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Sample records for women recognize trypsin

  1. Antibodies from malaria-exposed pregnant women recognize trypsin resistant epitopes on the surface of Plasmodium falciparum-infected erythrocytes selected for adhesion to chondroitin sulphate A

    Directory of Open Access Journals (Sweden)

    Staalsoe Trine

    2004-09-01

    Full Text Available Abstract Background The ability of Plasmodium falciparum-infected erythrocytes to adhere to the microvasculature endothelium is thought to play a causal role in malaria pathogenesis. Cytoadhesion to endothelial receptors is generally found to be highly sensitive to trypsinization of the infected erythrocyte surface. However, several studies have found that parasite adhesion to placental receptors can be markedly less sensitive to trypsin. This study investigates whether chondroitin sulphate A (CSA binding parasites express trypsin-resistant variant surface antigens (VSA that bind female-specific antibodies induced as a result of pregnancy associated malaria (PAM. Methods Fluorescence activated cell sorting (FACS was used to measure the levels of adult Scottish and Ghanaian male, and Ghanaian pregnant female plasma immunoglobulin G (IgG that bind to the surface of infected erythrocytes. P. falciparum clone FCR3 cultures were used to assay surface IgG binding before and after selection of the parasite for adhesion to CSA. The effect of proteolytic digestion of parasite erythrocyte surface antigens on surface IgG binding and adhesion to CSA and hyaluronic acid (HA was also studied. Results P. falciparum infected erythrocytes selected for adhesion to CSA were found to express trypsin-resistant VSA that are the target of naturally acquired antibodies from pregnant women living in a malaria endemic region of Ghana. However in vitro adhesion to CSA and HA was relatively trypsin sensitive. An improved labelling technique for the detection of VSA expressed by CSA binding isolates has also been described. Conclusion The VSA expressed by CSA binding P. falciparum isolates are currently considered potential targets for a vaccine against PAM. This study identifies discordance between the trypsin sensitivity of CSA binding and surface recognition of CSA selected parasites by serum IgG from malaria exposed pregnant women. Thus, the complete molecular

  2. Domestic Violence against Women: Recognize Patterns, Seek Help

    Science.gov (United States)

    ... this sounds familiar, you might be experiencing domestic violence. Domestic violence — also called intimate partner violence — occurs between ... www.womenshealth.gov/violence-against-women/types-of-violence/domestic-intimate-partner-violence.html. Accessed Jan. 7, 2016. ...

  3. Making the invisible become visible: Recognizing women's relationship with technology

    OpenAIRE

    Corneliussen, Hilde G.

    2014-01-01

    This discussion paper explores a new rhetoric that might help to increase our understanding of women’s relationships with information and communication technology. We have often heard the claim that women have to give up part of their femininity in technological contexts. However, it is not always femininity women have to "give up", the author argues, but rather their close bond with technology – a "something else" that has no precise name, and which for that reason can slip away in an al...

  4. Domestic Violence against Women: Recognize Patterns, Seek Help

    Science.gov (United States)

    ... emotional, sexual and physical abuse and threats of abuse. Men are sometimes abused by partners, but domestic violence is most often directed toward women. Domestic violence can happen in heterosexual or same-sex relationships. Abusive relationships always involve an imbalance of ...

  5. Recognizing the Two Faces of Gambling: The Lived Experiences of Korean Women Gamblers.

    Science.gov (United States)

    Kim, Sungjae; Kim, Wooksoo; Dickerson, Suzanne S

    2016-10-01

    The aim of this study was to explore the lived experiences of women problem gamblers, focusing on the meaning of gambling to them, how and why these women continue to gamble or stop gambling, and their needs and concerns. In order to effectively help women problem gamblers, practical in-depth knowledge is necessary to develop intervention programs for prevention, treatment, and recovery among women problem gamblers. The hermeneutic phenomenology approach was used to guide in-depth interviews and team interpretation of data. Sixteen women gamblers who chose to live in the casino area were recruited through snowball sampling with help from a counseling center. Participants were individually interviewed from February to April 2013 and asked to tell their stories of gambling. Transcribed interviews provided data for interpretive analysis. In the study analysis one constitutive pattern was identified: moving beyond addiction by recognizing the two faces of gambling in their life. Four related themes emerged in the analysis-gambling as alluring; gambling as 'ugly'; living in contradictions; and moving beyond. Loneliness and isolation play a critical role in gambling experiences of women gamblers in Korea. In other words, they are motivated to gamble in order to escape from loneliness, to stop gambling for fear of being lonely as they get older, and to stay in the casnio area so as not to be alone. The need for acceptance is one fo the important factors that should be considered in developing intervention program for women.

  6. Trypsin inhibition by ferrocene

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    Kukrić Zoran Z.

    2005-01-01

    Full Text Available Many transition metals and their complexes show inhibitory effect on someproteolytic enzymes, including trypsin. Their inhibitory activity is based on the direct binding to the active site of trypsin, mimicking formation of a five-coordinate transition state required for the reaction. The influence of ferrocene on trypsin activity using N-α-benzoyl-DL-arginine p-nitroanilide as a substrate was investigated. Ferrocene was selected as a potential inhibitor because it belongs to the family oforganometallic, so called "sandwich " compounds, and its cyclopentadienyl rings might have an appropriate geometry. It was found that ferrocene decreases trypsin activity and the K. for ferrocene was found to be 39.8.

  7. Trypsin and chymotrypsin in stool

    Science.gov (United States)

    ... be seen in a stool sample. This article discusses the test to measure trypsin and chymotrypsin in stool. ... drop of stool is placed on a thin layer of gelatin. If trypsin or chymotrypsin are present, ...

  8. Social anxiety disorder women easily recognize fearfull, sad and happy faces: the influence of gender.

    Science.gov (United States)

    Arrais, Kátia C; Machado-de-Sousa, João Paulo; Trzesniak, Clarissa; Santos Filho, Alaor; Ferrari, Maria Cecília F; Osório, Flávia L; Loureiro, Sonia R; Nardi, Antonio E; Hetem, Luiz Alberto B; Zuardi, Antonio W; Hallak, Jaime Eduardo C; Crippa, José Alexandre S

    2010-06-01

    It has been suggested that individuals with social anxiety disorder (SAD) are exaggeratedly concerned about approval and disapproval by others. Therefore, we assessed the recognition of facial expressions by individuals with SAD, in an attempt to overcome the limitations of previous studies. The sample was formed by 231 individuals (78 SAD patients and 153 healthy controls). All individuals were treatment naïve, aged 18-30 years and with similar socioeconomic level. Participants judged which emotion (happiness, sadness, disgust, anger, fear, and surprise) was presented in the facial expression of stimuli displayed on a computer screen. The stimuli were manipulated in order to depict different emotional intensities, with the initial image being a neutral face (0%) and, as the individual moved on across images, the expressions increased their emotional intensity until reaching the total emotion (100%). The time, accuracy, and intensity necessary to perform judgments were evaluated. The groups did not show statistically significant differences in respect to the number of correct judgments or to the time necessary to respond. However, women with SAD required less emotional intensity to recognize faces displaying fear (p=0.002), sadness (p=0.033) and happiness (p=0.002), with no significant differences for the other emotions or men with SAD. The findings suggest that women with SAD are hypersensitive to threat-related and approval-related social cues. Future studies investigating the neural basis of the impaired processing of facial emotion in SAD using functional neuroimaging would be desirable and opportune. (c) 2009 Elsevier Ltd. All rights reserved.

  9. Fundamentalist Protestant Christian Women: Recognizing Cultural and Gender Influences on Domestic Violence.

    Science.gov (United States)

    Foss, Louisa L.; Warnke, Melanie A.

    2003-01-01

    Multicultural, family process and structure, and gender concepts are used to provide a framework for understanding supports for and barriers to mental health experienced by abused fundamentalist Protestant Christian (FPC) women. For FPC women who are victims of domestic violence, these factors may intersect to prohibit or facilitate healthy life…

  10. Fragile X premutation in women: recognizing the health challenges beyond primary ovarian insufficiency.

    Science.gov (United States)

    Hoyos, Luis R; Thakur, Mili

    2017-03-01

    Fragile X premutation carriers have 55-200 CGG repeats in the 5' untranslated region of the FMR1 gene. Women with this premutation face many physical and emotional challenges in their life. Approximately 20% of these women will develop fragile X-associated primary ovarian insufficiency (FXPOI). In addition, they suffer from increased rates of menstrual dysfunction, diminished ovarian reserve, reduction in age of menopause, infertility, dizygotic twinning, and risk of having an offspring with a premutation or full mutation. Consequent chronic hypoestrogenism may result in impaired bone health and increased cardiovascular risk. Neuropsychiatric issues include risk of developing fragile X-associated tremor/ataxia syndrome, neuropathy, musculoskeletal problems, increased prevalence of anxiety, depression, and sleep disturbances independent of the stress of raising an offspring with fragile X syndrome and higher risk of postpartum depression. Some studies have reported a higher prevalence of thyroid abnormalities and hypertension in these women. Reproductive health providers play an important role in the health supervision of women with fragile X premutation. Awareness of these risks and correlation of the various manifestations could help in early diagnosis and coordination of care and services for these women and their families. This paper reviews current evidence regarding the possible conditions that may present in women with premutation-sized repeats beyond FXPOI.

  11. Shelters for Battered Women and Their Children: An Under-Recognized Source of Communicable Disease Transmission.

    Science.gov (United States)

    Gross, Thomas P.; Rosenberg, Mark L.

    1987-01-01

    Reports on a study of various aspects of communicable disease occurrence (focusing on diarrheal illness) in shelters for battered women and their children, the extent and methods of screening for disease, training of staff in health care principles, resources available for health care needs, and the health regulations governing these shelters. (PS)

  12. Sleep disturbances and memory impairment among pregnant women consuming khat: An under-recognized problem

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    Md. Dilshad Manzar

    2017-01-01

    Full Text Available Khat (Catha edulis is a evergreen flowering shrub that is cultivated at high altitudes, especially in East Africa and the southwest of the Arabian Peninsula. The plant contains alkaloids, of which cathinone and cathine have structural similarity and pharmacological action similar to amphetamines. The leaves are, therefore, consumed in some regions as a psychoactive stimulant due to cultural beliefs and misperceptions on the health benefits of khat consumption. This resulted in a growing prevalence of khat consumption among pregnant women. The myriad of physiological changes associated with pregnancy impairs sleep and memory. Moreover, khat has also been shown to have adverse effects on memory and sleep. Therefore, its use during pregnancy may further aggravate those impairments. The purpose of this mini-review is to summarize the changes in sleep and memory during pregnancy and the evidence supporting a relationship between khat consumption and neurocognitive deficits and sleep dysfunctions. The misperceptions of beneficial effects of khat, the high prevalence of consumption among pregnant women, and the possibility of under-reporting of khat abuse do necessitate the development of alternative methodologies to identify cases of unreported khat abuse in pregnant women. It is proposed that screening for sleep problems and memory deficits may help identify under-reported cases of khat abuse in pregnant women.

  13. Laura: Soybean variety lacking Kunitz trypsin inhibitor

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    Srebrić Mirjana

    2010-01-01

    Full Text Available Grain of conventional soybean varieties requires heat processing to break down trypsin inhibitor's activity before using as food or animal feed. At the same time, protein denaturation and other qualitative changes occur in soybean grain, especially if the temperature of heating is not controlled. Two types of trypsin inhibitor were found in soybean grain the Kunitz trypsin inhibitor and the Bowman-Birk inhibitor. Mature grain of soybean Laura is lacking Kunitz trypsin inhibitor. Grain yield of variety Laura is equal to high yielding varieties from the maturity group I, where it belongs. Lacking of Kunitz-trypsin inhibitor makes soybean grain suitable for direct feeding in adult non ruminant animals without previous thermal processing. Grain of variety Laura can be processed for a shorter period of time than conventional soybeans. This way we save energy, and preserve valuable nutritional composition of soybean grain, which is of interest in industrial processing.

  14. A preliminary neutron crystallography on the trypsin-bovine pancreatic trypsin inhibitor complex

    Energy Technology Data Exchange (ETDEWEB)

    Kawamura, K; Yamada, T; Kurihara, K; Tamada, T; Kuroki, R; Tanaka, I; Takahashi, H; Niimura, N, E-mail: niimura@mx.ibaraki.ac.jp

    2010-11-01

    Trypsin is one of serine proteases. BPTI (Bovine Pancreatic Trypsin Inhibitor) is a protein inhibitor, which binds trypsin tightly and inhibits cleavage of peptide bonds. X-ray structure determination of trypsin-BPTI complex could make clear the overview of the active site. However, information of hydrogen atoms related to catalytic mechanism has not been satisfied. In this study, the trypsin-BPTI complex structure has been determined by neutron diffraction data at 2.0 A resolution. Deuterium atoms of catalytic triad, hydration structures in the binding pocket of trypsin and hydrogen bonds were observed. We would like to discuss details of hydrogen bonds in the interface between trypsin and BPTI and the adjacent water molecules including hydrogen atoms involved in the enzymatic reaction.

  15. The WHISK (Women's Health: Increasing the Awareness of Science and Knowledge) Pilot Project: Recognizing Sex and Gender Differences in Women's Health and Wellness

    OpenAIRE

    Edwards, Lorece V.; Dennis, Sabriya; Weaks, Francesca

    2013-01-01

    Women's health encompasses a continuum of biological, psychological, and social challenges that differ considerably from those of men. Despite the remarkable advances in science, women's health and sex differences research is slowly gaining recognition and acceptance. It is important that women's health gain attention as women are usually the gatekeepers of care for the family. Women's health and health outcomes are strongly influenced by sex and gender differences as well as geography. Around t...

  16. HYDROLYSIS OF CHEESEWHEY PROTEINSWITH TRYPSIN, CHYMOTRYPSINAND CARBOXYPEPTIDASEA

    Directory of Open Access Journals (Sweden)

    M. F. CUSTÓDIO

    2009-01-01

    Full Text Available

    This work presents a method for adding value to cheese whey residues by whey proteins hydrolysis, using trypsin, chymotrypsin and carboxypeptidase A as catalysts. Sweet cheese whey was dialyzed and filtered in kaolin. Lactose and protein contents were analyzed after each step. The activities of bovine pancreas trypsin and chymotrypsin were measured at different pHs and temperatures. The optimal pH for the hydrolysis of whey proteins was 9.0 for both enzymes. Optima temperatures were 60ºC for trypsin, and 50ºC for chymotrypsin. Trypsin exhibited typical Michaelis-Menten behavior, but chymotrypsin did not. Electrophoretic analysis showed that neither trypsin nor chymotrypsin alone hydrolyzed whey proteins in less than three hours. Hydrolysis rates of -lactalbumin by trypsin, and of bovine serum albumin by chymotrypsin were low. When these enzymes were combined, however, all protein fractions were attacked and rates of hydrolysis were enhanced by one order of magnitude. The addition of carboxypeptidase A to the others enzymes did not improve the process yield.

  17. Antibodies from malaria-exposed pregnant women recognize trypsin resistant epitopes on the surface of Plasmodium falciparum-infected erythrocytes selected for adhesion to chondroitin sulphate A

    DEFF Research Database (Denmark)

    Sharling, Lisa; Enevold, Anders; Sowa, Kordai M P

    2004-01-01

    . falciparum clone FCR3 cultures were used to assay surface IgG binding before and after selection of the parasite for adhesion to CSA. The effect of proteolytic digestion of parasite erythrocyte surface antigens on surface IgG binding and adhesion to CSA and hyaluronic acid (HA) was also studied. RESULTS: P...

  18. Labor Market Modeling Recognizing Latent Job Attributes and Opportunity Constraints An Empirical Analysis of Labor Market Behavior of Eritrean Women

    OpenAIRE

    Marie W. Arneberg; Dagsvik, John K.; Jia, Zhiyang

    2002-01-01

    Abstract: This paper analyzes labor market behavior of urban Eritrean women with particular reference to the impact of education, earnings and labor market opportunities. Unlike traditional models of labor supply, which assume that work can be supplied freely in the labor market, we develop a framework that explicitly takes into account the notion of job opportunities and observable sets of feasible jobs. The framework is formulated within a random utility setting in which unob...

  19. The WHISK (Women's Health: Increasing the Awareness of Science and Knowledge) Pilot Project: Recognizing Sex and Gender Differences in Women's Health and Wellness.

    Science.gov (United States)

    Edwards, Lorece V; Dennis, Sabriya; Weaks, Francesca

    2013-09-01

    Women's health encompasses a continuum of biological, psychological, and social challenges that differ considerably from those of men. Despite the remarkable advances in science, women's health and sex differences research is slowly gaining recognition and acceptance. It is important that women's health gain attention as women are usually the gatekeepers of care for the family. Women's health and health outcomes are strongly influenced by sex and gender differences as well as geography. Around the world, the interplay of biology and culture brings about differences in men's and women's health, which have been largely overlooked. The Women's Health: Increasing the Awareness of Science and Knowledge (WHISK) Pilot Project was a multidisciplinary project aimed to increase the awareness of sex and gender differences in women's health and research among healthcare professionals. Theater expression and creative art were used to translate knowledge, enhance understanding, and increase the awareness of sex differences. Findings from this project clearly showed an apparent increase in knowledge and cultivation of new insights.

  20. Preventing Unintended Pregnancy Among Young Sexually Active Women: Recognizing the Role of Violence, Self-Esteem, and Depressive Symptoms on Use of Contraception.

    Science.gov (United States)

    Nelson, Deborah B; Zhao, Huaqing; Corrado, Rachel; Mastrogiannnis, Dimitrios M; Lepore, Stephen J

    2017-04-01

    Ineffective contraceptive use among young sexually active women is extremely prevalent and poses a significant risk for unintended pregnancy (UP). Ineffective contraception involves the use of the withdrawal method or the inconsistent use of other types of contraception (i.e., condoms and birth control pills). This investigation examined violence exposure and psychological factors related to ineffective contraceptive use among young sexually active women. Young, nonpregnant sexually active women (n = 315) were recruited from an urban family planning clinic in 2013 to participate in a longitudinal study. Tablet-based surveys measured childhood violence, community-level violence, intimate partner violence, depressive symptoms, and self-esteem. Follow-up surveys measured type and consistency of contraception used 9 months later. Multivariate logistic regression models assessed violence and psychological risk factors as main effects and moderators related to ineffective compared with effective use of contraception. The multivariate logistic regression model showed that childhood sexual violence and low self-esteem were significantly related to ineffective use of contraception (adjusted odds ratio [aOR] = 2.69, confidence interval [95% CI]: 1.18-6.17, and aOR = 0.51, 95% CI: 0.28-0.93; respectively), although self-esteem did not moderate the relationship between childhood sexual violence and ineffective use of contraception (aOR = 0.38, 95% CI: 0.08-1.84). Depressive symptoms were not related to ineffective use of contraception in the multivariate model. Interventions to reduce UP should recognize the long-term effects of childhood sexual violence and address the role of low self-esteem on the ability of young sexually active women to effectively and consistently use contraception to prevent UP.

  1. Plasma antibodies from malaria-exposed pregnant women recognize variant surface antigens on Plasmodium falciparum-infected erythrocytes in a parity-dependent manner and block parasite adhesion to chondroitin sulfate A

    DEFF Research Database (Denmark)

    Ricke, C H; Staalsoe, T; Koram, K

    2000-01-01

    In areas of intense Plasmodium falciparum transmission, clinical immunity is acquired during childhood, and adults enjoy substantial protection against malaria. An exception to this rule is pregnant women, in whom malaria is both more prevalent and severe than in nonpregnant women. Pregnancy...... an area of hyperendemic P. falciparum transmission generally possessed low levels of Abs specifically recognizing surface Ags expressed by a CSA-adhering parasite isolate, while unselected isolates were well recognized. In marked contrast, most third-trimester pregnant women from that area had very high...

  2. Can You Recognize a Heart Attack? Quiz

    Science.gov (United States)

    ... Peripheral Artery Disease Venous Thromboembolism Aortic Aneurysm More Can You Recognize a Heart Attack? Updated:Sep 16, ... a Heart Attack Heart Attack Symptoms in Women “Can you recognize a heart attack?” Quiz • Understand Your ...

  3. Adsorption of trypsin on hydrophilic and hydrophobic surfaces

    NARCIS (Netherlands)

    Koutsopoulos, Sotirios; Patzsch, Katja; Bosker, Wouter T. E.; Norde, Willem

    2007-01-01

    The adsorption of trypsin onto polystyrene and silica surfaces was investigated by reflectometry, spectroscopic methods, and atomic force microscopy (AFM). The affinity of trypsin for the hydrophobic polystyrene surface was higher than that for the hydrophilic silica surface, but steady-state

  4. The trypsin-catalyzed hydrolysis of monomolecular films of lysylphosphatidylglycerol

    NARCIS (Netherlands)

    Gould, R.M.; Dawson, R.M.C.

