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Sample records for saliva dna yields

  1. DNA yield and quality of saliva samples and suitability for large-scale epidemiological studies in children.

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    Koni, A C; Scott, R A; Wang, G; Bailey, M E S; Peplies, J; Bammann, K; Pitsiladis, Y P

    2011-04-01

    To evaluate two saliva collection methods for DNA yield and quality as applied to a large, integrated, multicentre, European project involving the collection of biological material from children. Cross-sectional multicentre comparative study in young children. Saliva samples were collected from 14,019 children aged 2-9 years from eight European countries participating in the IDEFICS (Identification and prevention of dietary- and lifestyle-induced health effects in children and infants) study. This involved either the collection of 2 ml of saliva from children who were able to spit, or using a sponge to collect whole saliva and buccal mucosal cells from the inside of the mouth of younger children unable to spit. Samples were assembled centrally in each participating centre and subsequently despatched for DNA extraction and biobanking to the University of Glasgow. A subgroup of 4678 samples (∼33% of sampled individuals) were chosen for DNA extraction before genotyping. The whole-saliva collection method resulted in a higher DNA yield than the sponge collection method (mean±s.d.; saliva: 20.95±2.35 μg, sponge: 9.13±2.25 μg; Psaliva collection method compared with the assisted sponge collection. However, both collection methods provided DNA of sufficient quantity and quality for large-scale genetic epidemiological studies.

  2. Collection and extraction of saliva DNA for next generation sequencing.

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    Goode, Michael R; Cheong, Soo Yeon; Li, Ning; Ray, William C; Bartlett, Christopher W

    2014-08-27

    The preferred source of DNA in human genetics research is blood, or cell lines derived from blood, as these sources yield large quantities of high quality DNA. However, DNA extraction from saliva can yield high quality DNA with little to no degradation/fragmentation that is suitable for a variety of DNA assays without the expense of a phlebotomist and can even be acquired through the mail. However, at present, no saliva DNA collection/extraction protocols for next generation sequencing have been presented in the literature. This protocol optimizes parameters of saliva collection/storage and DNA extraction to be of sufficient quality and quantity for DNA assays with the highest standards, including microarray genotyping and next generation sequencing.

  3. The effect of age on DNA concentration from whole saliva: implications for the standard isolation method.

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    Gassó, Patricia; Pagerols, Mireia; Flamarique, Itziar; Castro-Fornieles, Josefina; Rodriguez, Natalia; Mas, Sergi; Curran, Sarah; Aitchison, Katherine; Santosh, Paramala; Lafuente, Amalia

    2014-01-01

    Adequate quantity and quality of the DNA isolated from saliva samples are crucial for ensuring successful genotyping rates in genetic studies. However, there is little information about these issues when saliva samples are collected from children. The objectives of this study were to assess whether there are differences in DNA quality or quantity isolated from saliva samples of children at different ages and adolescents compared to adults and, if so, to establish a modified protocol to improve and standardize DNA isolation from saliva samples of children. Saliva samples were collected with Oragene DNA Sample Collection Kit from 41 healthy subjects including children of different ages, adolescents, and adults. Quantity and quality of isolated DNA were determined spectrophotometrically. DNA concentration and age were positively correlated (r = 0.676, P children below 12 years yielded DNA concentrations saliva samples. This fact should be taken into account for a better standardization of the DNA isolation to ensure DNA banking in large-scale genetic studies involving children. © 2014 Wiley Periodicals, Inc.

  4. Saliva DNA quality and genotyping efficiency in a predominantly elderly population.

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    Gudiseva, Harini V; Hansen, Mark; Gutierrez, Linda; Collins, David W; He, Jie; Verkuil, Lana D; Danford, Ian D; Sagaser, Anna; Bowman, Anita S; Salowe, Rebecca; Sankar, Prithvi S; Miller-Ellis, Eydie; Lehman, Amanda; O'Brien, Joan M

    2016-04-07

    The question of whether DNA obtained from saliva is an acceptable alternative to DNA from blood is a topic of considerable interest for large genetics studies. We compared the yields, quality and performance of DNAs from saliva and blood from a mostly elderly study population. Two thousand nine hundred ten DNAs from primarily elderly subjects (mean age ± standard deviation (SD): 65 ± 12 years), collected for the Primary Open-Angle African-American Glaucoma Genetics (POAAGG) study, were evaluated by fluorometry and/or spectroscopy. These included 566 DNAs from blood and 2344 from saliva. Subsets of these were evaluated by Sanger sequencing (n = 1555), and by microarray SNP genotyping (n = 94) on an Illumina OmniExpress bead chip platform. The mean age of subjects was 65, and 68 % were female in both the blood and saliva groups. The mean ± SD of DNA yield per ml of requested specimen was significantly higher for saliva (17.6 ± 17.8 μg/ml) than blood (13.2 ± 8.5 μg/ml), but the mean ± SD of total DNA yield obtained per saliva specimen (35 ± 36 μg from 2 ml maximum specimen volume) was approximately three-fold lower than from blood (106 ± 68 μg from 8 ml maximum specimen volume). The average genotyping call rates were >99 % for 43 of 44 saliva DNAs and >99 % for 50 of 50 for blood DNAs. For 22 of 23 paired blood and saliva samples from the same individuals, the average genotyping concordance rate was 99.996 %. High quality PCR Sanger sequencing was obtained from ≥ 98 % of blood (n = 297) and saliva (n = 1258) DNAs. DNA concentrations ≥10 ng/μl, corresponding to total yields ≥ 2 μg, were obtained for 94 % of the saliva specimens (n = 2344). In spite of inferior purity, the performance of saliva DNAs for microarray genotyping was excellent. Our results agree with other studies concluding that saliva collection is a viable alternative to blood. The potential to boost study enrollments and reduce subject discomfort is not necessarily offset by a

  5. Saliva sampling in global clinical studies: the impact of low sampling volume on performance of DNA in downstream genotyping experiments

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    2013-01-01

    Background The collection of viable DNA samples is an essential element of any genetics research programme. Biological samples for DNA purification are now routinely collected in many studies with a variety of sampling methods available. Initial observation in this study suggested a reduced genotyping success rate of some saliva derived DNA samples when compared to blood derived DNA samples prompting further investigation. Methods Genotyping success rate was investigated to assess the suitability of using saliva samples in future safety and efficacy pharmacogenetics experiments. The Oragene® OG-300 DNA Self-Collection kit was used to collect and extract DNA from saliva from 1468 subjects enrolled in global clinical studies. Statistical analysis evaluated the impact of saliva sample volume of collection on the quality, yield, concentration and performance of saliva DNA in genotyping assays. Results Across 13 global clinical studies that utilized the Oragene® OG-300 DNA Self-Collection kit there was variability in the volume of saliva sample collection with ~31% of participants providing 0.5 mL of saliva, rather than the recommended 2 mL. While the majority of saliva DNA samples provided high quality genotype data, collection of 0.5 mL volumes of saliva contributed to DNA samples being significantly less likely to pass genotyping quality control standards. Assessment of DNA sample characteristics that may influence genotyping outcomes indicated that saliva sample volume, DNA purity and turbidity were independently associated with sample genotype pass rate, but that saliva collection volume had the greatest effect. Conclusion When employing saliva sampling to obtain DNA, it is important to encourage all study participants to provide sufficient sample to minimize potential loss of data in downstream genotyping experiments. PMID:23759220

  6. Assessing genetic polymorphisms using DNA extracted from cells present in saliva samples

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    Nemoda Zsofia

    2011-12-01

    Full Text Available Abstract Background Technical advances following the Human Genome Project revealed that high-quality and -quantity DNA may be obtained from whole saliva samples. However, usability of previously collected samples and the effects of environmental conditions on the samples during collection have not been assessed in detail. In five studies we document the effects of sample volume, handling and storage conditions, type of collection device, and oral sampling location, on quantity, quality, and genetic assessment of DNA extracted from cells present in saliva. Methods Saliva samples were collected from ten adults in each study. Saliva volumes from .10-1.0 ml, different saliva collection devices, sampling locations in the mouth, room temperature storage, and multiple freeze-thaw cycles were tested. One representative single nucleotide polymorphism (SNP in the catechol-0-methyltransferase gene (COMT rs4680 and one representative variable number of tandem repeats (VNTR in the serotonin transporter gene (5-HTTLPR: serotonin transporter linked polymorphic region were selected for genetic analyses. Results The smallest tested whole saliva volume of .10 ml yielded, on average, 1.43 ± .77 μg DNA and gave accurate genotype calls in both genetic analyses. The usage of collection devices reduced the amount of DNA extracted from the saliva filtrates compared to the whole saliva sample, as 54-92% of the DNA was retained on the device. An "adhered cell" extraction enabled recovery of this DNA and provided good quality and quantity DNA. The DNA from both the saliva filtrates and the adhered cell recovery provided accurate genotype calls. The effects of storage at room temperature (up to 5 days, repeated freeze-thaw cycles (up to 6 cycles, and oral sampling location on DNA extraction and on genetic analysis from saliva were negligible. Conclusions Whole saliva samples with volumes of at least .10 ml were sufficient to extract good quality and quantity DNA. Using

  7. Assessing genetic polymorphisms using DNA extracted from cells present in saliva samples

    Science.gov (United States)

    2011-01-01

    Background Technical advances following the Human Genome Project revealed that high-quality and -quantity DNA may be obtained from whole saliva samples. However, usability of previously collected samples and the effects of environmental conditions on the samples during collection have not been assessed in detail. In five studies we document the effects of sample volume, handling and storage conditions, type of collection device, and oral sampling location, on quantity, quality, and genetic assessment of DNA extracted from cells present in saliva. Methods Saliva samples were collected from ten adults in each study. Saliva volumes from .10-1.0 ml, different saliva collection devices, sampling locations in the mouth, room temperature storage, and multiple freeze-thaw cycles were tested. One representative single nucleotide polymorphism (SNP) in the catechol-0-methyltransferase gene (COMT rs4680) and one representative variable number of tandem repeats (VNTR) in the serotonin transporter gene (5-HTTLPR: serotonin transporter linked polymorphic region) were selected for genetic analyses. Results The smallest tested whole saliva volume of .10 ml yielded, on average, 1.43 ± .77 μg DNA and gave accurate genotype calls in both genetic analyses. The usage of collection devices reduced the amount of DNA extracted from the saliva filtrates compared to the whole saliva sample, as 54-92% of the DNA was retained on the device. An "adhered cell" extraction enabled recovery of this DNA and provided good quality and quantity DNA. The DNA from both the saliva filtrates and the adhered cell recovery provided accurate genotype calls. The effects of storage at room temperature (up to 5 days), repeated freeze-thaw cycles (up to 6 cycles), and oral sampling location on DNA extraction and on genetic analysis from saliva were negligible. Conclusions Whole saliva samples with volumes of at least .10 ml were sufficient to extract good quality and quantity DNA. Using 10 ng of DNA per

  8. Quality of DNA extracted from saliva samples collected with the Oragene™ DNA self-collection kit

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    Nunes Ana P

    2012-05-01

    Full Text Available Abstract Background Large epidemiological studies in DNA biobanks have increasingly used less invasive methods for obtaining DNA samples, such as saliva collection. Although lower amounts of DNA are obtained as compared with blood collection, this method has been widely used because of its more simple logistics and increased response rate. The present study aimed to verify whether a storage time of 8 months decreases the quality of DNA from collected samples. Methods Saliva samples were collected with an OrageneTM DNA Self-Collection Kit from 4,110 subjects aged 14–15 years. The samples were processed in two aliquots with an 8-month interval between them. Quantitative and qualitative evaluations were carried out in 20% of the samples by spectrophotometry and genotyping. Descriptive analyses and paired t-tests were performed. Results The mean volume of saliva collected was 2.2 mL per subject, yielding on average 184.8 μg DNA per kit. Most samples showed a Ratio of OD differences (RAT between 1.6 and 1.8 in the qualitative evaluation. The evaluation of DNA quality by TaqMan®, High Resolution Melting (HRM, and restriction fragment length polymorphism-PCR (RFLP-PCR showed a rate of success of up to 98% of the samples. The sample store time did not reduce either the quantity or quality of DNA extracted with the Oragene kit. Conclusion The study results showed that a storage period of 8 months at room temperature did not reduce the quality of the DNA obtained. In addition, the use of the Oragene kit during fieldwork in large population-based studies allows for DNA of high quantity and high quality.

  9. Clinical trial participant characteristics and saliva and DNA metrics

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    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.

  10. Environmental DNA from Residual Saliva for Efficient Noninvasive Genetic Monitoring of Brown Bears (Ursus arctos.

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    Rachel E Wheat

    Full Text Available Noninvasive genetic sampling is an important tool in wildlife ecology and management, typically relying on hair snaring or scat sampling techniques, but hair snaring is labor and cost intensive, and scats yield relatively low quality DNA. New approaches utilizing environmental DNA (eDNA may provide supplementary, cost-effective tools for noninvasive genetic sampling. We tested whether eDNA from residual saliva on partially-consumed Pacific salmon (Oncorhynchus spp. carcasses might yield suitable DNA quality for noninvasive monitoring of brown bears (Ursus arctos. We compared the efficiency of monitoring brown bear populations using both fecal DNA and salivary eDNA collected from partially-consumed salmon carcasses in Southeast Alaska. We swabbed a range of tissue types from 156 partially-consumed salmon carcasses from a midseason run of lakeshore-spawning sockeye (O. nerka and a late season run of stream-spawning chum (O. keta salmon in 2014. We also swabbed a total of 272 scats from the same locations. Saliva swabs collected from the braincases of salmon had the best amplification rate, followed by swabs taken from individual bite holes. Saliva collected from salmon carcasses identified unique individuals more quickly and required much less labor to locate than scat samples. Salmon carcass swabbing is a promising method to aid in efficient and affordable monitoring of bear populations, and suggests that the swabbing of food remains or consumed baits from other animals may be an additional cost-effective and valuable tool in the study of the ecology and population biology of many elusive and/or wide-ranging species.

  11. Environmental DNA from Residual Saliva for Efficient Noninvasive Genetic Monitoring of Brown Bears (Ursus arctos).

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    Wheat, Rachel E; Allen, Jennifer M; Miller, Sophie D L; Wilmers, Christopher C; Levi, Taal

    2016-01-01

    Noninvasive genetic sampling is an important tool in wildlife ecology and management, typically relying on hair snaring or scat sampling techniques, but hair snaring is labor and cost intensive, and scats yield relatively low quality DNA. New approaches utilizing environmental DNA (eDNA) may provide supplementary, cost-effective tools for noninvasive genetic sampling. We tested whether eDNA from residual saliva on partially-consumed Pacific salmon (Oncorhynchus spp.) carcasses might yield suitable DNA quality for noninvasive monitoring of brown bears (Ursus arctos). We compared the efficiency of monitoring brown bear populations using both fecal DNA and salivary eDNA collected from partially-consumed salmon carcasses in Southeast Alaska. We swabbed a range of tissue types from 156 partially-consumed salmon carcasses from a midseason run of lakeshore-spawning sockeye (O. nerka) and a late season run of stream-spawning chum (O. keta) salmon in 2014. We also swabbed a total of 272 scats from the same locations. Saliva swabs collected from the braincases of salmon had the best amplification rate, followed by swabs taken from individual bite holes. Saliva collected from salmon carcasses identified unique individuals more quickly and required much less labor to locate than scat samples. Salmon carcass swabbing is a promising method to aid in efficient and affordable monitoring of bear populations, and suggests that the swabbing of food remains or consumed baits from other animals may be an additional cost-effective and valuable tool in the study of the ecology and population biology of many elusive and/or wide-ranging species.

  12. DNA extraction from human saliva deposited on skin and its use in forensic identification procedures.

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    Anzai-Kanto, Evelyn; Hirata, Mário Hiroyuki; Hirata, Rosario Dominguez Crespo; Nunes, Fabio Daumas; Melani, Rodolfo Francisco Haltenhoff; Oliveira, Rogério Nogueira

    2005-01-01

    Saliva is usually deposited in bite marks found in many homicides, assault and other criminal cases. In the present study, saliva obtained from volunteers was deposited on skin and recovered for DNA extraction and typing in order to evaluate its usefulness for practical case investigation and discuss the contribution of forensic dentistry to saliva DNA typing. Twenty saliva samples were collected from different donors and used as suspects' samples. Five of these samples were randomly selected and deposited (250 microl) on arm skin. Saliva was collected from skin using the double swab technique. DNA from saliva and skin-deposited saliva samples was extracted by the phenol-chloroform method. DNA samples were amplified by PCR for DNA typing using a set of 15 STRs. The recovery of DNA from saliva deposited in the skin was 14 to 10 times lower than DNA quantity from saliva samples. DNA typing was demonstrated in 4 of 5 deposited saliva samples, the likelihood ratios estimated for these samples based on data of the Brazilian population were 1:11, 1:500, 1:159.140 and 1:153.700.123. Our results indicate that standardized procedures used for DNA collection and extraction from skin-deposited saliva can be used as a method to recover salivary DNA in criminal cases. However, it is important to observe that DNA recovery in forensic samples can be difficult. This study suggests that the analysis of saliva deposited on skin be incorporated into a criminal investigation since it may have great discriminatory power.

  13. HEPATITIS B VIRUS DNA IN SALIVA FROM CHILDREN WITH CHRONIC HEPATITIS B INFECTION IMPLICATIONS FOR SALIVA AS A POTENTIAL MODE OF HORIZONTAL TRANSMISSION

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    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

  14. 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...

  15. 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....

  16. Hepatitis B virus DNA in saliva from children with chronic hepatitis B infection: implications for saliva as a potential mode of horizontal transmission.

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    Heiberg, Ida Louise; Hoegh, Mette; Ladelund, Steen; Niesters, Hubert G M; Hogh, Birthe

    2010-05-01

    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.

  17. Search for varicella zoster virus DNA in saliva of healthy individuals aged 20-59 years.

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    Birlea, Marius; Cohrs, Randall J; Bos, Nathan; Mehta, Satish K; Pierson, Duane L; Gilden, Don

    2014-02-01

    All neurological and ocular complications of varicella zoster virus (VZV) reactivation can occur without rash. Virological verification requires detection of VZV DNA or anti-VZV IgG antibody in cerebrospinal fluid (CSF), or anti-VZV IgM antibody in serum or CSF. If VZV were readily detected in other tissue in patients with neurological disease without rash and found to correlate with tests listed above, more invasive tests such as lumbar puncture might be obviated. Saliva is a potential source of VZV DNA. To study the potential diagnostic value of detecting VZV DNA in saliva from patients with neurological disease, saliva of healthy adults was searched for VZV DNA. A single saliva sample obtained by passive drool was centrifuged at 16,000g for 20 min. DNA was extracted from the supernatant and cell pellet and examined in triplicate for VZV DNA by real time PCR. A single random saliva sample from 80 healthy men and women aged 20-59 years revealed no VZV DNA (Table ), but was uniformly positive for cell (GAPdH) DNA. Because VZV DNA was not found in a random saliva sample from 80 individuals 20-59-year-old, a VZV-positive sample during neurologic disease may have potential significance. Further studies will determine whether VZV DNA in saliva correlates with VZV DNA or anti-VZV antibody in CSF in patients with neurological disease. © 2013 Wiley Periodicals, Inc.

  18. Whole-Genome saliva and blood DNA methylation profiling in individuals with a respiratory allergy

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    Langie, Sabine A. S.; Szic, Katarzyna Szarc vel; Declerck, Ken

    2016-01-01

    development. High compliance rates can be expected in these studies when data is collected using non-invasive and convenient procedures. Saliva is an attractive biofluid to analyze changes in DNA methylation patterns. We investigated in a pilot study the differential methylation in saliva of RA (n = 5...

  19. Quantitative and qualitative assessment of DNA extracted from saliva for its use in forensic identification.

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    Khare, Parul; Raj, Vineet; Chandra, Shaleen; Agarwal, Suraksha

    2014-05-01

    Saliva has long been known for its diagnostic value in several diseases. It also has a potential to be used in forensic science. The objective of this study is to compare the quantity and quality of DNA samples extracted from saliva with those extracted from blood in order to assess the feasibility of extracting sufficient DNA from saliva for its possible use in forensic identification. Blood and saliva samples were collected from 20 volunteers and DNA extraction was performed through Phenol Chloroform technique. The quantity and quality of isolated DNA was analyzed by spectrophotometery and the samples were then used to amplify short tandem repeat (STR) F13 using the polymerase chain reaction. Mean quantity of DNA obtained in saliva was 48.4 ± 8.2 μg/ml and in blood was 142.5 ± 45.9 μg/ml. Purity of DNA obtained as assessed by the ratio of optical density 260/280, was found to be optimal in 45% salivary samples while remaining showed minor contamination. Despite this positive F13 STR amplification was achieved in 75% of salivary DNA samples. Results of this study showed that saliva may prove to be a useful source of DNA for forensic purpose.

  20. Whole-Genome Saliva and Blood DNA Methylation Profiling in Individuals with a Respiratory Allergy.

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    Sabine A S Langie

    Full Text Available The etiology of respiratory allergies (RA can be partly explained by DNA methylation changes caused by adverse environmental and lifestyle factors experienced early in life. Longitudinal, prospective studies can aid in the unravelment of the epigenetic mechanisms involved in the disease development. High compliance rates can be expected in these studies when data is collected using non-invasive and convenient procedures. Saliva is an attractive biofluid to analyze changes in DNA methylation patterns. We investigated in a pilot study the differential methylation in saliva of RA (n = 5 compared to healthy controls (n = 5 using the Illumina Methylation 450K BeadChip platform. We evaluated the results against the results obtained in mononuclear blood cells from the same individuals. Differences in methylation patterns from saliva and mononuclear blood cells were clearly distinguishable (PAdj0.2, though the methylation status of about 96% of the cg-sites was comparable between peripheral blood mononuclear cells and saliva. When comparing RA cases with healthy controls, the number of differentially methylated sites (DMS in saliva and blood were 485 and 437 (P0.1, respectively, of which 216 were in common. The methylation levels of these sites were significantly correlated between blood and saliva. The absolute levels of methylation in blood and saliva were confirmed for 3 selected DMS in the PM20D1, STK32C, and FGFR2 genes using pyrosequencing analysis. The differential methylation could only be confirmed for DMS in PM20D1 and STK32C genes in saliva. We show that saliva can be used for genome-wide methylation analysis and that it is possible to identify DMS when comparing RA cases and healthy controls. The results were replicated in blood cells of the same individuals and confirmed by pyrosequencing analysis. This study provides proof-of-concept for the applicability of saliva-based whole-genome methylation analysis in the field of respiratory allergy.

  1. Detection of Streptococcus mutans Genomic DNA in Human DNA Samples Extracted from Saliva and Blood

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    Vieira, Alexandre R.; Deeley, Kathleen B.; Callahan, Nicholas F.; Noel, Jacqueline B.; Anjomshoaa, Ida; Carricato, Wendy M.; Schulhof, Louise P.; DeSensi, Rebecca S.; Gandhi, Pooja; Resick, Judith M.; Brandon, Carla A.; Rozhon, Christopher; Patir, Asli; Yildirim, Mine; Poletta, Fernando A.; Mereb, Juan C.; Letra, Ariadne; Menezes, Renato; Wendell, Steven; Lopez-Camelo, Jorge S.; Castilla, Eduardo E.; Orioli, Iêda M.; Seymen, Figen; Weyant, Robert J.; Crout, Richard; McNeil, Daniel W.; Modesto, Adriana; Marazita, Mary L.

    2011-01-01

    Caries is a multifactorial disease, and studies aiming to unravel the factors modulating its etiology must consider all known predisposing factors. One major factor is bacterial colonization, and Streptococcus mutans is the main microorganism associated with the initiation of the disease. In our studies, we have access to DNA samples extracted from human saliva and blood. In this report, we tested a real-time PCR assay developed to detect copies of genomic DNA from Streptococcus mutans in 1,424 DNA samples from humans. Our results suggest that we can determine the presence of genomic DNA copies of Streptococcus mutans in both DNA samples from caries-free and caries-affected individuals. However, we were not able to detect the presence of genomic DNA copies of Streptococcus mutans in any DNA samples extracted from peripheral blood, which suggests the assay may not be sensitive enough for this goal. Values of the threshold cycle of the real-time PCR reaction correlate with higher levels of caries experience in children, but this correlation could not be detected for adults. PMID:21731912

  2. Flagellin enhances saliva IgA response and protection of anti-caries DNA vaccine.

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    Shi, W; Li, Y H; Liu, F; Yang, J Y; Zhou, D H; Chen, Y Q; Zhang, Y; Yang, Y; He, B X; Han, C; Fan, M W; Yan, H M

    2012-03-01

    We and others have shown that anti-caries DNA vaccines, including pGJA-P/VAX, are promising for preventing dental caries. However, challenges remain because of the low immunogenicity of DNA vaccines. In this study, we used recombinant flagellin protein derived from Salmonella (FliC) as a mucosal adjuvant for anti-caries DNA vaccine (pGJA-P/VAX) and analyzed the effects of FliC protein on the serum PAc-specific IgG and saliva PAc-specific IgA antibody responses, the colonization of Streptococcus mutans (S. mutans) on rat teeth, and the formation of caries lesions. Our results showed that FliC promoted the production of PAc-specific IgG in serum and secretory IgA (S-IgA) in saliva of rats by intranasal immunization with pGJA-P/VAX plus FliC. Furthermore, we found that enhanced PAc-specific IgA responses in saliva were associated with the inhibition of S. mutans colonization of tooth surfaces and endowed better protection with significant fewer caries lesions. In conclusion, our study demonstrates that recombinant FliC could enhance specific IgA responses in saliva and protective ability of pGJA-P/VAX, providing an effective mucosal adjuvant candidate for intranasal immunization of an anti-caries DNA vaccine.

  3. Direct PCR amplification of DNA from human bloodstains, saliva, and touch samples collected with microFLOQ®swabs.

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    Ambers, Angie; Wiley, Rachel; Novroski, Nicole; Budowle, Bruce

    2018-01-01

    Previous studies have shown that nylon flocked swabs outperform traditional fiber swabs in DNA recovery due to their innovative design and lack of internal absorbent core to entrap cellular materials. The microFLOQ ® Direct swab, a miniaturized version of the 4N6 FLOQSwab ® , has a small swab head that is treated with a lysing agent which allows for direct amplification and DNA profiling from sample collection to final result in less than two hours. Additionally, the microFLOQ ® system subsamples only a minute portion of a stain and preserves the vast majority of the sample for subsequent testing or re-analysis, if desired. The efficacy of direct amplification of DNA from dilute bloodstains, saliva stains, and touch samples was evaluated using microFLOQ ® Direct swabs and the GlobalFiler™ Express system. Comparisons were made to traditional methods to assess the robustness of this alternate workflow. Controlled studies with 1:19 and 1:99 dilutions of bloodstains and saliva stains consistently yielded higher STR peak heights than standard methods with 1ng input DNA from the same samples. Touch samples from common items yielded single source and mixed profiles that were consistent with primary users of the objects. With this novel methodology/workflow, no sample loss occurs and therefore more template DNA is available during amplification. This approach may have important implications for analysis of low quantity and/or degraded samples that plague forensic casework. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  4. Varicella-zoster DNA in saliva of patients with meningoencephalitis: a preliminary study.

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    Pollak, L; Mehta, S K; Pierson, D L; Sacagiu, T; Avneri Kalmanovich, S; Cohrs, R J

    2015-06-01

    Since the routine use of polymerase chain reaction testing (PCR) in diagnosing herpes infections, varicella-zoster virus is increasingly recognized as a cause of varicella-zoster meningoencephalitis (VZV ME) among immunocompetent patients. We were interested to determine whether patients with VZV ME had VZV DNA in their saliva during the acute phase of the illness. Forty-five consecutive patients who underwent a lumbar puncture for diagnostic purposes were included in the study. The cerebrospinal fluid was examined for the presence of VZV DNA by PCR, and patients with positive findings were treated with acyclovir. The saliva was later analyzed in a blinded fashion for the presence of VZV DNA. VZV DNA was found in saliva in four of five (80%) patients with PCR confirmed VZV ME (sensitivity 0.8, specificity 0.84, and likelihood ratio 5). This was significantly more than in patients with non-zoster viral ME (0%, P = 0.009), parainfectious headache (12%, P = 0.03) and controls (9.5%, P = 0.007). In immunocompromised patients with systemic lymphoma and AIDS, VZV DNA was present at a similar rate (67%, P = 0.6). We have found VZV DNA in saliva of patients with PCR confirmed VZV ME at a higher proportion than in controls and patients with non-VZV viral ME. This finding might be of clinical importance, especially in immunocompetent individuals with suspected VZV ME where the results of genetic and immunological testing are not conclusive. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  5. Parkinson's disease is associated with DNA methylation levels in human blood and saliva.

    Science.gov (United States)

    Chuang, Yu-Hsuan; Paul, Kimberly C; Bronstein, Jeff M; Bordelon, Yvette; Horvath, Steve; Ritz, Beate

    2017-08-30

    Several articles suggest that DNA methylation levels in blood relate to Parkinson's disease (PD) but there is a need for a large-scale study that involves suitable population based controls. The purposes of the study were: (1) to study whether PD status is associated with DNA methylation levels in blood/saliva; (2) to study whether observed associations relate to blood cell types; and (3) to characterize genome-wide significant markers ("CpGs") and clusters of CpGs (co-methylation modules) in terms of biological pathways. In a population-based case control study of PD, we studied blood samples from 335 PD cases and 237 controls and saliva samples from another 128 cases and 131 controls. DNA methylation data were generated from over 486,000 CpGs using the Illumina Infinium array. We identified modules of CpGs (clusters) using weighted correlation network analysis (WGCNA). Our cross-sectional analysis of blood identified 82 genome-wide significant CpGs (including cg02489202 in LARS2 p = 8.3 × 10(-11) and cg04772575 in ABCB9 p = 4.3 × 10(-10)). Three out of six PD related co-methylation modules in blood were significantly enriched with immune system related genes. Our analysis of saliva identified five significant CpGs. PD-related CpGs are located near genes that relate to mitochondrial function, neuronal projection, cytoskeleton organization, systemic immune response, and iron handling. This study demonstrates that: (1) PD status has a profound association with DNA methylation levels in blood and saliva; and (2) the most significant PD-related changes reflect changes in blood cell composition. Overall, this study highlights the role of the immune system in PD etiology but future research will need to address the causal structure of these relationships.

  6. DNA extracted from saliva for methylation studies of psychiatric traits: evidence tissue specificity and relatedness to brain.

    Science.gov (United States)

    Smith, Alicia K; Kilaru, Varun; Klengel, Torsten; Mercer, Kristina B; Bradley, Bekh; Conneely, Karen N; Ressler, Kerry J; Binder, Elisabeth B

    2015-01-01

    DNA methylation has become increasingly recognized in the etiology of psychiatric disorders. Because brain tissue is not accessible in living humans, epigenetic studies are most often conducted in blood. Saliva is often collected for genotyping studies but is rarely used to examine DNA methylation because the proportion of epithelial cells and leukocytes varies extensively between individuals. The goal of this study was to evaluate whether saliva DNA is informative for studies of psychiatric disorders. DNA methylation (HumanMethylation450 BeadChip) was assessed in saliva and blood samples from 64 adult African Americans. Analyses were conducted using linear regression adjusted for appropriate covariates, including estimated cellular proportions. DNA methylation from brain tissues (cerebellum, frontal cortex, entorhinal cortex, and superior temporal gyrus) was obtained from a publically available dataset. Saliva and blood methylation was clearly distinguishable though there was positive correlation overall. There was little correlation in CpG sites within relevant candidate genes. Correlated CpG sites were more likely to occur in areas of low CpG density (i.e., CpG shores and open seas). There was more variability in CpG sites from saliva than blood, which may reflect its heterogeneity. Finally, DNA methylation in saliva appeared more similar to patterns from each of the brain regions examined overall than methylation in blood. Thus, this study provides a framework for using DNA methylation from saliva and suggests that DNA methylation of saliva may offer distinct opportunities for epidemiological and longitudinal studies of psychiatric traits. © 2014 Wiley Periodicals, Inc.

  7. Monitoring of human herpesviruses-6 and -7 DNA in saliva samples during the acute and convalescent phases of exanthem subitum.

    Science.gov (United States)

    Miyazaki, Yuki; Namba, Hikaru; Torigoe, Sadayoshi; Watanabe, Masahiro; Yamashita, Nobuko; Ogawa, Hirohito; Morishima, Tsuneo; Yamada, Masao

    2017-04-01

    The amounts of the DNAs of human herpesviruses-6 (HHV-6) and -7 (HHV-7) in saliva samples were monitored during the acute and convalescent phases of exanthem subitum (ES) to elucidate the kinetics of virus shedding after ES. A total of 247 saliva samples were collected from 17 children (5 males and 12 females: 8-31 months old at onset). The monitoring period ranged from 152 to 721 days after onset, and in 15 children it was longer than 1 year. Among the 17 cases, 16 were attributed to HHV-6B, while a single case was attributed to HHV-7. Detection rates and average amounts of HHV-6 DNA in saliva samples after ES attributed to HHV-6B were low in the acute phase, increased to the maximum in the convalescent phase at 3-7 months, and then decreased. In addition, to investigate the source of infection, saliva samples from the older siblings (age 3-9 years) and parents of ES patients and children with a history of ES were also examined. The detection rate of HHV-6 DNA in saliva samples from 3- to 9-year-old children was significantly higher than the rate in adult saliva samples. Taken together, these findings suggest that the saliva of children in the convalescent phase of ES might be a more likely source of HHV-6 infection than that of adults. J. Med. Virol. 89:696-702, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  8. DNA methylation profiling for a confirmatory test for blood, saliva, semen, vaginal fluid and menstrual blood.

    Science.gov (United States)

    Lee, Hwan Young; Jung, Sang-Eun; Lee, Eun Hee; Yang, Woo Ick; Shin, Kyoung-Jin

    2016-09-01

    The ability to predict the type of tissues or cells from molecular profiles of crime scene samples has important practical implications in forensics. A previously reported multiplex assay using DNA methylation markers could only discriminate between 4 types of body fluids: blood, saliva, semen, and the body fluid which originates from female reproductive organ. In the present study, we selected 15 menstrual blood-specific CpG marker candidates based on analysis of 12 genome-wide DNA methylation profiles of vaginal fluid and menstrual blood. The menstrual blood-specificity of the candidate markers was confirmed by comparison with HumanMethylation450 BeadChip array data obtained for 58 samples including 12 blood, 12 saliva, 12 semen, 3 vaginal fluid, and 19 skin epidermis samples. Among 15CpG marker candidates, 3 were located in the promoter region of the SLC26A10 gene, and 2 of them (cg09696411 and cg18069290) showed high menstrual blood specificity. DNA methylation at the 2CpG markers was further tested by targeted bisulfite sequencing of 461 additional samples including 49 blood, 52 saliva, 34 semen, 125 vaginal fluid, and 201 menstrual blood. Because the 2 markers showed menstrual blood-specific methylation patterns, we modified our previous multiplex methylation SNaPshot reaction to include these 2 markers. In addition, a blood marker cg01543184 with cross reactivity to semen was replaced with cg08792630, and a semen-specific unmethylation marker cg17621389 was removed. The resultant multiplex methylation SNaPshot allowed positive identification of blood, saliva, semen, vaginal fluid and menstrual blood using the 9CpG markers which show a methylation signal only in the target body fluids. Because of the complexity in cell composition, menstrual bloods produced DNA methylation profiles that vary with menstrual cycle and sample collection methods, which are expected to provide more insight into forensic menstrual blood test. Moreover, because the developed

  9. DNA methylation-based age prediction from saliva: High age predictability by combination of 7 CpG markers.

    Science.gov (United States)

    Hong, Sae Rom; Jung, Sang-Eun; Lee, Eun Hee; Shin, Kyoung-Jin; Yang, Woo Ick; Lee, Hwan Young

    2017-07-01

    DNA methylation is currently one of the most promising age-predictive biomarkers. Many studies have reported DNA methylation-based age predictive models, but most of these are based on DNA methylation patterns from blood. Only a few studies have examined age-predictive DNA patterns in saliva, which is one of the most frequently-encountered body fluids at crime scenes. In this study, we generated genome-wide DNA methylation profiles of saliva from 54 individuals and identified CpG markers that showed a high correlation between methylation and age. Because the age-associated marker candidates from saliva differed from those of blood, we investigated DNA methylation patterns of 6 age-associated CpG marker candidates (cg00481951, cg19671120, cg14361627, cg08928145, cg12757011, and cg07547549 of the SST, CNGA3, KLF14, TSSK6, TBR1, and SLC12A5 genes, respectively) in addition to a cell type-specific CpG marker (cg18384097 of the PTPN7 gene) in an independent set of saliva samples obtained from 226 individuals aged 18 to 65 years. Multiplex methylation SNaPshot reactions were used to generate the data. We then generated a linear regression model with age information and the methylation profile from the 113 training samples. The model exhibited a 94.5% correlation between predicted and chronological age with a mean absolute deviation (MAD) from chronological age of 3.13 years. In subsequent validation using 113 test samples, we also observed a high correlation between predicted and chronological age (Spearman's rho=0.952, MAD from chronological age=3.15years). The model composed of 7 selected CpG sites enabled age prediction in saliva with high accuracy, which will be useful in saliva analysis for investigative leads. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. RNA/DNA co-analysis from human saliva and semen stains--results of a third collaborative EDNAP exercise

    DEFF Research Database (Denmark)

    Haas, Claus; Hanson, E; Anjos, M J

    2013-01-01

    A third collaborative exercise on RNA/DNA co-analysis for body fluid identification and STR profiling was organized by the European DNA Profiling Group (EDNAP). Twenty saliva and semen stains, four dilution series (10-0.01 µl saliva, 5-0.01 µl semen) and, optionally, bona fide or mock casework...... samples of human or non-human origin were analyzed by 20 participating laboratories using an RNA extraction or RNA/DNA co-extraction method. Two novel mRNA multiplexes were used: a saliva triplex (HTN3, STATH and MUC7) and a semen pentaplex (PRM1, PRM2, PSA, SEMG1 and TGM4). The laboratories used......), but allowed an STR profile of the stain donor to be obtained as well. The method proved to be reproducible and sensitive, with as little as 0.05 µl saliva or semen, using different analysis strategies. Additionally, we demonstrated the ability to positively identify the presence of saliva and semen, as well...

  11. Successful use of saliva without DNA extraction for detection of macrolide-resistant Mycoplasma pneumoniae DNA in children using LNA probe-based real-time PCR.

    Science.gov (United States)

    Komatsu, Haruki; Tsunoda, Tomoyuki; Inui, Ayano; Sogo, Tsuyoshi; Fujisawa, Tomoo; Imura, Motoki; Tateno, Akihiko

    2013-12-01

    It is not known that saliva is useful to diagnose Mycoplasma pneumoniae (M. pneumoniae) infection by PCR. We evaluated prospectively whether crude saliva samples without the DNA extraction process were useful for the detection of M. pneumoniae DNA in a locked nucleic acid (LNA) probe-based real-time PCR assay. Fifty-one clinical specimens (29 sputum, 22 saliva) that were positive by conventional M. pneumoniae-specific PCR were evaluated in this study. We designed an LNA probe-based real-time PCR that could discriminate the mutant strain (A2063G mutation) from the wild-type strain. All the 51 samples treated with DNA extraction were positive using the LNA probe-based real-time PCR. The results of the real-time PCR with DNA extraction were consistent with the sequence analysis. Of the 51 samples without DNA extraction, on the other hand, 41 (80.4%) were positive by real-time PCR. Of 29 sputum samples without DNA extraction, 23 (79.3%) were positive by real-time PCR; of the 22 saliva samples without DNA extraction, 18 (81.8%) were positive by real-time PCR. There was a statistically significant difference in the amplified DNA levels with extraction between the direct real-time PCR-positive samples (mean ± SD, 7.5 ± 1.6 log copies/ml) and PCR-negative samples (4.2 ± 0.8 log copies/ml) (P Saliva was useful for a template for PCR as well as sputum. In addition, crude samples were useful for real-time PCR when the samples had medium or high DNA levels. However, samples with low DNA levels sometimes showed false-negative results in direct real-time PCR.

  12. DNA extraction from human saliva deposited on skin and its use in forensic identification procedures Extração de DNA de saliva humana depositada sobre a pele e sua aplicabilidade aos processos de identificação forense

    Directory of Open Access Journals (Sweden)

    Evelyn Anzai-Kanto

    2005-09-01

    Full Text Available Saliva is usually deposited in bite marks found in many homicides, assault and other criminal cases. In the present study, saliva obtained from volunteers was deposited on skin and recovered for DNA extraction and typing in order to evaluate its usefulness for practical case investigation and discuss the contribution of forensic dentistry to saliva DNA typing. Twenty saliva samples were colleted from different donors and used as suspects' samples. Five of these samples were randomly selected and deposited (250 µl on arm skin. Saliva was collected from skin using the double swab technique. DNA from saliva and skin-deposited saliva samples was extracted by the phenol-chloroform method. DNA samples were amplified by PCR for DNA typing using a set of 15 STRs. The recovery of DNA from saliva deposited in the skin was 14 to 10 times lower than DNA quantity from saliva samples. DNA typing was demonstrated in 4 of 5 deposited saliva samples, the likelihood ratios estimated for these samples based on data of the Brazilian population were 1:11, 1:500, 1:159.140 and 1:153.700.123. Our results indicate that standardized procedures used for DNA collection and extraction from skin-deposited saliva can be used as a method to recover salivary DNA in criminal cases. However, it is important to observe that DNA recovery in forensic samples can be difficult. This study suggests that the analysis of saliva deposited on skin be incorporated into a criminal investigation since it may have great discriminatory power.A saliva é usualmente depositada em marcas de mordida encontradas em homicídios, agressões e outros crimes. Neste estudo, a saliva obtida de voluntários foi depositada na pele, recuperada para extração e tipagem do DNA, para avaliação de sua utilização e sua contribuição na odontologia legal. Vinte amostras de saliva foram coletadas de diferentes doadores e utilizadas como amostras de suspeitos. Cinco dessas amostras foram sorteadas e

  13. EVALUATION OF HBV DNA LEVELS IN SALIVA IN SUBJECTS WITH DIFFERENT VIREMIA AS WELL AS DURING PEGINTERFERON α-2a THERAPY.

    Directory of Open Access Journals (Sweden)

    Assya Krasteva

    2013-08-01

    Full Text Available Hepatitis B virus is the most important infectious hazard in dental profession. Vectors of HBV infection are not only blood, but also saliva, nasopharyngeal secretions, crevicular fluid.The aim of the study was to evaluate serum and salivary HBV DNA levels in subjects with chronic infection and dynamics of HBV levels during the first 3 months of peginterferon α-2a therapy to be determined. Nineteen parallel samples were tested for HBV DNA by real-time PCR assay. All nineteen sera were positive for HBV DNA with levels ranging from 494 to 6 300 000 000 cp/ml. HBV DNA was detected in all saliva samples even in patients with very low viremia. HBV DNA levels in serum and saliva were quite similar in cases with serum HBV DNA < 10 000 cp/ml. Patients with viremia higher than 10 000 cp/ml had significantly lower HBV DNA levels in saliva.The presence of HBV DNA in saliva might not be only due to transudation or exudation of fluid containing virus from the general circulation into various body fluids. These facts clearly demonstrate the role of saliva in routes of HBV transmission. Our results confirm the possibility of dentist’s particiation in infection transmission and suggests that salivary analysis holds promise as a non-invasive approach to identify biomarkers for diseases.

  14. Using a commercially available DNA extraction kit to obtain high quality human genomic DNA suitable for PCR and genotyping from 11-year-old saliva saturated cotton spit wads

    Directory of Open Access Journals (Sweden)

    Hudziak James J

    2008-12-01

    Full Text Available Abstract Background We sought to describe the integrity of human genomic DNA extracted from saliva saturated cotton spit wads stored at -20°C for approximately 11 years. 783 spit wad samples were collected from an ADHD sample population (Vermont Family Study during 1996–2000. Human genomic DNA was extracted from the spit wads using a commercially available kit; QIAamp DNA Blood Midi Kit (Qiagen, Inc., Valencia, CA. with a few modifications. Results The resulting DNA yield was more than adequate for genetic analysis and ranged from approximately 1 μg to a total of 80 μg (mean 17.3 μgs ± 11.9 μgs. A260/A280 ratios for the human genomic DNA extracted from the spit wads was consistently within the generally acceptable values of 1.7–2.0, with the lowest purity being 1.70, and a mean value of 1.937 ± 0.226 for the 783 samples. The DNA also was suitable for PCR reactions as evidenced by the amplification of the serotonin-transporter-linked polymorphic region, 5HTTLPR. 5HTTLPR is a functional polymorphism in the promoter region of the serotonin transporter gene (HTT, SLC6A4, or SERT, consisting of two intensively studied alleles. 770 of the 783 samples (98.3% produced fragments after PCR of the expected size with primers specific for 5HTTLPR. Conclusion High quality and abundant genomic DNA can be successfully retrieved from saliva saturated cotton spit wads using the commercially available kit, QIAamp DNA Blood Midi Kit from Qiagen, Inc. Furthermore, the DNA can be extracted in less than 3 hours and multiple samples can be processed simultaneously thus reducing processing time.

  15. High yield DNA fragmentation using cyclical hydrodynamic shearing

    NARCIS (Netherlands)

    Shui, Lingling; Sparreboom, Wouter; Spang, Peter; Roeser, Tina; Nieto, Benjamin; Guasch, Francesc; Corbera, Antoni Homs; van den Berg, Albert; Carlen, Edwin

    2013-01-01

    We report a new DNA fragmentation technique that significantly simplifies conventional hydrodynamic shearing fragmentation by eliminating the need for sample recirculation while maintaining high fragmentation yield and low fragment length variation, and therefore, reduces instrument complexity and

  16. Paramagnetic Cellulose DNA Isolation Improves DNA Yield and Quality Among Diverse Plant Taxa

    Directory of Open Access Journals (Sweden)

    Jackson R. Moeller

    2014-10-01

    Full Text Available Premise of the study: The chemical diversity of land plants ensures that no single DNA isolation method results in high yield and purity with little effort for all species. Here we evaluate a new technique originally developed for forensic science, based on MagnaCel paramagnetic cellulose particles (PMC, to determine its efficacy in extracting DNA from 25 plant species representing 21 families and 15 orders. Methods and Results: Yield and purity of DNA isolated by PMC, DNeasy Plant Mini Kit (silica column, and cetyltrimethylammonium bromide (CTAB methods were compared among four individuals for each of 25 plant species. PMC gave a two-fold advantage in average yield, and the relative advantage of the PMC method was greatest for samples with the lowest DNA yields. PMC also produced more consistent sample purity based on absorbance ratios at 260 : 280 and 260 : 230 nm. Conclusions: PMC technology is a promising alternative for plant DNA isolation.

  17. Diagnostic Usefulness of Varicella-Zoster Virus Real-Time Polymerase Chain Reaction Analysis of DNA in Saliva and Plasma Specimens From Patients With Herpes Zoster.

    Science.gov (United States)

    Park, Seong Yeon; Kim, Ji Yeun; Kim, Ji-Ae; Kwon, Ji-Soo; Kim, Sun-Mi; Jeon, Na Young; Kim, Min-Chul; Chong, Yong Pil; Lee, Sang-Oh; Choi, Sang-Ho; Kim, Yang Soo; Woo, Jun Hee; Kim, Sung-Han

    2017-12-27

    We evaluated the diagnostic usefulness of polymerase chain reaction (PCR) analysis for detecting varicella-zoster virus (VZV) infection and reactivation of VZV, using DNA extracted from saliva and plasma specimens obtained from subjects with suspected herpes zoster and from healthy volunteers during stressful and nonstressful conditions. There were 52 patients with a diagnosis of herpes zoster (group 1), 30 with a diagnosis of zoster-mimicking disease (group 2), and 27 healthy volunteers (group 3). Saliva and plasma samples were evaluated for VZV DNA by real-time PCR analysis. Among patients with suspected herpes zoster (ie, patients in groups 1 and 2), the sensitivity of PCR analysis of salivary DNA for detecting VZV (88%; 95% confidence interval [CI], 74%-95%) was significantly higher than that of PCR analysis of plasma DNA (28%; 95% CI, 16%-44%; P .99). VZV DNA was not detected in saliva and plasma samples from group 3 (0%; 95% CI, 0%-14%). Real-time PCR analysis of salivary DNA is more sensitive than that of plasma DNA for detecting VZV among patients with suspected herpes zoster. We found no subclinical reactivation of VZV in group 3 following exposure to common stressful conditions.

  18. Sticky Saliva

    Science.gov (United States)

    McCarroll, Louise; Solomon, Michael; Schultz, William

    2016-11-01

    Oral and even systemic health begins with healthy saliva by maintaining antibacterial activity, lubricating hard and soft oral tissues, healing, tasting, chewing, and swallowing. Saliva functionality is intimately linked to its rheology. Alterations in saliva rheology may indicate or cause unhealthy biological function. One imprecise pathological designation is "sticky saliva", usually self-reported or qualitatively described by health professionals. Saliva is 99% water and therefore behaves like water in shear. Saliva also contains mucins, electrolytes, enzymes, hormones, and antibodies. These additional constituents enable saliva to form a long-lasting filament with a "beads-on-a-string" morphology in extension. Therefore, the main kinematic feature that distinguishes the coupling between the oral cavity and saliva elongational mechanics. We investigate the effect of pH and salinity on saliva filament formation with preliminary experiments and compare to 1D unsteady viscoelastic models. We discuss the results in the context of saliva functionality and in generating more satisfactory saliva substitutes for those suffering from xerostomia. We will discuss when beads-on-a-string are likely to occur.

  19. Epstein-Barr virus antibodies in serum and DNA load in saliva are not associated with radiological or clinical disease activity in patients with early multiple sclerosis.

    Directory of Open Access Journals (Sweden)

    René M Gieß

    Full Text Available To investigate the association of Epstein-Barr virus (EBV nuclear antigen-1 (EBNA-1 and viral capsid antigen (VCA immunoglobulin (IgG antibodies in serum as well as EBV DNA load in saliva with radiological and clinical disease activity in patients with clinically isolated syndrome (CIS and early relapsing-remitting MS (RRMS.EBNA-1 and VCA immunoglobulin (IgG antibodies were determined in serum of 100 patients with CIS/early RRMS and 60 healthy controls. EBV DNA load was measured in saliva of 48 patients and 50 controls. Patients underwent clinical assessment with the Expanded Disability Status Scale (EDSS and 3 Tesla magnetic resonance imaging at baseline and after a median of 20 months of follow-up (n = 63 for MRI, n = 71 for EDSS. The association of EBV parameters with occurrence of a second relapse, indicating conversion to clinically definite MS (CDMS, was evaluated over a median of 35 months of follow-up after the first clinical event (n = 89.EBNA-1 IgG antibody frequency (p = 0.00005 and EBNA-1 and VCA IgG antibody levels (p<0.0001 for both were higher in patients than in controls. EBV DNA load in saliva did not differ between groups. Neither EBV antibody levels nor DNA load in saliva were associated with baseline or follow-up number or volume of T2-weighted (T2w or contrast enhancing lesions, number of Barkhof criteria or the EDSS, or with the number of new T2w lesions, T2w lesion volume change or EDSS change on follow-up. Likewise, levels of EBV IgG antibodies in serum and DNA load in saliva were not associated with conversion to CDMS.While these findings confirm the association of EBV infection with early MS, neither EBNA-1 nor VCA IgG antibodies in serum nor EBV DNA load in saliva were associated with radiological or clinical disease activity in patients with CIS/early RRMS. These data are compatible with the concept that EBV may be a trigger for MS acting very early during the development of the disease.

  20. Comprehensive examination of conventional and innovative body fluid identification approaches and DNA profiling of laundered blood- and saliva-stained pieces of cloths.

    Science.gov (United States)

    Kulstein, G; Wiegand, P

    2018-01-01

    Body fluids like blood and saliva are commonly encountered during investigations of high volume crimes like homicides. The identification of the cellular origin and the composition of the trace can link suspects or victims to a certain crime scene and provide a probative value for criminal investigations. To erase all traces from the crime scene, perpetrators often wash away their traces. Characteristically, items that show exposed stains like blood are commonly cleaned or laundered to free them from potential visible leftovers. Mostly, investigators do not delegate the DNA analysis of laundered items. However, some studies have already revealed that items can still be used for DNA analysis even after they have been laundered. Nonetheless, a systematical evaluation of laundered blood and saliva traces that provides a comparison of different established and newly developed methods for body fluid identification (BFI) is still missing. Herein, we present the results of a comprehensive study of laundered blood- and saliva-stained pieces of cloths that were applied to a broad range of methods for BFI including conventional approaches as well as molecular mRNA profiling. The study included the evaluation of cellular origin as well as DNA profiling of blood- and saliva-stained (synthetic fiber and cotton) pieces of cloths, which have been washed at various washing temperatures for one or multiple times. Our experiments demonstrate that, while STR profiling seems to be sufficiently sensitive for the individualization of laundered items, there is a lack of approaches for BFI with the same sensitivity and specificity allowing to characterize the cellular origin of challenging, particularly laundered, blood and saliva samples.

  1. 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.

  2. Epstein-Barr virus antibodies in serum and DNA load in saliva are not associated with radiological or clinical disease activity in patients with early multiple sclerosis.

    Science.gov (United States)

    Gieß, René M; Pfuhl, Catherina; Behrens, Janina R; Rasche, Ludwig; Freitag, Erik; Khalighy, Nima; Otto, Carolin; Wuerfel, Jens; Brandt, Alexander U; Hofmann, Jörg; Eberspächer, Bettina; Bellmann-Strobl, Judith; Paul, Friedemann; Ruprecht, Klemens

    2017-01-01

    To investigate the association of Epstein-Barr virus (EBV) nuclear antigen-1 (EBNA-1) and viral capsid antigen (VCA) immunoglobulin (Ig)G antibodies in serum as well as EBV DNA load in saliva with radiological and clinical disease activity in patients with clinically isolated syndrome (CIS) and early relapsing-remitting MS (RRMS). EBNA-1 and VCA immunoglobulin (Ig)G antibodies were determined in serum of 100 patients with CIS/early RRMS and 60 healthy controls. EBV DNA load was measured in saliva of 48 patients and 50 controls. Patients underwent clinical assessment with the Expanded Disability Status Scale (EDSS) and 3 Tesla magnetic resonance imaging at baseline and after a median of 20 months of follow-up (n = 63 for MRI, n = 71 for EDSS). The association of EBV parameters with occurrence of a second relapse, indicating conversion to clinically definite MS (CDMS), was evaluated over a median of 35 months of follow-up after the first clinical event (n = 89). EBNA-1 IgG antibody frequency (p = 0.00005) and EBNA-1 and VCA IgG antibody levels (psaliva did not differ between groups. Neither EBV antibody levels nor DNA load in saliva were associated with baseline or follow-up number or volume of T2-weighted (T2w) or contrast enhancing lesions, number of Barkhof criteria or the EDSS, or with the number of new T2w lesions, T2w lesion volume change or EDSS change on follow-up. Likewise, levels of EBV IgG antibodies in serum and DNA load in saliva were not associated with conversion to CDMS. While these findings confirm the association of EBV infection with early MS, neither EBNA-1 nor VCA IgG antibodies in serum nor EBV DNA load in saliva were associated with radiological or clinical disease activity in patients with CIS/early RRMS. These data are compatible with the concept that EBV may be a trigger for MS acting very early during the development of the disease.

  3. High yield and high quality DNA from vegetative and sexual tissues ...

    African Journals Online (AJOL)

    Pines are considered to be difficult for DNA extraction. However, from one species to the other there is variation in phenolic profiles and seed size that might affect final DNA yields and quality. Two DNA extraction protocols (CTAB and SDS based) were compared for their ability to produce DNA on leaves, gametophyte and ...

  4. Differences in the quantity of DNA found in the urine and saliva of smokers versus nonsmokers: implications for the timing of epigenetic events.

    Science.gov (United States)

    Simkin, Melissa; Abdalla, Moemen; El-Mogy, Mohamed; Haj-Ahmad, Yousef

    2012-06-01

    TP53 is a tumor-suppressor gene coding for p53, a protein responsible for cell-cycle arrest and DNA repair. Smoking has been demonstrated to lead to the methylation of tumor-suppressor genes in noncancerous lung biopsy tissues of smokers, and in bodily fluids, promoter hypermethylation occurs very early in the progression of cancer. Thus, DNA methylation changes may be initiated long before cells become cancerous. As this association has never been explored in young, healthy individuals, we decided to look at DNA isolated from urine and saliva samples taken from young male and female smoking and nonsmoking participants. While p53 methylation was not found in any of the samples tested, differences in DNA concentration between the two groups may shed light on the timing of epigenetic alterations, as well as better explain why the negative impact of smoking is not often found in young, healthy adults.

  5. SDH Subunit Mutation Status in Saliva: Genetic Testing in Patients with Pheochromocytoma.

    Science.gov (United States)

    Osinga, T E; Xekouki, P; Nambuba, J; Faucz, F R; de la Luz Sierra, M; Links, T P; Kema, I P; Adams, K; Stratakis, C A; van der Horst-Schrivers, A N A; Pacak, K

    2016-04-01

    Germline mutations occur in up to 30-40% of pheochromocytoma/paraganglioma, with mutations in the succinate dehydrogenase (SDH) subunits B (SDHB) and D (SDHD) being the most common. Blood samples are favored for obtaining high quality DNA, however, leukocytes can also be obtained by collecting saliva. The aim of this study was to determine whether SDHB and SDHD gene mutations in patients with pheochromocytoma/paraganglioma could be determined using a salivary sample. Paired blood and salivary samples were collected from 30 patients: 9 SDHB mutation positive, 13 with a SDHD mutation, and 8 without any SDHx mutations. The Oragene DISCOVER kit was used to collect and extract DNA from saliva. Blood DNA was extracted from EDTA blood samples. The DNA purification and concentration were measured by spectrophotometry. The 8 exons of SDHB and the 4 exons of SDHD were amplified and sequenced by PCR-based bidirectional Sanger sequencing. Total DNA yields from blood DNA were similar to those obtained from saliva DNA [mean (±SD) saliva vs. blood DNA concentration 514.6 (±580.8) ng/µl vs. 360.9 (±262.7) ng/µl; p=0.2)]. The purity of the saliva DNA samples was lower than that of blood [mean OD260/OD280 ratio 1.78 (±0.13) vs. 1.87 (±0.04); p=0.001, respectively], indicating more protein contamination in the saliva-extracted DNA. This study shows that salivary DNA collected from patients with pheochromocytoma/paraganglioma is a good alternative for extraction of genomic DNA for its high DNA concentration and acceptable purity and can be used as an alternative to blood derived DNA in screening for SDHB and SDHD mutations. © Georg Thieme Verlag KG Stuttgart · New York.

  6. PCR detection of multiple human herpesvirus DNA in saliva from HIV-infected individuals in Teresina, State of Piauí, Brazil Detecção por PCR do DNA de vários herpesvírus humanos na saliva de indivíduos infectados pelo HIV em Teresina, Estado do Piauí, Brasil

    Directory of Open Access Journals (Sweden)

    Kátia Silene Sousa Carvalho

    2010-12-01

    Full Text Available INTRODUCTION: Human herpesviruses are frequently associated with orofacial diseases in humans (HSV-1, EBV, CMV and HHV-8, some can also cause systemic disease (CMV and HHV-8. The transmission of these viruses occurs by contact with infected secretions, especially saliva. Human immunodeficiency virus infection is associated with an increased risk of HHVs and related diseases. METHODS: This work aimed to detect HSV-1, EBV, CMV and HHV-8 DNA in saliva of HIV-infected patients from Teresina, northeast Brazil, by PCR and compare these findings with age and sex matched HIV-seronegative individuals. RESULTS: No difference in prevalence was verified between HHV detection in the saliva of HIV-seropositive individuals and controls. The individual frequencies of these viruses in these two populations were different. HIV seropositivity correlated positively with the presence of CMV (OR: 18.2, p= 0.00032 and EBV (OR: 3.44, p= 0.0081. No association between CD4 counts and the prevalence of HHVs in the saliva was observed; however, a strong association was determined between seropositivity and the presence of multiple HHV DNAs in saliva (OR: 4.83, p = 0.0028. CONCLUSIONS: These findings suggest the asymptomatic salivary shedding of HHVs is a common event between HIV-seropositive and seronegative individuals from Teresina, Piauí, Brazil, and, especially for HIV-seropositive patients, saliva is a risk factor for the acquisition/transmission of multiple HHVs.INTRODUÇÃO: Alguns herpesvírus humanos são frequentemente associados a doenças orofaciais em humanos. A transmissão destes vírus ocorre através do contato com secreções contaminadas, especialmente a saliva. A infecção pelo vírus da imunodeficiência humana é considerada um fator de risco para a aquisição de HHVs e doenças correlatas. MÉTODOS: Este trabalho teve como objetivo detectar por PCR o DNA de HSV-1, EBV, CMV e HHV-8 na saliva de pacientes infectados com HIV em Teresina, nordeste do

  7. Antipsychotic drugs attenuate aberrant DNA methylation of DTNBP1 (dysbindin) promoter in saliva and post-mortem brain of patients with schizophrenia and Psychotic bipolar disorder.

    Science.gov (United States)

    Abdolmaleky, Hamid M; Pajouhanfar, Sara; Faghankhani, Masoomeh; Joghataei, Mohammad Taghi; Mostafavi, Ashraf; Thiagalingam, Sam

    2015-12-01

    Due to the lack of genetic association between individual genes and schizophrenia (SCZ) pathogenesis, the current consensus is to consider both genetic and epigenetic alterations. Here, we report the examination of DNA methylation status of DTNBP1 promoter region, one of the most credible candidate genes affected in SCZ, assayed in saliva and post-mortem brain samples. The Illumina DNA methylation profiling and bisulfite sequencing of representative samples were used to identify methylation status of the DTNBP1 promoter region. Quantitative methylation specific PCR (qMSP) was employed to assess methylation of DTNBP1 promoter CpGs flanking a SP1 binding site in the saliva of SCZ patients, their first-degree relatives and control subjects (30, 15, and 30/group, respectively) as well as in post-mortem brains of patients with SCZ and bipolar disorder (BD) versus controls (35/group). qRT-PCR was used to assess DTNBP1 expression. We found DNA hypermethylation of DTNBP1 promoter in the saliva of SCZ patients (∼12.5%, P = 0.036), particularly in drug-naïve patients (∼20%, P = 0.011), and a trend toward hypermethylation in their first-degree relatives (P = 0.085) versus controls. Analysis of post-mortem brain samples revealed an inverse correlation between DTNBP1 methylation and expression, and normalization of this epigenetic change by classic antipsychotic drugs. Additionally, BD patients with psychotic depression exhibited higher degree of methylation versus other BD patients (∼80%, P = 0.025). DTNBP1 promoter DNA methylation may become a key element in a panel of biomarkers for diagnosis, prevention, or therapy in SCZ and at risk individuals pending confirmatory studies with larger sample sizes to attain a higher degree of significance. © 2015 Wiley Periodicals, Inc.

  8. Extracting DNA from 'jaws': High yield and quality from archived tiger shark (Galeocerdo cuvier) skeletal material

    DEFF Research Database (Denmark)

    Eg Nielsen, Einar; Morgan, J. A T; Maher, S. L.

