films on both materials predominantly contained Cr-oxides, whereas the Fe species were markedly depleted.

  10. The corrosion behaviour of austenitic and duplex stainless steels in artificial saliva with the addition of fluoride

    Energy Technology Data Exchange (ETDEWEB)

    Kocijan, Aleksandra, E-mail: Aleksandra.Kocijan@imt.s [Institute of Metals and Technology, Lepi pot 11, 1000 Ljubljana (Slovenia); Merl, Darja Kek [Jozef Stefan Institute, Jamova 39, 1000 Ljubljana (Slovenia); Jenko, Monika [Institute of Metals and Technology, Lepi pot 11, 1000 Ljubljana (Slovenia)

    2011-02-15

    Research highlights: The corrosion behaviour of AISI 316L and 2205 DSS in orthodontics. The increased passive range for DSS 2205 compared to AISI 316L in artificial saliva. Higher R{sub p} values of DSS compared to AISI 316L in artificial saliva. The main constituent of the passive layers on DSS at the OCP in saliva was Cr-oxide. DSS 2205 is suitable for orthodontic applications in artificial saliva. - Abstract: The evolution of the passive films on 2205 duplex stainless steel (2205 DSS) and AISI 316L stainless steel in artificial saliva, and with the addition of fluoride, was studied using electrochemical impedance spectroscopy (EIS) and potentiodynamic measurements. The extent of the passive range increased for the 2205 DSS compared to the AISI 316L in both solutions. The formation of the passive film was studied by EIS at the open-circuit potential (OCP). The passive layers were studied at the OCP by X-ray photoelectron spectroscopy (XPS). The passive films on both materials predominantly contained Cr-oxides, whereas the Fe species were markedly depleted.

  11. Total anti-oxidant capacity of saliva in chronic periodontitis patients before and after periodontal treatment.

    Science.gov (United States)

    Shirzaiy, M; Ansari, S M; Dehghan, J H; Ghaeni, S H

    2014-01-01

    Periodontal disease is among the most common inflammatory conditions which is associated with many different factors. One of the contributing factors to the pathogenesis of this condition may compromise the defensive mechanism of antioxidants. The present study evaluates the antioxidant capacity of saliva in periodontal patients before and after periodontal treatment. In this cross sectional study, 31 patients systemically healthy non smokers with chronic periodontitis were recruited. The antioxidant capacity of saliva was measured before the initial phase of periodontal therapy and after completion of the treatment. Data were analyzed using SPSS 19 software. Paired T-Test, Independent sample T-test and ANOVA tests were used as appropriated. The mean and standard deviation antioxidant capacity of the saliva after the treatment.(0.962± 0.287µM)was significantly higher than before the treatment (0.655 ± 0.281 µM ,pperiodontal treatment was higher among men than among women; however, the difference was not significant (P=0.07). The mean difference of salivary antioxidant capacity was not significantly differed among different ages (P=0.772). The antioxidant capacity of saliva was higher after periodontal therapy among patients with periodontal disease, however the change was not varied across the ages and gender. Therefore, the alterations in the defensive mechanism of antioxidants could be the key factors contribute to the pathogenesis of periodontal diseases.

  12. Noninvasive glucose monitoring using saliva nano-biosensor

    Directory of Open Access Journals (Sweden)

    Wenjun Zhang

    2015-06-01

    Full Text Available Millions of people worldwide live with diabetes and several millions die from it each year. A noninvasive, painless method of glucose testing would highly improve compliance and glucose control while reducing complications and overall disease management costs. To provide accurate, low cost, and continuous glucose monitoring, we have developed a unique, disposable saliva nano-biosensor. More than eight clinical trials on real-time noninvasive salivary glucose monitoring were carried out on two healthy individuals (a 2–3 h-period for each trial, including both regular food and standard glucose beverage intake with more than 35 saliva samples obtained. Excellent clinical accuracy was revealed as compared to the UV Spectrophotometer. By measuring subjects’ salivary glucose and blood glucose in parallel, we found the two generated profiles share the same fluctuation trend but the correlation between them is individual dependent. There is a time lag between the peak glucose values from blood and from saliva. However, the correlation between the two glucose values at fasting is constant for each person enabling noninvasive diagnosis of diabetes through saliva instead of blood. Furthermore, a good correlation of glucose levels in saliva and in blood before and 2 h after glucose intake was observed. Glucose monitoring before and 2 h after meals is usually prescribed by doctors for diabetic patients. Thus, this disposable biosensor will be an alternative for real-time salivary glucose tracking at any time.

  13. The role of insulin C-peptide in the coevolution analyses of the insulin signaling pathway: a hint for its functions.

    Directory of Open Access Journals (Sweden)

    Shuai Wang

    Full Text Available As the linker between the A chain and B chain of proinsulin, C-peptide displays high variability in length and amino acid composition, and has been considered as an inert byproduct of insulin synthesis and processing for many years. Recent studies have suggested that C-peptide can act as a bioactive hormone, exerting various biological effects on the pathophysiology and treatment of diabetes. In this study, we analyzed the coevolution of insulin molecules among vertebrates, aiming at exploring the evolutionary characteristics of insulin molecule, especially the C-peptide. We also calculated the correlations of evolutionary rates between the insulin and the insulin receptor (IR sequences as well as the domain-domain pairs of the ligand and receptor by the mirrortree method. The results revealed distinctive features of C-peptide in insulin intramolecular coevolution and correlated residue substitutions, which partly supported the idea that C-peptide can act as a bioactive hormone, with significant sequence features, as well as a linker assisting the formation of mature insulin during synthesis. Interestingly, the evolution of C-peptide exerted the highest correlation with that of the insulin receptor and its ligand binding domain (LBD, implying a potential relationship with the insulin signaling pathway.

  14. Shear bond strength of orthodontic brackets and disinclusion buttons: effect of water and saliva contamination.

    Science.gov (United States)

    Sfondrini, Maria Francesca; Fraticelli, Danilo; Gandini, Paola; Scribante, Andrea

    2013-01-01

    The aim of this study was to assess the effect of water and saliva contamination on the shear bond strength and failure site of orthodontic brackets and lingual buttons. 120 bovine permanent mandibular incisors were randomly divided into 6 groups of 20 specimens each. Both orthodontic brackets and disinclusion buttons were tested under three different enamel surface conditions: (a) dry, (b) water contamination, and (c) saliva contamination. Brackets and buttons were bonded to the teeth and subsequently tested using a Instron universal testing machine. Shear bond strength values and adhesive failure rate were recorded. Statistical analysis was performed using ANOVA and Tukey tests (strength values) and Chi squared test (ARI Scores). Noncontaminated enamel surfaces showed the highest bond strengths for both brackets and buttons. Under water and saliva contamination orthodontic brackets groups showed significantly lower shear strengths than disinclusion buttons groups. Significant differences in debond locations were found among the groups under the various enamel surface conditions. Water and saliva contamination of enamel during the bonding procedure lowers bond strength values, more with orthodontic brackets than with disinclusion buttons.

  15. Rehardening of caries-like lesions in root surfaces by saliva substitutes.

    Science.gov (United States)

    Turssi, Cecilia P; Lima, Renata Q V; Faraoni-Romano, Juliana J; Serra, Mônica C

    2006-12-01

    This study sought to evaluate whether root dentine caries-like lesions could be remineralised by saliva substitutes. Root dentine slabs (3 x 3 x 2 mm) were cut from bovine incisors, ground flat, polished and pre-tested for Knoop microhardness (KHN) at five locations spaced 500 microm apart and 500 microm from the left edge of each sectioned piece. After 60 out of the 100 slabs had been selected based upon their KHN values, specimens were coated with wax except for their outer surface. Specimens were then cycled through a highly cariogenic challenge model to induce caries-like lesions, whose formation was confirmed by KHN measurements located 500 microm from the right edge of the specimen. According to a randomised complete block design, the experimental units (n = 15) were exposed to 1.5 ml of saliva substitutes, based on either mucin (MC) or carboxymethylcellulose (CM), to natural human saliva (HS) or to 100% relative humidity (RH) over 20 days. Remineralisation was verified by KHN measurements located 1000 microm apart from the right edge of the specimen. Analysis of variance indicated a significant (p < 0.0001) difference among the KHN values attained by the carious root dentine after exposure to the remineralising agents. Tukey's test ascertained that remineralisation was greatest with MC, intermediate with CM and least with HS, but rehardening did not reach the pre-caries lesion formation values. Saliva substitutes may provide partial remineralisation to preformed caries-like lesions in root dentine.