    1972-01-01

    The hydrolysis by trypsin of the bacterial phospholipid, lysylphosphatidyl-glycerol has been studied at the air-water interface. High specific activity [14C]-lysylphosphatidylglycerol was prepared biosynthetically and the trypsin action followed by measuring the loss of surface radioactivity from a

  5. Potential Use of Atlantic Cod Trypsin in Biomedicine

    Science.gov (United States)

    Gudmundsdóttir, Ágústa; Hilmarsson, Hilmar; Stefansson, Bjarki

    2013-01-01

    Surface proteins of viruses and bacteria used for cell attachment and invasion are candidates for degradation by proteases. Trypsin from Atlantic cod (Gadus morhua) was previously demonstrated to have efficacy against influenza viruses in vitro and on skin. In this paper, cod trypsin is shown to be 3–12 times more effective in degrading large native proteins than its mesophilic analogue, bovine trypsin. This is in agreement with previous findings where cod trypsin was found to be the most active among twelve different proteases in cleaving various cytokines and pathological proteins. Furthermore, our results show that cod trypsin has high efficacy against herpes simplex virus type 1 (HSV-1) and the respiratory syncytial virus (RSV) in vitro. The results on the antipathogenic properties of cod trypsin are important because rhinovirus, RSV, and influenza are the most predominant pathogenic viruses in upper respiratory tract infections. Results from a clinical study presented in this paper show that a specific formulation containing cod trypsin was preferred for wound healing over other methods used in the study. Apparently, the high digestive ability of the cold-adapted cod trypsin towards large native proteins plays a role in its efficacy against pathogens and its positive effects on wounds. PMID:23555095

  6. Potential Use of Atlantic Cod Trypsin in Biomedicine

    Directory of Open Access Journals (Sweden)

    Ágústa Gudmundsdóttir

    2013-01-01

    Full Text Available Surface proteins of viruses and bacteria used for cell attachment and invasion are candidates for degradation by proteases. Trypsin from Atlantic cod (Gadus morhua was previously demonstrated to have efficacy against influenza viruses in vitro and on skin. In this paper, cod trypsin is shown to be 3–12 times more effective in degrading large native proteins than its mesophilic analogue, bovine trypsin. This is in agreement with previous findings where cod trypsin was found to be the most active among twelve different proteases in cleaving various cytokines and pathological proteins. Furthermore, our results show that cod trypsin has high efficacy against herpes simplex virus type 1 (HSV-1 and the respiratory syncytial virus (RSV in vitro. The results on the antipathogenic properties of cod trypsin are important because rhinovirus, RSV, and influenza are the most predominant pathogenic viruses in upper respiratory tract infections. Results from a clinical study presented in this paper show that a specific formulation containing cod trypsin was preferred for wound healing over other methods used in the study. Apparently, the high digestive ability of the cold-adapted cod trypsin towards large native proteins plays a role in its efficacy against pathogens and its positive effects on wounds.

  7. Trypsin Inhibitor Activity and Condensed Tannin Content in ...

    African Journals Online (AJOL)

    MICHAEL

    antinutrients trypsin inhibitor activity and condensed tannin content in nine landraces of bambara groundnut grown in three Southern African countries, ... nutrition is limited by antinutrients such as trypsin inhibitors and condensed tannins .... dry heat also need thorough investigations to determine if they are efficient in ...

  8. Interaction of gallic acid with trypsin analyzed by spectroscopy

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    Hao Song

    2015-06-01

    Full Text Available The interactions between trypsin and gallic acid (GA were investigated by means of fluorescence spectroscopy, UV-vis absorption spectroscopy, resonance light scattering (RLS spectroscopy, synchronous fluorescence spectroscopy, and enzymatic inhibition assay. It was found that GA can cause the fluorescence quenching of trypsin during the process of formation of GA-trypsin complex, resulting in inhibition of trypsin activity (IC50 = 3.9 × 10−6 mol/L. The fluorescence spectroscopic data showed that the quenching efficiency can reach about 80%. The binding constants were 1.9371 × 104 L/mol, 1.8192 × 104 L/mol, and 1.7465 × 104 L/mol at three temperatures, respectively. The thermodynamic parameters revealed that hydrogen bonds, van der Waals, hydrophobic, and electrostatic interactions were involved in the binding process of GA to trypsin. Molecular modeling studies illustrated a specific display of binding information and explained most of the experiment phenomena. The microenvironments of tryptophan and tyrosine residue in trypsin were changed by the GA. Results indicated that GA was a strong quencher and inhibitor of trypsin.

  9. Chemically modified, immobilized trypsin reactor with improved digestion efficiency

    NARCIS (Netherlands)

    Freije, J.R.; Mulder, P.P.; Werkman, W.; Rieux, L.; Niederlander, H.A G; Verpoorte, Sabeth; Bischoff, Rainer

    2005-01-01

    Tryptic digestion followed by identification using mass spectrometry is an important step in many proteomic studies. Here, we describe the preparation of immobilized, acetylated trypsin for enhanced digestion efficacy in integrated protein analysis platforms. Complete digestion of cytochrome c was

  10. Liver regeneration in trypsin-fed partially hepatectomized rats.

    Science.gov (United States)

    Gershbein, L L

    1993-01-01

    Young adult Sprague-Dawley rats were partially hepatectomized (two-thirds organ removal) and administered a basal diet supplemented with various animal- and plant-derived enzymes (trypsin, alpha-chymotrypsin, pepsin, lipase, alpha-amylase, malt diastase, ficin and bromelain) over a post-operative period of up to 10 days. Porcine or bovine dialyzed and lyophilized crystalline trypsin products containing 2400-3200 NF u/mg in addition to enteric-coated tablets with trypsin to chymotrypsin in a ratio of 6:1, were tested at supplementary levels of up to 4980 u/g ration. With the weight of tissue regenerated or the liver increment as indicator, trypsin in excess of 1000-1200 u/g ration proved inhibitory. This effect did not extend to alpha-chymotrypsin (levels of up to 4000 u/g diet) and the remaining 6 enzyme products specified above, nor to the s.c. injection of trypsin daily at 12,860 u/rat for the 1st 7 days. The last route promoted little change in increment with soy bean trypsin inhibitor (8.0 mg/rat daily for days 1 to 9). When a portion of the group fed a trypsin supplement of 2000 u/g was injected with phenobarbital i.p. at 80 mg/kg daily on each of the last 3 days, the resulting liver increment rose to the control range. As with lysine and arginine, acids of pertinence in tryptic proteolysis, no significant change was elicited by feeding a diet supplemented with peptone from tryptic digestion of casein. The enzyme-containing diets fed to sham-operated rats over a similar interval, did not affect the wet- or dry-liver weight per 100 g body weight. Microsomal parameters as total protein, cytochrome P-450 and the enzymes, aminopyrine demethylase and benzo[a]pyrene hydroxylase of livers from the partially hepatectomized or sham-operated rats fed trypsin and the other enzyme diets, presented no significant changes in the respective levels. The possible action of dietary trypsin in conjunction with inhibitors and growth factors controlling liver regeneration is

  11. Extraction purification and characterization of trypsin inhibitors from Andean seeds

    Directory of Open Access Journals (Sweden)

    Patricio Castillo

    2017-09-01

    Full Text Available This work established the conditions of covalent immobilization of trypsin on a Sepharose matrix, which could be applied for the purification of trypsin inhibitors. The higher values of retention of enzymatic activity and immobilized enzymatic activity were obtained with a Sepharose 6B-CL matrix, at room temperature, a pH value of 10.5, an enzymatic load of 25 mg/mL, and a minimum immobilization time of 12 hours, in order to obtain a stable immobilization. The most active trypsin inhibitors were selected through the comparison of, extracts obtained from the seeds of amaranth (Amaranthus caudatus L., pea (Pisum sativum, lupine or “chocho” (Lupinus mutabilis, bean (Phaseolus vulgaris and “sangorache” (Amaranthus hybridus L.. The inhibitors were partially purified using centrifugal ultrafiltration, heat treatment, and TCA precipitation. The permeated and retained fractions of “sangorache” were selected as the most active trypsin inhibitors, and they were selectively purified using affinity chromatography in a Trypsin - Glyoxyl - Sepharose 6B-CL matrix. The kinetic characterization showed the presence of two inhibitors; the first one corresponded to a competitive inhibitor, while the second one behaved as a mixed inhibitor.

  12. Gelatin-Enabled Microsensor for Pancreatic Trypsin Sensing

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    George Banis

    2018-01-01

    Full Text Available Digestive health is critically dependent on the secretion of enzymes from the exocrine pancreas to the duodenum via the pancreatic duct. Specifically, pancreatic trypsin is a major protease responsible for breaking down proteins for absorption in the small intestine. Gelatin-based hydrogels, deposited in the form of thin films, have been studied as potential sensor substrates that hydrolyze in the presence of trypsin. In this work, we (1 investigate gelatin as a sensing material; (2 develop a fabrication strategy for coating sensor surfaces; and (3 implement a miniaturized impedance platform for measuring activity levels of pancreatic trypsin. Using impedance spectroscopy, we evaluate gelatin’s specificity and rate of degradation when exposed to a combination of pancreatic enzymes in neutral solution representative of the macromolecular heterogeneity present in the duodenal environment. Our findings suggest gelatin’s preferential degradation to trypsin compared to enzymes such as lipase and amylase. We further observe their interference with trypsin behavior in equivalent concentrations, reducing film digestion by as much as 83% and 77%, respectively. We achieve film patterns in thicknesses ranging from 300–700 nm, which we coat over interdigitated finger electrode sensors. Finally, we test our sensors over several concentrations to emulate the range of pancreatic secretions. Ultimately, our microsensor will serve as the foundation for developing in situ sensors toward diagnosing pancreatic pathologies.

  13. Porphyrin Induced Laser Deactivation of Trypsinogen-Trypsin Conversion

    Science.gov (United States)

    Perido, Joanna; Brancaleon, Lorenzo

    2015-03-01

    Pancreatitis is caused by the inflammation of the pancreas, where the digestive enzyme trypsin is activated from the precursor enzyme trypsinogen while still in the pancreas. The presence of trypsin in the pancreas causes auto-activation of trypsinogen, resulting in greater inflammation and auto-digestion of the pancreas. In severe cases, this cascade effect can lead to organ failure, diabetes, and pancreatic cancer. Our hypothesis is that if trypsinogen is prevented from auto-activating into trypsin, then this cascade can be stopped. We propose to do this by inducing conformational changes in trypsinogen when bound to a photoactive porphyrin dye. Porphyrins are comprised of four linked heterocyclic groups forming a flat ring, and bind well with proteins such as trypsinogen. In this study we used spectroscopic techniques to probe the binding of meso-tetrakis (4-sulfonatephenyl) porphyrin (TSPP) to trypsinogen in vitro, as a preliminary step to then prompt and characterize conformational changes of trypsinogen through irradiation. If conformational changes are detected the trypsinogen will be tested for trypsin inactivation. This investigation may provide promising initial results to the possible use of porphyrins as an inhibitor of the self-activation of trypsinogen into trypsin, and a potential inhibitor of pancreatitis. MARC*U-STAR.

  14. Elucidation of different cold-adapted Atlantic cod (Gadus morhua) trypsin X isoenzymes.

    Science.gov (United States)

    Stefansson, Bjarki; Sandholt, Gunnar B; Gudmundsdottir, Ágústa

    2017-01-01

    Trypsins from Atlantic cod (Gadus morhua), consisting of several isoenzymes, are highly active cold-adapted serine proteases. These trypsins are isolated for biomedical use in an eco-friendly manner from underutilized seafood by-products. Our group has explored the biochemical properties of trypsins and their high potential in biomedicine. For broader utilization of cod trypsins, further characterization of biochemical properties of the individual cod trypsin isoenzymes is of importance. For that purpose, a benzamidine purified trypsin isolate from Atlantic cod was analyzed. Anion exchange chromatography revealed eight peaks containing proteins around 24kDa with tryptic activity. Based on mass spectrometric analysis, one isoenzyme gave the best match to cod trypsin I and six isoenzymes gave the best match to cod trypsin X. Amino terminal sequencing of two of these six trypsin isoenzymes showed identity to cod trypsin X. Three sequence variants of trypsin X were identified by cDNA analysis demonstrating that various forms of this enzyme exist. One trypsin X isoenzyme was selected for further characterization based on abundance and stability. Stepwise increase in catalytic efficiency (kcat/Km) of this trypsin X isoenzyme was obtained with substrates containing one to three amino acid residues. The study demonstrates that the catalytic efficiency of this trypsin X isoenzyme is comparable to that of cod trypsin I, the most abundant and highly active isoenzyme in the benzamidine cod trypsin isolate. Differences in pH stability and sensitivity to inhibitors of the trypsin X isoenzyme compared to cod trypsin I were detected that may be important for practical use. Copyright © 2016. Published by Elsevier B.V.

  15. FDA Recognized Consensus Standards

    Data.gov (United States)

    U.S. Department of Health & Human Services — This database consists of those national and international standards recognized by FDA which manufacturers can declare conformity to and is part of the information...

  16. Recognizing teen depression

    Science.gov (United States)

    ... medlineplus.gov/ency/patientinstructions/000648.htm Recognizing teen depression To use the sharing features on this page, ... life. Be Aware of the Risk for Teen Depression Your teen is more at risk for depression ...

  17. Cytogenetic study of Ascaris trypsin inhibitor in cultured human ...

    Indian Academy of Sciences (India)

    2009-04-01

    Apr 1, 2009 ... helminth (Souza et al. 2002; Oshiro et al. 2004). Previ- ously we reported the results on chromosome aberrations and sister chromatid changes in human lymphocytes exposed to. Ascaris trypsin inhibitor in the absence of metabolic S9 acti- vation (Blaszkowska et al. 2004). In these experimental con- ditions ...

  18. Page 1 Trypsinated nylon surfaces 8} immobilize onto various ...

    Indian Academy of Sciences (India)

    Trypsinated nylon surfaces 8} immobilize onto various polymer surfaces using suitable catalysts. Our above investigations, are only preliminary in nature, basically, to explore possibilities in this area which appears to be interesting for certain enzymes suitable towards blood compatibility. References. Andrade J D, Lee M B, ...

  19. Orthosteric and Allosteric Regulation in Trypsin-Like Peptidases

    DEFF Research Database (Denmark)

    Kromann-Tofting, Tobias

    peptides and Camelid derived antibody fragments, so-called nanobodies. Allosteric regulation of activity in trypsin-like serine peptidases is in general poorly understood, as the propagation of the allosteric signal from the ligand binding site to the active site in some cases is subtle and based...

  20. Efficiency of inactivation of trypsin inhibitory activity in some selected ...

    African Journals Online (AJOL)

    Dr. J. T. Ekanem

    2005-02-24

    Feb 24, 2005 ... toxicity is largely eliminated. For example, phytohaemagglutinins and trypsin inhibitors. (TIs) are heat labile and as such are readily destroyed by treatment of plant material with moist heat or autoclaving, dry heat being less effective. Levels of oxalates, tannin, nitrate/nitrite and various other antinutrients in.

  1. Magnetic nanoparticles coated with polyaniline to stabilize immobilized trypsin

    Science.gov (United States)

    Maciel, J. C.; D. Mercês, A. A.; Cabrera, M.; Shigeyosi, W. T.; de Souza, S. D.; Olzon-Dionysio, M.; Fabris, J. D.; Cardoso, C. A.; Neri, D. F. M.; C. Silva, M. P.; Carvalho, L. B.

    2016-12-01

    It is reported the synthesis of magnetic nanoparticles via the chemical co-precipitation of Fe 3+ ions and their preparation by coating them with polyaniline. The electronic micrograph analysis showed that the mean diameter for the nanoparticles is ˜15 nm. FTIR, powder X-ray diffraction and Mössbauer spectroscopy were used to understand the chemical, crystallographic and 57Fe hyperfine structures for the two samples. The nanoparticles, which exhibited magnetic behavior with relatively high spontaneous magnetization at room temperature, were identified as being mainly formed by maghemite ( γFe2O3). The coated magnetic nanoparticles (sample labeled "mPANI") presented a real ability to bind biological molecules such as trypsin, forming the magnetic enzyme derivative (sample "mPANIG-Trypsin"). The amount of protein and specific activity of the immobilized trypsin were found to be 13±5 μg of protein/mg of mPANI (49.3 % of immobilized protein) and 24.1±0.7 U/mg of immobilized protein, respectively. After 48 days of storage at 4 ∘C, the activity of the immobilized trypsin was found to be 89 % of its initial activity. This simple, fast and low-cost procedure was revealed to be a promising way to prepare mPANI nanoparticles if technological applications addressed to covalently link biomolecules are envisaged. This route yields chemically stable derivatives, which can be easily recovered from the reaction mixture with a magnetic field and recyclable reused.

  2. activity of enzyme trypsin immobilized onto macroporous poly(epoxy

    African Journals Online (AJOL)

    dell

    macropores and the unique sponge-like morphology that can permit unhindered diffusion of solutes of practically any size. (Lozinsky et al. ... sodium acetate, sodium phosphate (Na3PO4), sodium bicarbonate (NaHCO3) and sodium hydroxide (NaOH) were from Merck. (Darmstadt, Germany). Equipment. For trypsin activity ...

  3. Supercritical fluid assisted production of micrometric powders of the labile trypsin and chitosan/trypsin composite microparticles.

    Science.gov (United States)

    Shen, Yu-Bin; Guan, Yi-Xin; Yao, Shan-Jing

    2015-07-15

    Supercritical fluid assisted atomization introduced by a hydrodynamic cavitation mixer (SAA-HCM) was used to prepare micrometric particles of a labile protein, i.e., trypsin from aqueous solution without use of any organic solvents. The trypsin particles precipitated had various morphologies under different process conditions, with particle diameters ranging from 0.2 to 4 μm. FTIR, SDS-PAGE, CD and fluorescence spectra were performed to analyze the structural stability of the protein, and trypsin retained above 70% of the biological activity. Besides, chitosan was selected as the polymer carrier in an effort to prepare trypsin composite microparticles via SAA-HCM process. The influences of chitosan molecular weight, polymer/protein ratio and solution concentration on the particle morphology and size distribution were investigated in detail. Non-coalescing spherical composite microparticles with a narrow particle distribution (0.2-3 μm) could be obtained. The SAA-HCM prepared particles were amorphous as demonstrated by XRD and had a loading efficiency about 90%. The protein release profiles of the composite microparticles were evaluated using both the immersion condition and a Franz diffusion cell. Finally, the distribution of the protein within the particles was characterized through CLSM analysis of FITC-labeled trypsin-loaded chitosan microparticles. The SAA-HCM process is demonstrated to be a protein-friendly and promising technique for production of protein and polymer/protein composite particles formulations from aqueous solutions for drug delivery systems. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Interaction of methotrexate with trypsin analyzed by spectroscopic and molecular modeling methods

    Science.gov (United States)

    Wang, Yanqing; Zhang, Hongmei; Cao, Jian; Zhou, Qiuhua

    2013-11-01

    Trypsin is one of important digestive enzymes that have intimate correlation with human health and illness. In this work, the interaction of trypsin with methotrexate was investigated by spectroscopic and molecular modeling methods. The results revealed that methotrexate could interact with trypsin with about one binding site. Methotrexate molecule could enter into the primary substrate-binding pocket, resulting in inhibition of trypsin activity. Furthermore, the thermodynamic analysis implied that electrostatic force, hydrogen bonding, van der Waals and hydrophobic interactions were the main interactions for stabilizing the trypsin-methotrexate system, which agreed well with the results from the molecular modeling study.

  5. Trypsin-mediated enzymatic degradation of type II collagen in the human vitreous

    Science.gov (United States)

    van Deemter, Mariëlle; Kuijer, Roel; Harm Pas, Hendri; Jacoba van der Worp, Roelofje; Hooymans, Johanna Martina Maria

    2013-01-01

    Purpose Aging of the vitreous body can result in sight-threatening pathology. One aspect of vitreous aging is liquefaction, which results from the vanishing of collagen fibrils. We investigated the possibility that trypsins are involved in vitreous type II collagen degradation. Methods Immunohistochemistry and western blotting were used for detecting and locating trypsin isoforms in the vitreous and retina of human donor eyes. The capability of the retina to produce these trypsins was analyzed with polymerase chain reaction. Whether the different trypsins degraded type II collagen was tested in vitro. The sizes of the in vitro induced type II collagen degradation products were compared to those present in the vitreous of human eyes of different ages. Results Trypsin-1 and trypsin-2 were detected in the vitreous. In the retina, messenger ribonucleic acid (mRNA) coding for trypsin-2, -3, and -4 was present. Using immunohistochemistry, trypsin-2 was detected in microglial cells located in the vitreous and the retina. All trypsin isoforms degraded type II collagen and produced degradation products of similar sizes as those present in the vitreous. Conclusions Trypsin-1 and trypsin-2 appear to have a function in the degradation of vitreous type II collagen. They could therefore have a role in the development of vitreous liquefaction. PMID:23882137

  6. Effects of Trypsinization on Viability of Equine Chondrocytes in Cell Culture

    Directory of Open Access Journals (Sweden)

    B. C. Sutradhar, J. Park, G. Hong, S. H. Choi and G. Kim*

    2010-10-01

    Full Text Available Trypsin is an essential reagent for routine cell culture work. In the cultivation of mammalian cells, it has been extensively used for cell isolation from tissues or cell dislodging in subculturing. It may damage the cell membrane in contact of cells during long trypsinization. However, there is no specific report on time-dependent effect of trypsinization on cells. In the present study, we investigated the time dependent effects of trypsinization on equine chondrocytes. Cell viability after trypsinization with 0.25% trypsin-EDTA for 5 to 60 minutes was quantified by trypan blue exclusion assay, propidium iodide-Hoechst double staining, flow cytometry analysis and XTT assay. The results showed that trypsin-EDTA decreased the proliferation of equine chondrocytes depending on the exposure time of trypsinization. After 20 and 60 minutes of trypsinization, the cell membranes were strongly affected and the percentages of viable cells reduced to 91% and 85% respectively detected by trypan blue exclusion assay. Similar results were observed both in flow cytometric evaluation and propidium iodide-Hoechst double staining. The XTT assay result also showed decreased cell viability with the extended time of trypsinization. In order to minimize the time dependant cytotoxicity of trypsinization, as minimum as short time exposure is suggestive that maximizes live cell isolation from tissue as well as subculture of equine chondrocytes or other cells.