    2017-01-01

    archived skeletal material from sharks as a source of DNA for temporal genomic studies. Six different methods for DNA extraction, encompassing two different commercial kits and three different protocols, were applied to material, so-called bio-swarf, from contemporary and archived jaws and vertebrae...... of tiger sharks (Galeocerdo cuvier). Protocols were compared for DNA yield and quality using a qPCR approach. For jaw swarf, all methods provided relatively high DNA yield and quality, while large differences in yield between protocols were observed for vertebrae. Similar results were obtained from samples...... of white shark (Carcharodon carcharias). Application of the optimized methods to 38 museum and private angler trophy specimens dating back to 1912 yielded sufficient DNA for downstream genomic analysis for 68% of the samples. No clear relationships between age of samples, DNA quality and quantity were...

  9. A high throughput DNA extraction method with high yield and quality

    Directory of Open Access Journals (Sweden)

    Xin Zhanguo

    2012-07-01

    Full Text Available Abstract Background Preparation of large quantity and high quality genomic DNA from a large number of plant samples is a major bottleneck for most genetic and genomic analyses, such as, genetic mapping, TILLING (Targeting Induced Local Lesion IN Genome, and next-generation sequencing directly from sheared genomic DNA. A variety of DNA preparation methods and commercial kits are available. However, they are either low throughput, low yield, or costly. Here, we describe a method for high throughput genomic DNA isolation from sorghum [Sorghum bicolor (L. Moench] leaves and dry seeds with high yield, high quality, and affordable cost. Results We developed a high throughput DNA isolation method by combining a high yield CTAB extraction method with an improved cleanup procedure based on MagAttract kit. The method yielded large quantity and high quality DNA from both lyophilized sorghum leaves and dry seeds. The DNA yield was improved by nearly 30 fold with 4 times less consumption of MagAttract beads. The method can also be used in other plant species, including cotton leaves and pine needles. Conclusion A high throughput system for DNA extraction from sorghum leaves and seeds was developed and validated. The main advantages of the method are low cost, high yield, high quality, and high throughput. One person can process two 96-well plates in a working day at a cost of $0.10 per sample of magnetic beads plus other consumables that other methods will also need.

  10. A high throughput DNA extraction method with high yield and quality.

    Science.gov (United States)

    Xin, Zhanguo; Chen, Junping

    2012-07-28

    Preparation of large quantity and high quality genomic DNA from a large number of plant samples is a major bottleneck for most genetic and genomic analyses, such as, genetic mapping, TILLING (Targeting Induced Local Lesion IN Genome), and next-generation sequencing directly from sheared genomic DNA. A variety of DNA preparation methods and commercial kits are available. However, they are either low throughput, low yield, or costly. Here, we describe a method for high throughput genomic DNA isolation from sorghum [Sorghum bicolor (L.) Moench] leaves and dry seeds with high yield, high quality, and affordable cost. We developed a high throughput DNA isolation method by combining a high yield CTAB extraction method with an improved cleanup procedure based on MagAttract kit. The method yielded large quantity and high quality DNA from both lyophilized sorghum leaves and dry seeds. The DNA yield was improved by nearly 30 fold with 4 times less consumption of MagAttract beads. The method can also be used in other plant species, including cotton leaves and pine needles. A high throughput system for DNA extraction from sorghum leaves and seeds was developed and validated. The main advantages of the method are low cost, high yield, high quality, and high throughput. One person can process two 96-well plates in a working day at a cost of $0.10 per sample of magnetic beads plus other consumables that other methods will also need.

  11. [Application of saliva in disease diagnosis].

    Science.gov (United States)

    Xingqun, Cheng; Xuedong, Zhou; Xin, Xu

    2016-12-01

    Saliva is secreted by salivary glands and performs a variety of functions, including mouth cleaning and protection, antibacterial activity, and digestion. With the rapid progress in salivaomics, saliva became recognized as a potential pool of biological markers. Being a non-invasive and safe source, saliva is a potential substitute for blood in diagnosis and prognosis of diseases. This review summarizes the latest advancement in saliva-related studies and presents the potential value of saliva in early diagnosis of oral diseases, such as dental caries, periodontal disease, cancer, diabetes, and other systemic disorders. Saliva biomarkers can reveal changes ranging from changes in biochemical index, DNA, RNA, and proteins to the diversification of microbiota structure. By integrating recent data, this paper discusses the clinical significance and application prospect of saliva in early diagnosis of diseases and in translational and precision medicine.

  12. A nanodosimetric model of radiation-induced clustered DNA damage yields

    Science.gov (United States)

    Garty, G.; Schulte, R.; Shchemelinin, S.; Leloup, C.; Assaf, G.; Breskin, A.; Chechik, R.; Bashkirov, V.; Milligan, J.; Grosswendt, B.

    2010-02-01

    We present a nanodosimetric model for predicting the yield of double strand breaks (DSBs) and non-DSB clustered damages induced in irradiated DNA. The model uses experimental ionization cluster size distributions measured in a gas model by an ion counting nanodosimeter or, alternatively, distributions simulated by a Monte Carlo track structure code developed to simulate the nanodosimeter. The model is based on a straightforward combinatorial approach translating ionizations, as measured or simulated in a sensitive gas volume, to lesions in a DNA segment of one-two helical turns considered equivalent to the sensitive volume of the nanodosimeter. The two model parameters, corresponding to the probability that a single ion detected by the nanodosimeter corresponds to a single strand break or a single lesion (strand break or base damage) in the equivalent DNA segment, were tuned by fitting the model-predicted yields to previously measured double-strand break and double-strand lesion yields in plasmid DNA irradiated with protons and helium nuclei. Model predictions were also compared to both yield data simulated by the PARTRAC code for protons of a wide range of different energies and experimental DSB and non-DSB clustered DNA damage yield data from the literature. The applicability and limitations of this model in predicting the LET dependence of clustered DNA damage yields are discussed.

  13. Kr-86 Ion-Beam Irradiation of Hydrated DNA: Free Radical and Unaltered Base Yields

    Science.gov (United States)

    Becker, David; Adhikary, Amitava; Tetteh, Smedley T.; Bull, Arthur W.; Sevilla, Michael D.

    2012-01-01

    This work reports an ESR and product analysis investigation of Kr-86 ion-beam irradiation of hydrated DNA at 77 K. The irradiation results in the formation and trapping of both base radicals and sugar phosphate radicals (DNA backbone radicals). The absolute yields (G, μmol/J) of the base radicals are smaller than the yields found in similarly prepared γ-irradiated DNA samples, and the relative yields of backbone radicals relative to base radicals are much higher than that found in γ-irradiated samples. From these results, we have elaborated our radiation chemical model of the track structure for ion-beam irradiated DNA as it applies to krypton ion-beams. The base radicals, which are trapped as ion radicals or reversibly protonated or deprotonated ion radicals, are formed almost entirely in the track penumbra, a region in which radiation chemical effects are similar to those found in γ-irradiated samples. By comparing the yields of base radicals in ion-beam samples to the yields of the same radicals in γ-irradiated samples, the partition of energy between the low-LET region (penumbra) and the core is experimentally determined. The neutral sugar and other backbone radicals, which are not as susceptible to recombination as are ion radicals, are formed largely in the track core. The backbone radicals show a linear dose response up to very high doses. Unaltered base release yields in Kr-86 irradiated hydrated DNA are equal to sugar radical yields within experimental error limits, consistent with radiation-chemical processes in which all base release originates with sugar radicals. Two phosphorus-centered radicals from fragmentation of the DNA backbone are found in low yields. PMID:23106211

  14. Simple, High-Yield Syntheses of DNA Duplexes Containing Interstrand DNA-DNA Cross-links Between an N4-Aminocytidine Residue and an Abasic Site

    OpenAIRE

    Gamboa Varela, Jacqueline; Gates, Kent S.

    2016-01-01

    The protocol describes the preparation and purification of interstrand DNA-DNA cross-links derived from the reaction of an N4-aminocytidine residue with an abasic site in duplex DNA. The procedures employ inexpensive, commercially-available chemicals and enzymes to carry out post-synthetic modification of commercially-available oligodeoxynucleotides. The yield of cross-linked duplex is typically better than 90%. If purification is required, the cross-linked duplex can be readily separated fro...

  15. Large-scale plasmid DNA processing: evidence that cell harvesting and storage methods affect yield of supercoiled plasmid DNA.

    Science.gov (United States)

    Kong, Simyee; Rock, Cassandra F; Booth, Andrew; Willoughby, Nicholas; O'Kennedy, Ronan D; Relton, Julian; Ward, John M; Hoare, Mike; Levy, M Susana

    2008-09-01

    The effect of bacterial-cell centrifugation and handling on the initial stages of plasmid processing was investigated. Escherichia coli cells containing either a 6 or 20 kb plasmid were grown in 75- and 450-litre bioreactors, and the process yield of the early recovery stages was characterized in terms of SC pDNA (supercoiled plasmid DNA) recovered. In all cases, the cells were totally recovered using either a continuous-feed, intermittent-solids-discharge, disc-stack centrifuge or a continuous-feed, batch-discharge, solid-bowl centrifuge. The cells were then either processed immediately or stored frozen. The centrifugation method considerably affected the yield of SC pDNA, and there was evidence that the intermittent discharge of cells from a centrifuge operating at high speed led to a sediment containing lysed cells and degraded pDNA. This led to estimated plasmid yield losses of up to 40% as compared with cells recovered from laboratory or solid-bowl centrifuges, where there is evidently no cell stress on discharge. By inference, the cell stress on feed to either of the continuous centrifuges studied was not implicated in product loss. Freezing of the recovered cells gives a convenient hold stage prior to further processing. In all cases, this extra freeze-thaw stage led to loss of SC pDNA, and this was in addition to the loss attributed to cell lysis during centrifugation discharge. Only average yields can be gained from pilot plant-scale studies; separate laboratory-based experiments indicated that this loss of SC pDNA is determined by the time and temperature for which the resuspended cells are held.

  16. SU-E-T-05: Comparing DNA Strand Break Yields for Photons under Different Irradiation Conditions with Geant4-DNA.

    Science.gov (United States)

    Pater, P; Bernal, M; Naqa, I El; Seuntjens, J

    2012-06-01

    To validate and scrutinize published DNA strand break data with Geant4-DNA and a probabilistic model. To study the impact of source size, electronic equilibrium and secondary electron tracking cutoff on direct relative biological effectiveness (DRBE). Geant4 (v4.9.5) was used to simulate a cylindrical region of interest (ROI) with r = 15 nm and length = 1.05 mm, in a slab of liquid water of 1.06 g/cm 3 density. The ROI was irradiated with mono-energetic photons, with a uniformly distributed volumetric isotropic source (0.28, 1.5 keV) or a plane beam (0.662, 1.25 MeV), of variable size. Electrons were tracked down to 50 or 10 eV, with G4-DNA processes and energy transfer greater than 10.79 eV was scored. Based on volume ratios, each scored event had a 0.0388 probability of happening on either DNA helix (break). Clusters of at least one break on each DNA helix within 3.4 nm were found using a DBSCAN algorithm and categorized as double strand breaks (DSB). All other events were categorized as single strand breaks (SSB). Geant4-DNA is able to reproduce strand break yields previously published. Homogeneous irradiation conditions should be present throughout the ROI for DRBE comparisons. SSB yields seem slightly dependent on the primary photon energy. DRBEs show a significant increasing trend for lower energy incident photons. A lower electron cutoff produces higher SSB yields, but decreases the SSB/DSB yields ratio. The probabilistic and geometrical DNA models can predict equivalent results. Using Geant4, we were able to reproduce previously published results on the direct strand break yields of photon and study the importance of irradiation conditions. We also show an ascending trend for DRBE with lower incident photon energies. A probabilistic model coupled with track structure analysis can be used to simulate strand break yields. NSERC, CIHR. © 2012 American Association of Physicists in Medicine.

  17. HLA polymorphisms and detection of kaposi sarcoma-associated herpesvirus DNA in saliva and peripheral blood among children and their mothers in the uganda sickle cell anemia KSHV Study

    Directory of Open Access Journals (Sweden)

    Figueiredo Constança

    2010-11-01

    Full Text Available Abstract Kaposi sarcoma-associated herpesvirus (KSHV, also called Human herpesvirus 8 or HHV8 is a γ-2 herpesvirus that causes Kaposi sarcoma. KSHV seroprevalence rates vary geographically with variable rates recorded in different sub Sahara African countries, suggesting that effects of genetic and/or environmental factors may influence the risk of infection. One study conducted in South Africa, where KSHV seroprevalence is relatively low, found that carriage of human leukocyte antigen (HLA alleles HLA-A*6801, HLA-A*30, HLA-A*4301, and HLA-DRB1*04 was associated with increased shedding of KSHV DNA in saliva. Confirmation of those results would strengthen the hypothesis that genetic factors may influence KSHV distribution by modulating KSHV shedding in saliva. To explore these associations in another setting, we used high resolution HLA-A, B, and DRB1 typing on residual samples from the Uganda Sickle Cell Anemia KSHV study, conducted in a high KSHV seroprevalence region, to investigate associations between HLA and KSHV shedding in saliva or peripheral blood among 233 children and their mothers. HLA-A and HLA-DRB1 alleles were not associated with KSHV shedding in our study, but our study was small and was not adequately powered to exclude small associations. In exploratory analyses, we found marginal association of KSHV DNA shedding in saliva but not in peripheral blood among children carrying HLA- B*4415 and marginal association of KSHV DNA shedding in peripheral blood but not in saliva among children carrying HLA- B*0801 alleles. The contribution of individual HLA polymorphisms to KSHV shedding is important but it may vary in different populations. Larger population-based studies are needed to estimate the magnitude and direction of association of HLA with KSHV shedding and viral control.

  18. Nanofabrication Yields. Hybridization and Click-Fixation of Polycyclic DNA Nanoassemblies

    KAUST Repository

    Lundberg, Erik P.

    2011-09-27

    We demonstrate the stepwise assembly of a fully addressable polycyclic DNA hexagon nanonetwork for the preparation of a four-ring system, one of the biggest networks yet constructed from tripodal building blocks. We find that the yield exhibits a distinct upper level <100%, a fundamental problem of thermodynamic DNA assembly that appears to have been overlooked in the DNA nanotechnology literature. A simplistic model based on a single step-yield parameter y can quantitatively describe the total yield of DNA assemblies in one-pot reactions as Y = yduplex n, with n the number of hybridization steps. Experimental errors introducing deviations from perfect stoichiometry and the thermodynamics of hybridization equilibria contribute to decreasing the value of yduplex (on average y = 0.96 for our 10 base pair hybridization). For the four-ring system (n = 31), the total yield is thus less than 30%, which is clearly unsatisfactory if bigger nanoconstructs of this class are to be designed. Therefore, we introduced site-specific click chemistry for making and purifying robust building blocks for future modular constructs of larger assemblies. Although the present yield of this robust module was only about 10%, it demonstrates a first step toward a general fabrication approach. Interestingly, we find that the click yields follow quantitatively a binomial distribution, the predictability of which indicates the usefulness of preparing pools of pure and robust building blocks in this way. The binomial behavior indicates that there is no interference between the six simultaneous click reactions but that step-yield limiting factors such as topological constraints and Cu(I) catalyst concentration are local and independent. © 2011 American Chemical Society.

  19. An evaluation of DNA yield, DNA quality and bite registration from a dental impression wafer.

    Science.gov (United States)

    Ellis, Mark A; Song, Fengyu; Parks, Edwin T; Eckert, George J; Dean, Jeffrey A; Windsor, L Jack

    2007-09-01

    The authors determined the amount and quality of the DNA captured by a bite impression wafer and analyzed any inaccuracies in the impression wafer. The authors made bite registrations for subjects aged 7 to 12 years by using a dental impression wafer (Toothprints, Kerr, Orange, Calif.), obtained an oral rinse sample, took cheek cells by using buccal swabs and made an alginate impression to pour a stone model. They extracted and quantified the DNA from the dental impression wafer, mouthwash and buccal swabs by using the Quant-iT PicoGreen (Invitrogen, Carlsbad, Calif.) assay and a real-time polymerase chain reaction (RT-PCR) assay. They compared the stone models and imprints from the wafer. The average amounts of DNA determined by using Quant-iT PicoGreen from the buccal swab, mouthwash and dental impression wafer samples were 113.61, 509.57 and 1.03 micrograms, respectively. The average amounts of DNA determined by using RT-PCR from the buccal swab, mouthwash and dental impression wafer samples were 11.5240, 22.2540 and 0.0279 mug, respectively. The bite registrations and stone models had an average of 14 percent of mismatches. The dental impression wafers captured DNA but not in high quantities. They did not produce an accurate representation of the dentition. The dental impression wafers captured enough DNA to permit amplification. The accuracy of the bite registration was not sufficient for identification purposes. Therefore, dental impression wafers may be useful only as a reservoir for DNA.

  20. Lutzomyia longipalpis Saliva or Salivary Protein LJM19 Protects against Leishmania braziliensis and the Saliva of Its Vector, Lutzomyia intermedia

    Science.gov (United States)

    Tavares, Natalia M.; Silva, Robson A.; Costa, Dirceu J.; Pitombo, Maiana A.; Fukutani, Kiyoshi F.; Miranda, José C.; Valenzuela, Jesus G.; Barral, Aldina; de Oliveira, Camila I.; Barral-Netto, Manoel; Brodskyn, Claudia

    2011-01-01

    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 transmitted by a different

  1. 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

  2. Proteome Analysis of Watery Saliva Secreted by Green Rice Leafhopper, Nephotettix cincticeps.

    Directory of Open Access Journals (Sweden)

    Makoto Hattori

    Full Text Available The green rice leafhopper, Nephotettix cincticeps, is a vascular bundle feeder that discharges watery and gelling saliva during the feeding process. To understand the potential functions of saliva for successful and safe feeding on host plants, we analyzed the complexity of proteinaceous components in the watery saliva of N. cincticeps. Salivary proteins were collected from a sucrose diet that adult leafhoppers had fed on through a membrane of stretched parafilm. Protein concentrates were separated using SDS-PAGE under reducing and non-reducing conditions. Six proteins were identified by a gas-phase protein sequencer and two proteins were identified using LC-MS/MS analysis with reference to expressed sequence tag (EST databases of this species. Full -length cDNAs encoding these major proteins were obtained by rapid amplification of cDNA ends-PCR (RACE-PCR and degenerate PCR. Furthermore, gel-free proteome analysis that was performed to cover the broad range of salivary proteins with reference to the latest RNA-sequencing data from the salivary gland of N. cincticeps, yielded 63 additional protein species. Out of 71 novel proteins identified from the watery saliva, about 60 % of those were enzymes or other functional proteins, including GH5 cellulase, transferrin, carbonic anhydrases, aminopeptidase, regucalcin, and apolipoprotein. The remaining proteins appeared to be unique and species- specific. This is the first study to identify and characterize the proteins in watery saliva of Auchenorrhyncha species, especially sheath-producing, vascular bundle-feeders.

  3. Saliva and wound healing

    NARCIS (Netherlands)

    Brand, H.S.; Veerman, E.C.I.

    2013-01-01

    Wounds in the oral cavity heal faster and with less scarring than wounds in other parts of the body. One of the factors implicated in this phenomenon is the presence of saliva, which promotes the healing of oral wounds in several ways. Saliva creates a humid environment, which improves the survival

  4. Saliva and wound healing

    NARCIS (Netherlands)

    Brand, H.S.; Ligtenberg, A.J.M.; Veerman, E.C.I.; Ligtenberg, A.J.M.; Veerman, E.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

  5. Factors influencing the yield of satellite DNA in extractions from Drosophila virilis and Drosophila melanogaster adults and embryos.

    Science.gov (United States)

    Rae, P M; Barnett, T R; Babbitt, D G

    1976-05-03

    The application of different DNA extraction methods to identical batches of Drosophila virilis and Drosophila melanogaster flies or embryos has revealed that the ionic strength of a homogenization medium is of critical importance if chloroform extractions are performed. The low yield of satellite DNA after homogenization in low salt buffers is less severe if EDTA is included in the buffer. Phenol extraction procedures result in no such differential behavior of satellite and main band DNA, but under certain circumstances a particular satellite fraction of Drosophila virilis DNA may be lost.

  6. 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.

  7. 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. 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. PMID:23138733

  8. Saliva diagnostics - Current views and directions.

    Science.gov (United States)

    Kaczor-Urbanowicz, Karolina Elżbieta; Martin Carreras-Presas, Carmen; Aro, Katri; Tu, Michael; Garcia-Godoy, Franklin; Wong, David Tw

    2017-03-01

    In this review, we provide an update on the current and future applications of saliva for diagnostic purposes. There are many advantages of using saliva as a biofluid. Its collection is fast, easy, inexpensive, and non-invasive. In addition, saliva, as a "mirror of the body," can reflect the physiological and pathological state of the body. Therefore, it serves as a diagnostic and monitoring tool in many fields of science such as medicine, dentistry, and pharmacotherapy. Introduced in 2008, the term "Salivaomics" aimed to highlight the rapid development of knowledge about various "omics" constituents of saliva, including: proteome, transcriptome, micro-RNA, metabolome, and microbiome. In the last few years, researchers have developed new technologies and validated a wide range of salivary biomarkers that will soon make the use of saliva a clinical reality. However, a great need still exists for convenient and accurate point-of-care devices that can serve as a non-invasive diagnostic tool. In addition, there is an urgent need to decipher the scientific rationale and mechanisms that convey systemic diseases to saliva. Another promising technology called liquid biopsy enables detection of circulating tumor cells (CTCs) and fragments of tumor DNA in saliva, thus enabling non-invasive early detection of various cancers. The newly developed technology-electric field-induced release and measurement (EFIRM) provides near perfect detection of actionable mutations in lung cancer patients. These recent advances widened the salivary diagnostic approach from the oral cavity to the whole physiological system, and thus point towards a promising future of salivary diagnostics for personalized individual medicine applications including clinical decisions and post-treatment outcome predictions. Impact statement The purpose of this mini-review is to make an update about the present and future applications of saliva as a diagnostic biofluid in many fields of science such as dentistry

  9. Saliva diagnostics – Current views and directions

    Science.gov (United States)

    Kaczor-Urbanowicz, Karolina Elżbieta; Martin Carreras-Presas, Carmen; Aro, Katri; Tu, Michael; Wong, David TW

    2016-01-01

    In this review, we provide an update on the current and future applications of saliva for diagnostic purposes. There are many advantages of using saliva as a biofluid. Its collection is fast, easy, inexpensive, and non-invasive. In addition, saliva, as a “mirror of the body,” can reflect the physiological and pathological state of the body. Therefore, it serves as a diagnostic and monitoring tool in many fields of science such as medicine, dentistry, and pharmacotherapy. Introduced in 2008, the term “Salivaomics” aimed to highlight the rapid development of knowledge about various “omics” constituents of saliva, including: proteome, transcriptome, micro-RNA, metabolome, and microbiome. In the last few years, researchers have developed new technologies and validated a wide range of salivary biomarkers that will soon make the use of saliva a clinical reality. However, a great need still exists for convenient and accurate point-of-care devices that can serve as a non-invasive diagnostic tool. In addition, there is an urgent need to decipher the scientific rationale and mechanisms that convey systemic diseases to saliva. Another promising technology called liquid biopsy enables detection of circulating tumor cells (CTCs) and fragments of tumor DNA in saliva, thus enabling non-invasive early detection of various cancers. The newly developed technology—electric field-induced release and measurement (EFIRM) provides near perfect detection of actionable mutations in lung cancer patients. These recent advances widened the salivary diagnostic approach from the oral cavity to the whole physiological system, and thus point towards a promising future of salivary diagnostics for personalized individual medicine applications including clinical decisions and post-treatment outcome predictions. Impact statement The purpose of this mini-review is to make an update about the present and future applications of saliva as a diagnostic biofluid in many fields of science such

  10. Commercial saliva collections tools

    National Research Council Canada - National Science Library

    Slowey, Paul D

    2013-01-01

    Saliva has been used as a specimen for diagnostics purposes for many years, but it has only been in the last 10 years that a number of new tools have been developed that promise to greatly increase...

  11. Commercial saliva collections tools.

    Science.gov (United States)

    Slowey, Paul D

    2013-02-01

    Saliva has been used as a specimen for diagnostics purposes for many years, but it has only been in the last 10 years that a number of new tools have been developed that promise to greatly increase the use of oral specimens for broad-based diagnosis and potentially screening applications. This article focuses on tools that are commercially viable or can play a role in whole saliva collection and future testing for critical diseases.

  12. Sources of pre-analytical variations in yield of DNA extracted from blood samples: analysis of 50,000 DNA samples in EPIC.

    Directory of Open Access Journals (Sweden)

    Elodie Caboux

    Full Text Available The European Prospective Investigation into Cancer and nutrition (EPIC is a long-term, multi-centric prospective study in Europe investigating the relationships between cancer and nutrition. This study has served as a basis for a number of Genome-Wide Association Studies (GWAS and other types of genetic analyses. Over a period of 5 years, 52,256 EPIC DNA samples have been extracted using an automated DNA extraction platform. Here we have evaluated the pre-analytical factors affecting DNA yield, including anthropometric, epidemiological and technical factors such as center of subject recruitment, age, gender, body-mass index, disease case or control status, tobacco consumption, number of aliquots of buffy coat used for DNA extraction, extraction machine or procedure, DNA quantification method, degree of haemolysis and variations in the timing of sample processing. We show that the largest significant variations in DNA yield were observed with degree of haemolysis and with center of subject recruitment. Age, gender, body-mass index, cancer case or control status and tobacco consumption also significantly impacted DNA yield. Feedback from laboratories which have analyzed DNA with different SNP genotyping technologies demonstrate that the vast majority of samples (approximately 88% performed adequately in different types of assays. To our knowledge this study is the largest to date to evaluate the sources of pre-analytical variations in DNA extracted from peripheral leucocytes. The results provide a strong evidence-based rationale for standardized recommendations on blood collection and processing protocols for large-scale genetic studies.

  13. Secretory proteins in the saliva of children.

    Science.gov (United States)

    Sivakumar, Thiruvanamalai; Hand, Arthur R; Mednieks, Maija

    2009-12-01

    The protein composition of oral fluid is modulated by environmental factors and physiological states, i.e. chemical, mechanical and pharmacologic stimuli, pathologic conditions, and psychological stress. Secretory protein concentrations in samples of whole saliva (WS) from children were measured and the results were subjected to statistical analysis. Protein expression was determined using electrophoresis and Western blotting. Protein profiles of children were significantly different from those of adults (n = 50, P saliva from children contained a group of high-molecular-weight (>90 kDa) proteins, whereas fewer than 5% of samples from adults had comparable bands. The ratio of the regulatory subunits (RII) of type II protein kinase A (an enzyme that regulates secretion) to total protein was stable in children's saliva, but variable in saliva from adults. Alpha amylase (alpha-amylase), an enzyme that digests carbohydrates, was less degraded in WS of children than in that of adults. Gingival crevicular fluid of both children and adults did not contain alpha-amylase or RII. No significant gender-based differences were found, but Caucasian children had higher salivary protein levels than children with an African background. Saliva collection is rapid, painless, non-invasive, economical, and yields findings that are reproducible. Objective, biochemical monitoring of secretory proteins in oral fluid of children may reveal responses to stressful stimuli.

  14. A cDNA Clone-Launched Platform for High-Yield Production of Inactivated Zika Vaccine

    Directory of Open Access Journals (Sweden)

    Yujiao Yang

    2017-03-01

    Full Text Available A purified inactivated vaccine (PIV using the Zika virus (ZIKV Puerto Rico strain PRVABC59 showed efficacy in monkeys, and is currently in a phase I clinical trial. High-yield manufacture of this PIV is essential for its development and vaccine access. Here we report an infectious cDNA clone-launched platform to maximize its yield. A single NS1 protein substitution (K265E was identified to increase ZIKV replication on Vero cells (a cell line approved for vaccine production for both Cambodian FSS13025 and Puerto Rico PRVABC59 strains. The NS1 mutation did not affect viral RNA synthesis, but significantly increased virion assembly through an increased interaction between NS1 and NS2A (a known regulator of flavivirus assembly. The NS1 mutant virus retained wild-type virulence in the A129 mouse model, but decreased its competence to infect Aedes aegypti mosquitoes. To further increase virus yield, we constructed an infectious cDNA clone of the clinical trial PIV strain PRVABC59 containing three viral replication-enhancing mutations (NS1 K265E, prM H83R, and NS3 S356F. The mutant cDNA clone produced >25-fold more ZIKV than the wild-type parent on Vero cells. This cDNA clone-launched manufacture platform has the advantages of higher virus yield, shortened manufacture time, and minimized chance of contamination.

  15. A cDNA Clone-Launched Platform for High-Yield Production of Inactivated Zika Vaccine.

    Science.gov (United States)

    Yang, Yujiao; Shan, Chao; Zou, Jing; Muruato, Antonio E; Bruno, Diniz Nunes; de Almeida Medeiros Daniele, Barbosa; Vasconcelos, Pedro F C; Rossi, Shannan L; Weaver, Scott C; Xie, Xuping; Shi, Pei-Yong

    2017-03-01

    A purified inactivated vaccine (PIV) using the Zika virus (ZIKV) Puerto Rico strain PRVABC59 showed efficacy in monkeys, and is currently in a phase I clinical trial. High-yield manufacture of this PIV is essential for its development and vaccine access. Here we report an infectious cDNA clone-launched platform to maximize its yield. A single NS1 protein substitution (K265E) was identified to increase ZIKV replication on Vero cells (a cell line approved for vaccine production) for both Cambodian FSS13025 and Puerto Rico PRVABC59 strains. The NS1 mutation did not affect viral RNA synthesis, but significantly increased virion assembly through an increased interaction between NS1 and NS2A (a known regulator of flavivirus assembly). The NS1 mutant virus retained wild-type virulence in the A129 mouse model, but decreased its competence to infect Aedes aegypti mosquitoes. To further increase virus yield, we constructed an infectious cDNA clone of the clinical trial PIV strain PRVABC59 containing three viral replication-enhancing mutations (NS1 K265E, prM H83R, and NS3 S356F). The mutant cDNA clone produced >25-fold more ZIKV than the wild-type parent on Vero cells. This cDNA clone-launched manufacture platform has the advantages of higher virus yield, shortened manufacture time, and minimized chance of contamination. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  16. 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...... there were no cultivable bacteria, whereas bacteria were cultivated in all positive control samples. In eight of 10 PCR samples no bacterial DNA was detected. The most frequent bacteria in the remaining non-sterile parotid saliva samples and positive control samples were non-haemolytical streptococci...

  17. Predictors of mother and child DNA yields in buccal cell samples collected in pediatric cancer epidemiologic studies: a report from the Children’s Oncology group

    Science.gov (United States)

    2013-01-01

    Background Collection of high-quality DNA is essential for molecular epidemiology studies. Methods have been evaluated for optimal DNA collection in studies of adults; however, DNA collection in young children poses additional challenges. Here, we have evaluated predictors of DNA quantity in buccal cells collected for population-based studies of infant leukemia (N = 489 mothers and 392 children) and hepatoblastoma (HB; N = 446 mothers and 412 children) conducted through the Children’s Oncology Group. DNA samples were collected by mail using mouthwash (for mothers and some children) and buccal brush (for children) collection kits and quantified using quantitative real-time PCR. Multivariable linear regression models were used to identify predictors of DNA yield. Results Median DNA yield was higher for mothers in both studies compared with their children (14 μg vs. <1 μg). Significant predictors of DNA yield in children included case–control status (β = −0.69, 50% reduction, P = 0.01 for case vs. control children), brush collection type, and season of sample collection. Demographic factors were not strong predictors of DNA yield in mothers or children in this analysis. Conclusions The association with seasonality suggests that conditions during transport may influence DNA yield. The low yields observed in most children in these studies highlight the importance of developing alternative methods for DNA collection in younger age groups. PMID:23937514

  18. High Precision and High Yield Fabrication of Dense Nanoparticle Arrays onto DNA Origami at Statistically Independent Binding Sites †

    Science.gov (United States)

    Takabayashi, Sadao; Klein, William P.; Onodera, Craig; Rapp, Blake; Flores-Estrada, Juan; Lindau, Elias; Snowball, Lejmarc; Sam, Joseph Tyler; Padilla, Jennifer E.; Lee, Jeunghoon; Knowlton, William B.; Graugnard, Elton; Yurke, Bernard; Kuang, Wan; Hughes, William L.

    2015-01-01

    High precision, high yield, and high density self-assembly of nanoparticles into arrays is essential for nanophotonics. Spatial deviations as small as a few nanometers can alter the properties of near-field coupled optical nanostructures. Several studies have reported assemblies of few nanoparticle structures with controlled spacing using DNA nanostructures with variable yield. Here, we report multi-tether design strategies and attachment yields for homo- and hetero-nanoparticle arrays templated by DNA origami nanotubes. Nanoparticle attachment yield via DNA hybridization is comparable with streptavidin-biotin binding. Independent of the number of binding sites, >97% site-occupation was achieved with four tethers and 99.2% site-occupation is theoretically possible with five tethers. The interparticle distance was within 2 nm of all design specifications and the nanoparticle spatial deviations decreased with interparticle spacing. Modified geometric, binomial, and trinomial distributions indicate that site-bridging, steric hindrance, and electrostatic repulsion were not dominant barriers to self-assembly and both tethers and binding sites were statistically independent at high particle densities. PMID:25311051

  19. Saliva: a fluid of study for OMICS.

    Science.gov (United States)

    Cuevas-Córdoba, Betzaida; Santiago-García, Juan

    2014-02-01

    Saliva is a fluid that can be collected easily and noninvasively. Its functions in the oral cavity are well known. Advances in molecular biology and technology, as well as research conducted by the various disciplines of omics (genomics, transcriptomics, proteomics, metabolomics, and metagenomics) have contributed to the identification and characterization of salivary components, including DNA, RNA, proteins, metabolites, and microorganisms. These biomolecules enter the saliva through extracellular and intracellular routes, providing information from several organs and systems and raising the possibility of their use as disease biomarkers. In recent years, these factors have expanded the potential use of saliva as a diagnostic fluid for oral and systemic diseases. This review integrates information regarding salivary biomolecules studied through omics and explores their utility as biomarkers for the diagnosis of several infectious and noninfectious diseases, and the opportunity they represent for the development of point of care devices for clinical application. We also discuss the advantages, disadvantages, and challenges to be overcome in order to establish saliva as a useful fluid for the accurate diagnosis and monitoring of a wide range of diseases.

  20. Extracting DNA from 'jaws': High yield and quality from archived tiger shark (Galeocerdo cuvier) skeletal material

    DEFF Research Database (Denmark)

    Eg Nielsen, Einar; Morgan, J. A T; Maher, S. L.

    2017-01-01

    Archived specimens are highly valuable sources of DNA for retrospective genetic/genomic analysis. However, often limited effort has been made to evaluate and optimize extraction methods, which may be crucial for downstream applications. Here, we assessed and optimized the usefulness of abundant...... archived skeletal material from sharks as a source of DNA for temporal genomic studies. Six different methods for DNA extraction, encompassing two different commercial kits and three different protocols, were applied to material, so-called bio-swarf, from contemporary and archived jaws and vertebrae...

  1. 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 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.

  2. Burning mouth and saliva.

    Science.gov (United States)

    Chimenos-Kustner, Eduardo; Marques-Soares, Maria Sueli

    2002-01-01

    Stomatodynia is the complaint of burning, tickling or itching of the oral cavity, and can be associated with other oral and non-oral signs and symptoms. However, the oral mucosa often appears normal, with no apparent underlying organic cause to account for the symptomatology. The etiology is unknown, though evidence points to the participation of numerous local, systemic and psychological factors. Among the local factors, saliva may play an important role in the symptoms of burning mouth. Saliva possesses specific rheological properties as a result of its chemical, physical and biological characteristics - these properties being essential for maintaining balanced conditions within the oral cavity. Patients with burning mouth present evidence of changes in salivary composition and flow, as well as a probable alteration in the oral mucosal sensory perception related particularly to dry mouth and taste alterations. On the other hand, alterations in salivary composition appear to reflect on its viscosity and symptomatology of burning mouth. Saliva is a field open to much research related to burning mouth, and knowledge of its properties (e.g., viscosity) merits special attention in view of its apparent relationship to the symptoms of burning mouth. The present study describes our clinical experience with burning mouth, and discusses some of the aspects pointing to salivary alterations as one of the most important factors underlying stomatodynia.

  3. The levels of yield and purity of genomic DNA from five tomato ...

    African Journals Online (AJOL)

    Isolation of good quality genomic DNA from different plant materials is an important prerequisite for many molecular techniques related to both basic and applied research in the areas of plant molecular biology, crop improvement, biodiversity studies and conservation of genetic materials. Therefore, the need to extract ...

  4. Bioinformatics advances in saliva diagnostics.

    Science.gov (United States)

    Ai, Ji-Ye; Smith, Barry; Wong, David T W

    2012-06-01

    There is a need recognized by the National Institute of Dental & Craniofacial Research and the National Cancer Institute to advance basic, translational and clinical saliva research. The goal of the Salivaomics Knowledge Base (SKB) is to create a data management system and web resource constructed to support human salivaomics research. To maximize the utility of the SKB for retrieval,integration and analysis of data, we have developed the Saliva Ontology and SDxMart. This article reviews the informatics advances in saliva diagnostics made possible by the Saliva Ontology and SDxMart.

  5. Boca ardiente y saliva

    OpenAIRE

    Chimenos Küstner, Eduardo; Marques Soares, Maria Sueli

    2002-01-01

    La sensación de ardor, escozor o picor generalizados en la cavidad bucal se denomina estomatodinia. Es un síntoma que puede guardar relación con otros síntomas o signos orales y no orales. Sin embargo, es frecuente que la mucosa bucal esté normal, no observándose una causa orgánica que justifique la sintomatología. Su etiología es desconocida, a pesar de que hay indicios de la participación de numerosos factores locales, sistémicos y psicológicos. Entre los factores locales, la saliva puede d...

  6. Effect of DNA extraction procedure, repeated extraction and ethidium monoazide (EMA)/propidium monoazide (PMA) treatment on overall DNA yield and impact on microbial fingerprints for bacteria, fungi and archaea in a reference soil.

    Science.gov (United States)

    Wagner, Andreas O; Praeg, Nadine; Reitschuler, Christoph; Illmer, Paul

    2015-09-01

    Different DNA extraction protocols were evaluated on a reference soil. A wide difference was found in the total extractable DNA as derived from different extraction protocols. Concerning the DNA yield phenol-chloroform-isomyl alcohol extraction resulted in high DNA yield but also in a remarkable co-extraction of contaminants making PCR from undiluted DNA extracts impossible. By comparison of two different extraction kits, the Macherey&Nagel SoilExtract II kit resulted in the highest DNA yields when buffer SL1 and the enhancer solution were applied. The enhancer solution not only significantly increased the DNA yield but also the amount of co-extracted contaminates, whereas additional disintegration strategies did not. Although a three times repeated DNA extraction increased the total amount of extracted DNA, microbial fingerprints were merely affected. However, with the 5th extraction this changed. A reduction of total DGGE band numbers was observed for archaea and fungi, whereas for bacteria the diversity increased. The application of ethidium monoazide (EMA) or propidium monoazide (PMA) treatment aiming on the selective removal of soil DNA derived from cells lacking cell wall integrity resulted in a significant reduction of total extracted DNA, however, the hypothesized effect on microbial fingerprints failed to appear indicating the need for further investigations.

  7. Predictors of mother and child DNA yields in buccal cell samples collected in pediatric cancer epidemiologic studies: a report from the Children's Oncology group.

    Science.gov (United States)

    Poynter, Jenny N; Ross, Julie A; Hooten, Anthony J; Langer, Erica; Blommer, Crystal; Spector, Logan G

    2013-08-12

    Collection of high-quality DNA is essential for molecular epidemiology studies. Methods have been evaluated for optimal DNA collection in studies of adults; however, DNA collection in young children poses additional challenges. Here, we have evaluated predictors of DNA quantity in buccal cells collected for population-based studies of infant leukemia (N = 489 mothers and 392 children) and hepatoblastoma (HB; N = 446 mothers and 412 children) conducted through the Children's Oncology Group. DNA samples were collected by mail using mouthwash (for mothers and some children) and buccal brush (for children) collection kits and quantified using quantitative real-time PCR. Multivariable linear regression models were used to identify predictors of DNA yield. Median DNA yield was higher for mothers in both studies compared with their children (14 μg vs. mothers or children in this analysis. The association with seasonality suggests that conditions during transport may influence DNA yield. The low yields observed in most children in these studies highlight the importance of developing alternative methods for DNA collection in younger age groups.

  8. Overexpression of sweetpotato expansin cDNA (IbEXP1) increases seed yield in Arabidopsis.

    Science.gov (United States)

    Bae, Jung Myung; Kwak, Man Sup; Noh, Seol Ah; Oh, Mi-Joung; Kim, Youn-Sung; Shin, Jeong Sheop

    2014-08-01

    Results of transcriptome analyses suggest that expansin genes play an active role in seed development and yield, but gain- or loss-of-function studies have not yet elucidated the functional role(s) of the expansin gene(s) in these processes. We have overexpressed a sweetpotato expansin gene (IbEXP1) in Arabidopsis under the control of cauliflower mosaic 35S promoter in an attempt to determine the effect of the expansin gene in seed development and yield in heterologous plants. The growth rate was enhanced in IbEXP1-overexpressing (ox) plants relative to wild-type Col-0 plants during early vegetative growth stage. At the reproductive stage, the number of rosette leaves was higher in IbEXP1-ox plants than that in Col-0 plants, and siliques were thicker. IbEXP1-ox plants produced larger seeds, accumulated more protein and starch in each seed, and produced more inflorescence stems and siliques than Col-0 plants, leading to a 2.1-2.5-fold increase in total seed yield per plant. The transcript level of IbEXP1 was up-regulated in response to brassinosteroid (BR) treatment in sweetpotato, and the transcript levels of three BR-responsive genes, fatty acid elongase 3-ketoacyl-CoA synthase 1, HAIKU1 and MINISEED3, were also increased in IbEXP1-ox Arabidopsis plants, suggesting a possible involvement of IbEXP1 in at least one of the BR signaling pathways. Based on these results, we suggest that overexpression of IbEXP1 gene in heterologous plants is effective in increasing seed size and number and, consequently, seed yield.

  9. Algorithm for post-clustering curation of DNA amplicon data yields reliable biodiversity estimates

    DEFF Research Database (Denmark)

    Froslev, Tobias Guldberg; Kjoller, Rasmus; Bruun, Hans Henrik

    2017-01-01

    DNA metabarcoding is promising for cost-effective biodiversity monitoring, but reliable diversity estimates are difficult to achieve and validate. Here we present and validate a method, called LULU, for removing erroneous molecular operational taxonomic units (OTUs) from community data derived...... soil from 130 sites in Denmark spanning major environmental gradients. OTU tables are produced with several different OTU definition algorithms and subsequently curated with LULU, and validated against field survey data. LULU curation consistently improves α-diversity estimates and other biodiversity...... metrics, and does not require a sequence reference database; thus, it represents a promising method for reliable biodiversity estimation....

  10. Radioimmunoassay of thyroxine in saliva

    Energy Technology Data Exchange (ETDEWEB)

    Putz, Z.; Vanuga, A.; Veleminsky, J. (Institute of Clinical Endocrinology, Lubochna (Czechoslovakia))

    1985-04-01

    A simple radioimmunoassay (RIA) for thyroxine (T/sub 4/) in saliva has been described. Fifty euthyroid control subjects, 14 euthyroid pregnant women, 23 thyreotoxic and 10 hypothyroid patients were examined. Serum T/sub 3/, T/sub 4/, thyroxine binding globulin (TBG) and TSH were measured simultaneously. The mean level of T/sub 4/ in saliva in controls was 1.10 +- 0.07 nmol/l. There was a good correlation between the saliva and serum T/sub 4/ concentrations (r = 0.74) and between saliva T/sub 4/ values and the T/sub 4//TBG ratio (r = 0.83). The saliva T/sub 4/ levels, like serum free T/sub 4/, were not dependent on fluctuations of serum TBG concentrations. In euthyroid pregnant women, saliva T/sub 4/ levels were within the normal range while the serum T/sub 4/ and TBG were increased. There was a good agreement of saliva T/sub 4/ values with the functional state of the thyroid. Thus, the RIA of saliva T/sub 4/ could replace the laborious determination of serum free T/sub 4/. It can especially be useful in instances with abnormal values of TBG, as it is in pregnancy, in congenital deficiency of serum TBG or in subjects with hereditary elevated TBG levels.

  11. 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....

  12. Saliva as a diagnostic fluid.

    Science.gov (United States)

    Samaranayake, Lakshman

    2007-10-01

    The use of saliva as a diagnostic fluid for various human ailments is gaining popularity as it offers distinct advantages over serum. These include the non-invasive nature of saliva collection compared with phlebotomy, simplicity of collection even for individuals with a modest training and the cost-effective applicability for screening large populations. Whole saliva is most frequently used for diagnosis of systemic diseases since it is readily collected and contains serum constituents while gland-specific saliva is useful for investigating pathology of major salivary glands. Broadly, saliva analysis is currently used for the diagnosis of infectious and malignant diseases, hereditary disorders, autoimmune diseases, and endocrine disorders, as well as for the assessment of therapeutic drug levels, particularly in monitoring drug abuse. This review addresses the current status of salivary diagnostics and their future potential.

  13. Tales from scales: old DNA yields insights into contemporary evolutionary processes affecting fishes.

    Science.gov (United States)

    Quinn, Thomas P; Seamons, Todd R

    2009-06-01

    Salmon and trout populations are suffering declines in abundance and diversity over much of their range around the Atlantic and Pacific rims as a consequence of many factors. One method of dealing with the decline has been to produce them in hatcheries but the wisdom of this approach has been hotly debated (e.g. Hilborn & Winton 1993; Waples 1999; Brannon et al. 2004). One concern is that domesticated hatchery strains will interbreed with locally adapted wild fish; but how do we study the genetic effects if the introgression might have occurred in the past? Hansen (2002) used DNA isolated from archived scales from brown trout, Salmo trutta (Fig. 1), to show that domesticated trout had, to varying degrees, genetically introgressed with wild, native trout in two Danish rivers. Extending that study, Hansen et al. (2009) have examined DNA from brown trout scales in six Danish rivers collected during historical (1927-1956) and contemporary (2000-2006) periods and from two hatchery source populations, to assess the effects of stocking nonlocal strains of hatchery trout and declining abundance on genetic diversity. Using 21 microsatellite loci, they revealed that genetic change occurred between the historic and contemporary time periods. Many populations appeared to have some low level of introgression from hatchery stocks and two populations apparently experienced high levels of introgression. Hansen et al. (2009) also showed that population structure persists in contemporary populations despite apparent admixture and migration among populations, providing evidence that the locally adapted populations have struggled against and, to some extent, resisted being overwhelmed by repeated introductions of and interbreeding with non-native, hatchery-produced conspecifics.

  14. Dried Saliva Spots: A Robust Method for Detecting Streptococcus pneumoniae Carriage by PCR

    Directory of Open Access Journals (Sweden)

    Cassandra L. Krone

    2016-03-01

    Full Text Available The earliest studies in the late 19th century on Streptococcus pneumoniae (S. pneumoniae carriage used saliva as the primary specimen. However, interest in saliva declined after the sensitive mouse inoculation method was replaced by conventional culture, which made isolation of pneumococci from the highly polymicrobial oral cavity virtually impossible. Here, we tested the feasibility of using dried saliva spots (DSS for studies on pneumococcal carriage. Saliva samples from children and pneumococcus-spiked saliva samples from healthy adults were applied to paper, dried, and stored, with and without desiccant, at temperatures ranging from −20 to 37 °C for up to 35 days. DNA extracted from DSS was tested with quantitative-PCR (qPCR specifically for S. pneumoniae. When processed immediately after drying, the quantity of pneumococcal DNA detected in spiked DSS from adults matched the levels in freshly spiked raw saliva. Furthermore, pneumococcal DNA was stable in DSS stored with desiccant for up to one month over a broad range of temperatures. There were no differences in the results when spiking saliva with varied pneumococcal strains. The collection of saliva can be a particularly useful in surveillance studies conducted in remote settings, as it does not require trained personnel, and DSS are resilient to various transportation conditions.

  15. DNA shuffling of adeno-associated virus yields functionally diverse viral progeny.

    Science.gov (United States)

    Koerber, James T; Jang, Jae-Hyung; Schaffer, David V

    2008-10-01

    Adeno-associated virus (AAV) vectors are extremely effective gene-delivery vehicles for a broad range of applications. However, the therapeutic efficacy of these and other vectors is currently limited by barriers to safe, efficient gene delivery, including pre-existing antiviral immunity, and infection of off-target cells. Recently, we have implemented directed evolution of AAV, involving the generation of randomly mutagenized viral libraries based on serotype 2 and high-throughput selection, to engineer enhanced viral vectors. Here, we significantly extend this capability by performing high-efficiency in vitro recombination to create a large (10(7)), diverse library of random chimeras of numerous parent AAV serotypes (AAV1, 2, 4-6, 8, and 9). In order to analyze the extent to which such highly chimeric viruses can be viable, we selected the library for efficient viral packaging and infection, and successfully recovered numerous novel chimeras. These new viruses exhibited a broad range of cell tropism both in vitro and in vivo and enhanced resistance to human intravenous immunoglobulin (IVIG), highlighting numerous functional differences between these chimeras and their parent serotypes. Thus, directed evolution can potentially yield unlimited numbers of new AAV variants with novel gene-delivery properties, and subsequent analysis of these variants can further extend basic knowledge of AAV biology.

  16. The functions of human saliva

    DEFF Research Database (Denmark)

    Dawes, C; Pedersen, Anne Marie Lynge; Villa, A

    2015-01-01

    as a buffer to protect oral, pharyngeal and oesophageal mucosae from orally ingested acid or acid regurgitated from the stomach. Saliva protects the teeth against acid by contributing to the acquired enamel pellicle, which forms a renewable lubricant between opposing tooth surfaces, by being supersaturated...... with respect to tooth mineral, by containing bicarbonate as a buffer and urea and by facilitating clearance of acidic materials from the mouth. Saliva contains many antibacterial, antiviral and antifungal agents which modulate the oral microbial flora in different ways. Saliva also facilitates the healing...

  17. [The diagnostic possibilities of saliva].

    Science.gov (United States)

    Kochurova, E V; Kozlov, S V

    2014-01-01

    Saliva is a clinically informative biological fluid which contains multitude of bio-markers. This characteristic makes it possible to carry out numerous analyzes for developing mode to test patient in situ, express-tests included. The diagnostic by saliva is a new area of more simple application both markers and analyzers that can be useful in diagnostic of diseases of oral cavity, oncological diseases included. The using of saliva expands perspectives for making clinical diagnosis and establishment of dynamics and monitoring of disease.

  18. Saliva fractions from South African Russian wheat aphid biotypes ...

    African Journals Online (AJOL)

    Abstract. The Russian wheat aphid (RWA), Diuraphis noxia (Kurdjomov, 1913), is a notorious pest that reduces yield in wheat. Nevertheless, the source of eliciting activity during RWA–wheat interaction has not been established. This paper reports on the isolation of eliciting activity in aphid saliva that is capable of inducing ...

  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. Real-time PCR quantification of six periodontal pathogens in saliva samples from healthy young adults.

    Science.gov (United States)

    Zhou, Xiaodong; Liu, Xiaoli; Li, Jing; Aprecio, Raydolfo M; Zhang, Wu; Li, Yiming

    2015-05-01

    The use of saliva as a diagnostic fluid for the evaluation of periodontal health has gained attention recently. Most published real-time PCR assays focused on quantification of bacteria in subgingival plaque, not in saliva. The aims of this study were to develop a real-time PCR assay for quantification of six periodontal pathogens in saliva and to establish a relationship between the amount of DNA (fg) and colony-forming unit (CFU). TaqMan primers/probe sets were used for the detection of Aggregatibacter actinomycetemcomitans (Aa), Eikenella corrodens (Ec), Fusobacterium nucleatum (Fn), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Tannerella forsythia (Tf), and total bacteria. Six periodontal pathogens and total bacteria in saliva from 24 periodontally healthy individuals were determined. The relationship between the amount of DNA (fg) and CFU was established by measuring the concentrations of extracted bacterial DNA and CFU per milliliter of bacteria on agar plates. Fn, Ec, and Pi were detected in all saliva samples, while 58.5, 45.8, and 33.3% were detected for Tf, Pg, and Aa, respectively. Numbers of Ec and Fn in saliva were highly correlated (R(2) = 0.93, P saliva and estimate the number of live bacteria (CFU). This real-time PCR assay in combination with the relationship between DNA (fg) and CFU has the potential to be an adjunct in evaluation of periodontal health status.

  1. Saliva as a diagnostic medium.

    Science.gov (United States)

    Pink, Richard; Simek, Jiri; Vondrakova, Jana; Faber, Edgar; Michl, Petr; Pazdera, Jindrich; Indrak, Karel

    2009-06-01

    This is a review of current knowledge on the use of saliva, gingival cervical fluid and mucosal transudate in the detection of some oral and systemic diseases as well as drugs. Oral fluid is a diagnostic medium that can be easily collected and with minimal invasion but it has been neglected in the past. Today, saliva is being used more often to diagnose: HIV virus, oro-facial and systemic tumors, cardiovascular disease and in detecting addictive substances. Neutropil levels in saliva may also indicate successful bone marrow transplant. Oral fluid is now systematically being researched and oral fluid analysis is being compared with the analysis of other diagnostic media such as blood and urine. A number of recent studies have focused on oncogenic marker detection and its monitoring in saliva. The latest clinical and laboratory findings on diagnostic markers of oropharyngeal carcinoma in oral fluid could be the beginning of their wider use as a diagnostic medium. Oral fluid can also be also used to diagnose other malignancies such as breast cancer which was one of the first malignant tumors to be detected using genetic protein biomarkers. Raised levels of CA15-3 and the epidermal growth factor (EGF) receptor have been found in patients with breast cancer and elevated levels of CA 125 and the glycoprotein complex in the saliva of ovarian cancer patients. Doubtless, the diagnostic value of saliva, aided by current technological development will increase rapidly in the near future.

  2. Saliva as a potential diagnostic tool

    OpenAIRE

    T Deepa; N Thirrunavukkarasu

    2010-01-01

    Saliva is a complex fluid consisting of secretions from the major and minor salivary glands. Gland-specific saliva can be used to diagnose any pathology from the specific major salivary gland. Whole saliva has serum constituents that are derived from the local vasculature of the salivary glands and gingival crevicular fluid. Saliva, as a diagnostic fluid, has distinctive advantages over serum as whole saliva can be collected non-invasively by individuals with limited training using simple equ...

  3. Evaluating differential nuclear DNA yield rates and osteocyte numbers among human bone tissue types: A synchrotron radiation micro-CT approach.

    Science.gov (United States)

    Andronowski, Janna M; Mundorff, Amy Z; Pratt, Isaac V; Davoren, Jon M; Cooper, David M L

    2017-05-01

    Molecular human identification has conventionally focused on DNA sampling from dense, weight-bearing cortical bone tissue, typically from femora or tibiae. A comparison of skeletal elements from three contemporary individuals demonstrated that elements with high quantities of cancellous bone yielded nuclear DNA at the highest rates, suggesting that preferentially sampling cortical bone may be suboptimal (Mundorff & Davoren, 2014). Despite these findings, the reason for the differential DNA yields between cortical and cancellous bone tissues remains unknown. The primary goal of this work is to ascertain whether differences in bone microstructure can be used to explain differential nuclear DNA yield among bone tissue types observed by Mundorff and Davoren (2014), with a focus on osteocytes and the three-dimensional (3D) quantification of their associated lacunae. Osteocytes and other bone cells are recognized to house DNA in bone tissue, thus examining the density of their lacunae may explain why nuclear DNA yield rates differ among bone tissue types. Lacunae were visualized and quantified using synchrotron radiation-based micro-Computed Tomographic imaging (SR micro-CT). Volumes of interest (VOIs) from cortical and cancellous bone tissues (n=129) were comparatively analyzed from the three skeletons sampled for Mundorff and Davoren's (2014) study. Analyses tested the primary hypothesis that the abundance and density of osteocytes (inferred from their lacunar spaces) vary between cortical and cancellous bone tissue types. Results demonstrated that osteocyte lacunar abundance and density vary between cortical and cancellous bone tissue types, with cortical bone VOIs containing a higher lacunar abundance and density. We found that the osteocyte lacunar density values are independent of nuclear DNA yield, suggesting an alternative explanation for the higher nuclear DNA yields from bones with greater quantities of cancellous bone tissue. The use of SR micro-CT allowed for

  4. The yield and quality of cellular and bacterial DNA extracts from human oral rinse samples are variably affected by the cell lysis methodology.

    Science.gov (United States)

    Sohrabi, Mohsen; Nair, Raj G; Samaranayake, Lakshman P; Zhang, Li; Zulfiker, Abu Hasanat Md; Ahmetagic, Adnan; Good, David; Wei, Ming Q

    2016-03-01

    Recent culture-independent studies have enabled detailed mapping of human microbiome that has not been hitherto achievable by culture-based methods. DNA extraction is a key element of bacterial culture-independent studies that critically impacts on the outcome of the detected microbial profile. Despite the variations in DNA extraction methods described in the literature, no standardized technique is available for the purpose of microbiome profiling. Hence, standardization of DNA extraction methods is urgently needed to yield comparable data from different studies. We examined the effect of eight different cell lysis protocols on the yield and quality of the extracted DNA from oral rinse samples. These samples were exposed to cell lysis techniques based on enzymatic, mechanical, and a combination of enzymatic-mechanical methods. The outcome measures evaluated were total bacterial population, Firmicutes levels and human DNA contamination (in terms of surrogate GAPDH levels). We noted that all three parameters were significantly affected by the method of cell lysis employed. Although the highest yield of gDNA was obtained using lysozyme-achromopeptidase method, the lysozyme-zirconium beads method yielded the peak quantity of total bacterial DNA and Firmicutes with a lower degree of GAPDH contamination compared with the other methods. Taken together our data clearly points to an urgent need for a consensus, standardized DNA extraction technique to evaluate the oral microbiome using oral rinse samples. Further, if Firmicutes levels are the focus of investigation in oral rinse microbiome analyses then the lysozyme-zirconium bead method would be the method of choice in preference to others. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Improved Yield of High Molecular Weight DNA Coincides with Increased Microbial Diversity Access from Iron Oxide Cemented Sub-Surface Clay Environments

    Science.gov (United States)

    Hurt, Richard A.; Robeson, Michael S.; Shakya, Migun; Moberly, James G.; Vishnivetskaya, Tatiana A.; Gu, Baohua; Elias, Dwayne A.

    2014-01-01

    Despite over three decades of progress, extraction of high molecular weight (HMW) DNA from high clay soils or iron oxide cemented clay has remained challenging. HMW DNA is desirable for next generation sequencing as it yields the most comprehensive coverage. Several DNA extraction procedures were compared from samples that exhibit strong nucleic acid adsorption. pH manipulation or use of alternative ion solutions offered no improvement in nucleic acid recovery. Lysis by liquid N2 grinding in concentrated guanidine followed by concentrated sodium phosphate extraction supported HMW DNA recovery from clays high in iron oxides. DNA recovered using 1 M sodium phosphate buffer (PB) as a competitive desorptive wash was 15.22±2.33 µg DNA/g clay, with most DNA consisting of >20 Kb fragments, compared to 2.46±0.25 µg DNA/g clay with the Powerlyzer system (MoBio). Increasing PB concentration in the lysis reagent coincided with increasing DNA fragment length during initial extraction. Rarefaction plots of 16S rRNA (V1–V3 region) pyrosequencing from A-horizon and clay soils showed an ∼80% and ∼400% larger accessed diversity compared to the Powerlyzer soil DNA system, respectively. The observed diversity from the Firmicutes showed the strongest increase with >3-fold more operational taxonomic units (OTU) recovered. PMID:25033199

  6. Improved yield of high molecular weight DNA coincides with increased microbial diversity access from iron oxide cemented sub-surface clay environments.

    Directory of Open Access Journals (Sweden)

    Richard A Hurt

    Full Text Available Despite over three decades of progress, extraction of high molecular weight (HMW DNA from high clay soils or iron oxide cemented clay has remained challenging. HMW DNA is desirable for next generation sequencing as it yields the most comprehensive coverage. Several DNA extraction procedures were compared from samples that exhibit strong nucleic acid adsorption. pH manipulation or use of alternative ion solutions offered no improvement in nucleic acid recovery. Lysis by liquid N2 grinding in concentrated guanidine followed by concentrated sodium phosphate extraction supported HMW DNA recovery from clays high in iron oxides. DNA recovered using 1 M sodium phosphate buffer (PB as a competitive desorptive wash was 15.22±2.33 µg DNA/g clay, with most DNA consisting of >20 Kb fragments, compared to 2.46±0.25 µg DNA/g clay with the Powerlyzer system (MoBio. Increasing PB concentration in the lysis reagent coincided with increasing DNA fragment length during initial extraction. Rarefaction plots of 16S rRNA (V1-V3 region pyrosequencing from A-horizon and clay soils showed an ∼80% and ∼400% larger accessed diversity compared to the Powerlyzer soil DNA system, respectively. The observed diversity from the Firmicutes showed the strongest increase with >3-fold more operational taxonomic units (OTU recovered.

  7. Streptococcus pneumoniae in saliva of Dutch primary school children.

    Science.gov (United States)

    Wyllie, Anne L; Chu, Mei Ling J N; Schellens, Mariëlle H B; van Engelsdorp Gastelaars, Jody; Jansen, Marc D; van der Ende, Arie; Bogaert, Debby; Sanders, Elisabeth A M; Trzciński, Krzysztof

    2014-01-01

    While nasopharyngeal sampling is the gold standard for the detection of Streptococcus pneumoniae carriage, historically seen, saliva sampling also seems highly sensitive for pneumococcal detection. We investigated S. pneumoniae carriage in saliva from fifty schoolchildren by conventional and molecular methods. Saliva was first culture-enriched for pneumococci, after which, DNA was extracted from all bacterial growth and tested by quantitative-PCR (qPCR) for pneumococcus-specific genes lytA and piaA. Next, serotype composition of the samples was determined by serotype-specific qPCRs, conventional-PCRs (cPCR) and sequencing of cPCR amplicons. Although only 2 (4%) of 50 samples were positive by conventional diagnostic culture, 44 (88%) were positive for pneumococci by qPCR. In total, we detected the presence of at least 81 pneumococcal strains representing 20 serotypes in samples from 44 carriers with 23 carriers (52%) positive for multiple (up to 6) serotypes. The number of serotypes detected per sample correlated with pneumococcal abundance. This study shows that saliva could be used as a tool for future pneumococcal surveillance studies. Furthermore, high rates of pneumococcal carriage and co-carriage of multiple pneumococcal strains together with a large number of serotypes in circulation suggests a ubiquitous presence of S. pneumoniae in saliva of school-aged children. Our results also suggest that factors promoting pneumococcal carriage within individual hosts may weaken competitive interactions between S. pneumoniae strains.