  16. Influência da contaminação com saliva na microinfiltração de restaurações de resina composta = The effect of saliva contamination on microleakage of composite restorative

    Directory of Open Access Journals (Sweden)

    Hasan, Nihad Hasan Musa

    2005-01-01

    Full Text Available O objetivo deste estudo foi avaliar a influência da contaminação com saliva na microinfiltração marginal de restaurações adesivas. Utilizaram-se 52 incisivos bovinos, sendo confeccionadas cavidades Classe V (vestibular/lingual, com margens em dentina e esmalte. As cavidades foram condicionadas com ácido fosfórico a 35% e divididas em quatro grupos. G1 – sem contaminação, os demais grupos foram contaminados com saliva fresca por 15s, sendo submetidos a diferentes tratamentos: G2 – secas com papel absorvente; G3 – lavadas com água; G4 – recondicionadas por 10s. O sistema adesivo (Single Bond/3M ESPE foi empregado segundo recomendações do fabricante e as cavidades restauradas com compósito (Filtek Z-250/3M ESPE. Após 24h as restaurações foram polidas e termocicladas (500 ciclos/5 e 55ºC/30s. Os dentes foram isolados, exceto as restaurações e a 1mm destas, imersos em fucsina básica a 1% (24h e lavados em água (24h. Foram seccionados e avaliados em lupa estereoscópica, utilizando-se escores predeterminados, por dois examinadores calibrados. Os valores obtidos foram submetidos à análise estatística (Kruskal-Wallis. A infiltração em dentina foi significantemente maior que aquela observada em esmalte (p < 0,05. Nas margens em dentina observou-se redução do selamento no G2 (p < 0,05. G3 e G4 restabeleceram o selamento a níveis similares ao controle (G1. Nas margens em esmalte G1 apresentou selamento similar ao G3 e ambos exibiram melhor selamento que G2 e G4 (p < 0,05, que foram similares. Conclusões: dentro das limitações do estudo, verificou-se que a contaminação com saliva após o condicionamento ácido interfere no selamento marginal de restaurações adesivas e os tratamentos propostos apresentaram resultados diversos

  17. Squalestatin alters the intracellular trafficking of a neurotoxic prion peptide

    Directory of Open Access Journals (Sweden)

    Williams Alun

    2007-11-01

    Full Text Available Abstract Background Neurotoxic peptides derived from the protease-resistant core of the prion protein are used to model the pathogenesis of prion diseases. The current study characterised the ingestion, internalization and intracellular trafficking of a neurotoxic peptide containing amino acids 105–132 of the murine prion protein (MoPrP105-132 in neuroblastoma cells and primary cortical neurons. Results Fluorescence microscopy and cell fractionation techniques showed that MoPrP105-132 co-localised with lipid raft markers (cholera toxin and caveolin-1 and trafficked intracellularly within lipid rafts. This trafficking followed a non-classical endosomal pathway delivering peptide to the Golgi and ER, avoiding classical endosomal trafficking via early endosomes to lysosomes. Fluorescence resonance energy transfer analysis demonstrated close interactions of MoPrP105-132 with cytoplasmic phospholipase A2 (cPLA2 and cyclo-oxygenase-1 (COX-1, enzymes implicated in the neurotoxicity of prions. Treatment with squalestatin reduced neuronal cholesterol levels and caused the redistribution of MoPrP105-132 out of lipid rafts. In squalestatin-treated cells, MoPrP105-132 was rerouted away from the Golgi/ER into degradative lysosomes. Squalestatin treatment also reduced the association between MoPrP105-132 and cPLA2/COX-1. Conclusion As the observed shift in peptide trafficking was accompanied by increased cell survival these studies suggest that the neurotoxicity of this PrP peptide is dependent on trafficking to specific organelles where it activates specific signal transduction pathways.

  18. Saliva as a future potential predictor for various periodontal diseases

    Directory of Open Access Journals (Sweden)

    Zahreni Hamzah

    2011-06-01

    Full Text Available Background: There are many diagnostic biomarkers have been found in saliva. Saliva contains a wide variety of proteins, including bacteria and products, enzymes, inflammatory mediators and host response modifiers, products of tissue breakdown. Purpose: The purpose of the study was studied current development of diagnostic biomarkers in saliva that will lead to the development of simple and accurate diagnostic tools for periodental disease. Reviews: Specifically, the salivary biomarkers divided for three aspects of periodontitis i.e. inflammation, collagen degradation and bone turnover, correlated with clinical features of periodontal disease. The diagnostic biomarkers is in saliva, such as enzyme, immunoglobulin, cytokines, bacteria and bacterial products, hormones. For the past two decades, oral health researchers have been developing salivary diagnostic tools to monitor oral diseases. Conclusion: The indicators of acute periodontitis can detect with ß-glucuronidase and AST, IL-1β, and MMP-8, whereas indicators for chronic periodontitis can detect with ALP. The indicators for collagen degradation and bone turnover suggest ICTP, fibronectin fragments, and osteonectin. The indicators of severity of periodontitis especially can be predict by B. forsythus.Latar belakang: Banyak biomarker telah ditemukan dalam saliva. Saliva terdiri dari berbagai protein unik meliputi bakteri dan produk bakteri, enzim, mediator inflamasi dan modifikasi respon host (immunoglobulin, sitokin, produk kerusakan jaringan (telopeptida kolagen, osteokalsin, proteoglikan, fragmen fibronectin. Tujuan: Mengkaji biomarker dalam saliva untuk pengembangan metode diagnostik sederhana dan akurat untuk penyakit periodontal. Tinjauan Pustaka: Secara khusus, biomarker saliva pada periodontitis dibagi dalam tiga aspek yaitu inflamasi, dan degradasi kolagen serta pergantian tulang. Biomarker diagnostik dalam saliva, meliputi enzim, imunoglobulin, sitokin, bakteri dan produk

  19. What peptides these deltorphins be.

    Science.gov (United States)

    Lazarus, L H; Bryant, S D; Cooper, P S; Salvadori, S

    1999-02-01

    The deltorphins are a class of highly selective delta-opioid heptapeptides from the skin of the Amazonian frogs Phyllomedusa sauvagei and P. bicolor. The first of these fascinating peptides came to light in 1987 by cloning of the cDNA of from frog skins, while the other members of this family were identified either by cDNA or isolation of the peptides. The distinctive feature of deltorphins is the presence of a naturally occurring D-enantiomer at the second position in their common N-terminal sequence, Tyr-D-Xaa-Phe, comparable to dermorphin, which is the prototype of a group of mu-selective opioids from the same source. The D-amino acid and the anionic residues, either Glu or Asp, as well as their unique amino acid compositions are responsible for the remarkable biostability, high delta-receptor affinity, bioactivity and peptide conformation. This review summarizes a decade of research from many laboratories that defined which residues and substituents in the deltorphins interact with the delta-receptor and characterized pharmacological and physiological activities in vitro and in vivo. It begins with a historical description of the topic and presents general schema for the synthesis of peptide analogues of deltorphins A, B and C as a means to document the methods employed in producing a myriad of analogues. Structure activity studies of the peptides and their pharmacological activities in vitro are detailed in abundantly tabulated data. A brief compendium of the current level of knowledge of the delta-receptor assists the reader to appreciate the rationale for the design of these analogues. Discussion of the conformation of these peptides addresses how structure leads to further hypotheses regarding ligand receptor interaction. The review ends with a broad discussion of the potential applications of these peptides in clinical and therapeutic settings.

  20. Analysis of changes of serum leptin, C-peptide levels and peripheral fat tissue leptin receptor expression in obesity

    International Nuclear Information System (INIS)

    Du Tongxin; Sun Junjiang; Wang Shukui; Fu Lei

    2002-01-01

    Objective: To explore the mechanism of obesity and obesity accompanied type two diabetes mellitus by investigating changes of serum leptin, C-peptide (C-P) levels and leptin receptor expression in peripheral adipose tissues. Methods: Peripheral leptin receptor density was measured via radio-ligand binding method, serum leptin and C - P levels were measured via radioimmunoassay in 91 cases (38 in obesity group, 23 in over weight, and 30 in normal controls). Results: With the increase of body mass index (BMI), the peripheral leptin receptor density of the over weight and obese cases decreased and was mash less than that of normal cases (both p<0.01, respectively). There was no statistical differences for Kd value among the three groups, suggesting no associated change between the binding ability of leptin receptor to its ligand. There was a negative correlation between BMI and leptin receptor density (r = -0.70, p < 0.01). The serum leptin and C-P levels in weight excess and obese subjects with type two DM were both increased, but significantly higher in obese group than those in weight excess group (p < 0.01). The increase of C-P was much marked than that of leptin. Serum C-P level was positively correlated with BMI. Conclusion: Changes of serum leptin, C-P levels and peripheral leptin receptor expression in cases with simple obesity and obesity accompanied with type two DM were related closely with BMI. Type 2 DM in obese subjects was related with leptin resistance and insulin resistance

  1. Metabolic risk profiles in diabetes stratified according to age at onset, islet autoimmunity and fasting C-peptide

    DEFF Research Database (Denmark)

    Wod, Mette; Yderstræde, Knud B; Halekoh, Ulrich

    2017-01-01

    OBJECTIVE: Islet autoimmunity, age at onset and time to insulin treatment are often used to define subgroups of diabetes. However, the latter criterion is not clinical useful. Here, we examined whether an unbiased stratification of diabetes according to age at onset, fasting C-peptide and GAD......, fasting C-peptide above or below 300 pmol/l (CPEPhigh or CPEPlow), and presence or absence of GADab (GADpos or GADneg). HbA1c, BMI, blood pressure (BP), lipid profile, alanine aminotransferase (ALT) and creatinine were evaluated. RESULTS: GADab were present in 13% of the cohort. Age at onset...... as GADposCPEPhigh; n=327) and patients with type 2 diabetes (GADnegCPEPhigh; n=3,544). Patients with LADA defined an intermediate group with higher HbA1c but otherwise lower cardiometabolic risk than patients with type 2 diabetes. CONCLUSIONS: Our results demonstrate that fasting C-peptide and GADab status...