  7. Comparative study of the binding of trypsin to caffeine and theophylline by spectrofluorimetry

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ruiyong, E-mail: wangry@zzu.edu.cn [Department of Chemistry, Zhengzhou University, Zhengzhou 450001 (China); Kang, Xiaohui [Department of Chemistry, Zhengzhou University, Zhengzhou 450001 (China); Wang, Ruiqiang [The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052 (China); Wang, Rui; Dou, Huanjing; Wu, Jing; Song, Chuanjun [Department of Chemistry, Zhengzhou University, Zhengzhou 450001 (China); Chang, Junbiao, E-mail: changjunbiao@zzu.edu.cn [Department of Chemistry, Zhengzhou University, Zhengzhou 450001 (China)

    2013-06-15

    The interactions between trypsin and caffeine/theophylline were investigated by fluorescence spectroscopy, UV–visible absorption spectroscopy, resonance light scattering and synchronous fluorescence spectroscopy under mimic physiological conditions. The results revealed that the fluorescence quenching of trypsin by caffeine and theophylline was the result of the formed complex of caffeine–trypsin and theophylline–trypsin. The binding constants and thermodynamic parameters at three different temperatures were obtained. The hydrophobic interaction was the predominant intermolecular forces to stabilize the complex. Results showed that caffeine was the stronger quencher and bound to trypsin with higher affinity than theophylline. -- Highlights: ► The fluorescence of trypsin can be quenched by caffeine or theophylline via hydrophobic contacts. ► Caffeine binds to trypsin with higher affinity than theophylline. ► The influence of molecular structure on the binding aspects is reported.

  8. Structure and Function of Trypsin-Loaded Fibrinolytic Liposomes

    OpenAIRE

    Anna Tanka-Salamon; Attila Bóta; András Wacha; Judith Mihály; Miklós Lovas; Krasimir Kolev

    2017-01-01

    Protease encapsulation and its targeted release in thrombi may contribute to the reduction of haemorrhagic complications of thrombolysis. We aimed to prepare sterically stabilized trypsin-loaded liposomes (SSLT) and characterize their structure and fibrinolytic efficiency. Hydrogenated soybean phosphatidylcholine-based SSLT were prepared and their structure was studied by transmission electron microscopy combined with freeze fracture (FF-TEM), Fourier transform infrared spectroscopy (FT-IR), ...

  9. Torque teno sus virus (TTSuV in cell cultures and trypsin.

    Directory of Open Access Journals (Sweden)

    Thais Fumaco Teixeira

    Full Text Available Torque teno sus virus (TTSuV, a member of the family Anelloviridae, is a single-stranded, circular DNA virus, widely distributed in swine populations. Presently, two TTSuV genogroups are recognized: Torque teno sus virus 1 (TTSuV1 and Torque teno sus virus 2 (TTSuV2. TTSuV genomes have been found in commercial vaccines for swine, enzyme preparations and other drugs containing components of porcine origin. However, no studies have been made looking for TTSuV in cell cultures. In the present study, a search for TTSuV genomes was carried out in cell culture lineages, in sera used as supplement for cell culture media as well as in trypsin used for cell disaggregation. DNA obtained from twenty-five cell lineages (ten from cultures in routine multiplication and fifteen from frozen ampoules, nine samples of sera used in cell culture media and five batches of trypsin were examined for the presence of TTSuV DNA. Fifteen cell lineages, originated from thirteen different species contained amplifiable TTSuV genomes, including an ampoule with a cell lineage frozen in 1985. Three cell lineages of swine origin were co-infected with both TTSuV1 and TTSuV2. One batch of trypsin contained two distinct TTSuV1 plus one TTSuV2 genome, suggesting that this might have been the source of contamination, as supported by phylogenetic analyses of sequenced amplicons. Samples of fetal bovine and calf sera used in cell culture media did not contain amplifiable TTSuV DNA. This is the first report on the presence of TTSuV as contaminants in cell lineages. In addition, detection of the viral genome in an ampoule frozen in 1985 provides evidence that TTSuV contamination is not a recent event. These findings highlight the risks of TTSuV contamination in cell cultures, what may be source for contamination of biological products or compromise results of studies involving in vitro multiplied cells.

  10. Interaction between 8-methoxypsoralen and trypsin: Monitoring by spectroscopic, chemometrics and molecular docking approaches

    Science.gov (United States)

    Liu, Yingying; Zhang, Guowen; Zeng, Ni; Hu, Song

    2017-02-01

    8-Methoxypsoralen (8-MOP) is a naturally occurring furanocoumarin with various biological activities. However, there is little information on the binding mechanism of 8-MOP with trypsin. Here, the interaction between 8-MOP and trypsin in vitro was determined by multi-spectroscopic methods combined with the multivariate curve resolution-alternating least squares (MCR-ALS) chemometrics approach. An expanded UV-vis spectral data matrix was analysed by MCR-ALS, the concentration profiles and pure spectra for the three reaction species (trypsin, 8-MOP and 8-MOP-trypsin) were obtained to monitor the interaction between 8-MOP and trypsin. The fluorescence data suggested that a static type of quenching mechanism occurred in the binding of 8-MOP to trypsin. Hydrophobic interaction dominated the formation of the 8-MOP-trypsin complex on account of the positive enthalpy and entropy changes, and trypsin had one high affinity binding site for 8-MOP with a binding constant of 3.81 × 104 L mol- 1 at 298 K. Analysis of three dimensional fluorescence, UV-vis absorption and circular dichroism spectra indicated that the addition of 8-MOP induced the rearrangement of the polypeptides carbonyl hydrogen-bonding network and the conformational changes in trypsin. The molecular docking predicted that 8-MOP interacted with the catalytic residues His57, Asp102 and Ser195 in trypsin. The binding patterns and trypsin conformational changes may result in the inhibition of trypsin activity. This study has provided insights into the binding mechanism of 8-MOP with trypsin.

  11. Magnetic nanoparticles coated with polyaniline to stabilize immobilized trypsin

    Energy Technology Data Exchange (ETDEWEB)

    Maciel, J. C., E-mail: jackeline-maciel@hotmail.com [Universidade Federal de Roraima (Brazil); Mercês, A. A. D.; Cabrera, M. [Universidade Federal de Pernambuco, Laboratório de Imunopatologia Keizo Asami (Brazil); Shigeyosi, W. T. [Universidade Federal de São Carlos, Departamento de Física (Brazil); Souza, S. D. de; Olzon-Dionysio, M.; Fabris, J. D. [Universidade Federal dos Vales de Jequitinhonha e Mucuri (Brazil); Cardoso, C. A. [Universidade Federal de São Carlos, Departamento de Física (Brazil); Neri, D. F. M. [Universidade Federal do Vale do São Francisco (Brazil); Silva, M. P. C.; Carvalho, L. B. [Universidade Federal de Pernambuco, Laboratório de Imunopatologia Keizo Asami (Brazil)

    2016-12-15

    It is reported the synthesis of magnetic nanoparticles via the chemical co-precipitation of Fe {sup 3+} ions and their preparation by coating them with polyaniline. The electronic micrograph analysis showed that the mean diameter for the nanoparticles is ∼15 nm. FTIR, powder X-ray diffraction and Mössbauer spectroscopy were used to understand the chemical, crystallographic and {sup 57}Fe hyperfine structures for the two samples. The nanoparticles, which exhibited magnetic behavior with relatively high spontaneous magnetization at room temperature, were identified as being mainly formed by maghemite (γFe{sub 2}O{sub 3}). The coated magnetic nanoparticles (sample labeled “mPANI”) presented a real ability to bind biological molecules such as trypsin, forming the magnetic enzyme derivative (sample “mPANIG-Trypsin”). The amount of protein and specific activity of the immobilized trypsin were found to be 13±5 μg of protein/mg of mPANI (49.3 % of immobilized protein) and 24.1±0.7 U/mg of immobilized protein, respectively. After 48 days of storage at 4 {sup ∘}C, the activity of the immobilized trypsin was found to be 89 % of its initial activity. This simple, fast and low-cost procedure was revealed to be a promising way to prepare mPANI nanoparticles if technological applications addressed to covalently link biomolecules are envisaged. This route yields chemically stable derivatives, which can be easily recovered from the reaction mixture with a magnetic field and recyclable reused.

  12. Human and bovine spinal disc mechanics subsequent to trypsin injection

    Directory of Open Access Journals (Sweden)

    Jeremy Alsup

    2017-10-01

    The Translational Potential of this Article: Preclinical testing of novel spinal devices is essential to the design validation and regulatory processes, but current testing techniques rely on cadaveric testing of primarily older spines with essentially random amounts of disc degeneration. The present work investigates the viability of using trypsin injections to create a more uniform preclinical model of disc degeneration from a mechanics perspective, for the purpose of testing spinal devices. Such a model would facilitate translation of new spinal technologies to clinical practice.

  13. Chitosan nanoencapsulated exogenous trypsin biomimics zymogen-like enzyme in fish gastrointestinal tract.

    Directory of Open Access Journals (Sweden)

    Rakhi Kumari

    Full Text Available Exogenous proteolytic enzyme supplementation is required in certain disease conditions in humans and animals and due to compelling reasons on use of more plant protein ingredients and profitability in animal feed industry. However, limitations on their utility in diet are imposed by their pH specificity, thermolabile nature, inhibition due to a variety of factors and the possibility of intestinal damage. For enhancing the efficacy and safety of exogenous trypsin, an efficient chitosan (0.04% nanoencapsulation-based controlled delivery system was developed. An experiment was conducted for 45 days to evaluate nanoencapsulated trypsin (0.01% and 0.02% along with 0.02% bare trypsin and 0.4% chitosan nanoparticles against a control diet on productive efficiency (growth rate, feed conversion and protein efficiency ratio, organo-somatic indices, nutrient digestibility, tissue enzyme activities, hematic parameters and intestinal histology of the fish Labeo rohita. All the synthesized nanoparticles were of desired characteristics. Enhanced fish productive efficiency using nanoencapsulated trypsin over its bare form was noticed, which corresponded with enhanced (P<0.01 nutrient digestibility, activity of intestinal protease, liver and muscle tissue transaminases (alanine and aspartate and dehydrogenases (lactate and malate, serum blood urea nitrogen and serum protein profile. Intestinal tissues of fish fed with 0.02% bare trypsin showed broadened, marked foamy cells with lipid vacuoles. However, villi were healthier in appearance with improved morphological features in fish fed with nanoencapsulated trypsin than with bare trypsin, and the villi were longer in fish fed with 0.01% nanoencapsulated trypsin than with 0.02% nanoencapsulated trypsin. The result of this premier experiment shows that nanoencapsulated trypsin mimics zymogen-like proteolytic activity via controlled release, and hence the use of 0.01% nanoencapsulated trypsin (in chitosan

  14. [Effect of the Hydrogel Carrier Structure on the Activity of Immobilized Trypsin].

    Science.gov (United States)

    Valuev, L I; Valuev, I L; Vanchugova, L V; Valueva, T A

    2015-01-01

    The dependence of the activity of trypsin immobilized in polyacrylamide hydrogel on the hydrogel swelling ratio, the size distribution of its pores, and the means of enzyme binding has been studied. It has been shown that the most favorable conditions for immobilized trypsin are provided upon its binding to hydrogel via trypsin macromonomer copolymerization with acrylamide and a linking agent in the presence of a modifier that limits polymer chain growth.

  15. Effects of bisphenol S on the structures and activities of trypsin and pepsin.

    Science.gov (United States)

    Wang, Yan-Qing; Zhang, Hong-Mei

    2014-11-19

    The effects of bisphenol S on the structures and activities of trypsin and pepsin were investigated by various methods like UV-visible absorbance, fluorescence, circular dichroism, and molecular docking. The secondary and tertiary structures of trypsin and pepsin were altered by bisphenol S binding, which resulted in the loosening of the skeletons of trypsin and pepsin. In addition, bisphenol S induced microenvironmental changes around tyrosine and tryptophan residues of trypsin and pepsin. The activity experimental results showed that the activity of pepsin decreases obviously with the increasing concentration of BPS, while the activity of trypsin does not change remarkably. The binding and thermodynamic parameters obtained by molecular docking and fluorescence spectroscopy showed that the bindings of bisphenol S to trypsin and pepsin were spontaneous processes and hydrogen bonding and hydrophobic interactions played a vital role in stabilizing the bisphenol S-trypsin and bisphenol S-pepsin complexes. The binding constants (K(A)) of bisphenol S with trypsin were 7.42 × 10(4) (298 K) and 5.91 × 10(4) L/mol (310 K), and those of pepsin were 5.78 × 10(4) (298 K) and 4.44 × 10(4) L/mol (310 K). Moreover, there was one main kind of binding site for bisphenol S on trypsin or pepsin.

  16. Digestive enzyme activity and mRNA level of trypsin in embryonic redclaw crayfish, Cherax quadricarnatus

    Science.gov (United States)

    Luo, Wen; Zhao, Yunlong; Zhou, Zhongliang; An, Chuanguang; Ma, Qiang

    2008-02-01

    The digestive enzyme activity and mRNA level of trypsin during the embryonic development of Cherax quadricarinatus were analyzed using biochemical and Fluorogenic Quantitative PCR (FQ—PCR) methods. The results show that the activities of trypsin and chymotrypsin had two different change patterns. Trypsin specific activity increased rapidly in the early stages of development and still remained high in preparation for the hatch stage. However, chymotrypsin activity peaked in stage 4 of embryonic development and decreased significantly in the last stage. The mRNA level of trypsin was elevated in all stages and two peak values were observed in stages 2 and 5 respectively. The results indicate that trypsin is very important for the utilization of the yolk during embryonic development and for the assimilation of dietary protein for larvae. The gene of trypsin is probably regulated at transcriptional level. The mRNA levels of trypsin can reflect not only trypsin activity, but also the regulatory mechanism for expression of trypsin gene to a certain degree.

  17. Structure and Function of Trypsin-Loaded Fibrinolytic Liposomes

    Directory of Open Access Journals (Sweden)

    Anna Tanka-Salamon

    2017-01-01

    Full Text Available Protease encapsulation and its targeted release in thrombi may contribute to the reduction of haemorrhagic complications of thrombolysis. We aimed to prepare sterically stabilized trypsin-loaded liposomes (SSLT and characterize their structure and fibrinolytic efficiency. Hydrogenated soybean phosphatidylcholine-based SSLT were prepared and their structure was studied by transmission electron microscopy combined with freeze fracture (FF-TEM, Fourier transform infrared spectroscopy (FT-IR, and small-angle X-ray scattering (SAXS. Fibrinolytic activity was examined at 45, 37, or 24°C on fibrin or plasma clots with turbidimetric and permeation-driven lysis assays. Trypsin was shown to be attached to the inner surface of vesicles (SAXS and FF-TEM close to the lipid hydrophilic/hydrophobic interface (FT-IR. The thermosensitivity of SSLT was evidenced by enhanced fibrinolysis at 45°C: time to reduce the maximal turbidity to 20% decreased by 8.6% compared to 37°C and fibrin degradation product concentration in the permeation lysis assay was 2-fold to 5-fold higher than that at 24°C. SSLT exerted its fibrinolytic action on fibrin clots under both static and dynamic conditions, whereas plasma clot dissolution was observed only in the permeation-driven assay. The improved fibrinolytic efficiency of SSLT under dynamic conditions suggests that they may serve as a novel therapeutic candidate for dissolution of intravascular thrombi, which are typically exposed to permeation forces.

  18. Recognizing and Treating Eye Injuries

    Science.gov (United States)

    ... Patient Stories Español Eye Health / Tips & Prevention Eye Injuries Sections Preventing Eye Injuries Recognizing and Treating Eye ... to Safer Champagne Celebrations Recognizing and Treating Eye Injuries Leer en Español: Reconociendo las Lesiones de los ...

  19. The Duty to Recognize Culture

    DEFF Research Database (Denmark)

    Nielsen, Morten Ebbe Juul

    2012-01-01

    On Taylor and Honneth's theories of recognition and whether one can derive a "duty to recognize Culture" from these......On Taylor and Honneth's theories of recognition and whether one can derive a "duty to recognize Culture" from these...

  20. Purification and characterization of trypsin produced by gut bacteria from Anticarsia gemmatalis.

    Science.gov (United States)

    Pilon, Franciny Martins; Silva, Camila da Rocha; Visôtto, Liliane Evangelista; Barros, Rafael de Almeida; da Silva Júnior, Neilier Rodrigues; Campos, Wellington Garcia; de Almeida Oliveira, Maria Goreti

    2017-10-01

    Purification of active trypsin in the digestive process of insects is essential for the development of potent protease inhibitors (PIs) as an emerging pest control technology and research into insect adaptations to dietary PIs. An important aspect is the presence of proteolytic microorganisms, which contribute to host nutrition. Here, we purified trypsins produced by bacteria Bacillus cereus, Enterococcus mundtii, Enterococcus gallinarum, and Staphylococcus xylosus isolated from the midgut of Anticarsia gemmatalis. The trypsins had a molecular mass of approximately 25 kDa. The enzymes showed increased activity at 40°C, and they were active at pH values 7.5-10. Aprotinin, bis-benzamidine, and soybean Kunitz inhibitor (SKTI) significantly inhibited trypsin activity. The l-1-tosyl-amido-2-phenylethylchloromethyl ketone (TPCK), pepstatin A, E-64, ethylenediamine tetraacetic acid, and calcium ions did not affect the enzyme activity at the concentrations tested. We infer the purified trypsins do not require calcium ions, by which they differ from the trypsins of other microorganisms and the soluble and insoluble trypsins characterized from A. gemmatalis. These data suggest the existence of different isoforms of trypsin in the velvetbean caterpillar midguts. © 2017 Wiley Periodicals, Inc.

  1. A spectroscopic and thermal stability study on the interaction between putrescine and bovine trypsin.

    Science.gov (United States)

    Momeni, Lida; Shareghi, Behzad; Saboury, Ali A; Farhadian, Sadegh; Reisi, Fateme

    2017-01-01

    The interaction of putrescine with bovine trypsin was investigated using steady state thermal stability, intrinsic fluorescence, UV-vis spectroscopy, far and near- UV circular dichroism and kinetic techniques, as well as molecular docking. The Stern-Volmer quenching constants for the trypsin- putrescine complex were calculated revealing that putrescine interacted with trypsin via the static fluorescence quenching. The enthalpy and entropy change values and the molecular docking technique revealed that hydrogen bonds and van der Waals forces play a major role in the binding process. Upon putrescine conjugation, the V max value and the k cat /K m values of the enzyme was increased. The results of UV absorbance, circular dichroism and fluorescence techniques demonstrated that the micro environmental changes in trypsin were induced by the binding of putrescine, leading to changes in its secondary structure. The thermal stability of trypsin- putrescine complex was enhanced more significantly, as compared to that of the native trypsin. The increased thermal stability of trypsin- putrescine complex might be due to the lower surface hydrophobicity and the higher hydrogen bond formation after putrescine modification, as reflected in the increase of UV absorbance and the quenching of fluorescence spectra. It was concluded that the binding of putrescine changed trypsin structure and function. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Synthesis and Characterization of Highly Stabilized Polymer–Trypsin Conjugates with Autolysis Resistance

    Directory of Open Access Journals (Sweden)

    Yasushi Sasai

    2016-12-01

    Full Text Available Protein digestion by trypsin has been widely used in many industrial and research applications. However, extensive use of trypsin is limited because of the rapid decrease in enzymatic activity caused by autolysis at optimal pH and temperature. To improve the enzymatic performance of trypsin, we synthesized highly stabilized polymer–trypsin conjugates using vinylmethylether-maleic acid copolymer (VEMAC via multi-point attachment. The VEMAC modification significantly enhanced the thermal stability of trypsin, and the resulting conjugates showed a strong resistance to autolysis. VEMAC-modified trypsin (VEMAC-Tryp showed maximum activity at 55 °C and at 1.4-fold higher levels than that of unmodified trypsin. Bovine serum albumin was effectively digested by VEMAC-Tryp, indicating that the modified trypsin can be used for digestion of high molecular weight substrates. VEMAC modification is a simple and cost-effective strategy to obtain fully active modified enzymes, and may be used to develop bioreactors.