  8. Authentication of forensic DNA samples.

    Science.gov (United States)

    Frumkin, Dan; Wasserstrom, Adam; Davidson, Ariane; Grafit, Arnon

    2010-02-01

    Over the past twenty years, DNA analysis has revolutionized forensic science, and has become a dominant tool in law enforcement. Today, DNA evidence is key to the conviction or exoneration of suspects of various types of crime, from theft to rape and murder. However, the disturbing possibility that DNA evidence can be faked has been overlooked. It turns out that standard molecular biology techniques such as PCR, molecular cloning, and recently developed whole genome amplification (WGA), enable anyone with basic equipment and know-how to produce practically unlimited amounts of in vitro synthesized (artificial) DNA with any desired genetic profile. This artificial DNA can then be applied to surfaces of objects or incorporated into genuine human tissues and planted in crime scenes. Here we show that the current forensic procedure fails to distinguish between such samples of blood, saliva, and touched surfaces with artificial DNA, and corresponding samples with in vivo generated (natural) DNA. Furthermore, genotyping of both artificial and natural samples with Profiler Plus((R)) yielded full profiles with no anomalies. In order to effectively deal with this problem, we developed an authentication assay, which distinguishes between natural and artificial DNA based on methylation analysis of a set of genomic loci: in natural DNA, some loci are methylated and others are unmethylated, while in artificial DNA all loci are unmethylated. The assay was tested on natural and artificial samples of blood, saliva, and touched surfaces, with complete success. Adopting an authentication assay for casework samples as part of the forensic procedure is necessary for maintaining the high credibility of DNA evidence in the judiciary system.

  9. Increased EBV Shedding in Astronaut Saliva During Spaceflight

    Science.gov (United States)

    Pierson, D. L.; Stowe, R. P.; Phillips, T.; Lugg, D. J.; Mehta, S. K.

    2003-01-01

    Shedding of Epstein-Barr virus (EBV) by astronauts before, during, and after space shuttle missions was quantified. Of 1398 saliva specimens from 32 astronauts, 314 (23%) were positive for EBV DNA by PCR analysis. Of the saliva specimens collected before flight, 29% were positive for EBV DNA and of those collected during or after flight, 16% were EBV-positive. The number of EBV DNA copies from samples taken during the flight was 417+/-31, significantly higher (P EBV DNA with a frequency of 3.7% and a copy number of 40+/-2 per ml saliva. Ten days before flight and on landing day, antibody titers to EBV viral capsid antigen (VCA) were significantly (P < 0.05) higher than baseline levels. On landing day, urinary level of cortiso1 and catecholamines, and plasma levels of substance P and other neuropeptides, were increased over their preflight value. Results suggested that stress associated with spaceflight decreases cellular immunity and thereby leads to increased viral reactivation.

  10. [The acoustic indicator of saliva under stress].

    Science.gov (United States)

    Shalenkova, M A; Mikhaĭlova, Z D; Klemin, V A; Korkotashvili, L V; Abanin, A M; Klemina, A V; Dolgov, V V

    2014-03-01

    The situation of stress affects various organs and systems that results in development of functional disorders and/or somatic diseases. As a result, different noninvasive, including salivary, techniques of diagnostic of stress conditions are in the process of development. The dynamics of acoustic indicator of saliva is studied during the period of passing the exams. The relationship of indicator with levels of potassium, sodium, glucose and protein of saliva was analyzed. The sampling consisted of 102 students of 5 and 6 academic years of medical university. To detect the acoustic indicator of saliva acoustic analyzer AKBa-01- "BIOM" was applied. The level of potassium and sodium in saliva was detected using method of flame photometry. The level of glucose in saliva was detected by glucose oxydase technique using analyzer "EXAN-G". The protein in saliva was detected by biuretic technique. The correlation between acoustic indicator of saliva and analyzed indicators of saliva was established.

  11. 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.

  12. 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 ...

  13. MEASURING CHOLINESTERASE ACTIVITY IN HUMAN SALIVA.

    Science.gov (United States)

    To assess the potential for using saliva in pesticide biomonitoring, the consistency of cholinesterase activity in human saliva collected over time was examined. In this pilot study, saliva was collected from 20 healthy adults once per week for 5 consecutive weeks using 2 differe...

  14. 21 CFR 872.6050 - Saliva absorber.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Saliva absorber. 872.6050 Section 872.6050 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES DENTAL DEVICES Miscellaneous Devices § 872.6050 Saliva absorber. (a) Identification. A saliva...

  15. 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.

  16. Assessment of DNA contamination from dried blood spots and determination of DNA yield and function using archival newborn dried blood spots.

    Science.gov (United States)

    Cordovado, Suzanne K; Earley, Marie C; Hendrix, Miyono; Driscoll-Dunn, Rena; Glass, Michael; Mueller, Patricia W; Hannon, W Harry

    2009-04-01

    Residual dried blood spots (DBS) from newborn screening programs are often stored for years and are sometimes used for epidemiological studies. Because there is potential for DNA cross-contamination from specimen-to-specimen contact, we determined contamination levels following intentional contact and assessed archival DBS DNA degradation after storage in an uncontrolled environment. DBS from healthy adult females were rubbed with DBS from healthy or cystic fibrosis (CF)-affected adult males. Total human and male DNA was measured from the female DBS. Contamination levels were assessed using short tandem repeats (STRs). Female DBS contaminated with CF male DNA containing the F508del were analyzed for presence of this mutation. Archival DBS DNA amplification efficiency was determined using STR analysis. Most female DBS were contaminated, however only one specimen showed an incomplete STR profile consistent with contaminating CF-affected male DNA. Further testing by CF mutation screening was negative. DNA extracted from archival DBS showed robust amplification (range 100 bp-320 bp). Lightly abrasive contact between DBS resulted in DNA cross-contamination. The contaminating DNA did not interfere in CF-mutation tests; however this should be determined for individual assays. Since DNA from archival DBS robustly amplifies, newborn DBS could provide an invaluable resource for public health studies.

  17. A method for selectively enriching microbial DNA from contaminating vertebrate host DNA.

    Directory of Open Access Journals (Sweden)

    George R Feehery

    Full Text Available DNA samples derived from vertebrate skin, bodily cavities and body fluids contain both host and microbial DNA; the latter often present as a minor component. Consequently, DNA sequencing of a microbiome sample frequently yields reads originating from the microbe(s of interest, but with a vast excess of host genome-derived reads. In this study, we used a methyl-CpG binding domain (MBD to separate methylated host DNA from microbial DNA based on differences in CpG methylation density. MBD fused to the Fc region of a human antibody (MBD-Fc binds strongly to protein A paramagnetic beads, forming an effective one-step enrichment complex that was used to remove human or fish host DNA from bacterial and protistan DNA for subsequent sequencing and analysis. We report enrichment of DNA samples from human saliva, human blood, a mock malaria-infected blood sample and a black molly fish. When reads were mapped to reference genomes, sequence reads aligning to host genomes decreased 50-fold, while bacterial and Plasmodium DNA sequences reads increased 8-11.5-fold. The Shannon-Wiener diversity index was calculated for 149 bacterial species in saliva before and after enrichment. Unenriched saliva had an index of 4.72, while the enriched sample had an index of 4.80. The similarity of these indices demonstrates that bacterial species diversity and relative phylotype abundance remain conserved in enriched samples. Enrichment using the MBD-Fc method holds promise for targeted microbiome sequence analysis across a broad range of sample types.

  18. A novel method for detecting polymerase chain reaction product utilizing the 5 prime yields 3 prime exonuclease activity of Thermus aquaticus DNA polymerase

    Energy Technology Data Exchange (ETDEWEB)

    Holland, P.M.; Watson, R.; Abramson, R.D.; Gelfand, D.H. (Cetus Corp., Emeryville, CA (United States))

    1991-03-11

    The polymerase chain reaction (PCR) method of DNA amplification is a powerful and sensitive technique which has been greatly simplified by the use of the thermostable enzyme Thermus aquaticus (Taq) DNA polymerase. In addition to its polymerase activity, Taq DNA polymerase also has a 5{prime}{yields}3{prime} exonuclease activity. Utilizing this activity, the authors have developed a PCR detection method which generates signal simultaneously with target sequence amplification. No additional probing, blotting or hybridization assays of amplified product are necessary. A 5{prime}{yields}3{prime} exonuclease cleaves 5{prime} terminal nucleotides of nicked double stranded DNA. The authors have generated a substrate suitable for exonuclease activity in a PCR assay by the addition of a labeled oligonucleotide probe designed to hybridize within the target sequence. During amplification, the 5{prime}{yields}3{prime} exonuclease activity of Taq DNA polymerase degrades the probe into smaller fragments which can be differentiated from undegraded probe. The presence of probe does not influence PCR product formation. Hydrolysis occurs only when probe is bound specifically to template. The presence of high complexity DNA in the PCR mixture does not compromise the specificity of probe interaction. In a PCR assay, the amount of detectable label may be modified by altering cycle number, target copy number or probe concentration. The size of labeled fragments may also be modified by varying base composition of the probe. This detection method is advantageous over present day procedures in that it is neither labor intensive nor requires significant skills. The minimal sample handling could reduce the risk of sample contamination, thereby increasing accuracy.

  19. Saliva as a potential diagnostic tool.

    Science.gov (United States)

    Deepa, T; Thirrunavukkarasu, N

    2010-07-01

    Saliva is a complex fluid consisting of secretions from the major and minor salivary glands. Gland-specific saliva can be used to diagnose any pathology from the specific major salivary gland. Whole saliva has serum constituents that are derived from the local vasculature of the salivary glands and gingival crevicular fluid. Saliva, as a diagnostic fluid, has distinctive advantages over serum as whole saliva can be collected non-invasively by individuals with limited training using simple equipments. This review aimed to explore the diagnostic applications of saliva in systemic and oral diseases. Analysis of saliva can offer a cost-effective approach to screen for a larger population. Salivary analysis may be useful for diagnosing systemic oral disorders, as well as for monitoring hormone and therapeutic levels of drug.

  20. 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.

  1. SALIVA SEBAGAI UJI SARING OSTEOPOROSIS

    Directory of Open Access Journals (Sweden)

    Niniarty Z. Djamal

    2015-07-01

    Full Text Available Osteoporosis is a metabolic bone disease, and is characterized by low bone mass and microstructure deterioration of the bone, which leads to increased risk of fracture. Biomarker of bone metabolism can be seen as beginning of bone loss and first detection before imbalanced bone turnover comes. Biomarker of bone formation as serum bone alkaline fosfatase, osteocalcin (OC, procollagen type I, and biomarker of bone resorption as urine pyridinoline (Pyd and deoxypyridinoline (Dpd crosslinks, hydroxyprolin. The simultaneous examination of serum OC and urine Pyd or Dpd as a very good screening test for determination of bone imbalanced at the moment of the menopausal or the beginning of the pasca menopausal. Saliva as a potential diagnostic fluid for the assessment of osteoporosis biomarker concentrations. The study found elevated three classic warning signs for osteopororsis os OC, Dpd and 116 in the saliva of sheep without ovaries, which were similar to the levels of signs found in their blood and urine. Expectations, that the test may become available within five years and one day the test may be able to be performed at home like pregnancy test. Osteoporosis biomarker in saliva suggested detected of bone mass density easier. Beside that can be used as a method of early diagnostic and as a monitor therapy that as salinity of the examinations of bone mass on radiology.

  2. The impact of DNA contamination of bone samples in forensic case analysis and anthropological research.

    Science.gov (United States)

    von Wurmb-Schwark, Nicole; Heinrich, Anke; Freudenberg, Mechthild; Gebühr, Michael; Schwark, Thorsten

    2008-05-01

    Contamination precautions and quality control are great issues when human bones are investigated genetically. This is especially true for historical samples with only minute amounts of usually highly degraded DNA. But also in forensic routine analysis, sometimes DNA has to be isolated from bones in equally bad conditions, e.g. from burned victims. In such cases, there are several eventualities to contaminate the sample with foreign DNA, for example caused by the recovery of the bones, by trace investigation on a crime scene, or - of course - during handling in the lab. We present the investigation of artificially contaminated historical bone samples which contained no original DNA. Three different kind of contamination were studied: (1) touching of the samples, (2) application of saliva, and (3) application of pure DNA. The samples were genetically investigated without and with the employment of a defined cleaning protocol of the bones. The results show that pure DNA can usually not be removed from the bones and that saliva is a similar thread for subsequent DNA analysis. After the cleaning procedure about 70% of saliva contaminated samples still yielded reproducible STR profiles implicating severe problems for the investigation of highly degraded bone fragments. Simple touching of the specimens seems not to be a real problem for genetic investigations since the obtained signals were not reproducible.

  3. Development of Plant Expression Vector with Taq DNA Polymerase Gene to Yield Heat-tolerant Maize Lines

    National Research Council Canada - National Science Library

    Ya-Jun Zhang; Ling-Yan Zhou; Yu-Xian Bai; Wen-Bin Yi; Kun Cu; Xiaoxiong Nie; Xue-Lian Liang

    2017-01-01

    In this study, we have focused on using Taq DNA polymerase gene to develop heat-tolerant maize transgenic lines, which would aid in controlling crop loss due to heat stress and potentially increase the economic value...

  4. Saliva as a diagnostic fluid. Literature review

    OpenAIRE

    Martí-Álamo, Silvia; Mancheño-Franch, Aisha; Marzal-Gamarra, Cristina; Carlos-Fabuel, Laura

    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 amount of desqu...

  5. 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 ...

  6. The diagnostic role of saliva: a review

    OpenAIRE

    Mittal, Sanjeev; Bansal, Vikram; Garg, Shushant K.; Atreja, Gaurav; Bansal, Sanjay

    2011-01-01

    As a diagnostic fluid, saliva offers distinctive advantages over serum because it can be collected non-invasively by individuals, even by patient. Does not require special equipment for collection and storage as unlike blood saliva does not clot. Advantageous for person in whom blood drawing is difficult as in obese and haemophilic patient. Whole saliva used for diagnosis of systemic diseases, because it contains serum constituents. These constituents are derived from the ...

  7. Detection of Zika virus in saliva.

    Science.gov (United States)

    Musso, Didier; Roche, Claudine; Nhan, Tu-Xuan; Robin, Emilie; Teissier, Anita; Cao-Lormeau, Van-Mai

    2015-07-01

    During the largest Zika virus (ZIKV) outbreak ever reported that occurred from October 2013 to March 2014 in French Polynesia, we observed that several patients presenting the symptoms of acute phase Zika fever were tested negative in blood by ZIKV real-time PCR (RT-PCR). As we have previously detected ZIKV RNA in the saliva of a young child, we investigated the use of saliva as an alternative sample for routine ZIKV RNA detection. Over a 6 month period, 1,067 samples collected from 855 patients presenting symptoms of Zika fever (saliva only, blood only or both samples) were tested using a specific ZIKV RT-PCR. A medical questionnaire was available for most of the patients. ZIKV was more frequently detected in saliva compared to blood. For the 182 patients with both samples collected, tests were positive for 35 (19.2%) in saliva while negative in blood and tests were positive for 16 (8.8%) in blood while negative in saliva; the difference in mean days after symptoms onset and the percentage of the main symptoms of Zika fever for patients only positive in saliva or in blood was not significant. The use of saliva sample increased the rate of molecular detection of ZIKV at the acute phase of the disease but did not enlarge the window of detection of ZIKV RNA. Saliva was of particular interest when blood was difficult to collect (children and neonates especially). Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Heterologous expression of laccase cDNA from Ceriporiopsis subvermispora yields copper-activated apoprotein and complex isoform patterns

    Science.gov (United States)

    Luis F. Larrondo; Marcela Avila; Loreto Salas; Dan Cullen; Rafael Vicuna

    2003-01-01

    Analysis of genomic clones encoding a putative laccase in homokaryon strains of Ceriporiopsis subvermispora led to the identification of an allelic variant of the previously described lcs-1 gene. A cDNA clone corresponding to this gene was expressed in Aspergillus nidulans and in Aspergillus niger. Enzyme assays and Western blots showed that both hosts secreted active...

  9. HUBUNGAN FAKTOR RESIKO KARIES DALAM SALIVA DENGAN INDEKS DMF-T PADA PENDERITA DM TIPE II DI RSCM SUB BAGIAN ENDOKRIN (Laporan Penelitian)

    OpenAIRE

    Buddiwati Punta; Edi Hartini Sundoro

    2015-01-01

    Dental caries is a multifactorial disease. The most important factors in the development of caries is saliva. Reduced salivary secretion in patients with type 2 diabetes melitus to the occurrence of caries have yielded controversial results. The aim of the study was to find out whether the risk factors of the saliva i.e. saliva secretion rate, buffer capacity, salivary S. mutans, salivary Lactobacilli, alone or in combination, could be used for prediction of caries activity. Thirty patients w...

  10. Analysis of the shedding of three β-herpesviruses in urine and saliva of children with renal disease.

    Science.gov (United States)

    Yamamoto, Yasuto; Morooka, Masashi; Hashimoto, Shuji; Ihra, Masaru; Yoshikawa, Tetsushi

    2014-03-01

    Cytomegalovirus (CMV), human herpesvirus 6 (HHV-6) and 7 (HHV-7) are important pathogens in immunocompromised patients. To elucidate the kinetics of the three β-herpesviruses in saliva and urine samples were collected serially from children with renal diseases. Twenty children with renal diseases were enrolled in this study. A total of 240 saliva and urine samples were collected monthly from the patients over a 1-year period. Viral DNAs loads were measured by real-time PCR. In 10 CMV seropositive patients CMV DNA was detected rarely in saliva and CMV DNA load was lower than the other two β-herpesviruses DNA loads. All patients were seropositive for HHV-6B and the virus was detected frequently in saliva. Two of 20 patients were HHV-7 seronegative. High copies of viral DNA were detected continuously in saliva of the HHV-7 seropositive patients. Although neither CMV nor HHV-6B DNA load was different among the three renal diseases, HHV-7 DNA load was different among the diseases (P = 0.039). HHV-6B DNA loads were significantly higher in patients with immunosuppressive treatment compared to those without treatment (P = 0.013). Although CMV DNA was detected in urine samples collected from 5 of 10 CMV seropositive patients, HHV-6B and HHV-7 DNA were detected at relatively low frequencies in urine. No remarkable temporal associations between viral DNA excretion and proteinuria or immunosuppressive treatment were demonstrated. The pattern of viral DNA excretion in saliva and urine were different among the three viruses. No temporal correlation was observed between viral infection and renal diseases. © 2013 Wiley Periodicals, Inc.

  11. Initial yields of DNA double-strand breaks and DNA Fragmentation patterns depend on linear energy transfer in tobacco BY-2 protoplasts irradiated with helium, carbon and neon ions.

    Science.gov (United States)

    Yokota, Yuichiro; Yamada, Shinya; Hase, Yoshihiro; Shikazono, Naoya; Narumi, Issay; Tanaka, Atsushi; Inoue, Masayoshi

    2007-01-01

    The ability of ion beams to kill or mutate plant cells is known to depend on the linear energy transfer (LET) of the ions, although the mechanism of damage is poorly understood. In this study, DNA double-strand breaks (DSBs) were quantified by a DNA fragment-size analysis in tobacco protoplasts irradiated with high-LET ions. Tobacco BY-2 protoplasts, as a model of single plant cells, were irradiated with helium, carbon and neon ions having different LETs and with gamma rays. After irradiation, DNA fragments were separated into sizes between 1600 and 6.6 kbp by pulsed-field gel electrophoresis. Information on DNA fragmentation was obtained by staining the gels with SYBR Green I. Initial DSB yields were found to depend on LET, and the highest relative biological effectiveness (about 1.6) was obtained at 124 and 241 keV/microm carbon ions. High-LET carbon and neon ions induced short DNA fragments more efficiently than gamma rays. These results partially explain the large biological effects caused by high-LET ions in plants.

  12. Saliva Parameters and Erosive Wear in Adolescents

    NARCIS (Netherlands)

    Zwier, N.; Huysmans, M. C. D. N. J. M.; Jager, D. H. J.; Ruben, J.; Bronkhorst, E. M.; Truin, G. J.

    2013-01-01

    The aim of this study was to investigate the relationship between several parameters of saliva and erosive wear in adolescents. (Un-)stimulated saliva was collected from 88 adolescents with erosion and 49 controls (age 16 +/- 1 years). Flow rate, pH and buffer capacity were determined immediately.

  13. 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

  14. A potential method for non-invasive acute myocardial infarction detection based on saliva Raman spectroscopy and multivariate analysis

    Science.gov (United States)

    Cao, Gang; Chen, Maowen; Chen, Yuanxiang; Huang, Zufang; Lin, Jinyong; Lin, Jia; Xu, Zhihong; Wu, Shanshan; Huang, Wei; Weng, Guoxing; Chen, Guannan

    2015-12-01

    Raman spectroscopy (RS) was employed for human saliva biochemical analysis with the aim to develop a rapidly non-invasive test for acute myocardial infarction (AMI) detection. High-quality Raman spectra were obtained from human saliva samples of 46 AMI patients and 43 healthy controls. Significant differences in Raman intensities of prominent bands were observed between AMI and normal saliva. The tentative assignment of the observed Raman bands indicated constituent and conformational differences between the two groups. Furthermore, principal component analysis (PCA) combined with linear discriminant analysis (LDA) was employed to analyze and classify the Raman spectra acquired from AMI and healthy saliva, yielding a diagnostic sensitivity of 80.4% and specificity of 81.4%. The results from this exploratory study demonstrated the feasibility and potential for developing RS analysis of human saliva into a clinical tool for rapid AMI detection and screening.

  15. A New Method for Noninvasive Genetic Sampling of Saliva in Ecological Research.

    Directory of Open Access Journals (Sweden)

    Diana Lobo

    Full Text Available Noninvasive samples for genetic analyses have become essential to address ecological questions. Popular noninvasive samples such as faeces contain degraded DNA which may compromise genotyping success. Saliva is an excellent alternative DNA source but scarcity of suitable collection methods makes its use anecdotal in field ecological studies. We develop a noninvasive method of collection that combines baits and porous materials able to capture saliva. We report its potential in optimal conditions, using confined dogs and collecting saliva early after deposition. DNA concentration in saliva extracts was generally high (mean 14 ng μl(-1. We correctly identified individuals in 78% of samples conservatively using ten microsatellite loci, and 90% of samples using only eight loci. Consensus genotypes closely matched reference genotypes obtained from hair DNA (99% of identification successes and 91% of failures. Mean genotyping effort needed for identification using ten loci was 2.2 replicates. Genotyping errors occurred at a very low frequency (allelic dropout: 2.3%; false alleles: 1.5%. Individual identification success increased with duration of substrate handling inside dog's mouth and the volume of saliva collected. Low identification success was associated with baits rich in DNA-oxidant polyphenols and DNA concentrations <1 ng μl(-1. The procedure performed at least as well as other noninvasive methods, and could advantageously allow detection of socially low-ranked individuals underrepresented in sources of DNA that are involved in marking behaviour (faeces or urine. Once adapted and refined, there is promise for this technique to allow potentially high rates of individual identification in ecological field studies requiring noninvasive sampling of wild vertebrates.

  16. A New Method for Noninvasive Genetic Sampling of Saliva in Ecological Research

    Science.gov (United States)

    Lobo, Diana; Godinho, Raquel; Álvares, Francisco; López-Bao, José V.; Rodríguez, Alejandro

    2015-01-01

    Noninvasive samples for genetic analyses have become essential to address ecological questions. Popular noninvasive samples such as faeces contain degraded DNA which may compromise genotyping success. Saliva is an excellent alternative DNA source but scarcity of suitable collection methods makes its use anecdotal in field ecological studies. We develop a noninvasive method of collection that combines baits and porous materials able to capture saliva. We report its potential in optimal conditions, using confined dogs and collecting saliva early after deposition. DNA concentration in saliva extracts was generally high (mean 14 ng μl-1). We correctly identified individuals in 78% of samples conservatively using ten microsatellite loci, and 90% of samples using only eight loci. Consensus genotypes closely matched reference genotypes obtained from hair DNA (99% of identification successes and 91% of failures). Mean genotyping effort needed for identification using ten loci was 2.2 replicates. Genotyping errors occurred at a very low frequency (allelic dropout: 2.3%; false alleles: 1.5%). Individual identification success increased with duration of substrate handling inside dog’s mouth and the volume of saliva collected. Low identification success was associated with baits rich in DNA-oxidant polyphenols and DNA concentrations <1 ng μl-1. The procedure performed at least as well as other noninvasive methods, and could advantageously allow detection of socially low-ranked individuals underrepresented in sources of DNA that are involved in marking behaviour (faeces or urine). Once adapted and refined, there is promise for this technique to allow potentially high rates of individual identification in ecological field studies requiring noninvasive sampling of wild vertebrates. PMID:26496352

  17. Saliva in forensic odontology: A comprehensive update.

    Science.gov (United States)

    Saxena, Susmita; Kumar, Sanjeev

    2015-01-01

    In recent years, saliva has attracted much interest among researchers especially in the field of forensic sciences. This complex body fluid is gaining popularity due to its ease of collection, safety in handling and its close relationship with plasma. Analysis of saliva for serological testing and cellular content has proved to be of wide use in crime detection, drug and alcohol abuse, hormone identification, cases of poisoning and animal bites. There is a need for forensic laboratories to automate the settings specific for saliva as routinely done for blood or urine in order to consider saliva as the primary investigating tool in the absence of other body fluids. This update is aimed at highlighting the many uses of saliva in the practice of forensic odontology.

  18. The saliva microbiome of Pan and Homo.

    Science.gov (United States)

    Li, Jing; Nasidze, Ivan; Quinque, Dominique; Li, Mingkun; Horz, Hans-Peter; André, Claudine; Garriga, Rosa M; Halbwax, Michel; Fischer, Anne; Stoneking, Mark

    2013-09-11

    It is increasingly recognized that the bacteria that live in and on the human body (the microbiome) can play an important role in health and disease. The composition of the microbiome is potentially influenced by both internal factors (such as phylogeny and host physiology) and external factors (such as diet and local environment), and interspecific comparisons can aid in understanding the importance of these factors. To gain insights into the relative importance of these factors on saliva microbiome diversity, we here analyze the saliva microbiomes of chimpanzees (Pan troglodytes) and bonobos (Pan paniscus) from two sanctuaries in Africa, and from human workers at each sanctuary. The saliva microbiomes of the two Pan species are more similar to one another, and the saliva microbiomes of the two human groups are more similar to one another, than are the saliva microbiomes of human workers and apes from the same sanctuary. We also looked for the existence of a core microbiome and find no evidence for a taxon-based core saliva microbiome for Homo or Pan. In addition, we studied the saliva microbiome from apes from the Leipzig Zoo, and found an extraordinary diversity in the zoo ape saliva microbiomes that is not found in the saliva microbiomes of the sanctuary animals. The greater similarity of the saliva microbiomes of the two Pan species to one another, and of the two human groups to one another, are in accordance with both the phylogenetic relationships of the hosts as well as with host physiology. Moreover, the results from the zoo animals suggest that novel environments can have a large impact on the microbiome, and that microbiome analyses based on captive animals should be viewed with caution as they may not reflect the microbiome of animals in the wild.

  19. Right-handed double-helix ultrashort DNA yields chiral nematic phases with both right- and left-handed director twist

    Science.gov (United States)

    Zanchetta, Giuliano; Giavazzi, Fabio; Nakata, Michi; Buscaglia, Marco; Cerbino, Roberto; Clark, Noel A.; Bellini, Tommaso

    2010-01-01

    Concentrated solutions of duplex-forming DNA oligomers organize into various mesophases among which is the nematic (N∗), which exhibits a macroscopic chiral helical precession of molecular orientation because of the chirality of the DNA molecule. Using a quantitative analysis of the transmission spectra in polarized optical microscopy, we have determined the handedness and pitch of this chiral nematic helix for a large number of sequences ranging from 8 to 20 bases. The B-DNA molecule exhibits a right-handed molecular double-helix structure that, for long molecules, always yields N∗ phases with left-handed pitch in the μm range. We report here that ultrashort oligomeric duplexes show an extremely diverse behavior, with both left- and right-handed N∗ helices and pitches ranging from macroscopic down to 0.3 μm. The behavior depends on the length and the sequence of the oligomers, and on the nature of the end-to-end interactions between helices. In particular, the N∗ handedness strongly correlates with the oligomer length and concentration. Right-handed phases are found only for oligomers shorter than 14 base pairs, and for the sequences having the transition to the N∗ phase at concentration larger than 620 mg/mL. Our findings indicate that in short DNA, the intermolecular double-helical interactions switch the preferred liquid crystal handedness when the columns of stacked duplexes are forced at high concentrations to separations comparable to the DNA double-helix pitch, a regime still to be theoretically described. PMID:20876125

  20. Saliva stimulation with glycerine and citric acid does not affect salivary cortisol levels.

    Science.gov (United States)

    Brorsson, Camilla; Dahlqvist, Per; Nilsson, Leif; Naredi, Silvana

    2014-08-01

    In critically ill patients with hypotension, who respond poorly to fluids and vasoactive drugs, cortisol insufficiency may be suspected. In serum over 90% of cortisol is protein-bound, thus routine measures of total serum cortisol may yield 'false lows' due to hypoproteinaemia. Thus, the occurrence of cortisol insufficiency could be overestimated in critically ill patients. Salivary cortisol can be used as a surrogate for free serum cortisol, but in critically ill patients saliva production is decreased, and insufficient volume of saliva for analysis is a common problem. The aim of this study was to investigate if a cotton-tipped applicator with glycerine and citric acid could be used for saliva stimulation without affecting salivary cortisol levels. Prospective, observational study. Thirty-six volunteers (six males, 30 females), age 49 ± 9 years, without known oral mucus membrane rupture in the mouth. Forty-two pairs of saliva samples (22 paired morning samples, 20 paired evening samples) were obtained before and after saliva stimulation with glycerine and citric acid. Salivary cortisol was analysed using Spectria Cortisol RIA (Orion Diagnostica, Finland). The paired samples correlated significantly (P citric acid did not significantly influence salivary cortisol levels in healthy volunteers. This indicates that salivary cortisol measurement after saliva stimulation may be a useful complement when evaluating cortisol status in critically ill patients. © 2014 John Wiley & Sons Ltd.

  1. The Y4-RNA fragment, a potential diagnostic marker, exists in saliva

    Directory of Open Access Journals (Sweden)

    Tatsuya Ishikawa

    2017-06-01

    Full Text Available The 94-nt full-length Y4-RNA is thought to have roles in the initiation of DNA replication and RNA quality control. Although its 31/32-nt fragment also exists abundantly in plasma, little is known about its physiological role. Since the 31/32-nt Y4-RNA fragment in sera is reported to be more abundant in patients with coronary artery disease than healthy persons, the fragment may have a potential for a diagnostic and/or prognostic biomarker for some diseases regardless of its functionality. As a step toward further investigation of its potential utility, we examined if the 31/32-nt Y4-RNA fragment also exists in saliva that can be obtained noninvasively, and showed that, in addition to the 31/32-nt fragment, 14- and 11-nt Y4-RNA fragments are present in all saliva RNA samples from four healthy persons. We established a PCR method to accurately quantitate the amount of the 31/32-nt Y4-RNA fragment, and estimated its amount in saliva of healthy persons to be 0.06 ± 0.04 fmol per nanogram of saliva RNA. We also tried to develop an easier quantitation method using a DNA molecular beacon. Keywords: Y4-RNA fragment, Saliva RNA, Diagnostic/prognostic marker, Next-generation sequencing, RT-PCR, Molecular beacon

  2. Compartmentalized Self-Replication under fast PCR cycling conditions yields Taq DNA polymerase mutants with increased DNA-binding affinity and blood resistance

    Directory of Open Access Journals (Sweden)

    Bahram eArezi

    2014-08-01

    Full Text Available Faster-cycling PCR formulations, protocols, and instruments have been developed to address the need for increased throughput and shorter turn-around times for PCR-based assays. Although run times can be cut by up to 50%, shorter cycle times have been correlated with lower detection sensitivity and increased variability. To address these concerns, we applied Compartmentalized Self Replication (CSR to evolve faster-cycling mutants of Taq DNA polymerase. After five rounds of selection using progressively shorter PCR extension times, individual mutations identified in the fastest-cycling clones were randomly combined using ligation-based multi-site mutagenesis. The best-performing combinatorial mutants exhibit 35- to 90-fold higher affinity (lower Kd for primed template and a moderate (2-fold increase in extension rate compared to wild-type Taq. Further characterization revealed that CSR-selected mutations provide increased resistance to inhibitors, and most notably, enable direct amplification from up to 65% whole blood. We discuss the contribution of individual mutations to fast-cycling and blood-resistant phenotypes.

  3. Repeated measures study of weekly and daily cytomegalovirus shedding patterns in saliva and urine of healthy cytomegalovirus-seropositive children.

    Science.gov (United States)

    Cannon, Michael J; Stowell, Jennifer D; Clark, Rebekah; Dollard, Philip R; Johnson, Delaney; Mask, Karen; Stover, Cynthia; Wu, Karen; Amin, Minal; Hendley, Will; Guo, Jing; Schmid, D Scott; Dollard, Sheila C

    2014-11-13

    To better understand potential transmission risks from contact with the body fluids of children, we monitored the presence and amount of CMV shedding over time in healthy CMV-seropositive children. Through screening we identified 36 children from the Atlanta, Georgia area who were CMV-seropositive, including 23 who were shedding CMV at the time of screening. Each child received 12 weekly in-home visits at which field workers collected saliva and urine. During the final two weeks, parents also collected saliva and urine daily. Prevalence of shedding was highly correlated with initial shedding status: children shedding at the screening visit had CMV DNA in 84% of follow-up saliva specimens (455/543) and 28% of follow-up urine specimens (151/539); those not shedding at the screening visit had CMV DNA in 16% of follow-up saliva specimens (47/303) and 5% of follow-up urine specimens (16/305). Among positive specimens we found median viral loads of 82,900 copies/mL in saliva and 34,730 copies/mL in urine (P=0.01), while the viral load for the 75th percentile was nearly 1.5 million copies/mL for saliva compared to 86,800 copies/mL for urine. Younger age was significantly associated with higher viral loads, especially for saliva (Pchildren who were shedding at the screening visit were still shedding at least some days during weeks 11 and 12, and median and mean viral loads did not change substantially over time. Healthy CMV-seropositive children can shed CMV for months at high, relatively stable levels. These data suggest that behavioral prevention messages need to address transmission via both saliva and urine, but also need to be informed by the potentially higher risks posed by saliva and by exposures to younger children.

  4. Human saliva as route of inter-human infection for mouse mammary tumor virus.

    Science.gov (United States)

    Mazzanti, Chiara Maria; Lessi, Francesca; Armogida, Ivana; Zavaglia, Katia; Franceschi, Sara; Al Hamad, Mohammad; Roncella, Manuela; Ghilli, Matteo; Boldrini, Antonio; Aretini, Paolo; Fanelli, Giovanni; Marchetti, Ivo; Scatena, Cristian; Hochman, Jacob; Naccarato, Antonio Giuseppe; Bevilacqua, Generoso

    2015-07-30

    Etiology of human breast cancer is unknown, whereas the Mouse Mammary Tumor Virus (MMTV) is recognized as the etiologic agent of mouse mammary carcinoma. Moreover, this experimental model contributed substantially to our understanding of many biological aspects of the human disease. Several data strongly suggest a causative role of MMTV in humans, such as the presence of viral sequences in a high percentage of infiltrating breast carcinoma and in its preinvasive lesions, the production of viral particles in primary cultures of breast cancer, the ability of the virus to infect cells in culture. This paper demonstrates that MMTV is present in human saliva and salivary glands. MMTV presence was investigated by fluorescent PCR, RT-PCR, FISH, immunohistochemistry, and whole transcriptome analysis. Saliva was obtained from newborns, children, adults, and breast cancer patients. The saliva of newborns is MMTV-free, whereas MMTV is present in saliva of children (26.66%), healthy adults (10.60%), and breast cancer patients (57.14% as DNA and 33.9% as RNA). MMTV is also present in 8.10% of salivary glands. RNA-seq analysis performed on saliva of a breast cancer patient demonstrates a high expression of MMTV RNA in comparison to negative controls. The possibility of a contamination by murine DNA was excluded by murine mtDNA and IAP LTR PCR. These findings confirm the presence of MMTV in humans, strongly suggest saliva as route in inter-human infection, and support the hypothesis of a viral origin for human breast carcinoma.

  5. Saliva: A fluid in search of a diagnostic use

    Directory of Open Access Journals (Sweden)

    Jia Liu

    2015-01-01

    Full Text Available Since saliva has been studied for more than 50 years and is relatively easy to collect, it is reasonable to ask why saliva is not in wider use as a diagnostic fluid. Here we discuss the criteria for diagnostic tests for diseases, barriers to use saliva for diagnostic testing, and the possibility of overcoming barriers to acceptance of saliva for diagnosis.

  6. Saliva: A fluid in search of a diagnostic use

    OpenAIRE

    Jia Liu

    2015-01-01

    Since saliva has been studied for more than 50 years and is relatively easy to collect, it is reasonable to ask why saliva is not in wider use as a diagnostic fluid. Here we discuss the criteria for diagnostic tests for diseases, barriers to use saliva for diagnostic testing, and the possibility of overcoming barriers to acceptance of saliva for diagnosis.

  7. Saliva: a reliable sample matrix in bioanalytics.

    Science.gov (United States)

    Gröschl, Michael

    2017-04-01

    Saliva is gaining increasing attention as a bioanalytical sample matrix. Mostly because of the easy and noninvasive collection, it is not only beneficial in endocrinological and behavioral science, but also in pediatrics. Saliva also has the advantage of being the only body fluid which can be collected even during physical exercise, for example, during sportive activities, and there are physiological characteristics that make it superior to serum/plasma or urine for specific scientific questions. This review provides an insight into the physiology of saliva formation, explaining how certain compounds enter this bodily fluid, and gives advice for collection, storage and analytical methods. Finally, it presents a number of reliable and proven applications for saliva analysis from scientific fields including endocrinology, sports medicine, forensics and immunology.

  8. Zika Probably Not Spread Through Saliva: Study

    Science.gov (United States)

    ... page: https://medlineplus.gov/news/fullstory_167531.html Zika Probably Not Spread Through Saliva: Study Research with ... HealthDay News) -- Scientists have some interesting news about Zika: You're unlikely to get the virus from ...

  9. An inhibitor of phospholipase D in saliva

    Science.gov (United States)

    Dawson, Rex M. C.; Hemington, Norma

    1974-01-01

    1. Bovine, dog and human saliva contain substances which inhibit the soluble phospholipase D present in grass leaf or celery stalk. 2. The inhibitor in bovine saliva is of high molecular weight and exhibits considerable stability to heat, acids and alkalis. 3. The inhibitor has been purified free from salivary mucoprotein. 4. It is suggested that the inhibitor could protect the upper alimentary tract of a herbage-eating animal from the necrotic action of phospholipase D. PMID:4376946

  10. Deteksi HBsAg dan HBeAg dalam Saliva Pengidap Virus Hepatitis B

    Directory of Open Access Journals (Sweden)

    Riemawati A. Lesmana

    2015-10-01

    Full Text Available Transmission of hepatitis B virus (HBV via blood or its product has been well established. However, body fluids like urine and saliva may also contain HBV. A complete HBV consists of HBsAg, HBeAg, HBcAg dan partikel DNA. Hepatitis B carrier is detected by the presence of serologic marker HBsAg while the ongoing viral replication or infectivity is diagnosed by the presence of HBeAg or DNA particle. Meanwhile dentists will often contact with the saliva of their patients in daily practice. This cross-sectional study was carried out to assess the infectivity of the saliva of HBV carriers. During a 10 month period (August 1994 - May 1995 detection of HBsAg and HBeAg in blood and saliva were done in 97 HBV carriers using the ELISA method (Enzyme Linked Immunosorbent Assay. Of 97 HBV carriers both positive gor HBsAG in blood were found 56 (Group I and positive HBsAg and negativa HBeAg in the other 41 (Group II. Examination of the saliva of HBV carriers in Group I showed positive HBsAg as well as HBeAg in 48 (85,7%, only positive for HBsAg in 5 (10,7% and both negative for HBsAg and HBeAg in the other 2 (3,6% where as in Group II positive for both HBsAg and HBeAG in the remaining 10 (24,4%. In conclusion, the majority of highly infectious hepatitis B carriers do also have infectious saliva which could be an important source of infection and transmission of the virus in the field of dentistry.

  11. Proteomic profiling of cereal aphid saliva reveals both ubiquitous and adaptive secreted proteins.

    Directory of Open Access Journals (Sweden)

    Sohail A K Rao

    Full Text Available The secreted salivary proteins from two cereal aphid species, Sitobion avenae and Metopolophium dirhodum, were collected from artificial diets and analysed by tandem mass spectrometry. Protein identification was performed by searching MS data against the official protein set from the current pea aphid (Acyrthosiphon pisum genome assembly and revealed 12 and 7 proteins in the saliva of S. avenae and M. dirhodum, respectively. When combined with a comparable dataset from A. pisum, only three individual proteins were common to all the aphid species; two paralogues of the GMC oxidoreductase family (glucose dehydrogenase; GLD and ACYPI009881, an aphid specific protein previously identified as a putative component of the salivary sheath. Antibodies were designed from translated protein sequences obtained from partial cDNA sequences for ACYPI009881 and both saliva associated GLDs. The antibodies detected all parent proteins in secreted saliva from the three aphid species, but could only detect ACYPI009881, and not saliva associated GLDs, in protein extractions from the salivary glands. This result was confirmed by immunohistochemistry using whole and sectioned salivary glands, and in addition, localised ACYPI009881 to specific cell types within the principal salivary gland. The implications of these findings for the origin of salivary components and the putative role of the proteins identified are discussed in the context of our limited understanding of the functional relationship between aphid saliva and the plants they feed on. The mass spectrometry data have been deposited to the ProteomeXchange and can be accessed under the identifier PXD000113.

  12. Proteomic profiling of cereal aphid saliva reveals both ubiquitous and adaptive secreted proteins.

    Science.gov (United States)

    Rao, Sohail A K; Carolan, James C; Wilkinson, Tom L

    2013-01-01

    The secreted salivary proteins from two cereal aphid species, Sitobion avenae and Metopolophium dirhodum, were collected from artificial diets and analysed by tandem mass spectrometry. Protein identification was performed by searching MS data against the official protein set from the current pea aphid (Acyrthosiphon pisum) genome assembly and revealed 12 and 7 proteins in the saliva of S. avenae and M. dirhodum, respectively. When combined with a comparable dataset from A. pisum, only three individual proteins were common to all the aphid species; two paralogues of the GMC oxidoreductase family (glucose dehydrogenase; GLD) and ACYPI009881, an aphid specific protein previously identified as a putative component of the salivary sheath. Antibodies were designed from translated protein sequences obtained from partial cDNA sequences for ACYPI009881 and both saliva associated GLDs. The antibodies detected all parent proteins in secreted saliva from the three aphid species, but could only detect ACYPI009881, and not saliva associated GLDs, in protein extractions from the salivary glands. This result was confirmed by immunohistochemistry using whole and sectioned salivary glands, and in addition, localised ACYPI009881 to specific cell types within the principal salivary gland. The implications of these findings for the origin of salivary components and the putative role of the proteins identified are discussed in the context of our limited understanding of the functional relationship between aphid saliva and the plants they feed on. The mass spectrometry data have been deposited to the ProteomeXchange and can be accessed under the identifier PXD000113.

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

    OpenAIRE

    Elham Farokh-Gisour,; Hadis Fatholah Nejhad; Hamid Reza Poureslami

    2016-01-01

    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...

  14. Detection of hepatitis C virus RNA in saliva samples from patients with seric anti-HCV antibodies

    Directory of Open Access Journals (Sweden)

    Patrícia L. Gonçalves

    Full Text Available We examined the frequency of HCV-RNA in saliva samples from anti-HCV positive patients. Both plasma and saliva samples from 39 HCV patients (13 with normal liver enzymes, 19 with abnormal liver enzymes and 13 with cirrhosis were investigated. Stimulated saliva and fresh plasma were centrifuged (900 x g,10 min and stored at -70ºC, after the addition of guanidine isothiocyanate RNA extraction buffer. HCV-RNA was detected by RT- nested-PCR (amplification of HCV-cDNA for two rounds, using HCV primers 939/209 and 940/211. HCV genotyping was carried out by RFLP (using Mva I and Hinf 1 or Hae III and Rsa I restriction enzymes. Thirty-two out of 39 (82%; 95% CI=70-94% anti-HCV-positive patients had HCV-RNA in plasma samples. Eight out of 39 (20.5%; 95% CI=6.6-34.4% had HCV-RNA in the saliva. The HCV genotype in saliva samples from these patients matched the genotype found for plasma HCV-RNA. No significant correlation between the presence of HCV and either age, gender, HCV genotype or any risk factor for HCV infection was found. The observed prevalence (20.5% of anti HCV positive patients or 25% of the patients with HCV-RNA in plasma was lower than that previously reported from other countries. The low frequency of HCV-RNA in saliva samples observed in our study may be due to the use of cell-free saliva. Other authors reporting higher frequencies of HCV-RNA in saliva used whole saliva, without centrifugation.

  15. HCV clearance patterns in saliva and serum of patients with chronic HCV infection under interferon plus ribavirin therapy.

    Science.gov (United States)

    Diz Dios, P; Castro, A; Rodríguez, I; Reforma, N G; Castro, M; Eirea, M; Hermida, M

    2005-05-01

    Hepatitis C virus (HCV)-RNA is often present in saliva of HCV-infected patients, with plasma viral load being the only known predictable factor. Interferon plus ribavirin therapy yields a sustained reduction in HCV viremia. This study aimed to assess the presence of HCV in saliva and serum specimens from patients undergoing this combination therapy (CT). Paired serum and saliva specimens were collected from 44 chronic HCV-infected patients at basal time, 4 and 12 weeks after CT onset, at the end of treatment and 6 months latter. Serum HCV-RNA levels were determined by the polymerase chain reaction (PCR) Amplicor system. Presence of HCV-RNA in saliva was tested by a highly sensitive non-commercialized nested-PCR. The HCV-RNA was detected in 26 saliva specimens at basal time (59.1%). In 34.1% of cases, a concordance viral clearance pattern in serum and saliva was observed in both responders (pattern 1a) and non-responders (pattern 1b). In pattern 2 (13.6% of cases), HCV was detected longer during CT in serum than in saliva (pattern 2a) or in saliva than in serum (pattern 2b). In 11.3% of patients, viral clearance was corroborated either in their serum (pattern 3a) or in their saliva (pattern 3b), but not in both fluids. Of the eight primary responders with 1a clearance pattern, seven were sustained responders. None of the patients with 2a clearance pattern was a sustained responder. Of the two primary responders showing the 3b salivary pattern, one had already relapsed in the first 6 months of follow up. The present results suggest that the monitoring of salivary levels of HCV would be a helpful means of determining sustained antiviral effects of interferon and ribavirin in the treatment of HCV disease.

  16. 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...... mellem fluoridkoncentrationerne i biofilmsediment og fordeling af caries mellem regioner. Forskere anbefales at være opmærksomme på de intra-orale forskelle i fluoridkoncentrationer mellem regioner samt sikre balancering af effekten af region, når der indsamles dental biofilm til fluoridanalyse. I det...... 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...

  17. The use of saliva as a practical and feasible alternative to urine in large-scale screening for congenital cytomegalovirus infection increasesinclusion and detection rates

    Directory of Open Access Journals (Sweden)

    Emanuelle Santos de Carvalho Cardoso

    2015-04-01

    Full Text Available INTRODUCTION: Although urine is considered the gold-standard material for the detection of congenital cytomegalovirus (CMV infection, it can be difficult to obtain in newborns. The aim of this study was to compare the efficiency of detection of congenital CMV infection in saliva and urine samples. METHODS: One thousand newborns were included in the study. Congenital cytomegalovirus deoxyribonucleic acid (DNA was detected by polymerase chain reaction (PCR. RESULTS: Saliva samples were obtained from all the newborns, whereas urine collection was successful in only 333 cases. There was no statistically significant difference between the use of saliva alone or saliva and urine collected simultaneously for the detection of CMV infection. CONCLUSIONS: Saliva samples can be used in large-scale neonatal screening for CMV infection.

  18. 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.

  19. Development of a Rapid Point-of-Use DNA Test for the Screening of Genuity® Roundup Ready 2 Yield® Soybean in Seed Samples

    Directory of Open Access Journals (Sweden)

    Dilip Chandu

    2016-01-01

    Full Text Available Testing for the presence of genetically modified material in seed samples is of critical importance for all stakeholders in the agricultural industry, including growers, seed manufacturers, and regulatory bodies. While rapid antibody-based testing for the transgenic protein has fulfilled this need in the past, the introduction of new variants of a given transgene demands new diagnostic regimen that allows distinguishing different traits at the nucleic acid level. Although such molecular tests can be performed by PCR in the laboratory, their requirement for expensive equipment and sophisticated operation have prevented its uptake in point-of-use applications. A recently developed isothermal DNA amplification technique, recombinase polymerase amplification (RPA, combines simple sample preparation and amplification work-flow procedures with the use of minimal detection equipment in real time. Here, we report the development of a highly sensitive and specific RPA-based detection system for Genuity Roundup Ready 2 Yield (RR2Y material in soybean (Glycine max seed samples and present the results of studies applying the method in both laboratory and field-type settings.

  20. Saliva: A tool in assessing glucose levels in Diabetes Mellitus

    National Research Council Canada - National Science Library

    Satish, B N V S; Srikala, P; Maharudrappa, B; Awanti, Sharanabasappa M; Kumar, Prashant; Hugar, Deepa

    2014-01-01

    .... The aim of the present study was undertaken to correlate the glucose levels in saliva and blood of diabetic and healthy non diabetic individuals and to determine the efficacy of saliva as a diagnostic...

  1. Streptococcus pneumoniae in saliva of Dutch primary school children

    NARCIS (Netherlands)

    Wyllie, Anne L.; Chu, Mei Ling J. N.; Schellens, Mariëlle H. B.; van Engelsdorp Gastelaars, Jody; Jansen, Marc D.; van der Ende, Arie; Bogaert, Debby; Sanders, Elisabeth A. M.; Trzciński, Krzysztof

    2014-01-01

    While nasopharyngeal sampling is the gold standard for the detection of Streptococcus pneumoniae carriage, historically seen, saliva sampling also seems highly sensitive for pneumococcal detection. We investigated S. pneumoniae carriage in saliva from fifty schoolchildren by conventional and

  2. DNA research in forensic dentistry.

    Science.gov (United States)

    Lijnen, I; Willems, G

    2001-11-01

    DNA analysis has recently been introduced to forensic dentistry and is now frequently used in identifying individuals or determining the origin of certain tissues. This review reports on teeth and saliva as a source of DNA. Not only the quantity of DNA available for the laboratory is important, but also the quality and purity. Teeth are resistant against extreme circumstances such as temperature, humidity and acidity, which is an important advantage in DNA analysis. Furthermore, an abundance of DNA can be extracted from teeth. Saliva can be obtained in a simple, painless and non-radical way. The double swab method is very effective; DNA recovery is significantly higher with the double swab method compared to the single swab or filter paper method. This review reports on the different techniques used to extract DNA from teeth and saliva, as well as DNA analysis of these samples. The usefulness and advantages of the double swab method for saliva, cryogenic grinding for teeth and the chelex extraction and polymerase chain reaction for both types of samples is also described. DNA analysis has proven its value in forensic dentistry, but ethical and juridical considerations are still a matter of debate and criticism.

  3. [Study on mobile phone enabled wireless detection of saliva glucose].

    Science.gov (United States)

    Li, Jingjing; Yu, Yang; Lu, Yongqiang; Liu, Jing

    2011-09-01

    In this study, based on the correlation between the blood and saliva glucose, we proposed and developed a new conceptual method of using mobile phone to measure wirelessly the glucose concentration in saliva. According to the experiments on simulated saliva, the new system could draw, display, store and carry out calculation on the correlation curves between saliva glucose and electrical parameters. This demonstrates the feasibility and bright future of the new technique.

  4. Re-evaluation of saliva for monitoring theophylline concentrations.

    OpenAIRE

    Rylance, G W; Beswick, D T; Cullen, R. E.; Roberts, D G

    1985-01-01

    Variability of the mixed saliva/plasma theophylline relation was examined in seven children aged 2 to 13 years. Good correlation between plasma and saliva concentrations was found, but on the three occasions there was considerable inter- and intrapatient variability. There was no significant or consistent relation between unstimulated and stimulated saliva concentrations or between saliva concentrations and sample volumes. Plasma theophylline concentrations cannot be predicted accurately from...

  5. Influence of Individual Saliva Secretion on Fluoride Bioavailability

    OpenAIRE

    E. A. Naumova; Gaengler, P.; Zimmer, S.; Arnold, W. H.

    2010-01-01

    The aim of this preliminary investigation was to compare the individual saliva secretion rate with the fluoride bioavailability in saliva after using sodium fluoride and amine fluoride. Methods: To assess oral fluoride kinetics 10 highly trained volunteers brushed their teeth with one of the formulations and saliva was collected. The amount of saliva was measured, and the fluoride content was determined. Data underwent statistical analysis using the Mann-Whitney-U test and Pearson correlation...

  6. Rapid extraction and preservation of genomic DNA from human samples.

    Science.gov (United States)

    Kalyanasundaram, D; Kim, J-H; Yeo, W-H; Oh, K; Lee, K-H; Kim, M-H; Ryew, S-M; Ahn, S-G; Gao, D; Cangelosi, G A; Chung, J-H

    2013-02-01

    Simple and rapid extraction of human genomic DNA remains a bottleneck for genome analysis and disease diagnosis. Current methods using microfilters require cumbersome, multiple handling steps in part because salt conditions must be controlled for attraction and elution of DNA in porous silica. We report a novel extraction method of human genomic DNA from buccal swab and saliva samples. DNA is attracted onto a gold-coated microchip by an electric field and capillary action while the captured DNA is eluted by thermal heating at 70 °C. A prototype device was designed to handle four microchips, and a compatible protocol was developed. The extracted DNA using microchips was characterized by qPCR for different sample volumes, using different lengths of PCR amplicon, and nuclear and mitochondrial genes. In comparison with a commercial kit, an equivalent yield of DNA extraction was achieved with fewer steps. Room-temperature preservation for 1 month was demonstrated for captured DNA, facilitating straightforward collection, delivery, and handling of genomic DNA in an environment-friendly protocol.

  7. A proteomic approach to porcine saliva.

    Science.gov (United States)

    Gutiérrez, Ana M; Cerón, José J; Fuentes-Rubio, María; Tecles, Fernando; Beeley, Josie A

    2014-02-01

    This paper reviews recent progress in salivary animal proteomics, with special reference to the porcine proteome. Until fairly recently, most studies on saliva as a diagnostic fluid have focused on humans, primates and rodents, and the development of salivary analysis in monitoring health in farm animals including pigs has received only limited consideration. The porcine salivary proteome has been characterised by 2D-electrophoresis followed by mass spectrometry. Major and minor proteins have been identified. The use of saliva as a non-invasive biological fluid in monitoring health and disease in pigs will be reviewed, together with the potential use of proteomics for the development of biomarkers. In this review, methods of collection and the composition of porcine saliva will be considered, together with saliva handling and analysis. The overall findings indicate that there is considerable potential for the development of salivary analysis as a non-invasive diagnostic fluid in the pig, and that it offers advantages over other body fluids in this animal.

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

    Directory of Open Access Journals (Sweden)

    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

  9. Saliva as research material: Biochemical, physicochemical and practical aspects

    NARCIS (Netherlands)

    Schipper, R.G.; Silletti, E.; Vingerhoeds, M.H.

    2007-01-01

    Whole saliva is a complex mixture of proteins and other molecules which originate from several sources. The biochemical and physicochemical properties of saliva contribute to the numerous functions of saliva in, e.g., speech, maintaining oral and general health, and food processing. Interest in

  10. RHEOLOGICAL ASPECTS OF MUCIN-CONTAINING SOLUTIONS AND SALIVA SUBSTITUTES

    NARCIS (Netherlands)

    HOLTERMAN, HJ; WATERMAN, HA; BLOM, C; SGRAVENMADE, FJ; Mellema, J.

    1992-01-01

    In this study rheological properties of aqueous solutions of mucin, albumin and mucin-albumin have been investigated in search for saliva substitutes. They were compared with commercially available saliva substitutes on the one hand and natural human saliva on the other hand. For the latter a few

  11. Predictors of mother and child DNA yields in buccal cell samples collected in pediatric cancer epidemiologic studies: a report from the Children's Oncology group

    National Research Council Canada - National Science Library

    Poynter, Jenny N; Ross, Julie A; Hooten, Anthony J; Langer, Erica; Blommer, Crystal; Spector, Logan G

    2013-01-01

    ...; however, DNA collection in young children poses additional challenges. Here, we have evaluated predictors of DNA quantity in buccal cells collected for population-based studies of infant leukemia (N...

  12. Saliva as a diagnostic fluid. Literature review.

    Science.gov (United States)

    Martí-Álamo, Silvia; Mancheño-Franch, Aisha; Marzal-Gamarra, Cristina; Carlos-Fabuel, Laura

    2012-10-01

    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 amount of desquamated epithelial cells, microorganisms and remnants of food and drink; it is essential to cleanse and refine the saliva samples to remove any external elements. Immediate processing of the sample is recommended in order to avoid decomposition, where this is not possible, the sample may be stored at -80ºC. Salivary analysis - much the same as blood analysis - aims to identify diverse medication or indications of certain diseases while providing a relatively simple tool for both early diagnosis and monitoring various irregularities. The practicalities of salivary analysis have been studied in fields such as: viral and bacterial infections, autoimmune diseases (like Sjögren's syndrome and cɶliac disease), endocrinopathies (such as Cushing's syndrome), oncology (early diagnosis of breast, lung and stomach carcinoma and oral squamous cell carcinoma), stress assessment, medication detection and forensic science among others. It is hoped that salivary analysis, with the help of current technological advances, will be valued much more highly in the near future. There still remain contradictory results with respect to analytic markers, which is why further studies into wider-ranging samples are fundamental to prove its viability. Key words:Saliva, biomarkers, early diagnosis.

  13. 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 (diagnostic accuracy was poor (AUC = 0.77-0.78) for all body fluids evaluated. Strong agreement was observed between Sosm methods (Expectoration: 61 ± 10 mmol/kg, Salivette: 61 ± 8 mmol/kg, p > 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.

  14. Saliva as a diagnostic fluid. Literature review

    Science.gov (United States)

    Mancheño-Franch, Aisha; Marzal-Gamarra, Cristina; Carlos-Fabuel, Laura

    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 amount of desquamated epithelial cells, microorganisms and remnants of food and drink; it is essential to cleanse and refine the saliva samples to remove any external elements. Immediate processing of the sample is recommended in order to avoid decomposition, where this is not possible, the sample may be stored at -80ºC. Salivary analysis – much the same as blood analysis – aims to identify diverse medication or indications of certain diseases while providing a relatively simple tool for both early diagnosis and monitoring various irregularities. The practicalities of salivary analysis have been studied in fields such as: viral and bacterial infections, autoimmune diseases (like Sjögren’s syndrome and cɶliac disease), endocrinopathies (such as Cushing’s syndrome), oncology (early diagnosis of breast, lung and stomach carcinoma and oral squamous cell carcinoma), stress assessment, medication detection and forensic science among others. It is hoped that salivary analysis, with the help of current technological advances, will be valued much more highly in the near future. There still remain contradictory results with respect to analytic markers, which is why further studies into wider-ranging samples are fundamental to prove its viability. Key words:Saliva, biomarkers, early diagnosis. PMID:24558562

  15. Biomonitorization of cadmium, chromium, manganese, nickel and lead in whole blood, urine, axillary hair and saliva in an occupationally exposed population

    Energy Technology Data Exchange (ETDEWEB)

    Gil Fernando, E-mail: fgil@ugr.es [Department of Legal Medicine and Toxicology, University of Granada, School of Medicine (Spain); Hernandez, Antonio F. [Department of Legal Medicine and Toxicology, University of Granada, School of Medicine (Spain); Marquez, Claudia [Internal Resident in Occupational Medicine, School of Occupational Medicine of University of Granada (Spain); Femia, Pedro [Department of Statistics, University of Granada, School of Medicine (Spain); Olmedo, Pablo; Lopez-Guarnido, Olga; Pla, Antonio [Department of Legal Medicine and Toxicology, University of Granada, School of Medicine (Spain)

    2011-02-15

    Heavy metal contamination from occupational origin is a cause for concern because of its potential accumulation in the environment and in living organisms leading to long term toxic effects. This study was aimed to assess Cd, Cr, Mn, Ni and Pb levels in whole blood, urine, axillary hair and saliva from 178 individuals with occupational exposure to heavy metals. Levels of metal compounds were determined by atomic absorption spectrometry. We collected information on occupation, lifestyle habits and food intake by questionnaire. Multiple linear regression analyses for metal ion concentration in whole blood, urine, axillary hair and saliva were adjusted for age, gender, smoking and alcohol consumption, lifetime workplace exposure, residence area and food habits. Overall, blood and urine median concentrations found for the five metals analyzed do not exceed biological exposure indexes, so that they are very similar to a non-occupationally exposed population. Toxicokinetic differences may account for the lack of correlations found for metal levels in hair and saliva with those in blood or urine. For those heavy metals showing higher median levels in blood with respect to hair (Cd, Mn and Pb) indicating lesser hair incorporation from blood, the lifetime working experience was inversely correlated with their hair levels. The longer the lifetime working experience in industrial environments, the higher the Mn and Ni concentration in saliva. Axillary hair and saliva may be used as additional and/or alternative samples to blood or urine for biomonitoring hair Mn, and saliva Ni in subjects with occupational exposure. - Research Highlights: {yields} Metal levels in workers were similar to an occupationally non-exposed population. {yields} Metal levels in blood and urine were below recommended reference values. {yields} A lack of correlation was observed between metal levels in blood and saliva. {yields} Toxicokinetic differences may account for the lack of correlations observed

  16. Surface-enhanced Raman spectroscopy of saliva proteins for the noninvasive differentiation of benign and malignant breast tumors

    Science.gov (United States)

    Feng, Shangyuan; Huang, Shaohua; Lin, Duo; Chen, Guannan; Xu, Yuanji; Li, Yongzeng; Huang, Zufang; Pan, Jianji; Chen, Rong; Zeng, Haishan

    2015-01-01

    The capability of saliva protein analysis, based on membrane protein purification and surface-enhanced Raman spectroscopy (SERS), for detecting benign and malignant breast tumors is presented in this paper. A total of 97 SERS spectra from purified saliva proteins were acquired from samples obtained from three groups: 33 healthy subjects; 33 patients with benign breast tumors; and 31 patients with malignant breast tumors. Subtle but discernible changes in the mean SERS spectra of the three groups were observed. Tentative assignments of the saliva protein SERS spectra demonstrated that benign and malignant breast tumors led to several specific biomolecular changes of the saliva proteins. Multiclass partial least squares–discriminant analysis was utilized to analyze and classify the saliva protein SERS spectra from healthy subjects, benign breast tumor patients, and malignant breast tumor patients, yielding diagnostic sensitivities of 75.75%, 72.73%, and 74.19%, as well as specificities of 93.75%, 81.25%, and 86.36%, respectively. The results from this exploratory work demonstrate that saliva protein SERS analysis combined with partial least squares–discriminant analysis diagnostic algorithms has great potential for the noninvasive and label-free detection of breast cancer. PMID:25609959

  17. Canine-specific STR typing of saliva traces on dog bite wounds.

    Science.gov (United States)

    Eichmann, Cordula; Berger, Burkhard; Reinhold, Maximilian; Lutz, Martin; Parson, Walther

    2004-12-01

    Forensic investigations in dog attacks usually involve the examination of bite marks and toothprints, the dog's stomach and pathological methods. For identification of the offending dog we evaluated canine STR typing of saliva traces on dog bite marks. The specificity of 15 canine-specific STRs was tested on human-canine DNA mixtures prior to an applied study in which 52 cases of dog bites were investigated. The first-aid wound bandages as well as swab samples from the surrounding area of the wound were used for DNA analyses. Generally, it was possible to obtain a canine-specific STR profile from the dog's saliva left on the wound area, even when high background of human DNA was present (blood). Interestingly, we found canine STR typing to be more successful when the bandages and swabs showed high amounts of human blood, i.e. when the dog bite was severe. Canine saliva was then sometimes visible as white-coloured secretion on the human blood surface. Less severe bite cases, which did not result in bleeding wounds, showed less success in obtaining useful STR results, probably due to the fact that the surface of the wounds may have been treated before the victims consulted medical aid which therefore removed the canine cells.