  2. Understanding of xerostomia and strategies for the development of artificial saliva.

    Science.gov (United States)

    Kho, Hong-Seop

    2014-01-01

    Xerostomia is becoming a major issue in dental and medical clinics with an increase of aged population. Medication is the most common etiology of xerostomia, while the most severe xerostomia generally occurs in patients with a history of head and neck radiotherapy. Xerostomic patients usually suffer from diminished quality of life due to various symptoms and complications. Decreased salivary output is a definite objective sign, but oral mucosal wetness is a more reliable factor for the evaluation of xerostomia. At present there are no effective therapeutic methods for the treatment of xerostomia. Sialogogues may have problematic side effects and their therapeutic effects last only brief duration. Artificial saliva typically does not produce satisfactory results in therapeutic efficacy. Therefore, further research and development of better therapeutic modalities are necessary. The basic concept for the development of ideal and functional artificial saliva is the mimicry of natural human saliva. We need proper candidate molecules and antimicrobial supplements to simulate the rheological and biological properties of human saliva. We also need better understanding of the interactions between the ingredients of artificial saliva themselves and between the ingredients and components of human saliva both in solution and on surface phases. In addition, we need accepted measures to evaluate the efficacy of artificial saliva. In conclusion, for the development of ideal artificial saliva, research based on the understanding of pathophysiology of xerostomia and knowledge about rheological and biological functions of human saliva are necessary.

  3. Periodontitis diagnostics using resonance Raman spectroscopy on saliva

    Science.gov (United States)

    Gonchukov, S.; Sukhinina, A.; Bakhmutov, D.; Biryukova, T.; Tsvetkov, M.; Bagratashvily, V.

    2013-07-01

    In view of its wealth of molecular information, Raman spectroscopy has been the subject of active biomedical research. The aim of this work is Raman spectroscopy (RS) application for the determination of molecular biomarkers in saliva with the objective of early periodontitis detection. As was shown in our previous study, carotenoids contained in saliva can be molecular fingerprint information for the periodontitis level. It is shown here that the carotenoid RS lines at wavenumbers of 1156 and 1524 cm-1 can be easily detected and serve as reliable biomarkers of periodontitis using resonance Raman spectroscopy of dry saliva.

  4. Periodontitis diagnostics using resonance Raman spectroscopy on saliva

    International Nuclear Information System (INIS)

    Gonchukov, S; Sukhinina, A; Bakhmutov, D; Biryukova, T; Tsvetkov, M; Bagratashvily, V

    2013-01-01

    In view of its wealth of molecular information, Raman spectroscopy has been the subject of active biomedical research. The aim of this work is Raman spectroscopy (RS) application for the determination of molecular biomarkers in saliva with the objective of early periodontitis detection. As was shown in our previous study, carotenoids contained in saliva can be molecular fingerprint information for the periodontitis level. It is shown here that the carotenoid RS lines at wavenumbers of 1156 and 1524 cm −1 can be easily detected and serve as reliable biomarkers of periodontitis using resonance Raman spectroscopy of dry saliva. (letter)

  5. Unstimulated salivary flow rate, pH and buffer capacity of saliva in healthy volunteers Débito basal, pH y capacidad tampón de la secreción salivar en sujetos sanos

    OpenAIRE

    C. Fenoll-Palomares; J. V. Muñoz-Montagud; V. Sanchiz; B. Herreros; V. Hernández; M. Mínguez; A. Benages

    2004-01-01

    Objectives: to assess the salivary flow rate, pH, and buffer capacity of healthy volunteers, and their relationships with age, gender, obesity, smoking, and alcohol consumption, and to establish the lower-end value of normal salivary flow (oligosialia). Methods: a prospective study was conducted in 159 healthy volunteers (age > 18 years, absence of medical conditions that could decrease salivary flow). Unstimulated whole saliva was collected during ten minutes, and salivary flow rate (ml/min)...

  6. The Effect of Canal Contamination with Saliva on Apical Sealing

    Directory of Open Access Journals (Sweden)

    S Sabaghi

    2014-08-01

    Full Text Available Introduction: Root canal obturation aims at sealing the root canal system to prevent re-contamination of canal and periapical space. Presence of moisture in canal before obturation may posit a negative effect on the quality of canal sealing. Therefore, this study was conducted to investigate the effect of canal contamination with saliva on apical microleakage. Methods: In this laboratory study, 58 human uni-root teeth were cleaned and shaped for obturation with gutta percha and sealer AH26. In the case group, specimens were contaminated with human saliva immediately before obturation, whereas the teeth in the control group were kept dry. All canals were filled by lateral condensation technique. Moreover, the teeth were placed in methylene blue dye for 3 days. Dye penetration was measured using a stereomicrosope. As a matter of fact, the study data were analyzed via utilizing t-test. Results: A significant difference was found between the two groups in regard with the apical leakage(P<0.001. The microleakage mean of dye in the dry group was 3/48mm, whereas it was 6/36mm in the saliva contaminated group. Conclusion: The study findings revealed that complete drying of canal can improve apical sealing.

  7. C P Gopalakrishnan

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education. C P Gopalakrishnan. Articles written in Resonance – Journal of Science Education. Volume 9 Issue 6 June 2004 pp 74-85 Classroom. The Real Effects of Pseudo Forces · P Chaitanya Das G Srinivasa Murthy C P Gopalakrishnan P C Deshmukh · More Details ...

  8. Avaliação do pH da saliva e da saburra lingual antes e após a utilização de soluções enxaguantes orais e sua relação com parâmetros de halitose

    OpenAIRE

    Elen de Souza Tolentino

    2009-01-01

    A relação entre halitose, pH da saliva e da saburra lingual frente ao uso de enxaguantes orais ainda não é totalmente conhecido. O objetivo deste estudo foi avaliar o pH da saliva e da saburra lingual em pacientes com saúde oral íntegra e halitose fisiológica através de um pHmetro analógico e um digital e de fitas indicadoras de pH, antes e imediatamente após utilização de diferentes enxaguantes orais e 30 minutos após o bochecho; avaliar o pH dos diferentes anti-sépticos; avaliar se há difer...

  9. Structural Analysis of Der p 1–Antibody Complexes and Comparison with Complexes of Proteins or Peptides with Monoclonal Antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Osinski, Tomasz; Pomés, Anna; Majorek, Karolina A.; Glesner, Jill; Offermann, Lesa R.; Vailes, Lisa D.; Chapman, Martin D.; Minor, Wladek; Chruszcz, Maksymilian (INDOOR); (UV); (SC)

    2015-05-29

    Der p 1 is a major allergen from the house dust mite, Dermatophagoides pteronyssinus, that belongs to the papain-like cysteine protease family. To investigate the antigenic determinants of Der p 1, we determined two crystal structures of Der p 1 in complex with the Fab fragments of mAbs 5H8 or 10B9. Epitopes for these two Der p 1–specific Abs are located in different, nonoverlapping parts of the Der p 1 molecule. Nevertheless, surface area and identity of the amino acid residues involved in hydrogen bonds between allergen and Ab are similar. The epitope for mAb 10B9 only showed a partial overlap with the previously reported epitope for mAb 4C1, a cross-reactive mAb that binds Der p 1 and its homolog Der f 1 from Dermatophagoides farinae. Upon binding to Der p 1, the Fab fragment of mAb 10B9 was found to form a very rare α helix in its third CDR of the H chain. To provide an overview of the surface properties of the interfaces formed by the complexes of Der p 1–10B9 and Der p 1–5H8, along with the complexes of 4C1 with Der p 1 and Der f 1, a broad analysis of the surfaces and hydrogen bonds of all complexes of Fab–protein or Fab–peptide was performed. This work provides detailed insight into the cross-reactive and specific allergen–Ab interactions in group 1 mite allergens. The surface data of Fab–protein and Fab–peptide interfaces can be used in the design of conformational epitopes with reduced Ab binding for immunotherapy.

  10. Inorganic component of saliva during fasting and after fast break

    OpenAIRE

    Samad, Rasmidar

    2016-01-01

    Oral health is closely related to salivary components. Saliva consists of water, inorganic and organic materials. Fasting changes one???s meal and drinking time that in turn can affect the environment in oral cavity, including inorganic componenet of saliva. The purpose of this study is to determine the inorganic component of saliva during fasting and after fast break.