  3. FRET-based modified graphene quantum dots for direct trypsin quantification in urine

    Energy Technology Data Exchange (ETDEWEB)

    Poon, Chung-Yan; Li, Qinghua [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region (Hong Kong); Zhang, Jiali; Li, Zhongping [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region (Hong Kong); Research Center of Environmental Science and Engineering, School of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006 (China); Dong, Chuan [Research Center of Environmental Science and Engineering, School of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006 (China); Lee, Albert Wai-Ming; Chan, Wing-Hong [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region (Hong Kong); Li, Hung-Wing, E-mail: hwli@hkbu.edu.hk [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region (Hong Kong)

    2016-04-21

    A versatile nanoprobe was developed for trypsin quantification with fluorescence resonance energy transfer (FRET). Here, fluorescence graphene quantum dot is utilized as a donor while a well-designed coumarin derivative, CMR2, as an acceptor. Moreover, bovine serum albumin (BSA), as a protein model, is not only served as a linker for the FRET pair, but also a fluorescence enhancer of the quantum dots and CMR2. In the presence of trypsin, the FRET system would be destroyed when the BSA is digested by trypsin. Thus, the emission peak of the donor is regenerated and the ratio of emission peak of donor/emission peak of acceptor increased. By the ratiometric measurement of these two emission peaks, trypsin content could be determined. The detection limit of trypsin was found to be 0.7 μg/mL, which is 0.008-fold of the average trypsin level in acute pancreatitis patient's urine suggesting a high potential for fast and low cost clinical screening. - Highlights: • A FRET-based biosensor was developed for direct quantification of trypsin. • Fast and sensitive screening of pancreatic disease was facilitated. • The direct quantification of trypsin in urine samples was demonstrated.

  4. Carbohydrate as covalent crosslink in human inter-alpha-trypsin inhibitor

    DEFF Research Database (Denmark)

    Jessen, T E; Faarvang, K L; Ploug, M

    1988-01-01

    The primary structure of inter-alpha-trypsin inhibitor is partially elucidated, but controversy about the construction of the polypeptide backbone still exists. We present evidence suggesting that inter-alpha-trypsin inhibitor represents a novel plasma protein structure with two separate polypept...... polypeptide chains covalently crosslinked only by carbohydrate (chondroitin sulphate)....

  5. Trypsin isozymes in the lobster Panulirus argus (Latreille, 1804): from molecules to physiology.

    Science.gov (United States)

    Perera, Erick; Rodríguez-Viera, Leandro; Perdomo-Morales, Rolando; Montero-Alejo, Vivian; Moyano, Francisco Javier; Martínez-Rodríguez, Gonzalo; Mancera, Juan Miguel

    2015-01-01

    Trypsin enzymes have been studied in a wide variety of animal taxa due to their central role in protein digestion as well as in other important physiological and biotechnological processes. Crustacean trypsins exhibit a high number of isoforms. However, while differences in properties of isoenzymes are known to play important roles in regulating different physiological processes, there is little information on this aspect for decapod trypsins. The aim of this review is to integrate recent findings at the molecular level on trypsin enzymes of the spiny lobster Panulirus argus, into higher levels of organization (biochemical, organism) and to interpret those findings in relation to the feeding ecology of these crustaceans. Trypsin in lobster is a polymorphic enzyme, showing isoforms that differ in their biochemical features and catalytic efficiencies. Molecular studies suggest that polymorphism in lobster trypsins may be non-neutral. Trypsin isoenzymes are differentially regulated by dietary proteins, and it seems that some isoenzymes have undergone adaptive evolution coupled with a divergence in expression rate to increase fitness. This review highlights important but poorly studied issues in crustaceans in general, such as the relation among trypsin polymorphism, phenotypic (digestive) flexibility, digestion efficiency, and feeding ecology.

  6. Kunitz-type trypsin inhibitor with high stability from Spinacia oleracea L. seeds.

    Science.gov (United States)

    Kang, Zhuang; Jiang, Jia-hong; Wang, Dong; Liu, Ke; Du, Lin-fang

    2009-01-01

    The trypsin inhibitor SOTI was isolated from Spinacia oleracea L. seeds through ammonium sulfate precipitation, Sepharose 4B-trypsin affinity chromatography, and Sephadex G-75 chromatography. This typical Kunitz inhibitor showed remarkable stability to heat, pH, and denaturant. It retained 80% of its activity against trypsin after boiling for 20 min, and more than 90% activity when treated with 6 M guanidine hydrochloride. The formation of stable SOTI-trypsin complex (K(i) = 2.3x10(-6) M) is consistent with significant inhibitory activity of SOTI against trypsin-like proteinases present in the larval midgut of Pieris rapae. Sequences of SOTI fragments showed homology with other inhibitors.

  7. Trypsinogen-trypsin transition: a molecular dynamics study of induced conformational change in the activation domain.

    Science.gov (United States)

    Brünger, A T; Huber, R; Karplus, M

    1987-08-11

    The trypsinogen to trypsin transition has been investigated by a stochastic boundary molecular dynamics simulation that included a major portion of the trypsin molecule and the surrounding solvent. Attention focused on the "activation domain", which crystallographic studies have shown to be ordered in trypsin and disordered in its zymogen, trypsinogen. The chain segments that form the activation domain were found to exhibit large fluctuations during the simulation of trypsin. To model a difference between trypsin and trypsinogen, the N-terminal residues Ile-16 and Val-17 were removed in the former and replaced by water molecules. As a result of the perturbation, a structural drift of 1-2 A occurred that is limited to the activation domain. Glycine residues are found to act as hinges for the displaced chain segments.

  8. [Chymotrypsin and trypsin inhibitor isolated from potato tubers].

    Science.gov (United States)

    Revina, T A; Parfenov, I A; Gvozdeva, E L; Gerasimova, N G; Valueva, T A

    2011-01-01

    Potato Kunitz-type chymotrypsin inhibitor (PKCI-23) was isolated from potato tubers (Solanum tuberosum L., Zhukov's Jubilee breed) and purified to a homogenous state. The protein was purified by gel-filtration chromatography and ion-exchange chromatography using Sephadex G-75 and CM-Sepharose CL-6B, respectively. PKCI-23 protein has been shown to inhibit both chymotrypsin and trypsin with equal efficacy, forming equimolar complexes with these enzymes. However, much weaker inhibitory effect of PKCI-23 has been observed for Carlsberg subtilisin. The N-terminal 20 amino acid sequence of PKCI-23 has been sequenced. PKCI-23 has been shown to suppress, with different efficacy, the growth and development of pathogenic microorganisms Fusarium culmorum (Wm. G. Sm.) Sacc. and Phytophtora infestans (Mont.) de Bary that infect potato.

  9. Recognizing child maltreatment in Bangladesh.

    Science.gov (United States)

    Khan, N Z; Lynch, M A

    1997-08-01

    Concern is increasing in Bangladesh over child abuse, neglect, and exploitation. Children from all walks of life are being treated at the Child Development Center (CDC) Dhaka Shishu Hospital for neurodevelopmental problems resulting from abuse and neglect. Efforts to protect children from sexual harassment result in girls being isolated at home or married at an early age. Some young brides are eventually abandoned and forced into prostitution. Early marriage reflects the lack of acknowledgement of a period of adolescence and the belief that puberty is a marker of adulthood. Many girls aged 8-16 are employed as live-in domestic servants, and many suffer sexual as well as emotional abuse. Garment factories, on the other hand, offer girls an escape from extreme poverty, domestic service, and early marriage but are threatened by forces that condemn child labor. Rather than ending such opportunities, employers should be encouraged to provide employees with educational and welfare facilities. The CDC seeks to explore the extent and depth of the problem of child abuse while recognizing the special circumstances at work in Bangladesh. It is also necessary to raise awareness of these issues and of the discrepancies between the law and cultural practices. For example, the legal marriage age of 18 years for a woman and 21 years for a man is often ignored. Additional forms of abuse receiving the attention of women's organizations and human rights groups include the trafficking of children. A network of concerned organizations should be created to work against the child abuse, neglect, and exploitation that Bangladesh has pledged to overcome by signing the UN Convention on the Rights of the Child.

  10. Production of β-Lactoglobulin hydrolysates by monolith based immobilized trypsin reactors.

    Science.gov (United States)

    Mao, Yuhong; Černigoj, Urh; Zalokar, Viktor; Štrancar, Aleš; Kulozik, Ulrich

    2017-11-01

    Tryptic hydrolysis of β-Lactoglobulin (β-Lg) is attracting more and more attention due to the reduced allergenicity and the functionality of resulting hydrolysates. To produce hydrolysates in an economically viable way, immobilized trypsin reactors (IMTRs), based on polymethacrylate monolith with pore size 2.1 μm (N1) and 6 μm (N2), were developed and used in a flow-through system. IMTRs were characterized in terms of permeability and enzymatic activity during extensive usage. N1 showed twice the activity compared with N2, correlating well with its almost two times higher amount of immobilized trypsin. N2 showed high stability over 18 cycles, as well as over more than 30 weeks during storage. The efficiency of IMTRs on hydrolyzing β-Lg was compared with free trypsin, and the resulting hydrolysates were analyzed by MALDI-TOF/MS. The final hydrolysis degree by N1 reached 9.68% (86.58% cleavage sites) within 4 h, while only around 6% (53.67% cleavage sites) by 1.5 mg of free trypsin. Peptides analysis showed the different preference between immobilized trypsin and free trypsin. Under the experimental conditions used in this study, the potential cleavage site Lys135 -Phe136 was resistant against the immobilized trypsin in N1. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Immobilization of Trypsin in Lignocellulosic Waste Material to Produce Peptides with Bioactive Potential from Whey Protein

    Directory of Open Access Journals (Sweden)

    Juliana Cristina Bassan

    2016-05-01

    Full Text Available In this study, trypsin (Enzyme Comission 3.4.21.4 was immobilized in a low cost, lignocellulosic support (corn cob powder—CCP with the goal of obtaining peptides with bioactive potential from cheese whey. The pretreated support was activated with glyoxyl groups, glutaraldehyde and IDA-glyoxyl. The immobilization yields of the derivatives were higher than 83%, and the retention of catalytic activity was higher than 74%. The trypsin-glyoxyl-CCP derivative was thermally stable at 65 °C, a value that was 1090-fold higher than that obtained with the free enzyme. The trypsin-IDA-glyoxyl-CCP and trypsin-glutaraldehyde-CCP derivatives had thermal stabilities that were 883- and five-fold higher, respectively, then those obtained with the free enzyme. In the batch experiments, trypsin-IDA-glyoxyl-CCP retained 91% of its activity and had a degree of hydrolysis of 12.49%, while the values for trypsin-glyoxyl-CCP were 87% and 15.46%, respectively. The stabilized derivative trypsin-glyoxyl-CCP was also tested in an upflow packed-bed reactor. The hydrodynamic characterization of this reactor was a plug flow pattern, and the kinetics of this system provided a relative activity of 3.04 ± 0.01 U·g−1 and an average degree of hydrolysis of 23%, which were suitable for the production of potentially bioactive peptides.

  12. Application of Frontal Affinity Chromatography to Study the Biomolecular Interactions with Trypsin.

    Science.gov (United States)

    Hu, YuanYuan; Qian, Junqing; Guo, Hui; Jiang, ShengLan; Zhang, Zheng

    2015-07-01

    Trypsin is a serine protease that has been proposed as a potential therapeutic target for metabolic disorders and malignancy diseases, thus the identification of biomolecular interactions of compounds to trypsin could be of great therapeutic importance. In this study, trypsin was immobilized on a monolithic silica capillary column via sol-gel. The binding properties of four small molecules (daidzin, genistin, matrine and oxymatrine) to trypsin were examined using the trypsin affinity columns by frontal analysis. The results indicate that the matrine (dissociation constant, Kd = 7.904 μM) has stronger interaction with trypsin than the oxymatrine (Kd = 8.204 μM), whereas daidzin and genistin were nearly have no affinity with trypsin. The results demonstrated that the frontal affinity chromatography can be used for the direct determination of protein-protease inhibitor binding interactions and have several significant advantages, including easy fabricating, reproducible, minimal technological requirements and potential to become a reliable alternative for quantitative studies of biomolecular interactions. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  13. Maternally derived trypsin may have multiple functions in the early development of turbot (Scopthalmus maximus).

    Science.gov (United States)

    Chi, Liang; Liu, Qinghua; Xu, Shihong; Xiao, Zhizhong; Ma, Daoyuan; Li, Jun

    2015-10-01

    Trypsin is an important serine protease that is considered to be involved in digestion of protein in teleost fish. Nevertheless, studies on trypsin/trypsinogen in fish embryos are very limited. In this study, the trypsinogen of turbot (Scophthalmus maximus) (tTG) was identified and the expression patterns and activity of trypsinogen/trypsin were investigated. The results showed that the tTG mRNA was evenly distributed in the oocytes and was also expressed along the yolk periphery in early embryos. At later embryo stages and 1 days after hatching (dph), the tTG mRNA concentrated at the alimentary tract and head. Quantitative expression analysis showed that the tTG transcripts decreased after fertilization until the gastrula stage, then increased with the embryo and larvae development. This result was also confirmed by the specific activity analysis of trypsin and in-situ-hybridization (ISH). All of the results indicated that tTG in early embryo stages was maternally derived and expressed by itself after gastrula stages. Additionally, location of tTG mRNA in embryos and larvae was investigated; we considered that trypsin may have multiple functions during the embryo development process. Based on our results regarding trypsinogen in embryos and early development, we concluded that the trypsin/trypsinogen in turbot embryos was inherited from a maternal source and we suggested that trypsin in early development has multiple functions in the process of development. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Purification and characterization of a trypsin inhibitor from the seeds of Artocarpus heterophyllus Lam.

    Science.gov (United States)

    Lyu, Junchen; Liu, Yuan; An, Tianchen; Liu, Yujun; Wang, Manchuriga; Song, Yanting; Zheng, Feifei; Wu, Dan; Zhang, Yingxia; Deng, Shiming

    2015-05-01

    A proteinaceous inhibitor against trypsin was isolated from the seeds of Artocarpus heterophyllus Lam. by successive ammonium sulfate precipitation, ion-exchange, and gel-filtration chromatography. The trypsin inhibitor, named as AHLTI (A. heterophyllus Lam. trypsin inhibitor), consisted of a single polypeptide chain with a molecular weight of 28.5 kDa, which was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel-filtration chromatography. The N-terminal sequence of AHLTI was DEPPSELDAS, which showed no similarity to other known trypsin inhibitor sequence. AHLTI completely inhibited bovine trypsin at a molar ratio of 1:2 (AHLTI:trypsin) analyzed by native polyacrylamide gel electrophoresis, inhibition activity assay, and gel-filtration chromatography. Moreover, kinetic enzymatic studies were carried out to understand the inhibition mechanism of AHLTI against trypsin. Results showed that AHLTI was a competitive inhibitor with an equilibrium dissociation constant (Ki) of 3.7 × 10(-8) M. However, AHLTI showed weak inhibitory activity toward chymotrypsin and elastase. AHLTI was stable over a broad range of pH 4-8 and temperature 20-80°C. The reduction agent, dithiothreitol, had no obvious effect on AHLTI. The trypsin inhibition assays of AHLTI toward digestive enzymes from insect pest guts in vitro demonstrated that AHLTI was effective against enzymes from Locusta migratoria manilensis (Meyen). These results suggested that AHLTI might be a novel trypsin inhibitor from A. heterophyllus Lam. belonging to Kunitz family, and play an important role in protecting from insect pest. © The Author 2015. Published by ABBS Editorial Office in association with Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

  15. Framework for interpretation of trypsin-antitrypsin imbalance and genetic heterogeneity in pancreatitis.

    Science.gov (United States)

    Lin, Kun; Gao, Feng; Chen, Qingquan; Liu, Qicai; Chen, Shu

    2015-01-01

    Early intracellular premature trypsinogen activation was interpreted as the key initiator of pancreatitis. When the balance in the homeostasis of trypsin and antitrypsin system is disequilibrated, elevated aggressive enzymes directly attack the pancreatic tissue, which leads to pancreatic destruction and inflammation. However, trypsin alone is not enough to cause complications in pancreatitis, which may play a crucial role in modulating signaling events in the initial phase of the disease. NFκB activation is the major inflammatory pathway involved in the occurrence and development of pancreatitis and it can be induced by intrapancreatic activation of trypsinogen. Synthesis of trypsinogen occurs in endoplasmic reticulum (ER), and ER stress is an important early acinar cell event. Components of ER stress response are known to be able to trigger cell death as well as NFκB signaling cascade. The strongest evidence supporting the trypsin-centered theory is that gene mutations, which lead to the generation of more trypsin, or reduce the activity of trypsin inhibitors or trypsin degradation, are associated with pancreatitis. Thus, trypsin-antitrypsin imbalance may be the first step leading to pancreatic autodigestion and inducing other pathways. Continued experimental studies are necessary to determine the specific relationships between trypsin-antitrypsin imbalance and genetic heterogeneity in pancreatitis. In this article, we review the latest advances that contributed to the understanding of the basic mechanisms behind the occurrence and development of pancreatitis with a focus on the interpretation of trypsin-antitrypsin imbalance and their relationships with other inflammation pathways. We additionally highlight genetic predispositions to pancreatitis and possible mechanisms associated with them.

  16. Trypsin-specific Inhibitors from the Macrolepiota procera, Armillaria mellea and Amanita phalloides wild mushrooms.

    Science.gov (United States)

    Lukanc, Tjaša; Brzin, Jože; Kos, Janko; Sabotič, Jerica

    2017-01-01

    Wild growing mushrooms are a rich source of novel proteins with unique features. We have isolated and characterized trypsin inhibitors from two edible mushrooms, the honey fungus (Armillaria mellea) and the parasol mushroom (Macrolepiota procera), and from the poisonous death cap (Amanita phalloides). The trypsin inhibitors isolated: armespin, macrospin and amphaspin, have similar molecular masses, acidic isoelectric points and are not N-glycosylated. They are very strong trypsin inhibitors and weak chymotrypsin inhibitors. They are resistant to exposure to high temperatures and withstand extreme pH values. These exceptional characteristics are advantageous for their potential use in biotechnology, agriculture and medicine.

  17. Effects of trypsin on cellular, chromosomal and DNA damage induced by X-rays

    Science.gov (United States)

    Sprunt, Elizabeth A.

    When cells are trypsinized before irradiation, potentiation of cell killing is seen; this is known as the 'trypsin effect'. The trypsin effect is re-examined here in the light of experiments in which enzymatic modifications of DNA in permeabilized cells has become a powerful experimental tool (Bryant et al, 1978, Ahnstrom and Bryant,1982; Natarajan et al, 1980; Bryant, 1984, 1985; Natarajan and Obe, 1984) and where in some cases it is suspected that trypsinization as part of the technique could significantly alter cell membrane permeability and chromatin structure (Obe et al, 1985; Obe and Winkel, 1985; Bryant and Christie, 1989). The trypsin effect was investigated at various cellular levels, assaying for cell survival (to verify the potentiation), anaphase chromosomal aberrations, DNA damage and repair and lastly using a nucleoid assay to investigate the effect of trypsin on DNA-nuclear matrix interactions. Each of these are considered in separate chapters as individual studies, then all compared in the final discussion. A small potentiation effect of X-ray damage on cell killing was seen when using Chinese Hamster Ovary (CHO) cells but no potentiating effect was found in the murine Ehrlich ascites tumour (EAT) cell line. Trypsinization was found to increase the number of X-ray induced chromosomal anaphase abnormalities in EAT cells. To investigate the possibility that the basis of the trypsin effect lies in its action at the DNA level, further experiments were performed to monitor DNA damage and repair using the DNA unwinding and neutral elution techniques. No difference was seen in the unwinding kinetics or in the DNA unwinding dose-effect curves for induction of DNA single strand breakage (ssb); when using neutral elution however. treatment of cells with trypsin or buffer alone increased the incidence of X-ray induced double strand breaks (dsb) at higher doses. Trypsinized EAT cells were found to repair ssb after 12 Gy less rapidly than those treated with

  18. Mating-increases trypsin in female Drosophila hemolymph.

    Science.gov (United States)

    Pilpel, Noam; Nezer, Ifat; Applebaum, Shalom W; Heifetz, Yael

    2008-03-01

    Male-derived accessory gland proteins (Acps) are transferred to the female reproductive tract during mating and affect female reproductive maturation and behavior. Some Acps subsequently enter the female hemolymph. We hypothesized that humoral proteases are the primary effectors of Acp bioactivity by processing (activating) and/or degrading them. To test this hypothesis we examined the fate of one Acp, Drosophila melanogaster Sex Peptide (Acp70A, DrmSP), which possesses several putative serine-protease cleavage sites, in hemolymph of unmated and mated females. In D. melanogaster, DrmSP induces post-mating non-receptivity and enhances oogenesis. To determine if serine proteases regulate the duration of DrmSP activity in mated females, we performed kinetic analysis of cleavage of a synthetic N-terminal truncated DrmSP(8-36) (T-SP) with hemolymph of unmated versus mated females. We found that T-SP is cleaved more rapidly and completely in mated female hemolymph. Using LC-MS/MS analyses, we identified its primary cleavage sites, indicating that trypsin was the major endopeptidase regulating T-SP in hemolymph. This was verified in vitro by utilizing specific chromogenic serine-protease substrates and inhibitors. We propose that post-mating cleavage of DrmSP in the female hemolymph regulates the duration of the rapidly induced post-mating responses in D. melanogaster and that this is a specific example of Acp bioactivity regulated by hemolymph serine proteases.