  18. Viral latency in blood and saliva of simian foamy virus-infected humans.

    Directory of Open Access Journals (Sweden)

    Rejane Rua

    Full Text Available Simian foamy viruses (SFV are widespread retroviruses among non-human primates (NHP. SFV actively replicate in the oral cavity and can be transmitted to humans through NHP bites, giving rise to a persistent infection. We aimed at studying the natural history of SFV infection in human. We have analyzed viral load and gene expression in 14 hunters from Cameroon previously shown to be infected with a gorilla SFV strain. Viral DNA could be detected by quantitative polymerase chain reaction (q-PCR targeting the pol-in region, in most samples of peripheral blood mononuclear cells (PBMCs (7.1 ± 6.0 SFV DNA copies/105 PBMCs and saliva (2.4 ± 4.3 SFV DNA copies/105 cells derived from the hunters. However, quantitative real-time reverse-transcription polymerase chain reaction (RT-qPCR revealed the absence of SFV viral gene expression in both PBMCs and saliva, suggesting that SFV was latent in the human samples. Our study demonstrates that a latent infection can occur in humans and persist for years, both in PBMCs and saliva. Such a scenario may contribute to the putative lack of secondary human-to-human transmissions of SFV.

  19. Non-coding RNAs in saliva: emerging biomarkers for molecular diagnostics.

    Science.gov (United States)

    Majem, Blanca; Rigau, Marina; Reventós, Jaume; Wong, David T

    2015-04-17

    Saliva is a complex body fluid that comprises secretions from the major and minor salivary glands, which are extensively supplied by blood. Therefore, molecules such as proteins, DNA, RNA, etc., present in plasma could be also present in saliva. Many studies have reported that saliva body fluid can be useful for discriminating several oral diseases, but also systemic diseases including cancer. Most of these studies revealed messenger RNA (mRNA) and proteomic biomarker signatures rather than specific non-coding RNA (ncRNA) profiles. NcRNAs are emerging as new regulators of diverse biological functions, playing an important role in oncogenesis and tumor progression. Indeed, the small size of these molecules makes them very stable in different body fluids and not as susceptible as mRNAs to degradation by ribonucleases (RNases). Therefore, the development of a non-invasive salivary test, based on ncRNAs profiles, could have a significant applicability to clinical practice, not only by reducing the cost of the health system, but also by benefitting the patient. Here, we summarize the current status and clinical implications of the ncRNAs present in human saliva as a source of biological information.

  20. Non-Coding RNAs in Saliva: Emerging Biomarkers for Molecular Diagnostics

    Science.gov (United States)

    Majem, Blanca; Rigau, Marina; Reventós, Jaume; Wong, David T.

    2015-01-01

    Saliva is a complex body fluid that comprises secretions from the major and minor salivary glands, which are extensively supplied by blood. Therefore, molecules such as proteins, DNA, RNA, etc., present in plasma could be also present in saliva. Many studies have reported that saliva body fluid can be useful for discriminating several oral diseases, but also systemic diseases including cancer. Most of these studies revealed messenger RNA (mRNA) and proteomic biomarker signatures rather than specific non-coding RNA (ncRNA) profiles. NcRNAs are emerging as new regulators of diverse biological functions, playing an important role in oncogenesis and tumor progression. Indeed, the small size of these molecules makes them very stable in different body fluids and not as susceptible as mRNAs to degradation by ribonucleases (RNases). Therefore, the development of a non-invasive salivary test, based on ncRNAs profiles, could have a significant applicability to clinical practice, not only by reducing the cost of the health system, but also by benefitting the patient. Here, we summarize the current status and clinical implications of the ncRNAs present in human saliva as a source of biological information. PMID:25898412

  1. [THE MOLECULAR GENETIC CHARACTERISTIC OF SPECIES CONTENT OF SALIVA AND GINGIVAL RECESS UNDER PERIODONTITIS].

    Science.gov (United States)

    Tamarova, E R; Baimiev, A Kh; Shvetz, K Yu; Mavzyutov, A R

    2015-12-01

    The examination was carried out of samplings of 110 patients with periodontitis (observation group) and 60 patients without pathology of periodont (comparison group). The polymerase chain reaction was used to analyze samples of saliva and contents of periodontal recesses for detecting species-specific DNA fragments of Porphymmonas gigngivalis, Streptococcus macacae, S. mutans, S. oralis, S. salivarius, S. sangis, S. sobrinus, Treponema denticola. In patients with periodontitis S. mutans, S. oralis S. sobrinus were reliably more often detected in the content of periodontal recesses and S. mutans, S. sobrinus i in saliva. In the observation group the rate of detection of association S. mutans--S. oralis--S. sangis--S. sobrinus was significantly exceeded (up to 15.6%, X2 = 9.1, p = 0.004). In ten days of effective treatment of periodontitis reliable decreastng of rate of detection of S. wasoralis, S. sobrinus was observed in contents of periodontal recesses but not in of saliva. The detection of S.sobrinus using technique of polymerase chain reaction in contents of periodontal recesses and/or saliva of patients with periodontitis has a diagnostic value. The detection of S.sobrinus in contents of periodontal recesses is significant both in monoculture and in association S. mutans--S. oralis--S. sangis--S.sobrinus. The absence of S. sobrinus in contents of periodontal recesses testifies effectiveness of treatment of main disease (periodontitis).

  2. Protein Quality Assessment on Saliva Samples for Biobanking Purposes.

    Science.gov (United States)

    Rosa, Nuno; Marques, Jéssica; Esteves, Eduardo; Fernandes, Mónica; Mendes, Vera M; Afonso, Ângela; Dias, Sérgio; Pereira, Joaquim Polido; Manadas, Bruno; Correia, Maria José; Barros, Marlene

    2016-08-01

    Biobank saliva sample quality depends on specific criteria applied to collection, processing, and storage. In spite of the growing interest in saliva as a diagnostic fluid, few biobanks currently store large collections of such samples. The development of a standard operating procedure (SOP) for saliva collection and quality control is fundamental for the establishment of a new saliva biobank, which stores samples to be made available to the saliva research community. Different collection methods were tested regarding total volume of protein obtained, protein content, and protein profiles, and the results were used to choose the best method for protein studies. Furthermore, the impact of the circadian variability and inter- and intraindividual differences, as well as the saliva sample stability at room temperature, were also evaluated. Considering our results, a sublingual cotton roll method for saliva collection proved to produce saliva with the best characteristics and should be applied in the morning, whenever possible. In addition, there is more variability in salivary proteins between individuals than in the same individual for a 5-month period. According to the electrophoretic protein profile, protein stability is guaranteed for 24 hours at room temperature and the protein degradation profile and protein identification were characterized. All this information was used to establish an SOP for saliva collection, processing, and storage in a biobank. We conclude that it is possible to collect saliva using an easy and inexpensive protocol, resulting in saliva samples for protein analysis with sufficient quality for biobanking purposes.

  3. 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.

  4. Factors That Influence the Extensional Rheological Property of Saliva

    Science.gov (United States)

    Vijay, Amrita; Inui, Taichi; Dodds, Michael; Proctor, Gordon; Carpenter, Guy

    2015-01-01

    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. PMID:26305698

  5. Comparison of Saliva Collection Methods in Children with High-Functioning Autism Spectrum Disorders: Acceptability and Recovery of Cortisol

    Science.gov (United States)

    Putnam, Susan K.; Lopata, Christopher; Fox, Jeffery D.; Thomeer, Marcus L.; Rodgers, Jonathan D.; Volker, Martin A.; Lee, Gloria K.; Neilans, Erik G.; Werth, Jilynn

    2012-01-01

    This study compared cortisol concentrations yielded using three saliva collection methods (passive drool, salivette, and sorbette) in both in vitro and in vivo conditions, as well as method acceptability for a sample of children (n = 39) with High Functioning Autism Spectrum Disorders. No cortisol concentration differences were observed between…

  6. Construction and characterization of the Korean whole saliva proteome to determine ethnic differences in human saliva proteome.

    Science.gov (United States)

    Cho, Ha Ra; Kim, Han Sol; Park, Jun Seo; Park, Seung Cheol; Kim, Kwang Pyo; Wood, Troy D; Choi, Yong Seok

    2017-01-01

    As the first step to discover protein disease biomarkers from saliva, global analyses of the saliva proteome have been carried out since the early 2000s, and more than 3,000 proteins have been identified in human saliva. Recently, ethnic differences in the human plasma proteome have been reported, but such corresponding studies on human saliva in this aspect have not been previously reported. Thus, here, in order to determine ethnic differences in the human saliva proteome, a Korean whole saliva (WS) proteome catalogue indexing 480 proteins was built and characterized through nLC-Q-IMS-TOF analyses of WS samples collected from eleven healthy South Korean male adult volunteers for the first time. Identification of 226 distinct Korean WS proteins, not observed in the integrated human saliva protein dataset, and significant gene ontology distribution differences in the Korean WS proteome compared to the integrated human saliva proteome strongly support ethnic differences in the human saliva proteome. Additionally, the potential value of ethnicity-specific human saliva proteins as biomarkers for diseases highly prevalent in that ethnic group was confirmed by finding 35 distinct Korean WS proteins likely to be associated with the top 10 deadliest diseases in South Korea. Finally, the present Korean WS protein list can serve as the first level reference for future proteomic studies including disease biomarker studies on Korean saliva.

  7. Saliva: A diagnostic biomarker of periodontal diseases

    Science.gov (United States)

    Patil, Priti Basgauda; Patil, Basgauda Ramesh

    2011-01-01

    Early detection of disease plays a crucial role in successful therapy. Early diagnosis and management reduces the severity and possible complications of the disease process. To overcome this challenge, medical researchers are devoted to finding molecular disease biomarkers that reveal a hidden lethal threat before the disease becomes complicated. Saliva, an important physiologic fluid, containing a highly complex mixture of substances, is rapidly gaining popularity as a diagnostic tool. Periodontal disease is a chronic disease of the oral cavity comprising a group of inflammatory conditions affecting the supporting structures of the dentition. In the field of periodontology, traditional clinical criteria are often insufficient for determining sites of active disease, for monitoring the response to therapy, or for measuring the degree of susceptibility to future disease progression. Saliva, as a mirror of oral and systemic health, is a valuable source for clinically relevant information because it contains biomarkers specific for the unique physiologic aspects of periodontal diseases. This review highlights the various potentials of saliva as a diagnostic biomarker for periodontal diseases. PMID:22368352

  8. Can saliva replace plasma for the monitoring of methadone?

    Science.gov (United States)

    Shiran, Mohammad Reza; Hassanzadeh-Khayyat, Mohammad; Iqbal, Mohammad Zafar; Lagundoye, Olawale; Seivewright, Nicholas; Lennard, Martin S; Tucker, Geoffrey T; Rostami-Hodjegan, Amin

    2005-10-01

    The aims of this study were to determine the relationship between saliva and plasma methadone concentrations and the influence of variability in saliva pH. Saliva and plasma samples were taken before the daily dose of methadone in 60 patients undergoing methadone maintenance treatment (MMT). Saliva pH was measured immediately after sampling, and concentrations of (RS)-, (R)-, and (S)-methadone in saliva and plasma were assayed by LC/MS. In addition, unbound (R)- and (S)-methadone concentrations were measured in plasma samples by ultrafiltration. Plasma binding and pH differences between plasma and saliva were then used to estimate methadone saliva/plasma ratios and to compare them with observed values. Saliva pH ranged from 5.1 to 7.6 (mean +/- SD, 6.7 +/- 0.5). Plasma and saliva concentrations correlated weakly [(RS)-, r = 0.14, P = 0.007, n = 44; (R)-, r = 0.10, P = 0.04, n = 43; (S)-, r = 0.22, P = 0.002, n = 43], and the mean saliva-to-plasma methadone concentration ratios were 1.1 (+/-1.3 SD), 1.5 (+/-1.5), and 0.8 (+/-0.8), for (RS)-, (R)-, and (S)-methadone, respectively. Corresponding values based on unbound concentrations of methadone in plasma were 21 (+/-20.6, n = 31), 21 (+/-19, n = 34), and 17 (+/-15, n = 36). The salivary concentration-to-dose ratios showed statistically significant but weak inverse correlations with saliva pH [(RS)-, r = 0.27, P < 0.001; (R)-, r = 0.25, P < 0.001; (S)-, r = 0.29, P < 0.001, respectively]. There were significant correlations between predicted and observed saliva/plasma ratios [(RS)-, r = 0.44, P < 0.001, n = 31; (R)-, r = 0.58, P < 0.001, n = 32; (S)-, r = 0.10, P = 0.04, n = 34], but the mean predicted saliva concentrations were about 5 times lower than the mean observed values. The poor correlations between salivary and plasma methadone concentrations observed in this study are partly related to the effect of variable saliva pH. However, saliva pH explained only 10%-36% of the total variation. As a conclusion

  9. Diagnostic potential of saliva: current state and future applications.

    Science.gov (United States)

    Pfaffe, Tina; Cooper-White, Justin; Beyerlein, Peter; Kostner, Karam; Punyadeera, Chamindie

    2011-05-01

    Over the past 10 years, the use of saliva as a diagnostic fluid has gained attention and has become a translational research success story. Some of the current nanotechnologies have been demonstrated to have the analytical sensitivity required for the use of saliva as a diagnostic medium to detect and predict disease progression. However, these technologies have not yet been integrated into current clinical practice and work flow. As a diagnostic fluid, saliva offers advantages over serum because it can be collected noninvasively by individuals with modest training, and it offers a cost-effective approach for the screening of large populations. Gland-specific saliva can also be used for diagnosis of pathology specific to one of the major salivary glands. There is minimal risk of contracting infections during saliva collection, and saliva can be used in clinically challenging situations, such as obtaining samples from children or handicapped or anxious patients, in whom blood sampling could be a difficult act to perform. In this review we highlight the production of and secretion of saliva, the salivary proteome, transportation of biomolecules from blood capillaries to salivary glands, and the diagnostic potential of saliva for use in detection of cardiovascular disease and oral and breast cancers. We also highlight the barriers to application of saliva testing and its advancement in clinical settings. Saliva has the potential to become a first-line diagnostic sample of choice owing to the advancements in detection technologies coupled with combinations of biomolecules with clinical relevance.

  10. Characterization of the activity and stability of amylase from saliva and detergent: Laboratory practicals for studying the activity and stability of amylase from saliva and various commercial detergents

    National Research Council Canada - National Science Library

    Valls, Cristina; Rojas, Cristina; Pujadas, Gerard; Garcia‐Vallve, Santi; Mulero, Miquel

    2012-01-01

    ...) in saliva and detergents. These laboratory practicals are based on the determination of the enzymatic activity of amylase from saliva and different detergents using the Phadebas test (quantitative...

  11. Integrated amplification microarray system in a lateral flow cell for warfarin genotyping from saliva.

    Science.gov (United States)

    Sebastian, Thomas; Cooney, Christopher G; Parker, Jennifer; Qu, Peter; Perov, Alexander; Golova, Julia B; Pozza, Lindsay; Iwasiow, Rafal M; Holmberg, Rebecca

    2014-02-15

    Genetic polymorphisms in the CYP2C9 and VKORC1 genes have been linked to sensitivity of the anticoagulant drug warfarin. The aim of this study is to demonstrate a method for warfarin sensitivity genotyping using gel element microarray technology in a simplified workflow from sample collection to analysis and detection. We developed an integrated amplification microarray system combining PCR amplification, target labeling, and microarray hybridization within a single, closed-amplicon "lateral flow cell" for genotyping three single nucleotide polymorphisms (SNPs) that influence warfarin response. We combined nucleic acid extraction of saliva using the TruTip technology together with the lateral flow cell assay and with fully automated array detection and analysis. The analytical performance of the assay was tested using 20 genotyped human genomic DNA samples and found to be sensitive down to 330 input genomic copies (1 ng). A follow-up pre-clinical evaluation was performed with 65 blinded saliva samples and the genotyping results were in agreement with those determined by bidirectional sequencing. Combined with the use of non-invasive saliva samples, rapid DNA extraction, the lateral flow cell, automatic imaging and data analysis provides a simple and fast sample-to-answer microarray test for warfarin sensitivity genotyping. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. 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

  13. 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

  14. 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.

    Directory of Open Access Journals (Sweden)

    Luciane Almeida Amado Leon

    Full Text Available 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

  15. Analysis of BZLF1 mRNA detection in saliva as a marker for active replication of Epstein-Barr virus.

    Science.gov (United States)

    Fagin, Ursula; Nerbas, Linda; Vogl, Bastian; Jabs, Wolfram J

    2017-06-01

    Monitoring replicative Epstein-Barr virus (EBV) infection still remains a challenge in modern laboratory routine. The immediate-early protein BZLF1 mediates the switch between latent and replicate forms of EBV infection. The aim of this study was to analyze the feasibility of BZLF1 mRNA detection in saliva as a marker for active replication of the virus. Various specimens (saliva, plasma, PBMC) from 17 patients with EBV-induced infectious mononucleosis (IM) and 4 control patients were examined for expression of viral BZLF1 mRNA by means of real-time PCR. BZLF1 expression was correlated to the amount of viral DNA in either compartment. Digestion of plasma and saliva samples with DNase I allowed distinguishing between encapsidated and naked viral DNA. BZLF1 transcripts were found in all different types of specimens in varying frequencies. BZLF1 expression in saliva, PBMC, and plasma correlated with viral load in each compartment. Interestingly, those patients with detectable BZLF1 expression in saliva had a more severe course of infection with longer duration of hospitalization. In conclusion, this study demonstrates the feasibility of BZLF1 mRNA detection in saliva specimens during replicative EBV infection. Its significance for the diagnosis of reactivated EBV infection, particularly under immunosuppression, has to be elucidated in further studies. Copyright © 2017 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. Susceptibility of anthocyanins to ex vivo degradation in human saliva

    Science.gov (United States)

    Kamonpatana, Kom; Giusti, M. Mónica; Chitchumroonchokchai, Chureeporn; MorenoCruz, Maria; Riedl, Ken M.; Kumar, Purnima; Failla, Mark L.

    2013-01-01

    Some fruits and their anthocyanin-rich extracts have been reported to exhibit chemopreventive activity in the oral cavity. Insights regarding oral metabolism of anthocyanins remain limited. Anthocyanin-rich extracts from blueberry, chokeberry, black raspberry, red grape, and strawberry were incubated ex vivo with human saliva from 14 healthy subjects. All anthocyanins were partially degraded in saliva. Degradation of chokeberry anthocyanins in saliva was temperature dependent and decreased by heating saliva to 80 °C and after removal of cells. Glycosides of delphinidin and petunidin were more susceptible to degradation than those of cyanidin, pelargonidin, peonidin and malvidin in both intact and artificial saliva. Stability of di- and tri-saccharide conjugates of anthocyanidins slightly, but significantly, exceeded that of monosaccharide compounds. Ex vivo degradation of anthocyanins in saliva was significantly decreased after oral rinsing with antibacterial chlorhexidine. These results suggest that anthocyanin degradation in the mouth is structure-dependent and largely mediated by oral microbiota. PMID:22868153

  18. Immunoreactive pattern of Staphylococcus epidermidis biofilm against human whole saliva.

    Science.gov (United States)

    Carvalhais, Virginia; Amado, Francisco; Cerveira, Frederico; Ferreira, Rita; Vilanova, Manuel; Cerca, Nuno; Vitorino, Rui

    2015-05-01

    Saliva is essential to interact with microorganisms in the oral cavity. Therefore, the interest in saliva antimicrobial properties is on the rise. Here, we used an immunoproteomic approach, based on protein separation of Staphylococcus epidermidis biofilms by 2DE, followed by Western-blotting, to compare human serum and saliva reactivity profile. A total of 17 proteins were identified by MALDI-TOF/TOF. Serum and saliva presented a distinct pattern of immunoreactive proteins. Our results suggest that saliva seems to have higher propensity to react against S. epidermidis proteins with oxidoreductase activity and proteins involved with L-serine metabolic processes. We show that saliva was a powerful tool for the identification of potential S. epidermidis biofilms proteins. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Cloning of a salivary gland metalloprotease and characterization of gelatinase and fibrin(ogen)lytic activities in the saliva of the Lyme Disease tick vector Ixodes scapularis

    Science.gov (United States)

    Francischetti, Ivo M. B.; Mather, Thomas N.; Ribeiro, José M. C.

    2010-01-01

    The full-length sequence of tick salivary gland cDNA coding for a protein similar to metalloproteases (MP) of the reprolysin family is reported. The Ixodes scapularis MP is a 488 aminoacid (aa) protein containing pre- and pro-enzyme domains, the zinc-binding motif HExxHxxGxxH common to metalloproteases and a cysteine-rich region. In addition, the predicted amino-terminal sequences of I. scapularis MPs were found by Edman degradation of PVDF-transferred SDS/PAGE-separated tick saliva proteins, indicating these putative enzymes are secreted. Furthermore, saliva has a metal-dependent proteolytic activity towards gelatin, fibrin(ogen) and fibronectin, but not collagen or laminin. Accordingly, I. scapularis saliva has a rather specific metalloprotease similar to the hemorrhagic proteases of snake venoms. This is the first description of such activity in tick saliva and its role in tick feeding and Borrelia transmission are discussed. PMID:12767911

  20. Use of saliva as a diagnostic fluid in dentistry

    OpenAIRE

    Todorović Tatjana; Dožić Ivan; Pavlica Dušan; Marković Dejan; Ivanović Mirjana; Brajović Gavrilo; Stefanović Gordana; Mirković Silvija; Anđelski Biljana

    2005-01-01

    Saliva is a secretion of the salivary and mucous glands and is of major importance in the maintainance of oral health. Over the last few decades, saliva has been evaluated as a diagnostic fluid in medicine for determining systemic disease markers as well as for monitoring numerous drugs, narcotics, and hormones. The biochemical analysis of saliva is particularly important in dentistry. The estimation of the risk of appearance and diagnosis of disease, monitoring of disease progression, evalua...

  1. The influence of tooth brushing time over saliva buffering capacity

    OpenAIRE

    Sri Mulyanti; Hetty Anggrawati

    2014-01-01

    Saliva gives a considerable influence against the growth of dental caries as a natural defense against caries. the things very important about saliva are its flow rate and buffering capacity. the decrease in saliva flow rate might cause food retention that furthermore would turn into dental plaques, meanwhile it’s buffering capacity will play a considerable role in maintaining the saliva’s pH and remineralization process of the teeth. One of the mechanisms which are considered to be effective...

  2. Susceptibility of anthocyanins to ex vivo degradation in human saliva

    OpenAIRE

    Kamonpatana, Kom; Giusti, M. Mónica; Chitchumroonchokchai, Chureeporn; MorenoCruz, Maria; Riedl, Ken M.; Kumar, Purnima; Failla, Mark L.

    2012-01-01

    Some fruits and their anthocyanin-rich extracts have been reported to exhibit chemopreventive activity in the oral cavity. Insights regarding oral metabolism of anthocyanins remain limited. Anthocyanin-rich extracts from blueberry, chokeberry, black raspberry, red grape, and strawberry were incubated ex vivo with human saliva from 14 healthy subjects. All anthocyanins were partially degraded in saliva. Degradation of chokeberry anthocyanins in saliva was temperature dependent and decreased by...

  3. Pengaruh Larutan Kumur Ekstrak Siwak (Salvadora Persica) Terhadap Ph Saliva

    OpenAIRE

    Kusumasari, Nila; Santoso, Oedijani

    2012-01-01

    Latar belakang : pH saliva merupakan salah satu komponen yang memberikan kontribusi terhadap pH mulut. Bakteri patogen dalam rongga mulut memfermentasi gula menjadi asam laktat yang akan menurunkan keasaman mulutsehingga menyebabkan demineralisasi email gigi. Untuk mencegah penurunan pH saliva dapat dilakukan secara kimiawi. Pada penelitian ini digunakan larutan ekstrak siwak (Salvadora persica) sebagai obat kumur karena terdapat fitokemikal yang mampu mencegah penurunan pH saliva dengan cara...

  4. [Use of saliva as a diagnostic fluid in dentistry].

    Science.gov (United States)

    Todorović, Tatjana; Dozić, Ivan; Pavlica, Dusan; Marković, Dejan; Brajović, Gavrilo; Ivanović, Mirjana; Stevanović, Gordana; Mirković, Silvija; Andjelski, Biljana

    2005-01-01

    Saliva is a secretion of the salivary and mucous glands and is of major importance in the maintainance of oral health. Over the last few decades, saliva has been evaluated as a diagnostic fluid in medicine for determining systemic disease markers as well as for monitoring numerous drugs, narcotics, and hormones. The biochemical analysis of saliva is particularly important in dentistry. The estimation of the risk of appearance and diagnosis of disease, monitoring of disease progression, evaluation of therapy efficacy for caries, periodontitis, premalignant and malignant oral lesions, as well as infectious diseases of the oral cavity, can be assessed by analysing different constituents of saliva. Individuals at risk of caries can be identified using tests that determine saliva flow rate, saliva buffer capacity, and colonisation of the oral cavity by cariogenic bacteria. Today, these rapid and simple diagnostic tests are used routinely in caries risk determination. The study and use of saliva-based diagnostics have increased over the last few decades. Clinical testing of saliva shows much promise. However, there is a need for much additional research in this area, before the true clinical value of saliva as a diagnostic fluid in dentistry can be determined.

  5. Molecular alterations of parotid saliva in infantile chronic recurrent parotitis.

    Science.gov (United States)

    Morales-Bozo, Irene; Urzúa-Orellana, Blanca; Landaeta, Mirtha; Montalbán, Raúl; Torres, Jimena; Pinochet, Alvaro; Valverde, Gustavo; Muñoz-Martínez, Andrea

    2007-02-01

    Infantile chronic recurrent parotitis (ICRP) is an insidious disease whose etiopathogenesis remains an enigma. Alterations in the physical appearance of parotid saliva from ICRP patients have been frequently reported. However, sialochemical studies in regard to ICRP are very rare. The aim of this study was to determine whether saliva of ICRP patients presents major physicochemical and biochemical alterations compared with saliva from paired healthy controls. Parotid, whole, and submandibular/sublingual saliva was collected at an asymptomatic stage from 33 ICRP patients (5-16 y old, both sexes) and from 33 sex- and age-matched healthy controls. Saliva was analyzed for protein concentration, mode of protein diffusion on cellulose membranes, unidimensional sodium dodecylsulfate (SDS)-polyacrylamide gel electrophoresis protein profiles and zymographic profiles of metalloproteinase 2 (MMP-2) and metalloproteinase 9 (MMP-9). Parotid saliva of ICRP patients showed an increased protein concentration, altered mode of protein diffusion, a higher frequency of polypeptide bands of 43, 37, 33, 29, 26, 16, and 10 kD, higher asymmetry in the polypeptide profiles of both contralateral parotid saliva, and an increase in the frequency of MMP-2 and MMP-9. Parotid saliva of patients with ICRP is molecularly altered with respect to normal saliva. Some of the molecular differences could be related to the etiopathogenesis of the disease.

  6. Use of saliva as a diagnostic fluid in dentistry

    Directory of Open Access Journals (Sweden)

    Todorović Tatjana

    2005-01-01

    Full Text Available Saliva is a secretion of the salivary and mucous glands and is of major importance in the maintainance of oral health. Over the last few decades, saliva has been evaluated as a diagnostic fluid in medicine for determining systemic disease markers as well as for monitoring numerous drugs, narcotics, and hormones. The biochemical analysis of saliva is particularly important in dentistry. The estimation of the risk of appearance and diagnosis of disease, monitoring of disease progression, evaluation of therapy efficacy for caries, periodontitis, premalignant and malignant oral lesions, as well as infectious diseases of the oral cavity, can be assessed by analyzing different constituent: of saliva, individuals at risk of caries can be identified using test: that determine saliva flow rate, saliva buffer capacity, and colonization of the oral cavity by cariogenic bacteria. Today, these rapid and simple diagnostic tests are used routinely in caries risk determination. The study and use of saliva-based diagnostics have increased over the last few decades. Clinical testing of saliva shows much promise. However, there is a need for much additional research in this area, before the true clinical value of saliva as a diagnostic fluid in dentistry can be determined.

  7. Identification of canine saliva using mRNA-based assay.

    Science.gov (United States)

    Nakanishi, Hiroaki; Ohmori, Takeshi; Hara, Masaaki; Yoneyama, Katsumi; Takada, Aya; Saito, Kazuyuki

    2017-01-01

    A dog saliva analysis in addition to a bite-mark analysis may be important for evidence when a crime involves a dog bite. In this study, the utility of detecting canine saliva-specific mRNAs to identify canine saliva was evaluated. Canine saliva swabs (n = 20), urine swabs (n = 20), body surface swabs (n = 20), whole blood samples (n = 10), human saliva (n = 20), human skin surface swabs (n = 20), and human whole blood (n = 20) were tested. The saliva-specific genes encoding statherin (STATH), carbonic anhydrase VI (CA-VI), and dog allergens (Canf1 and Canf2) were analyzed as candidate genes. Moreover, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as confirmation of canine mRNA extraction. STATH, CA-VI, Canf1, Canf2, and GAPDH mRNAs were detected in 19/20, 1/20, 11/20, 4/20, and 20/20 saliva samples, respectively. The STATH, CA-VI, Canf1, Canf2, and GAPDH mRNAs did not exhibit cross-reactivity with samples of human origin. This mRNA-based assay was also able to detect canine saliva in mock forensic samples. The results of this study indicated that the detection of STATH mRNA is useful for the identification of canine saliva, and GAPDH is a suitable marker for canine mRNA extraction.

  8. Automation of DNA and miRNA co-extraction for miRNA-based identification of human body fluids and tissues.

    Science.gov (United States)

    Kulstein, Galina; Marienfeld, Ralf; Miltner, Erich; Wiegand, Peter

    2016-10-01

    In the last years, microRNA (miRNA) analysis came into focus in the field of forensic genetics. Yet, no standardized and recommendable protocols for co-isolation of miRNA and DNA from forensic relevant samples have been developed so far. Hence, this study evaluated the performance of an automated Maxwell® 16 System-based strategy (Promega) for co-extraction of DNA and miRNA from forensically relevant (blood and saliva) samples compared to (semi-)manual extraction methods. Three procedures were compared on the basis of recovered quantity of DNA and miRNA (as determined by real-time PCR and Bioanalyzer), miRNA profiling (shown by Cq values and extraction efficiency), STR profiles, duration, contamination risk and handling. All in all, the results highlight that the automated co-extraction procedure yielded the highest miRNA and DNA amounts from saliva and blood samples compared to both (semi-)manual protocols. Also, for aged and genuine samples of forensically relevant traces the miRNA and DNA yields were sufficient for subsequent downstream analysis. Furthermore, the strategy allows miRNA extraction only in cases where it is relevant to obtain additional information about the sample type. Besides, this system enables flexible sample throughput and labor-saving sample processing with reduced risk of cross-contamination. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. The validation of forensic DNA extraction systems to utilize soil contaminated biological evidence.

    Science.gov (United States)

    Kasu, Mohaimin; Shires, Karen

    2015-07-01

    The production of full DNA profiles from biological evidence found in soil has a high failure rate due largely to the inhibitory substance humic acid (HA). Abundant in various natural soils, HA co-extracts with DNA during extraction and inhibits DNA profiling by binding to the molecular components of the genotyping assay. To successfully utilize traces of soil contaminated evidence, such as that found at many murder and rape crime scenes in South Africa, a reliable HA removal extraction system would often be selected based on previous validation studies. However, for many standard forensic DNA extraction systems, peer-reviewed publications detailing the efficacy on soil evidence is either lacking or is incomplete. Consequently, these sample types are often not collected or fail to yield suitable DNA material due to the use of unsuitable methodology. The aim of this study was to validate the common forensic DNA collection and extraction systems used in South Africa, namely DNA IQ, FTA elute and Nucleosave for processing blood and saliva contaminated with HA. A forensic appropriate volume of biological evidence was spiked with HA (0, 0.5, 1.5 and 2.5 mg/ml) and processed through each extraction protocol for the evaluation of HA removal using QPCR and STR-genotyping. The DNA IQ magnetic bead system effectively removed HA from highly contaminated blood and saliva, and generated consistently acceptable STR profiles from both artificially spiked samples and crude soil samples. This system is highly recommended for use on soil-contaminated evidence over the cellulose card-based systems currently being preferentially used for DNA sample collection. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  10. Non-homologous end-joining protein expression screen from radiosensitive cancer patients yields a novel DNA double strand break repair phenotype.

    Science.gov (United States)

    McKay, Michael J; Goh, Su Kak; McKay, Jeremy N; Chao, Michael; McKay, Timothy M

    2017-03-01

    Clinical radiosensitivity is a significant impediment to tumour control and cure, in that it restricts the total doses which can safely be delivered to the whole radiotherapy population, within the tissue tolerance of potentially radiosensitive (RS) individuals. Understanding its causes could lead to personalization of radiotherapy. We screened tissues from a unique bank of RS cancer patients for expression defects in major DNA double-strand break repair proteins, using Western blot analysis and subsequently reverse-transcriptase polymerase chain reaction and pulsed-field gel electrophoresis. We hypothesized that abnormalities in expression of these proteins may explain the radiosensitivity of some of our cancer patients. The cells from one patient showed a reproducibly consistent expression reduction in two complex-forming DNA double-strand break repair protein components (DNA Ligase IV and XRCC4). We also showed a corresponding reduction in both gene products at the mRNA level. Additionally, the mRNA inducibility by ionizing radiation was increased for one of the proteins in the patient's cells. We confirmed the likely functional significance of the non-homologous end-joining (NHEJ) expression abnormalities with a DNA double strand break (DNA DSB) repair assay. We have identified a novel biological phenotype linked to clinical radiosensitivity. This is important in that very few molecular defects are known in human radiotherapy subjects. Such knowledge may contribute to the understanding of radiation response mechanisms in cancer patients and to personalization of radiotherapy.

  11. Saliva: a potential media for disease diagnostics and monitoring.

    Science.gov (United States)

    Liu, Jingyi; Duan, Yixiang

    2012-07-01

    Within the past 10 years, the use of saliva as a diagnostic tool has gained considerable attention and become a well-accepted method. As a diagnostic fluid, saliva offers superiority over serum due to both a noninvasive collection method by specially trained persons and a cost-effective approach for screening of large populations. Collection of saliva offers a reduced risk of infection compared to the collection of serum. Moreover, obtaining saliva samples from infant, disabled or anxious patients, is much easier than obtaining other samples. There is a lot of useful components-changing information in saliva when a person is in sick. Therefore, we define these changing components as "biomarkers". The utilization of biomarkers as early predictors for clinical disease not only contributes to the effective prevention and treatment of diseases, but also enhances the assessment of potential health risks. In this article, we have reviewed the properties of saliva, the salivary analysis method for biomarker discovery, and the diagnostic potentials of salivary biomarkers in monitoring and detecting periodontal disease, Oral and Breast cancers, and Sjögren's syndrome. We also discussed some barriers of applications of saliva as a diagnostic media as well as recent improvements. We also prospected the future processing directions of using biomarkers in disease diagnosis and draw a conclusion that saliva is indeed an effective media in various disease monitoring and diagnosis. Copyright © 2012 Elsevier Ltd. All rights reserved.

  12. Saliva and plasma TIMP-1 in patients with colorectal cancer

    DEFF Research Database (Denmark)

    Holten-Andersen, Lars; Christensen, Ib Jarle; Jensen, Siri Beier

    2012-01-01

    Background and aims. A prospective cross-sectional study was designed to test if total levels of TIMP-1 in saliva and plasma correlated with the diagnosis of colorectal cancer (CRC) in a population with symptoms consistent with this disease. Materials and methods. Stimulated whole saliva and bloo...

  13. The role of electrostatics in saliva-induced emulsion flocculation

    NARCIS (Netherlands)

    Silletti, Erika; Vingerhoeds, Monique H.; Norde, Willem; Van Aken, George A.

    Upon consumption food emulsions undergo different processes, including mixing with saliva. It has been shown that whole saliva induces emulsion flocculation [van Aken, G. A., Vingerhoeds, M. H., & de Hoog, E. H. A. (2005). Colloidal behaviour of food emulsions under oral conditions. In E. Dickinson

  14. Evaluation of wetting ability of five new saliva substitutes on ...

    African Journals Online (AJOL)

    Introduction: The aim of this study was to evaluate & compare the wetting ability of five saliva substitutes & distilled water on heat-polymerized acrylic resin. Contact angle of the saliva substitute on denture base can be taken as an indicator of wettability. Good wetting of heat-polymerized acrylic resin is critical for optimum ...

  15. Nonenzymatic antioxidants in saliva of patients with systemic lupus erythematosus.

    Science.gov (United States)

    Moori, M; Ghafoori, H; Sariri, R

    2016-03-01

    Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by autoantibody-directed self-antigens, immune complex formation and immune deregulation, resulting in damage to essentially all the organs. SLE is associated with the increased production of free radicals. Increase in free radicals or impaired antioxidant defense system in SLE causes oxidative stress. Considering that saliva could be a reflection of the state of health, the purpose of this study was to evaluate some antioxidants in the saliva and serum of patients with SLE and compare these with healthy individuals. This could help us in obtaining a possible marker in saliva in the future. During the course of the practical part of the project, 30 patients with SLE and 30 healthy controls were investigated. After centrifugation of un-stimulated saliva and blood samples, they were examined using spectrophotometric methods and the results were analyzed by statistical software. According to the results, concentrations of malondialdehyde, uric acid and total antioxidants were significantly increased but the level of reduced glutathion was reduced significantly in the saliva and serum of SLE patients as compared to controls. It is therefore suggested that antioxidant power is impaired in saliva and serum of SLE patients. As there was a positive correlation between the antioxidant level of saliva and blood serum, the antioxidant status of saliva could be an indicator of serum antioxidants. © The Author(s) 2015.

  16. Discovery of Mosquito Saliva MicroRNAs during CHIKV Infection

    Science.gov (United States)

    Maharaj, Payal D.; Widen, Steven G.; Huang, Jing; Wood, Thomas G.; Thangamani, Saravanan

    2015-01-01

    Mosquito borne pathogens are transmitted to humans via saliva during blood feeding. Mosquito saliva is a complex concoction of many secretory factors that modulate the feeding foci to enhance pathogen infection and establishment. Multiple salivary proteins/factors have been identified/characterized that enhance pathogen infection. Here, we describe, for the first time, the identification of exogenous microRNAs from mosquito saliva. MicroRNAs are short, 18–24 nucleotide, non-coding RNAs that regulate gene expression, and are generally intracellular. However, circulating miRNAs have been described from serum and saliva of humans. Exogenous miRNAs have not been reported from hematophagous arthropod saliva. We sought to identify miRNAs in the mosquito saliva and their role in Chikungunya virus (CHIKV) infection. Next generation sequencing was utilized to identify 103 exogenous miRNAs in mosquito saliva of which 31 miRNAs were previously unidentified and were designated novel. Several miRNAs that we have identified are expressed only in the CHIKV infected mosquitoes. Five of the saliva miRNAs were tested for their potential to regulated CHIKV infection, and our results demonstrate their functional role in the transmission and establishment of infection during blood feeding on the host. PMID:25612225

  17. Cell-derived vesicles exposing coagulant tissue factor in saliva.

    Science.gov (United States)

    Berckmans, René J; Sturk, Auguste; van Tienen, Laurens M; Schaap, Marianne C L; Nieuwland, Rienk

    2011-03-17

    On vascular damage, coagulation is initiated by extravascular tissue factor (TF). Intravascular TF, which is present on circulating cell-derived vesicles, is noncoagulant under physiologic conditions but prothrombotic under pathologic conditions. Human saliva triggers coagulation, but the mechanism and physiologic relevance are unknown. Because saliva is known to contain TF, we hypothesized that this TF may also be associated with cell-derived vesicles to facilitate coagulation when saliva directly contacts blood. The saliva-induced shortening of the clotting time of autologous plasma and whole blood from healthy subjects (n = 10) proved TF-dependent. This TF was associated with various types of cell-derived vesicles, including microparticles and exosomes. The physiologic function was shown by adding saliva to human pericardial wound blood collected from patients undergoing cardiac surgery. Addition of saliva shortened the clotting time from 300 ± 96 to 186 ± 24 seconds (P = .03). Our results show that saliva triggers coagulation, thereby reducing blood loss and the risk of pathogens entering the blood. We postulate that our reflex to lick a wound may be a mechanism to enable TF-exposing vesicles, present in saliva, to aid in the coagulation process and thus protect the organism from entering pathogens. This unique compartmentalization may be highly conserved because also animals lick their wounds.

  18. Saliva viscosity as a potential risk factor for oral malodor.

    Science.gov (United States)

    Ueno, Masayuki; Takeuchi, Susumu; Takehara, Sachiko; Kawaguchi, Yoko

    2014-11-01

    The objective of this study was to assess whether saliva viscosity, measured by a viscometer, was a predictor of oral malodor. The subjects were 617 patients who visited an oral malodor clinic. The organoleptic test (OT) was used for diagnosis of oral malodor. An oral examination assessed the numbers of teeth present and decayed teeth as well as the presence or absence of dentures. Further, periodontal pocket depths (PD), gingival bleeding, dental plaque and tongue coating were investigated. Unstimulated saliva were collected for 5 min. Saliva viscosity was measured with a viscometer. Logistic regression analysis with oral malodor status by OT as a dependent variable was performed. Possible confounders including age, gender, number of teeth present, number of decayed teeth, number of teeth with PD ≥ 4 mm, number of teeth with bleeding on probing, presence or absence of dentures, plaque index, area of tongue coating, saliva flow rate, saliva pH and saliva viscosity were used as independent variables. Saliva viscosity (p = 0.047) along with the number of teeth with PD ≥4 mm (p = 0.001), plaque index (p = 0.037) and area of tongue coating (p viscosity (OR = 1.10) were more likely to have oral malodor compared to those with lower values. The results suggested that high saliva viscosity could be a potential risk factor for oral malodor.

  19. Pharmacokinetics of lamotrigine (Lamictal) in plasma and saliva.

    Science.gov (United States)

    Trnavska, Z; Krejcova, H; Tkaczykovam; Salcmanova, Z; Elis, J

    1991-01-01

    The kinetics of lamotrigine (LTG) disposition in plasma and saliva was evaluated in patients undergoing long-term antiepileptic drug therapy. Blood and saliva samples were collected simultaneously at intervals during the study and the concentration of LTG was measured by HPLC. Concentrations of LTG in saliva were proportional to plasma LTG concentrations, with a significant (p) correlation of r = 0.95 + 0.18. The saliva concentration-time curves of LTG were parallel to those derived for plasma LTG. The kinetics of LTG absorption, elimination and mean residence time were identical in both saliva and plasma estimations. The saliva/plasma ratio, determined from the terminal phase of individual patient LTG concentration vs time curves, was used to predict plasma LTG concentrations from saliva determinations. The binding of LTG to plasma proteins remained constant in patients treated with sodium valproate and/or enzyme-inducing drugs. Thus, LTG determination in saliva represents a noninvasive alternative for therapeutic drug monitoring and may also be employed for studying LTG pharmacokinetics.

  20. Effects of different tastants on parotid saliva flow and composition

    NARCIS (Netherlands)

    Neyraud, E.; Heinzerling, C.I.; Bult, J.H.F.; Mesmin, C.; Dransfield, E.

    2009-01-01

    Saliva from parotid glands plays a role in taste perception. Parotid saliva is also stimulated by tastants. The aim of this work is to investigate the effects of different tastants on the parotid salivary response in six subjects. Five tastants were given in different concentrations in solution and

  1. Effects of Different Tastants on Parotid Saliva Flow and Composition

    NARCIS (Netherlands)

    Neyraud, E.; Heinzerling, C.I.; Bult, J.H.F.; Mesmin, C.; Dransfield, E.

    2009-01-01

    Saliva from parotid glands plays a role in taste perception. Parotid saliva is also stimulated by tastants. The aim of this work is to investigate the effects of different tastants on the parotid salivary response in six subjects. Five tastants were given in different concentrations in solution and

  2. Growth of oral Streptococcus species and Actinomyces viscosus in human saliva.

    OpenAIRE

    de Jong, M H; van der Hoeven, J S; van OS, J H; Olijve, J H

    1984-01-01

    Microorganisms in dental plaque live in constant association with saliva. The role of saliva in the adherence of bacteria to the teeth and the antibacterial properties of saliva have been well investigated; less interest has been shown in the possible role of saliva as a substrate for oral microorganisms. In this study it was shown that saliva can serve as a growth medium for oral Streptococcus spp. and Actinomyces viscosus. The cell production of these organisms on saliva was carbohydrate li...

  3. Current development of saliva/oral fluid-based diagnostics.

    Science.gov (United States)

    Yeh, Chih-Ko; Christodoulides, Nicolaos J; Floriano, Pierre N; Miller, Craig S; Ebersole, Jeffrey L; Weigum, Shannon E; McDevitt, John; Redding, Spencer W

    2010-07-01

    Saliva can be easily obtained in medical and non-medical settings, and contains numerous bio-molecules, including those typically found in serum for disease detection and monitoring. In the past two decades, the achievements of high-throughput approaches afforded by biotechnology and nanotechnology allow for disease-specific salivary biomarker discovery and establishment of rapid, multiplex, and miniaturized analytical assays. These developments have dramatically advanced saliva-based diagnostics. In this review, we discuss the current consensus on development of saliva/oral fluid-based diagnostics and provide a summary of recent research advancements of the Texas-Kentucky Saliva Diagnostics Consortium. In the foreseeable future, current research on saliva based diagnostic methods could revolutionize health care.

  4. Raman spectroscopy of human saliva for acute myocardial infarction detection

    Science.gov (United States)

    Chen, Maowen; Chen, Yuanxiang; Wu, Shanshan; Huang, Wei; Lin, Jinyong; Weng, Guo-Xing; Chen, Rong

    2014-09-01

    Raman spectroscopy is a rapidly non-invasive technique with great potential for biomedical research. The aim of this study was to evaluate the feasibility of using Raman spectroscopy of human saliva for acute myocardial infarction (AMI) detection. Raman spectroscopy measurements were performed on two groups of saliva samples: one group from patients (n=30) with confirmed AMI and the other group from healthy controls (n=31). The diagnostic performance for differentiating AMI saliva from normal saliva was evaluated by multivariate statistical analysis. The combination of principal component analysis (PCA) and linear discriminate analysis (LDA) of the measured Raman spectra separated the spectral features of the two groups into two distinct clusters with little overlaps, rendering the sensitivity of 80.0% and specificity of 80.6%. The results from this exploratory study demonstrated that Raman spectroscopy of human saliva can serve as a potentially clinical tool for rapid AMI detection and screening.

  5. Phenotype and Genotype of Enterococcus faecalis Isolated form Root Canal and Saliva of Primary Endodontic Patients

    Directory of Open Access Journals (Sweden)

    Zaki Mubarak

    2016-04-01

    Full Text Available This study was carried out to investigate the phenotype and genotype of E. faecalis isolated from the root canal and saliva of primary endodontic patients with periapical lesions. Eighteen adult male and female individuals suffering from primary endodontic infection, either had or had not periapical lesions, were involved in this study. Root canal scraping and saliva were collected from each subject and used for bacterial quantitation using a real-time polymerase chain reaction (RT-PCR. Enterococci were isolated using ChromAgar medium and then identified using both biochemical (Gram staining and catalase tests and molecular biology (conventional PCR methods. Gelatinase activity, polysaccharide capsul profile and mRNA ace expression level were determined using microbiological, biochemical and molecular biology approach, respectively.  Genotype of E. faecalis was determined based on nucleotide sequence of ace and gelE genes analyzed using web-based 3730xl DNA Analyze software. The results showed that high proportion of E. faecalis found in both root canal and saliva of is related to the incidence of periapical lessions in the primary endodontic patients. This is contrast to the insignificant relationship found between Cps polymorphism, gelatinase activity, and mRNA ace expression with periapical lesions in the patients, respectively.DOI: 10.14693/jdi.v23i1.960

  6. Microbial profiles in saliva from children with and without caries in mixed dentition.

    Science.gov (United States)

    Luo, A H; Yang, D Q; Xin, B C; Paster, B J; Qin, J

    2012-09-01

    The purpose of this study was to determine the bacterial profiles in saliva of the isolated children for studying caries etiology.   Samples were collected from isolated children from 6 to 8years old including 20 caries-free (dmfs=0) (healthy) and 30 caries-active individuals (dmfs>8) (patients). 16S rRNA genes were amplified by PCR from bacterial DNA of saliva sample and labeled via incorporation of Cy3-dCTP in second nested PCR. After hybridization of labeled amplicons on HOMIM, the microarray slides were scanned and original data acquired from professional software. Collectively, 94 bacterial species or clusters representing six bacterial phyla and 30 genera were detected. A higher bacterial diversity was observed in patients than in healthy samples. Statistical analyses revealed eight species or clusters were detected more frequently in diseased patients than in healthy samples, while six different species were detected more frequently in healthy as compared to diseased patients. The diversity of microbe within saliva derived from isolated population increased in caries-active status, and there are some bacteria in salivary flora can be as candidate biomarkers for caries prognosis in mixed dentition. The imbalances in the resident microflora may be the ultimate mechanism of dental caries. © 2012 John Wiley & Sons A/S.

  7. Human saliva exposure modulates bone cell performance in vitro.

    Science.gov (United States)

    Proksch, Susanne; Steinberg, Thorsten; Keller, Constantin; Wolkewitz, Martin; Wiedmann-Al-Ahmad, Margit; Finkenzeller, Guenter; Hannig, Christian; Hellwig, Elmar; Al-Ahmad, Ali

    2012-02-01

    Various situations encountered by a clinician during the daily routine including surgical periodontitis therapy, dental implant insertion, or tooth extraction involve the contact of saliva with the jaw bone. However, there are only sparse data concerning the influence of saliva on bone cells. Saliva specimens were incorporated within culture medium and administered to murine MC3T3 osteoblasts, of which the morphology (REM), proliferation (EZ4U), and differentiation (qRT-PCR, alkaline phosphatase activity, extracellular matrix calcification) were assessed. Simultaneously, the composition of saliva media was analyzed with respect to the content of lactoferrin, activities of classical salivary enzymes, and the ability to provoke inflammatory cytokine production (enzyme-linked immunosorbent assay) in MC3T3 osteoblasts. The morphology, proliferation, and expression of differentiation-associated genes were seriously handicapped by saliva contact. Saliva-touched cells exhibited less alkaline phosphatase but normal levels of extracellular matrix mineralization. Saliva-containing culture media featured physiological activities of salivary enzymes and considerable amounts of lactoferrin but almost completely lacked salivary alkaline phosphatase and unspecific proteases. Upon saliva incubation, MC3T3 osteoblasts did not release noteworthy levels of interleukin-1 beta or tumor necrosis factor alpha. Although saliva is generally considered to vitalize oral tissues, this study reveals that it harms osteoblast-like cells more due to the presence of salivary enzymes than by triggering of inflammation. This issue is clinically relevant because it broadens the understanding of the bone cell fate within the rather complex cosmos of the oral cavity thereby providing a basis for clinical decision making and treatment guidelines. It seems to be reasonable to restrict the contact period between saliva and bone.

  8. Prevalence of human papillomavirus in saliva of women with HPV genital lesions.

    Science.gov (United States)

    Visalli, Giuseppa; Currò, Monica; Facciolà, Alessio; Riso, Romana; Mondello, Placido; Laganà, Pasqualina; Di Pietro, Angela; Picerno, Isa; Spataro, Pasquale

    2016-01-01

    The human papilloma viruses (HPVs) are DNA viruses associated with benign and malignant lesions of skin and mucous membranes. The HPVs has been implicated as the cause of virtually all cervical cancers worldwide but studies showed that these viruses can cause numerous cancers in several tissues including Oral Squamous Cell Carcinoma (OSCC). At least 90 % of HPV-positive OSCCs are associated with high-risk (or oncogenic) HPV-16 and oral infection confers an approximate 50-fold increase in risk for HPV-positive OSCC. HPV-positive OSCCs are associated with sexual behaviors in contrast to HPV-negative OSCCs that are associated with chronic tobacco and alcohol use. The aim of this study was to estimate the prevalence of HPV-DNA in saliva samples collected from women in which it has been previously established the HPV infection of the cervix with relative genotyping and, then, to study the possible correlation. Saliva samples were collected from 100 women with HPV cervical lesions, aged between 22 and 52 years old, and 25 healthy women with normal cytology (control group), aged between 20 and 49 years old. PCR assay was used to detect HPV DNA. The prevalence of oral HPV infection in saliva samples was 24 % in women with HPV cervical lesions while in the control group was 8 %. It has been demonstrated a strong association between high grade squamous intraepithelial lesion and oral infection due to HPV16 and 18, that are the most frequently detected HPV genotypes. This study shows that patients with genital HPV infection are at risk for oral infection and, consequently, for the development of OSCC.

  9. Rheological behavior of food emulsions mixed with saliva : Effect of oil content, salivary protein content, and saliva type

    NARCIS (Netherlands)

    Silletti, Erika; Vingerhoeds, Monique H.; Van Aken, George A.; Norde, Willem

    In this paper, we studied the effect of saliva on the rheological properties of beta-lactoglobulin- and lysozyme-stabilized emulsions, prepared at pH=6.7 in relation to variation of emulsions- and saliva-related parameters. The effect of oil-volume fraction (2.5% w/w to 10% w/w), salivary protein

  10. Rheological Behavior of Food Emulsions Mixed with Saliva: Effect of Oil Content, Salivary Protein Content, and Saliva Type

    NARCIS (Netherlands)

    Silletti, E.; Vingerhoeds, M.H.; Aken, van G.A.; Norde, W.

    2008-01-01

    In this paper, we studied the effect of saliva on the rheological properties of ß-lactoglobulin- and lysozyme-stabilized emulsions, prepared at pH¿=¿6.7 in relation to variation of emulsions- and saliva-related parameters. The effect of oil¿volume fraction (2.5% w/w to 10% w/w), salivary protein

  11. 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

  12. Raman spectroscopy of saliva as a perspective method for periodontitis diagnostics Raman spectroscopy of saliva

    Science.gov (United States)

    Gonchukov, S.; Sukhinina, A.; Bakhmutov, D.; Minaeva, S.

    2012-01-01

    In view of its potential for biological tissues analyses at a molecular level, Raman spectroscopy in optical range has been the object of biomedical research for the last years. The main aim of this work is the development of Raman spectroscopy for organic content identifying and determination of biomarkers of saliva at a molecular level for periodontitis diagnostics. Four spectral regions were determined: 1155 and 1525 cm-1, 1033 and 1611 cm-1, which can be used as biomarkers of this widespread disease.

  13. Development of a radiation track structure clustering algorithm for the prediction of DNA DSB yields and radiation induced cell death in Eukaryotic cells.

    Science.gov (United States)

    Douglass, Michael; Bezak, Eva; Penfold, Scott

    2015-04-21

    The preliminary framework of a combined radiobiological model is developed and calibrated in the current work. The model simulates the production of individual cells forming a tumour, the spatial distribution of individual ionization events (using Geant4-DNA) and the stochastic biochemical repair of DNA double strand breaks (DSBs) leading to the prediction of survival or death of individual cells. In the current work, we expand upon a previously developed tumour generation and irradiation model to include a stochastic ionization damage clustering and DNA lesion repair model. The Geant4 code enabled the positions of each ionization event in the cells to be simulated and recorded for analysis. An algorithm was developed to cluster the ionization events in each cell into simple and complex double strand breaks. The two lesion kinetic (TLK) model was then adapted to predict DSB repair kinetics and the resultant cell survival curve. The parameters in the cell survival model were then calibrated using experimental cell survival data of V79 cells after low energy proton irradiation. A monolayer of V79 cells was simulated using the tumour generation code developed previously. The cells were then irradiated by protons with mean energies of 0.76 MeV and 1.9 MeV using a customized version of Geant4. By replicating the experimental parameters of a low energy proton irradiation experiment and calibrating the model with two sets of data, the model is now capable of predicting V79 cell survival after low energy (cell survival probability, the cell survival probability is calculated for each cell in the geometric tumour model developed in the current work. This model uses fundamental measurable microscopic quantities such as genome length rather than macroscopic radiobiological quantities such as alpha/beta ratios. This means that the model can be theoretically used under a wide range of conditions with a single set of input parameters once calibrated for a given cell line.

  14. Fusion of antigen to a dendritic cell targeting chemokine combined with adjuvant yields a malaria DNA vaccine with enhanced protective capabilities.

    Science.gov (United States)

    Luo, Kun; Zhang, Hong; Zavala, Fidel; Biragyn, Arya; Espinosa, Diego A; Markham, Richard B

    2014-01-01

    Although sterilizing immunity to malaria can be elicited by irradiated sporozoite vaccination, no clinically practical subunit vaccine has been shown to be capable of preventing the approximately 600,000 annual deaths attributed to this infection. DNA vaccines offer several potential advantages for a disease that primarily affects the developing world, but new approaches are needed to improve the immunogenicity of these vaccines. By using a novel, lipid-based adjuvant, Vaxfectin, to attract immune cells to the immunization site, in combination with an antigen-chemokine DNA construct designed to target antigen to immature dendritic cells, we elicited a humoral immune response that provided sterilizing immunity to malaria challenge in a mouse model system. The chemokine, MIP3αCCL20, did not significantly enhance the cellular infiltrate or levels of cytokine or chemokine expression at the immunization site but acted with Vaxfectin to reduce liver stage malaria infection by orders of magnitude compared to vaccine constructs lacking the chemokine component. The levels of protection achieved were equivalent to those observed with irradiated sporozoites, a candidate vaccine undergoing development for further large scale clinical trial. Only vaccination with the combined regimen of adjuvant and chemokine provided 80-100% protection against the development of bloodstream infection. Treating the immunization process as requiring the independent steps of 1) attracting antigen-presenting cells to the site of immunization and 2) specifically directing vaccine antigen to the immature dendritic cells that initiate the adaptive immune response may provide a rational strategy for the development of a clinically applicable malaria DNA vaccine.

  15. 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.

  16. Determination of ovulation in women using saliva ferning test

    Directory of Open Access Journals (Sweden)

    Riska Mutia Ersyari

    2014-11-01

    Full Text Available Every human being experiences growth and development, starting from childhood to adulthood. Women who have entered puberty will experience monthly menstrual cycle. One phase of the menstrual cycle is ovulation or the fertile phase of a woman. The fertile period is the period in which there is an egg ready to be fertilized by sperm. At the time of fertility, there is an increase in the amount of estrogen and progesterone hormones. Increase in these hormones is also found in saliva. Saliva as a biological fluid in the body can be used as a diagnostic fluid. Woman’s fertile period can be assessed from the saliva. Saliva containing high estrogen hormones can form a ferning picture on saliva dried on object glass. The type of research is the study of literature. A literature study was conducted to discuss the determining of the fertile woman with saliva ferning test. The results of previous studies showed the existence of differences in saliva pictures at the time of the fertile period and the infertile period. Salivary ferning was very clearly seen in the woman’s fertile period.

  17. [Serology of the human immunodeficiency virus in saliva].

    Science.gov (United States)

    Cárcaba, V; Fernández, E; Rodríguez Junquera, M; García Amorín, Z; Alfonso, J; García Alonso, S

    1993-07-03

    The presence of immunoglobulins in saliva has allowed it to be proven that they are specific against certain antigens. Antibodies to the human immunodeficiency virus (HIV) have been observed in saliva. The aim of this study was to evaluate the detection of the same by commercial enzymoinmmunoassay (EIA) and standardize the technique. In 78 intravenous drug user patients the presence of antibodies against HIV in serum and saliva were determined by recombinant EIA (Abbott HIV-1/HIV-2 recombinant EIA). The determinations in saliva were made taking volumes of 10 and 50 microliters. In 43 patients the presence of antibodies against HIV-1 was demonstrated in serum, 42 of which were positive in saliva in the determination with 50 microliters and 16 with 10 microliters. No false positives were reported. With the use of 50 microliters of saliva the test showed a sensitivity of 0.98, specificity of 1, predictive value of a positive result of 1, predictive value of negative result of 0.98 and diagnostic efficacy of 0.99. The determination of antibodies against HIV in saliva in intravenous drug users is a highly sensitive and specific method with the use of volumes of 50 microliters in the tests.

  18. 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.

  19. yield indicators

    African Journals Online (AJOL)

    YIELD INDICATORS. P. NTAWURUHUNGA, P.R. RUBAIHAYOI, J.B.A. WHYTE, A.G.O. DIXONZ and use. osnzu1. International Institute of Tropical Agriculture, East and Southern Africa, Centre, PO. Box 7878, l Kampala ... most important sources of food energy in several ... efficiency in selecting and identifying cassava.

  20. Saliva-Based Screening of High-Risk Human Papillomavirus Strains: Detection in Female Indonesian and Thai Dental Students.

    Science.gov (United States)

    Wimardhani, Yuniardini Septorini; Sasanti, Harum; Wardhany, Indriasti Indah; Sarsito, Afi Savitri; Pradono, Siti Aliyah; Subita, Gus Permana; Soegyanto, Anandina Irmagita; Rahmayanti, Febrina; Chamusri, Nutchapon; Iamaroon, Anak

    2015-01-01

    Currently it is believed that human papillomaviruses (HPV) are associated with the development of some oral/oropharyngeal cancers. It has been suggested that these viruses influence carcinogenesis in both smokers and non-smokers. Data on the prevalence of HPV in healthy adults are thus needed to estimate the risk of oral/oropharyngeal cancer. The aim of this study was to assess the prevalence of oral HPV in healthy female adults in Indonesia and Thailand. Healthy female students from the Faculties of Dentistry of Universitas Indonesia and Chiang Mai University were asked to participate in this pilot study. DNA was extracted from saliva specimens and screened for HPV16 and HPV18 using PCR. The age, marital status and sexual experience of the subjects between the two countries were not significantly different. Eight (4%) and 4 (2%) samples were positive for HPV16 and HPV18, respectively. Fisher's Exact test found a significant difference between HPV16 positivity in subjects who were married and had sexual intercourse but not for HPV18. This study successfully detected presence of HPV16 and HPV18 DNA in a number of saliva samples from female dental school students. Marital status, experience of sexual intercourse and safe sexual practice are related to the possibility of finding HPV DNA finding in saliva. Dentists, physicians and other health care professionals may gain significant value from the findings of this study, which provide an understanding of the nature of HPV infection and its risk to patient health and disease.