  11. Prevalence of Pseudomonas aeruginosa and Acinetobacter spp. in subgingival biofilm and saliva of subjects with chronic periodontal infection

    Directory of Open Access Journals (Sweden)

    Renata Souto

    2014-06-01

    Full Text Available P. aeruginosa and Acinetobacter spp. are important pathogens associated with late nosocomial pneumonia in hospitalized and institutionalized individuals. The oral cavity may be a major source of these respiratory pathogens, particularly in the presence of poor oral hygiene and periodontal infection. This study investigated the prevalence of P. aeruginosa and Acinetobacter spp. in subgingival biofilm and saliva of subjects with periodontal disease or health. Samples were obtained from 55 periodontally healthy (PH and 169 chronic periodontitis (CP patients. DNA was obtained from the samples and detection of P. aeruginosa and Acinetobacter spp. was carried out by multiplex and nested PCR. P. aeruginosa and Acinetobacter spp. were detected in 40% and 45% of all samples, respectively. No significant differences in the distribution of these microorganisms between men and women, subgingival biofilm and saliva samples, patients 35 years of age, and smokers and non-smokers were observed regardless periodontal status (p > 0.05. In contrast, the frequencies of P. aeruginosa and Acinetobacter spp. in saliva and biofilm samples were significantly greater in CP than PH patients (p < 0.01. Smokers presenting P. aeruginosa and high frequencies of supragingival plaque were more likely to present CP than PH. P. aeruginosa and Acinetobacter spp. are frequently detected in the oral microbiota of CP. Poor oral hygiene, smoking and the presence of P. aeruginosa are strongly associated with periodontitis.

  12. A Study of the Insulin and the C-Peptide Responses to Oral Glucose Load in Nondiabetic and Diabetic Subjects

    International Nuclear Information System (INIS)

    Lee, Myung Chul; Choi, Sung Jae; Kim, Eung Jin; Koh, Chang Soon; Min, Hun Ki

    1977-01-01

    The present study was undertaken to evaluate the significance of the insulin and the C-peptide response to oral glucose loads in normal and diabetic subjects and to establish the effects of the obesity. In this study, the authors have measured plasma insulin and C-peptide by means of radioimmunoassay in 10 nonobese normal, 5 obese normal, 13 nonobese moderate diabetic patients, 9 obese moderate diabetic patients and 9 severe diabetic patients. The results obtained were as follows; 1) In 10 nonobese normal subjects, the plasma insulin level at fasting state and at 30, 60, 90, and 120 min after oral glucose loads were 15.7±3.4, 48.3±9.8, 40.4±6.7, 37.4±6.5 and 26.0±4.2 uU/ml (Mean±S.E.) and C-peptide were 1.9±0.3, 3.9±0.6, 6.3±0.6, 5.7±0.5 and 4.0±0.5 ng/ml. The change of C-peptide was found to go almost parallel with that of insulin and the insulin value reaches to the highest level at 30 min whereas C-peptide reaches to its peak at 60 min.. 2) The plasma insulin level in 5 obese normal subjects were 38.9±12.3, 59.5±12.3, 59.2±17.1, 56.1±20.0 and 48.4±17.2 uU/ml and the C-peptide were 5.5±0.4, 6.8±0.5, 7.9±0.8, 7.9±0.8 and 7.8±2.0 ng/ml. The insulin response appeared to be greater than nonobese normal subjects. 3) In 13 nonobese moderate diabetic patients, the plasma insulin levels were 27.1±4.9, 44.1±6.0, 37.3±6.6, 35.5±8.1 and 34.7±10.7 uU/ml and the C-peptide levels were 2.7±0.4, 4.9±0.7, 6.5±0.5, 7.0±0.3 and 6.7±1.0 ng/ml. There was little significance compared to nonobese normal groups but delayed pattern is noted. 4) In 9 obese moderated diabetic patients, the plasma insulin levels were 22.1±7.9, 80.0±19.3, 108.0±27.0, 62.0±17.6 and 55.5±10.l uU/ml and the C-peptide levels were 5.2±0.4, 8.0±1.0, 10.4±1.6, 10.4±1.7 and 10.1±1.0 ng/ml and its response was also greater than that of nonobese moderate diabetic patients. 5) The plasma insulin concentrations in 9 severe diabetic subjects were 8.0±3.8, 12.1±3.5, 16.8±4.6, 19

  13. Complex formation in mixtures of lysozyme-stabilized emulsions and human saliva

    NARCIS (Netherlands)

    Silletti, E.; Vingerhoeds, M.H.; Norde, W.; Aken, van G.A.

    2007-01-01

    In this paper, we studied the interaction between human unstimulated saliva and lysozyme-stabilized oil-in-water emulsions (10 wt/wt% oil phase, 10 mM NaCl, pH 6.7), to reveal the driving force for flocculation of these emulsions. Confocal scanning laser microscopy (CSLM) showed formation of

  14. Highly efficient synthetic method onpyroacm resin using the boc SPPS protocol for C-terminal cysteine peptide synthesis

    Energy Technology Data Exchange (ETDEWEB)

    Juvekar, Vinayak; Kim, Kang Tae; Gong, Young Dae [Innovative Drug Library Research Center, Dept. of Chemistry, College of Science, Dongguk University, Seoul (Korea, Republic of)

    2017-01-15

    A very effective process on Pyroacm resin was developed for solid-phase peptide synthesis (SPPS) of C-terminal cysteine and cysteine ester peptides. The process uses cysteine side chain anchoring to the Pyroacm resin and the Boc protocol for SPPS. The Pyroacm resin showed remarkable stability under standard trifluoromethanesulfonic acid (TFMSA) cleavage condition. TFMSA cleavage of protecting groups generates a peptide-linked resin, which can be subjected to peptide modification reactions. Finally, the peptide can be cleaved from the resin using methoxycarbonylsulfenyl chloride. The utility of this protocol was demonstrated by its applications to the synthesis of model peptides, key intermediates in the preparation of natural products riparin 1.2 and a-factor.

  15. Microbial community profiling of human saliva using shotgun metagenomic sequencing.

    Directory of Open Access Journals (Sweden)

    Nur A Hasan

    Full Text Available Human saliva is clinically informative of both oral and general health. Since next generation shotgun sequencing (NGS is now widely used to identify and quantify bacteria, we investigated the bacterial flora of saliva microbiomes of two healthy volunteers and five datasets from the Human Microbiome Project, along with a control dataset containing short NGS reads from bacterial species representative of the bacterial flora of human saliva. GENIUS, a system designed to identify and quantify bacterial species using unassembled short NGS reads was used to identify the bacterial species comprising the microbiomes of the saliva samples and datasets. Results, achieved within minutes and at greater than 90% accuracy, showed more than 175 bacterial species comprised the bacterial flora of human saliva, including bacteria known to be commensal human flora but also Haemophilus influenzae, Neisseria meningitidis, Streptococcus pneumoniae, and Gamma proteobacteria. Basic Local Alignment Search Tool (BLASTn analysis in parallel, reported ca. five times more species than those actually comprising the in silico sample. Both GENIUS and BLAST analyses of saliva samples identified major genera comprising the bacterial flora of saliva, but GENIUS provided a more precise description of species composition, identifying to strain in most cases and delivered results at least 10,000 times faster. Therefore, GENIUS offers a facile and accurate system for identification and quantification of bacterial species and/or strains in metagenomic samples.

  16. Saliva Polymerase-Chain-Reaction Assay for Cytomegalovirus Screening in Newborns

    Science.gov (United States)

    Boppana, Suresh B.; Ross, Shannon A.; Shimamura, Masako; Palmer, April L.; Ahmed, Amina; Michaels, Marian G.; Sánchez, Pablo J.; Bernstein, David I.; Tolan, Robert W.; Novak, Zdenek; Chowdhury, Nazma; Britt, William J.; Fowler, Karen B.