  19. Investigation of the interactions of lysozyme and trypsin with biphenol A using spectroscopic methods

    Science.gov (United States)

    Wang, Yan-Qing; Chen, Ting-Ting; Zhang, Hong-Mei

    2010-03-01

    The interaction between bisphenol A (BPA) and lysozyme (or trypsin) was investigated by UV-vis absorption, fluorescence, synchronous fluorescence, and three-dimensional fluorescence spectra techniques under physiological pH 7.40. BPA effectively quenched the intrinsic fluorescence of lysozyme and trypsin via static quenching. H-bonds and van der Waals interactions played a major role in stabilizing the BPA-proteinase complex. The distance r between donor and acceptor was obtained to be 1.65 and 2.26 nm for BPA-lysozyme and BPA-trypsin complexes, respectively. The effect of BPA on the conformation of lysozyme and trypsin was analyzed using synchronous fluorescence and three-dimensional fluorescence spectra.

  20. The use of poly(ethylene terephthalate)-poly(aniline) composite for trypsin immobilisation

    Energy Technology Data Exchange (ETDEWEB)

    Caramori, S.S. [Laboratorio de Quimica de Proteinas, Departamento de Bioquimica e Biologia Molecular, Instituto de Ciencias Biologicas, Universidade Federal de Goias, Cx. Postal 131, 74001-970 Goiania-GO (Brazil)], E-mail: samanthabio@hotmail.com; Fernandes, K.F. [Laboratorio de Quimica de Proteinas, Departamento de Bioquimica e Biologia Molecular, Instituto de Ciencias Biologicas, Universidade Federal de Goias, Cx. Postal 131, 74001-970 Goiania-GO (Brazil)], E-mail: katia@icb.ufg.br

    2008-08-01

    This paper presents trypsin immobilisation on strips of poly(ethylene terephthalate)-poly(aniline), activated with glutaraldehyde (PET-PANIG) composite. The photomicrography of the material showed changes corresponding to the chemical modifications produced in the steps of synthesis. The immobilisation process was very efficient under optimal conditions (18.6%). The immobilised and free enzyme presented the same pH and temperature optimum. PET-PANIG-trypsin was able to hydrolyse casein, albumin, gelatine, and skimmed milk. Km{sub app} value for PET-PANIG-trypsin was very close to Km of the free enzyme for casein. Immobilised trypsin showed higher stability than the free enzyme, with 100% activity after 14 days of storage at 4 deg. C and 100% operational stability after 4 cycles of use.

  1. Influence of Different Genotypes on Trypsin Inhibitor Levels and Activity in Soybeans

    Directory of Open Access Journals (Sweden)

    Viktor A. Nedovic

    2007-01-01

    Full Text Available This study describes the relationship between the two major trypsin inhibitors (TI in soybean, i.e., the Kunitz (KTI and Bowman-Birk (BBI trypsin inhibitors, as well as between them and the corresponding trypsin inhibitor activity (TIA. Twelve investigated soybean genotypes showed significant differences in TI levels and TIA. A very strong positive correlation was found between the levels of KTI and total BBI (r = 0.94, P < 0.05. No relationship was found between KTI, BBI or total TI and TIA. Based on this data, it appears that the levels of major TI in soybean are related. Understanding the relationship between trypsin inhibitors and their activities could be useful for further improvement of the health impacts of soy proteins.

  2. Bioconjugation of trypsin onto gold nanoparticles: Effect of surface chemistry on bioactivity

    Energy Technology Data Exchange (ETDEWEB)

    Hinterwirth, Helmut; Lindner, Wolfgang [Department of Analytical Chemistry, University of Vienna, Waehringerstrasse 38, 1090 Vienna (Austria); Laemmerhofer, Michael, E-mail: michael.laemmerhofer@uni-tuebingen.de [Department of Analytical Chemistry, University of Vienna, Waehringerstrasse 38, 1090 Vienna (Austria)

    2012-07-06

    Highlights: Black-Right-Pointing-Pointer Size and spacer affect bioactivity of nanoparticulate trypsin reactor. Black-Right-Pointing-Pointer Increase of GNP's size increases activity of bound trypsin. Black-Right-Pointing-Pointer Increase of spacer length increases amount and activity of immobilized enzyme by factor 6. Black-Right-Pointing-Pointer Decrease of digestion time up to less than 1 h when trypsin immobilized onto GNPs. Black-Right-Pointing-Pointer Reduced auto-digestion compared to trypsin in-solution. - Abstract: The systematic study of activity, long-time stability and auto-digestion of trypsin immobilized onto gold nanoparticles (GNPs) is described in this paper and compared to trypsin in-solution. Thereby, the influence of GNP's size and immobilization chemistry by various linkers differing in lipophilicity/hydrophilicity and spacer lengths was investigated with regard to the bioactivity of the conjugated enzyme. GNPs with different sizes were prepared by reduction and simultaneous stabilization with trisodium citrate and characterized by UV/vis spectra, dynamic light scattering (DLS), {zeta}-potential measurements and transmission electron microscopy (TEM). GNPs were derivatized by self-assembling of bifunctional thiol reagents on the nanoparticle (NP) surface via dative thiol-gold bond yielding a carboxylic acid functionalized surface. Trypsin was either attached directly via hydrophobic and ionic interactions onto the citrate stabilized GNPs or immobilized via EDC/NHS bioconjugation onto the carboxylic functionalized GNPs, respectively. The amount of bound trypsin was quantified by measuring the absorbance at 280 nm. The activity of bound enzyme and its Michaelis Menten kinetic parameter K{sub m} and v{sub max} were measured by the standard chromogenic substrate N{sub {alpha}}-Benzoyl-DL-arginine 4-nitroanilide hydrochloride (BApNA). Finally, digestion of a standard protein mixture with the trypsin-conjugated NPs followed by analysis with

  3. FRET-based modified graphene quantum dots for direct trypsin quantification in urine.

    Science.gov (United States)

    Poon, Chung-Yan; Li, Qinghua; Zhang, Jiali; Li, Zhongping; Dong, Chuan; Lee, Albert Wai-Ming; Chan, Wing-Hong; Li, Hung-Wing

    2016-04-21

    A versatile nanoprobe was developed for trypsin quantification with fluorescence resonance energy transfer (FRET). Here, fluorescence graphene quantum dot is utilized as a donor while a well-designed coumarin derivative, CMR2, as an acceptor. Moreover, bovine serum albumin (BSA), as a protein model, is not only served as a linker for the FRET pair, but also a fluorescence enhancer of the quantum dots and CMR2. In the presence of trypsin, the FRET system would be destroyed when the BSA is digested by trypsin. Thus, the emission peak of the donor is regenerated and the ratio of emission peak of donor/emission peak of acceptor increased. By the ratiometric measurement of these two emission peaks, trypsin content could be determined. The detection limit of trypsin was found to be 0.7 μg/mL, which is 0.008-fold of the average trypsin level in acute pancreatitis patient's urine suggesting a high potential for fast and low cost clinical screening. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Dissection of the binding of hydrogen peroxide to trypsin using spectroscopic methods and molecular modeling

    Science.gov (United States)

    Song, Wei; Yu, Zehua; Hu, Xinxin; Liu, Rutao

    2015-02-01

    Studies on the effects of environmental pollutants to protein in vitro has become a global attention. Hydrogen peroxide (H2O2) is used as an effective food preservative and bleacher in industrial production. The toxicity of H2O2 to trypsin was investigated by multiple spectroscopic techniques and the molecular docking method at the molecular level. The intrinsic fluorescence of trypsin was proved to be quenched in a static process based on the results of fluorescence lifetime experiment. Hydrogen bonds interaction and van der Waals forces were the main force to generate the trypsin-H2O2 complex on account of the negative ΔH0 and ΔS0. The binding of H2O2 changed the conformational structures and internal microenvironment of trypsin illustrated by UV-vis absorption, fluorescence, synchronous fluorescence, three-dimensional (3D) fluorescence and circular dichroism (CD) results. However, the binding site was far away from the active site of trypsin and the trypsin activity was only slightly affected by H2O2, which was further explained by molecular docking investigations.

  5. A trypsin inhibitor from Sapindus saponaria L. seeds: purification, characterization, and activity towards pest insect digestive enzyme.

    Science.gov (United States)

    Macedo, Maria Lígia R; Diz Filho, Eduardo B S; Freire, Mariadas Graças M; Oliva, Maria Luiza V; Sumikawa, Joana T; Toyama, Marcos H; Marangoni, Sérgio

    2011-01-01

    The present paper describes the purification, characterization and determination of the partial primary structure of the first trypsin inhibitor isolated from the family Sapindaceae. A highly stable, potent trypsin inhibitor (SSTI) was purified to homogeneity. SDS-PAGE analysis revealed that the protein consists of a two-polypeptide chain with molecular masses of approximately 15 and 3 kDa. The purified inhibitor inhibited bovine trypsin at a 1:1 M ratio. Kinetic analysis revealed that the protein is a competitive inhibitor with an equilibrium dissociation constant of 10⁻⁹ M for trypsin. The partial NH₂- terminal sequence of 36 amino acids in SSTI indicates homology with other members of the trypsin-inhibitor family from different sources. This inhibitor is highly stable in the presence of denaturing agents. SSTI showed significant inhibitory activity against trypsin-like proteases present in the larval midgut on Anagasta kuehniella, Corcyra cephalonica, Diatreae saccharalis and Anticarsia gemmatalis.

  6. Maize (Zea mays)-derived bovine trypsin: characterization of the first large-scale, commercial protein product from transgenic plants.

    Science.gov (United States)

    Woodard, Susan L; Mayor, Jocelyne M; Bailey, Michele R; Barker, Donna K; Love, Robert T; Lane, Jeffrey R; Delaney, Donna E; McComas-Wagner, Janet M; Mallubhotla, Hanuman D; Hood, Elizabeth E; Dangott, Lawrence J; Tichy, Shane E; Howard, John A

    2003-10-01

    Bovine trypsin (EC 3.4.21.4) is an enzyme that is widely used for commercial purposes to digest or process other proteins, including some therapeutic proteins. The biopharmaceutical industry is trying to eliminate animal-derived proteins from manufacturing processes due to the possible contamination of these products by human pathogens. Recombinant trypsin has been produced in a number of systems, including cell culture, bacteria and yeast. To date, these expression systems have not produced trypsin on a scale sufficient to fulfill the need of biopharmaceutical manufacturers where kilogram quantities are often required. The present paper describes commercial-level production of trypsin in transgenic maize (Zea mays) and its physical and functional characterization. This protease, the first enzyme to be produced on a large-scale using transgenic plant technology, is functionally equivalent to native bovine pancreatic trypsin. The availability of this reagent should allow for the replacement of animal-derived trypsin in the processing of pharmaceutical proteins.

  7. Recognizing Body Dysmorphic Disorder (Dysmorphophobia).

    Science.gov (United States)

    Varma, Anukriti; Rastogi, Rajesh

    2015-01-01

    Dysmorphophobia is a psychiatric condition which frequently presents in the clinics of dermatologists and plastic surgeons. This disorder (also called body dysmorphic disorder) is troublesome to the patient whilst being confusing for the doctor. This commonly undiagnosed condition can be detected by a few simple steps. Timely referral to a psychiatrist benefits most patients suffering from it. This article describes with a case vignette, how to recognize body dysmorphic disorder presenting in the dermatological or aesthetic surgery set up. Diagnostic criteria, eitiology, approach to patient, management strategy and when to refer are important learning points. The importance of recognizing this disorder timely and referring the patient to the psychiatrist for appropriate treatment is crucial. This article covers all aspects of body dysmorphic disorder relevant to dermatologists and plastic surgeons and hopes to be useful in a better understanding of this disorder.

  8. Recognizing body dysmorphic disorder (dysmorphophobia

    Directory of Open Access Journals (Sweden)

    Anukriti Varma

    2015-01-01

    Full Text Available Dysmorphophobia is a psychiatric condition which frequently presents in the clinics of dermatologists and plastic surgeons. This disorder (also called body dysmorphic disorder is troublesome to the patient whilst being confusing for the doctor. This commonly undiagnosed condition can be detected by a few simple steps. Timely referral to a psychiatrist benefits most patients suffering from it. This article describes with a case vignette, how to recognize body dysmorphic disorder presenting in the dermatological or aesthetic surgery set up. Diagnostic criteria, eitiology, approach to patient, management strategy and when to refer are important learning points. The importance of recognizing this disorder timely and referring the patient to the psychiatrist for appropriate treatment is crucial.This article covers all aspects of body dysmorphic disorder relevant to dermatologists and plastic surgeons and hopes to be useful in a better understanding of this disorder.

  9. Investigate the binding of catechins to trypsin using docking and molecular dynamics simulation.

    Science.gov (United States)

    Cui, Fengchao; Yang, Kecheng; Li, Yunqi

    2015-01-01

    To explore the inhibitory mechanism of catechins for digestive enzymes, we investigated the binding mode of catechins to a typical digestive enzyme-trypsin and analyzed the structure-activity relationship of catechins, using an integration of molecular docking, molecular dynamics simulation and binding free energy calculation. We found that catechins with different structures bound to a conservative pocket S1 of trypsin, which is comprised of residues 189-195, 214-220 and 225-228. In the trypsin-catechin complexes, Asp189 by forming strong hydrogen bonding, and Gln192, Trp215 and Gly216 through hydrophobic interactions, all significantly contribute to the binding of catechins. The number and the position of hydroxyl and aromatic groups, the structure of stereoisomers, and the orientation of catechins in the binding pocket S1 of trypsin all affect the binding affinity. The binding affinity is in the order of Epigallocatechin gallate (EGCG) > Epicatechin gallate (ECG) > Epicatechin (EC) > Epigallocatechin (EGC), and 2R-3R EGCG shows the strongest binding affinity out of other stereoisomers. Meanwhile, the synergic conformational changes of residues and catechins were also analyzed. These findings will be helpful in understanding the knowledge of interactions between catechins and trypsin and referable for the design of novel polyphenol based functional food and nutriceutical formulas.

  10. Buckwheat trypsin inhibitor with helical hairpin structure belongs to a new family of plant defence peptides.

    Science.gov (United States)

    Oparin, Peter B; Mineev, Konstantin S; Dunaevsky, Yakov E; Arseniev, Alexander S; Belozersky, Mikhail A; Grishin, Eugene V; Egorov, Tsezi A; Vassilevski, Alexander A

    2012-08-15

    A new peptide trypsin inhibitor named BWI-2c was obtained from buckwheat (Fagopyrum esculentum) seeds by sequential affinity, ion exchange and reversed-phase chromatography. The peptide was sequenced and found to contain 41 amino acid residues, with four cysteine residues involved in two intramolecular disulfide bonds. Recombinant BWI-2c identical to the natural peptide was produced in Escherichia coli in a form of a cleavable fusion with thioredoxin. The 3D (three-dimensional) structure of the peptide in solution was determined by NMR spectroscopy, revealing two antiparallel α-helices stapled by disulfide bonds. Together with VhTI, a trypsin inhibitor from veronica (Veronica hederifolia), BWI-2c represents a new family of protease inhibitors with an unusual α-helical hairpin fold. The linker sequence between the helices represents the so-called trypsin inhibitory loop responsible for direct binding to the active site of the enzyme that cleaves BWI-2c at the functionally important residue Arg(19). The inhibition constant was determined for BWI-2c against trypsin (1.7×10(-1)0 M), and the peptide was tested on other enzymes, including those from various insect digestive systems, revealing high selectivity to trypsin-like proteases. Structural similarity shared by BWI-2c, VhTI and several other plant defence peptides leads to the acknowledgement of a new widespread family of plant peptides termed α-hairpinins.

  11. Crystallization, data collection and processing of the chymotrypsin–BTCI–trypsin ternary complex

    Energy Technology Data Exchange (ETDEWEB)

    Esteves, Gisele Ferreira; Teles, Rozeni Chagas Lima; Cavalcante, Nayara Silva; Neves, David; Ventura, Manuel Mateus [Laboratório de Biofísica, Instituto de Ciências Biológicas, Universidade de Brasília, 70910-900 Brasília-DF (Brazil); Barbosa, João Alexandre Ribeiro Gonçalves, E-mail: joao@lnls.br [Center for Structural Molecular Biology (CeBiME), Brazilian Synchrotron Light Laboratory (LNLS), CP 6192, 13083-970 Campinas-SP (Brazil); Freitas, Sonia Maria de, E-mail: joao@lnls.br [Laboratório de Biofísica, Instituto de Ciências Biológicas, Universidade de Brasília, 70910-900 Brasília-DF (Brazil)

    2007-12-01

    A ternary complex of the proteinase inhibitor (BTCI) with trypsin and chymotrypsin was crystallized and its crystal structure was solved by molecular replacement. A ternary complex of the black-eyed pea trypsin and chymotrypsin inhibitor (BTCI) with trypsin and chymotrypsin was crystallized by the sitting-drop vapour-diffusion method with 0.1 M HEPES pH 7.5, 10%(w/v) polyethylene glycol 6000 and 5%(v/v) 2-methyl-2,4-pentanediol as precipitant. BTCI is a small protein with 83 amino-acid residues isolated from Vigna unguiculata seeds and is able to inhibit trypsin and chymotrypsin simultaneously by forming a stable ternary complex. X-ray data were collected from a single crystal of the trypsin–BTCI–chymotrypsin ternary complex to 2.7 Å resolution under cryogenic conditions. The structure of the ternary complex was solved by molecular replacement using the crystal structures of the BTCI–trypsin binary complex (PDB code) and chymotrypsin (PDB code) as search models.

  12. TRYPSIN-INDUCED HEMAGGLUTINATION ASSAY FOR THE DETECTION OF INFECTIOUS BRONCHITIS VIRUS

    Directory of Open Access Journals (Sweden)

    M. S. Mahmood, M. Siddique, I. Hussain and A. Khan1

    2004-04-01

    Full Text Available A trypsin-induced hemagglutination (THA assay was standardized to detect infectious bronchitis virus (IBV in allantoic fluid (AF of embryonated eggs. The test was used in 20 samples, each collected from 5 different layer farms suspected for IBV. Allantoic fluid from inoculated embryos was harvested and treated with reagent grade trypsin at the percentages of 0.25, 0.50, 1.0 and 2.0 for 30 minutes to 3 hours at pH 7.2. The IBV in trypsinized AF was identified by clear and consistent agglutination of chicken red blood cells within 5 minutes of incubation at 37oC. The results indicated that AF treated with equal volume of 1.0% reagent grade trypsin elicited the hemagglutinating (HA activity in 3.0 hours whereas 2.0% reagent grade trypsin elicited the HA activity only after 30 minutes incubation at 37oC. Sensitivity of THA was 92% as compared with 76% for agar gel precipitation test. Gross pathological lesions (curling and dwarfing in chick embryo, intracereberal inoculation of un-weaned mice and pathogenicity test in one-day-old broiler chicks showed 79, 84 and 77% sensitivity, respectively.

  13. Trypsin inhibitor from tamarindus indica L. seeds reduces weight gain and food consumption and increases plasmatic cholecystokinin levels

    Directory of Open Access Journals (Sweden)

    Joycellane Alline do Nascimento Campos Ribeiro

    2015-02-01

    Full Text Available OBJECTIVES: Seeds are excellent sources of proteinase inhibitors, some of which may have satietogenic and slimming actions. We evaluated the effect of a trypsin inhibitor from Tamarindus indica L. seeds on weight gain, food consumption and cholecystokinin levels in Wistar rats. METHODS: A trypsin inhibitor from Tamarindus was isolated using ammonium sulfate (30-60% following precipitation with acetone and was further isolated with Trypsin-Sepharose affinity chromatography. Analyses were conducted to assess the in vivo digestibility, food intake, body weight evolution and cholecystokinin levels in Wistar rats. Histological analyses of organs and biochemical analyses of sera were performed. RESULTS: The trypsin inhibitor from Tamarindus reduced food consumption, thereby reducing weight gain. The in vivo true digestibility was not significantly different between the control and Tamarindus trypsin inhibitor-treated groups. The trypsin inhibitor from Tamarindus did not cause alterations in biochemical parameters or liver, stomach, intestine or pancreas histology. Rats treated with the trypsin inhibitor showed significantly elevated cholecystokinin levels compared with animals receiving casein or water. CONCLUSION: The results indicate that the isolated trypsin inhibitor from Tamarindus reduces weight gain by reducing food consumption, an effect that may be mediated by increased cholecystokinin. Thus, the potential use of this trypsin inhibitor in obesity prevention and/or treatment should be evaluated.

  14. Molecular mechanism of interaction between norfloxacin and trypsin studied by molecular spectroscopy and modeling

    Science.gov (United States)

    Lu, Yan; Wang, Gongke; Lu, Xiumin; Lv, Juan; Xu, Meihua; Zhang, Weiwei

    2010-01-01

    The molecular mechanism of the binding of norfloxacin (NRF) to trypsin was investigated by fluorescence, synchronous fluorescence and UV-vis absorbance spectroscopy and molecular modeling at physiological conditions. The quenching mechanism and the binding mode were investigated in terms of the association constants and basic thermodynamic parameters. The results of spectroscopic measurements suggested that NRF have a strong ability to quench the intrinsic fluorescence of trypsin through static quenching procedure. Moreover, fluorescence experiments were also performed at different values of pH to elucidate the effect of pH on the binding. The NRF-trypsin complex was stabilized by hydrophobic forces and hydrogen bonding, via tryptophan residue as indicated from the thermodynamic parameters, which was consistent with the results of molecular docking and accessible surface area calculations.