  1. Establishing a novel automated magnetic bead-based method for the extraction of DNA from a variety of forensic samples.

    Science.gov (United States)

    Witt, Sebastian; Neumann, Jan; Zierdt, Holger; Gébel, Gabriella; Röscheisen, Christiane

    2012-09-01

    Automated systems have been increasingly utilized for DNA extraction by many forensic laboratories to handle growing numbers of forensic casework samples while minimizing the risk of human errors and assuring high reproducibility. The step towards automation however is not easy: The automated extraction method has to be very versatile to reliably prepare high yields of pure genomic DNA from a broad variety of sample types on different carrier materials. To prevent possible cross-contamination of samples or the loss of DNA, the components of the kit have to be designed in a way that allows for the automated handling of the samples with no manual intervention necessary. DNA extraction using paramagnetic particles coated with a DNA-binding surface is predestined for an automated approach. For this study, we tested different DNA extraction kits using DNA-binding paramagnetic particles with regard to DNA yield and handling by a Freedom EVO(®)150 extraction robot (Tecan) equipped with a Te-MagS magnetic separator. Among others, the extraction kits tested were the ChargeSwitch(®)Forensic DNA Purification Kit (Invitrogen), the PrepFiler™Automated Forensic DNA Extraction Kit (Applied Biosystems) and NucleoMag™96 Trace (Macherey-Nagel). After an extensive test phase, we established a novel magnetic bead extraction method based upon the NucleoMag™ extraction kit (Macherey-Nagel). The new method is readily automatable and produces high yields of DNA from different sample types (blood, saliva, sperm, contact stains) on various substrates (filter paper, swabs, cigarette butts) with no evidence of a loss of magnetic beads or sample cross-contamination. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  2. [Mineral composition of mixed saliva in patients with dental fluorosis].

    Science.gov (United States)

    Krikheli, N I; Karamysheva, E I; Lukina, G I; Dubova, L V

    2017-01-01

    The aim of the study was to assess the mineral composition of mixed saliva in dental fluorosis patients undergoing treatment with microabrasion and bleaching. The study included 60 patients aged 18-35 years with various forms of dental fluorosis. Group 1 included 40 patients in which enamel microabrasion was performed, group 2 - 20 patients with microabrasion and bleaching. Mixed saliva composition was analyzed with Olimpus automatic analyzing device. Dental fluorosis treatment in both groups resulted in saliva mineral composition changed associated with enamel demineralization which proves the necessity for calcium and phosphate containing compositions in these treatment groups.

  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. Pesquisa do vírus herpes simples na saliva de pacientes com paralisia facial periférica de Bell Herpes simplex virus in the saliva of peripheral Bell’s palsy patients

    Directory of Open Access Journals (Sweden)

    Paulo Roberto Lazarini

    2006-02-01

    Full Text Available Os primeiros herpes-vírus a serem descritos foram os tipos 1 e 2, cuja denominação é herpes simplex 1 e 2 ou HSV-1 e HSV-2. Estes vírus possuem características biológicas particulares, tais como a capacidade de causar diferentes tipos de doenças, assim como estabelecer infecções latentes ou persistentes por toda a vida dos hospedeiros e de serem reativados causando lesões que podem se localizar no sítio da infecção primária inicial ou próxima a ele. Postula-se que a reativação deste vírus no gânglio geniculado esteja relacionada com a paralisia de Bell. Nesta situação, os vírus, que estariam latentes neste gânglio, sofreriam reativação e replicação difundindo-se pelo nervo facial e seus ramos, dentre eles o nervo corda do tímpano, que ao estimular a secreção salivar possibilitaria a identificação do DNA viral na saliva dos pacientes. Até recentemente, um grande número de pacientes eram diagnosticados como portadores de uma forma desta paralisia, chamada de idiopática ou de paralisia de Bell. Com o advento da técnica de estudo do DNA viral pelo método da reação da polimerase em cadeia (PCR, diversos autores encontraram DNA do vírus herpes simplex tipo I no líquido cefalorraquidiano, na secreção lacrimal, na saliva e nos gânglios geniculados de pacientes com paralisia de Bell. OBJETIVO: observar a prevalência do vírus herpes simplex tipo I pela técnica de PCR, na saliva de pacientes com PFP de Bell, relacionando-a com a evolução clínica destes casos. METODOLOGIA: Avaliamos 38 pacientes portadores de Paralisia Facial Periférica de Bell, que foram submetidos a anamnese, exame médico geral e otorrinolaringológico e coleta de saliva para detecção do DNA viral pela técnica de PCR. O grupo controle correspondeu a 10 adultos normais. RESULTADOS: Obtivemos positividade para o DNA viral em 11 casos dos 38 avaliados, o que corresponde a 29% da amostra. Este resultado foi estatisticamente significante

  5. The diagnosis of metachromatic leucodystrophy during life: metachromatic lipids in saliva and cerebrospinal fluid sediments and in the parotid glands

    Directory of Open Access Journals (Sweden)

    Horacio M. Canelas

    1964-06-01

    Full Text Available The authors report the study of saliva sediment in 4 cases of juvenile metachromatic leucodystrophy belonging to the same family (and else in a sister of one of these cases presenting the characteristic neurological picture but with no metachromasia demonstrable by the Austin test in urine or by biopsies, in 6 normal relatives of the patients with Scholz disease, in 9 cases of various diseases of the nervous system, and in 10 normal subjects. The presence of metachromatic bodies staining in a pinkish red colour with acid blue toluidine dye was demonstrated in the saliva sediment of the 4 cases of metachromatic leucodystrophy. In 2 of these patients biopsies of the parotid gland, stained with cresyl violet dye, showed the presence of intracellular brownish metachromatic bodies. In these 2 cases the study of cerebrospinal fluid sediment also disclosed the presence of metachromatic bodies. Furthermore, a chromatographic qualitative test for metachromatic lipids yielded positive results in saliva, cerebrospinal fluid, and urine sediments. The conclusion was drawn that the search for metachromatic bodies in cerebrospinal fluid and mainly in saliva sediment may be of help in disclosing or ratifying the diagnosis of metachromatic leucodystrophy during life.

  6. Homocysteine measurement in pig saliva, assay validation and changes after acute stress and experimental inflammation models: A pilot study.

    Science.gov (United States)

    Tecles, F; Escribano, D; Martínez-Miró, S; Cerón, J J

    2017-06-01

    High homocysteine (Hcy) concentration in serum has been associated to stress and inflammation in humans, but this association has not been studied in saliva in any animal species. The purpose of this research was to study salivary Hcy levels in pigs under stressful and inflammatory conditions. A commercially available enzyme-linked immunosorbent assay specific for Hcy determination in pigs was adapted and validated in saliva, yielding reproducible and accurate results. Hcy was measured in paired serum-saliva samples and no correlation was observed between serum and salivary Hcy. Salivary Hcy was measured in two experimental models of stress induction in pigs: restraint with a nasal snare and isolation. Homocysteine concentration and the homocysteine to total protein (Hcy/TP) ratio significantly increased 15min after restraining and decreased after some days of isolation. Significant correlation was observed between Hcy and chromogranin A. After an experimentally induced inflammation by subcutaneous turpentine injection, salivary Hcy increased only 3h after turpentine administration; however, the Hcy/TP ratio did not show any change. No correlation was found between salivary Hcy and serum C-reactive protein. In conclusion, salivary Hcy concentration increased when pigs were restrained with a nasal snare or stressed by isolation, probably reflecting an increase in the sympathetic activity. On the other hand, Hcy increased after an experimental inflammation induced by turpentine administration but in this case probably reflects an increase in total protein production in saliva. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. La saliva y sistemas adhesivos alternativos para protesis total

    National Research Council Canada - National Science Library

    Madrid Troconis, Cristhian Camilo; Mendez Silva, Javier Enrique; Tirado Amador, Lesbia Rosa

    2013-01-01

    ... y disminucion del flujo salival. Esta ultima circunstancia es de principal preocupacion, ya que la saliva tiene un papel importante en la retencion de las protesis como "adhesivo natural", por lo que durante anos se han propuesto...

  8. Saliva in relation to dental erosion before and after radiotherapy

    DEFF Research Database (Denmark)

    Jensdottir, Thorbjorg; von Buchwald, Christian; Nauntofte, Birgitte

    2013-01-01

    Abstract Objective. Low saliva flow and abnormal saliva composition are common conditions after radiotherapy for oral cavity and pharyngeal cancer. Both conditions increase the susceptibility to dental caries and erosion, which may be further accelerated by changes in food preferences. The aim...... of this study was to determine changes in saliva flow and susceptibility to erosive challenges in pharyngeal cancer patients before and after radiotherapy to the head and neck. Materials and methods: The erosive potential of sucking acidic candies with and without calcium was determined in nine patients (50...... rates ∼ 17-fold before as well as after radiotherapy (p radiotherapy. Also, saliva became more under-saturated with respect to HAp during (p

  9. Use of saliva as a diagnostic fluid in dentistry

    National Research Council Canada - National Science Library

    Todorovic, Tatjana; Dozic, Ivan; Pavlica, Dusan; Markovic, Dejan; Ivanovic, Mirjana; Brajovic, Gavrilo; Stefanovic, Gordana; Mirkovic, Silvija; Andjelski, Biljana

    2005-01-01

    .... Over the last few decades, saliva has been evaluated as a diagnostic fluid in medicine for determining systemic disease markers as well as for monitoring numerous drugs, narcotics, and hormones...

  10. Microfluidic Immunoassays as Rapid Saliva-Based Clinical Diagnostics

    National Research Council Canada - National Science Library

    Amy E. Herr; Anson V. Hatch; Daniel J. Throckmorton; Huu M. Tran; James S. Brennan; William V. Giannobile; Anup K. Singh

    2007-01-01

    .... Here we report on a clinical POC diagnostic that enables rapid quantitation of an oral disease biomarker in human saliva by using a monolithic disposable cartridge designed to operate in a compact analytical instrument...

  11. Does saliva composition affect the formation of sialolithiasis?

    DEFF Research Database (Denmark)

    Schrøder, Stine; Homøe, preben; Wagner, Niels

    2017-01-01

    with sialolithiasis and 40 matched healthy controls. Patients were examined before and after sialolithiasis surgery; controls were examined once. Flow rate and the inorganic saliva composition in unstimulated whole saliva were assessed. Patients’ salivary flow prior to surgery was significantly lower compared......Saliva composition may affect sialolithiasis formation; thus, this study compared the salivary inorganic composition of sialolithiasis patients with that of healthy controls, and determined whether salivary inorganic composition changes after sialolithiasis surgery. The study included 40 patients...... to that of healthy controls, but equalised after surgery. Prior to surgery, patients’ saliva exhibited higher concentrations of calcium, magnesium, phosphorous compared to that of healthy controls. The concentration of most ions remained high after sialolithiasis surgery. Sialolithiasis patients had increased...

  12. How much pilocarpine contaminates pilocarpine-induced tick saliva?

    Science.gov (United States)

    Ribeiro, J M C; Zeidner, N S; Ledin, K; Dolan, M C; Mather, T N

    2004-03-01

    Pilocarpine is often applied or injected into ticks to induce salivation, and the resulting saliva used to test for various pharmacological, biochemical and immunological activities. To measure the amount of pilocarpine in pilocarpine-induced tick saliva, an HPLC-MS/MS method, based on capillary strong cation exchange chromatography online with an ion trap mass spectrometer, was used to measure pilocarpine in the pg to ng range. Results indicate large concentrations of pilocarpine in Ixodes scapularis Say and Amblyomma americanum (Linnaeus) (Acari: Ixodidae) saliva, ranging from 3 to 50 mm. Due to the known effects of pilocarpine on smooth muscle and immune cells, appropriate controls are proposed and discussed for proper interpretation of results using this saliva preparation.

  13. Artificial saliva effect on toxic substances release from acrylic resins

    OpenAIRE

    Kostić Milena; Krunić Nebojša; Najman Stevo; Nikolić Ljubiša; Nikolić Vesna; Rajković Jelena; Petrović Milica; Igić Marko; Ignjatović Aleksandra

    2015-01-01

    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 a...

  14. Current Development of Saliva/Oral fluid-based Diagnostics

    OpenAIRE

    Yeh, Chih-Ko; Christodoulides, Nicolaos J.; Floriano, Pierre N.; Miller, Craig S.; Ebersole, Jeffrey L.; Weigum, Shannon E.; McDevitt, John; Redding, Spencer W.

    2010-01-01

    Saliva can be easily obtained in medical and non-medical settings, and contains numerous bio-molecules, including those typically found in serum for disease detection and monitoring. In the past two decades, the achievements of high-throughput approaches afforded by biotechnology and nanotechnology allow for disease-specific salivary biomarker discovery and establishment of rapid, multiplex, and miniaturized analytical assays. These developments have dramatically advanced saliva-based diagnos...

  15. Oral glucose tolerance test in unstimulated saliva of healthy individuals

    OpenAIRE

    Mohammad-Hossein Mirzaii-Dizgah; Iraj Mirzaii-Dizgah; Mohammad-Reza Mirzaii-Dizgah

    2016-01-01

    Objective: The aim of this study was to investigate oral glucose tolerance test (OGTT) in unstimulated whole saliva as a diagnostic specimen in clinical practice for detection of diabetes mellitus (DM). Materials and Methods: An interventional study was carried out in 30 apparently healthy individuals aged 24–59 years. Serum and saliva samples were obtained in fasting, 1 h and 2 h after glucose intake (75 g). Glucose concentration was determined by enzymatic colorimetric glucose oxidase-prost...

  16. Detection of chikungunya virus in saliva and urine.

    Science.gov (United States)

    Musso, Didier; Teissier, Anita; Rouault, Eline; Teururai, Sylviane; de Pina, Jean-Jacques; Nhan, Tu-Xuan

    2016-06-16

    Saliva and urine have been used for arthropod-borne viruses molecular detection but not yet for chikungunya virus (CHIKV). We investigated the use of saliva and urine for molecular detection of CHIKV during the French Polynesian outbreak. During the French Polynesian chikungunya outbreak (2014-2015), we collected the same day blood and saliva samples from 60 patients with probable chikungunya (47 during the 1st week post symptoms onset and 13 after), urine was available for 39 of them. All samples were tested using a CHIKV reverse-transcription PCR. Forty eight patients had confirmed chikungunya. For confirmed chikungunya presenting during the 1st week post symptoms onset, CHIKV RNA was detected from 86.1 % (31/36) of blood, 58.3 % (21/36) of saliva and 8.3 % (2/24) of urine. Detection rate of CHIKV RNA was significantly higher in blood compared to saliva. For confirmed chikungunya presenting after the 1st week post symptoms onset, CHIKV RNA was detected from 8.3 % (1/12) of blood, 8.3 % (1/12) of saliva and 0 % (0/8) of urine. In contrast to Zika virus (ZIKV), saliva did not increased the detection rate of CHIKV RNA during the 1st week post symptoms onset. In contrast to ZIKV, dengue virus and West Nile virus, urine did not enlarged the window of detection of CHIKV RNA after the 1st week post symptoms onset. Saliva can be used for molecular detection of CHIKV during the 1st week post symptoms onset only if blood is impossible to collect but with a lower sensitivity compared to blood.

  17. Protein Biomarkers of Periodontitis in Saliva

    Science.gov (United States)

    Taylor, John J.

    2014-01-01

    Periodontitis is a chronic inflammatory condition of the tissues that surround and support the teeth and is initiated by inappropriate and excessive immune responses to bacteria in subgingival dental plaque leading to loss of the integrity of the periodontium, compromised tooth function, and eventually tooth loss. Periodontitis is an economically important disease as it is time-consuming and expensive to treat. Periodontitis has a worldwide prevalence of 5–15% and the prevalence of severe disease in western populations has increased in recent decades. Furthermore, periodontitis is more common in smokers, in obesity, in people with diabetes, and in heart disease patients although the pathogenic processes underpinning these links are, as yet, poorly understood. Diagnosis and monitoring of periodontitis rely on traditional clinical examinations which are inadequate to predict patient susceptibility, disease activity, and response to treatment. Studies of the immunopathogenesis of periodontitis and analysis of mediators in saliva have allowed the identification of many potentially useful biomarkers. Convenient measurement of these biomarkers using chairside analytical devices could form the basis for diagnostic tests which will aid the clinician and the patient in periodontitis management; this review will summarise this field and will identify the experimental, technical, and clinical issues that remain to be addressed before such tests can be implemented. PMID:24944840

  18. 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.

  19. The Saliva Exposome for Monitoring of Individuals' Health Trajectories.

    Science.gov (United States)

    Bessonneau, Vincent; Pawliszyn, Janusz; Rappaport, Stephen M

    2017-07-20

    There is increasing evidence that environmental, rather than genetic, factors are the major causes of most chronic diseases. By measuring entire classes of chemicals in archived biospecimens, exposome-wide association studies (EWAS) are being conducted to investigate associations between a myriad of exposures received during life and chronic diseases. Because the intraindividual variability in biomarker levels, arising from changes in environmental exposures from conception onwards, leads to attenuation of exposure-disease associations, we posit that saliva can be collected repeatedly in longitudinal studies to reduce exposure-measurement errors in EWAS. From the literature and an open-source saliva-metabolome database, we obtained concentrations of 1,233 chemicals that had been detected in saliva. We connected salivary metabolites with human metabolic pathways and PubMed Medical Subject Heading (MeSH) terms, and performed pathway enrichment and pathway topology analyses. One hundred ninety-six salivary metabolites were mapped into 49 metabolic pathways and connected with human metabolic diseases, central nervous system diseases, and neoplasms. We found that the saliva exposome represents at least 14 metabolic pathways, including amino acid metabolism, TCA cycle, gluconeogenesis, glutathione metabolism, pantothenate and CoA biosynthesis, and butanoate metabolism. Saliva contains molecular information worthy of interrogation via EWAS. The simplicity of specimen collection suggests that saliva offers a practical alternative to blood for measurements that can be used to characterize individual exposomes. https://doi.org/10.1289/EHP1011.

  20. Pharmacokinetic Modeling of Intranasal Scopolamine in Plasma Saliva and Urine

    Science.gov (United States)

    Wu, L.; Tam, V. H.; Chow, D. S. L.; Putcha, L.

    2015-01-01

    An intranasal gel dosage formulation of scopolamine (INSCOP) was developed for the treatment of Space Motion Sickness (SMS). The bioavailability and pharmacokinetics (PK) were evaluated under IND (Investigational New Drug) guidelines. The aim of the project was to develop a PK model that can predict the relationships among plasma, saliva and urinary scopolamine concentrations using data collected from the IND clinical trial protocol with INSCOP. Twelve healthy human subjects were administered at three dose levels (0.1, 0.2 and 0.4 mg) of INSCOP. Serial blood, saliva and urine samples were collected between 5 min to 24 h after dosing and scopolamine concentrations were measured by using a validated LC-MS-MS assay. PK compartmental models, using actual dosing and sampling time, were established using Phoenix (version 1.2). Model selection was based on a likelihood ratio test on the difference of criteria (-2LL (i.e. log-likelihood ratio test)) and comparison of the quality of fit plots. The results: Predictable correlations among scopolamine concentrations in compartments of plasma, saliva and urine were established, and for the first time the model satisfactorily predicted the population and individual PK of INSCOP in plasma, saliva and urine. The model can be utilized to predict the INSCOP plasma concentration by saliva and urine data, and it will be useful for monitoring the PK of scopolamine in space and other remote environments using non-invasive sampling of saliva and/or urine.

  1. 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

  2. Total antioxidant capacity of saliva in children with HIV.

    Science.gov (United States)

    Padmanabhan, Vivek; Rai, Kavita; Hegde, Amitha M; Shetty, Sucheta

    2010-01-01

    Several recent reports have indicated high levels of reactive oxygen species, causing oxidative stress, in the pathogenesis of HIV infection. Oxidative stress may lead to enhanced HIV replication in infected cells and may also aggravate the immunodeficiency by reduction of cellular immunity and possibly by increased programmed cell death of lymphocytes. Saliva can constitute a first line of defense against free radical mediated oxidative stress. The use of saliva as a diagnostic fluid has become somewhat of a translational research success story. Technologies are now available enabling saliva to be used to diagnose disease and predict disease progression. The antioxidant capacity of saliva was investigated in 68 children who were divided into two groups. 34 children who were investigated were diagnosed as having HIV infection and the other group consisted of children who reported to the department and served as healthy controls. Total antioxidant capacity of saliva was evaluated by spectrophotometric assay. The results indicated that the total antioxidant capacity (TAC) of saliva decreased in children with HIV infection. TAC was seen to increase with the age of the children.

  3. Susceptibility of anthocyanins to ex vivo degradation in human saliva.

    Science.gov (United States)

    Kamonpatana, Kom; Giusti, M Mónica; Chitchumroonchokchai, Chureeporn; MorenoCruz, Maria; Riedl, Ken M; Kumar, Purnima; Failla, Mark L

    2012-11-15

    Some fruits and their anthocyanin-rich extracts have been reported to exhibit chemopreventive activity in the oral cavity. Insights regarding oral metabolism of anthocyanins remain limited. Anthocyanin-rich extracts from blueberry, chokeberry, black raspberry, red grape, and strawberry were incubated ex vivo with human saliva from 14 healthy subjects. All anthocyanins were partially degraded in saliva. Degradation of chokeberry anthocyanins in saliva was temperature dependent and decreased by heating saliva to 80 °C and after removal of cells. Glycosides of delphinidin and petunidin were more susceptible to degradation than those of cyanidin, pelargonidin, peonidin and malvidin in both intact and artificial saliva. Stability of di- and tri-saccharide conjugates of anthocyanidins slightly, but significantly, exceeded that of monosaccharide compounds. Ex vivo degradation of anthocyanins in saliva was significantly decreased after oral rinsing with antibacterial chlorhexidine. These results suggest that anthocyanin degradation in the mouth is structure-dependent and largely mediated by oral microbiota. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. Comparative analysis of CRT Buffer, GC saliva check buffer tests and laboratory titration to evaluate saliva buffering capacity.

    Science.gov (United States)

    Maldupa, Ilze; Brinkmane, Anda; Mihailova, Anna

    2011-01-01

    OBJECTIVE. The purpose of this study is to evaluate the ability of two commercial strip tests and laboratory titration to detect saliva buffer capacity. MATERIALS AND METHODS. Sixty-four patients were examined. Stimulated saliva was collected and buffer capacity was determined with two different chair-side strip tests in addition to immediate transportation to the laboratory to check the buffering ability by titrating with 0.005 M HCl and measuring pH by digital pH/Ion meter, used as a gold standart. The correlation were analyzed using the Spearman Rank Correlation Test, Cohen's Kappa coefficient and Pearson's Correlation test, p buffer capacity was found in 23.4% of cases, medium in 62.5%, and low in 14.1%. The Spearman Rank Correlation coefficient between the titration method and CRT Buffer test was 0.685 and the GC Saliva Check Buffer was 0.837. The Kappa coefficient for the CRT Buffer test was 0.508, while the coefficient for the GC Saliva Check Buffer was 0.752. The Pearson Correlation for the GC Saliva Check was 0.675. The difference is found in the buffer capacity at initial pH and at pH value 3. CONCLUSIONS. Both colorimetric tests correlate with the acid titration method in laboratory and are usable for saliva buffer capacity detection in dental offices. Buffer capacity detected in laboratory at different pH values can provide more information regarding caries risk.

  5. Systematic comparison of the human saliva and plasma proteomes

    Science.gov (United States)

    Yan, Weihong; Apweiler, Rolf; Balgley, Brian M.; Boontheung, Pinmanee; Bundy, Jonathan L.; Cargile, Benjamin J.; Cole, Steve; Fang, Xueping; Gonzalez-Begne, Mireya; Griffin, Timothy J.; Hagen, Fred; Hu, Shen; Wolinsky, Lawrence E.; Lee, Cheng S.; Malamud, Daniel; Melvin, James E.; Menon, Rajasree; Mueller, Michael; Qiao, Renli; Rhodus, Nelson L.; Sevinsky, Joel R.; States, David; Stephenson, James L.; Than, Shawn; Yates, John R.; Yu, Weixia; Xie, Hongwei; Xie, Yongming; Omenn, Gilbert S.; Loo, Joseph A.; Wong, David T.

    2009-01-01

    The proteome of human salivary fluid has the potential to open new doors for disease biomarker discovery. A recent study to comprehensively identify and catalog the human ductal salivary proteome led to the compilation of 1166 proteins. The protein complexity of both saliva and plasma is large, suggesting that a comparison of these two proteomes will provide valuable insight into their physiological significance and an understanding of the unique and overlapping disease diagnostic potential that each fluid provides. To create a more comprehensive catalog of human salivary proteins, we have first compiled an extensive list of proteins from whole saliva (WS) identified through MS experiments. The WS list is thereafter combined with the proteins identified from the ductal parotid, and submandibular and sublingual (parotid/SMSL) salivas. In parallel, a core dataset of the human plasma proteome with 3020 protein identifications was recently released. A total of 1939 nonredundant salivary proteins were compiled from a total of 19 474 unique peptide sequences identified from whole and ductal salivas; 740 out of the total 1939 salivary proteins were identified in both whole and ductal saliva. A total of 597 of the salivary proteins have been observed in plasma. Gene ontology (GO) analysis showed similarities in the distributions of the saliva and plasma proteomes with regard to cellular localization, biological processes, and molecular function, but revealed differences which may be related to the different physiological functions of saliva and plasma. The comprehensive catalog of the salivary proteome and its comparison to the plasma proteome provides insights useful for future study, such as exploration of potential biomarkers for disease diagnostics. PMID:19898684

  6. Expression of biological activity of draculin, the anticoagulant factor from vampire bat saliva, is strictly dependent on the appropriate glycosylation of the native molecule.

    Science.gov (United States)

    Fernandez, A Z; Tablante, A; Bartoli, F; Beguin, S; Hemker, H C; Apitz-Castro, R

    1998-10-23

    Draculin, a glycoprotein isolated from vampire bat (Desmodus rotundus) saliva, is a natural anticoagulant which inhibits activated coagulation factors IX (IXa) and X (Xa). The observation that under captivity conditions, the anticoagulant activity present in vampire bat saliva is dependent upon the salivation protocol, led us to investigate the possible relationship between the expression of biological activity of native draculin and the post-translational glycosylation of the protein backbone. Daily salivation of vampire bats yields a saliva that progressively decreases in anticoagulant activity, without any significant change in overall protein content, or in the amount of protein specifically recognized by a polyclonal anti-draculin antibody. Anticoagulant activity of the saliva is restored after a 4-day period of rest. Besides the marked difference in anticoagulant activity, purified native draculin, obtained from high- and low-activity saliva, shows significant differences in: (a) composition of the carbohydrate moiety, and (b) Glycosylation pattern. Furthermore, controlled chemical deglycosylation of native draculin, under conditions that do not affect the polypeptide backbone, progressively leads to complete loss of the biological activity. Our present results implicate that correct glycosylation of draculin is a seminal event for the expression of the biological activity of this glycoprotein.

  7. [Could saliva be used to detect HIV seroconversion?].

    Science.gov (United States)

    Sangaré, K A; Koffi, A R; Coulibaly, I M; Doulourou, C

    1997-01-01

    Blood is generally used for detection of antibodies associated with infections. However, removal of blood samples can be problematic and is painful for the patient. It requires suitable equipment and skilled staff, both of which may be expensive. Some patients refuse to allow removal of blood samples because they find it painful and traumatic. Removal of blood samples from children, newborns, immunocompromised or overweight subjects is often particularly difficult. In addition, some religions forbid the taking of blood samples. Thus, it is necessary to develop alternative, simple, painless methods of sampling body fluids that give results as accurate as those obtained with blood samples. Saliva has been suggested as a possible alternative. Epidemiological studies and other reports have shown that saliva may be of value for the detection of HIV antibodies. To evaluate the potential of saliva for detection of antibodies and seroconversion. Section I: Saliva and serum from 1,023 subjects, including 150 AIDS patients, 251 TB patients of known HIV status and 622 subjects from at-risk groups were tested. Sera from subjects with unknown HIV status were systematically tested by Abbott recombinant HIV1/HIV2 EIA. Section II: Saliva and sera were obtained from a population at risk of HIV infection. Two hundred and fifty consenting adults were found to be HIV-negative. Saliva was collected from each subject once per week and tested the same day with the Abbott test pack and Wellcozyme GACELISA. A spot of blood was also collected, dried and tested using the Wellcozyme GACELISA protocol. Whenever a positive result was obtained for any test, a blood sample was taken the same day or as soon as possible, for Western, blot analysis. Saliva was collected with an OMNISAL device (SDS, Vancouver, USA) placed under the tongue. The indicator turned blue when enough fluid had been collected. The device was then removed and the saliva placed in a tube with stabilized product. It was then

  8. [Detection of periodontal pathogens from saliva of type 2 diabetic patients in urban area of Beijing].

    Science.gov (United States)

    Liao, Yan-ting; He, Lu; Meng, Huan-xin; Li, Peng; Sha, Yue-qin; Wang, Xing-yu

    2013-03-01

    To investigate the prevalence of periodontal pathogens from saliva of patients with type 2 diabetes mellitus (T2DM), and to characterize the association between the glucose status and periodontal pathogens in oral cavity. All the subjects were hypertension patients under regular care at Beijing hypertension prevention and management institute. Whole unstimulated saliva samples were collected from 45 non-diabetic subjects (non-DM group), 80 well-controlled diabetic patients (DM-well group) and 100 poor-controlled diabetic patients (DM-poor group). DNA was extracted from the salivary deposition, Porphyromonas gingivalis (Pg), Tannerella forsythia (Tf) and Treponema denticola (Td) were detected by polymerase chain reaction (PCR) method based on 16SrRNA. Prevalence and quantity of the pathogens under different glucose states were compared and logistic regression model was set to analyze the factors related to each bacterium. The prevalence of Tf in DM-well group and DM-poor group was significantly lower than that of non-DM group [81% (65/80), 80% (80/100) vs 91% (41/45), P = 0.048], meanwhile the quantity of Tf was also lower than that of non-DM group [1.9(2.6), 2.1(5.3) vs 3.4(6.4)] (P > 0.05). With the worsening of glucose control, the quantity of Tf was declining (P = 0.032). However, the prevalence and the quantity of Pg, Td in 3 groups had no statistical differences (P > 0.05). After adjusting age, gender, number of missing teeth and other periodontal parameters, OR of having Tf in saliva from DM-well group and DM-poor group was 0.58 and 0.53, respectively. Abnormal blood glucose state may affect the colonization of Tf in oral cavity.

  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. Adverse affects of drugs on saliva and salivary glands

    Directory of Open Access Journals (Sweden)

    Vidhi Vinayak

    2013-01-01

    Full Text Available Saliva is the most valuable oral fluid is critical to the preservation and management of oral health. Saliva containing various organic and inorganic substances provides primary natural protection for teeth and soft tissues in the oral cavity assists in mastication, deglutition and digestion of food. The secretion of saliva can be affected due to various local and systemic causes. However if a patient is taking medication and has altered salivary secretion the differential diagnosis should include the possibility of an adverse drug reaction. The drugs may lead to alteration in the flow rate of saliva, which can be either increased or reduced, however certain drugs have been reported to cause change in the color of the saliva. Several drugs may lead to sialadenitis associated with altered salivary secretion. These symptoms may simulate systemic diseases, Hence oral physicians need to be vigilant in recognizing these adverse drug reactions in the patients and it is incumbent upon the practitioner to try to stay abreast of this ever evolving field especially as it relates to dental therapeutics.

  11. Method development for proteome stabilization in human saliva.

    Science.gov (United States)

    Xiao, Hua; Wong, David T W

    2012-04-13

    Human saliva is a biological fluid with emerging early detection and diagnostic potentials. However, the salivary proteome suffers from rapid degradation and thus compromises its translational and clinical utilities. Therefore, easy, reliable and practical methods are urgently required for the storage of human saliva samples. In this study, saliva samples from healthy subjects were collected and stored at room temperature (RT) and 4 °C for different lengths of time with and without specific protein stabilization treatments. SDS-PAGE was run to compare the protein profiling between samples. Reference proteins, β-actin and interleukin-1 β (IL1β), were chosen to evaluate salivary protein stability. Immunoassay was used for the detection of these target proteins. All data was compared with the positive control that had been kept at -80 °C. The results show that the salivary proteome that has been stored at 4 °C with added protease inhibitors was stable for approximately two weeks without significant degradation. By adding ethanol to the samples, the salivary proteome was stabilized at RT. After optimization, a simple, robust and convenient method is developed for the stabilization of proteins in human saliva that does not affect the downstream translational and clinical applications. The salivary proteome could be stabilized without significant degradation by adding ethanol at RT for about two weeks. This optimized method could greatly accelerate the clinical usage of saliva for future diagnosis. Copyright © 2012 Elsevier B.V. All rights reserved.

  12. Saliva: An emerging biofluid for early detection of diseases

    Science.gov (United States)

    Lee, Yu-Hsiang; Wong, David T.

    2010-01-01

    The capability to assess physiological states, detect morbidity initiation and progression, and monitor post-treatment therapeutic outcomes through a noninvasive approach is one of the most desirable goals for healthcare research and delivery. Saliva, a multi-constituent oral fluid, has high potential for the surveillance of general health and disease. To reach the above goal through saliva-based diagnostics, two prerequisites must be fulfilled: (1) discovering biomarker(s) for different diseases among the complicated components of saliva, and (2) advancing sensitivity and specificity of biomarker(s) through persistent development of technologies. Under the support and research blueprint initiated by the National Institute of Dental and Craniofacial Research (NIDCR), salivary diagnostics has not only steadily progressed with respect to accuracy and availability, but has also bridged up-to-date nanotechnology to expand the areas of application. With collective efforts over several years, saliva has been demonstrated to be a promising bodily fluid for early detection of diseases, and salivary diagnostics has exhibited tremendous potential in clinical applications. This review presents an overview of the value of saliva as a credible diagnostic tool, the discovery of salivary biomarkers, and the development of salivary diagnostics now and in the future. PMID:19824562

  13. 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.

  14. Deuterium equilibrium time in saliva of newborn infants.

    Science.gov (United States)

    Traver, Luis Angelo Marti; Martinez, Francisco Eulógio; Ferriolli, Eduardo; Marchini, Júlio Sérgio; Monteiro, Jacqueline Pontes; Pfrimer, Karina; Sanchez, Ana Paula Michelin; de Oliveira, Thais; Ducatti, Carlos; Camelo, José Simon

    2009-04-01

    There is an increasing interest about the use of stable isotopes for body composition analysis in pediatrics. To ensure the success of total body water analysis by the deuterium dilution method, it is fundamental to determine the equilibrium time (plateau) of deuterium in the body fluid studied. We report here the equilibration time of deuterium oxide in the saliva of newborns after oral intake of the isotope. Twenty healthy term newborn infants, 10 males and 10 females, were analyzed. Saliva was collected from each newborn before the oral administration of a 100 mg/kg dose of deuterium oxide (baseline sample) and then at 1-hour intervals for 5 hours after administration. Deuterium enrichment of saliva was determined by isotope ratio mass spectrometry according to the recommendations of the International Atomic Energy Agency. The plateau time of deuterium in saliva occurred 3 hours after oral administration of the stable isotope. These data are essential for further studies on the body composition of newborn infants. To the best of our knowledge, this is the first study regarding the equilibration time of deuterium in the saliva of term newborns.

  15. Saliva of obese patients – is it different?

    Directory of Open Access Journals (Sweden)

    Katarzyna Choromańska

    2015-01-01

    Full Text Available Obesity is a major public health concern that increases the risk of cardiovascular disease, type 2 diabetes and cancer. The incidence of obesity has increased significantly in recent years, not only in adults, but also in adolescents and children. This is evidenced by rapidly developing bariatric surgery, the most effective method of treating morbid obesity. Obesity is a multifactorial disease, and its pathogenesis is not completely understood. Numerous studies have been performed to clarify pathogenetic mechanisms, based mostly on blood and sometimes urine samples. Saliva is easily accessible and can be obtained non-invasively. Our aim was to review studies performed on saliva obtained from obese subjects in order to answer the title question.Obese people have different composition of salivary bacteria. Changes in the concentration of sialic acid, phosphorus and peroxidase activity as well as a lower flow rate of stimulated whole saliva promote dental caries and periodontal disease. Concentrations of salivary uric acid, endocannabinoids and CRP are increased in obesity and may provide a useful index of cardiometabolic risk. Assessment of fasting salivary ghrelin might facilitate choosing the best type of bariatric surgery for a specific patient. A significant decrease in salivary cortisol in women with morbid obesity also seems interesting.There is sufficient evidence to state that the saliva of obese and lean subjects is different. Saliva as an easily accessible research material seems promising, as shown by the few studies performed so far.

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

    Directory of Open Access Journals (Sweden)

    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.

  17. Potential applications of human saliva as diagnostic fluid.

    Science.gov (United States)

    Castagnola, M; Picciotti, P M; Messana, I; Fanali, C; Fiorita, A; Cabras, T; Calò, L; Pisano, E; Passali, G C; Iavarone, F; Paludetti, G; Scarano, E

    2011-12-01

    The use of human saliva as a diagnostic and prognostic fluid has until recently been somewhat disregarded. Although sample collection is non-invasive, physiological and genetic variations were largely responsible for its infrequent application in the past. Recently, several proteomic studies contributed to partial elucidation of the salivary proteome (more than 2400 protein components have been characterized), both in terms of composition, contributions to whole saliva and genetic/physiological variability. On this basis, is not too optimistic to believe that in the near future human saliva could become a relevant diagnostic fluid. In this review, the characterization by proteomic approaches of new salivary markers in oncology, head and neck carcinoma (oral cavity, oropharynx, larynx, and salivary glands), breast and gastric cancers, salivary gland function and disease, Sjögren syndrome, systemic sclerosis, dental and gingival pathology, systemic, psychiatric and neurological diseases, is described.

  18. Facilitated saliva secretion and reduced oral inflammation by a novel artificial saliva system in the treatment of salivary hypofunction.

    Science.gov (United States)

    Kang, Minkyung; Park, Hyounggeun; Jun, Joon-Ho; Son, Miwon; Kang, Myung Joo

    2017-01-01

    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.

  19. Helicobacter pylori detection in gastric biopsies, saliva and dental plaque of Brazilian dyspeptic patients

    Directory of Open Access Journals (Sweden)

    Lucas Trevizani Rasmussen

    2010-05-01

    Full Text Available Helicobacter pylori is an important human pathogen that causes chronic gastritis and is associated with the development of peptic ulcer disease and gastric malignancies. The oral cavity has been implicated as a potential H. pylori reservoir and may therefore be involved in the reinfection of the stomach, which can sometimes occur following treatment of an H. pylori infection. The objectives of this paper were (i to determine the presence of H. pylori in the oral cavity and (ii to examine the relationship between oral H. pylori and subsequent gastritis. Gastric biopsies, saliva samples and dental plaques were obtained from 78 dyspeptic adults. DNA was extracted and evaluated for the presence of H. pylori using polymerase chain reaction and Southern blotting methods. Persons with gastritis were frequently positive for H. pylori in their stomachs (p < 0.0001 and there was a statistically significant correlation between the presence of H. pylori in gastric biopsies and the oral cavity (p < 0.0001. Our results suggest a relationship between gastric infection and the presence of this bacterium in the oral cavity. Despite this, H. pylori were present in the oral cavity with variable distribution between saliva and dental plaques, suggesting the existence of a reservoir for the species and a potential association with gastric reinfection.

  20. Rapid Antemortem Detection of CWD Prions in Deer Saliva

    Science.gov (United States)

    Haley, Nicholas J.; Denkers, Nathaniel D.; Nalls, Amy V.; Mathiason, Candace K.; Caughey, Byron; Hoover, Edward A.

    2013-01-01

    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. PMID:24040235

  1. Quantitative proteomic analysis of the fall armyworm saliva.

    Science.gov (United States)

    Acevedo, Flor E; Stanley, Bruce A; Stanley, Anne; Peiffer, Michelle; Luthe, Dawn S; Felton, Gary W

    2017-07-01

    Lepidopteran larvae secrete saliva on plant tissues during feeding. Components in the saliva may aid in food digestion, whereas other components are recognized by plants as cues to elicit defense responses. Despite the ecological and economical importance of these plant-feeding insects, knowledge of their saliva composition is limited to a few species. In this study, we identified the salivary proteins of larvae of the fall armyworm (FAW), Spodoptera frugiperda; determined qualitative and quantitative differences in the salivary proteome of the two host races-corn and rice strains-of this insect; and identified changes in total protein concentration and relative protein abundance in the saliva of FAW larvae associated with different host plants. Quantitative proteomic analyses were performed using labeling with isobaric tags for relative and absolute quantification followed by liquid chromatography-tandem mass spectrometry. In total, 98 proteins were identified (>99% confidence) in the FAW saliva. These proteins were further categorized into five functional groups: proteins potentially involved in (1) plant defense regulation, (2) herbivore offense, (3) insect immunity, (4) detoxification, (5) digestion, and (6) other functions. Moreover, there were differences in the salivary proteome between the FAW strains that were identified by label-free proteomic analyses. Thirteen differentially identified proteins were present in each strain. There were also differences in the relative abundance of eleven salivary proteins between the two FAW host strains as well as differences within each strain associated with different diets. The total salivary protein concentration was also different for the two strains reared on different host plants. Based on these results, we conclude that the FAW saliva contains a complex mixture of proteins involved in different functions that are specific for each strain and its composition can change plastically in response to diet type

  2. 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.

  3. 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

  4. 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 (R2=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 treatment

  5. 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

  6. Prevalence of Pseudomonas aeruginosa and Acinetobacter spp. in subgingival biofilm and saliva of subjects with chronic periodontal infection

    Science.gov (United States)

    Souto, Renata; Silva-Boghossian, Carina M.; Colombo, Ana Paula Vieira

    2014-01-01

    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 and > 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 oral microbiota of CP. Poor oral hygiene, smoking and the presence of P. aeruginosa are strongly associated with periodontitis. PMID:25242933

  7. Prevalence of Pseudomonas aeruginosa and Acinetobacter spp. in subgingival biofilm and saliva of subjects with chronic periodontal infection.

    Science.gov (United States)

    Souto, Renata; Silva-Boghossian, Carina M; Colombo, Ana Paula Vieira

    2014-01-01

    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 and > 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 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.

  8. Sample Stability and Protein Composition of Saliva: Implications for Its Use as a Diagnostic Fluid

    OpenAIRE

    Han Roelofsen; Vonk, Roel J.; Desiree Weening; Gloria Alvarez-Llamas; de Vries, Marcel P.; Diederik Esser

    2008-01-01

    Saliva is an easy accessible plasma ultra-filtrate. Therefore, saliva can be an attractive alternative to blood for measurement of diagnostic protein markers. Our aim was to determine stability and protein composition of saliva. Protein stability at room temperature was examined by incubating fresh whole saliva with and without inhibitors of proteases and bacterial metabolism followed by Surface Enhanced Laser Desorption/Ionization (SELDI) analyses. Protein composition was determined by sodiu...

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

    OpenAIRE

    Maya Rashkova

    2012-01-01

    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 con...

  10. Artificial saliva effect on toxic substances release from acrylic resins.

    Science.gov (United States)

    Kostić, Milena; Krunić, Nebojša; Najman, Stevo; Nikolić, Ljubiša; Nikolić, Vesna; Rajković, Jelena; Petrović, Milica; Igić, Marko; Ignjatović, Aleksandra

    2015-10-01

    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. Storage of acrylic samples in two models of artificial saliva were performed in a water bath at the temperature of 37 +/- 1 degrees C. Analysis of the surface structure of samples was carned out using scanning electronic microscopy analysis immedidtely 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. 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. In order to improve biological features of acrylic resin materials, it was recommended that dentures lined with soft or hard cold-polymerized 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.

  11. 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

  12. Saliva pH affects the sweetness sense.

    Science.gov (United States)

    Aoyama, Ken-Ichi; Okino, Yuichiro; Yamazaki, Hiroshi; Kojima, Rena; Uchibori, Masahiro; Nakanishi, Yaushiro; Ota, Yoshihide

    2017-03-01

    The aim of this study was to establish a prediction system for taste sense according to the biochemical data of saliva. The present study included 100 participants ages ≥20 y without physical, mental, or dental disabilities. Saliva samples were collected from the participants and subjected to biochemical analyses. Taste examination (sweetness, saltiness, sourness, and bitterness) was performed using the dropped disk method. Correlation analysis and multiple regression analysis were performed between the taste sense properties and biochemical data of saliva. Multiple regression analysis demonstrated that sweetness sensitivity (in which a higher score indicates lower sensitivity) was significantly affected by various biochemical properties, with the strongest influence being pH. The following prediction equation was determined: Sweetness sensitivity = 1.38 + (-0.12 × low pH [1: If pH 7.3, 0: otherwise]) + (0.04 × Fe [μg/dL]). Analysis of variance showed an overall significant effect of these variables on sweetness sensitivity (R2 = 0.74; P < 0.01). Saliva pH most strongly affects the sweetness sensitivity. This prediction can be used for evaluations of variations in dietary choices and to help individuals make healthy food choices to maintain health. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. The use of saliva markers in psychobiology: mechanisms and methods

    NARCIS (Netherlands)

    Bosch, J.A.; Ligtenberg, A.J.M.; Veerman, E.C.I.

    2014-01-01

    In the social sciences, the use of saliva parameters has greatly expanded in recent years from the measurement of steroid hormones, like cortisol, and now includes a wide range of biochemical parameters. These salivary constituents can be broadly classified into two groups: (1) constituents that

  14. Does menstrual cycle effect buffer capacity of stimulated saliva?

    Science.gov (United States)

    Dural, Sema; Cağirankaya, Leyla Berna

    2007-09-01

    The present study was performed to analyze buffer capacity (BC) and flow rate of stimulated saliva during menstrual cycle. Two salivary samples were taken from 17 subjects during the menstrual cycle. BC was determined according to electrometric method. Both variables showed no hormone dependency. The results suggest that the salivary protection against acid attacks is constant in healthy nonpregnant women.

  15. DETERMINATION OF CHLORHEXIDINE IN SALIVA AND IN AQUEOUS-SOLUTIONS

    NARCIS (Netherlands)

    de Vries, J.; Ruben, J; Arends, J.

    1991-01-01

    A new method is presented for the determination of chlorhexidine in centrifuged saliva and in aqueous solutions by means of fluorescence spectroscopy. The method relies on complex formation between chlorhexidine and eosin. The fluorescence value of the chlorhexidine-eosin system decreases with

  16. Cell-derived vesicles exposing coagulant tissue factor in saliva

    NARCIS (Netherlands)

    Berckmans, René J.; Sturk, Auguste; van Tienen, Laurens M.; Schaap, Marianne C. L.; Nieuwland, Rienk

    2011-01-01

    On vascular damage, coagulation is initiated by extravascular tissue factor (TF). Intravascular TF, which is present on circulating cell-derived vesicles, is non-coagulant under physiologic conditions but prothrombotic under pathologic conditions. Human saliva triggers coagulation, but the mechanism

  17. Saliva-catalyzed hydrolysis of a ketobemidone ester prodrug

    DEFF Research Database (Denmark)

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

    1992-01-01

    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....

  18. Fourier Transform Infrared Spectroscopy and Photoacoustic Spectroscopy for Saliva Analysis.

    Science.gov (United States)

    Mikkonen, Jopi J W; Raittila, Jussi; Rieppo, Lassi; Lappalainen, Reijo; Kullaa, Arja M; Myllymaa, Sami

    2016-09-01

    Saliva provides a valuable tool for assessing oral and systemic diseases, but concentrations of salivary components are very small, calling the need for precise analysis methods. In this work, Fourier transform infrared (FT-IR) spectroscopy using transmission and photoacoustic (PA) modes were compared for quantitative analysis of saliva. The performance of these techniques was compared with a calibration series. The linearity of spectrum output was verified by using albumin-thiocyanate (SCN(-)) solution at different SCN(-) concentrations. Saliva samples used as a comparison were obtained from healthy subjects. Saliva droplets of 15 µL were applied on the silicon sample substrate, 6 drops for each specimen, and dried at 37 ℃ overnight. The measurements were carried out using an FT-IR spectrometer in conjunction with an accessory unit for PA measurements. The findings with both transmission and PA modes mirror each other. The major bands presented were 1500-1750 cm(-1) for proteins and 1050-1200 cm(-1) for carbohydrates. In addition, the distinct spectral band at 2050 cm(-1) derives from SCN(-) anions, which is converted by salivary peroxidases to hypothiocyanate (OSCN(-)). The correlation between the spectroscopic data with SCN(-) concentration (r > 0.990 for transmission and r = 0.967 for PA mode) was found to be significant (P < 0.01), thus promising to be utilized in future applications. © The Author(s) 2016.

  19. Variability of flow rate when collecting stimulated human parotid saliva

    NARCIS (Netherlands)

    Burlage, FR; Pijpe, J; Coppes, RP; Hemels, MEW; Meertens, H; Canrinus, A; Vissink, A

    2005-01-01

    The aim of this study was to estimate the accuracy and reproducibility of citric-acid-stimulated parotid saliva sampling. In healthy volunteers a strong correlation (r(2) = 0.79) between flow rates from the left and right parotid gland was observed. In patients with Sjogren's syndrome this

  20. Saliva secretion rate and acidity in a group of physically disabled older care home residents

    NARCIS (Netherlands)

    Putten, G.J. van der; Brand, H.S.; De Visschere, L.M.; Schols, J.M.; Baat, C. de

    2013-01-01

    A growing number of older people have teeth, which are vulnerable to oral diseases. To maintain good oral health, an adequate amount of saliva should be secreted and the saliva should possess adequate buffer capacity. The study aim was to investigate the associations of saliva secretion rate and

  1. Sample Stability and Protein Composition of Saliva : Implications for Its Use as a Diagnostic Fluid

    NARCIS (Netherlands)

    Esser, Diederik; Alvarez-Llamas, Gloria; de Vries, Marcel P; Weening, Desiree; Vonk, Roel J; Roelofsen, Han

    2008-01-01

    Saliva is an easy accessible plasma ultra-filtrate. Therefore, saliva can be an attractive alternative to blood for measurement of diagnostic protein markers. Our aim was to determine stability and protein composition of saliva. Protein stability at room temperature was examined by incubating fresh

  2. A lacticin 3147 enriched food ingredient reduces Streptococcus mutans isolated from the human oral cavity in saliva.

    Science.gov (United States)

    O'Connor, E B; O'Riordan, B; Morgan, S M; Whelton, H; O'Mullane, D M; Ross, R P; Hill, C

    2006-06-01

    To isolate and characterise Streptococcus mutans from Irish saliva samples and to assess their sensitivity to a food-grade preparation of the lantibiotic, lacticin 3147, produced by Lactococcus lactis DPC3147. Saliva samples collected from children with varying oral health status were screened on Mitis Salivarius agar for the presence of pathogenic streptococci. Following selective plating, 16S rDNA sequencing and Pulsed Field Gel Electrophoresis (PFGE), 15 distinct strains of Strep. mutans were identified. These were grouped according to their relative sensitivity to lacticin 3147 which ranged from 0.78 to 6.25%; relative to a sensitive indicator strain, Lactococcus lactis ssp. lactis HP. Inhibition of indicator Strep. mutans strains from sensitive, intermediate and tolerant groupings were assessed in microtitre plate assays with increasing concentrations of lacticin 3147. The concentration of lacticin 3147 required to give 50% growth inhibition correlated with their relative sensitivities (as assayed by well diffusion methodology) and ranged from 1280 to 5120 AU ml(-1). Concentrated preparations of lacticin 3147 caused a rapid killing of Strep. mutans strains in broth. Moreover, in human saliva deliberately spiked with Strep. mutans, the pathogen was eliminated (initial inoculum of 10(5)) in the presence of 40,000 AU ml(-1) of lacticin 3147. Furthermore, a food-grade lacticin 3147 spray dried powder ingredient was assessed for the inhibition of Strep. mutans in human saliva, spiked with a strain of intermediate sensitivity, resulting in up to a 4-log reduction in counts after 20 min. A food grade preparation of lacticin 3147 was effective in the inhibition of oral Strep. mutans. The inhibition of oral streptococci by food grade preparations of lacticin 3147 may offer novel opportunities for the development of lacticin 3147 as an anti-cariogenic agent particularly in the area of functional foods for the improvement of oral health.

  3. RNA interference inhibits herpes simplex virus type 1 isolated from saliva samples and mucocutaneous lesions.

    Science.gov (United States)

    Silva, Amanda Perse da; Lopes, Juliana Freitas; Paula, Vanessa Salete de

    2014-01-01

    The aim of this study was to evaluate the use of RNA interference to inhibit herpes simplex virus type-1 replication in vitro. For herpes simplex virus type-1 gene silencing, three different small interfering RNAs (siRNAs) targeting the herpes simplex virus type-1 UL39 gene (sequence si-UL 39-1, si-UL 39-2, and si-UL 39-3) were used, which encode the large subunit of ribonucleotide reductase, an essential enzyme for DNA synthesis. Herpes simplex virus type-1 was isolated from saliva samples and mucocutaneous lesions from infected patients. All mucocutaneous lesions' samples were positive for herpes simplex virus type-1 by real-time PCR and by virus isolation; all herpes simplex virus type-1 from saliva samples were positive by real-time PCR and 50% were positive by virus isolation. The levels of herpes simplex virus type-1 DNA remaining after siRNA treatment were assessed by real-time PCR, whose results demonstrated that the effect of siRNAs on gene expression depends on siRNA concentration. The three siRNA sequences used were able to inhibit viral replication, assessed by real-time PCR and plaque assays and among them, the sequence si-UL 39-1 was the most effective. This sequence inhibited 99% of herpes simplex virus type-1 replication. The results demonstrate that silencing herpes simplex virus type-1 UL39 expression by siRNAs effectively inhibits herpes simplex virus type-1 replication, suggesting that siRNA based antiviral strategy may be a potential therapeutic alternative. Copyright © 2014. Published by Elsevier Editora Ltda.

  4. In silico target network analysis of de novo-discovered, tick saliva-specific microRNAs reveals important combinatorial effects in their interference with vertebrate host physiology.

    Science.gov (United States)

    Hackenberg, Michael; Langenberger, David; Schwarz, Alexandra; Erhart, Jan; Kotsyfakis, Michail

    2017-08-01

    The hard tick Ixodes ricinus is an important disease vector whose salivary secretions mediate blood-feeding success on vertebrate hosts, including humans. Here we describe the expression profiles and downstream analysis of de novo-discovered microRNAs (miRNAs) expressed in I. ricinus salivary glands and saliva. Eleven tick-derived libraries were sequenced to produce 67,375,557 Illumina reads. De novo prediction yielded 67 bona fide miRNAs out of which 35 are currently not present in miRBase. We report for the first time the presence of microRNAs in tick saliva, obtaining furthermore molecular indicators that those might be of exosomal origin. Ten out of these microRNAs are at least 100 times more represented in saliva. For the four most expressed microRNAs from this subset, we analyzed their combinatorial effects upon their host transcriptome using a novel in silico target network approach. We show that only the inclusion of combinatorial effects reveals the functions in important pathways related to inflammation and pain sensing. A control set of highly abundant microRNAs in both saliva and salivary glands indicates no significant pathways and a far lower number of shared target genes. Therefore, the analysis of miRNAs from pure tick saliva strongly supports the hypothesis that tick saliva miRNAs can modulate vertebrate host homeostasis and represents the first direct evidence of tick miRNA-mediated regulation of vertebrate host gene expression at the tick-host interface. As such, the herein described miRNAs may support future drug discovery and development projects that will also experimentally question their predicted molecular targets in the vertebrate host. © 2017 Hackenberg et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

  5. Saliva and Serum Protein Exchange at the Tooth Enamel Surface.

    Science.gov (United States)

    Heller, D; Helmerhorst, E J; Oppenheim, F G

    2017-04-01

    The acquired enamel pellicle is an oral, fluid-derived protein layer that forms on the tooth surface. It is a biologically and clinically important integument that protects teeth against enamel demineralization, and abrasion. Tooth surfaces are exposed to different proteinaceous microenvironments depending on the enamel location. For instance, tooth surfaces close to the gingival sulcus contact serum proteins that emanate via this sulcus, which may impact pellicle composition locally. The aims of this study were to define the major salivary and serum components that adsorb to hydroxyapatite, to study competition among them, and to obtain preliminary evidence in an in vivo saliva/serum pellicle model. Hydroxyapatite powder was incubated with saliva and serum, and the proteins that adsorbed were identified by mass spectrometry. To study competition, saliva and serum proteins were labeled with CyDyes, mixed in various proportions, and incubated with hydroxyapatite. In vivo competition was assessed using a split-mouth design, with half the buccal tooth surfaces coated with serum and the other half with saliva. After exposure to the oral environment for 0 min, 30 min and 2 h, the pellicles were analyzed by SDS-PAGE. In pure saliva- or serum-derived pellicles, 82 and 84 proteins were identified, respectively. When present concomitantly, salivary protein adsorbers effectively competed with serum protein adsorbers for the hydroxyapatite surface. Specifically, acidic proline-rich protein, cystatin, statherin and protein S100-A9 proteins competed off apolipoproteins, complement C4-A, haptoglobin, transthyretin and serotransferrin. In vivo evidence further supported the replacement of serum proteins by salivary proteins. In conclusion, although significant numbers of serum proteins emanate from the gingival sulcus, their ability to participate in dental pellicle formation is likely reduced in the presence of strong salivary protein adsorbers. The functional properties of the

  6. The impact of DNA input amount and DNA source on the performance of whole-exome sequencing in cancer epidemiology.

    Science.gov (United States)

    Zhu, Qianqian; Hu, Qiang; Shepherd, Lori; Wang, Jianmin; Wei, Lei; Morrison, Carl D; Conroy, Jeffrey M; Glenn, Sean T; Davis, Warren; Kwan, Marilyn L; Ergas, Isaac J; Roh, Janise M; Kushi, Lawrence H; Ambrosone, Christine B; Liu, Song; Yao, Song

    2015-08-01

    Whole-exome sequencing (WES) has recently emerged as an appealing approach to systematically study coding variants. However, the requirement for a large amount of high-quality DNA poses a barrier that may limit its application in large cancer epidemiologic studies. We evaluated the performance of WES with low input amount and saliva DNA as an alternative source material. Five breast cancer patients were randomly selected from the Pathways Study. From each patient, four samples, including 3 μg, 1 μg, and 0.2 μg blood DNA and 1 μg saliva DNA, were aliquoted for library preparation using the Agilent SureSelect Kit and sequencing using Illumina HiSeq2500. Quality metrics of sequencing and variant calling, as well as concordance of variant calls from the whole exome and 21 known breast cancer genes, were assessed by input amount and DNA source. There was little difference by input amount or DNA source on the quality of sequencing and variant calling. The concordance rate was about 98% for single-nucleotide variant calls and 83% to 86% for short insertion/deletion calls. For the 21 known breast cancer genes, WES based on low input amount and saliva DNA identified the same set variants in samples from a same patient. Low DNA input amount, as well as saliva DNA, can be used to generate WES data of satisfactory quality. Our findings support the expansion of WES applications in cancer epidemiologic studies where only low DNA amount or saliva samples are available. ©2015 American Association for Cancer Research.

  7. Recovery of DNA and fingermarks following deployment of render-safe tools for vehicle-borne improvised explosive devices (VBIED).

    Science.gov (United States)

    Ramasamy, S; Houspian, A; Knott, F

    2011-07-15

    Improvised explosive devices (IED) are responsible for a significant proportion of combat and civilian deaths around the world. Given the ease with which IEDs can be made, the large quantity of explosive which can be contained within or on a vehicle, and the use of VBIED in the past (for example the 2002 Bali bombing) in terrorist activities, VBIED are an ongoing concern for Defence and law enforcement agencies. Fingermark and DNA analyses are routinely used by police and forensic analysts to identify suspects involved in illegal activities. There is limited information available on the feasibility of obtaining fingermarks, fibres, hair and DNA samples following an explosive incident, or a situation whereby an IED has been rendered safe following the utilisation of an appropriate defeat or render-safe tool. The main objective of this study was to determine if fingermarks and/or DNA (from saliva and hair samples) placed on the interior and exterior of road vehicles, and on inanimate objects (such as plastic or glass bottles), are able to be obtained and analysed following the use of a vehicle-borne IED (VBIED) render-safe tool on a vehicle containing simulated explosives. The identification of fingermarks on the exterior (67.2±8.5%) and interior (43.8±17.8%) of the vehicles was possible following the use of the render-safe tool, though this was more challenging in the latter than the former. Fingermarks were also able to be identified from both plastic and glass bottles placed inside the vehicles. Polymerase chain reaction (PCR) techniques yielded DNA profiles that were able to be identified from saliva and hair samples. These preliminary results suggest that both fingermarks and DNA profiles, obtained from vehicles that have been subjected to a VBIED render-safe tool, may be used to identify persons of interest. Crown Copyright © 2011. Published by Elsevier Ireland Ltd. All rights reserved.

  8. Comparative analysis of bacterial profiles in unstimulated and stimulated saliva samples

    Directory of Open Access Journals (Sweden)

    Daniel Belstrøm

    2016-03-01

    Full Text Available Background and objective: The microbial profiles of stimulated saliva samples have been shown to differentiate between patients with periodontitis, patients with dental caries, and orally healthy individuals. Saliva was stimulated to allow for easy and rapid collection; however, microbial composition may not reflect the more natural, unstimulated state. The purpose of this study was to validate whether stimulated saliva is an adequate surrogate for unstimulated saliva in determining salivary microbiomes. Design: Unstimulated (n=20 and stimulated (n=20 saliva samples were collected from 20 orally and systemically healthy, non-smoking participants. Salivary bacterial profiles were analyzed by means of the Human Oral Microbe Identification using Next Generation Sequencing (HOMINGS, and statistical analysis was performed using Mann–Whitney test with Benjamini–Hochberg's correction for multiple comparison, cluster analysis, principal component analysis, and correspondence analysis. Results: From a total of 40 saliva samples, 496 probe targets were identified with a mean number of targets per sample of 203 (range: 146–303, and a mean number of probe targets of 206 and 200 in unstimulated and stimulated saliva samples, respectively (p=0.62. Based on all statistical methods used for this study, the microbial profiles of unstimulated and stimulated saliva samples collected from the same person were not statistically significantly different. Conclusions: Analysis of bacterial salivary profiles in unstimulated and stimulated saliva samples collected from the same individual showed comparable results. Thus, the results verify that stimulated saliva is an adequate surrogate of unstimulated saliva for microbiome-related studies.

  9. Insights into the saliva of the brown marmorated stink bug Halyomorpha halys (Hemiptera: Pentatomidae).

    Science.gov (United States)

    Peiffer, Michelle; Felton, Gary W

    2014-01-01

    We examined the salivary gland structure of the brown marmorated stink bug (Pentatomidae: Halyomorpha halys) and developed methods for independent collection of watery saliva and sheath saliva. This stink bug has become a serious invasive pest of agriculture in the United States and its saliva is largely responsible for the damage it causes. We determined by protein gel analysis and shotgun proteomics that the suite of proteins comprising the sheath and watery saliva are very distinct. Our results indicate that a substantial amount of sheath proteins are derived from tomato when stink bugs feed on tomato fruit. Consequently, the sheath saliva is comprised of both insect and plant-derived proteins. Both sheath and watery saliva possessed amylase activities, but polyphenol oxidase and glucose oxidase activities were not detected in either saliva. Peroxidase activity was only detected in salivary sheaths, but only when stink bugs fed on tomato. Proteomic analysis indicated that the peroxidase was likely of plant origin. We also determined that sheath saliva, but not watery saliva elicited the jasmonate inducible defense gene proteinase inhibitor 2 (Pin2), but this induction was only observed when sheaths had been collected from tomato. This indicates that the eliciting factor of the saliva is likely of plant origin. Lastly, neither watery or sheath saliva affected the expression of the salicylate inducible gene pathogenesis related gene (Pr1a-P4).