    2011-01-01

    BACKGROUND Congenital cytomegalovirus (CMV) infection is an important cause of hearing loss, and most infants at risk for CMV-associated hearing loss are not identified early in life because of failure to test for the infection. The standard assay for newborn CMV screening is rapid culture performed on saliva specimens obtained at birth, but this assay cannot be automated. Two alternatives — real-time polymerase-chain-reaction (PCR)–based testing of a liquid-saliva or dried-saliva specimen obtained at birth — have been developed. METHODS In our prospective, multicenter screening study of newborns, we compared real-time PCR assays of liquid-saliva and dried-saliva specimens with rapid culture of saliva specimens obtained at birth. RESULTS A total of 177 of 34,989 infants (0.5%; 95% confidence interval [CI], 0.4 to 0.6) were positive for CMV, according to at least one of the three methods. Of 17,662 newborns screened with the use of the liquid-saliva PCR assay, 17,569 were negative for CMV, and the remaining 85 infants (0.5%; 95% CI, 0.4 to 0.6) had positive results on both culture and PCR assay. The sensitivity and specificity of the liquid-saliva PCR assay were 100% (95% CI, 95.8 to 100) and 99.9% (95% CI, 99.9 to 100), respectively, and the positive and negative predictive values were 91.4% (95% CI, 83.8 to 96.2) and 100% (95% CI, 99.9 to 100), respectively. Of 17,327 newborns screened by means of the dried-saliva PCR assay, 74 were positive for CMV, whereas 76 (0.4%; 95% CI, 0.3 to 0.5) were found to be CMV-positive on rapid culture. Sensitivity and specificity of the dried-saliva PCR assay were 97.4% (95% CI, 90.8 to 99.7) and 99.9% (95% CI, 99.9 to 100), respectively. The positive and negative predictive values were 90.2% (95% CI, 81.7 to 95.7) and 99.9% (95% CI, 99.9 to 100), respectively. CONCLUSIONS Real-time PCR assays of both liquid- and dried-saliva specimens showed high sensitivity and specificity for detecting CMV infection and should be

  17. Aberrant proteins featured in the saliva of habitual betel quid chewers: an indication of early oral premalignancy?

    Science.gov (United States)

    Jessie, Kala; Jayapalan, Jaime Jacqueline; Rahim, Zubaidah Haji Abdul; Hashim, Onn Haji

    2014-12-01

    Prolonged chewing of betel quid is known to cause oral diseases, including cancer. The present study was performed to screen for aberrant proteins in the saliva of habitual betel quid chewers compared to nonchewers. Saliva of female subjects (n = 10) who had been chewing betel quid for more than 20 years and nonbetel quid chewers (n = 10) of the same gender and range of age was analyzed by gel-based proteomics. Increased structural microheterogeneity of saliva haptoglobin beta chains indicated by shifts of focused spots similar to that earlier reported in patients with oral squamous cell carcinoma, and their relatively higher abundance compared to nonbetel quid chewers, were detected in saliva protein profiles of all chewers. In addition, the majority of the betel quid chewers also showed significant higher abundance of hemopexin, alpha-1B glycoprotein, alpha1-antitrypsin, complement C3, and transthyretin. These proteins had previously been associated with several different cancers. Our data demonstrated different forms of protein aberration in the saliva of betel quid chewers, which may be indicative of early oral precancerous conditions. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Biochemical Characterisation of Phage Pseudomurein Endoisopeptidases PeiW and PeiP Using Synthetic Peptides.

    Science.gov (United States)

    Schofield, Linley R; Beattie, Amy K; Tootill, Catherine M; Dey, Debjit; Ronimus, Ron S

    2015-01-01

    Pseudomurein endoisopeptidases cause lysis of the cell walls of methanogens by cleaving the isopeptide bond Ala-ε-Lys in the peptide chain of pseudomurein. PeiW and PeiP are two thermostable pseudomurein endoisopeptidases encoded by phage ΨM100 of Methanothermobacter wolfei and phages ΨM1 and ΨM2 of Methanothermobacter marburgensis, respectively. A continuous assay using synthetic peptide substrates was developed and used in the biochemical characterisation of recombinant PeiW and PeiP. The advantages of these synthetic peptide substrates over natural substrates are sensitivity, high purity, and characterisation and the fact that they are more easily obtained than natural substrates. In the presence of a reducing agent, purified PeiW and PeiP each showed similar activity under aerobic and anaerobic conditions. Both enzymes required a divalent metal for activity and showed greater thermostability in the presence of Ca(2+). PeiW and PeiP involve a cysteine residue in catalysis and have a monomeric native conformation. The kinetic parameters, K(M) and k(cat), were determined, and the ε-isopeptide bond between alanine and lysine was confirmed as the bond lysed by these enzymes in pseudomurein. The new assay may have wider applications for the general study of peptidases and the identification of specific methanogens susceptible to lysis by specific pseudomurein endoisopeptidases.

  19. Biochemical Characterisation of Phage Pseudomurein Endoisopeptidases PeiW and PeiP Using Synthetic Peptides

    Directory of Open Access Journals (Sweden)

    Linley R. Schofield

    2015-01-01

    Full Text Available Pseudomurein endoisopeptidases cause lysis of the cell walls of methanogens by cleaving the isopeptide bond Ala-ε-Lys in the peptide chain of pseudomurein. PeiW and PeiP are two thermostable pseudomurein endoisopeptidases encoded by phage ΨM100 of Methanothermobacter wolfei and phages ΨM1 and ΨM2 of Methanothermobacter marburgensis, respectively. A continuous assay using synthetic peptide substrates was developed and used in the biochemical characterisation of recombinant PeiW and PeiP. The advantages of these synthetic peptide substrates over natural substrates are sensitivity, high purity, and characterisation and the fact that they are more easily obtained than natural substrates. In the presence of a reducing agent, purified PeiW and PeiP each showed similar activity under aerobic and anaerobic conditions. Both enzymes required a divalent metal for activity and showed greater thermostability in the presence of Ca2+. PeiW and PeiP involve a cysteine residue in catalysis and have a monomeric native conformation. The kinetic parameters, KM and kcat, were determined, and the ε-isopeptide bond between alanine and lysine was confirmed as the bond lysed by these enzymes in pseudomurein. The new assay may have wider applications for the general study of peptidases and the identification of specific methanogens susceptible to lysis by specific pseudomurein endoisopeptidases.

  20. Simultaneous Detection of Human C-Terminal p53 Isoforms by Single Template Molecularly Imprinted Polymers (MIPs) Coupled with Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)-Based Targeted Proteomics.

    Science.gov (United States)

    Jiang, Wenting; Liu, Liang; Chen, Yun

    2018-03-06

    Abnormal expression of C-terminal p53 isoforms α, β, and γ can cause the development of cancers including breast cancer. To date, much evidence has demonstrated that these isoforms can differentially regulate target genes and modulate their expression. Thus, quantification of individual isoforms may help to link clinical outcome to p53 status and to improve cancer patient treatment. However, there are few studies on accurate determination of p53 isoforms, probably due to sequence homology of these isoforms and also their low abundance. In this study, a targeted proteomics assay combining molecularly imprinted polymers (MIPs) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed for simultaneous quantification of C-terminal p53 isoforms. Isoform-specific surrogate peptides (i.e., KPLDGEYFTLQIR (peptide-α) for isoform α, KPLDGEYFTLQDQTSFQK (peptide-β) for isoform β, and KPLDGEYFTLQMLLDLR (peptide-γ) for isoform γ) were first selected and used in both MIPs enrichment and mass spectrometric detection. The common sequence KPLDGEYFTLQ of these three surrogate peptides was used as single template in MIPs. In addition to optimization of imprinting conditions and characterization of the prepared MIPs, binding affinity and cross-reactivity of the MIPs for each surrogate peptide were also evaluated. As a result, a LOQ of 5 nM was achieved, which was >15-fold more sensitive than that without MIPs. Finally, the assay was validated and applied to simultaneous quantitative analysis of C-terminal p53 isoforms α, β, and γ in several human breast cell lines (i.e., MCF-10A normal cells, MCF-7 and MDA-MB-231 cancer cells, and drug-resistant MCF-7/ADR cancer cells). This study is among the first to employ single template MIPs and cross-reactivity phenomenon to select isoform-specific surrogate peptides and enable simultaneous quantification of protein isoforms in LC-MS/MS-based targeted proteomics.

  1. Effects of saliva collection using cotton swabs on melatonin enzyme immunoassay.

    Science.gov (United States)

    Kozaki, Tomoaki; Lee, Soomin; Nishimura, Takayuki; Katsuura, Tetsuo; Yasukouchi, Akira

    2011-01-10

    Although various acceptable and easy-to-use devices have been used for saliva collection, cotton swabs are among the most common ones. Previous studies reported that cotton swabs yield a lower level of melatonin detection. However, this statistical method is not adequate for detecting an agreement between cotton saliva collection and passive saliva collection, and a test for bias is needed. Furthermore, the effects of cotton swabs have not been examined at lower melatonin level, a level at which melatonin is used for assessment of circadian rhythms, namely dim light melatonin onset (DLMO). In the present study, we estimated the effect of cotton swabs on the results of salivary melatonin assay using the Bland-Altman plot at lower level. Nine healthy males were recruited and each provided four saliva samples on a single day to yield a total of 36 samples. Saliva samples were directly collected in plastic tubes using plastic straws, and subsequently pipetted onto cotton swabs (cotton saliva collection) and into clear sterile tubes (passive saliva collection). The melatonin levels were analyzed in duplicate using commercially available ELISA kits. The mean melatonin concentration in cotton saliva collection samples was significantly lower than that in passive saliva collection samples at higher melatonin level (>6 pg/mL). The Bland-Altman plot indicated that cotton swabs causes relative and proportional biases in the assay results. For lower melatonin level (<6 pg/mL), although the BA plots didn't show proportional and relative biases, there was no significant correlation between passive and cotton saliva collection samples. Our findings indicate an interference effect of cotton swabs on the assay result of salivary melatonin at lower melatonin level. Cotton-based collection devices might, thus, not be suitable for assessment of DLMO.