  15. The effects of nitrogen on protein, oil and trypsin inhibitor content of soybean

    Directory of Open Access Journals (Sweden)

    Perić Vesna

    2009-01-01

    Full Text Available Nitrogen fertilization have influence on protein, oil and trypsin inhibitor content of different soybean genotypes. Seed protein content was increased over control by 60 kg ha-1 nitrogen while trypsin inhibitor was reduced by all treatmens (30, 60,90 N kg ha-1 as compared to controls. Significant genetic variation in TI was found both within the genotype class with the Kunitz inhibitor present as well as within the class lacking this inhibitor. Genotypes containing the Kunitz trypsin inhibitor protein (KTI exhibit a higher TI than genotypes lacking this protein, however, in both groups of genotypes TI was similary affected by nitrogen application. Oil content was reduced following nitrogen fertilisation.

  16. [Recognizing signs of sexual abuse

    NARCIS (Netherlands)

    Teunissen, T.A.; Wong, S.H.; Lagro-Janssen, A.L.

    2016-01-01

    Sexual abuse is very common. In the Netherlands 42% of women and 13% of men aged over 25 years have experienced unacceptable sexual behaviour. Most victims do not seek professional help nor do they report the abuse to the police, and most of the victims who do seek medical help do not mention the

  17. Trypsin Reduces Pancreatic Ductal Bicarbonate Secretion by Inhibiting CFTR Cl- channel and Luminal Anion Exchangers

    Science.gov (United States)

    Pallagi, Petra; Venglovecz, Viktória; Rakonczay, Zoltán; Borka, Katalin; Korompay, Anna; Ózsvári, Béla; Judák, Linda; Sahin-Tóth, Miklós; Geisz, Andrea; Schnúr, Andrea; Maléth, József; Takács, Tamás; Gray, Mike A.; Argent, Barry E.; Mayerle, Julia; Lerch, Markus M.; Wittmann, Tibor; Hegyi, Péter

    2012-01-01

    Background & Aims The effects of trypsin on pancreatic ductal epithelial cells (PDEC) vary among species and depend on localization of proteinase-activated receptor-2 (PAR-2). Bicarbonate secretion is similar in human and guinea pig PDEC; we compared its localization in these cell types and isolated guinea pig ducts to study the effects of trypsin and a PAR-2 agonist on this process. Methods PAR-2 localization was analyzed by immunohistochemistry in guinea pig and human pancreatic tissue samples (from 15 patients with chronic pancreatitis and 15 without pancreatic disease). Functions of guinea pig PDEC were studied by microperfusion of isolated ducts, measurements of intracellular pH (pHi) and Ca2+ concentration [Ca2+]i, and patch clamp analysis. The effect of pH on trypsinogen autoactivation was assessed using recombinant human cationic trypsinogen. Results PAR-2 localized to the apical membrane of human and guinea pig PDEC. Trypsin increased [Ca2+]i and pHi, and inhibited secretion of bicarbonate by the luminal anion exchanger and the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel. Autoactivation of human cationic trypsinogen accelerated when the pH was reduced from 8.5 to 6.0. PAR-2 expression was strongly down-regulated, at transcriptional and protein levels, in the ducts of patients with chronic pancreatitis, consistent with increased activity of intraductal trypsin. Importantly, in PAR-2 knockout mice, the effects of trypsin were PAR-2 dependent. Conclusions Trypsin reduces pancreatic ductal bicarbonate secretion via PAR-2–dependent inhibition of the apical anion exchanger and the CFTR Cl- channel. This could contribute to the development of chronic pancreatitis, decreasing luminal pH and promoting premature activation of trypsinogen in the pancreatic ducts. PMID:21893120

  18. The adsorption of trypsin on hydrophilic and hydrophobic surfaces. In situ structural characterization of the enzyme in the adsorbed state

    NARCIS (Netherlands)

    Koutsopoulos, S.; Patzsch, K.; Bosker, W.T.E.; Norde, W.

    2007-01-01

    The adsorption of trypsin onto polystyrene and silica surfaces was investigated by reflectometry, spectroscopic methods, and atomic force microscopy (AFM). The affinity of trypsin for the hydrophobic polystyrene surface was higher than that for the hydrophilic silica surface, but steady-state

  19. Molecular characterization of genes encoding trypsin-like enzymes from Aedes aegypti larvae and identification of digestive enzymes.

    Science.gov (United States)

    Soares, Tatiane S; Watanabe, Renata M O; Lemos, Francisco J A; Tanaka, Aparecida S

    2011-12-10

    Trypsin-like enzymes play an important role in the Aedes aegypti digestive process. The trypsin-like enzymes present in adults were characterized previously, but little is known about trypsins in larvae. In the present work, we identified one of the trypsin enzymes from Ae. aegypti larval midgut using a library of trypsin gene fragments, which was the sequence known as AAEL005607 from the Ae. aegypti genome. Quantitative PCR analysis showed that AAEL005607 was transcribed in all larval instars, but it was not present in adult midgut. In order to confirm transcription data, the trypsin-like enzymes from 4th instar larvae of Ae. aegypti midgut were purified and sequenced. Purified trypsin showed identity with the amino-terminal sequence of AAEL005607, AAEL005609 and AAEL005614. These three trypsins have high amino acids identity, and could all be used as a template for the design of inhibitors. In conclusion, for the first time, digestive enzymes of 4th larval instar of Ae. aegypti were purified and characterized. The knowledge of digestive enzymes present in Ae. aegypti larvae may be helpful in the development of a larvicide. Copyright © 2011 Elsevier B.V. All rights reserved.

  20. Flow Cytometry Method Analysis of Apoptosis: No Significant Difference Between EDTA and EDTA-free Trypsin Treatment Procedure.

    Science.gov (United States)

    Xu, Xiao-yan; Nie, Xiao-cui; Ma, Hai-ying; Song, Guo-qing; Zhang, Xiao-tong; Jin, Yu-nan; Yu, Yan-qiu

    2015-04-01

    Flow cytometry method (FCM) is a generally accepted tool to analyze apoptosis. Although apoptosis assay kit was applied by many companies, the manufacturers were not consistent with whether using Trypsin with EDTA to collect the adherent cells. In another words, the influence of EDTA on apoptotic ratio is not clear. In this work, we compared the proportion of apoptotic cells with EDTA or EDTA-free Trypsin treatment by FCM. We concluded that Trypsin with or without EDTA has little influence on the proportion of apoptotic cells. In addition, we found that the ratio of necrosis and apoptosis was different in cells collected by scraping. WAVE2 protein was analyzed as a typical example for movement related protein. WAVE2 expression is elevated in the EDTA Trypsin treated group, compared with EDTA-free Trypsin treatment and scrapping group. © The Author(s) 2014.

  1. Properties of purified gut trypsin from Helicoverpa zea, adapted to to proteinase inhibitors.

    NARCIS (Netherlands)

    Volpicella, M.; Ceci, L.R.; America, T.; Gallarani, R.; Bode, W.; Jongsma, M.A.; Beekwilder, J.

    2003-01-01

    Pest insects such as Helicoverpa spp. frequently feed on plants expressing protease inhibitors. Apparently, their digestive system can adapt to the presence of protease inhibitors. To study this, a trypsin enzyme was purified from the gut of insects that were raised on an inhibitor-containing diet.

  2. The quaternary structure of chicken acetylcholinesterase and butyrylcholinesterase; effect of collagenase and trypsin.

    Science.gov (United States)

    Allemand, P; Bon, S; Massoulié, J; Vigny, M

    1981-03-01

    Acetylcholinesterase (EC 3.1.1.7.; AChE) and butyrylcholinesterase (EC 3.1.1.8.; BuChE) from chicken muscle exist as sets of structurally homologous forms with very similar properties. The collagenase sensitivity and aggregation properties of the 'heavy' forms of both enzymes indicate that they possess a collagen-like tail, and their stepwise dissociation by trypsin confirms that they correspond to triple (A12) and double (A8) collagen-tailed tetramers. In addition to this dissociating effect, trypsin digests an important fraction of the catalytic units of AChE, in a progressive manner, removing as much as 30% of the enzyme's mass, without inactivation of the tetramers and of the tailed molecules. The trypsin-modified AChE forms closely resemble the corresponding mammalian AChE forms in their hydrodynamic properties. It is not known whether the trypsin-digestible peptides, which do not appear to be involved in the ionic or hydrophobic interactions of the enzymes, are a fragment of the catalytic subunit or whether they constitute distinct polypeptides.

  3. Proteolytic Activation of the Porcine Epidemic Diarrhea Coronavirus Spike Fusion Protein by Trypsin in Cell Culture.

    NARCIS (Netherlands)

    Wicht, Oliver|info:eu-repo/dai/nl/32291177X; Li, Wentao|info:eu-repo/dai/nl/411296272; Willems, Lione; Meuleman, Tom J; Wubbolts, Richard W|info:eu-repo/dai/nl/181688255; van Kuppeveld, Frank J M|info:eu-repo/dai/nl/156614723; Rottier, Peter J M|info:eu-repo/dai/nl/068451954; Bosch, Berend Jan|info:eu-repo/dai/nl/273306049

    2014-01-01

    Isolation of porcine epidemic diarrhea coronavirus (PEDV) from clinical material in cell culture requires supplementation of trypsin. This may relate to the confinement of PEDV natural infection to the protease-rich small intestine of pigs. Our study focused on the role of protease activity on

  4. Inactivation of soybean trypsin inhibitors and lipoxygenases by high-pressure processing

    NARCIS (Netherlands)

    Ven, van der C.; Matser, A.M.; Berg, van den R.W.

    2005-01-01

    Trypsin inhibitors (TIA), one of the antinutritional factors of soy milk, are usually inactivated by heat treatment. In the current study, high-pressure processing (HPP) was evaluated as an alternative for the inactivation of TIA in soy milk. Moreover, the effect of HPP on lipoxygenase (LOX) in

  5. Trypsin-mediated enzymatic degradation of type II collagen in the human vitreous

    NARCIS (Netherlands)

    van Deemter, Marielle; Kuijer, Roel; Pas, Hendri Harm; van der Worp, Roelofje Jacoba; Hooymans, Johanna Martina Maria; Los, Leonoor Inge

    2013-01-01

    Purpose: Aging of the vitreous body can result in sight-threatening pathology. One aspect of vitreous aging is liquefaction, which results from the vanishing of collagen fibrils. We investigated the possibility that trypsins are involved in vitreous type II collagen degradation. Methods:

  6. Trypsin inhibitors from Capsicum baccatum var. pendulum leaves involved in Pepper yellow mosaic virus resistance.

    Science.gov (United States)

    Moulin, M M; Rodrigues, R; Ribeiro, S F F; Gonçalves, L S A; Bento, C S; Sudré, C P; Vasconcelos, I M; Gomes, V M

    2014-11-07

    Several plant organs contain proteinase inhibitors, which are produced during normal plant development or are induced upon pathogen attack to suppress the enzymatic activity of phytopathogenic microorganisms. In this study, we examined the presence of proteinase inhibitors, specifically trypsin inhibitors, in the leaf extract of Capsicum baccatum var. pendulum inoculated with PepYMV (Pepper yellow mosaic virus). Leaf extract from plants with the accession number UENF 1624, which is resistant to PepYMV, was collected at 7 different times (0, 24, 48, 72, 96, 120, and 144 h). Seedlings inoculated with PepYMV and control seedlings were grown in a growth chamber. Protein extract from leaf samples was partially purified by reversed-phase chromatography using a C2/C18 column. Residual trypsin activity was assayed to detect inhibitors followed by Tricine-SDS-PAGE analysis to determine the N-terminal peptide sequence. Based on trypsin inhibitor assays, trypsin inhibitors are likely constitutively synthesized in C. baccatum var. pendulum leaf tissue. These inhibitors are likely a defense mechanism for the C. baccatum var. pendulum- PepYMV pathosystem.

  7. Purification and characterization of a trypsin inhibitor from seeds of Murraya koenigii.

    Science.gov (United States)

    Shee, Chandan; Sharma, Ashwani K

    2007-02-01

    A protein with trypsin inhibitory activity was purified to homogeneity from the seeds of Murraya koenigii (curry leaf tree) by ion exchange chromatography and gel filtration chromatography on HPLC. The molecular mass of the protein was determined to be 27 kDa by SDS-PAGE analysis under reducing conditions. The solubility studies at different pH conditions showed that it is completely soluble at and above pH 7.5 and slowly precipitates below this pH at a protein concentration of 1 mg/ml. The purified protein inhibited bovine pancreatic trypsin completely in a molar ratio of 1:1.1. Maximum inhibition was observed at pH 8.0. Kinetic studies showed that Murraya koenigii trypsin inhibitor is a competitive inhibitor with an equilibrium dissociation constant of 7 x 10(-9) M. The N-terminal sequence of the first 15 amino acids showed no similarity with any of the known trypsin inhibitors, however, a short sequence search showed significant homology to a Kunitz-type chymotrypsin inhibitor from Erythrina variegata.

  8. Effects of tannic acid on trypsin and leucine aminopeptidase activities in gypsy moth larval midgut

    Directory of Open Access Journals (Sweden)

    Mrdaković Marija

    2013-01-01

    Full Text Available The effects of allelochemical stress on genetic variations in the specific activities of gypsy moth digestive enzymes (trypsin and leucine aminopeptidase and relative midgut mass (indirect measure of food consumption, as well as variability in their plasticity, were investigated in fifth instar gypsy moths originating from two populations with different trophic adaptations (oak and locust-tree forests. Thirty-two full-sib families from the Quercus population and twenty-six full-sib families from the Robinia population were reared on an artificial diet with or without supplementation with tannic acid. Between population differences were observed as higher average specific activity of trypsin and relative midgut mass in larvae from the Robinia population. Significant broad-sense heritabilities were observed for the specific activity of trypsin in the control state, and for specific activity of leucine aminopeptidase in a stressful environment. Significantly lower heritability for relative midgut mass was recorded in larvae from the Robinia population reared under stressful conditions. Significant variability of trypsin plasticity in larvae from both populations and significant variability of leucine aminopeptidase plasticity in larvae from the Robinia population point to the potential for the evolution of enzyme adaptive plastic responses to the presence of stressor. Non-significant across-environment genetic correlations do not represent a constraint for the evolution of enzyme plasticity. [Projekat Ministarstva nauke Republike Srbije, br. 173027

  9. Trypsin-like proteins of the fungi as possible markers of pathogenicity.

    Science.gov (United States)

    Dubovenko, Aleksej G; Dunaevsky, Yakov E; Belozersky, Mikhail A; Oppert, Brenda; Lord, Jeffrey C; Elpidina, Elena N

    2010-01-01

    Sequences of peptidases with conserved motifs around the active site residues that are characteristic of trypsins (similar to trypsin peptidases, STP) were obtained from publicly-available fungal genomes and related databases. Among the 75 fungal genomes, 29 species of parasitic Ascomycota contained genes encoding STP and their homologs. Searches of non-redundant protein sequences, patented protein sequences, and expressed sequence tags resulted in another 18 STP sequences in 10 fungal species from Ascomycota, Basidiomycota, and Zygomycota. A comparison of fungi species containing STP sequences revealed that almost all are pathogens of plants, animals or fungi. A comparison of the primary structure of homologous proteins, including the residues responsible for substrate binding and specificity of the enzyme, revealed three groups of homologous sequences, all presumably from S1 family: trypsin-like peptidases, chymotrypsin-like peptidases and serine peptidases with unknown substrate specificity. Homologs that are presumably functionally inactive were predicted in all groups. The results in general support the hypothesis that the expression of trypsin-like peptidases in fungi represents a marker of fungal phytopathogenicity. A phylogenetic tree was constructed using peptidase and homolog amino acid sequences, demonstrating that all have noticeable differences and almost immediately deviate from the common root. Therefore, we conclude that the changes that occurred in STP of pathogenic fungi in the course of evolution represent specific adaptations to proteins of their respective hosts, and mutations in peptidase genes are important components of life-style changes and taxonomic divergence.

  10. Ecological costs and benefits correlated with trypsin protease inhibitor production in Nicotiana attenuata

    NARCIS (Netherlands)

    Glawe, G.A.; Zavala, J.A.; Kessler, A.; Van Dam, N.M.; Baldwin, I.T.

    2003-01-01

    Genotypes of the wild tobacco Nicotiana attenuata from different geographic regions in North America vary considerably in the level of constitutive and inducible trypsin protease inhibitors (TrypPIs), a potent direct defense, as well as in the production of herbivore-induced volatiles that function

  11. A randomized controlled trial of trypsin to treat brown recluse spider bites in Guinea pigs.

    Science.gov (United States)

    Cabaniss, Wyman W; Bush, Sean; O'Rourke, Dorcas P; Fletcher, Paul F; Brewer, Kori L; Lertpiriyapong, Kvin; Punja, Mohan; Miller, Susan N; Meggs, William J

    2014-09-01

    Brown recluse spider bites result in necrotic skin lesions for which there is no known antidote. Since venom toxins are proteins, a proteolytic enzyme like trypsin might be effective in reducing toxicity. The aim of this study was to conduct a randomized controlled trial of trypsin to treat brown recluse spider bites in guinea pigs. Subjects were 18 female guinea pigs. Anesthesia for injections was inhaled isoflurane. Analgesia was 0.05 mg/kg of buprenorphine twice a day as needed. Intervention was intradermal injection of 30 μg of brown recluse venom (Spider Pharm, Yarnell, AZ). Immediately after envenomation, subjects were randomized to two groups of nine: trypsin 10 μg in 1 mL normal saline and 1 mL of normal saline. The primary outcome was lesion area over a 10-day time period. Statistical analysis was performed with repeated measures ANOVA. Mean lesion area was smaller but not statistically different in the placebo group. Maximum lesion size occurred at day 4 in both groups, when lesion area was 76.1 ± 108.2 mm(2) in the placebo group and 149.7 ± 127.3 mm(2) in the treatment group. P value was 0.15 for placebo vs. treatment. This study did not establish a role for trypsin as a treatment for brown recluse spider bites in a guinea pig model.

  12. A NEW METHOD TO DETECT ACROSOME-REACTED SPERMATOZOA USING BIOTINYLATED SOYBEAN TRYPSIN-INHIBITOR

    NARCIS (Netherlands)

    ARTS, EGJM; KUIKEN, J; JAGER, S

    1994-01-01

    Objective: To develop a method to detect acrosome-reacted spermatozoa on human zonae pellucidae using only commercially available reagents and without need for sperm fixation. Design: Sperm head labeling with biotinylated soybean trypsin inhibitor (SBTI-biotin) was compared with results of a known

  13. Biochemical Characterization of An Arginine-Specific Alkaline Trypsin from Bacillus licheniformis

    Directory of Open Access Journals (Sweden)

    Jin-Song Gong

    2015-12-01

    Full Text Available In the present study, we isolated a trypsin-producing strain DMN6 from the leather waste and identified it as Bacillus licheniformis through a two-step screening strategy. The trypsin activity was increased up to 140 from 20 U/mL through culture optimization. The enzyme was purified to electrophoretic homogeneity with a molecular mass of 44 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the specific activity of purified enzyme is 350 U/mg with Nα-Benzoyl-l-arginine ethylester as the substrate. The optimum temperature and pH for the trypsin are 65 °C and pH 9.0, respectively. Also, the enzyme can be significantly activated by Ba2+. This enzyme is relatively stable in alkaline environment and displays excellent activity at low temperatures. It could retain over 95% of enzyme activity after 180 min of incubation at 45 °C. The distinguished activity under low temperature and prominent stability enhance its catalytic potential. In the current work, the open reading frame was obtained with a length of 1371 nucleotides that encoded a protein of 456 amino acids. These data would warrant the B. licheniformis trypsin as a promising candidate for catalytic application in collagen preparation and leather bating through further protein engineering.

  14. Interactive computer surface graphics approach to study of the active site of bovine trypsin

    Science.gov (United States)

    Feldmann, Richard J.; Bing, David H.; Furie, Barbara C.; Furie, Bruce

    1978-01-01

    A descriptive medium for the presentation of protein structure has been developed and used to evaluate the structure of the active site of bovine trypsin (EC 3.4.21.4). This technique, involving advanced computer graphics technology, permits the facile display of a representation of the molecular surface of proteins of known structure and employs color to code the structural or chemical features of this surface. Benzamidine derivatives were inserted into the benzamidine-binding site of trypsin and the binary inhibitor-trypsin complex was evaluated by using the computer-generated structure. On the basis of qualitative assessments of the contribution of electrostatic and hydrophobic forces to the binding energy associated with complex formation, we made predictions concerning the effects of interaction of benzamidine substituents and amino acid side chains upon the binding energy associated with inhibitor-protein binding. The computer display of the molecular surfaces of the binary complex of substituted benzamidines and trypsin permitted unique insight into the identity and chemical properties of the atoms that participate at the interface of the molecular surfaces of the inhibitor and the protein. The computer-generated molecular surface display can potentially be combined with quantitative definition of the physical forces involved in the interaction of molecular surfaces. This technology should facilitate the study of the structure-activity relationship of substrates, inhibitors, and drugs that bind to proteins of known three-dimensional structure. Images PMID:281690

  15. Toxic effects of Ricinus communis non-protein trypsin inhibitor on ...

    African Journals Online (AJOL)

    In the study reported herein, we aimed to isolate a trypsin inhibitor from Ricinus communis leaves through chromatographic and spectrometric techniques and evaluate its toxic effects on the development of Spodoptera frugiperda larvae. Plant extracts were submitted to fractionation in adsorption column. The fraction 10 ...