  10. Insights into the saliva of the brown marmorated stink bug Halyomorpha halys (Hemiptera: Pentatomidae.

    Directory of Open Access Journals (Sweden)

    Michelle Peiffer

    Full Text Available We examined the salivary gland structure of the brown marmorated stink bug (Pentatomidae: Halyomorpha halys and developed methods for independent collection of watery saliva and sheath saliva. This stink bug has become a serious invasive pest of agriculture in the United States and its saliva is largely responsible for the damage it causes. We determined by protein gel analysis and shotgun proteomics that the suite of proteins comprising the sheath and watery saliva are very distinct. Our results indicate that a substantial amount of sheath proteins are derived from tomato when stink bugs feed on tomato fruit. Consequently, the sheath saliva is comprised of both insect and plant-derived proteins. Both sheath and watery saliva possessed amylase activities, but polyphenol oxidase and glucose oxidase activities were not detected in either saliva. Peroxidase activity was only detected in salivary sheaths, but only when stink bugs fed on tomato. Proteomic analysis indicated that the peroxidase was likely of plant origin. We also determined that sheath saliva, but not watery saliva elicited the jasmonate inducible defense gene proteinase inhibitor 2 (Pin2, but this induction was only observed when sheaths had been collected from tomato. This indicates that the eliciting factor of the saliva is likely of plant origin. Lastly, neither watery or sheath saliva affected the expression of the salicylate inducible gene pathogenesis related gene (Pr1a-P4.

  11. Plants can benefit from herbivory: stimulatory effects of sheep saliva on growth of Leymus chinensis.

    Directory of Open Access Journals (Sweden)

    Jushan Liu

    Full Text Available BACKGROUND: Plants and herbivores can evolve beneficial interactions. Growth factors found in animal saliva are probably key factors underlying plant compensatory responses to herbivory. However, there is still a lack of knowledge about how animal saliva interacts with herbivory intensities and how saliva can mobilize photosynthate reserves in damaged plants. METHODOLOGY/PRINCIPAL FINDINGS: The study examined compensatory responses to herbivory and sheep saliva addition for the grass species Leymus chinensis in three experiments over three years. The first two experiments were conducted in a factorial design with clipping (four levels in 2006 and five in 2007 and two saliva treatment levels. The third experiment examined the mobilization and allocation of stored carbohydrates following clipping and saliva addition treatments. Animal saliva significantly increased tiller number, number of buds, and biomass, however, there was no effect on height. Furthermore, saliva effects were dependent on herbivory intensities, associated with meristem distribution within perennial grass. Animal saliva was found to accelerate hydrolyzation of fructans and accumulation of glucose and fructose. CONCLUSIONS/SIGNIFICANCE: The results demonstrated a link between saliva and the mobilization of carbohydrates following herbivory, which is an important advance in our understanding of the evolution of plant responses to herbivory. Herbivory intensity dependence of the effects of saliva stresses the significance of optimal grazing management.

  12. Plants can benefit from herbivory: stimulatory effects of sheep saliva on growth of Leymus chinensis.

    Science.gov (United States)

    Liu, Jushan; Wang, Ling; Wang, Deli; Bonser, Stephen P; Sun, Fang; Zhou, Yifa; Gao, Ying; Teng, Xing

    2012-01-01

    Plants and herbivores can evolve beneficial interactions. Growth factors found in animal saliva are probably key factors underlying plant compensatory responses to herbivory. However, there is still a lack of knowledge about how animal saliva interacts with herbivory intensities and how saliva can mobilize photosynthate reserves in damaged plants. The study examined compensatory responses to herbivory and sheep saliva addition for the grass species Leymus chinensis in three experiments over three years. The first two experiments were conducted in a factorial design with clipping (four levels in 2006 and five in 2007) and two saliva treatment levels. The third experiment examined the mobilization and allocation of stored carbohydrates following clipping and saliva addition treatments. Animal saliva significantly increased tiller number, number of buds, and biomass, however, there was no effect on height. Furthermore, saliva effects were dependent on herbivory intensities, associated with meristem distribution within perennial grass. Animal saliva was found to accelerate hydrolyzation of fructans and accumulation of glucose and fructose. The results demonstrated a link between saliva and the mobilization of carbohydrates following herbivory, which is an important advance in our understanding of the evolution of plant responses to herbivory. Herbivory intensity dependence of the effects of saliva stresses the significance of optimal grazing management.

  13. Plants Can Benefit from Herbivory: Stimulatory Effects of Sheep Saliva on Growth of Leymus chinensis

    Science.gov (United States)

    Liu, Jushan; Wang, Ling; Wang, Deli; Bonser, Stephen P.; Sun, Fang; Zhou, Yifa; Gao, Ying; Teng, Xing

    2012-01-01

    Background Plants and herbivores can evolve beneficial interactions. Growth factors found in animal saliva are probably key factors underlying plant compensatory responses to herbivory. However, there is still a lack of knowledge about how animal saliva interacts with herbivory intensities and how saliva can mobilize photosynthate reserves in damaged plants. Methodology/Principal Findings The study examined compensatory responses to herbivory and sheep saliva addition for the grass species Leymus chinensis in three experiments over three years. The first two experiments were conducted in a factorial design with clipping (four levels in 2006 and five in 2007) and two saliva treatment levels. The third experiment examined the mobilization and allocation of stored carbohydrates following clipping and saliva addition treatments. Animal saliva significantly increased tiller number, number of buds, and biomass, however, there was no effect on height. Furthermore, saliva effects were dependent on herbivory intensities, associated with meristem distribution within perennial grass. Animal saliva was found to accelerate hydrolyzation of fructans and accumulation of glucose and fructose. Conclusions/Significance The results demonstrated a link between saliva and the mobilization of carbohydrates following herbivory, which is an important advance in our understanding of the evolution of plant responses to herbivory. Herbivory intensity dependence of the effects of saliva stresses the significance of optimal grazing management. PMID:22235277

  14. Insights into the Saliva of the Brown Marmorated Stink Bug Halyomorpha halys (Hemiptera: Pentatomidae)

    Science.gov (United States)

    Peiffer, Michelle; Felton, Gary W.

    2014-01-01

    We examined the salivary gland structure of the brown marmorated stink bug (Pentatomidae: Halyomorpha halys) and developed methods for independent collection of watery saliva and sheath saliva. This stink bug has become a serious invasive pest of agriculture in the United States and its saliva is largely responsible for the damage it causes. We determined by protein gel analysis and shotgun proteomics that the suite of proteins comprising the sheath and watery saliva are very distinct. Our results indicate that a substantial amount of sheath proteins are derived from tomato when stink bugs feed on tomato fruit. Consequently, the sheath saliva is comprised of both insect and plant-derived proteins. Both sheath and watery saliva possessed amylase activities, but polyphenol oxidase and glucose oxidase activities were not detected in either saliva. Peroxidase activity was only detected in salivary sheaths, but only when stink bugs fed on tomato. Proteomic analysis indicated that the peroxidase was likely of plant origin. We also determined that sheath saliva, but not watery saliva elicited the jasmonate inducible defense gene proteinase inhibitor 2 (Pin2), but this induction was only observed when sheaths had been collected from tomato. This indicates that the eliciting factor of the saliva is likely of plant origin. Lastly, neither watery or sheath saliva affected the expression of the salicylate inducible gene pathogenesis related gene (Pr1a-P4). PMID:24586332

  15. Saliva: A tool in assessing glucose levels in Diabetes Mellitus.

    Science.gov (United States)

    Satish, B N V S; Srikala, P; Maharudrappa, B; Awanti, Sharanabasappa M; Kumar, Prashant; Hugar, Deepa

    2014-04-01

    Diabetes mellitus is a metabolic disorder affecting people worldwide, which require constant monitoring of their glucose levels. Commonly employed procedures include collection of blood or urine samples causing discomfort to the patients. Hence the need for an alternative non invasive technique is required to monitor glucose levels. Saliva present in the oral cavity not only maintains the health of the oral cavity but plays a important role in diagnosis of cancers of the oral cavity, periodontal diseases, HIV, heart diseases etc. The aim of the present study was undertaken to correlate the glucose levels in saliva and blood of diabetic and healthy non diabetic individuals and to determine the efficacy of saliva as a diagnostic tool. A total of 30 individuals of which 20 patients were diabetic patients and on medication and 10 patients were healthy non diabetic individuals were included in the study. Blood and saliva were collected under resting conditions and were subjected to glucose estimation. Salivary and blood glucose concentrations were determined in non diabetic healthy individuals (n=10) and Type II Diabetes mellitus patients (n=20). Glycosylated haemoglobin A1c was also determined in both Type II diabetic patients and Control group and a significant correlation (r=0.73) and (r=0.46) was found between HbA1c and serum glucose concentrations in diabetic and control group respectively. A significant correlation (r=0.54) and (r=0.45) was found between fasting blood glucose and fasting salivary glucose for diabetic group and control group respectively. A positive correlation (r=0.39) and (r=0.38) was found between fasting salivary glucose and HbA1c for diabetic and control group respectively. These findings suggest that the saliva can be used in the assessment of the blood glucose concentration in diabetes mellitus patients. How to cite the article: Satish BN, Srikala P, Maharudrappa B, Awanti M, Kumar P, Hugar D. Saliva: A tool in assessing glucose levels in

  16. The influence of tooth brushing time over saliva buffering capacity

    Directory of Open Access Journals (Sweden)

    Sri Mulyanti

    2014-11-01

    Full Text Available Saliva gives a considerable influence against the growth of dental caries as a natural defense against caries. the things very important about saliva are its flow rate and buffering capacity. the decrease in saliva flow rate might cause food retention that furthermore would turn into dental plaques, meanwhile it’s buffering capacity will play a considerable role in maintaining the saliva’s pH and remineralization process of the teeth. One of the mechanisms which are considered to be effective in preventing dental caries is teeth brushing which could change the pH of 5,6 to a normal level. And the right time of teeth brushing will provide an optimal result.The study aims to reveal the influence of teeth brushing time against saliva buffering capacity. The study is an analytic study using a quasi-experimental design. The samples of the study are 20 (twenty students of dentistry in Health Ministry of Bandung which was purposively selected the sample is divided into 3 groups. The first group is treated by brushing their teeth right after eating bread, the second and third group is treated 15 and 30 minutes after eating bread. The hypothesis uses Kruskal Wallis hypothesis continued by Mann Whitney test, strikethrough. The study reveals that the group brushed their teeth right after eating bread shows low category of saliva buffering is that 55% meanwhile those who brushed their teeth 15 and 30 minutes after eating bread exhibits the result as much as 65% and 25 % Thus the last group is included to those who have a medium risk of suffering from dental carries. The statistics of Kruskal Wallis test within the confidence level of 95% shows that there is an influence of teeth brushing time over the saliva buffering capacity with p<0,001. Mann Whitney test shows that the time of teeth brushing within 15 minutes after eating is better than the group who brush their teeth 30 minutes after eating.

  17. Plasma and saliva concentrations of abacavir, tenofovir, darunavir, and raltegravir in HIV-1-infected patients
.

    Science.gov (United States)

    Yamada, Eiko; Takagi, Ritsuo; Tanabe, Yoshinari; Fujiwara, Hiroshi; Hasegawa, Naoki; Kato, Shingo

    2017-07-01

    We studied the relationships between plasma and saliva concentrations of antiretroviral drugs to explore whether saliva can be used for therapeutic drug monitoring (TDM). Abacavir (ABC), tenofovir (TFV), darunavir (DRV), and raltegravir (RAL) in plasma and saliva from 30 HIV-1-infected patients were quantified using liquid chromatography-tandem mass spectrometry. Mean saliva-to-plasma concentration ratios were 0.623 (ABC), 0.024 (TFV), 0.065 (DRV), and 0.0135 (RAL), which agree with the plasma protein binding rates except TFV. Significant correlations were evident between saliva and plasma concentrations of ABC, DRV, and RAL. This study suggests that plasma concentrations of ABC, DRV, and RAL can be estimated from their saliva concentrations and that the saliva concentration of some antiretroviral drugs reflects the unbound drug concentration in plasma.
.

  18. Isolation of a protein-containing cell surface component from Streptococcus sanguis which affects its adherence to saliva-coated hydroxyapatite.

    OpenAIRE

    Liljemark, W. F.; Bloomquist, C G

    1981-01-01

    The isolation and partial characterization of a protein-containing cell surface component from Streptococcus sanguis which blocks the adherence of this microbe to saliva-coated hydroxyapatite are described. Several methods of extraction were attempted. Sonication of whole cells and cell walls proved to be the most successful and yielded biologically active adherence-blocking components. The adherence-blocking ability of these components was effective in intraspecies blocking experiments. The ...

  19. HUBUNGAN FAKTOR RESIKO KARIES DALAM SALIVA DENGAN INDEKS DMF-T PADA PENDERITA DM TIPE II DI RSCM SUB BAGIAN ENDOKRIN (Laporan Penelitian

    Directory of Open Access Journals (Sweden)

    Buddiwati Punta

    2015-07-01

    Full Text Available Dental caries is a multifactorial disease. The most important factors in the development of caries is saliva. Reduced salivary secretion in patients with type 2 diabetes melitus to the occurrence of caries have yielded controversial results. The aim of the study was to find out whether the risk factors of the saliva i.e. saliva secretion rate, buffer capacity, salivary S. mutans, salivary Lactobacilli, alone or in combination, could be used for prediction of caries activity. Thirty patients with type 2 diabetes mellitus and age range 46-73 participated. Diabetic status was determined by fasting plasma glucose is >126mg/dl, 2-hour plasma glucose is >140mg/dl. Decayed, missing, and filled teeth indices were determined by means of clinical examination. Stimulated saliva with parafin was measured for flow rate and buffer capacity, level of S. mutans & Lactobacilli were analyzed with dentobuff and dentocult. There was no decrease in salivary secretion in patients with type 2 diabetes melitus. Significant corellation were found between buffer capacity, and combination of the S. mutans & Lactobacilli counts in caries activity.

  20. Bacterial composition in whole saliva from patients with severe hyposalivation

    DEFF Research Database (Denmark)

    Belstrøm, Daniel; Holmstrup, Palle; Fiehn, Nils-Erik

    2016-01-01

    OBJECTIVE: The purpose of this study was to compare the microbiota of stimulated whole saliva samples from patients with severe hyposalivation to samples from individuals with normal whole saliva flow rates. It was hypothesized that the two groups differ with regard to salivary bacterial profiles....... METHODS: This cross-sectional study included 36 participants (24 females and 12 males, mean age 58.5 years) with severe hyposalivation and 36 gender-, age- and geographically-matched participants with normal salivary secretion from the Danish Health Examination Survey (DANHES). The microbiota...... with severe hyposalivation do not differ from those of individuals with normal salivary secretion, when there are virtually no untreated active caries lesions present in the oral cavity. This article is protected by copyright. All rights reserved....

  1. Comparison between three different saliva substitutes in patients with hyposalivation.

    Science.gov (United States)

    Skrinjar, Ivana; Vucicevic Boras, Vanja; Bakale, Iva; Andabak Rogulj, Ana; Brailo, Vlaho; Vidovic Juras, Danica; Alajbeg, Ivan; Vrdoljak, Danko Velimir

    2015-04-01

    The aim of the present study was to compare the efficiency of oral spray based on thermal spring water (Buccotherm®) versus commercial saliva substitute (Xeros®) and marshmallow root on the quality of life in patients with hyposalivation. A total of 60 patients with unstimulated salivary flow rate hyposalivation. Intensity of dry mouth was lower after the applied therapy whatever substitute patients used. We recommend the use of all three saliva substitutes for decreasing the intensity of dry mouth symptoms as well as improvement in the quality of life. Although all tested agents showed beneficial effect in alleviating hyposalivation symptoms, it seems that Buccotherm® was superior to Xeros® and marshmallow root.

  2. Corrosion Behavior of Titanium in Artificial Saliva by Lactic Acid

    Directory of Open Access Journals (Sweden)

    Qing Qu

    2014-07-01

    Full Text Available As one of the main products produced by oral microorganisms, the role of lactic acid in the corrosion of titanium is very important. In this study, the corrosion behavior of titanium in artificial saliva with and without lactic acid were investigated by open-circuit potentials (OCPs, polarization curves and electrochemical impedance spectroscopy (EIS. OCP firstly increased with the amount of lactic acid from 0 to 3.2 g/L and then tended to decrease from 3.2 to 5.0 g/L. The corrosion of titanium was distinctly affected by lactic acid, and the corrosion rate increased with increasing the amount of lactic acid. At each concentration of lactic acid, the corrosion rate clearly increased with increasing the immersing time. Results of scanning electron microscopy (SEM also indicated that lactic acid accelerated the pitting corrosion in artificial saliva. A probable mechanism was also proposed to explain the experimental results.

  3. Investigation of saliva of patients with periodontal disease using NAA

    Energy Technology Data Exchange (ETDEWEB)

    Zamboni, C. B.; Metairon, S.; Medeiros, I. M. M. A. [Instituto de Pesquisas Energeticas e Nucleares, IPEN - CNEN/SP Av. Professor Lineu Prestes 2242- 05508-000 Sao Paulo, SP (Brazil); Lewgoy, H. R. [Universidade Anhanguera Bandeirante, UNIBAN R. Maria Candida, 1813, Bloco G / 6o andar - 02071-013 Sao Paulo, SP (Brazil)

    2013-05-06

    In this study the non-stimulated whole saliva of 26 healthy subjects (mean age 33.9 {+-} 11.0 years, range: 26 to 49 years) and 11 patients with periodontal disease (mean age 41.7 {+-} 11.5 years; range 29 to 55 years) was investigated using Neutron Activation Analysis (NAA) technique. The samples were obtained from donors at Sao Paulo city (Brazil). The analyses were performed in the nuclear reactor IEA-R1 (3.5-4.5MW, pool type) at IPEN/CNEN-SP (Brazil). Considerable changes in Ca and S saliva's level were identified in patients with periodontal disease suggesting they can be used as monitors of periodontal diseases.

  4. Investigation of saliva of patients with periodontal disease using NAA

    Science.gov (United States)

    Zamboni, C. B.; Metairon, S.; Medeiros, I. M. M. A.; Lewgoy, H. R.

    2013-05-01

    In this study the non-stimulated whole saliva of 26 healthy subjects (mean age 33.9 ± 11.0 years, range: 26 to 49 years) and 11 patients with periodontal disease (mean age 41.7 ± 11.5 years; range 29 to 55 years) was investigated using Neutron Activation Analysis (NAA) technique. The samples were obtained from donors at São Paulo city (Brazil). The analyses were performed in the nuclear reactor IEA-R1 (3.5-4.5MW, pool type) at IPEN/CNEN-SP (Brazil). Considerable changes in Ca and S saliva's level were identified in patients with periodontal disease suggesting they can be used as monitors of periodontal diseases.

  5. Simultaneous analysis of micro-RNA and DNA for determining the body fluid origin of DNA profiles.

    Science.gov (United States)

    van der Meer, Donny; Uchimoto, Mari L; Williams, Graham

    2013-07-01

    Micro-RNAs (miRNAs) can be specifically expressed in forensically relevant body fluids such as blood or saliva. The aim of the study was to develop a simultaneous extraction and analysis protocol that allows for the acquisition of a DNA profile and the identity of the body fluid using a single process. DNA and micro-RNA were extracted from blood and saliva before undergoing a cDNA synthesis step by using stem-loop reverse transcription PCR. The resulting extracts containing DNA and cDNA synthesized from body fluid-specific miRNA markers then underwent standard STR analysis using a modified ABI AmpFℓSTR(®) NGM SElect™ kit. In all samples, a full DNA profile was obtained along with additional peaks corresponding to the miRNA marker targeted. In all cases, blood samples profiled exhibited a peak indicating the presence of the blood-specific miRNA marker and the saliva sample profiled exhibited a peak indicating the presence of the saliva-specific miRNA marker. © 2013 American Academy of Forensic Sciences.

  6. 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.

  7. Systematic comparison of the human saliva and plasma proteomes

    OpenAIRE

    Yan, Weihong; Apweiler, Rolf; Balgley, Brian M.; Boontheung, Pinmanee; Bundy, Jonathan L.; Cargile, Benjamin J.; Cole, Steve; Fang, Xueping; Gonzalez-Begne, Mireya; Griffin, Timothy J.; Hagen, Fred; Hu, Shen; Wolinsky, Lawrence E.; Lee, Cheng S.; Malamud, Daniel

    2009-01-01

    The proteome of human salivary fluid has the potential to open new doors for disease biomarker discovery. A recent study to comprehensively identify and catalog the human ductal salivary proteome led to the compilation of 1166 proteins. The protein complexity of both saliva and plasma is large, suggesting that a comparison of these two proteomes will provide valuable insight into their physiological significance and an understanding of the unique and overlapping disease diagnostic potential t...

  8. Noninvasive glucose monitoring using saliva nano-biosensor

    OpenAIRE

    Zhang, Wenjun; Du, Yunqing; Wang, Ming L.

    2015-01-01

    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...

  9. Exploiting leech saliva to treat osteoarthritis: A provocative perspective

    Directory of Open Access Journals (Sweden)

    Edwin L. Cooper

    2017-07-01

    Full Text Available Plant and animal-derived products are crucial components in complementary and alternative medicine. Although modern medicine has provided numerous innovations and advancements, these often fail to reveal new and dependable, inexpensive treatments nor real cures that are relatively free of adverse side effects. We present evidence that hirudotherapy, which utilizes leeches, improves certain diseases, including osteoarthritis. Osteoarthritis, a disease in joints, could benefit from use of medicinal peptides found in leech saliva, components of its immune system.

  10. The effect of saliva on the fate of nanoparticles.

    Science.gov (United States)

    Teubl, Birgit J; Stojkovic, Biljana; Docter, Dominic; Pritz, Elisabeth; Leitinger, Gerd; Poberaj, Igor; Prassl, Ruth; Stauber, Roland H; Fröhlich, Eleonore; Khinast, Johannes G; Roblegg, Eva

    2017-07-09

    The design of nanocarriers for local drug administration to the lining mucosa requires a sound knowledge of how nanoparticles (NPs) interact with saliva. This contact determines whether NPs agglomerate and become immobile due to size- and interaction-filtering effects or adsorb on the cell surface and are internalized by epithelial cells. The aim of this study was to examine the behavior of NPs in saliva considering physicochemical NP properties. The salivary pore-size distribution was determined, and the viscosity of the fluid inside of the pores was studied with optical tweezers. Distinct functionalized NPs (20 and 200 nm) were dispersed in saliva and salivary buffers and characterized, and surface-bound MUC5B and MUC7 were analyzed by 1D electrophoresis and immunoblotting. NP mobility was recorded, and cellular uptake studies were performed with TR146 cells. The mode diameter of the salivary mesh pores is 0.7 μm with a peak width of 1.9 μm, and pores are filled with a low-viscosity fluid. The physicochemical properties of the NPs affected the colloidal stability and mobility: compared with non-functionalized particles, which did not agglomerate and showed a cellular uptake rate of 2.8%, functionalized particles were immobilized, which was correlated with agglomeration and increased binding to mucins. The present study showed that the salivary microstructure facilitates NP adsorption. However, NP size and surface functionalization determine the colloidal stability and cellular interactions. The sound knowledge of NP interactions with saliva enables the improvement of current treatment strategies for inflammatory oral diseases.

  11. The Food Preferences of the Blow Fly Lucilia cuprina Offered Human Blood, Semen and Saliva, and Various Nonhuman Foods Sources.

    Science.gov (United States)

    Durdle, Annalisa; Mitchell, Robert J; van Oorschot, Roland A H

    2016-01-01

    As human DNA profiles can be obtained from blow fly artifacts, this study aimed to establish the feeding preferences of Lucilia cuprina (Wiedemann) blow flies when offered human biological fluids and nonhuman food sources. One-day-old and 3-day-old blow flies of both sexes were simultaneously offered human blood, semen and saliva, pet food, canned tuna and honey, and the number and length of visits documented over 6 h. One-day-old flies visited pet food and honey most often, but stayed longest on honey and semen. Three-day-old flies visited semen and pet food most often, and stayed longest on these food sources. Blood and saliva were the least preferred options for all flies. Overall, flies preferred dry blood and semen to the wet forms. These findings demonstrate that even when other food sources are available, flies at a crime scene may feed on human biological fluids if present, potentially transferring human DNA. © 2015 American Academy of Forensic Sciences.

  12. Saliva: a powerful diagnostic tool for minimal intervention dentistry.

    Science.gov (United States)

    Ranganath, L M; Shet, R G K; Rajesh, A G

    2012-03-01

    Saliva plays a vital role in oral health as patients strive to maintain a healthy dentition throughout their lives. It is natures primary defense mechanism for the oral environment, and is particularly important for protecting exposed tooth surfaces. While internal protection for dentin comes from odontoblasts and the dental pulp, the body's external protection for enamel comes from saliva. The noninvasive nature of salivary testing has made it an effective alternative to blood and urine testing and home testing kits have made it possible for people to monitor their own health using this diagnostic medium. This paper presents what saliva can reveal about general and oral health as well as highlights the current use and potential clinical and research applications, of diagnostics based on oral fluids. Early detection always minimizes the need for more invasive treatment. It prevents oral health disease at an early stage and provides a good oral health in rejuvenated state. If you stick and follow regular professional care, prevention maintenance appointments, prevention counseling, good home care and oral hygiene, diet habits you will be free from oral health illness and you can experience the harmonious and rejuvenated state of good oral health.

  13. Validation of a New Saliva Test for Helicobacter pylori Infection

    Directory of Open Access Journals (Sweden)

    OF Bathe

    1996-01-01

    Full Text Available The purpose of this study was to evaluate a recently introduced saliva test measuring immunoglobulin (Ig G antibodies to Helicobacter pylori by enzyme-linked immunosorbent assay (ELISA. ELISA has previously been validated against IgG serological tests; however, it is not considered the definitive test for H pylori infection. Using endoscopic antral biopsies as the ’gold standard’ for comparison, the saliva test was validated on 70 patients with upper gastrointestinal symptoms admitted to St Paul’s Hospital Gastroenterology Clinic for gastroduodenoscopy. Thirty-five patients (50% had histological evidence of gastritis and, by using acridine orange stain, the bacterium was visualized in 25 patients (71%. A biopsy was considered positive when the bacterium was visualized. The saliva test was determined to be 80% sensitive and 80% specific for H pylori infection when the cut-off for a positive test was 0.3 ELISA U/mL. Positive and negative predictive values were 69% and 88%, respectively. More study is required to assess the clinical utility of the test.

  14. Predominant presence of Streptococcus anginosus in the saliva of alcoholics.

    Science.gov (United States)

    Morita, E; Narikiyo, M; Yokoyama, A; Yano, A; Kamoi, K; Yoshikawa, E; Yamaguchi, T; Igaki, H; Tachimori, Y; Kato, H; Saito, D; Hanada, N; Sasaki, H

    2005-12-01

    Chronic alcohol consumption is known to be a major risk factor for cancers of the upper aerodigestive tract. The incidence of esophageal cancer (4.4%) in alcoholics is reported to be much higher than that in the Japanese population as a whole (0.0001%). This suggests the presence of specific factors in chronic alcohol consumption-related carcinogenesis. Recently, data showing a significant correlation between Streptococcus anginosus and carcinogenesis in the upper aerodigestive tract have been reported. In this study, the ratio of S. anginosus to oral bacteria in the saliva of 38 alcoholic patients was investigated to determine if there is an association between alcoholic patients and S. anginosus infection. The level of S. anginosus in the saliva from 22 healthy people, 41 esophageal cancer patients, 32 gastritis patients, and 24 periodontitis patients was also investigated and compared to the level in alcoholic patients. In the saliva from esophageal cancer patients, the level of S. anginosus was not significantly different from that of healthy people. The levels of S. anginosus in periodontitis and gastritis patients were also similar. In alcoholics, however, there was an extremely high level of S. anginosus, suggesting that they, rather than healthy people and general esophageal cancer patients, have a high risk for S. anginosus infection.

  15. Detection of sulphate-reducing bacteria in human saliva.

    Science.gov (United States)

    Heggendorn, Fabiano Luiz; Gonçalves, Lucio Souza; Dias, Eliane Pedra; Silva Junior, Arley; Galvão, Mariana Machado; Lutterbach, Márcia T S

    2013-11-01

    The aim of the current study was to investigate the presence of sulphate-reducing bacteria (SRB) in human saliva and correlate with oral and systemic conditions. Saliva samples were collected from 118 patients and inoculated in 2 ml of modified Postgate's E medium culture. After 28 days of incubation at 30°C the presence of SRB was identified by the production of sulphide. Of 118 saliva samples collected, 35 were positive for the presence of SRB. Three positive samples were randomly chosen to identify the species of SRB by PCR and sequenced. The three selected samples were identified as Desulfovibrio fairfieldensis, Desulfovibrio desulfuricans and Raoultella ornithinolytica. Gastritis (14.4%) was the most prevalent systemic disease, followed by diabetes (3.4%), while periodontitis (11%) and traumatic fibroma (4.2%) were the oral manifestations most frequently found. A bivariate analysis was performed to examine for the presence of SRB and the most prevalent systemic and oral manifestations. Only periodontitis showed a statistically significant association (p = 0.0003). The results showed SRB can be found in oral microbiota of healthy patients. Regarding the several conditions studied, there was a higher prevalence of SRB in patients with gastritis and patients with periodontal disease, with a possible correlation between the presence of SRB in the oral microbiota and periodontal disease.

  16. Characterization of the Activity and Stability of Amylase from Saliva and Detergent: Laboratory Practicals for Studying the Activity and Stability of Amylase from Saliva and Various Commercial Detergents

    Science.gov (United States)

    Valls, Cristina; Rojas, Cristina; Pujadas, Gerard; Garcia-Vallve, Santi; Mulero, Miquel

    2012-01-01

    This article presents two integrated laboratory exercises intended to show students the role of [alpha]-amylases (AAMYs) in saliva and detergents. These laboratory practicals are based on the determination of the enzymatic activity of amylase from saliva and different detergents using the Phadebas test (quantitative) and the Lugol test…

  17. 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.

  18. Aphid Gel Saliva: Sheath Structure, Protein Composition and Secretory Dependence on Stylet-Tip Milieu

    Science.gov (United States)

    Will, Torsten; Steckbauer, Kathrin; Hardt, Martin; van Bel, Aart J. E.

    2012-01-01

    In order to separate and analyze saliva types secreted during stylet propagation and feeding, aphids were fed on artificial diets. Gel saliva was deposited as chains of droplets onto Parafilm membranes covering the diets into which watery saliva was secreted. Saliva compounds collected from the diet fluid were separated by SDS-PAGE, while non-soluble gel saliva deposits were processed in a novel manner prior to protein separation by SDS-PAGE. Soluble (watery saliva) and non-soluble (gel saliva) protein fractions were significantly different. To test the effect of the stylet milieu on saliva secretion, aphids were fed on various diets. Hardening of gel saliva is strongly oxygen-dependent, probably owing to formation of sulfide bridges by oxidation of sulphydryl groups. Surface texture of gel saliva deposits is less pronounced under low-oxygen conditions and disappears in dithiothreitol containing diet. Using diets mimicking sieve-element sap and cell-wall fluid respectively showed that the soluble protein fraction was almost exclusively secreted in sieve elements while non-soluble fraction was preferentially secreted at cell wall conditions. This indicates that aphids are able to adapt salivary secretion in dependence of the stylet milieu. PMID:23056521

  19. Antioxidant capacity of human saliva and periodontal screening assessment in healthy adults.

    Science.gov (United States)

    Tartaglia, Gianluca Martino; Gagliano, Nicoletta; Zarbin, Luca; Tolomeo, Giorgia; Sforza, Chiarella

    2017-06-01

    Saliva plays a pivotal role as an antioxidant system, and saliva antioxidant levels are reduced in patients with periodontal disease. Recently, a biochemical test able to determine saliva antioxidant levels was proposed as predictive for oral cavity diseases, but it was not clinically tested. In this preliminary study, we evaluated the relationships between Periodontal Screening and Recordings characteristics of patients and saliva antioxidant levels measures. Thirty-nine patients (12 men, 27 women; mean age, 46 years, SD 17) attending the dental hygiene unit of a Private Clinic underwent a Periodontal Screening and Recordings examination and a saliva antioxidant levels measurement using a biochemical commercial test. The results of the clinical periodontal examination were compared to those obtained by the saliva test. Approximately 70% of patients showed a low saliva antioxidant levels value, while the other patients had Optimal/Normal values. Thirteen patients (33%) resulted positive to Periodontal Screening and Recordings test. Using Periodontal Screening and Recordings values as gold standard, the saliva antioxidant levels test correctly classified 52.6% of patients; sensitivity was 84.6%, specificity was 36%. The saliva antioxidant levels test had a good sensitivity when compared to the gold standard; this finding corroborates the hypothesis that alterations of the oral antioxidant levels are related to periodontal disease. The reduced specificity shows that saliva antioxidant levels test could detect alterations predisposing to periodontal disease before clinically evident aspects. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Breastmilk-Saliva Interactions Boost Innate Immunity by Regulating the Oral Microbiome in Early Infancy

    Science.gov (United States)

    Al-Shehri, Saad S.; Knox, Christine L.; Liley, Helen G.; Cowley, David M.; Wright, John R.; Henman, Michael G.; Hewavitharana, Amitha K.; Charles, Bruce G.; Shaw, Paul N.; Sweeney, Emma L.; Duley, John A.

    2015-01-01

    Introduction Xanthine oxidase (XO) is distributed in mammals largely in the liver and small intestine, but also is highly active in milk where it generates hydrogen peroxide (H2O2). Adult human saliva is low in hypoxanthine and xanthine, the substrates of XO, and high in the lactoperoxidase substrate thiocyanate, but saliva of neonates has not been examined. Results Median concentrations of hypoxanthine and xanthine in neonatal saliva (27 and 19 μM respectively) were ten-fold higher than in adult saliva (2.1 and 1.7 μM). Fresh breastmilk contained 27.3±12.2 μM H2O2 but mixing baby saliva with breastmilk additionally generated >40 μM H2O2, sufficient to inhibit growth of the opportunistic pathogens Staphylococcus aureus and Salmonella spp. Oral peroxidase activity in neonatal saliva was variable but low (median 7 U/L, range 2–449) compared to adults (620 U/L, 48–1348), while peroxidase substrate thiocyanate in neonatal saliva was surprisingly high. Baby but not adult saliva also contained nucleosides and nucleobases that encouraged growth of the commensal bacteria Lactobacillus, but inhibited opportunistic pathogens; these nucleosides/bases may also promote growth of immature gut cells. Transition from neonatal to adult saliva pattern occurred during the weaning period. A survey of saliva from domesticated mammals revealed wide variation in nucleoside/base patterns. Discussion and Conclusion During breast-feeding, baby saliva reacts with breastmilk to produce reactive oxygen species, while simultaneously providing growth-promoting nucleotide precursors. Milk thus plays more than a simply nutritional role in mammals, interacting with infant saliva to produce a potent combination of stimulatory and inhibitory metabolites that regulate early oral–and hence gut–microbiota. Consequently, milk-saliva mixing appears to represent unique biochemical synergism which boosts early innate immunity. PMID:26325665

  1. Saliva fluoride before and during 3 years of supervised use of fluoride toothpaste.

    Science.gov (United States)

    Richards, A; Machiulskiene, V; Nyvad, B; Baelum, V

    2013-12-01

    The purpose of the study was to examine pre-brushing saliva fluoride concentrations before and during a large, 3-year, prospective toothpaste study on the effect of post-brushing rinsing on dental caries. The aims were to study saliva fluoride over time and the effect of rinsing on saliva fluoride and to relate saliva fluoride to caries increments and accumulation of plaque. Saliva samples (baseline and 1, 2, and 3 years) were collected from 11-year-old children attending two schools (A and B) in Kaunas, Lithuania, who refrained from brushing the evening and morning before saliva collection. Numbers of saliva samples collected varied from 264 at baseline to 188 at the 3-year follow-up. Children in school A rinsed with water after daily brushing, while children in school B did not rinse. Total caries and visible plaque were registered at baseline and after 3 years. Mean saliva fluoride concentrations at baseline and after 1, 2, and 3 years from school A (rinsing) were 0.014, 0.026, 0.029, and 0.034 ppm and from school B (no rinsing) were 0.013, 0.028, 0.031, and 0.031 ppm, respectively. Increases in saliva fluoride from baseline were significant (Wilcoxon's test, p Saliva fluoride did not increase beyond year 1 and did at no time point differ between schools. Reductions in numbers of tooth surfaces with dental plaque were significantly positively related to the number of caries reversals over the 3 years. Background saliva fluoride concentration is increased by brushing at least once daily on schooldays, does not increase further over 3 years, and is not affected by rinsing after brushing. Continuous use of fluoride toothpaste produces ambient saliva fluoride levels similar to saliva fluoride in areas with fluoridated water.

  2. Breastmilk-Saliva Interactions Boost Innate Immunity by Regulating the Oral Microbiome in Early Infancy.

    Directory of Open Access Journals (Sweden)

    Saad S Al-Shehri

    Full Text Available Xanthine oxidase (XO is distributed in mammals largely in the liver and small intestine, but also is highly active in milk where it generates hydrogen peroxide (H2O2. Adult human saliva is low in hypoxanthine and xanthine, the substrates of XO, and high in the lactoperoxidase substrate thiocyanate, but saliva of neonates has not been examined.Median concentrations of hypoxanthine and xanthine in neonatal saliva (27 and 19 μM respectively were ten-fold higher than in adult saliva (2.1 and 1.7 μM. Fresh breastmilk contained 27.3 ± 12.2 μM H2O2 but mixing baby saliva with breastmilk additionally generated >40 μM H2O2, sufficient to inhibit growth of the opportunistic pathogens Staphylococcus aureus and Salmonella spp. Oral peroxidase activity in neonatal saliva was variable but low (median 7 U/L, range 2-449 compared to adults (620 U/L, 48-1348, while peroxidase substrate thiocyanate in neonatal saliva was surprisingly high. Baby but not adult saliva also contained nucleosides and nucleobases that encouraged growth of the commensal bacteria Lactobacillus, but inhibited opportunistic pathogens; these nucleosides/bases may also promote growth of immature gut cells. Transition from neonatal to adult saliva pattern occurred during the weaning period. A survey of saliva from domesticated mammals revealed wide variation in nucleoside/base patterns.During breast-feeding, baby saliva reacts with breastmilk to produce reactive oxygen species, while simultaneously providing growth-promoting nucleotide precursors. Milk thus plays more than a simply nutritional role in mammals, interacting with infant saliva to produce a potent combination of stimulatory and inhibitory metabolites that regulate early oral-and hence gut-microbiota. Consequently, milk-saliva mixing appears to represent unique biochemical synergism which boosts early innate immunity.

  3. Effective lactation yield

    NARCIS (Netherlands)

    Kok, Akke; Middelaar, van C.E.; Engel, B.; Knegsel, van A.T.M.; Hogeveen, H.; Kemp, B.; Boer, de I.J.M.

    2016-01-01

    To compare milk yields between cows or management strategies, lactations are traditionally standardized to 305-d yields. The 305-d yield, however, gives no insight into the combined effect of additional milk yield before calving, decreased milk yield after calving, and a possible shorter calving

  4. Presence of Lactobacillus reuteri in saliva coincide with higher salivary IgA in young adults after intake of probiotic lozenges.

    Science.gov (United States)

    Braathen, G; Ingildsen, V; Twetman, S; Ericson, D; Jørgensen, M R

    2017-02-07

    The aim of this study was to compare the concentration of salivary immunoglobulin A (IgA) and the selected interleukins (IL)-1β, IL-6, IL-8 and IL-10 in young individuals with presence and non-presence of Lactobacillus reuteri in saliva after a three-week intervention with probiotic lozenges. The study group consisted of 47 healthy individuals aged 18-32 years with no clinical signs of oral inflammation. In a randomised, double-blind, placebo-controlled, cross-over trial participants ingested two lozenges per day containing two strains of the probiotic bacterium L. reuteri or placebo lozenges. The intervention and wash-out periods were three weeks. Stimulated and unstimulated whole saliva was collected at baseline and immediately after termination of the intervention periods. The samples were analysed for total protein, salivary IgA and selected cytokines. In this extended analysis, data were collected by analysing baseline and follow-up saliva samples related to ingestion of the probiotic lozenges for the presence of L. reuteri through DNA-extraction, PCR-amplification and gel-electrophoresis. At baseline, 27% of the individuals displayed presence of L. reuteri and 42% were positive immediately after the three-week probiotic intervention. Individuals with presence of L. reuteri in saliva had significantly higher (Psaliva coincided with higher concentrations of salivary IgA and %IgA/protein in stimulated whole saliva after the three-week daily intake of probiotic lozenges. Our findings suggest that monitoring the presence of probiotic candidates in the oral environment is important to interpret and understand their possible immune-modulating role in maintaining oral health.

  5. Application of agglomerative clustering for analyzing phylogenetically on bacterium of saliva

    Science.gov (United States)

    Bustamam, A.; Fitria, I.; Umam, K.

    2017-07-01

    Analyzing population of Streptococcus bacteria is important since these species can cause dental caries, periodontal, halitosis (bad breath) and more problems. This paper will discuss the phylogenetically relation between the bacterium Streptococcus in saliva using a phylogenetic tree of agglomerative clustering methods. Starting with the bacterium Streptococcus DNA sequence obtained from the GenBank, then performed characteristic extraction of DNA sequences. The characteristic extraction result is matrix form, then performed normalization using min-max normalization and calculate genetic distance using Manhattan distance. Agglomerative clustering technique consisting of single linkage, complete linkage and average linkage. In this agglomerative algorithm number of group is started with the number of individual species. The most similar species is grouped until the similarity decreases and then formed a single group. Results of grouping is a phylogenetic tree and branches that join an established level of distance, that the smaller the distance the more the similarity of the larger species implementation is using R, an open source program.

  6. A novel two-component system of Streptococcus sanguinis affecting functions associated with viability in saliva and biofilm formation.

    Science.gov (United States)

    Camargo, Tarsila M; Stipp, Rafael N; Alves, Lívia A; Harth-Chu, Erika N; Höfling, José F; Mattos-Graner, Renata O

    2018-01-16

    Streptococcus sanguinis is a pioneer species of teeth and a common opportunistic pathogen of infective endocarditis. In this study, we identified a two-component system, SptRS Ss , affecting S. sanguinis survival in saliva and biofilm formation. Isogenic mutants of sptRSs (SKsptR) and sptSSs (SKsptS) showed reduced cell counts in ex vivo assays of viability in saliva, compared to parent strain SK36 and complemented mutants. Reduced counts of the mutants in saliva were associated with reduced growth rates in poor nutrient medium (RPMI) and increased susceptibility to deposition of C3b and MAC of the complement system, a defense component of saliva and serum. Conversely, sptR/SSs mutants showed increased biofilm formation associated with higher production of H2O2 and extracellular DNA. RT-qPCR comparisons of strains indicated a global role of SptRS Ss in repressing genes for H2O2 production (2.5 to 15-fold up-regulation of spxB, spxR, vicR, tpk and ackA in sptR/SSs mutants), biofilm formation and/or evasion to host immunity (2.1 to 11.4-fold up-regulation of srtA, pcsB, cwdP, iga, nt5e). Compatible with SptR Ss homology with AraC-type regulators, duplicate to multiple conserved repeats were identified in 1,000 bp regulatory regions of downstream genes, suggesting that SptR Ss regulates transcription by DNA looping. Significant transcriptional changes in the regulatory genes vicR, spxR, comE, comX, and mecA in the sptR/SSs mutants, further indicated that SptRS Ss is part of a regulatory network which coordinates cell wall homeostasis, H2O2 production, and competence. This study reveals that SptRS Ss is involved in the regulation of crucial functions for S. sanguinis persistence in the oral cavity. Copyright © 2018 American Society for Microbiology.

  7. Glucose Uptake by Streptococcus mutans, Streptococcus mitis, and Actinomyces viscosus in the Presence of Human Saliva

    Science.gov (United States)

    Germaine, Greg, R.; Tellefson, Lois M.

    1982-01-01

    Glucose uptake was examined by using whole-cell suspensions of Streptococcus mutans (strains BHT, Ingbritt, and GS-5), Streptococcus mitis (strains 9811 and 72×41), and Actinomyces viscosus (strains T6 and WVU626) incubated for up to 90 min in 0 to 82% (vol/vol) human whole salivary supernatant. Glucose uptake by the S. mutans strains was completely inhibited at all saliva concentrations. Dithiothreitol (DTT), present during saliva incubation, prevented saliva inhibition. Glucose uptake was also restored when saliva-inhibited cells were subsequently exposed to DTT. The inclusion of catalase in the saliva incubation mixtures resulted in protection equal to that obtained with DTT. The S. mitis strains were also inhibited by saliva but to a far lesser extent that S. mutans. DTT and catalase also protected S. mitis from saliva inhibition. Both A. viscosus strains were completely refractory to saliva inhibition of glucose uptake. Based on (i) the sensitivity of the catalase-negative streptococci and the resistance of catalase-positive actinomyces to saliva inhibition and (ii) the equal and complete protection to saliva inhibition afforded by DTT and catalase, we conclude that the lactoperoxidase-SCN−-H2O2 system in saliva was the only antibacterial system expressed under our experimental conditions. The relative resistance of S. mitis 9811 (compared with S. mutans BHT) to saliva inhibition was shown not to result from poor H2O2 production in either glucose-supplemented buffer or saliva solutions. S. mitis produced inhibitory quantities of H2O2 that equaled or exceeded S. mutans H2O2 accumulation. It is suggested that S. mitis might possess a greater ability to repair lactoperoxidase-mediated damage than does S. mutans. Every organism studied exhibited a saliva concentration-dependent, cell growth-independent stimulation of glucose uptake after 60 to 90 min of incubation. The A. viscosus and S. mitis strains showed saliva stimulation (or stabilization) of glucose

  8. 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

  9. Oral contraceptive use and saliva diurnal pattern of metabolic steroid hormones in young healthy women.

    Science.gov (United States)

    Vibarel-Rebot, N; Rieth, N; Lasne, F; Jaffré, C; Collomp, K

    2015-03-01

    The impact of oral contraceptives (OCs) on the saliva diurnal pattern of metabolic steroid hormones remained unknown. Saliva samples were taken from young healthy women (11 OC users, 10 non-OC users) to analyze cortisol, dehydroepiandrosterone (DHEA) and testosterone 4 times (days 1, 8, 15 and 22) over one menstrual cycle. OC use decreased saliva testosterone concentrations (ppattern. The clinical relevance requires further study. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Saliva versus Plasma for Pharmacokinetic and Pharmacodynamic Studies of Fentanyl in Patients with Cancer.

    Science.gov (United States)

    Bista, Sudeep R; Haywood, Alison; Norris, Ross; Good, Phillip; Tapuni, Angela; Lobb, Michael; Hardy, Janet

    2015-11-01

    Fentanyl is widely used to relieve cancer pain. However there is great interpatient variation in the dose required to relieve pain and little knowledge about the pharmacokinetic and pharmacodynamic (PK/PD) relationship of fentanyl and pain control. Patients with cancer are fragile and there is reluctance on the part of health professionals to take multiple plasma samples for PK/PD studies. The relationship between plasma and saliva fentanyl concentrations was investigated to determine whether saliva could be a valid substitute for plasma in PK/PD studies. One hundred sixty-three paired plasma and saliva samples were collected from 56 patients prescribed transdermal fentanyl (Durogesic, Janssen-Cilag Pty Limited, NSW, Australia) at varying doses (12-200 µg/h). Pain scores were recorded at the time of sampling. Fentanyl and norfentanyl concentrations in plasma and saliva were quantified using HPLC-MS/MS. Saliva concentrations of fentanyl (mean = 4.84 μg/L) were much higher than paired plasma concentrations of fentanyl (mean = 0.877 μg/L). Both plasma and saliva mean concentrations of fentanyl were well correlated with dose with considerable interpatient variation at each dose. The relationship between fentanyl and norfentanyl concentrations was poor in both plasma and saliva. No correlation was observed between fentanyl concentration in plasma and saliva (r(2) = 0.3743) or free fentanyl in plasma and total saliva concentrations (r(2) = 0.1374). Pain scores and fentanyl concentration in either of the matrices were also not correlated. No predictive correlation was observed between plasma and saliva fentanyl concentration. However the detection of higher fentanyl concentrations in saliva than plasma, with a good correlation to dose, may allow saliva to be used as an alternative to plasma in PK/PD studies of fentanyl in patients with cancer. Copyright © 2015 Elsevier HS Journals, Inc. All rights reserved.

  11. The Proteomes of Human Parotid and Submandibular/Sublingual Gland Salivas Collected as the Ductal Secretions

    OpenAIRE

    Denny, Paul; Hagen, Fred K.; Hardt, Markus; Liao, Lujian; Yan, Weihong; Arellanno, Martha; Bassilian, Sara; Bedi, Gurrinder S.; Boontheung, Pinmannee; Cociorva, Daniel; Delahunty, Claire M.; Denny, Trish; Dunsmore, Jason; Faull, Kym F.; Gilligan, Joyce

    2008-01-01

    Saliva is a body fluid with important functions in oral and general health. A consortium of three research groups catalogued the proteins in human saliva collected as the ductal secretions: 1166 identifications—914 in parotid and 917 in submandibular/sublingual saliva—were made. The results showed that a high proportion of proteins that are found in plasma and/or tears are also present in saliva along with unique components. The proteins identified are involved in numerous molecular processes...

  12. Glucose Uptake by Streptococcus mutans, Streptococcus mitis, and Actinomyces viscosus in the Presence of Human Saliva

    OpenAIRE

    Germaine, Greg, R.; Tellefson, Lois M.

    1982-01-01

    Glucose uptake was examined by using whole-cell suspensions of Streptococcus mutans (strains BHT, Ingbritt, and GS-5), Streptococcus mitis (strains 9811 and 72×41), and Actinomyces viscosus (strains T6 and WVU626) incubated for up to 90 min in 0 to 82% (vol/vol) human whole salivary supernatant. Glucose uptake by the S. mutans strains was completely inhibited at all saliva concentrations. Dithiothreitol (DTT), present during saliva incubation, prevented saliva inhibition. Glucose uptake was a...

  13. A tentative model for (D)-glucose turnover in human saliva.

    Science.gov (United States)

    Cetik, Sibel; Zhang, Ying; Hupkens, Emeline; Jurysta, Cedric; Malaisse, Willy J; Sener, Abdullah

    2013-10-01

    The aim of the present study is to propose a tentative model for d-glucose turnover in human saliva. The whole saliva and the saliva from parotid and submandibular/sublingual glands were collected by use of the Salivette™. The saliva glucose concentration was measured by the hexokinase method, saliva bacteria glycolysis by use of d-[5-(3)H] glucose, and the saliva ATP content by the luciferase method. The concentration of glucose amounted to 43.9±6.3 (n=29), 197.5±17.3 (n=29), 104.0±12.4 (n=27) μM in whole saliva, parotid saliva and submandibular/sublingual saliva, respectively. The rate of d-glucose utilization by oral bacteria at a physiological concentration of d-glucose in saliva (50μM) was estimated at 0.047±0.003 (n=11) nmol/min per 10(6) bacteria. Unstimulated salivary d-glucose turnover rate, as calculated from the amount of glucose secreted in saliva which comes from parotid and submandibular and sublingual glands represented 214.6±19.1%/min. In order for salivary d-glucose production to match bacterial utilization of the hexose, the total number of oral bacteria was estimated at about 2.0×10(9) bacteria, in fair agreement with previously published data. This study thus provides support for a tentative model for d-glucose turnover in human saliva. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Detection of HCV-RNA in saliva of patients with chronic hepatitis C.

    OpenAIRE

    Couzigou, P; Richard, L; Dumas, F; Schouler, L; Fleury, H

    1993-01-01

    Previous studies have provided conflicting results on the presence of hepatitis C virus-RNA in saliva. In this study, 23 (62%) of 37 patients tested positive for hepatitis C virus-RNA in saliva, using polymerase chain reaction analysis. A slightly greater proportion had a sporadic rather than a parenteral origin of chronic hepatitis C. These results provide a biological basis for saliva as a possible source of hepatitis C virus (HCV) infection, but do not necessarily imply transmission by thi...

  15. Activation of defense mechanism in wheat by polyphenol oxidase from aphid saliva.

    Science.gov (United States)

    Ma, Rui; Chen, Ju-Lian; Cheng, Deng-Fa; Sun, Jing-Rui

    2010-02-24

    The saliva of two cereal aphids, Sitobion avenae and Schizaphis graminum in third-instar nymphs, was collected after 24 h of feeding by 30 aphids, separately, on artificial diet sachets, and the salivary enzymes were determined. The result showed that polyphenol oxidase (PPO) existed in the saliva of both aphid species, and the enzymatic activities were 6.2 x 10(-3) U/g for S. avenae and 2.37 x 10(-1) U/g for S. graminum, revealing a 38-fold higher activity in the saliva of S. graminum than in the saliva of S. avenae. It was speculated that the higher PPO activity in S. graminum saliva was a contributing factor to the light yellow spot left on the feeding site of the wheat leaf by S. graminum; no such spot was left by S. avenae. After treatment of a wheat seedling with the saliva of S. avenae and S. graminum and PPO at the concentration of aphid saliva, transcript profiling data showed that aphid saliva and PPO significantly induced expression of the genes aos and fps. Because genes aos and fps encode the key enzymes in the defense signal pathways jasmonic acid and terpene signal pathways, respectively, it was deduced that PPO from aphid saliva, as the main elicitor, triggers an appropriate defense response in wheat through jasmonic acid and terpene signal pathways.

  16. [Total protein and immunoglobulin concentrations in the parotid saliva of pregnant women].

    Science.gov (United States)

    Donat, H; Tymnik, G; Bernstein, L; Knauthe, H; Kessler, L

    1977-01-01

    The content of total protein and immunglobulins in the parotid saliva and blood serum of pregnant women and healthy test persons has been determined by the biuret method and radial immunofiffusion. It was stated that total protein and IgG in the parotid saliva were higher in pregnant women than in healthy test persons, whereas the IgA-levels don't show any differences. IgM was not measurable in the parotid saliva. There was no relationship between saliva and serum immunglobulins. During the pregnancy show the parotid glands another typ of reaction than nonpregnant women.

  17. Saliva--a diagnostic window to the body, both in health and in disease.

    Science.gov (United States)

    Greabu, Maria; Battino, Maurizio; Mohora, Maria; Totan, Alexandra; Didilescu, Andreea; Spinu, Tudor; Totan, Cosmin; Miricescu, Daniela; Radulescu, Radu

    2009-01-01

    Saliva, the most available and non-invasive biofluid of the human body, permanently "bathes" the oral cavity and is trying to cope with an ever-changing milieu. The oral cavity, a very complex and unique milieu due to its dual function, is the only place in the body where the mineralized tissue is exposed to the external environment in which there are complex interactions between various surfaces: host soft and hard tissues, food, air, and microorganisms. Saliva includes a large number of inorganic and organic compounds, which act as a "mirror of the body's health." In addition to its other functions, saliva could constitute the first line of defense against oxidative stress. Due to its composition and functions, saliva could have a significant role in controlling and/or modulating oxidative damages in the oral cavity. As a diagnostic fluid, saliva offers distinctive advantages over serum. Furthermore, saliva may provide a cost-effective approach for the screening of large populations. Gland-specific saliva can be used for diagnosis of pathology specific to one of the major salivary glands. Whole saliva, however, is most frequently used for diagnosis of systemic diseases. As we enter the era of genomic medicine, sialochemistry will play an increasingly important role in the early detection, the monitoring and progression of the systemic and oral diseases. We reviewed the current data within literature and of our research concerning clinical potential of the saliva.

  18. Sample Stability and Protein Composition of Saliva: Implications for Its Use as a Diagnostic Fluid

    Directory of Open Access Journals (Sweden)

    Han Roelofsen

    2008-01-01

    Full Text Available Saliva is an easy accessible plasma ultra-filtrate. Therefore, saliva can be an attractive alternative to blood for measurement of diagnostic protein markers. Our aim was to determine stability and protein composition of saliva. Protein stability at room temperature was examined by incubating fresh whole saliva with and without inhibitors of proteases and bacterial metabolism followed by Surface Enhanced Laser Desorption/Ionization (SELDI analyses. Protein composition was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE fractionation of saliva proteins followed by digestion of excised bands and identification by liquid chromatography tandem mass spectrometry (LC-MS/MS. Results show that rapid protein degradation occurs within 30 minutes after sample collection. Degradation starts already during collection. Protease inhibitors partly prevented degradation while inhibition of bacterial metabolism did not affect degradation. Three stable degradation products of 2937 Da, 3370 Da and 4132 Da were discovered which can be used as markers to monitor sample quality. Saliva proteome analyses revealed 218 proteins of which 84 can also be found in blood plasma. Based on a comparison with seven other proteomics studies on whole saliva we identified 83 new saliva proteins. We conclude that saliva is a promising diagnostic fl uid when precautions are taken towards protein breakdown.

  19. [Analysis of the infection status of saliva Helicobacter pylori in Lanzhou].

    Science.gov (United States)

    Guo, Rui; Che, Tuanjie; Ju, Jun; Yang, Sen; He, Xiangyi; Zhang, Ying

    2014-08-01

    To determine the prevalence of saliva Helicobacter pylori in Lanzhou and investigate Helicobacter pylori-related diseases. Helicobacter pylori was detected through bacterial culture, Gram stain microscopy, and urease test from saliva samples collected from 941 residents of Lanzhou. The infection rate and growth of Helicobacter pylori among the residents were analyzed in terms of different oral health conditions, oral disease, gender, urban and rural status, and age. The rate of Helicobacter pylori-positive saliva in Lanzhou was 42.72%. The status of Helicobacter pylori infection showed significant difference among subjects with different oral hygiene and oral diseases. The rate of Helicobacter pylori-positive saliva among females was 47.89%, which was greater compared with the rate among males (38.45%, P = 0.004, chi2 = 8.492). The rate of Helicobacter pylori-positive saliva in the town was 33.99%, which was less than the rate for the villages (50.93%, P = 0.000, chi2 = 27.551). The rate of Helicobacter pylori-positive saliva among residents aged 10 to 59 showed a flat trend with no significant differences. However, the rate of Helicobacter pylori-positive saliva among residents over 60 years old showed a significant increase. No significant difference was found in the growth of saliva Helicobacter pylori (P = 0.086). The rate of Helicobacter pylori-positive saliva is related to the subjects' oral hygiene, oral disease, gender, age, and living conditions.

  20. Dental caries in young adults regarding saliva's microbiological and physical-chemical characteristics

    National Research Council Canada - National Science Library

    Martínez-Pabón, María C; Morales-Uchima, Sandra M; Martínez-Delgado, Cecilia M

    2013-01-01

    Determining the relationship between saliva's physicochemical properties, cariogenic microorganism count, facultative anaerobic and gram-negative bacteria based on caries' experience in young adults...

  1. Immunity to Lutzomyia whitmani Saliva Protects against Experimental Leishmania braziliensis Infection.

    Science.gov (United States)

    Gomes, Regis; Cavalcanti, Katrine; Teixeira, Clarissa; Carvalho, Augusto M; Mattos, Paulo S; Cristal, Juqueline R; Muniz, Aline C; Miranda, José Carlos; de Oliveira, Camila I; Barral, Aldina

    2016-11-01

    Previous works showed that immunization with saliva from Lutzomyia intermedia, a vector of Leishmania braziliensis, does not protect against experimental infection. However, L. braziliensis is also transmitted by Lutzomyia whitmani, a sand fly species closely related to Lu. intermedia. Herein we describe the immune response following immunization with Lu. whitmani saliva and the outcome of this response after L. braziliensis infection. BALB/c mice immunized with Lu. whitmani saliva developed robust humoral and cellular immune responses, the latter characterized by an intense cellular infiltrate and production of IFN-γ and IL-10, by both CD4+ and CD8+ cells. Mice immunized as above and challenged with L. braziliensis plus Lu. whitmani saliva displayed significantly smaller lesions and parasite load at the challenge site. This protection was associated with a higher (p<0.05) IFN-γ production in response to SLA stimulation. Long-term persisting immunity was also detected in mice immunized with Lu. whitmani saliva. Furthermore, individuals residing in an endemic area for cutaneous leishmaniasis (CL) presented antibody responses to Lu. whitmani saliva. However CL patients, with active lesions, displayed a lower humoral response to Lu. whitmani saliva compared to individuals with subclinical Leishmania infection. Pre-exposure to Lu. whitmani saliva induces protection against L. braziliensis in a murine model. We also show that Lu. whitmani salivary proteins are immunogenic in naturally exposed individuals. Our results reinforce the importance of investigating the immunomodulatory effect of saliva from different species of closely related sand flies.

  2. Streptococcus Sanguis Biofilm Architecture and Its Influence on Titanium Corrosion in Enriched Artificial Saliva

    National Research Council Canada - National Science Library

    Lei Li; Shunling Li; Qing Qu; Limei Zuo; Yue He; Baolin Zhu; Cong Li

    2017-01-01

    .... We investigated the interaction between the oral Streptococcus sanguis biofilm architecture and its influence on titanium corrosion in enriched artificial saliva using electrochemical methods and microscopic study...

  3. Postnatal Identification of Zika Virus Peptides from Saliva.

    Science.gov (United States)

    Zuanazzi, D; Arts, E J; Jorge, P K; Mulyar, Y; Gibson, R; Xiao, Y; Bringel Dos Santos, M; Machado, Maria Aparecida A M; Siqueira, W L

    2017-09-01

    We explored the potential to diagnose Zika virus (ZIKV) infection by analyzing peptides in saliva during a convalescent phase of infection, long after resolution of acute disease. A 25-y-old woman clinically diagnosed with Zika fever in the first trimester was enrolled with her dizygotic twins for a 3-mo postnatal sample of saliva (9-mo after maternal infection). The female baby (A) had microcephaly while the male baby (B) was born healthy. Peptidomic analysis was completed by mass spectrometry (MS/MS), and ZIKV peptides were identified using the National Institutes of Health Zika Virus Resource database, then aligned and mapped to the ZIKV polyprotein to determine proteome coverage and phylogenetic studies. A total of 423 (mother), 607 (baby A), and 183 (baby B) unique ZIKV peptides were identified in saliva by MS/MS, providing a coverage of 67%, 84%, and 45%, respectively, of the entire ZIKV polyprotein (>3,400 amino acids). All peptides were aligned to other flaviviruses that are circulating in Brazil (dengue and yellow fever) to discard false-positive matches. Nine peptides identified were highly conserved to dengue virus. Alignment of a contiguous peptide sequence for mother/babies with the 74 ZIKV sequences suggested that the virus may have entered the oral cavity through the salivary glands, leading to an infection that persists into the postnatal period (vertical transmission). Furthermore, we identified 9 sequence variations that were unique to the baby with microcephaly (not found in the mother or the twin). This sequence information could provide a template for future neuropathogenic studies. A much larger sample size is required to determine whether sequence variation in the envelope protein significantly associates with microcephaly. Finally, from a public health perspective, it will be important to determine whether viral replication is still taking place after birth and whether the virus can be transmitted through salivary contact.