  2. INFRARED STUDIES OF HUMAN SALIVA. IDENTIFICATION OF A FACTOR IN HUMAN SALIVA PRODUCING AN INFRARED ABSORBANCE MAXIMUM AT 4.9 MICRONS

    Science.gov (United States)

    An absorption maximum was observed at 4.9 microns in infrared spectra of human parotid saliva. The factor causing this absorbance was found to be a...nitrate, and heat stability. Thiocyanate was then determined in 16 parotid saliva samples by a spectrophotometric method, which involved formation of

  3. Immunization with 60 kD Ro peptide produces different stages of preclinical autoimmunity in a Sjögren's syndrome model among multiple strains of inbred mice.

    Science.gov (United States)

    Kurien, B T; Dsouza, A; Igoe, A; Lee, Y J; Maier-Moore, J S; Gordon, T; Jackson, M; Scofield, R H

    2013-07-01

    Sjögren's syndrome is a chronic illness manifested characteristically by immune injury to the salivary and lacrimal glands, resulting in dry mouth/eyes. Anti-Ro [Sjögren's syndrome antigen A (SSA)] and anti-La [Sjögren's syndrome antigen B (SSB)] autoantibodies are found frequently in Sjögren's subjects as well as in individuals who will go on to develop the disease. Immunization of BALB/c mice with Ro60 peptides results in epitope spreading with anti-Ro and anti-La along with lymphocyte infiltration of salivary glands similar to human Sjögren's. In addition, these animals have poor salivary function/low saliva volume. In this study, we examined whether Ro-peptide immunization produces a Sjögren's-like illness in other strains of mice. BALB/c, DBA-2, PL/J, SJL/J and C57BL/6 mice were immunized with Ro60 peptide-274. Sera from these mice were studied by immunoblot and enzyme-linked immunosorbent assay for autoantibodies. Timed salivary flow was determined after pharmacological stimulation, and salivary glands were examined pathologically. We found that SJL/J mice had no immune response to the peptide from Ro60, while C57BL/6 mice produced antibodies that bound the peptide but had no epitope spreading. PL/J mice had epitope spreading to other structures of Ro60 as well as to La, but like C57BL/6 and SJL/J had no salivary gland lymphocytic infiltration and no decrement of salivary function. DBA-2 and BALB/c mice had infiltration but only BALB/c had decreased salivary function. The immunological processes leading to a Sjögren's-like illness after Ro-peptide immunization were interrupted in a stepwise fashion in these differing mice strains. These data suggest that this is a model of preclinical disease with genetic control for epitope spreading, lymphocytic infiltration and glandular dysfunction. Published 2013. This article is a U.S. Government work and is in the public domain in the USA.

  4. Secretion of a recombinant protein without a signal peptide by the exocrine glands of transgenic rabbits.

    Directory of Open Access Journals (Sweden)

    Andrea Kerekes

    Full Text Available Transgenic rabbits carrying mammary gland specific gene constructs are extensively used for excreting recombinant proteins into the milk. Here, we report refined phenotyping of previously generated Venus transposon-carrying transgenic rabbits with particular emphasis on the secretion of the reporter protein by exocrine glands, such as mammary, salivary, tear and seminal glands. The Sleeping Beauty (SB transposon transgenic construct contains the Venus fluorophore cDNA, but without a signal peptide for the secretory pathway, driven by the ubiquitous CAGGS (CAG promoter. Despite the absence of a signal peptide, the fluorophore protein was readily detected in milk, tear, saliva and seminal fluids. The expression pattern was verified by Western blot analysis. Mammary gland epithelial cells of SB-CAG-Venus transgenic lactating does also showed Venus-specific expression by tissue histology and fluorescence microscopy. In summary, the SB-CAG-Venus transgenic rabbits secrete the recombinant protein by different glands. This finding has relevance not only for the understanding of the biological function of exocrine glands, but also for the design of constructs for expression of recombinant proteins in dairy animals.

  5. Secretion of a recombinant protein without a signal peptide by the exocrine glands of transgenic rabbits.

    Science.gov (United States)

    Kerekes, Andrea; Hoffmann, Orsolya Ivett; Iski, Gergely; Lipták, Nándor; Gócza, Elen; Kues, Wilfried A; Bősze, Zsuzsanna; Hiripi, László

    2017-01-01

    Transgenic rabbits carrying mammary gland specific gene constructs are extensively used for excreting recombinant proteins into the milk. Here, we report refined phenotyping of previously generated Venus transposon-carrying transgenic rabbits with particular emphasis on the secretion of the reporter protein by exocrine glands, such as mammary, salivary, tear and seminal glands. The Sleeping Beauty (SB) transposon transgenic construct contains the Venus fluorophore cDNA, but without a signal peptide for the secretory pathway, driven by the ubiquitous CAGGS (CAG) promoter. Despite the absence of a signal peptide, the fluorophore protein was readily detected in milk, tear, saliva and seminal fluids. The expression pattern was verified by Western blot analysis. Mammary gland epithelial cells of SB-CAG-Venus transgenic lactating does also showed Venus-specific expression by tissue histology and fluorescence microscopy. In summary, the SB-CAG-Venus transgenic rabbits secrete the recombinant protein by different glands. This finding has relevance not only for the understanding of the biological function of exocrine glands, but also for the design of constructs for expression of recombinant proteins in dairy animals.

  6. A synthetic peptide derived from alpha-fetoprotein inhibits the estradiol-induced proliferation of mammary tumor cells in culture through the modulation of p21.

    Science.gov (United States)

    Sierralta, Walter D; Epuñan, María J; Reyes, José M; Valladares, Luis E; Pino, Ana M

    2008-01-01

    A stable cyclized 9-mer peptide (cP) containing the active site of alpha-alpha fetoprotein (alphaFP) has been shown to be effective for prevention of estrogen-stimulated tumor cell proliferation in culture or of xenographt growth in immunodeficient mice. cP does not block 17beta-estradiol (E2) binding to its receptors, but rather appears to interfere with intracellular processing of the signal that supports growth. To obtain insight on that mechanism we studied the effect of cP on the proliferation of MCF-7 cells in culture. Proliferation in the presence of 2 microM E2 is decreased up to 40% upon addition of 2 microg ml(-1) cP to the medium; the presence of cP did not increase cell death, cP reduced also the proliferation of estrogen-dependent ZR75-1 cells but had no effect on autonomous MDA-MB-231 cells, cP did not modify the number of binding sites for labeled E2 or affected cell death. We detected increased nuclear p21Cip1 immunoreactivity after cP treatment. Our results suggest that cP acts via p21Cip1 to slow the process of MCF-7 cells through the cycle.

  7. Differentiation of oral bacteria in in vitro cultures and human saliva by secondary electrospray ionization - mass spectrometry

    Science.gov (United States)

    Bregy, Lukas; Müggler, Annick R.; Martinez-Lozano Sinues, Pablo; García-Gómez, Diego; Suter, Yannick; Belibasakis, Georgios N.; Kohler, Malcolm; Schmidlin, Patrick R.; Zenobi, Renato

    2015-10-01

    The detection of bacterial-specific volatile metabolites may be a valuable tool to predict infection. Here we applied a real-time mass spectrometric technique to investigate differences in volatile metabolic profiles of oral bacteria that cause periodontitis. We coupled a secondary electrospray ionization (SESI) source to a commercial high-resolution mass spectrometer to interrogate the headspace from bacterial cultures and human saliva. We identified 120 potential markers characteristic for periodontal pathogens Aggregatibacter actinomycetemcomitans (n = 13), Porphyromonas gingivalis (n = 70), Tanerella forsythia (n = 30) and Treponema denticola (n = 7) in in vitro cultures. In a second proof-of-principle phase, we found 18 (P. gingivalis, T. forsythia and T. denticola) of the 120 in vitro compounds in the saliva from a periodontitis patient with confirmed infection with P. gingivalis, T. forsythia and T. denticola with enhanced ion intensity compared to two healthy controls. In conclusion, this method has the ability to identify individual metabolites of microbial pathogens in a complex medium such as saliva.