  16. Recombinant Buckwheat Trypsin Inhibitor Induces Mitophagy by Directly Targeting Mitochondria and Causes Mitochondrial Dysfunction in Hep G2 Cells.

    Science.gov (United States)

    Wang, Zhuanhua; Li, Shanshan; Ren, Rong; Li, Jiao; Cui, Xiaodong

    2015-09-09

    Mitochondria are essential targets for cancer chemotherapy and other disease treatments. Recombinant buckwheat trypsin inhibitor (rBTI), a member of the potato type I proteinase inhibitor family, was derived from tartary buckwheat extracts. Our results showed that rBTI directly targeted mitochondria and induced mitochondrial fragmentation and mitophagy. This occurs through enhanced depolarization of the mitochondrial membrane potential, increasing reactive oxygen species (ROS) generation associated with the rise of the superoxide dismutase and catalase activity and glutathione peroxidase (GSH) content, and changes in the GSH/oxidized glutathione ratio. Mild and transient ROS induced by rBTI were shown to be important signaling molecules required to induce Hep G2 mitophagy to remove dysfunctional mitochondria. Furthermore, rBTI could directly induce mitochondrial fragmentation. It was also noted that rBTI highly increased colocalization of mitochondria in treated cells compared to nontreated cells. Tom 20, a subunit of the translocase of the mitochondrial outer membrane complex responsible for recognizing mitochondrial presequences, may be the direct target of rBTI.

  17. Trypsin from the digestive system of carp Cirrhinus mrigala: purification, characterization and its potential application.

    Science.gov (United States)

    Khangembam, Bronson Kumar; Chakrabarti, Rina

    2015-05-15

    Trypsin was purified 35.64-fold with 4.97% recovery from the viscera of carp Cirrhinus mrigala (mrigal) by ammonium sulfate precipitation, ion exchange and affinity chromatography. The purified enzyme was active at a wide range of pH (7.0-9.2) and temperature (10-50°C). The purified enzyme exhibited high thermal stability up to 50°C for 1h. The enzyme activity was stabilized by Ca(+2) (2mM) up to 7h at 40°C. The Km and kcat values of purified enzyme were 0.0672 mM and 92.09/s/mM, respectively. Soybean trypsin inhibitor and phenylmethylsulphonylflouride completely inhibited the enzyme activity. The specific inhibitor of trypsin, N-α-p-tosyl-L-lysine chloromethyl ketone inhibited 99.67% activity. Na(+), K(+) and Li(+) inhibited 20.99 ± 5.25%, 16.53 ± 4.80% and 18.99 ± 1.42% of enzyme activity, respectively. Divalent ions Mg(+2), Zn(+2), Co(+2), Hg(+2) and Cd(+2) inhibited 21.61 ± 2.22%, 31.62 ± 1.78%, 31.62 ± 1.96%, 85.68 ± 1.51% and 47.95 ± 2.13% enzyme activity, respectively. SDS-PAGE showed that the molecular mass of purified enzyme was 21.7 kDa. MALDI-TOF study showed a peptide sequence of AFCGGSLVNENKMHSAGHCYKSRIQV at the N-Terminal. This sequence recorded 76-84% identity with trypsin from Thunnus thynnus and other fish species. This confirmed that the purified protein was trypsin. The purified enzyme has potential applications in detergent and food industry because of its thermal stability and alkaline nature. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Recognizing characters of ancient manuscripts

    Science.gov (United States)

    Diem, Markus; Sablatnig, Robert

    2010-02-01

    Considering printed Latin text, the main issues of Optical Character Recognition (OCR) systems are solved. However, for degraded handwritten document images, basic preprocessing steps such as binarization, gain poor results with state-of-the-art methods. In this paper ancient Slavonic manuscripts from the 11th century are investigated. In order to minimize the consequences of false character segmentation, a binarization-free approach based on local descriptors is proposed. Additionally local information allows the recognition of partially visible or washed out characters. The proposed algorithm consists of two steps: character classification and character localization. Initially Scale Invariant Feature Transform (SIFT) features are extracted which are subsequently classified using Support Vector Machines (SVM). Afterwards, the interest points are clustered according to their spatial information. Thereby, characters are localized and finally recognized based on a weighted voting scheme of pre-classified local descriptors. Preliminary results show that the proposed system can handle highly degraded manuscript images with background clutter (e.g. stains, tears) and faded out characters.

  19. The role of proteases, endoplasmic reticulum stress and SERPINA1 heterozygosity in lung disease and α-1 anti-trypsin deficiency.

    Science.gov (United States)

    Greene, Catherine M; Hassan, Tidi; Molloy, Kevin; McElvaney, Noel G

    2011-06-01

    The serine proteinase inhibitor α-1 anti-trypsin (AAT) provides an antiprotease protective screen throughout the body. Mutations in the AAT gene (SERPINA1) that lead to deficiency in AAT are associated with chronic obstructive pulmonary diseases. The Z mutation encodes a misfolded variant of AAT that is not secreted effectively and accumulates intracellularly in the endoplasmic reticulum of hepatocytes and other AAT-producing cells. Until recently, it was thought that loss of antiprotease function was the major cause of ZAAT-related lung disease. However, the contribution of gain-of-function effects is now being recognized. Here we describe how both loss- and gain-of-function effects can contribute to ZAAT-related lung disease. In addition, we explore how SERPINA1 heterozygosity could contribute to smoking-induced chronic obstructive pulmonary diseases and consider the consequences.

  20. The role of proteases, endoplasmic reticulum stress and SERPINA1 heterozygosity in lung disease and alpha-1 anti-trypsin deficiency.

    LENUS (Irish Health Repository)

    Greene, Catherine M

    2012-02-01

    The serine proteinase inhibitor alpha-1 anti-trypsin (AAT) provides an antiprotease protective screen throughout the body. Mutations in the AAT gene (SERPINA1) that lead to deficiency in AAT are associated with chronic obstructive pulmonary diseases. The Z mutation encodes a misfolded variant of AAT that is not secreted effectively and accumulates intracellularly in the endoplasmic reticulum of hepatocytes and other AAT-producing cells. Until recently, it was thought that loss of antiprotease function was the major cause of ZAAT-related lung disease. However, the contribution of gain-of-function effects is now being recognized. Here we describe how both loss- and gain-of-function effects can contribute to ZAAT-related lung disease. In addition, we explore how SERPINA1 heterozygosity could contribute to smoking-induced chronic obstructive pulmonary diseases and consider the consequences.

  1. Recognizing problem video game use.

    Science.gov (United States)

    Porter, Guy; Starcevic, Vladan; Berle, David; Fenech, Pauline

    2010-02-01

    It has been increasingly recognized that some people develop problem video game use, defined here as excessive use of video games resulting in various negative psychosocial and/or physical consequences. The main objectives of the present study were to identify individuals with problem video game use and compare them with those without problem video game use on several variables. An international, anonymous online survey was conducted, using a questionnaire with provisional criteria for problem video game use, which the authors have developed. These criteria reflect the crucial features of problem video game use: preoccupation with and loss of control over playing video games and multiple adverse consequences of this activity. A total of 1945 survey participants completed the survey. Respondents who were identified as problem video game users (n = 156, 8.0%) differed significantly from others (n = 1789) on variables that provided independent, preliminary validation of the provisional criteria for problem video game use. They played longer than planned and with greater frequency, and more often played even though they did not want to and despite believing that they should not do it. Problem video game users were more likely to play certain online role-playing games, found it easier to meet people online, had fewer friends in real life, and more often reported excessive caffeine consumption. People with problem video game use can be identified by means of a questionnaire and on the basis of the present provisional criteria, which require further validation. These findings have implications for recognition of problem video game users among individuals, especially adolescents, who present to mental health services. Mental health professionals need to acknowledge the public health significance of the multiple negative consequences of problem video game use.

  2. Protective effects of soybean protein and egg white protein on the antibacterial activity of nisin in the presence of trypsin.

    Science.gov (United States)

    Pan, Dan; Zhang, Dong; Hao, Limin; Lin, Songyang; Kang, Qiaozhen; Liu, Xin; Lu, Laizheng; Lu, Jike

    2018-01-15

    The using of nisin to prevent foodborne pathogens (Staphylococcus aureus and Listeria monocytogenes) from contamination has received broad attentions during meat processing. However, the application of nisin has been limited because its antibacterial activity may be inhibited by trypsin. In this study, the protective effects of soybean protein and egg white protein on antibacterial activity of nisin were evaluated. It could be concluded that exogenous trypsin decreased the antibacterial activity of nisin, soybean protein and egg white protein could keep the nisin activity from enzymolysis of trypsin. Trypsin inhibitors in soybean protein and egg white protein could protect the antibacterial activity of nisin. Nisin with soybean protein or egg white protein in cooked meat product presented better quality preservation effects than nisin alone in the presence of trypsin. The total viable counts (TVC) and total volatile basic nitrogen (TVB-N) of nisin-treated group were significantly higher than these in nisin-soybean protein-treated and nisin-egg white protein-treated groups with trypsin. This study showed the potential of using soybean protein and egg white protein to stabilize the antibacterial activity of nisin under high trypsin conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. A sensitive and real-time assay of trypsin by using molecular imprinting-based capacitive biosensor.

    Science.gov (United States)

    Ertürk, Gizem; Hedström, Martin; Mattiasson, Bo

    2016-12-15

    Use of a highly sensitive, selective capacitive biosensor is reported for label-free, real-time, easy and rapid detection of trypsin by using the microcontact imprinting method. Real-time trypsin detection was performed with trypsin-imprinted (trypsin-MIP) capacitive electrodes using standard trypsin solutions in the concentration range of 1.0×10(-13)-1.0×10(-7)M with a detection limit of 3.0×10(-13)M. Selectivity and cross-reactivity of the system were tested by using competing proteins including chymotrypsin (chy), bovine serum albumin (BSA), lysozyme (lyz) and cytochrome c (cyt c) in singular and competitive manner and the selectivity of the system was determined with the selectivity coefficients of approximately 705.1, 6.5, 6.4 and 5.1 for chy, BSA, lyz and cyt c, respectively. The trypsin-MIP capacitive electrode was used for ~80 assays during 2 months and retained its binding property during all that time with a decrease of approximately 2.3% in the signal amplitude. In the last step, trypsin activity was measured by using Nα-Benzoyl-D, l-arginine 4-nitroanilide hydrochloride (BAPNA) as the substrate with spectrophotometer at 410nm. The trypsin activity was measured as 9mU/mL by spectrophotometer while the amount of captured enzyme calculated from the capacitive system was 7.9mU/mL which shows the correlation between two methods. From the comparison it is obvious that the new method is an attractive alternative for assaying trypsin and the developed capacitive system might be used successfully to monitor label-free, real-time enzymatic activity of different proteases in a sensitive, rapid, cost-effective manner for different applications. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Dual core quantum dots for highly quantitative ratiometric detection of trypsin activity in cystic fibrosis patients

    Science.gov (United States)

    Castelló Serrano, Iván; Stoica, Georgiana; Matas Adams, Alba; Palomares, Emilio

    2014-10-01

    We present herein two colour encoded silica nanospheres (2nanoSi) for the fluorescence quantitative ratiometric determination of trypsin in humans. Current detection methods for cystic fibrosis diagnosis are slow, costly and suffer from false positives. The 2nanoSi proved to be a highly sensitive, fast (minutes), and single-step approach nanosensor for the screening and diagnosis of cystic fibrosis, allowing the quantification of trypsin concentrations in a wide range relevant for clinical applications (25-350 μg L-1). Furthermore, as trypsin is directly related to the development of cystic fibrosis (CF), different human genotypes, i.e. CF homozygotic, CF heterozygotic, and unaffected, respectively, can be determined using our 2nanoSi nanospheres. We anticipate the 2nanoSi system to be a starting point for non-invasive, easy-to-use and cost effective ratiometric fluorescent biomarkers for recessive genetic diseases like human cystic fibrosis. In a screening program in which the goal is to detect disease and also the carrier status, early diagnosis could be of great help.We present herein two colour encoded silica nanospheres (2nanoSi) for the fluorescence quantitative ratiometric determination of trypsin in humans. Current detection methods for cystic fibrosis diagnosis are slow, costly and suffer from false positives. The 2nanoSi proved to be a highly sensitive, fast (minutes), and single-step approach nanosensor for the screening and diagnosis of cystic fibrosis, allowing the quantification of trypsin concentrations in a wide range relevant for clinical applications (25-350 μg L-1). Furthermore, as trypsin is directly related to the development of cystic fibrosis (CF), different human genotypes, i.e. CF homozygotic, CF heterozygotic, and unaffected, respectively, can be determined using our 2nanoSi nanospheres. We anticipate the 2nanoSi system to be a starting point for non-invasive, easy-to-use and cost effective ratiometric fluorescent biomarkers for

  5. Vulvodynia; an under-recognized disease

    Directory of Open Access Journals (Sweden)

    Simona Roxana Georgescu

    2016-10-01

    Full Text Available Vulvodynia is a chronic condition which affects an increasing number of women; it presents currently an incidence that is higher than had previously been estimated. Regarding pathogenesis, several (hormonal, infectious, inflammatory and psychological factors have been proposed, but vulvodynia etiology remains still unclear. This disorder is a multifactorial condition with a significant impact on the patient’s quality of life, yet is difficult to diagnose (an under-estimated/ under-recognized affection. Certain medical investigations are required in order to exclude other diseases (the diagnosis of vulvodynia being one of exclusion, but anamnesis and physical examination are essential steps in the diagnosis. Although many therapies have been proposed, both pharmacological and nonpharmacological, a standardized therapy has not yet been established/ generally accepted. Accordingly, many therapeutic options have been studied with varying results. Vulvodynia remains a challenging disease and a multidisciplinary approach is needed to achieve satisfactory outcomes. Further studies are needed to completely understand its pathogenesis and to develop a standardized treatment.

  6. Overexpression of glycosylated proteins in cervical cancer recognized by the Machaerocereus eruca agglutinin.

    Science.gov (United States)

    Solórzano, Carlos; Angel Mayoral, Miguel; de los Angeles Carlos, María; Berumen, Jaime; Guevara, Jorge; Raúl Chávez, Francisco; Mendoza-Hernández, Guillermo; Agundis, Concepción; Zenteno, Edgar

    2012-10-08

    In cervical cancer, glycosylation has been suggested as being involved in both its carcinogenesis and invasive capacity. In this work, we analyzed mucin type O-glycosylation in biopsies of invasive cervical cancer in FIGO stage II B through histochemistry using lectins specific for O-glycosidically linked glycans. Our results reveal that the lectin Machaerocereus eruca (MeA, specific for Gal in a Fucα1,2 (GalNAcα1,3) Galβ1,4) showed increased recognition of tumoral cells and tumoral stroma tissue compared to other lectins with similar specificity; healthy cervical tissue was negative for MeA. Trypsin treatment of recognized tissues abolished MeA's recognition;moreover, interaction of MeA was inhibited with oligosaccharides from mucin. As demonstrated by Western blot of 2-D electrophoresis, MeA recognized ten glycoproteins in the range from 122 to 42 kDa in cervical cancer lysates. The LC-ESI-MS/MS analysis of the MeAs' recognized peptides revealed that the latter matched mainly with the amino acid sequences of lamin A/C, vimentin, elongation factor 2, keratin 1, and beta actin. Our results suggest that MeA recognizes a complex of over-expressed O-glycosidically-linked proteins that play a relevant role in cervical cancer's invasive capacity. O-glycosylation participates in the disassembly of intercellular junctions favoring cancer progression.

  7. Influence of heavy metal salts on the activity of trypsin-like hydrolases from Drosophila melanogaster

    Directory of Open Access Journals (Sweden)

    I. L. Ryzhko

    2014-10-01

    Full Text Available The influence of salts of heavy metals on trypsin-like peptide hydrolase of drosophila larvae partly refined by methods of salting-out, gel chromatography and electrophoresis has been researched. It is established that cadmium chloride is characterized by the greatest inhibitory effect, while zinc chloride by the lowest one. Since metal chlorides were used in all cases, it is the differentiated effect of metal ions on manifestations of amidase activity of trypsin-like peptide hydrolase of drosophila larvae, which rather may be considered as proved than the effect of chlorine ions. This, as a whole, agrees with the effect of these ions on proteolytic digestion system at the level of live organisms.

  8. Efficacy of Trypsin in Treating Coral Snake Envenomation in the Porcine Model.

    Science.gov (United States)

    Parker-Cote, Jennifer L; O'Rourke, Dorcas P; Brewer, Kori L; Lertpiriyapong, Kvin; Punja, Mohan; Bush, Sean P; Miller, Susan N; Meggs, William J

    2015-12-01

    Antivenom is the definitive treatment for venomous snakebites. Alternative treatments warrant investigation because antivenom is sometimes unavailable, expensive, and can have deleterious side effects. This study assesses the efficacy of trypsin to treat coral snake envenomation in an in vivo porcine model. A randomized, blinded study was conducted. Subjects were 13 pigs injected subcutaneously with 1 mL of eastern coral snake venom (10 mg/mL) in the right distal hind limb. After 1 min, subjects were randomized to have the envenomation site injected with either 1 mL of saline or 1 mL of trypsin (100 mg/mL) by a blinded investigator. Clinical endpoint was survival for 72 h or respiratory depression defined as respiratory rate coral snake venom and increased survival significantly. Further investigation is required before these results can be extended to human snakebites.

  9. Crystallization and preliminary X-ray diffraction studies of Murraya koenigii trypsin inhibitor

    Energy Technology Data Exchange (ETDEWEB)

    Shee, Chandan [Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247 667 (India); Singh, Tej P. [Department of Biophysics, All India Institute of Medical Sciences, New Delhi 100 029 (India); Kumar, Pravindra, E-mail: kumarfbs@iitr.ernet.in; Sharma, Ashwani K., E-mail: kumarfbs@iitr.ernet.in [Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247 667 (India)

    2007-04-01

    A Kunitz-type trypsin inhibitor purified from the seeds of Murraya koenigii has been crystallized by the sitting-drop vapour-diffusion method using PEG 8000 as the precipitating agent. A Kunitz-type trypsin inhibitor purified from the seeds of Murraya koenigii has been crystallized by the sitting-drop vapour-diffusion method using PEG 8000 as the precipitating agent. The crystals belong to the tetragonal space group P4{sub 3}2{sub 1}2, with unit-cell parameters a = b = 75.8, c = 150.9 Å. The crystals contain two molecules in the asymmetric unit with a V{sub M} value of 2.5 Å{sup 3} Da{sup −1}. Diffraction was observed to 2.65 Å resolution and a complete data set was collected to 2.9 Å resolution.

  10. Bauhinia variegata var. variegata trypsin inhibitor: From isolation to potential medicinal applications

    Energy Technology Data Exchange (ETDEWEB)

    Fang, Evandro Fei; Wong, Jack Ho [School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong SAR (China); Bah, Clara Shui Fern [Department of Food Science, Division of Sciences, University of Otago (New Zealand); Lin, Peng [School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong SAR (China); Tsao, Sai Wah [Department of Anatomy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Sassoon Road, Pokfulam, Hong Kong SAR (China); Ng, Tzi Bun, E-mail: b021770@mailserv.cuhk.edu.hk [School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong SAR (China)

    2010-06-11

    Here we report for the first time of a new Kunitz-type trypsin inhibitor (termed BvvTI) from seeds of the Camel's foot tree, Bauhinia variegata var. variegata. BvvTI shares the same reactive site residues (Arg, Ser) and exhibits a homology of N-terminal amino acid sequence to other Bauhinia protease inhibitors. The trypsin inhibitory activity (K{sub i}, 0.1 x 10{sup -9} M) of BvvTI ranks the highest among them. Besides anti-HIV-1 reverse transcriptase activity, BvvTI could significantly inhibit the proliferation of nasopharyngeal cancer CNE-1 cells in a selective way. This may partially be contributed by its induction of cytokines and apoptotic bodies. These results unveil potential medicinal applications of BvvTI.

  11. [Procedure for obtaining and catalytic properties of trypsin immobilized in cryogels of polyvinyl alcohol].