  4. Rapid Detection of the Varicella Zoster Virus in Saliva

    Science.gov (United States)

    Pierson, Duane L.; Mehta, Satish K.; Cohrs, Randall J.; Gilden, Don H.; Harding, Robert E.

    2011-01-01

    Varicella zoster virus (VZV) causes chicken pox on first exposure (usually in children), and reactivates from latency causing shingles (usually in adults). Shingles can be extremely painful, causing nerve damage, organ damage, and blindness in some cases. The virus can be life-threatening in immune-compromised individuals. The virus is very difficult to culture for diagnosis, requiring a week or longer. This invention is a rapid test for VZV from a saliva sample and can be performed in a doctor s office. The kit is small, compact, and lightweight. Detec tion is sensitive, specific, and noninvasive (no needles); only a saliva sample is required. The test provides results in minutes. The entire test is performed in a closed system, with no exposure to infectious materials. The components are made mostly of inexpensive plastic injection molded parts, many of which can be purchased off the shelf and merely assembled. All biological waste is contained for fast, efficient disposal. This innovation was made possible because of discovery of a NASA scientists flight experiment showing the presence of VZV in saliva during high stress periods and disease. This finding enables clinicians to quickly screen patients for VZV and treat the ones that show positive results with antiviral medicines. This promotes a rapid recovery, easing of pain and symptoms, and reduces chances of complications from zoster. Screening of high-risk patients could be incorporated as part of a regular physical exam. These patients include the elderly, pregnant women, and immune-compromised individuals. In these patients, VZV can be a life-threatening disease. In both high- and low-risk patients, early detection and treatment with antiviral drugs can dramatically decrease or even eliminate the clinical manifestation of disease.

  5. Kinematic viscosity of unstimulated whole saliva in healthy young adults.

    Science.gov (United States)

    Foglio-Bonda, A; Pattarino, F; Foglio-Bonda, P L

    2014-10-01

    To analyze kinematic viscosity and pH of unstimulated whole saliva, evaluate possible variations after sampling, identify any gender differences and detect possible correlations between them. The sample consisted of sixty-four healthy young adults (37 females and 27 males, mean age 25.2 years). Saliva was collected using the spitting method at 11:00 am. Kinematic viscosity was determined with a capillary viscometer (ViscoClock, Schott-Geräte Mainz, Germany) equipped with a micro-Ubbelohde capillary. Viscosity and pH were measured at a temperature of 36 °C in a thermostatic bath. Viscosity and pH data were evaluated almost simultaneously at six different times after sampling in order to identify any variations due to aging. The data were statistically analyzed using Student's t test and Wilcoxon-Mann-Whitney test. In total sample kinematic viscosity was 1.40 cSt (SD = 0.39; RSD % = 27.81), in the male and female groups was 1.33 cSt (SD = 0.35, RSD% = 26.31) and 1.45 cSt (SD = 0.41, RSD % = 28.45) respectively; the difference was not statistically significant. Viscosity decreased exponentially as a function of time after sampling then reaching a plateau around 1.12 cSt, while the pH values increased linearly. There was a trend of pH to decrease while viscosity decreases. Kinematic viscometry could be a valid tool to evaluate salivary viscosity. Degradation of saliva after sampling affects viscosity and slightly pH. The use of capillary viscometer to evaluate salivary aging needs more improvements. Further studies are required to investigate and explain the effects of different techniques to reduce the film forming on the air/liquid interface during measurement.

  6. [HBsAG in feces, urine and saliva].

    Science.gov (United States)

    Lento, F G; Tandurella, S

    1979-01-01

    After some observations about the tests of the research exposed in the literature, authors illustrate the tests for 142 patients divided into 5 groups: a) patients affected with acute viral hepatitis; b) patients affected with praegressa acute viral hepatitis; c) relatives of patients with acute viral hepatitis; d) volunteers; e) patients affected with chronic uraemia under dialisis periodic treatment. After the testing control, authors, conclude with an hipotesis: a possible epidemic function of faeces, urine saliva, in the passage of the acute viral hepatitis.

  7. Saliva-Based Biosensors: Noninvasive Monitoring Tool for Clinical Diagnostics

    Directory of Open Access Journals (Sweden)

    Radha S. P. Malon

    2014-01-01

    Full Text Available Saliva is increasingly recognised as an attractive diagnostic fluid. The presence of various disease signalling salivary biomarkers that accurately reflect normal and disease states in humans and the sampling benefits compared to blood sampling are some of the reasons for this recognition. This explains the burgeoning research field in assay developments and technological advancements for the detection of various salivary biomarkers to improve clinical diagnosis, management, and treatment. This paper reviews the significance of salivary biomarkers for clinical diagnosis and therapeutic applications, with focus on the technologies and biosensing platforms that have been reported for screening these biomarkers.

  8. Saliva-based biosensors: noninvasive monitoring tool for clinical diagnostics.

    Science.gov (United States)

    Malon, Radha S P; Sadir, Sahba; Balakrishnan, Malarvili; Córcoles, Emma P

    2014-01-01

    Saliva is increasingly recognised as an attractive diagnostic fluid. The presence of various disease signalling salivary biomarkers that accurately reflect normal and disease states in humans and the sampling benefits compared to blood sampling are some of the reasons for this recognition. This explains the burgeoning research field in assay developments and technological advancements for the detection of various salivary biomarkers to improve clinical diagnosis, management, and treatment. This paper reviews the significance of salivary biomarkers for clinical diagnosis and therapeutic applications, with focus on the technologies and biosensing platforms that have been reported for screening these biomarkers.

  9. Delivering supplemental oxygen during sedation via a saliva ejector.

    Science.gov (United States)

    Milnes, Alan R

    2002-01-01

    Intraoperative oxygen supplementation to sedated children has been shown to prevent hemoglobin desaturations even in the presence of apnea during pediatric conscious sedation. Although many practitioners deliver supplemental oxygen via a nasal hood, this method is impractical and often unsuccessful if the child is a mouth breather, has moderate adenotonsillar hypertrophy or occasionally cries during treatment (at which time there will be mouth breathing). This paper describes a method in which the saliva ejector is used to deliver supplemental oxygen to sedated children while they are receiving dental treatment. The advantages of this method and suggestions for its successful application are also included.

  10. 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

    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......) with and without fluoride in irradiated head and neck cancer patients. MATERIALS AND METHOD: Nineteen irradiated patients (median age 57 years) sucked Xerodent tablets with and without fluoride. Saliva collections were divided into three 10-min sessions in the sequence: unstimulated whole saliva, Xerodent...... of HAp crystals. RESULTS: Saliva flow rates increased significantly (15-fold) when sucking both tablets (ptablets. This was most...

  11. Rapid microfluidic solid-phase extraction system for hyper-methylated DNA enrichment and epigenetic analysis

    NARCIS (Netherlands)

    De, Arpita; Sparreboom, Wouter; van den Berg, Albert; Carlen, Edwin

    Genetic sequence and hyper-methylation profile information from the promoter regions of tumor suppressor genes are important for cancer disease investigation. Since hyper-methylated DNA (hm-DNA) is typically present in ultra-low concentrations in biological samples, such as stool, urine, and saliva,

  12. Effect of Human Saliva on Glucose Uptake by Streptococcus mutans and Other Oral Microorganisms

    Science.gov (United States)

    Germaine, Greg R.; Tellefson, Lois M.

    1981-01-01

    We examined the effects of human whole salivary supernatant and parotid fluid on glucose uptake by Streptococcus mutans, Streptococcus sanguis, Streptococcus mitis, Actinomyces viscosus, Staphylococcus aureus, and Escherichia coli. The following three effects of saliva were observed: (i) inhibition of glucose uptake (S. mutans, S. sanguis), (ii) promotion of a transient, rapid (0 to 30 s) burst of glucose uptake (S. mutans, S. sanguis), and (iii) enhancement of glucose uptake (S. mitis, A. viscosus, S. aureus, E. coli). We observed no differences between the effects of whole salivary supernatant and the effects of parotid fluid. Heat treatment (80°C, 10 min) of saliva or the addition of dithiothreitol abolished inhibition of glucose uptake. Supplementation of saliva with H2O2 potentiated inhibition of glucose uptake. S. mitis and A. viscosus, which were stimulated by saliva alone, were inhibited by H2O2-supplemented saliva; 50% inhibition of glucose uptake by S. mutans and S. mitis required ca. 10 μM H2O2 in 50% (vol/vol) saliva. Loss of the inhibitory action of saliva occurred at about 5% (vol/vol) saliva. Supplementation of saliva dilutions with SCN− and H2O2 extended the inhibitory activity to solutions containing ca. 0.2% (vol/vol) saliva. We suggest that the salivary lactoperoxidase-SCN−-H2O2 system is responsible for the inhibitory activity of saliva reported here. Furthermore, we concluded that lactoperoxidase and SCN− are present in saliva specimens in concentrations that exceed minimal inhibitory levels by factors of ca. 500 and 10 to 20, respectively. The resistance of A. viscosus, S. aureus, and E. coli to the inhibitory potential of saliva alone was probably due to the production of catalase by these organisms. The resistance of S. mitis may have been due to special effects of saliva on H2O2 accumulation by this organism compared with S. mutans and S. sanguis. The basis of saliva-dependent enhancement of glucose uptake and the basis of promotion

  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. Ancient DNA

    DEFF Research Database (Denmark)

    Willerslev, Eske; Cooper, Alan

    2004-01-01

    ancient DNA, palaeontology, palaeoecology, archaeology, population genetics, DNA damage and repair......ancient DNA, palaeontology, palaeoecology, archaeology, population genetics, DNA damage and repair...

  15. Stress corrosion cracking of NiTi in artificial saliva.

    Science.gov (United States)

    Wang, Jianqiu; Li, Nianxing; Rao, Guangbin; Han, En-Hou; Ke, Wei

    2007-02-01

    This paper aimed to study the mechanism of the cracking of orthodontic NiTi wire. Two orthodontic NiTi wires were subjected: (1) optical and scanning electron microscopy (SEM) to observe the fracture surface; (2) energy dispersive X-ray spectroscopy to determine the composition of the surface product; (3) anodic polarization to remove the surface product. Samples of NiTi alloy were subjected to the constant loading test to study the stress corrosion cracking (SCC) behavior of NiTi shape memory alloy in artificial saliva. The results showed that there were three typical areas at the fracture surface of NiTi orthodontic wire. Area '1' was a tool-made notch. Crack initiated from the root of this notch and propagated to form Area '2', which was perpendicular to the wire axis and covered by surface film. This film consisted of Na, K, Cl, P, S and O except Ni and Ti. The cracking process of NiTi alloy under the constant loading test depended on the pH of saliva and applied stress. The crack length was about 262microm, the longest at 300MPa and pH 3.0. A tool-made notch in orthodontic NiTi wires can cause SCC. At high stress and low pH, this NiTi alloy was most sensitive to cracking.

  16. Arachnids of medical importance in Brazil: main active compounds present in scorpion and spider venoms and tick saliva.

    Science.gov (United States)

    Cordeiro, Francielle A; Amorim, Fernanda G; Anjolette, Fernando A P; Arantes, Eliane C

    2015-01-01

    Arachnida is the largest class among the arthropods, constituting over 60,000 described species (spiders, mites, ticks, scorpions, palpigrades, pseudoscorpions, solpugids and harvestmen). Many accidents are caused by arachnids, especially spiders and scorpions, while some diseases can be transmitted by mites and ticks. These animals are widely dispersed in urban centers due to the large availability of shelter and food, increasing the incidence of accidents. Several protein and non-protein compounds present in the venom and saliva of these animals are responsible for symptoms observed in envenoming, exhibiting neurotoxic, dermonecrotic and hemorrhagic activities. The phylogenomic analysis from the complementary DNA of single-copy nuclear protein-coding genes shows that these animals share some common protein families known as neurotoxins, defensins, hyaluronidase, antimicrobial peptides, phospholipases and proteinases. This indicates that the venoms from these animals may present components with functional and structural similarities. Therefore, we described in this review the main components present in spider and scorpion venom as well as in tick saliva, since they have similar components. These three arachnids are responsible for many accidents of medical relevance in Brazil. Additionally, this study shows potential biotechnological applications of some components with important biological activities, which may motivate the conducting of further research studies on their action mechanisms.

  17. Glycoprotein 340 and sialic acid in minor-gland and whole saliva of children, adolescents, and adults.

    Science.gov (United States)

    Sonesson, Mikael; Ericson, Dan; Kinnby, Bertil; Wickström, Claes

    2011-12-01

    Glycoprotein 340 (gp-340) is a bacterial-binding glycoprotein found in major-gland and minor-gland saliva. Sialic acid, a common terminal structure of salivary glycoproteins, interacts with microorganisms and host ligands, as well as with free radicals. This study investigated the contents of gp-340 and sialic acid in minor-gland saliva and whole saliva of children (3 yr of age), adolescents (14 yr of age), and adults (20-25 yr of age). Labial-gland saliva and buccal-gland saliva were collected on filter paper, and unstimulated whole saliva was collected by draining into a tube. The relative amount of gp-340 and sialic acid was determined by ELISA and by enzyme-linked lectin assay (ELLA), respectively. In minor-gland saliva, no statistically significant differences in gp-340 and sialic acid were seen between the age-groups. Among adults, significantly lower amounts of gp-340 and sialic acid were seen in labial saliva compared with buccal saliva. In whole saliva, the amount of gp-340 was significantly lower among adults compared with children. No differences between genders were seen. Stable content of gp-340 and sialic acid in minor-gland saliva across the age-groups, and a higher content of gp-340 in the whole saliva of the youngest age-group (3-yr-olds) compared with the adult group, may reflect that those components are vital innate factors of immunity in children's saliva. © 2011 Eur J Oral Sci.

  18. Repeated extraction of DNA from FTA cards

    DEFF Research Database (Denmark)

    Stangegaard, Michael; Ferrero, Laura; Børsting, Claus

    2011-01-01

    Extraction of DNA using magnetic bead based techniques on automated DNA extraction instruments provides a fast, reliable and reproducible method for DNA extraction from various matrices. However, the yield of extracted DNA from FTA-cards is typically low. Here, we demonstrate that it is possible...... to repeatedly extract DNA from the processed FTA-disk. The method increases the yield from the nanogram range to the microgram range....

  19. Proteomic analysis of human whole and parotid salivas following stimulation by different tastes

    NARCIS (Netherlands)

    Neyraud, E.; Sayd, T.; Morzel, M.; Dransfield, E.

    2006-01-01

    Whole and parotid salivas, collected after stimulation with tastants, were analyzed by 2D electrophoresis and mass spectrometry. In whole saliva, the number of proteins affected by taste stimulation increased in the order sweet < umami < bitter < acid. Annexin A1 and calgranulin A, involved in

  20. The effect of saliva composition on texture perception of semi-solids

    NARCIS (Netherlands)

    Engelen, L.; Keybus, van den P.A.M.; Wijk, de R.A.; Veerman, E.C.I.; Nieuw Amerongen, A.V.; Bosman, F.; Prinz, J.F.; Bilt, van der A.

    2007-01-01

    Saliva is expected to be of significance for the perception of food stimuli in the mouth. Mixing the food with saliva, including breakdown and dilution, is considered to be of large importance for semi-solids as these products are masticated without chewing. It is known that there are large

  1. Investigating the Hydrolysis of Starch Using "a"-Amylase Contained in Dishwashing Detergent and Human Saliva

    Science.gov (United States)

    Munegumi, Toratane; Inutsuka, Masato; Hayafuji, Yukitaka

    2016-01-01

    Although saliva has commonly been used to teach about digestion by organisms, the phenomenon of digestion is actually caused by enzymes as catalytic substances. This activity explores the hydrolysis of starch by "a"-amylase in cleaning materials as well as a comparison with the similar reaction using human saliva. The fact that the…

  2. Comparison of two chair-side tests for enumeration of Mutans Streptococci in saliva

    DEFF Research Database (Denmark)

    Twetman, Lisa; Twetman, Svante

    2014-01-01

    , referred to a maxillofacial hospital clinic with a caries history. Stimulated whole saliva samples were collected and the number of MS was assessed with the Dentocult-SM Strip Mutans (DSM) and the Saliva-Check Mutans (SCM). The outcome was compared with conventional anaerobic laboratory cultivation...

  3. Saliva DHEAS Changes in Patients Suffering from Psychopathological Disorders Arising from Bullying at Work

    Science.gov (United States)

    Lac, Gerard; Dutheil, Frederic; Brousse, Georges; Triboulet-Kelly, Celine; Chamoux, Alain

    2012-01-01

    Background: Psychological disorders arising from bullying at work (BW) are common. The relationship between these disorders and putative markers is not well established. Aims: To measure saliva dehydroepiandrosterone sulphate (DHEAS) and saliva cortisol as putative markers in individuals suffering from BW. Methods: Forty one subjects suffering…

  4. Effects of isoflurane anesthesia and pilocarpine on rat parotid saliva flow

    DEFF Research Database (Denmark)

    Knudsen, Jacob Dronninglund; Nauntofte, Birgitte; Josipovic, M

    2011-01-01

    rats was 50% slower than that of the sham-irradiated rats. In conclusion, 1.5% isoflurane was found to be a good compromise between proper anesthesia and isoflurane-induced inhibition of saliva secretion. Pilocarpine induces saliva secretion in a dose-dependent matter, with supra-maximal stimulation...

  5. SELDI-TOF-MS of saliva : Methodology and pre-treatment effects

    NARCIS (Netherlands)

    Schipper, R.G.; Loof, D.; Groot, de J.; Harthoorn, L.F.; Dransfield, E.; Heerde, van W.

    2007-01-01

    Interest in saliva as a diagnostic fluid for monitoring general health and for early diagnosis of disease has increased in the last few years. In particular, efforts have focused on the generation of protein maps of saliva using advanced proteomics technology. Surface-enhanced

  6. Validation and quality control of ELISAs for the use with human saliva samples.

    Science.gov (United States)

    Jaedicke, Katrin M; Taylor, John J; Preshaw, Philip M

    2012-03-30

    Enzyme-linked immunosorbent assays (ELISAs) have proven to be a powerful tool for fast and reliable sample analysis, in both clinical diagnostics and in research. Most assays are now available for use with a range of different analytical fluids, including serum, plasma or urine. In recent years, saliva has drawn attention as a potentially valuable diagnostic fluid; however few ELISAs have been validated for use with saliva, or their validation is often incomplete. Saliva has a number of different physical characteristics than, for example, cell culture medium or serum and assuming an ELISA which works well with serum samples will also do so with saliva potentially could lead to erroneous data and conclusions. In this report, we provide a detailed protocol to validate any ELISA for use with saliva samples and show the results of validation procedures for 13 ELISAs for using saliva. Our findings suggest that the majority of ELISAs work reliably with saliva, even if the assay was not specifically designed for this biological fluid. However, we also report a few cases where recovery or intra-and inter-assay variations were unexpectedly high, emphasising the importance of performing a validation procedure for each assay before using it with saliva to ensure accurate and reliable data. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. Total Protein of Whole Saliva as a Biomarker of Anaerobic Threshold

    Science.gov (United States)

    Bortolini, Miguel Junior Sordi; De Agostini, Guilherme Gularte; Reis, Ismair Teodoro; Lamounier, Romeu Paulo Martins Silva; Blumberg, Jeffrey B.; Espindola, Foued Salmen

    2009-01-01

    Saliva provides a convenient and noninvasive matrix for assessing specific physiological parameters, including some biomarkers of exercise. We investigated whether the total protein concentration of whole saliva (TPWS) would reflect the anaerobic threshold during an incremental exercise test. After a warm-up period, 13 nonsmoking men performed a…

  8. Determination of epirubicin and its metabolite epirubicinol in saliva and plasma by HPLC

    NARCIS (Netherlands)

    Dodde, WIW; Maring, JG; Hendriks, G; Wachters, FM; Groen, HJM; de Vries, EGE; Uges, DRA

    We present a high-performance liquid chromatography (HPLC) method suitable for the analysis of epirubicin and its metabolite epirubicinol in saliva and plasma. Preparation of saliva and plasma samples was performed by extraction of analytes with a chloroform: 2-propanol mixture (6:1, vol/vol) and

  9. Saliva initiates the formation of pro-inflammatory macrophages in vitro.

    Science.gov (United States)

    Pourgonabadi, Solmaz; Müller, Heinz-Dieter; Mendes, João Rui; Gruber, Reinhard

    2017-01-01

    Saliva can support oral wound healing, a process that requires a temporary inflammatory reaction. We have reported previously that saliva provokes a strong inflammatory response in oral fibroblasts. Bone marrow cells also give rise to macrophages, a heterogeneous subset of cell population involved in wound healing. Lipopolysaccharide (LPS) and interleukin 4 (IL-4) induce activation of pro-(M1), and anti-(M2) inflammatory macrophages, respectively. Yet, the impact of saliva on programming bone marrow cells into either M1 or M2 macrophages remains unclear . Herein, we examined whether sterile saliva affects the in vitro process of macrophage polarization based on murine bone marrow cultures and RAW264.7 mouse macrophages. We report that sterile saliva, similar to lipopolysaccharides, provoked a robust activation of the M1 phenotype which is characterized by a strong increase of the respective genes IL-12 and IL-6, based on a real-time gene expression analysis, and for IL-6 with immunoassay. Arginase-1 and Ym1, both genes characteristic for the M2 phenotype, were not considerably modulated by saliva. Inhibition of TLR4 signaling with TAK-242, blocking NFκB signaling with Bay 11-7085, but also autoclaving saliva greatly reduced the development of the M1 phenotype. These data suggest that saliva activates the TLR4 dependent polarization into pro-inflammatory M1 macrophages in vitro. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Comparative assessment of saliva and plasma for drug bioavailability and bioequivalence studies in humans

    Directory of Open Access Journals (Sweden)

    Nasir M. Idkaidek

    2017-07-01

    Conclusion: Our results suggest that there is a potential in BA/BE studies for saliva to be considered as a surrogate for plasma concentration, which goes along with drug regulations. The use of saliva instead of plasma in such studies makes them non-invasive, easy and with a lower clinical burden.

  11. Watery Saliva Secreted by the Grain Aphid Sitobion avenae Stimulates Aphid Resistance in Wheat.

    Science.gov (United States)

    Zhang, Yong; Fan, Jia; Francis, Frédéric; Chen, Julian

    2017-10-11

    Infestation with Sitobion avenae induces localized defense responses in wheat; in this study, the role of S. avenae watery saliva in resistance induction was examined by infiltrating aphid saliva into wheat leaves. After feeding S. avenae on an artificial diet for 48 h, we first collected watery saliva from them and then separated the salivary proteins using one-dimensional gel electrophoresis. Gene expression studies showed that infiltration of S. avenae watery saliva in wheat leaves induced a strong salicylic acid-responsive defense but moderate jasmonic acid-dependent defense. Feeding on wheat leaves infiltrated with aphid saliva, compared with untreated leaves, significantly decreased the number of nymphs produced per day and the intrinsic rate of increase of the population of S. avenae. In a choice test against untreated wheat, saliva-infiltrated wheat had repellent effects on aphids. Additionally, electrical penetration graph results showed that the feeding behavior of S. avenae on saliva-treated wheat was negatively affected compared with that on untreated wheat. These findings provided direct evidence that salivary components of S. avenae are involved in the induction of wheat resistance against aphids and further demonstrated the important roles of watery saliva in aphid-plant interactions.

  12. Relationships between nicotine and cotinine concentrations in maternal milk and saliva.

    Science.gov (United States)

    Jacob, Nelly; Golmard, Jean-Louis; Berlin, Ivan

    2015-08-01

    Breastfeeding may be impaired due to nicotine excreted into the milk of smoking mothers. We investigated the relationships between nicotine and cotinine concentrations in maternal milk and saliva among breastfeeding smokers. The 41 mothers reported their cigarette consumption between waking up and milk and saliva sampling. The median sampling time took place four days after delivery. Nicotine and cotinine concentrations were determined by liquid chromatography and UV detection, after a single-step saliva or three-step milk liquid-to-liquid extraction. The median (interquartile range) concentrations in milk and saliva were 7 (6-22) and 27 (4-207) μg/L for nicotine and 24 (5-111) and 22 (4-120) μg/L for cotinine, respectively. Milk cotinine was positively associated with saliva cotinine (p saliva nicotine concentration (p = 0.0017) and cigarette consumption (p = 0.0023, model R(2) = 0.63). Saliva nicotine concentration was not a very good estimate of milk nicotine concentration in breastfeeding mothers. Saliva cotinine concentration may be used instead of milk cotinine concentration to estimate tobacco or nicotine exposure among breastfed neonates or infants. ©2015 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.

  13. Measurement of Creatine kinase and Aspartate aminotransferase in saliva of dogs: a pilot study.

    Science.gov (United States)

    Tvarijonaviciute, Asta; Barranco, Tomas; Rubio, Monica; Carrillo, Jose Maria; Martinez-Subiela, Silvia; Tecles, Fernando; Carrillo, Juana Dolores; Cerón, José J

    2017-06-09

    Muscle enzymes in saliva have been reported to be possible markers of heart and muscle damage in humans. The aim of this study was to assess if Creatine kinase (CK) and Aspartate aminotransferase (AST) activities could be measured in canine saliva, and to evaluate their possible changes in situations of muscle damage. The spectrophotometric assays for CK and AST measurement in saliva of dogs showed intra- and inter-assay imprecision lower than 1 and 16% and coefficients of correlation close to 1 in linearity under dilution tests. Healthy dogs showed activities in saliva of CK between 27 and 121 U/L and AST between 46 and 144 U/L, whereas in saliva of dogs with muscle damage CK ranged between 132 and 3862 U/L and AST between 154 and 4340 U/L. Positive moderate correlations were found between saliva and serum activities of the two enzymes (CK, r = 0.579; P = 0.001; AST, r = 0.674; P = 0.001). CK and AST activities can be measured in canine saliva with commercially available spectrophotometric assays. In addition these enzymes show higher values in saliva of dogs with muscle damage and their values are moderately correlated with those of serum.

  14. Treatment of xerostomia with polymer-based saliva substitutes in patients with Sjogren's syndrome

    NARCIS (Netherlands)

    vanderReijden, WA; vanderKwaak, H; Vissink, A; Veerman, ECI; Amerongen, AVN

    Objective. To determine the efficacy of 3 types of polymer-based saliva substitutes in reducing oral dryness in patients with Sjogren's syndrome (SS). Methods. Subjective efficacy of 3 different saliva substitutes (determined by self-administered questionnaire) was evaluated in a double-blind,

  15. Evaluation of physio-chemical properties of saliva and comparison of its relation with dental caries.

    Science.gov (United States)

    Dogra, Subha; Bhayya, Deepak; Arora, Ruchi; Singh, Deepesh; Thakur, Dashmesh

    2013-01-01

    Objective was to evaluate the relationship between physio-chemical properties of saliva such as flow rate, buffering capacity, pH, Streptococcus mutans in saliva and its relationship with dental caries. Eighty children were evaluated for physio-chemical properties of saliva, out which 40 were caries-active (group 1) and 40 caries-free (control group). Caries status of each child was scored by using DMFS and dfs indices to get a combined DMFS and dfs score. The physio-chemical properties were evaluated using Saliva Check (GC Asia Dental Pte Ltd- India) and streptococcus mutans using Dentocult SM Strip Mutans. Flow rate, pH, and buffering capacity of saliva in caries-active children were decreased but not statistically significant. The Streptococcus mutant count of saliva was increased significantly in caries-active children. The physio-chemical properties of saliva like pH, buffering capacity, salivary flow rate, concentration of various components like proteins, calcium and antioxidant defense system play a major role in the development of caries. Hence, more clinical and laboratory studies are needed to determine the exact relationship between these physio-chemical properties of saliva and dental caries.

  16. Evaluation of a rapid test for HIV antibodies in saliva and blood ...

    African Journals Online (AJOL)

    Saliva specimens can be readily collected from any individual, and there is a reduction in hazard risk. Anti-HIV saliva testing using the test strip methodology is recommended for South Africa, particularly in high-risk situations such as the paediatric and forensic medicine settings. A larger field study obtaining specimens from ...

  17. 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

  18. Influence of mastication and saliva on aroma release in a model mouth system

    NARCIS (Netherlands)

    Ruth, van S.M.; Roozen, J.P.

    2000-01-01

    The influence of mastication, saliva composition and saliva volume on aroma release from rehydrated diced bell peppers and French beans was studied in a model mouth system. Released volatile compounds were analysed by gas chromatography combined with sniffing port and flame ionisation detection.

  19. Dried saliva spots : A robust method for detecting Streptococcus pneumoniae carriage by PCR

    NARCIS (Netherlands)

    Krone, Cassandra L.; Oja, Anna E.; van de Groep, Kirsten|info:eu-repo/dai/nl/413649776; Sanders, Elisabeth A M|info:eu-repo/dai/nl/126771960; Bogaert, Debby|info:eu-repo/dai/nl/264105834; Trzcinski, Krzysztof|info:eu-repo/dai/nl/323349609

    2016-01-01

    The earliest studies in the late 19th century on Streptococcus pneumoniae (S. pneumoniae) carriage used saliva as the primary specimen. However, interest in saliva declined after the sensitive mouse inoculation method was replaced by conventional culture, which made isolation of pneumococci from the

  20. Detection of Helicobacter pylori urease antigen in saliva in patients with different gastric H. pylori status

    Directory of Open Access Journals (Sweden)

    Mounia El Khadir

    2016-07-01

    Conclusion: This study demonstrated a low detection rate of H. pylori antigens in saliva compared with the presence of this bacterium in gastric mucosa, suggesting that saliva cannot be used as a suitable sample for the diagnosis of H. pylori in our study population.

  1. Handheld Device Adapted to Smartphone Cameras for the Measurement of Sodium Ion Concentrations at Saliva-Relevant Levels via Fluorescence

    Directory of Open Access Journals (Sweden)

    Michelle Lipowicz

    2015-06-01

    Full Text Available The use of saliva sampling as a minimally-invasive means for drug testing and monitoring physiology is a subject of great interest to researchers and clinicians. This study describes a new optical method based on non-axially symmetric focusing of light using an oblate spheroid sample chamber. The device is simple, lightweight, low cost and is easily attached to several different brands/models of smartphones (Apple, Samsung, HTC and Nokia for the measurement of sodium ion levels at physiologically-relevant saliva concentrations. The sample and fluorescent reagent solutions are placed in a specially-designed, lightweight device that excludes ambient light and concentrates 470-nm excitation light, from a low-power photodiode, within the sample through non-axially-symmetric refraction. The study found that smartphone cameras and post-image processing quantitated sodium ion concentration in water over the range of 0.5–10 mM, yielding best-fit regressions of the data that agree well with a data regression of microplate luminometer results. The data suggest that fluorescence can be used for the measurement of salivary sodium ion concentrations in low-resource or point-of-care settings. With further fluorescent assay testing, the device may find application in a variety of enzymatic or chemical assays.

  2. Handheld Device Adapted to Smartphone Cameras for the Measurement of Sodium Ion Concentrations at Saliva-Relevant Levels via Fluorescence.

    Science.gov (United States)

    Lipowicz, Michelle; Garcia, Antonio

    2015-06-02

    The use of saliva sampling as a minimally-invasive means for drug testing and monitoring physiology is a subject of great interest to researchers and clinicians. This study describes a new optical method based on non-axially symmetric focusing of light using an oblate spheroid sample chamber. The device is simple, lightweight, low cost and is easily attached to several different brands/models of smartphones (Apple, Samsung, HTC and Nokia) for the measurement of sodium ion levels at physiologically-relevant saliva concentrations. The sample and fluorescent reagent solutions are placed in a specially-designed, lightweight device that excludes ambient light and concentrates 470-nm excitation light, from a low-power photodiode, within the sample through non-axially-symmetric refraction. The study found that smartphone cameras and post-image processing quantitated sodium ion concentration in water over the range of 0.5-10 mM, yielding best-fit regressions of the data that agree well with a data regression of microplate luminometer results. The data suggest that fluorescence can be used for the measurement of salivary sodium ion concentrations in low-resource or point-of-care settings. With further fluorescent assay testing, the device may find application in a variety of enzymatic or chemical assays.

  3. Metaproteomics of saliva identifies human protein markers specific for individuals with periodontitis and dental caries compared to orally healthy controls

    DEFF Research Database (Denmark)

    Belstrøm, Daniel; Jersie-Christensen, Rosa R; Lyon, David

    2016-01-01

    BACKGROUND: The composition of the salivary microbiota has been reported to differentiate between patients with periodontitis, dental caries and orally healthy individuals. To identify characteristics of diseased and healthy saliva we thus wanted to compare saliva metaproteomes from patients with...

  4. Rapid detection of lactate dehydrogenase and genotyping of Plasmodium falciparum in saliva of children with acute uncomplicated malaria.

    Science.gov (United States)

    Gbotosho, Grace O; Happi, Christian T; Folarin, Onikepe; Keyamo, Ochuko; Sowunmi, Akintunde; Oduola, Ayoade M J

    2010-09-01

    The diagnosis of malaria in biological fluids other than blood using non-invasive, rapid diagnostic techniques provides a valuable approach in case management and epidemiological studies of malaria. Rapid detection of Plasmodium falciparum lactate dehydrogenase (pLDH) in saliva samples from 130 of 144 children with microscopically confirmed P. falciparum infection was evaluated using Optimal-IT dipsticks. Genotyping of parasites was also performed in saliva and blood samples from a cohort of patients by polymerase chain reaction (PCR). The sensitivity of the dipstick in whole-blood, whole-saliva, or supernatant of spun saliva samples was 97.2%, 77.9%, and 48.4%, respectively. The sensitivity of the dipstick in whole-saliva samples was significantly higher than in supernatant of spun saliva samples (P saliva samples, respectively. This finding shows rapid detection of pLDH in patient saliva.

  5. Detection of Helicobacter spp. in the saliva of dogs with gastritis.

    Science.gov (United States)

    Jankowski, M; Spużak, J; Kubiak, K; Glińska-Suchocka, K; Biernat, M

    2016-01-01

    The aim of this study was to identify the species and determine the prevalence of gastric Helicobacter in the saliva of dogs with gastritis. The study was carried out on 30 dogs of different breeds, genders and ages, which were diagnosed with gastritis. The nested-PCR method was used to detect Helicobacter spp. in saliva. Helicobacter bacteria were found in the saliva samples of 23 (76.6%) dogs. Helicobacter heilmannii was the most commonly detected species of gastric Helicobacter spp. in canine saliva, and was found in 22 (73.3%) cases. The results indicate that gastric Helicobacter spp. occurs relatively frequently in dogs with gastritis. Moreover, the saliva of dogs with gastritis may be a source of Helicobacter spp. infection for humans and other animals. However, further studies are needed to confirm this finding as the PCR method does not distinguish active from inactive infections.

  6. Microbial profile comparisons of saliva, pooled and site-specific subgingival samples in periodontitis patients

    DEFF Research Database (Denmark)

    Belstrøm, Daniel; Sembler-Møller, Maria Lynn; Grande, Maria Anastasia

    2017-01-01

    OBJECTIVES: The purpose of this study was to compare microbial profiles of saliva, pooled and site-specific subgingival samples in patients with periodontitis. We tested the hypotheses that saliva can be an alternative to pooled subgingival samples, when screening for presence of periopathogens....... DESIGN: Site specific subgingival plaque samples (n = 54), pooled subgingival plaque samples (n = 18) and stimulated saliva samples (n = 18) were collected from 18 patients with generalized chronic periodontitis. Subgingival and salivary microbiotas were characterized by means of HOMINGS (Human Oral...... to an AUC of 0.76 (sensitivity: 0.56, specificity: 0.94) in pooled subgingival samples. CONCLUSIONS: Site-specific presence of periodontal pathogens was detected with comparable accuracy in stimulated saliva samples and pooled subgingival plaque samples. Consequently, saliva may be a reasonable surrogate...

  7. Ultra-deep and quantitative saliva proteome reveals dynamics of the oral microbiome

    DEFF Research Database (Denmark)

    Grassl, Niklas; Kulak, Nils Alexander; Pichler, Garwin

    2016-01-01

    , disruptions in saliva secretion and changes in the oral microbiome contribute to conditions such as tooth decay and respiratory tract infections. Here we set out to quantitatively map the saliva proteome in great depth with a rapid and in-depth mass spectrometry-based proteomics workflow. METHODS: We used...... recent improvements in mass spectrometry (MS)-based proteomics to develop a rapid workflow for mapping the saliva proteome quantitatively and at great depth. Standard clinical cotton swabs were used to collect saliva form eight healthy individuals at two different time points, allowing us to study inter......-individual differences and interday changes of the saliva proteome. To accurately identify microbial proteins, we developed a method called "split by taxonomy id" that prevents peptides shared by humans and bacteria or between different bacterial phyla to contribute to protein identification. RESULTS: Microgram protein...

  8. Short communication: HIV type 1 escapes inactivation by saliva via rapid escape into oral epithelial cells.

    Science.gov (United States)

    Dietrich, Elizabeth A; Gebhard, Kristin H; Fasching, Claudine E; Giacaman, Rodrigo A; Kappes, John C; Ross, Karen F; Herzberg, Mark C

    2012-12-01

    Saliva contains anti-HIV-1 factors, which show unclear efficacy in thwarting mucosal infection. When incubated in fresh, unfractionated whole saliva, infectious HIV-1 IIIb and BaL (X4- and R5-tropic, respectively) persisted from 4 to at least 30 min in a saliva concentration-dependent manner. In salivary supernatant for up to 6 h, both infectious HIV-1 strains "escaped" into immortalized oral epithelial cells; infectious BaL showed selectively enhanced escape in the presence of saliva. Fluorescently labeled HIV-1 virus-like particles entered oral epithelial cells within minutes of exposure. Using a previously unrecognized mechanism, therefore, strains of HIV-1 escape inactivation by saliva via rapid uptake into oral epithelial cells.

  9. Oral Administration of Lactobacillus plantarum 299v Reduces Cortisol Levels in Human Saliva during Examination Induced Stress: A Randomized, Double-Blind Controlled Trial

    Directory of Open Access Journals (Sweden)

    Hannah Andersson

    2016-01-01

    Full Text Available Objective. To clarify the effect of Lactobacillus plantarum 299v on the salivary cortisol and salivary IgA levels in young adults under examination stress. Design. Forty-one students with an upcoming academic exam were included in a randomized double-blind, placebo-controlled study. The probiotic bacteria or the placebo product was administered in capsules once a day during 14 days. Saliva was collected and a perceived stress test was filled out at each sampling occasion. Saliva was collected for cortisol analysis by Electrochemiluminescence Immunoassay (ECLI and salivary IgA was analysed by Enzyme-Linked Immunosorbent Assay (ELISA. Abundance of lactobacilli was evaluated by cultivation of saliva on selective medium and identification of L. plantarum 299v was done on randomly selected colonies by a random amplification of polymorphic DNA (RAPD typing. Results. A significant difference in cortisol levels was found between the treatment group and the placebo group (P<0.05, together with a significant increase in levels of lactobacilli in the treatment group compared with the placebo group (P<0.001. No significant changes were found for salivary IgA. Conclusion. A probiotic bacterium with ability to reduce symptoms of irritable bowel syndrome (IBS prohibited increased levels of the stress marker cortisol during the examination period. The registration number of the study is NCT02974894, and the study is registered at ClinicalTrials.gov.

  10. Oral Administration of Lactobacillus plantarum 299v Reduces Cortisol Levels in Human Saliva during Examination Induced Stress: A Randomized, Double-Blind Controlled Trial.

    Science.gov (United States)

    Andersson, Hannah; Tullberg, Cecilia; Ahrné, Siv; Hamberg, Kristina; Lazou Ahrén, Irini; Molin, Göran; Sonesson, Mikael; Håkansson, Åsa

    2016-01-01

    Objective. To clarify the effect of Lactobacillus plantarum 299v on the salivary cortisol and salivary IgA levels in young adults under examination stress. Design. Forty-one students with an upcoming academic exam were included in a randomized double-blind, placebo-controlled study. The probiotic bacteria or the placebo product was administered in capsules once a day during 14 days. Saliva was collected and a perceived stress test was filled out at each sampling occasion. Saliva was collected for cortisol analysis by Electrochemiluminescence Immunoassay (ECLI) and salivary IgA was analysed by Enzyme-Linked Immunosorbent Assay (ELISA). Abundance of lactobacilli was evaluated by cultivation of saliva on selective medium and identification of L. plantarum 299v was done on randomly selected colonies by a random amplification of polymorphic DNA (RAPD) typing. Results. A significant difference in cortisol levels was found between the treatment group and the placebo group (P stress marker cortisol during the examination period. The registration number of the study is NCT02974894, and the study is registered at ClinicalTrials.gov.

  11. [Verification and Validation on Single Nucleotide Polymorphism Analysis of Alcohol Metabolism-Related Genes ADH1B and ALDH2, Using Dried-Saliva Samples].

    Science.gov (United States)

    Murata, Shigenori; Hayashida, Mariko; Ishiguro-Tanaka, Yuko; Imazeki, Hiromi; Hayashi, Emiko; Yokoyama, Akira; Kinoshita, Kenji

    2015-11-01

    We have developed a new method for unprocessed biological specimens as templates directly into the TaqMan assay. Saliva was needed to be put on a water-soluble paper and dried, because foreign substances, such as a filter paper, hinder fluorescence detection through the assay. Genotyping of alcohol metabolism-related genes ADH1B (rs1229984) and ALDH2 (rs671) polymorphisms was, subsequently, performed by TaqMan PCR assay using dried saliva in the present investigation. The optimized technique was tested on 114 samples of alcoholic patients. The PCR-RFLP methods with purified DNA from blood samples were employed for validation of the assay. Upon validation, complete concordance was observed between the two independent results. These results highlight the ability of TaqMan PCR assays using dried saliva on water-soluble paper in genotyping of ADH1B and ALDH2 genes. Our results showed a rapid, simple, reliable, and cost-effective method for SNP genotyping of mutations in ADH1B and ALDH2 genes. This will be very useful for large-scale association studies in various fields. [Original].

  12. Endocannabinoids measurement in human saliva as potential biomarker of obesity.

    Directory of Open Access Journals (Sweden)

    Isabelle Matias

    Full Text Available BACKGROUND: The discovery of the endocannabinoid system and of its role in the regulation of energy balance has significantly advanced our understanding of the physiopathological mechanisms leading to obesity and type 2 diabetes. New knowledge on the role of this system in humans has been acquired by measuring blood endocannabinoids. Here we explored endocannabinoids and related N-acylethanolamines in saliva and verified their changes in relation to body weight status and in response to a meal or to body weight loss. METHODOLOGY/PRINCIPAL FINDINGS: Fasting plasma and salivary endocannabinoids and N-acylethanolamines were measured through liquid mass spectrometry in 12 normal weight and 12 obese, insulin-resistant subjects. Salivary endocannabinoids and N-acylethanolamines were evaluated in the same cohort before and after the consumption of a meal. Changes in salivary endocannabinoids and N-acylethanolamines after body weight loss were investigated in a second group of 12 obese subjects following a 12-weeks lifestyle intervention program. The levels of mRNAs coding for enzymes regulating the metabolism of endocannabinoids, N-acylethanolamines and of cannabinoid type 1 (CB(1 receptor, alongside endocannabinoids and N-acylethanolamines content, were assessed in human salivary glands. The endocannabinoids 2-arachidonoylglycerol (2-AG, N-arachidonoylethanolamide (anandamide, AEA, and the N-acylethanolamines (oleoylethanolamide, OEA and palmitoylethanolamide, PEA were quantifiable in saliva and their levels were significantly higher in obese than in normal weight subjects. Fasting salivary AEA and OEA directly correlated with BMI, waist circumference and fasting insulin. Salivary endocannabinoids and N-acylethanolamines did not change in response to a meal. CB(1 receptors, ligands and enzymes were expressed in the salivary glands. Finally, a body weight loss of 5.3% obtained after a 12-weeks lifestyle program significantly decreased salivary AEA

  13. Quantitative detection of PfHRP2 in saliva of malaria patients in the Philippines.

    Science.gov (United States)

    Fung, Andrew O; Damoiseaux, Robert; Grundeen, Sarah; Panes, Jonnas L; Horton, Daniel H; Judy, Jack W; Moore, Theodore B

    2012-05-25

    Malaria is a global health priority with a heavy burden of fatality and morbidity. Improvements in field diagnostics are needed to support the agenda for malaria elimination. Saliva has shown significant potential for use in non-invasive diagnostics, but the development of off-the-shelf saliva diagnostic kits requires best practices for sample preparation and quantitative insight on the availability of biomarkers and the dynamics of immunoassay in saliva. This pilot study measured the levels of the PfHRP2 in patient saliva to inform the development of salivary diagnostic tests for malaria. Matched samples of blood and saliva were collected between January and May, 2011 from eight patients at Palawan Baptist Hospital in Roxas, Palawan, Philippines. Parasite density was determined from thick-film blood smears. Concentrations of PfHRP2 in saliva of malaria-positive patients were measured using a custom chemiluminescent ELISA in microtitre plates. Sixteen negative-control patients were enrolled at UCLA. A substantive difference between this protocol and previous related studies was that saliva samples were stabilized with protease inhibitors. Of the eight patients with microscopically confirmed P. falciparum malaria, seven tested positive for PfHRP2 in the blood using rapid diagnostic test kits, and all tested positive for PfHRP2 in saliva. All negative-control samples tested negative for salivary PfHRP2. On a binary-decision basis, the ELISA agreed with microscopy with 100 % sensitivity and 100 % specificity. Salivary levels of PfHRP2 ranged from 17 to 1,167 pg/mL in the malaria-positive group. Saliva is a promising diagnostic fluid for malaria when protein degradation and matrix effects are mitigated. Systematic quantitation of other malaria biomarkers in saliva would identify those with the best clinical relevance and suitability for off-the-shelf diagnostic kits.

  14. Quantitative detection of PfHRP2 in saliva of malaria patients in the Philippines

    Directory of Open Access Journals (Sweden)

    Fung Andrew O

    2012-05-01

    Full Text Available Abstract Background Malaria is a global health priority with a heavy burden of fatality and morbidity. Improvements in field diagnostics are needed to support the agenda for malaria elimination. Saliva has shown significant potential for use in non-invasive diagnostics, but the development of off-the-shelf saliva diagnostic kits requires best practices for sample preparation and quantitative insight on the availability of biomarkers and the dynamics of immunoassay in saliva. This pilot study measured the levels of the PfHRP2 in patient saliva to inform the development of salivary diagnostic tests for malaria. Methods Matched samples of blood and saliva were collected between January and May, 2011 from eight patients at Palawan Baptist Hospital in Roxas, Palawan, Philippines. Parasite density was determined from thick-film blood smears. Concentrations of PfHRP2 in saliva of malaria-positive patients were measured using a custom chemiluminescent ELISA in microtitre plates. Sixteen negative-control patients were enrolled at UCLA. A substantive difference between this protocol and previous related studies was that saliva samples were stabilized with protease inhibitors. Results Of the eight patients with microscopically confirmed P. falciparum malaria, seven tested positive for PfHRP2 in the blood using rapid diagnostic test kits, and all tested positive for PfHRP2 in saliva. All negative-control samples tested negative for salivary PfHRP2. On a binary-decision basis, the ELISA agreed with microscopy with 100 % sensitivity and 100 % specificity. Salivary levels of PfHRP2 ranged from 17 to 1,167 pg/mL in the malaria-positive group. Conclusion Saliva is a promising diagnostic fluid for malaria when protein degradation and matrix effects are mitigated. Systematic quantitation of other malaria biomarkers in saliva would identify those with the best clinical relevance and suitability for off-the-shelf diagnostic kits.

  15. Saliva from Obese Individuals Suppresses the Release of Aroma Compounds from Wine

    Science.gov (United States)

    Piombino, Paola; Genovese, Alessandro; Esposito, Silvia; Moio, Luigi; Cutolo, Pier Paolo; Chambery, Angela; Severino, Valeria; Moneta, Elisabetta; Smith, Daniel P.; Owens, Sarah M.; Gilbert, Jack A.; Ercolini, Danilo

    2014-01-01

    Background Recent evidence suggests that a lower extent of the retronasal aroma release correspond to a higher amount of ad libitum food intake. This has been regarded as one of the bases of behavioral choices towards food consumption in obese people. In this pilot study we investigated the hypothesis that saliva from obese individuals could be responsible for an alteration of the retro-nasal aroma release. We tested this hypothesis in vitro, by comparing the release of volatiles from a liquid food matrix (wine) after its interaction with saliva from 28 obese (O) and 28 normal-weight (N) individuals. Methods and Findings Amplicon sequencing of the 16S rRNA V4 region indicated that Firmicutes and Actinobacteria were more abundant in O, while Proteobacteria and Fusobacteria dominated in N. Streptococcaceae were significantly more abundant in the O subjects and constituted 34% and 19% on average of the saliva microbiota of O and N subjects, respectively. The Total Antioxidant Capacity was higher in O vs N saliva samples. A model mouth system was used to test whether the in-mouth wine aroma release differs after the interaction with O or N saliva. In O samples, a 18% to 60% significant decrease in the mean concentration of wine volatiles was detected as a result of interaction with saliva, compared with N. This suppression was linked to biochemical differences in O and N saliva composition, which include protein content. Conclusion Microbiological and biochemical differences were found in O vs N saliva samples. An impaired retronasal aroma release from white wine was detected in vitro and linked to compositional differences between saliva from obese and normal-weight subjects. Additional in vivo investigations on diverse food matrices could contribute to understanding whether a lower olfactory stimulation due to saliva composition can be a co-factor in the development/maintenance of obesity. PMID:24465618

  16. Assessing cortisol and dehydroepiandrosterone (DHEA) in saliva: effects of collection method.

    Science.gov (United States)

    Gallagher, Peter; Leitch, Melville M; Massey, Anna E; McAllister-Williams, R Hamish; Young, Allan H

    2006-09-01

    An increasing number of studies are utilizing saliva sampling as a method of assessing adrenal steroid secretion. Saliva samples have certain advantages over plasma, being non-invasive and easily collected. However, some methods of collection may compromise the accuracy of the assay, particularly those which employ aids to stimulate saliva production. We sought to compare the accuracy of cortisol and dehydroepiandrosterone (DHEA) measurement by examining the association between plasma levels, saliva and saliva collected using a citric acid-treated salivette device. Twenty six healthy male volunteers were recruited for the study. To increase the range of steroid levels in the samples collected, half the subjects were pre-treated with hydrocortisone (20mg, twice a day for 7 days) and half with placebo. Saliva samples were then collected from each subject using both a 'passive drool' method and a citric acid-treated salivette. A plasma sample was also collected. Cortisol and DHEA levels were measured by radioimmunoassay. For cortisol levels, both methods of saliva collection correlated highly with plasma levels and with each other (r 0.85; R(2) 0.72 for all). For DHEA levels, only saliva samples collected using the unstimulated collection method correlated with plasma levels. DHEA collected using the salivette device did not correlate significantly with either plasma or the unstimulated saliva (r 0.2;R(2) 0.04). It is crucial that future studies are aware of these issues and are cognizant of the effects of the method of collection when examining steroid levels in saliva.

  17. [Comparative analysis of dependence of saliva sorbitol and fructosamine levels on blood glucose level in patients with diabetes].

    Science.gov (United States)

    Morenkova, S A

    2004-01-01

    The possibility of determination of sorbitol and fructosamine in saliva has been studied in healthy volunteers and patients with diabetes. The dependence of these metabolites levels in saliva on blood glucose level was demonstrated. It is concluded that saliva sorbitol and fructosamine levels measurements may be used as diagnostic tests in diabetes and serve as indicators of efficacy of therapy in diabetes.

  18. A comparison of the effects of added saliva, α-amylase and water on texture perception in semisolids

    NARCIS (Netherlands)

    Engelen, L.; Wijk, R.A. de; Prinz, J.F.; Janssen, A.M.; Bilt, A. van der; Weenen, H.; Bosman, F.

    2003-01-01

    The effect of adding saliva or a saliva-related fluid (α-amylase solution and water) to custard prior to ingestion on the sensory ratings of odour, flavour and lip-tooth-, mouth- and after-feel sensations was investigated. Saliva had previously been collected from the subjects and each subject

  19. Friction between different wire bracket combinations in artificial saliva – an in vitro evaluation

    Science.gov (United States)

    FIDALGO, Tatiana Kelly da Silva; PITHON, Matheus Melo; MACIEL, José Vinicius Bolognesi; BOLOGNESE, Ana Maria

    2011-01-01

    Objective The objective this work was to assess the friction coefficient between brackets and wires of different materials under conditions simulating the oral environment. Material and Methods Stainless steel (SS) and titanium-molybdenum alloy (TMA) wires of 0.019x0.025-in diameter (American Orthodontics) and polycarbonate bracket (American Orthodontics), ceramic bracket (American Orthodontics), and metal bracket (3M Unitek) with slots of 0.022x0.030-in were used. The friction coefficient was assessed by means of mechanical traction with the system immersed in artificial saliva. The mean roughness of both wire surface and bracket slots was evaluated by using a surface profilometer. Results The system using TMA wire and polycarbonate bracket had the highest roughness (p<0.05). SS wire with ceramic bracket had the highest friction coefficient, whereas the use of metallic bracket yielded the lowest (p<0.05). However, it was observed a statistically significant difference in the system using TMA wire and ceramic bracket compared to that using TMA wire and polycarbonate bracket (p=0.038). Conclusion Ceramic brackets in association with SS wire should be judiciously used, since this system showed a high friction coefficient. PMID:21437471

  20. Cloning of a salivary gland metalloprotease and characterization of gelatinase and fibrin(ogen)lytic activities in the saliva of the Lyme Disease tick vector Ixodes scapularis

    OpenAIRE

    Francischetti, Ivo M. B.; Mather, Thomas N.; Ribeiro, José M. C.

    2003-01-01

    The full-length sequence of tick salivary gland cDNA coding for a protein similar to metalloproteases (MP) of the reprolysin family is reported. The Ixodes scapularis MP is a 488 aminoacid (aa) protein containing pre- and pro-enzyme domains, the zinc-binding motif HExxHxxGxxH common to metalloproteases and a cysteine-rich region. In addition, the predicted amino-terminal sequences of I. scapularis MPs were found by Edman degradation of PVDF-transferred SDS/PAGE-separated tick saliva proteins,...

  1. Saliva microbiomes distinguish caries-active from healthy human populations

    Science.gov (United States)

    Yang, Fang; Zeng, Xiaowei; Ning, Kang; Liu, Kuan-Liang; Lo, Chien-Chi; Wang, Wei; Chen, Jie; Wang, Dongmei; Huang, Ranran; Chang, Xingzhi; Chain, Patrick S; Xie, Gary; Ling, Junqi; Xu, Jian

    2012-01-01

    The etiology of dental caries remains elusive because of our limited understanding of the complex oral microbiomes. The current methodologies have been limited by insufficient depth and breadth of microbial sampling, paucity of data for diseased hosts particularly at the population level, inconsistency of sampled sites and the inability to distinguish the underlying microbial factors. By cross-validating 16S rRNA gene amplicon-based and whole-genome-based deep-sequencing technologies, we report the most in-depth, comprehensive and collaborated view to date of the adult saliva microbiomes in pilot populations of 19 caries-active and 26 healthy human hosts. We found that: first, saliva microbiomes in human population were featured by a vast phylogenetic diversity yet a minimal organismal core; second, caries microbiomes were significantly more variable in community structure whereas the healthy ones were relatively conserved; third, abundance changes of certain taxa such as overabundance of Prevotella Genus distinguished caries microbiota from healthy ones, and furthermore, caries-active and normal individuals carried different arrays of Prevotella species; and finally, no ‘caries-specific' operational taxonomic units (OTUs) were detected, yet 147 OTUs were ‘caries associated', that is, differentially distributed yet present in both healthy and caries-active populations. These findings underscored the necessity of species- and strain-level resolution for caries prognosis, and were consistent with the ecological hypothesis where the shifts in community structure, instead of the presence or absence of particular groups of microbes, underlie the cariogenesis. PMID:21716312

  2. Biochemical and Immunological Modifications in Saliva of SFINCSS Experiment

    Science.gov (United States)

    Volozhin, A. I.; Kuznetsov, P. A.; Ilyin, V. K.; Kuzmina, E. M.; Sashkina, T. I.

    of Russian and foreign volunteers and was divided onto 3 parts, 4 persons per each depending on isolation time. All the individuals were isolated days in confined habitat.: 1st group - 240 days; 2nd and 3rd - 110 days each. 1 group members were individually orally instructed on perfect dental care, 2nd group members were given an instruction how to use means for mouth and dental care. 3rd group was only studied but was not given any instruction. Biochemical studies of non-stimulated mixed saliva were done before and after the experiment. protein concentration increased due to increasing of it's density. The urea concentration did not changed. The glucose concentration changes were flexible within norm values before experiment and sufficiently increased after the experiment only in two individuals. Natrium and potassium level was stable and did not differed from normal value before and after the experiment. There was a tendency of decreasing of calcium concentration in volunteers saliva as a result of their long-term isolation. Concentration of non-organic phosphor did not changed. Alanintranspherase (ALT) activity increased 2-3 times in 3 volunteers, aspartataminotranspherase (AST) activity increased in three people. No changes were revealed for alpha-amilase. Content of IgG increased which fact indirectly suggest bacterial overgrowth. No changes in IgA and SIgA were estimated. of urea and glucose didn't changed. The concentration of calcium had a tendency to decrease, no changes for non-organic phosphor, potassium and natrium. However ALT and AST values sufficiently increased as well as IgG concentration. isolation, despite of individual measures of mouth and dental care, and in group of 110-day isolation with no hygienic measures. Significant indices of mouth and dental state in long-term isolation are levels of: protein ALT, AST (cytoplasmatic enzymes), and IgG.

  3. Prostaglandin E2 in tick saliva regulates macrophage cell migration and cytokine profile

    Science.gov (United States)

    2013-01-01

    Background Ticks are obligate hematophagous ectoparasites that suppress the host’s immune and inflammatory responses by secreting immuno-modulatory and anti-inflammatory molecules in their saliva. In previous studies we have shown that tick salivary gland extract (SGE) and saliva from Dermacentor variabilis have distinct effects on platelet-derived growth factor (PDGF)-stimulated IC-21 macrophage and NIH3T3-L1 fibroblast migration. Since tick saliva contains a high concentration of prostaglandin E2 (PGE2), a potent modulator of inflammation, we used a PGE2 receptor antagonist to evaluate the role of PGE2 in the different migratory responses induced by saliva and its impact on macrophage cytokine profile. Methods Adult ticks were fed on female New Zealand white rabbits for 5-8 days. Female ticks were stimulated with dopamine/theophylline to induce salivation and saliva was pooled. Competitive enzyme immunoassays (EIA) were used to measure saliva PGE2 content and the changes in macrophage intracellular cyclic adenosine monophosphate (cAMP) levels. The effects of tick saliva on macrophage and fibroblast migration were assessed in the absence and presence of the PGE2 receptor antagonist, AH 6809, using blind well chamber assays. A cytokine antibody array was used to examine the effects of tick saliva on macrophage cytokine secretion. Statistical significance was determined by one-way ANOVA; Student Newman-Kuels post-test was used for multiple comparisons. Results The saliva-induced increase in PDGF-stimulated macrophage migration was reversed by AH 6809. The inhibition of PDGF-stimulated fibroblast migration by saliva was also antagonist-sensitive. Tick saliva induced macrophages to secrete copious amounts of PGE2, and conditioned medium from these cells caused an AH 6809-sensitive inhibition of stimulated fibroblast migration, showing that macrophages can regulate fibroblast activity. We show that tick saliva decreased the secretion of the pro

  4. Yield stress fluids slowly yield to analysis

    NARCIS (Netherlands)

    Bonn, D.; Denn, M.M.

    2009-01-01

    We are surrounded in everyday life by yield stress fluids: materials that behave as solids under small stresses but flow like liquids beyond a critical stress. For example, paint must flow under the brush, but remain fixed in a vertical film despite the force of gravity. Food products (such as

  5. DNA decontamination of fingerprint brushes.

    Science.gov (United States)

    Szkuta, Bianca; Oorschot, Roland A H van; Ballantyne, Kaye N

    2017-08-01

    Genetic profiling of DNA collected from fingerprints that have been exposed to various enhancement techniques is routine in many forensic laboratories. As a result of direct contact with fingermark residues during treatment, there is concern around the DNA contamination risk of dusting fingermarks with fingerprint brushes. Previous studies have demonstrated the potential for cross-contamination between evidentiary items through various mechanisms, highlighting the risk of using the same fingerprint brush to powder multiple surfaces within and between crime-scenes. Experiments were performed to assess the contamination risk of reused fingerprint brushes through the transfer of dried saliva and skin deposits from and to glass surfaces with new unused squirrel hair and fiberglass brushes. Additional new unused brushes and brushes previously used in casework were also tested for their ability to contaminate samples. In addition, the ability to eradicate DNA from used squirrel hair and fiberglass fingerprint brushes was assessed using a 1% sodium hypochlorite solution and a 5% solution of a commercially available alternative, Virkon. DNA profiling results from surfaces contacted by treated and untreated brushes were compared to determine the effectiveness of the devised cleaning protocol. Brush durability was also assessed over multiple wash/rinse/dry cycles with both agents. Varying amounts of DNA-containing material were collected and transferred by squirrel hair and fiberglass brushes, with detectability on the secondary surface dependent on the biological nature of the material being transferred. The impact of DNA contamination from dirty fingerprint brushes was most apparent in simulations involving the transfer of dried saliva and brushes previously used in casework, while minimal transfer of touch DNA was observed. Alarmingly, large quantities of DNA were found to reside on new unused squirrel hair brushes, while no DNA was detected on new unused fiberglass

  6. Determination of acetaldehyde in saliva by gas-diffusion flow injection analysis.

    Science.gov (United States)

    Ramdzan, Adlin N; Mornane, Patrick J; McCullough, Michael J; Mazurek, Waldemar; Kolev, Spas D

    2013-07-05

    The consumption of ethanol is known to increase the likelihood of oral cancer. In addition, there has been a growing concern about possible association between long term use of ethanol-containing mouthwashes and oral cancer. Acetaldehyde, known to be a carcinogen, is the first metabolite of ethanol and it can be produced in the oral cavity after consumption or exposure to ethanol. This paper reports on the development of a gas-diffusion flow injection method for the online determination of salivary acetaldehyde by its colour reaction with 3-methyl-2-benzothiazolinone hydrazone (MBTH) and ferric chloride. Acetaldehyde samples and standards (80 μL) were injected into the donor stream containing NaCl from which acetaldehyde diffused through the hydrophobic Teflon membrane of the gas-diffusion cell into the acceptor stream containing the two reagents mentioned above. The resultant intense green coloured dye was monitored spectrophotometrically at 600 nm. Under the optimum working conditions the method is characterized by a sampling rate of 9h(-1), a linear calibration range of 0.5-15 mg L(-1) (absorbance=5.40×10(-2) [acetaldehyde, mg L(-1)], R(2)=0.998), a relative standard deviation (RSD) of 1.90% (n=10, acetaldehyde concentration of 2.5 mg L(-1)), and a limit of detection (LOD) of 12.3 μg L(-1). The LOD and sampling rate of the proposed method are superior to those of the conventional gas chromatographic (GC) method (LOD=93.0 μg L(-1) and sampling rate=4 h(-1)). The reliability of the proposed method was illustrated by the fact that spiked with acetaldehyde saliva samples yielded excellent recoveries (96.6-101.9%), comparable to those obtained by GC (96.4-102.3%) and there was no statistically significant difference at the 95% confidence level between the two methods when non-spiked saliva samples were analysed. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Performance of cryptococcal antigen lateral flow assay using saliva in Ugandans with CD4 <100.