  8. Effect of amino acid sequence and pH on nanofiber formation of self-assembling peptides EAK16-II and EAK16-IV.

    Science.gov (United States)

    Hong, Yooseong; Legge, Raymond L; Zhang, S; Chen, P

    2003-01-01

    Atomic force microscopy (AFM) and axisymmetric drop shape analysis-profile (ASDA-P) were used to investigate the mechanism of self-assembly of peptides. The peptides chosen consisted of 16 alternating hydrophobic and hydrophilic amino acids, where the hydrophilic residues possess alternating negative and positive charges. Two types of peptides, AEAEAKAKAEAEAKAK (EAK16-II) and AEAEAEAEAKAKAKAK (EAK16-IV), were investigated in terms of nanostructure formation through self-assembly. The experimental results, which focused on the effects of the amino acid sequence and pH, show that the nanostructures formed by the peptides are dependent on the amino acid sequence and the pH of the solution. For pH conditions around neutrality, one of the peptides used in this study, EAK16-IV, forms globular assemblies and has lower surface tension at air-water interfaces than another peptide, EAK16-II, which forms fibrillar assemblies at the same pH. When the pH is lowered below 6.5 or raised above 7.5, there is a transition from globular to fibrillar structures for EAK16-IV, but EAK16-II does not show any structural transition. Surface tension measurements using ADSA-P showed different surface activities of peptides at air-water interfaces. EAK16-II does not show a significant difference in surface tension for the pH range between 4 and 9. However, EAK16-IV shows a noticeable decrease in surface tension at pH around neutrality, indicating that the formation of globular assemblies is related to the molecular hydrophobicity.

  9. Cell phone use is associated with an inflammatory cytokine profile of parotid gland saliva.

    Science.gov (United States)

    Siqueira, Elisa Carvalho; de Souza, Fabrício Tinôco Alvim; Ferreira, Efigênia; Souza, Renan Pedra; Macedo, Samuel Costa; Friedman, Eitan; Gomez, Marcus Vinícius; Gomes, Carolina Cavaliéri; Gomez, Ricardo Santiago

    2016-10-01

    There is controversy on the effects of the non-ionizing radiation emitted by cell phones on cellular processes and the impact of such radiation exposure on health. The purpose of this study was to investigate whether cell phone use alters cytokine expression in the saliva produced by the parotid glands. Cytokine expression profile was determined by enzyme linked immuno sorbent assay (ELISA) in the saliva produced by the parotid glands in healthy volunteers, and correlated with self-reported cell phone use and laterality. The following parameters were determined, in 83 Brazilian individuals in saliva produced by the parotid glands comparing the saliva from the gland exposed to cell phone radiation (ipsilateral) to that from the contralateral parotid: salivary flow, total protein concentration, interleukin 1 β (IL-1 β), interleukin 6 (IL-6), interleukin 10 (IL-10), interferon γ (IFN-γ), and tumor necrosis factor α (TNF-α) salivary levels by ELISA. After multiple testing correction, decreased IL-10 and increased IL-1β salivary levels in the ipsilateral side compared with the contralateral side (P cell phones for more than 10 years presented higher differences between IL-10 levels in ipsilateral versus contralateral parotids (P = 0.0012). No difference was observed in any of the tested parameters in correlation with cell phone monthly usage in minutes. The exposure of parotid glands to cell phones can alter salivary IL-10 and IL-1β levels, consistent with a pro-inflammatory microenvironment that may be related to heat production. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  10. Interleukin-32 levels in gingival crevicular fluid and saliva of patients with chronic periodontitis after periodontal treatment.

    Science.gov (United States)

    Öngöz Dede, F; Balli, U; Bozkurt Doğan, Ş; Güven, B

    2017-06-01

    The cytokine, interleukin (IL)-32, is a relatively new discovery. However, it is very powerful for stimulating tumor necrosis factor-alpha (TNF-α) under inflammatory conditions. The objective of this research was to explore fluctuations in the levels of TNF-α, IL-32 and IL-10, in both saliva and gingival crevicular fluid. The focus was on measurements taken before and after clinical treatment of chronic periodontitis. For the purposes of the study, a total of 27 patients with chronic periodontitis and 27 controls (periodontally healthy) were recruited. Important clinical periodontal criteria were established before and 4 wk after the start of the research. The chronic periodontitis group was given an initial form of periodontal care. Samples of saliva and gingival crevicular fluid were collected exactly 4 wk preceding and 4 wk following the care. The levels of IL-10, IL-32 and TNF-α present in saliva and gingival crevicular fluid were recorded via the use of an ELISA. At baseline, the levels of TNF-α and IL-32 in the gingival crevicular fluid and saliva were significantly higher among patients in the chronic periodontitis group than among patients in the control group (p chronic periodontitis group than the control group (p chronic periodontitis group when compared with the baseline readings. However, the levels of IL-10 were significantly higher (p periodontitis. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  11. A distributive peptide cyclase processes multiple microviridin core peptides within a single polypeptide substrate.

    Science.gov (United States)

    Zhang, Yi; Li, Kunhua; Yang, Guang; McBride, Joshua L; Bruner, Steven D; Ding, Yousong

    2018-05-03

    Ribosomally synthesized and post-translationally modified peptides (RiPPs) are an important family of natural products. Their biosynthesis follows a common scheme in which the leader peptide of a precursor peptide guides the modifications of a single core peptide. Here we describe biochemical studies of the processing of multiple core peptides within a precursor peptide, rare in RiPP biosynthesis. In a cyanobacterial microviridin pathway, an ATP-grasp ligase, AMdnC, installs up to two macrolactones on each of the three core peptides within AMdnA. The enzyme catalysis occurs in a distributive fashion and follows an unstrict N-to-C overall directionality, but a strict order in macrolactonizing each core peptide. Furthermore, AMdnC is catalytically versatile to process unnatural substrates carrying one to four core peptides, and kinetic studies provide insights into its catalytic properties. Collectively, our results reveal a distinct biosynthetic logic of RiPPs, opening up the possibility of modular production via synthetic biology approaches.

  12. Translocation of cell-penetrating peptides into Candida fungal pathogens.

    Science.gov (United States)

    Gong, Zifan; Karlsson, Amy J

    2017-09-01

    Cell-penetrating peptides (CPPs) are small peptides capable of crossing cellular membranes while carrying molecular cargo. Although they have been widely studied for their ability to translocate nucleic acids, small molecules, and proteins into mammalian cells, studies of their interaction with fungal cells are limited. In this work, we evaluated the translocation of eleven fluorescently labeled peptides into the important human fungal pathogens Candida albicans and C. glabrata and explored the mechanisms of translocation. Seven of these peptides (cecropin B, penetratin, pVEC, MAP, SynB, (KFF) 3 K, and MPG) exhibited substantial translocation (>80% of cells) into both species in a concentration-dependent manner, and an additional peptide (TP-10) exhibiting strong translocation into only C. glabrata. Vacuoles were involved in translocation and intracellular trafficking of the peptides in the fungal cells and, for some peptides, escape from the vacuoles and localization in the cytosol were correlated to toxicity toward the fungal cells. Endocytosis was involved in the translocation of cecropin B, MAP, SynB, MPG, (KFF) 3 K, and TP-10, and cecropin B, penetratin, pVEC, and MAP caused membrane permeabilization during translocation. These results indicate the involvement of multiple translocation mechanisms for some CPPs. Although high levels of translocation were typically associated with toxicity of the peptides toward the fungal cells, SynB was translocated efficiently into Candida cells at concentrations that led to minimal toxicity. Our work highlights the potential of CPPs in delivering antifungal molecules and other bioactive cargo to Candida pathogens. © 2017 The Protein Society.

  13. Enzymatic characterization of peptidic materials isolated from aqueous solutions of ammonium cyanide (pH 9) and hydrocyanic acid (pH 6) exposed to ionizing radiation.

    Science.gov (United States)

    Niketic, V; Draganić, Z; Nesković, S; Draganić, I

    1982-01-01

    The enzymatic digestion of some radiolytically produced peptidic materials was examined. The substrates were compounds isolated from 0.1 molar solutions of NH4CN (pH 9) and HCN (pH 6), after their exposure to gamma rays from a 60Co source (15-20 Mrad doses). Commercial proteolytic enzymes pronase and aminopeptidase M were used. The examined materials were of composite nature and proteolytic action was systematically observed after their subsequent purification. In some fractions the effect was found to be positive with up to 30% of peptide bonds cleaved with respect to the amino acid content. These findings support our previous conclusions on the free radical induced formation of peptidic backbones without the intervention of amino acids. Some side effects were also noted which might be of interest in observations on enzymatic cleavage of other composite peptidic materials of abiotic origin.