    Directory of Open Access Journals (Sweden)

    Richard Kwizera

    Full Text Available Cryptococcal meningitis can best be diagnosed by cerebrospinal fluid India ink microscopy, cryptococcal antigen detection, or culture. These require invasive lumbar punctures. The utility of cryptococcal antigen detection in saliva is unknown. We evaluated the diagnostic performance of the point-of-care cryptococcal antigen lateral flow assay (CrAg LFA in saliva.We screened HIV-infected, antiretroviral therapy naïve persons with symptomatic meningitis (n = 130 and asymptomatic persons with CD4+<100 cells/µL entering into HIV care (n = 399 in Kampala, Uganda. The diagnostic performance of testing saliva was compared to serum/plasma cryptococcal antigen as the reference standard.The saliva lateral flow assay performance was overall more sensitive in symptomatic patients (88% than in asymptomatic patients (27%. The specificity of saliva lateral flow assay was excellent at 97.8% in the symptomatic patients and 100% in asymptomatic patients. The degree of accuracy of saliva in diagnosing cryptococcosis and the level of agreement between the two sample types was better in symptomatic patients (C-statistic 92.9, κ-0.82 than in asymptomatic patients (C-statistic 63.5, κ-0.41. Persons with false negative salvia CrAg tests had lower levels of peripheral blood CrAg titers (P<0.001.There was poor diagnostic performance in testing saliva for cryptococcal antigen, particularly among asymptomatic persons screened for preemptive treatment of cryptococcosis.

  8. 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.

  9. Performance of cryptococcal antigen lateral flow assay using saliva in Ugandans with CD4 <100.

    Science.gov (United States)

    Kwizera, Richard; Nguna, Joyce; Kiragga, Agnes; Nakavuma, Jesca; Rajasingham, Radha; Boulware, David R; Meya, David B

    2014-01-01

    Cryptococcal meningitis can best be diagnosed by cerebrospinal fluid India ink microscopy, cryptococcal antigen detection, or culture. These require invasive lumbar punctures. The utility of cryptococcal antigen detection in saliva is unknown. We evaluated the diagnostic performance of the point-of-care cryptococcal antigen lateral flow assay (CrAg LFA) in saliva. We screened HIV-infected, antiretroviral therapy naïve persons with symptomatic meningitis (n = 130) and asymptomatic persons with CD4+diagnostic performance of testing saliva was compared to serum/plasma cryptococcal antigen as the reference standard. The saliva lateral flow assay performance was overall more sensitive in symptomatic patients (88%) than in asymptomatic patients (27%). The specificity of saliva lateral flow assay was excellent at 97.8% in the symptomatic patients and 100% in asymptomatic patients. The degree of accuracy of saliva in diagnosing cryptococcosis and the level of agreement between the two sample types was better in symptomatic patients (C-statistic 92.9, κ-0.82) than in asymptomatic patients (C-statistic 63.5, κ-0.41). Persons with false negative salvia CrAg tests had lower levels of peripheral blood CrAg titers (Pdiagnostic performance in testing saliva for cryptococcal antigen, particularly among asymptomatic persons screened for preemptive treatment of cryptococcosis.

  10. NT-ProBNP levels in saliva and its clinical relevance to heart failure.

    Science.gov (United States)

    Foo, Jared Yong Yang; Wan, Yunxia; Kostner, Karam; Arivalagan, Alicia; Atherton, John; Cooper-White, Justin; Dimeski, Goce; Punyadeera, Chamindie

    2012-01-01

    Current blood based diagnostic assays to detect heart failure (HF) have large intra-individual and inter-individual variations which have made it difficult to determine whether the changes in the analyte levels reflect an actual change in disease activity. Human saliva mirrors the body's health and well being and ∼20% of proteins that are present in blood are also found in saliva. Saliva has numerous advantages over blood as a diagnostic fluid which allows for a non-invasive, simple, and safe sample collection. The aim of our study was to develop an immunoassay to detect NT-proBNP in saliva and to determine if there is a correlation with blood levels. Saliva samples were collected from healthy volunteers (n = 40) who had no underlying heart conditions and HF patients (n = 45) at rest. Samples were stored at -80°C until analysis. A customised homogeneous sandwich AlphaLISA((R)) immunoassay was used to quantify NT-proBNP levels in saliva. Our NT-proBNP immunoassay was validated against a commercial Roche assay on plasma samples collected from HF patients (n = 37) and the correlation was r(2) = 0.78 (pdiagnostic accuracy of 90.6%. We have firstly demonstrated that NT-proBNP can be detected in saliva and that the levels were higher in heart failure patients compared with healthy control subjects. Further studies will be needed to demonstrate the clinical relevance of salivary NT-proBNP in unselected, previously undiagnosed populations.

  11. Effect of masticatory stimulation on the quantity and quality of saliva and the salivary metabolomic profile.

    Science.gov (United States)

    Okuma, Nobuyuki; Saita, Makiko; Hoshi, Noriyuki; Soga, Tomoyoshi; Tomita, Masaru; Sugimoto, Masahiro; Kimoto, Katsuhiko

    2017-01-01

    This study characterized the changes in quality and quantity of saliva, and changes in the salivary metabolomic profile, to understand the effects of masticatory stimulation. Stimulated and unstimulated saliva samples were collected from 55 subjects and salivary hydrophilic metabolites were comprehensively quantified using capillary electrophoresis-time-of-flight mass spectrometry. In total, 137 metabolites were identified and quantified. The concentrations of 44 metabolites in stimulated saliva were significantly higher than those in unstimulated saliva. Pathway analysis identified the upregulation of the urea cycle and synthesis and degradation pathways of glycine, serine, cysteine and threonine in stimulated saliva. A principal component analysis revealed that the effect of masticatory stimulation on salivary metabolomic profiles was less dependent on sample population sex, age, and smoking. The concentrations of only 1 metabolite in unstimulated saliva, and of 3 metabolites stimulated saliva, showed significant correlation with salivary secretion volume, indicating that the salivary metabolomic profile and salivary secretion volume were independent factors. Masticatory stimulation affected not only salivary secretion volume, but also metabolite concentration patterns. A low correlation between the secretion volume and these patterns supports the conclusion that the salivary metabolomic profile may be a new indicator to characterize masticatory stimulation.

  12. Saliva DHEA and cortisol responses following short-term corticosteroid intake.

    Science.gov (United States)

    Jollin, L; Thomasson, R; Le Panse, B; Baillot, A; Vibarel-Rebot, N; Lecoq, A M; Amiot, V; De Ceaurriz, J; Collomp, K

    2010-02-01

    Given the high correlation between the serum and saliva hormone values demonstrated at rest, saliva provides a convenient non-invasive way to determine dehydroepiandrosterone (DHEA) and cortisol concentrations. However, to our knowledge, pituitary adrenal recovery following short-term suppression with corticosteroids has never been investigated in saliva. The aim of this study was therefore to examine how steroid hormone concentrations in saliva are influenced by short-term corticosteroid administration. We studied saliva DHEA and cortisol concentrations before, during (day 1-day 7) and following (day 8-day 16) the administration of oral therapeutic doses of prednisone (50 mg daily for 1 week) in 11 healthy recreationally trained women. Mean saliva DHEA and cortisol concentrations decreased immediately after the start of prednisone treatment (P DHEA and cortisol had returned to pretreatment levels. These data are consistent with previous studies on blood samples and suggest that non-invasive saliva samples may offer a practical approach to assessing pituitary-adrenal function continuously during and after short-term corticosteroid therapy.

  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. Ixodes scapularis saliva mitigates inflammatory cytokine secretion during Anaplasma phagocytophilum stimulation of immune cells

    Directory of Open Access Journals (Sweden)

    Chen Gang

    2012-10-01

    Full Text Available Abstract Background Ixodes scapularis saliva enables the transmission of infectious agents to the mammalian host due to its immunomodulatory, anesthetic and anti-coagulant properties. However, how I. scapularis saliva influences host cytokine secretion in the presence of the obligate intracellular rickettsial pathogen Anaplasma phagocytophilum remains elusive. Methods Bone marrow derived macrophages (BMDMs were stimulated with pathogen associated molecular patterns (PAMPs and A. phagocytophilum. Cytokine secretion was measured in the presence and absence of I. scapularis saliva. Human peripheral blood mononuclear cells (PBMCs were also stimulated with Tumor Necrosis Factor (TNF-α in the presence and absence of I. scapularis saliva and interleukin (IL-8 was measured. Results I. scapularis saliva inhibits inflammatory cytokine secretion by macrophages during stimulation of Toll-like (TLR and Nod-like receptor (NLR signaling pathways. The effect of I. scapularis saliva on immune cells is not restricted to murine macrophages because decreasing levels of interleukin (IL-8 were observed after TNF-α stimulation of human peripheral blood mononuclear cells. I. scapularis saliva also mitigates pro-inflammatory cytokine response by murine macrophages during challenge with A. phagocytophilum. Conclusions These findings suggest that I. scapularis may inhibit inflammatory cytokine secretion during rickettsial transmission at the vector-host interface.

  15. Effect of masticatory stimulation on the quantity and quality of saliva and the salivary metabolomic profile.

    Directory of Open Access Journals (Sweden)

    Nobuyuki Okuma

    Full Text Available This study characterized the changes in quality and quantity of saliva, and changes in the salivary metabolomic profile, to understand the effects of masticatory stimulation.Stimulated and unstimulated saliva samples were collected from 55 subjects and salivary hydrophilic metabolites were comprehensively quantified using capillary electrophoresis-time-of-flight mass spectrometry.In total, 137 metabolites were identified and quantified. The concentrations of 44 metabolites in stimulated saliva were significantly higher than those in unstimulated saliva. Pathway analysis identified the upregulation of the urea cycle and synthesis and degradation pathways of glycine, serine, cysteine and threonine in stimulated saliva. A principal component analysis revealed that the effect of masticatory stimulation on salivary metabolomic profiles was less dependent on sample population sex, age, and smoking. The concentrations of only 1 metabolite in unstimulated saliva, and of 3 metabolites stimulated saliva, showed significant correlation with salivary secretion volume, indicating that the salivary metabolomic profile and salivary secretion volume were independent factors.Masticatory stimulation affected not only salivary secretion volume, but also metabolite concentration patterns. A low correlation between the secretion volume and these patterns supports the conclusion that the salivary metabolomic profile may be a new indicator to characterize masticatory stimulation.

  16. Yield Improvement in Steel Casting (Yield II)

    Energy Technology Data Exchange (ETDEWEB)

    Richard A. Hardin; Christoph Beckermann; Tim Hays

    2002-02-18

    This report presents work conducted on the following main projects tasks undertaken in the Yield Improvement in Steel Casting research program: Improvement of Conventional Feeding and Risering Methods, Use of Unconventional Yield Improvement Techniques, and Case Studies in Yield Improvement. Casting trials were conducted and then simulated using the precise casting conditions as recorded by the participating SFSA foundries. These results present a statistically meaningful set of experimental data on soundness versus feeding length. Comparisons between these casting trials and casting trials performed more than forty years ago by Pellini and the SFSA are quite good and appear reasonable. Comparisons between the current SFSA feeding rules and feeding rules based on the minimum Niyama criterion reveal that the Niyama-based rules are generally less conservative. The niyama-based rules also agree better with both the trials presented here, and the casting trails performed by Pellini an d the SFSA years ago. Furthermore, the use of the Niyama criterion to predict centerline shrinkage for horizontally fed plate sections has a theoretical basis according to the casting literature reviewed here. These results strongly support the use of improved feeding rules for horizontal plate sections based on the Niyama criterion, which can be tailored to the casting conditions for a given alloy and to a desired level of soundness. The reliability and repeatability of ASTM shrinkage x-ray ratings was investigated in a statistical study performed on 128 x-rays, each of which were rated seven different times. A manual ''Feeding and Risering Guidelines for Steel Castings' is given in this final report. Results of casting trials performed to test unconventional techniques for improving casting yield are presented. These use a stacked arrangement of castings and riser pressurization to increase the casting yield. Riser pressurization was demonstrated to feed a casting up to

  17. Bond yield curve construction

    Directory of Open Access Journals (Sweden)

    Kožul Nataša

    2014-01-01

    Full Text Available In the broadest sense, yield curve indicates the market's view of the evolution of interest rates over time. However, given that cost of borrowing it closely linked to creditworthiness (ability to repay, different yield curves will apply to different currencies, market sectors, or even individual issuers. As government borrowing is indicative of interest rate levels available to other market players in a particular country, and considering that bond issuance still remains the dominant form of sovereign debt, this paper describes yield curve construction using bonds. The relationship between zero-coupon yield, par yield and yield to maturity is given and their usage in determining curve discount factors is described. Their usage in deriving forward rates and pricing related derivative instruments is also discussed.

  18. Saliva Proteins of Vector Culicoides Modify Structure and Infectivity of Bluetongue Virus Particles

    Science.gov (United States)

    Darpel, Karin E.; Langner, Kathrin F. A.; Nimtz, Manfred; Anthony, Simon J.; Brownlie, Joe; Takamatsu, Haru-Hisa; Mellor, Philip S.; Mertens, Peter P. C.

    2011-01-01

    Bluetongue virus (BTV) and epizootic haemorrhagic disease virus (EHDV) are related orbiviruses, transmitted between their ruminant hosts primarily by certain haematophagous midge vectors (Culicoides spp.). The larger of the BTV outer-capsid proteins, ‘VP2’, can be cleaved by proteases (including trypsin or chymotrypsin), forming infectious subviral particles (ISVP) which have enhanced infectivity for adult Culicoides, or KC cells (a cell-line derived from C. sonorensis). We demonstrate that VP2 present on purified virus particles from 3 different BTV strains can also be cleaved by treatment with saliva from adult Culicoides. The saliva proteins from C. sonorensis (a competent BTV vector), cleaved BTV-VP2 more efficiently than those from C. nubeculosus (a less competent / non-vector species). Electrophoresis and mass spectrometry identified a trypsin-like protease in C. sonorensis saliva, which was significantly reduced or absent from C. nubeculosus saliva. Incubating purified BTV-1 with C. sonorensis saliva proteins also increased their infectivity for KC cells ∼10 fold, while infectivity for BHK cells was reduced by 2–6 fold. Treatment of an ‘eastern’ strain of EHDV-2 with saliva proteins of either C. sonorensis or C. nubeculosus cleaved VP2, but a ‘western’ strain of EHDV-2 remained unmodified. These results indicate that temperature, strain of virus and protein composition of Culicoides saliva (particularly its protease content which is dependent upon vector species), can all play a significant role in the efficiency of VP2 cleavage, influencing virus infectivity. Saliva of several other arthropod species has previously been shown to increase transmission, infectivity and virulence of certain arboviruses, by modulating and/or suppressing the mammalian immune response. The findings presented here, however, demonstrate a novel mechanism by which proteases in Culicoides saliva can also directly modify the orbivirus particle structure, leading to

  19. The functions of human saliva: A review sponsored by the World Workshop on Oral Medicine VI.

    Science.gov (United States)

    Dawes, C; Pedersen, A M L; Villa, A; Ekström, J; Proctor, G B; Vissink, A; Aframian, D; McGowan, R; Aliko, A; Narayana, N; Sia, Y W; Joshi, R K; Jensen, S B; Kerr, A R; Wolff, A

    2015-06-01

    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 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 bolus, swallowing and speaking. Saliva provides the fluid in which solid tastants may dissolve and distributes tastants around the mouth to the locations of the taste buds. The hypotonic unstimulated saliva facilitates taste recognition. Salivary amylase is involved in digestion of starches. Saliva acts as a buffer to protect oral, pharyngeal and oesophageal mucosae from orally ingested acid or acid regurgitated from the stomach. Saliva protects the teeth against acid by contributing to the acquired enamel pellicle, which forms a renewable lubricant between opposing tooth surfaces, by being supersaturated with respect to tooth mineral, by containing bicarbonate as a buffer and urea and by facilitating clearance of acidic materials from the mouth. Saliva contains many antibacterial, antiviral and antifungal agents which modulate the oral microbial flora in different ways. Saliva also facilitates the healing of oral wounds. Clearly, saliva has many functions which are needed for proper protection and functioning of the human body. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Saliva proteins of vector Culicoides modify structure and infectivity of bluetongue virus particles.

    Directory of Open Access Journals (Sweden)

    Karin E Darpel

    2011-03-01

    Full Text Available Bluetongue virus (BTV and epizootic haemorrhagic disease virus (EHDV are related orbiviruses, transmitted between their ruminant hosts primarily by certain haematophagous midge vectors (Culicoides spp.. The larger of the BTV outer-capsid proteins, 'VP2', can be cleaved by proteases (including trypsin or chymotrypsin, forming infectious subviral particles (ISVP which have enhanced infectivity for adult Culicoides, or KC cells (a cell-line derived from C. sonorensis. We demonstrate that VP2 present on purified virus particles from 3 different BTV strains can also be cleaved by treatment with saliva from adult Culicoides. The saliva proteins from C. sonorensis (a competent BTV vector, cleaved BTV-VP2 more efficiently than those from C. nubeculosus (a less competent/non-vector species. Electrophoresis and mass spectrometry identified a trypsin-like protease in C. sonorensis saliva, which was significantly reduced or absent from C. nubeculosus saliva. Incubating purified BTV-1 with C. sonorensis saliva proteins also increased their infectivity for KC cells ∼10 fold, while infectivity for BHK cells was reduced by 2-6 fold. Treatment of an 'eastern' strain of EHDV-2 with saliva proteins of either C. sonorensis or C. nubeculosus cleaved VP2, but a 'western' strain of EHDV-2 remained unmodified. These results indicate that temperature, strain of virus and protein composition of Culicoides saliva (particularly its protease content which is dependent upon vector species, can all play a significant role in the efficiency of VP2 cleavage, influencing virus infectivity. Saliva of several other arthropod species has previously been shown to increase transmission, infectivity and virulence of certain arboviruses, by modulating and/or suppressing the mammalian immune response. The findings presented here, however, demonstrate a novel mechanism by which proteases in Culicoides saliva can also directly modify the orbivirus particle structure, leading to

  1. Chemokine expression of oral fibroblasts and epithelial cells in response to artificial saliva.

    Science.gov (United States)

    Müller, Heinz-Dieter; Cvikl, Barbara; Lussi, Adrian; Gruber, Reinhard

    2016-06-01

    Artificial saliva is widely used to overcome reduced natural salivary flow. Natural saliva provokes the expression of chemokines in oral fibroblasts in vitro. However, if artificial saliva changes the expression of chemokines remains unknown. Here, we investigated the ability of Saliva Orthana®, Aldiamed®, Glandosane®, and Saliva Natura® to change the expression of chemokines in human oral fibroblasts and the human oral epithelial cell line HSC-2 by means of reverse transcription polymerase chain reaction and immunoassays. Mucins isolated from bovine submaxillary glands and recombinant human mucin 1 were included in the bioassay. Formazan formation and LIVE/DEAD® staining determined the impact of artificial saliva on cell viability. The involvement of signaling pathways was determined by pharmacologic inhibitors and Western blotting. In gingival fibroblasts, Saliva Orthana®-containing mucins provoked a significantly increased expression of CXC ligand 8 (CXCL8, or interleukin 8), CXCL1, and CXCL2. Immunoassays for CXCL8 and CXCL1 confirmed the translation at the protein level. The respective dilution of artificial saliva had no impact on formazan formation and LIVE/DEAD® staining. Mucins isolated from bovine submaxillary glands also increased the panel of chemokine expression in gingival fibroblasts. BAY 11-7082, a nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) inhibitor, but also TAK-242, an inhibitor of toll-like receptor 4 signaling, blocked chemokine expression of Saliva Orthana® and bovine mucins. In HSC-2 cells, Glandosane® significantly increased CXCL8 expression. Saliva Orthana® stimulated chemokine expression in gingival fibroblasts. Mammalian mucins, but also possible contaminations with endotoxins, might contribute to the respective changes in gene expression. Epithelial cells have a differential response to artificial saliva with Glandosane® changing CXCL8 expression. Artificial saliva can incite a cellular response

  2. 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...

  3. Analysis of methamphetamine in hair, nail, sweat, and saliva by mass fragmentography.

    Science.gov (United States)

    Suzuki, S; Inoue, T; Hori, H; Inayama, S

    1989-01-01

    A method for the detection and quantitation of methamphetamine and its major metabolite in hair, nails, sweat, and saliva from habitual users of methamphetamine by mass fragmentography has been developed. Hair and nail samples were washed with water and methanol to remove the external contamination, processed with 0.6M HCl, alkalinized, and extracted with CHCl3/isopropanol (3:1 v/v). Sweat and saliva samples were extracted with methanol. After trifluoroacetyl derivatization, the samples were analyzed by mass fragmentography. Methamphetamine and its major metabolite, amphetamine, were detected in hair, nail, and sweat samples, but methamphetamine alone was detected in saliva samples.

  4. Supplementation of xylitol-containing chewing gum with probiotics: a double blind, randomised pilot study focusing on saliva flow and saliva properties.

    Science.gov (United States)

    Gueimonde, Laura; Vesterlund, Satu; García-Pola, María J; Gueimonde, Miguel; Söderling, Eva; Salminen, Seppo

    2016-03-01

    The aim of this study was to investigate the impact of daily chewing, for 12 weeks, of 2 different probiotic gums compared with placebo on saliva flow rate, saliva IgA levels and saliva pH. The intervention study included 54 adult volunteers with hyposalivation in a double-blind, randomised and placebo-controlled design with three parallel groups. Volunteers were randomly assigned to 3 different groups: subjects in group A (n = 19) were given placebo chewing gum, group B (n = 17) received Bifidobacterium animalis ssp. lactis Bb12 (ATCC 27536) and group C (n = 18) received Lactobacillus rhamnosus LGG (ATCC 53103), Bifidobacterium longum 46 (DSM 14583) and Bifidobacterium longum 2C (DSM 14579) gums, during 3 months. Two volunteers from group B left the study for personal reasons leaving 19, 15 and 18 volunteers, respectively, for analyses. Clinical examinations, personal interviews, sialometries and saliva sampling were conducted at baseline and after 1, 2, 3 and 4 months. No statistically significant differences were found between probiotic and placebo groups for any of the parameters analysed. No side effects of probiotic or placebo chewing gums were observed. Chewing gum, with and without probiotics, had a positive impact on salivary flow rate and saliva pH and IgA levels.

  5. The use of liquid chromatography tandem mass spectrometry to detect proteins in saliva from horses with and without systemic inflammation

    DEFF Research Database (Denmark)

    Jacobsen, Stine; Top Adler, Ditte Marie; Bundgaard, Louise

    2014-01-01

    The objective of the study was to assess global expression of proteins in equine saliva using liquid chromatography tandem mass spectrometry (LC-MS/MS). Saliva was obtained from seven horses with and six horses without evidence of systemic inflammatory disease. Tryptic peptides from saliva were...... analysed by LC-MS/MS. Of 195 unique proteins identified, 57 were detected only in saliva samples from horses with systemic inflammation (in two to six of the seven horses). Among the differentially expressed proteins were several acute phase proteins (APPs) such as serum amyloid A, fibrinogen, haptoglobin......, and alpha1-acid glycoprotein. The study is the first to describe detection of inflammatory proteins in horse saliva. The proteins detected were similar to those described in saliva from cattle, small ruminants and pigs. Detection of APPs in horses with systemic inflammation suggests that saliva may be used...

  6. [An experimental study on PAc and GTF gene vaccines of Streptococcus mutans against rats caries: antibody levels in saliva and serum].

    Science.gov (United States)

    Yang, Deqin; Liu, Tianjia; Cao, Fuxian

    2003-10-01

    The purpose of this study is to examine the levels of salivary SIgA and serum IgG induced by pcDNA3-pac and pcDNA3-gtfB immunization, so as to testify the antigenity of the two gene vaccines. 36 28-day-old Wistar rats were divided into 6 groups, among which 3 experimental groups were vaccinated with pcDNA3-pac, pcDNA3-gtfB or pcDNA3-pac combined with pcDNA3-gtfB, respectively, one positive control was vaccinated with inactive whole cell of S. mutans JBP and other two negative controls were injected with the vector pcDNA3 or PBS buffer, respectively. All vaccines and materials were delivered with 100 micrograms by submandibular gland injection for 3 times. Then the restricted bacterial model of rat was constructed. Following that all rats were fed with cariogenic diet Keyes 2000 for 3 months, saliva and serum samples were collected to assay SIgA or IgG levels by ELASA. The salivary S-IgA levels both in pcDNA3-pac combined with pcDNA3-gtfB group and inactive S. mutans cell group were higher than others (P vaccine and inactive S. mutans vaccination reached its peak at the 11th week after the first inoculation and kept until the end of the study. Both pcDNA3-pac and pcDNA3-gtfB can express immunogenic protein and induce immune responses of mucosal and humoral immune system in gnobobiotic rats. It is also indicated that the joint gene vaccines immunization is an optimal choice for anticaries strategy.

  7. 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

  8. Saliva characteristics, diet and carioreceptivity in dental students.

    Science.gov (United States)

    Chifor, Ioana; Badea, Iulia; Chifor, Radu; Popa, Dan; Staniste, Liviu; Tarmure, Dragos; Avram, Ramona

    2014-01-01

    The use of sugar by dental plaque microorganisms leads to acid formation from the bacteria metabolism, which determines a decrease of pH onto teeth surfaces. The value of the critical pH is 5.2-5.5. We aimed to evaluate the capacity of patients to change their diet towards caries prevention after acknowledging the values of saliva parameters (pH, buffer capacity). A group of 52 subjects were clinically examined according to the International Caries Assessment and Detection System protocol. They were required to complete a diet questionnaire and salivary tests were made for the oral mucosa hydration level, pH, buffer capacity, salivary flow rate at rest and upon stimulation. 4 pre-calibrated 6th year students and 2 dentists performed the tests and the ICDAS examination. One week after the tests, the subjects were asked to complete the diet questionnaire again. The studied group consisted of students aged between 23-26 years, randomly selected among 6(th) year students of the Faculty of Dentistry from Cluj-Napoca. The mean DMF-S index was 18.39. Most of the patients (65%) had a DMF-S index between 9 and 21. Just 2.5% had an index of 3, which was the lowest value recorded. 5% of the patients had a DMFS of 35, which was the maximal value recorded. The distribution of DMF-S was normal. 50% of the patients had no active caries. Even though most subjects (19.23%) had a pH within the normal interval, most of them were at the bottom value of the interval (6.8). Most subjects had a pH of 6.4, which is moderately acid. The mean pH was 6.7, therefore, a moderately acid one. The Pearson correlation coefficient between DMFS and pH was 0.255. A mild negative correlation (-0.275) was found between the cariogenic food and buffer capacity. A week later we noticed a statistically significant decrease of cariogenic foods and drinks in students with acid pH and with low buffer capacity. A regular intake of cakes, bonbons and chocolate was reported by subjects who had a high DMF-S value

  9. 49 CFR 40.263 - What happens when an employee is unable to provide a sufficient amount of saliva for an alcohol...

    Science.gov (United States)

    2010-10-01

    ... a sufficient amount of saliva for an alcohol screening test? 40.263 Section 40.263 Transportation... sufficient amount of saliva for an alcohol screening test? (a) As the STT, you must take the following steps if an employee is unable to provide sufficient saliva to complete a test on a saliva screening device...

  10. Microbial profile comparisons of saliva, pooled and site-specific subgingival samples in periodontitis patients

    DEFF Research Database (Denmark)

    Belstrøm, Daniel; Sembler-Møller, Maria Lynn; Grande, Maria Anastasia

    2017-01-01

    . DESIGN: Site specific subgingival plaque samples (n = 54), pooled subgingival plaque samples (n = 18) and stimulated saliva samples (n = 18) were collected from 18 patients with generalized chronic periodontitis. Subgingival and salivary microbiotas were characterized by means of HOMINGS (Human Oral......OBJECTIVES: The purpose of this study was to compare microbial profiles of saliva, pooled and site-specific subgingival samples in patients with periodontitis. We tested the hypotheses that saliva can be an alternative to pooled subgingival samples, when screening for presence of periopathogens...... by pooled subgingival samples. Presence of Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, Filifactor alocis, Tannerella forsythia and Parvimona micra in site-specific subgingival samples were detected in saliva with an AUC of 0.79 (sensitivity: 0.61, specificity: 0.94), compared...

  11. Alkaline phosphatase levels in patients with coronary heart disease saliva and its relation with periodontal status

    Science.gov (United States)

    Yunita, Dina Suci; Masulili, Sri Lelyati C.; Tadjoedin, Fatimah M.; Radi, Basuni

    2017-02-01

    Coronary heart disease (CHD) is a disease that causes narrowing of the coronary arteries. Currently, there is a hypothesis regarding periodontal infection that increases risk for heart disease. Alkaline phosphatase (ALP) as a marker of inflammation will increase in atherosclerosis and periodontal disease. The objective of this research is analyzing the relationship between the levels of alkaline phosphatase in saliva with periodontal status in patients with CHD and non CHD. Here, saliva of 104 subjects were taken, each 1 ml, and levels of Alkaline Phosphatase was analyzed using Abbott ci4100 architect. We found that no significant difference of Alkaline Phosphatase levels in saliva between CHD patients and non CHD. Therefore, it can be concluded that Alkaline Phosphatase levels in patients with CHD saliva was higher than non CHD and no association between ALP levels with periodontal status.

  12. Saliva in studies of epidemiology of human disease: the UK Biobank project.

    Science.gov (United States)

    Galloway, John W; Keijser, Bart J F; Williams, David M

    2016-02-01

    There has been immense interest in the uses of saliva in the diagnosis of systemic disease over the past decade and longer because it is recognized that saliva possesses great potential as a diagnostic fluid. In spite of this, the usefulness of saliva in studies of the epidemiology of human disease has still to be properly evaluated. This review describes the UK Biobank project and explores the scope to use this and other such cohort studies to gain important insights into the epidemiological aspects of systemic disease. The Biobank holds around 85,000 well-characterized saliva samples, together with blood and urine samples, the results of a battery of physiological tests, a full medical history and a detailed description of the subject's lifestyle. This repository is a resource for insightful and highly powered oral and dental research. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  13. Saliva as a diagnostic fluid in sports medicine: potential and limitations

    National Research Council Canada - National Science Library

    Nunes, Lázaro Alessandro Soares; Macedo, Denise Vaz de

    2013-01-01

    ... mostly a less invasive method in comparison with venous blood collection. The saliva is a hypotonic fluid in relation to plasma, containing compounds produced in the salivary glands (immunoglobulin A [IgA] and α-amylase...

  14. [Evaluation of functional adaptation level in air specialists according to biochemical indexes of saliva secretion].

    Science.gov (United States)

    Soldatov, S K; Malysheva, E V; Zasiad'ko, K I; Abashev, V Iu; Gulin, A V; Ermakova, N V

    2009-09-01

    It was examined a capability of evaluation of functional condition of air staff by indexes of natrium, kalium, cortisol and glucose in saliva. There were realized 5 series of examinations with participations of 71 airplane pilot of the same level in conditions of realizing flies of different difficultness. Saliva sampling was effectuated before and after the flies not later then 10-15 minutes after landing. On pre-flight medical examination and after performance of task of air relay there was registration of systolic, diasystolic blood pressure and cardiac rate. It was posed the correlation of physiological indexes with percentage of examined ingredients in saliva in different flight loads. The results of examinations speak for capability of using of indexes of percentage of natrium, kalium, cortisol and glucose in saliva for evaluation of functional condition of airplane pilots during effectuating the flies and rating of value of flight load with account of individual peculiarities.

  15. Stability of unstimulated and stimulated whole saliva flow rates in children.

    Science.gov (United States)

    Sánchez-Pérez, Leonor; Irigoyen-Camacho, Esther; Sáenz-Martínez, Laura; Zepeda Zepeda, Marco; Acosta-Gío, Enrique; Méndez-Ramírez, Ignacio

    2016-09-01

    To analyze the stability of the unstimulated saliva flow rate (USFR) and the stimulated saliva flow rate (SSFR) in children followed from age 7 to 12 years old. Longitudinal study. Whole saliva samples were collected from school children (50 girls and 50 boys). Forty-four girls and 32 boys remained in this cohort for 6 years (dropout rate 24%). Variables that could influence USFR or SSFR patterns were analyzed in a repeated-measures manova. Over a 6-year follow-up, the children's USFR ranged from 0.41 to 0.46 mL/min in the initial and final observation, respectively, and showed no significant differences (P = 0.4455) during the follow-up. The children consistently belonged to one of three distinct SSFR groups (P saliva for screening or diagnostic purposes. © 2015 BSPD, IAPD and John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  16. Alpha-2-HS-glycoprotein (AHSG) polymorphism in semen and saliva.

    Science.gov (United States)

    Yasuda, T; Takeshita, H; Tsubota, E; Sawazaki, K; Iida, R; Nadano, D; Kishi, K

    1996-04-01

    Polymorphism of alpha-2-HS-glycoprotein (AHSG) was demonstrated in human semen and whole saliva samples by thin-layer polyacrylamide gel isoelectric focusing (IEF) and immunoblotting. Although the seminal AHSG IEF patterns were found to differ from those of plasma AHSG from the corresponding donors, incorporation of Nonidet P-40 into the IEF gel (pH 4.2-4.9) enabled us to phenotype seminal AHSG correctly. Salivary AHSG, however, exhibited IEF patterns similar to those of the corresponding plasma AHSG. By treating the samples with neuraminidase, it was possible to determine the AHSG types using 2-5 microL semen and 50-100 microL whole saliva samples. The AHSG types determined separately in 47 sets of semen, whole saliva, urine and plasma samples from the same donors correlated perfectly with each other. AHSG typing could, therefore, provide an additional discriminant characteristic in the forensic examination of semen and saliva samples.

  17. Saliva collection by using filter paper for measuring cortisol levels in dogs.

    Science.gov (United States)

    Oyama, D; Hyodo, M; Doi, H; Kurachi, T; Takata, M; Koyama, S; Satoh, T; Watanabe, G

    2014-01-01

    Four experiments were conducted to evaluate the accuracy and reliability of noninvasive evaluation of cortisol in saliva of dogs. In experiment 1, we measured the cortisol concentration in the filter paper on which 250-μL cortisol solutions had been quantitatively pipetted and in filter papers dipped in cortisol solution. In experiment 2, we collected the blood and saliva of dogs 3 times at 30-min intervals and compared the cortisol concentrations to examine whether the dynamics of cortisol in the blood and saliva are similar. The results of experiments 1 and 2 showed that the cortisol concentration can be quantitatively measured with this method and that the dynamics of cortisol concentration in the plasma and saliva collected by using filter paper are not different (P = 0.14 for experiment 1 and P = 0.51 for experiment 2). In experiment 3, to investigate the factors related to inducing stress in dogs by using the filter-paper method of collecting saliva, we compared the cortisol concentrations at 0 and 30 min after collecting the saliva of pet dogs. The dog owners completed a survey on their dogs, providing basic information and reporting the collection of their dog's saliva. We found that the cortisol concentrations increased significantly in dogs whose owners spent >2 min collecting saliva (P = 0.005), suggesting that prompt collection of saliva is necessary for accurate assessment of cortisol without induction of a stress response. In addition, the cortisol concentrations increased significantly in dogs whose teeth were not regularly brushed (P = 0.04), suggesting that regular teeth brushing mitigates the effect of the collection process on cortisol concentrations in the saliva, with minimal stress to the dogs. In experiment 4, we measured cortisol concentrations in pet dogs accustomed to having their teeth brushed by their owners, before and after interaction with their owners, to assess whether brushing induces stress in dogs. We detected that the

  18. Zika virus infection spread through saliva – a truth or myth?

    Directory of Open Access Journals (Sweden)

    Walter Luiz SIQUEIRA

    2016-01-01

    Full Text Available Abstract In this Point-of-view article we highlighted some features related to saliva and virus infection, in special for zika virus. In addition, we pointed out the potential oral problems caused by a microcephaly originated by a zika virus infection. In the end the, we demonstrated the importance of a more comprehensive exploration of saliva and their components as a fluid for diagnostic and therapeutic approaches on oral and systemic diseases.

  19. A Simple Saliva-Based Test for Detecting Antibodies to Human Immunodeficiency Virus*

    OpenAIRE

    Schramm, Willfried; Angulo, Gustavo Barriga; Torres, Patricia Castillo; Burgess-Cassler, Anthony

    1999-01-01

    This study was performed to determine the feasibility of using saliva as a diagnostic medium for the detection of antibodies to human immunodeficiency virus type 1 (HIV-1) and HIV-2 under nonlaboratory conditions and to evaluate the performance characteristics of such a test. We developed for this purpose a self-contained kit (Saliva · Strip [ST]), which combines the collection and processing, as well as the analysis, of the specimen. The kit’s performance was eval...

  20. Comparative analysis of bacterial profiles in unstimulated and stimulated saliva samples

    DEFF Research Database (Denmark)

    Belstrøm, Daniel; Holmstrup, Palle; Jensen, Allan Bardow

    2016-01-01

    BACKGROUND AND OBJECTIVE: The microbial profiles of stimulated saliva samples have been shown to differentiate between patients with periodontitis, patients with dental caries, and orally healthy individuals. Saliva was stimulated to allow for easy and rapid collection; however, microbial...... orally and systemically healthy, non-smoking participants. Salivary bacterial profiles were analyzed by means of the Human Oral Microbe Identification using Next Generation Sequencing (HOMINGS), and statistical analysis was performed using Mann-Whitney test with Benjamini-Hochberg's correction...

  1. Zika virus infection spread through saliva--a truth or myth?

    Science.gov (United States)

    Siqueira, Walter Luiz; Moffa, Eduardo Buozi; Mussi, Maria Carolina Martins; Machado, Maria Aparecida de Andrade Moreira

    2016-01-01

    In this Point-of-view article we highlighted some features related to saliva and virus infection, in special for zika virus. In addition, we pointed out the potential oral problems caused by a microcephaly originated by a zika virus infection. In the end the, we demonstrated the importance of a more comprehensive exploration of saliva and their components as a fluid for diagnostic and therapeutic approaches on oral and systemic diseases.

  2. Clinical and diagnostic utility of saliva as a non-invasive diagnostic fluid: a systematic review

    OpenAIRE

    Nunes, Lazaro Alessandro Soares; MUSSAVIRA, Sayeeda; Bindhu, Omana Sukumaran

    2015-01-01

    This systematic review presents the latest trends in salivary research and its applications in health and disease. Among the large number of analytes present in saliva, many are affected by diverse physiological and pathological conditions. Further, the non-invasive, easy and cost-effective collection methods prompt an interest in evaluating its diagnostic or prognostic utility. Accumulating data over the past two decades indicates towards the possible utility of saliva to monitor overall hea...

  3. Effect of human saliva on the fluoride sensitivity of glucose uptake by Streptococcus mutans.

    Science.gov (United States)

    Germaine, G R; Tellefson, L M

    1981-01-01

    The fluoride (F) sensitivity of glucose uptake by whole cell suspensions of streptococcus mutans in the presence and absence of human whole salivary supernatant was studied. It was observed that dithiothreitol (DTT) and other thiols markedly reduced the F sensitivity of cells when saliva (50%, vol/vol) was present during glucose uptake. In the absence of saliva, cells were sensitive to 2 to 2.5 mM F regardless of the presence of thiols. Supplementation of cells in phosphate or tris(hydroxymethyl)aminomethane-hydrochloride buffers with physiological concentrations of calcium or phosphate had no effect on the F sensitivity of the organism. Experiments with permeabilized cells suggested that thiols themselves had no direct effect on the F sensitivity of enolase (a principal F target). Cells pretreated with DDT subsequently exhibited decreased F sensitivity when examined in the presence of saliva but not in the absence of saliva. Cells pretreated with whole salivary supernatant were found to be subsequently less sensitive to F in the absence of saliva during glucose uptake. Furthermore, in cases where cells were pretreated with saliva, subsequent additions of DDT were unnecessary to obtain maximal reduction in the F sensitivity of glucose uptake. It was concluded that the saliva-dependent reduction in F sensitivity of glucose uptake was not due to sequestration of available F by salivary constituents. The data suggest that a salivary component(s) interacts directly with the microorganism in some manner which results in reduced F sensitivity of the process under study. Possible mechanisms of saliva action are discussed. PMID:7333673

  4. 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.

  5. Saliva of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) inhibits classical and alternative complement pathways.

    Science.gov (United States)

    Silva, Naylene C S; Vale, Vladimir F; Franco, Paula F; Gontijo, Nelder F; Valenzuela, Jesus G; Pereira, Marcos H; Sant'Anna, Mauricio R V; Rodrigues, Daniel S; Lima, Walter S; Fux, Blima; Araujo, Ricardo N

    2016-08-11

    Rhipicephalus (Boophilus) microplus is the main ectoparasite affecting livestock worldwide. For a successful parasitism, ticks need to evade several immune responses of their hosts, including the activation of the complement system. In spite of the importance of R. microplus, previous work only identified one salivary molecule that blocks the complement system. The current study describes complement inhibitory activities induced by R. microplus salivary components and mechanisms elicited by putative salivary proteins on both classical and alternative complement pathways. We found that R. microplus saliva from fully- and partially engorged females was able to inhibit both pathways. Saliva acts strongly at the initial steps of both complement activation pathways. In the classical pathway, the saliva blocked C4 cleavage, and hence, deposition of C4b on the activation surface, suggesting that the inhibition occurs at some point between C1q and C4. In the alternative pathway, saliva acts by binding to initial components of the cascade (C3b and properdin) thereby preventing the C3 convertase formation and reducing C3b production and deposition as well as cleavage of factor B. Saliva has no effect on formation or decay of the C6 to C8 components of the membrane attack complex. The saliva of R. microplus is able to inhibit the early steps of classical and alternative pathways of the complement system. Saliva acts by blocking C4 cleavage and deposition of C4b on the classical pathway activation surface and, in the alternative pathway, saliva bind to initial components of the cascade (C3b and properdin) thereby preventing the C3 convertase formation and the production and deposition of additional C3b.

  6. Investigation of Fe and Ca in non-stimulated human saliva using NAA

    Science.gov (United States)

    de Medeiros, J. A. G.; Zamboni, C. B.; Kovacs, L.; Lewgoy, H. R.

    2015-07-01

    In this study we investigated non-stimulated human whole saliva of healthy subjects and patients with periodontal disease using Neutron Activation Analysis technique (NAA). The measurements were performed in the IEA-R1 nuclear reactor at IPEN-CNEN/SP. We found considerable metabolic changes mainly in Fe and Ca concentration in whole saliva of periodontal patients. These data are useful for identifying or preventing this oral disease in the Brazilian population.

  7. Genome-Wide Identification of Genes Essential for the Survival of Streptococcus pneumoniae in Human Saliva

    Science.gov (United States)

    Verhagen, Lilly M.; de Jonge, Marien I.; Burghout, Peter; Schraa, Kiki; Spagnuolo, Lorenza; Mennens, Svenja; Eleveld, Marc J.; van der Gaast-de Jongh, Christa E.; Zomer, Aldert; Hermans, Peter W. M.; Bootsma, Hester J.

    2014-01-01

    Since Streptococcus pneumoniae transmits through droplet spread, this respiratory tract pathogen may be able to survive in saliva. Here, we show that saliva supports survival of clinically relevant S. pneumoniae strains for more than 24 h in a capsule-independent manner. Moreover, saliva induced growth of S. pneumoniae in growth-permissive conditions, suggesting that S. pneumoniae is well adapted for uptake of nutrients from this bodily fluid. By using Tn-seq, a method for genome-wide negative selection screening, we identified 147 genes potentially required for growth and survival of S. pneumoniae in saliva, among which genes predicted to be involved in cell envelope biosynthesis, cell transport, amino acid metabolism, and stress response predominated. The Tn-seq findings were validated by testing a panel of directed gene deletion mutants for their ability to survive in saliva under two testing conditions: at room temperature without CO2, representing transmission, and at 37°C with CO2, representing in-host carriage. These validation experiments confirmed that the plsX gene and the amiACDEF and aroDEBC operons, involved in respectively fatty acid metabolism, oligopeptide transport, and biosynthesis of aromatic amino acids play an important role in the growth and survival of S. pneumoniae in saliva at 37°C. In conclusion, this study shows that S. pneumoniae is well-adapted for growth and survival in human saliva and provides a genome-wide list of genes potentially involved in adaptation. This notion supports earlier evidence that S. pneumoniae can use human saliva as a vector for transmission. PMID:24586856

  8. A Study on Duration of Effect of Transcutaneous Electrical Nerve Stimulation Therapy on Whole Saliva Flow.

    Science.gov (United States)

    Bhasin, Neha; Reddy, Sreedevi; Nagarajappa, Anil Kumar; Kakkad, Ankur

    2015-06-01

    Saliva is a complex fluid, whose important role is to maintain the well being of oral cavity. Salivary gland hypofunction or hyposalivation is the condition of having reduced saliva production which leads to the subjective complaint of oral dryness termed xerostomia.(7) Management of xerostomia includes palliative therapy using topical agents or systemic therapy. Electrostimulation to produce saliva was studied in the past and showed moderate promise but never became part of mainstream therapy. Hence, this study was undertaken to evaluate the effect of transcutaneous electrical nerve stimulation (TENS) on whole salivary flow rate in healthy adults and to evaluate how long this effect of TENS lasts on salivary flow. One hundred healthy adult subjects were divided into five age groups with each group containing 20 subjects equally divided into males and females in each group. Unstimulated saliva was collected using a graduated test tube fitted with funnel and quantity was measured. Transcutaneous electrical nerve stimulation unit was activated and stimulated saliva was collected. Saliva was again collected 30 minutes and 24 hours post stimulation. The mean unstimulated whole saliva flow rate for all subjects (n = 100) was 2.60 ml/5 min. During stimulation, it increased to 3.60 ± 0.39 ml/5 min. There was 38.46% increase in salivary flow. Ninety six out of 100 responded positively to TENS therapy. Salivary flow remained increased 30 minutes and 24 hours post stimulation with the values being 3.23 ± 0.41 ml/5 min and 2.69 ± 0.39 ml/5 min respectively. Repeated measures One way analysis of variance (ANOVA) test showed that the difference between these values were statistically significant. Transcutaneous electrical nerve stimulation therapy was effective for stimulation of whole saliva in normal, healthy subjects and its effect retained till 30 minutes and a little up to 24 hours. Transcutaneous electrical nerve stimulation may work best synergistically with other

  9. Salivary IgA in minor-gland saliva of children, adolescents, and young adults.

    Science.gov (United States)

    Sonesson, Mikael; Hamberg, Kristina; Wallengren, Marie-Louise Lundin; Matsson, Lars; Ericson, Dan

    2011-02-01

    According to previous studies, minor glands produce about 35% of the total salivary immunoglobulin A (salivary IgA). The age-dependent increase in whole-saliva salivary IgA concentrations has been studied extensively, but we found no published reports comparing the minor-gland saliva concentrations of salivary IgA in children, adolescents, and adults. In this study we measured the concentration of salivary IgA in saliva from the labial and the buccal minor glands of children, adolescents, and adults. Three age groups donated saliva for analysis: 3-yr-old children, 14-yr-old adolescents, and 20- to 25-yr-old adults. Minor-gland saliva was collected on filter paper and unstimulated whole saliva was collected by draining into a tube, and the salivary IgA concentration was determined by ELISA. The salivary IgA concentration in labial saliva was significantly lower among 3-yr-old children (0.037 mg 100 ml(-1), SD = 0.035) than among 14-yr-old adolescents (0.126 mg 100 ml(-1), SD = 0.128) and adults (0.128 mg 100 ml(-1), SD = 0.13). The 3-yr-old children also had significantly lower whole-saliva salivary IgA values compared with the other age groups (0.09 mg 100 ml(-1), SD = 0.091; 0.179 mg 100 ml(-1), SD = 0.149; and 0.170 mg 100 ml(-1), SD = 0.099, respectively). This increase in salivary IgA concentrations with age might reflect a developing immune response in the growing child. © 2011 Eur J Oral Sci.

  10. Screening of children saliva samples for bisphenol A using stochastic, amperometric and multimode microsensors

    OpenAIRE

    Stefan-Van Staden, Raluca-Ioana; Gugoaşă, Livia Alexandra; Calenic, Bogdan; van Staden, Jacobus F.; Legler, Juliette

    2014-01-01

    Bisphenol A found in plastic vessels used for children feeding is an endocrine disrupting compound. Therefore it can also induce the obesity at a very early stage in the life of children, and its presence in children saliva should be checked. We proposed eight microsensors: four stochastic microsensors, one amperometric microsensor and three multimode microsensors for the screening of children saliva for bisphenol A in a concentration range from 10−15 to 10−4 mol/L. Qualitative assessment of ...

  11. Evaluation of Innate Immune Biomarkers in Saliva for Diagnostic Potential of Bacterial and Viral Respiratory Infection

    Science.gov (United States)

    2014-02-03

    up-regulated in bacterial and viral infections, such as Staphylococcus aureus and Influenza virus, and plays a key role in cell-mediated immunity... Streptococcus sore throat compact pneumoniae 2 58 male white Saliva/serum Cough, fatigue, fever, Vitek 2 Streptococcus sore throat compact pneumoniae 3 52...fever, fatigue, Vitek 2 Streptococcus sore throat compact pneumoniae 8 44 female white Saliva/serum Cough, fever, muscle Vitek 2 Klebsiella

  12. Genome-wide identification of genes essential for the survival of Streptococcus pneumoniae in human saliva.

    Directory of Open Access Journals (Sweden)

    Lilly M Verhagen

    Full Text Available Since Streptococcus pneumoniae transmits through droplet spread, this respiratory tract pathogen may be able to survive in saliva. Here, we show that saliva supports survival of clinically relevant S. pneumoniae strains for more than 24 h in a capsule-independent manner. Moreover, saliva induced growth of S. pneumoniae in growth-permissive conditions, suggesting that S. pneumoniae is well adapted for uptake of nutrients from this bodily fluid. By using Tn-seq, a method for genome-wide negative selection screening, we identified 147 genes potentially required for growth and survival of S. pneumoniae in saliva, among which genes predicted to be involved in cell envelope biosynthesis, cell transport, amino acid metabolism, and stress response predominated. The Tn-seq findings were validated by testing a panel of directed gene deletion mutants for their ability to survive in saliva under two testing conditions: at room temperature without CO2, representing transmission, and at 37 °C with CO2, representing in-host carriage. These validation experiments confirmed that the plsX gene and the amiACDEF and aroDEBC operons, involved in respectively fatty acid metabolism, oligopeptide transport, and biosynthesis of aromatic amino acids play an important role in the growth and survival of S. pneumoniae in saliva at 37 °C. In conclusion, this study shows that S. pneumoniae is well-adapted for growth and survival in human saliva and provides a genome-wide list of genes potentially involved in adaptation. This notion supports earlier evidence that S. pneumoniae can use human saliva as a vector for transmission.

  13. In situ assessment of the saliva effect on enamel morphology after microabrasion technique

    OpenAIRE

    Pini,Núbia Inocencya Pavesi; Lima, Débora Alves Nunes Leite; Sundfeld,Renato Herman; Ambrosano,Gláucia Maria Bovi; Aguiar,Flávio Henrique Baggio; Lovadino,José Roberto

    2014-01-01

    AIM: This study evaluated saliva effects on enamel morphology surface after microabrasion technique. METHODS: Enamel blocks (16 mm2) obtained from bovine incisors were divided into 9 groups as follows: one control group (no treatment), four groups with microabrasion treatment using 35% phosphoric acid and pumice (H3PO4+Pum) and other four groups treated with 6.6% hydrochloric acid and silica (HCl+Sil). One group of each treatment was submitted to 4 frames of saliva exposure: without exposure,...

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

    African Journals Online (AJOL)

    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.

  15. Radioimmunoassay for progesterone in human saliva during the menstrual cycle

    Energy Technology Data Exchange (ETDEWEB)

    Luisi, M.; Franchi, F.; Kicovic, P.M.; Silvestri, D.; Cossu, G.; Catarsi, A.L.; Barletta, D.; Gasperi, M. (Pisa Univ. (Italy))

    1981-10-01

    A sensitive, specific and accurate radioimmunoassay of progesterone in human saliva is described, using /sup 3/H. The assay had a sensitivity of 8 pg/tube and blanks were negligible. The intra- and inter-assay coefficients of variation were 5.2 and 9.4%, respectively. The mean recovery from 60 samples was 93.2 +- 6.3%. Results obtained from nine healthy, normally menstruating women showed that salivary progesterone rose from the 4th day before ovulation to a mean peak (+- SD) of 1.14 +- 0.17 ng/ml on the 8th day after ovulation, followed by a gradual decline. Correlation of salivary and simultaneously obtained plasma progesterone levels was good (r = 0.47; P < 0.001), although the maximum percent increase in salivary progesterone was more than 10 times greater than that of plasma progesterone. Salivary progesterone is thought to reflect the unbound fraction of plasma progesterone and this non-invasive technique can be used for serial investigations in which frequent samplings are required.

  16. Corrosion of dental alloys in artificial saliva with Streptococcus mutans.

    Directory of Open Access Journals (Sweden)

    Chunhui Lu

    Full Text Available A comparative study of the corrosion resistance of CoCr and NiCr alloys in artificial saliva (AS containing tryptic soy broth (Solution 1 and Streptococcus mutans (S. mutans species (Solution 2 was performed by electrochemical methods, including open circuit potential measurements, impedance spectroscopy, and potentiodynamic polarization. The adherence of S. mutans to the NiCr and CoCr alloy surfaces immersed in Solution 2 for 24 h was verified by scanning electron microscopy, while the results of electrochemical impedance spectroscopy confirmed the importance of biofilm formation for the corrosion process. The R(QR equivalent circuit was successfully used to fit the data obtained for the AS mixture without S. mutans, while the R(Q(R(QR circuit was found to be more suitable for describing the biofilm properties after treatment with the AS containing S. mutans species. In addition, a negative shift of the open circuit potential with immersion time was observed for all samples regardless of the solution type. Both alloys exhibited higher charge transfer resistance after treatment with Solution 2, and lower corrosion current densities were detected for all samples in the presence of S. mutans. The obtained results suggest that the biofilm formation observed after 24 h of exposure to S. mutans bacteria might enhance the corrosion resistance of the studied samples by creating physical barriers that prevented oxygen interactions with the metal surfaces.

  17. 210Po in Human Saliva of Smokeless Tobacco Users.

    Science.gov (United States)

    Meli, Maria Assunta; Desideri, Donatella; Roselli, Carla; Feduzi, Laura

    2017-01-01

    The occurrence and mobility of Po in oral smokeless tobacco products (STPs) were determined because its effects on human health must be taken into account. This research was subdivided into two parts: determination by alpha spectrometry of the Po activity concentration in 16 oral smokeless tobacco products of different brands purchased in local specialty stores in Europe and evaluation of its percent extraction into an artificial salivary gland during sucking or chewing operations. Polonium-210 was detected in all samples, and its concentrations ranged from 3.46 to 14.8 Bq kg (mean value of 7.45 ± 3.82 Bq kg). The highest concentration was found in chewing tobacco. The samples showed no significant difference in the content of Po level. The data obtained in this study show that the polonium, although poorly extracted (12.8 ± 8.96%) by artificial saliva, is not totally retained within the smokeless tobacco products, with a consequent potential health hazard associated with oral use of these products.

  18. Corrosion of dental alloys in artificial saliva with Streptococcus mutans.

    Science.gov (United States)

    Lu, Chunhui; Zheng, Yuanli; Zhong, Qun

    2017-01-01

    A comparative study of the corrosion resistance of CoCr and NiCr alloys in artificial saliva (AS) containing tryptic soy broth (Solution 1) and Streptococcus mutans (S. mutans) species (Solution 2) was performed by electrochemical methods, including open circuit potential measurements, impedance spectroscopy, and potentiodynamic polarization. The adherence of S. mutans to the NiCr and CoCr alloy surfaces immersed in Solution 2 for 24 h was verified by scanning electron microscopy, while the results of electrochemical impedance spectroscopy confirmed the importance of biofilm formation for the corrosion process. The R(QR) equivalent circuit was successfully used to fit the data obtained for the AS mixture without S. mutans, while the R(Q(R(QR))) circuit was found to be more suitable for describing the biofilm properties after treatment with the AS containing S. mutans species. In addition, a negative shift of the open circuit potential with immersion time was observed for all samples regardless of the solution type. Both alloys exhibited higher charge transfer resistance after treatment with Solution 2, and lower corrosion current densities were detected for all samples in the presence of S. mutans. The obtained results suggest that the biofilm formation observed after 24 h of exposure to S. mutans bacteria might enhance the corrosion resistance of the studied samples by creating physical barriers that prevented oxygen interactions with the metal surfaces.

  19. Saliva levels of Abeta1-42 as potential biomarker of Alzheimer's disease: a pilot study.

    Science.gov (United States)

    Bermejo-Pareja, Felix; Antequera, Desiree; Vargas, Teo; Molina, Jose A; Carro, Eva

    2010-11-03

    Simple, non-invasive tests for early detection of degenerative dementia by use of biomarkers are urgently required. However, up to the present, no validated extracerebral diagnostic markers for the early diagnosis of Alzheimer disease (AD) are available. The clinical diagnosis of probable AD is made with around 90% accuracy using modern clinical, neuropsychological and imaging methods. A biochemical marker that would support the clinical diagnosis and distinguish AD from other causes of dementia would therefore be of great value as a screening test. A total of 126 samples were obtained from subjects with AD, and age-sex-matched controls. Additionally, 51 Parkinson's disease (PD) patients were used as an example of another neurodegenerative disorder. We analyzed saliva and plasma levels of β amyloid (Aβ) using a highly sensitive ELISA kit. We found a small but statistically significant increase in saliva Aβ42 levels in mild AD patients. In addition, there were not differences in saliva concentration of Aβ42 between patients with PD and healthy controls. Saliva Aβ40 expression was unchanged within all the studied sample. The association between saliva Aβ42 levels and AD was independent of established risk factors, including age or Apo E, but was dependent on sex and functional capacity. We suggest that saliva Aβ42 levels could be considered a potential peripheral marker of AD and help discrimination from other types of neurodegenerative disorders. We propose a new and promising biomarker for early AD.

  20. Pengaruh Kontaminsi Saliva terhadap Kekuatan Tarik antara Resin Komposit dengan Jaringan Dentin

    Directory of Open Access Journals (Sweden)

    Andi Soufyan

    2013-06-01

    Full Text Available Composite resin are restorative materials having color similar to teeth and have been widely used in dentistry. The successful application of composite resin influences the duration of the restoration in the oral cavity. The aim of this research is to describe the influence of artificial saliva contamination and the application of re-conditioning on tensile bond strength of composite resin to dentin. In the control group, the dentin were etched, bonding were applied and composite resin were restored on the dentin. In the group with artificial saliva contamination without re-conditioning, the dentin were etched, bonding were applied and then contaminated with artificial saliva, dried and then restired with composite resin. While the group with artificial saliva contamination with re-conditioning, the dentin were etched, bonding were applied and contaminated with artificial saliva, and then etched and applied bonding agent and restored composite resin.Bond strength test used “Universal testing machine, AG 5000. The results showed that highest value of tensile bond strength of composite resin to dentin was at the control group. It can be concluded that artificial saliva contamination decreased tensile bond strength while  re-conditioning application increased it.DOI: 10.14693/jdi.v15i2.69dentin

  1. Growth of Candida albicans in human saliva is supported by low-molecular-mass compounds.

    Science.gov (United States)

    Valentijn-Benz, Marianne; Nazmi, Kamran; Brand, Henk S; van't Hof, Wim; Veerman, Enno C I

    2015-12-01

    Saliva plays a key role in the maintenance of a stable oral microflora. It contains antimicrobial compounds but also functions as a substrate for growth of bacteria under conditions of low external nutrient supply. Besides bacteria, yeasts, in particular Candida albicans, commonly inhabit the oral cavity. Under immunocompromised conditions, instantaneous outgrowth of this yeast occurs in oral carriers of C. albicans, suggesting that this yeast is able to survive in the oral cavity with saliva as sole source of growth substrate. The aim of the present study was to identify the salivary constituents that are used by C. albicans for growth and survival in saliva. In addition, we have explored the effect of growth in saliva on the susceptibility of C. albicans to histatin 5, a salivary antifungal peptide. It was found that C. albicans was able to grow in human saliva without addition of glucose, and in the stationary phase could survive for more than 400 h. Candida albicans grown in saliva was more than 10 times less susceptible for salivary histatin 5 than C. albicans cultured in Sabouraud medium. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. Saliva levels of Abeta1-42 as potential biomarker of Alzheimer's disease: a pilot study

    Directory of Open Access Journals (Sweden)

    Antequera Desiree

    2010-11-01

    Full Text Available Abstract Background Simple, non-invasive tests for early detection of degenerative dementia by use of biomarkers are urgently required. However, up to the present, no validated extracerebral diagnostic markers for the early diagnosis of Alzheimer disease (AD are available. The clinical diagnosis of probable AD is made with around 90% accuracy using modern clinical, neuropsychological and imaging methods. A biochemical marker that would support the clinical diagnosis and distinguish AD from other causes of dementia would therefore be of great value as a screening test. A total of 126 samples were obtained from subjects with AD, and age-sex-matched controls. Additionally, 51 Parkinson's disease (PD patients were used as an example of another neurodegenerative disorder. We analyzed saliva and plasma levels of β amyloid (Aβ using a highly sensitive ELISA kit. Results We found a small but statistically significant increase in saliva Aβ42 levels in mild AD patients. In addition, there were not differences in saliva concentration of Aβ42 between patients with PD and healthy controls. Saliva Aβ40 expression was unchanged within all the studied sample. The association between saliva Aβ42 levels and AD was independent of established risk factors, including age or Apo E, but was dependent on sex and functional capacity. Conclusions We suggest that saliva Aβ42 levels could be considered a potential peripheral marker of AD and help discrimination from other types of neurodegenerative disorders. We propose a new and promising biomarker for early AD.

  3. Effects of isoflurane anesthesia and pilocarpine on rat parotid saliva flow

    DEFF Research Database (Denmark)

    Knudsen, Jacob Dronninglund; Nauntofte, Birgitte; Josipovic, M

    2011-01-01

    The purpose of this study was to investigate the effects of isoflurane on unstimulated and pilocarpine-stimulated parotid saliva secretion. Ten male Sprague-Dawley rats weighing 350-400 g were randomized into two groups, and the saliva flow rate and lag phase were measured at two doses of isoflur......The purpose of this study was to investigate the effects of isoflurane on unstimulated and pilocarpine-stimulated parotid saliva secretion. Ten male Sprague-Dawley rats weighing 350-400 g were randomized into two groups, and the