  14. Characteristic changes of saliva and taste in burning mouth syndrome patients.

    Science.gov (United States)

    Imura, Hiroko; Shimada, Masahiko; Yamazaki, Yoko; Sugimoto, Kumiko

    2016-03-01

    Burning mouth syndrome (BMS) is characterized by chronic pain with a burning sensation of the tongue and oral mucosa and reported to be often accompanied by subjective xerostomia and dysgeusia. Since the etiology of BMS has not been elucidated, to understand the characteristics of BMS, we measured some components of saliva and taste sensitivity and compared the measured values between BMS and healthy subjects. Unstimulated saliva was collected from 15 female BMS patients and 30 healthy women. The flow rate, viscosity (spinnability) and concentration of secretory IgA (SIgA) of saliva and serum antioxidant capacity were measured. The recognition thresholds for sweet, salty, sour, bitter, and umami tastes were measured by whole-mouth method. The statistical analyses were performed using Student's t-test, and P < 0.05 was considered to be significant. In BMS group, the flow rate of saliva was significantly lower and the spinnability was significantly higher compared with healthy group. The secreted amount of SIgA per min and serum antioxidant capacity was significantly lower in the patients. The threshold for sourness in patients was significantly higher, while those for other tastes did not differ from healthy group. BMS patients showed lower salivary flow and higher salivary spinnability. These results together with decreased SIgA amount, suggest that BMS may be relevant to the deterioration of salivary condition, which could in turn affect taste function. Furthermore, the lower antioxidant capacity in patient's serum suggests that it can serve as a diagnostic tool for BMS. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  15. Identification of constrained peptides that bind to and preferentially inhibit the activity of the hepatitis C viral RNA-dependent RNA polymerase

    International Nuclear Information System (INIS)

    Amin, Anthony; Zaccardi, Joe; Mullen, Stanley; Olland, Stephane; Orlowski, Mark; Feld, Boris; Labonte, Patrick; Mak, Paul

    2003-01-01

    A class of disulfide constrained peptides containing a core motif FPWG was identified from a screen of phage displayed library using the HCV RNA-dependent RNA polymerase (NS5B) as a bait. Surface plasmon resonance studies showed that three highly purified synthetic constrained peptides bound to immobilized NS5B with estimated K d values ranging from 30 to 60 μM. In addition, these peptides inhibited the NS5B activity in vitro with IC 50 ranging from 6 to 48 μM, whereas in contrast they had no inhibitory effect on the enzymatic activities of calf thymus polymerase α, human polymerase β, RSV polymerase, and HIV reverse transcriptase in vitro. Two peptides demonstrated conformation-dependent inhibition since their synthetic linear versions were not inhibitory in the NS5B assay. A constrained peptide with the minimum core motif FPWG retained selective inhibition of NS5B activity with an IC 50 of 50 μM. Alanine scan analyses of a representative constrained peptide, FPWGNTW, indicated that residues F1 and W7 were critical for the inhibitory effect of this peptide, although residues P2 and N5 had some measurable inhibitory effect as well. Further analyses of the mechanism of inhibition indicated that these peptides inhibited the formation of preelongation complexes required for the elongation reaction. However, once the preelongation complex was formed, its activity was refractory to peptide inhibition. Furthermore, the constrained peptide FPWGNTW inhibited de novo initiated RNA synthesis by NS5B from a poly(rC) template. These data indicate that the peptides confer selective inhibition of NS5B activity by binding to the enzyme and perturbing an early step preceding the processive elongation step of RNA synthesis

  16. Significance of measurement of serum fibrosis markers (HA, LN, P III P, IV C) in patients with pulmonary tuberculosis

    International Nuclear Information System (INIS)

    Zhou Mingxian

    2006-01-01

    Objective: To study the clinical significance of determination of serum fibrosis markers in patients with pulmonary tuberculosis. Methods: Serum hyaluronic acid (HA), laminin (LN), procollagen III peptide (P III P) and Type IV collagen (IV C) contents were determined with RIA in 128 patients with pulmonary tuberculosis both before and after treatment as well as in 40 controls. Results: Before treatment, the serum contents of the four markers in the patients were significantly higher than those in the controls (P 0.05). Decreases of the levels in treatment failures were not significant and the levels remained significantly higher than those in controls (P<0.01 or P<0.05). Conclusion: Determination of serum fibrosis markers levels in patients with pulmonary tuberculosis is valuable for early diagnosis as well as for evaluation of the effect of chemotherapy. (authors)

  17. Myeloperoxidase activity is increased in gingival crevicular fluid and whole saliva after fixed orthodontic appliance activation.

    Science.gov (United States)

    Marcaccini, Andrea M; Amato, Patricia A F; Leão, Fernanda V; Gerlach, Raquel F; Ferreira, Jose T L

    2010-11-01

    Orthodontic tooth movement uses mechanical forces that result in inflammation in the first days. Myeloperoxidase (MPO) is an enzyme found in polymorphonuclear neutrophil (PMN) granules, and it is used to estimate the number of PMN granules in tissues. So far, MPO has not been used to study the inflammatory alterations after the application of orthodontic tooth movement forces. The aim of this study was to determine MPO activity in the gingival crevicular fluid (GCF) and saliva (whole stimulated saliva) of orthodontic patients at different time points after fixed appliance activation. MPO was determined in the GCF and collected by means of periopaper from the saliva of 14 patients with orthodontic fixed appliances. GCF and saliva samples were collected at baseline, 2 hours, and 7 and 14 days after application of the orthodontic force. Mean MPO activity was increased in both the GCF and saliva of orthodontic patients at 2 hours after appliance activation (P orthodontic force probably results in the increased MPO level observed at this time point. MPO might be a good marker to assess inflammation in orthodontic movement; it deserves further studies in orthodontic therapy. Copyright © 2010 American Association of Orthodontists. Published by Mosby, Inc. All rights reserved.

  18. Clinical significance of determination of changes of serum IL-6, hs-CRP and saliva secretory IgA levels after treatment in patients with periodontitis

    International Nuclear Information System (INIS)

    Wang Tongwu

    2009-01-01

    Objective: To explore the clinical significance of changes of serum IL-6, hs-CRP and saliva secretory IgA levels after treatment in patients with periodontitis. Methods: Serum IL-6, saliva secretory IgA (with RIA) and serum hs-CRP (with immuno-tarbility method) levels were measured in 42 patients with periodontitis both before and after treatment as well as in 35 controls. Results: Before treatment serum IL-6, hs-CRP and saliva secretory IgA levels in the patients wree significantly higher than those in controls (P 0.05). However, the saliva secreatory IgA levels were still significantly higher than those in controls (P<0.05). Conclusion: There was disturbance of immunomodulation in patients with periodontitis as expressed by the changes of cytokines levels in the course of the diseases. (authors)

  19. Effects of saliva collection using cotton swabs on melatonin enzyme immunoassay

    Directory of Open Access Journals (Sweden)

    Katsuura Tetsuo

    2011-01-01

    Full Text Available Abstract Background Although various acceptable and easy-to-use devices have been used for saliva collection, cotton swabs are among the most common ones. Previous studies reported that cotton swabs yield a lower level of melatonin detection. However, this statistical method is not adequate for detecting an agreement between cotton saliva collection and passive saliva collection, and a test for bias is needed. Furthermore, the effects of cotton swabs have not been examined at lower melatonin level, a level at which melatonin is used for assessment of circadian rhythms, namely dim light melatonin onset (DLMO. In the present study, we estimated the effect of cotton swabs on the results of salivary melatonin assay using the Bland-Altman plot at lower level. Methods Nine healthy males were recruited and each provided four saliva samples on a single day to yield a total of 36 samples. Saliva samples were directly collected in plastic tubes using plastic straws, and subsequently pipetted onto cotton swabs (cotton saliva collection and into clear sterile tubes (passive saliva collection. The melatonin levels were analyzed in duplicate using commercially available ELISA kits. Results The mean melatonin concentration in cotton saliva collection samples was significantly lower than that in passive saliva collection samples at higher melatonin level (>6 pg/mL. The Bland-Altman plot indicated that cotton swabs causes relative and proportional biases in the assay results. For lower melatonin level ( Conclusion Our findings indicate an interference effect of cotton swabs on the assay result of salivary melatonin at lower melatonin level. Cotton-based collection devices might, thus, not be suitable for assessment of DLMO.

  20. Shear Bond Strength of Orthodontic Brackets and Disinclusion Buttons: Effect of Water and Saliva Contamination

    Science.gov (United States)

    Sfondrini, Maria Francesca; Fraticelli, Danilo; Gandini, Paola

    2013-01-01

    Purpose. The aim of this study was to assess the effect of water and saliva contamination on the shear bond strength and failure site of orthodontic brackets and lingual buttons. Materials and Methods. 120 bovine permanent mandibular incisors were randomly divided into 6 groups of 20 specimens each. Both orthodontic brackets and disinclusion buttons were tested under three different enamel surface conditions: (a) dry, (b) water contamination, and (c) saliva contamination. Brackets and buttons were bonded to the teeth and subsequently tested using a Instron universal testing machine. Shear bond strength values and adhesive failure rate were recorded. Statistical analysis was performed using ANOVA and Tukey tests (strength values) and Chi squared test (ARI Scores). Results. Noncontaminated enamel surfaces showed the highest bond strengths for both brackets and buttons. Under water and saliva contamination orthodontic brackets groups showed significantly lower shear strengths than disinclusion buttons groups. Significant differences in debond locations were found among the groups under the various enamel surface conditions. Conclusions. Water and saliva contamination of enamel during the bonding procedure lowers bond strength values, more with orthodontic brackets than with disinclusion buttons. PMID:23762825