    Science.gov (United States)

    Lysogorskaia, E N; Rosliakova, T V; Beliaeva, A V; Bacheva, A V; Lozinskiĭ, V I; Filippova, I Iu

    2008-01-01

    Commercial preparations of trypsin, varying in activity, were immobilized in a cryogel of polyvinyl alcohol, activated by dialdehydes (terephthalic, succinic, or glutaric) or divinyl sulfone. All preparations of the immobilized enzyme exhibited hydrolytic activity and retained stability for 8 months. In an organic solvent environment, specimens of immobilized trypsin catalyzed the synthesis of N-carbobenzoxy-L-phenylalanyl-L-arginyl-L-leucine p-nitroanilide from N-carbobenzoxy-L-phenylalanyl-L-argininine methyl ester (or N-carbobenzoxy-L-phenylalanyl-L-arginine) and L-leucine p-nitroanilide, as well as the formation of N-carbobenzoxy-L-alanyl-L-alanyl-L-arginyl-L-phenylalanine p-nitroanilide from N-carbobenzoxy-L-alanyl-L-alanyl-L-arginine and L-phenylalanine p-nitroanilide. The presence of small amounts of water in organic solvents was prerequisite to the biocatalysts manifesting synthase activity in reactions of peptide bond formation.

  12. Why Ser and not Thr brokers catalysis in the trypsin fold.

    Science.gov (United States)

    Pelc, Leslie A; Chen, Zhiwei; Gohara, David W; Vogt, Austin D; Pozzi, Nicola; Di Cera, Enrico

    2015-02-24

    Although Thr is equally represented as Ser in the human genome and as a nucleophile is as good as Ser, it is never found in the active site of the large family of trypsin-like proteases that utilize the Asp/His/Ser triad. The molecular basis of the preference of Ser over Thr in the trypsin fold was investigated with X-ray structures of the thrombin mutant S195T free and bound to an irreversible active site inhibitor. In the free form, the methyl group of T195 is oriented toward the incoming substrate in a conformation seemingly incompatible with productive binding. In the bound form, the side chain of T195 is reoriented for efficient substrate acylation without causing steric clash within the active site. Rapid kinetics prove that this change is due to selection of an active conformation from a preexisting ensemble of reactive and unreactive rotamers whose relative distribution determines the level of activity of the protease. Consistent with these observations, the S195T substitution is associated with a weak yet finite activity that allows identification of an unanticipated important role for S195 as the end point of allosteric transduction in the trypsin fold. The S195T mutation abrogates the Na(+)-dependent enhancement of catalytic activity in thrombin, activated protein C, and factor Xa and significantly weakens the physiologically important allosteric effects of thrombomodulin on thrombin and of cofactor Va on factor Xa. The evolutionary selection of Ser over Thr in trypsin-like proteases was therefore driven by the need for high catalytic activity and efficient allosteric regulation.

  13. Targeting Trypsin-Inflammation Axis for Pancreatitis Therapy in a Humanized Pancreatitis Model

    Science.gov (United States)

    2016-10-01

    supraphysiological concentrations. CCK is a hormone that at physiological concentrations induces the release of digestive enzymes by pancreatic...death. We observed, that compared to WT cells, R122H cells display in basal conditions similar morphology and protein levels of digestive enzymes ...Award Number: W81XWH-15-1-0258 TITLE: Targeting Trypsin-Inflammation Axis for Pancreatitis Therapy in a Humanized Pancreatitis Model PRINCIPAL

  14. Bioinsecticidal activity of a novel Kunitz trypsin inhibitor from Catanduva (Piptadenia moniliformis) seeds.

    Science.gov (United States)

    Cruz, Ana C B; Massena, Fábio S; Migliolo, Ludovico; Macedo, Leonardo L P; Monteiro, Norberto K V; Oliveira, Adeliana S; Macedo, Francisco P; Uchoa, Adriana F; Grossi de Sá, Maria F; Vasconcelos, Ilka M; Murad, Andre M; Franco, Octavio L; Santos, Elizeu A

    2013-09-01

    The present study aims to provide new in vitro and in vivo biochemical information about a novel Kunitz trypsin inhibitor purified from Piptadenia moniliformis seeds. The purification process was performed using TCA precipitation, Trypsin-Sepharose and reversed-phase C18 HPLC chromatography. The inhibitor, named PmTKI, showed an apparent molecular mass of around 19 kDa, visualized by SDS-PAGE, which was confirmed by mass spectrometry MALDI-ToF demonstrating a monoisotopic mass of 19.296 Da. The inhibitor was in vitro active against trypsin, chymotrypsin and papain. Moreover, kinetic enzymatic studies were performed aiming to understand the inhibition mode of PmTKI, which competitively inhibits the target enzyme, presenting Ki values of 1.5 × 10(-8) and 3.0 × 10(-1) M against trypsin and chymotrypsin, respectively. Also, the inhibitory activity was assayed at different pH ranges, temperatures and reduction environments (DTT). The inhibitor was stable in all conditions maintaining an 80% residual activity. N-terminal sequence was obtained by Edman degradation and the primary sequence presented identity with members of Kunitz-type inhibitors from the same subfamily. Finally after biochemical characterization the inhibitory effect was evaluated in vitro on insect digestive enzymes from different orders, PmTKI demonstrated remarkable activity against enzymes from Anthonomus grandis (90%), Plodia interpuncptella (60%), and Ceratitis capitata (70%). Furthermore, in vivo bioinsecticidal assays of C. capitata larvae were also performed and the concentration of PmTKI (w/w) in an artificial diet required to LD50 and ED50 larvae were 0.37 and 0.3% respectively. In summary, data reported here shown the biotechnological potential of PmTKI for insect pest control. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  15. Immobilization of trypsin on miniature incandescent bulbs for infrared-assisted proteolysis

    Energy Technology Data Exchange (ETDEWEB)

    Ge, Huimin; Bao, Huimin; Zhang, Luyan; Chen, Gang, E-mail: gangchen@fudan.edu.cn

    2014-10-03

    Highlights: • Trypsin was immobilized on miniature incandescent bulbs via chitosan coating. • The bulbs acted as enzymatic reactors and the generators of infrared radiation. • The bulb bioreactors were successfully employed in infrared-assisted proteolysis. • The proteolysis could accomplish within 5 min with high sequence coverages. - Abstract: A novel efficient proteolysis approach was developed based on trypsin-immobilized miniature incandescent bulbs and infrared (IR) radiation. Trypsin was covalently immobilized in the chitosan coating on the outer surface of miniature incandescent bulbs with the aid of glutaraldehyde. When an illuminated enzyme-immobilized bulb was immersed in protein solution, the emitted IR radiation could trigger and accelerate heterogeneous protein digestion. The feasibility and performance of the novel proteolysis approach were demonstrated by the digestion of hemoglobin (HEM), cytochrome c (Cyt-c), lysozyme (LYS), and ovalbumin (OVA) and the digestion time was significantly reduced to 5 min. The obtained digests were identified by MALDI-TOF-MS with the sequence coverages of 91%, 77%, 80%, and 52% for HEM, Cyt-c, LYS, and OVA (200 ng μL{sup −1} each), respectively. The suitability of the prepared bulb bioreactors to complex proteins was demonstrated by digesting human serum.

  16. Crystal structure of an engineered subtilisin inhibitor complexed with bovine trypsin.

    Science.gov (United States)

    Takeuchi, Y; Nonaka, T; Nakamura, K T; Kojima, S; Miura, K; Mitsui, Y

    1992-05-15

    Proteinase specificity of a proteinaceous inhibitor of subtilisin (SSI; Streptomyces subtilisin inhibitor) can be altered so as to strongly inhibit trypsin simply by replacing P1 methionine with lysine (with or without concomitant change of the P4 residue) through site-directed mutagenesis. Now the crystal structure of one such engineered SSI (P1 methionine converted to lysine and P4 methionine converted to glycine) complexed with bovine trypsin has been solved at 2.6 A resolution and refined to a crystallographic R factor of 0.173. Comparing this structure with the previously established structure of the native SSI complexed with subtilisin BPN', it was found that (i) P1 lysine of the mutant SSI is accommodated in the S1 pocket of trypsin as usual, and (ii) upon complex formation, considerable conformation change occurs to the reactive site loop of the mutant SSI. Thus, in this case, flexibility of the reactive site loop seems important for successfully changing the proteinase specificity through mere replacement of the P1 residue.

  17. Structural characterization of Spinacia oleracea trypsin inhibitor III (SOTI-III).

    Science.gov (United States)

    Glotzbach, Bernhard; Schmelz, Stefan; Reinwarth, Michael; Christmann, Andreas; Heinz, Dirk W; Kolmar, Harald

    2013-01-01

    In recent decades, several canonical serine protease inhibitor families have been classified and characterized. In contrast to most trypsin inhibitors, those from garden four o'clock (Mirabilis jalapa) and spinach (Spinacia oleracea) do not share sequence similarity and have been proposed to form the new Mirabilis serine protease inhibitor family. These 30-40-amino-acid inhibitors possess a defined disulfide-bridge topology and belong to the cystine-knot miniproteins (knottins). To date, no atomic structure of this inhibitor family has been solved. Here, the first structure of S. oleracea trypsin inhibitor III (SOTI-III), in complex with bovine pancreatic trypsin, is reported. The inhibitor was synthesized by solid-phase peptide synthesis on a multi-milligram scale and was assayed to test its inhibitory activity and binding properties. The structure confirmed the proposed cystine-bridge topology. The structural features of SOTI-III suggest that it belongs to a new canonical serine protease inhibitor family with promising properties for use in protein-engineering and medical applications.

  18. Structural and Chemical Characterization of Silica Spheres before and after Modification by Silanization for Trypsin Immobilization

    Directory of Open Access Journals (Sweden)

    Eduardo F. Barbosa

    2017-01-01

    Full Text Available In the last decades, silica particles of a variety of sizes and shapes have been characterized and chemically modified for several applications, from chromatographic separation to dental supplies. The present study proposes the use of aminopropyl triethoxysilane (APTS silanized silica particles to immobilize the proteolytic enzyme trypsin for the development of a bioreactor. The major advantage of the process is that it enables the polypeptides hydrolysis interruption simply by removing the silica particles from the reaction bottle. Silanized silica surfaces showed significant morphological changes at micro- and nanoscale level. Chemical characterization showed changes in elemental composition, chemical environment, and thermal degradation. Their application as supports for trypsin immobilization showed high immobilization efficiency at reduced immobilization times, combined with more acidic conditions. Indirect immobilization quantification by reversed-phase ultrafast high performance liquid chromatography proved to be a suitable approach due to its high linearity and sensitivity. Immobilized trypsin activities on nonmodified and silanized silica showed promising features (e.g., selective hydrolysis for applications in proteins/peptides primary structure elucidation for proteomics. Silanized silica system produced some preferential targeting peptides, probably due to the hydrophobicity of the nanoenvironment conditioned by silanization.

  19. Extraction, purification and characterization of inhibitor of trypsin from Chenopodium quinoa seeds

    Directory of Open Access Journals (Sweden)

    Aline Regiele Pesoti

    2015-01-01

    Full Text Available AbstractA novel trypsin inhibitor of protease (CqTI was purified from Chenopodium quinoa seeds. The optimal extracting solvent was 0.1M NaCl pH 6.8 (p < 0.05. The extraction time of 5h and 90 °C was optimum for the recovery of the trypsin inhibitor from C. quinoa seeds. The purification occurred in gel-filtration and reverse phase chromatography. CqTI presented active against commercial bovine trypsin and chymotrypsin and had a specific activity of 5,033.00 (TIU/mg, which was purified to 333.5-fold. The extent of purification was determined by SDS-PAGE. CqTI had an apparent molecular weight of approximately 12KDa and two bands in reduced conditions as determined by Tricine-SDS-PAGE. MALDI-TOF showed two peaks in 4,246.5 and 7,908.18m/z. CqTI presented high levels of essential amino acids. N-terminal amino acid sequence of this protein did not show similarity to any known protease inhibitor. Its activity was stable over a pH range (2-12, temperatures range (20-100 °C and reducing agents.

  20. Preparation and characterization of magnetic levan particles as matrix for trypsin immobilization

    Energy Technology Data Exchange (ETDEWEB)

    Maciel, J.C. [Programa de Pos-Graduacao em Ciencias Biologicas, Universidade Federal de Pernambuco, Cidade Universitaria, 50670-901 Recife, PE (Brazil); Andrad, P.L. [Programa de Pos-Graduacao em Ciencia de Materiais, Universidade Federal de Pernambuco, Cidade Universitaria, 50679-901 Recife, PE (Brazil); Neri, D.F.M., E-mail: davidfmneri@yahoo.com.br [Universidade Federal do Vale do Sao Francisco, 56304-205 Petrolina, PE (Brazil); Carvalho, L.B. [Departamento de Bioquimica, Universidade Federal de Pernambuco, Cidade Universitaria, 50679-901 Recife, PE (Brazil); Cardoso, C.A. [Departamento de Fisica, Universidade Federal de Sao Carlos, 13565-905 Sao Carlos, PE (Brazil); Calazans, G.M.T. [Departamento de Antibioticos, Universidade Federal de Pernambuco, Cidade Universitaria, 50670-901 Recife, PE (Brazil); Albino Aguiar, J. [Departamento de Fisica, Universidade Federal de Pernambuco, Cidade Universitaria, 50679-901 Recife, PE (Brazil); Silva, M.P.C. [Departamento de Bioquimica, Universidade Federal de Pernambuco, Cidade Universitaria, 50679-901 Recife, PE (Brazil)

    2012-04-15

    Magnetic levan was synthesized by co-precipitating D-fructofuranosyl homopolysaccharide with a solution containing Fe{sup 2+} and Fe{sup 3+} in alkaline conditions at 100 Degree-Sign C. The magnetic levan particles were characterized by scanning electron microscopy (SEM), magnetization measurements, X-ray diffractometry (XRD) and infrared spectroscopy (IR). Afterwards, magnetic levan particles were functionalized by NaIO{sub 4} oxidation and used as matrices for trypsin covalent immobilization. Magnetite and magnetic levan particles were both heterogeneous in shape and levan-magnetite presented bigger sizes compared to magnetite according to SEM images. Magnetic levan particles exhibited a magnetization 10 times lower as compared to magnetite ones, probably, due to the coating layer. XRD diffractogram showed that magnetite is the dominant phase in the magnetic levan. Infrared spectroscopy showed characteristics absorption bands of levan and magnetite (O-H, C-O-C and Fe-O bonds). The immobilized trypsin derivative was reused 10 times and lost 16% of its initial specific activity only. Therefore, these magnetic levan particles can be proposed as an alternative matrices for enzyme immobilization. - Highlights: Black-Right-Pointing-Pointer The magnetic levan particles presented larger size variation than magnetite particles due to the changes produced by coating. Black-Right-Pointing-Pointer The utilization of magnetic levan particles showed to be efficacious for immobilization of enzymes as trypsin. Black-Right-Pointing-Pointer Magnetic particles can be planned as other matrix for immobilization of biomolecule in various division processes in biotechnology.

  1. Influence of carbohydrates on the interaction of procyanidin B3 with trypsin.

    Science.gov (United States)

    Gonçalves, Rui; Mateus, Nuno; De Freitas, Victor

    2011-11-09

    The biological properties of procyanidins, in particular their inhibition of digestive enzymes, have received much attention in the past few years. Dietary carbohydrates are an environmental factor that is known to affect the interaction of procyanidins with proteins. This work aimed at understanding the effect of ionic food carbohydrates (polygalacturonic acid, arabic gum, pectin, and xanthan gum) on the interaction between procyanidins and trypsin. Physical-chemical techniques such as saturation transfer difference-NMR (STD-NMR) spectroscopy, fluorescence quenching, and nephelometry were used to evaluate the interaction process. Using STD-NMR, it was possible to identify the binding of procyanidin B3 to trypsin. The tested carbohydrates prevented the association of procyanidin B3 and trypsin by a competition mechanism in which the ionic character of carbohydrates and their ability to encapsulate procyanidins seem crucial leading to a reduction in STD signal and light scattering and to a recovery of the proteins intrinsic fluorescence. On the basis of these results, it was possible to grade the carbohydrates in their aggregation inhibition ability: XG > PA > AG ≫ PC. These effects may be relevant since the coingestion of procyanidins and ionic carbohydrates are frequent and furthermore since these might negatively affect the antinutritional properties ascribed to procyanidins in the past.

  2. Inhibition of Amyloid-like Fibril Formation of Trypsin by Red Wines.

    Science.gov (United States)

    Kotorman, Marta; Kasi, Phanindra B; Halasz, Laszlo; Borics, Attila

    2017-01-01

    The aim of the present study was to examine the potential role and applicability of dietary supplements in reducing the risk of development of amyloid diseases associated with the gastrointestinal tract, such as type II diabetes. Trypsin, a well-known serine protease was used as a model protein in our experiments. The effect of various red wines on the formation of amyloid-like fibrils of trypsin was studied in vitro, in aqueous ethanol, at pH 7.0. Turbidity measurements, aggregation kinetics experiments, Congo red binding assays and electronic circular dichroism spectroscopic measurements were used to follow the aggregation process in the presence or absence of various red wines. The results suggest that red wines effectively inhibit the formation of amyloid-like fibrils of trypsin and the inhibitory effect is dose-dependent. The extent of inhibition was found to be proportional to the total concentration of phenolic compounds. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  3. Trypsin inhibition activity of heat-denatured ovomucoid: a kinetic study.

    Science.gov (United States)

    Van der Plancken, Iesel; Van Remoortere, Marijke; Van Loey, Ann; Hendrickx, Marc E

    2004-01-01

    A kinetic study was conducted on the effect of heating in the temperature range of 75-110 degrees C on the trypsin inhibition activity of ovomucoid. Heat treatment of isolated ovomucoid resulted in a time-dependent decrease in trypsin inhibition activity that could accurately be described by a first-order kinetic model. The magnitude and the temperature dependence of the rate constants was affected by the pH during heat treatment. The heat stability of ovomucoid was the lowest at pH 7.6. Heat treatments intended to decrease the trypsin inhibition activity should therefore be carried out as soon as possible after laying, because the ovomucoid was inactivated faster at the pH of fresh egg white (pH 7.6). The presence of the other egg white constituents decreased the heat stability of ovomucoid compared to that of the model system of ovomucoid in buffer, presumably by the formation of ovomucoid-lysozyme complexes in the former.

  4. Allergenicity, trypsin inhibitor activity and nutritive quality of enzymatically modified soy proteins.

    Science.gov (United States)

    De La Barca, Ana María Calderón; Wall, Abraham; López-Díaz, José Alberto

    2005-05-01

    Two ultrafiltered soy flour protein fractions were evaluated; the first was obtained by hydrolysis (0.5-3 kDa, F(0.5-3)), and the second was an enzymatically methionine-enriched fraction (1-10 kDa, F(1-10)E). Amino acid profiles, protein quality, allergenicity (against soy-sensitive infant sera) and trypsin inhibitor activity were determined. Fraction F(1-10)E fulfilled amino acid requirements for infants, whereas the F(0.5-3) fraction was methionine deficient. Both fractions were similar in net protein utilization, and F(1-10)E digestibility was comparable with casein and higher (P?soy isolate. Allergenicity of SF was reduced to 21.5% with the hydrolysis in F(1-10)E and it was not detected in F(0.5-3.) Residual trypsin inhibitor activity with respect to soy flour was 8.1%, 3.3% and 1% for hydrolysate, F(1-10)E and F(0.5-3), respectively. Both fractions presented high nutritive quality and reduced or null allergenicity. The trypsin inhibitor activity decreased along processing and could be a useful indicator for production of hypoallergenic proteins.

  5. Amino Acid Composition, Urease Activity and Trypsin Inhibitor Activity after Toasting of Soybean in Thick and Thin Layer

    OpenAIRE

    Tajana Krička; Vanja Jurišić; Neven Voća; Duška Ćurić; Tea Brlek Savić; Ana Matin

    2009-01-01

    The objective of this study was to determine amino acid content, urease activity and trypsin inhibitor activity in soybean grain for polygastric animals’ feed aft er toasting with the aim to introduce thick layer in toasting technology. Hence, soybean was toasted both in thick and thin layer at 130 oC during 10 minutes. In order to properly monitor the technological process of soybean thermal processing, it was necessary to study crude protein content, urease activity, trypsin inhibitor activ...

  6. Label-Free Fluorescent Detection of Trypsin Activity Based on DNA-Stabilized Silver Nanocluster-Peptide Conjugates

    Directory of Open Access Journals (Sweden)

    Cai-Xia Zhuo

    2016-11-01

    Full Text Available Trypsin is important during the regulation of pancreatic exocrine function. The detection of trypsin activity is currently limited because of the need for the substrate to be labeled with a fluorescent tag. A label-free fluorescent method has been developed to monitor trypsin activity. The designed peptide probe consists of six arginine molecules and a cysteine terminus and can be conjugated to DNA-stabilized silver nanoclusters (DNA-AgNCs by Ag-S bonding to enhance fluorescence. The peptide probe can also be adsorbed to the surface of graphene oxide (GO, thus resulting in the fluorescence quenching of DNA-AgNCs-peptide conjugate because of Förster resonance energy transfer. Once trypsin had degraded the peptide probe into amino acid residues, the DNA-AgNCs were released from the surface of GO, and the enhanced fluorescence of DNA-AgNCs was restored. Trypsin can be determined with a linear range of 0.0–50.0 ng/mL with a concentration as low as 1 ng/mL. This label-free method is simple and sensitive and has been successfully used for the determination of trypsin in serum. The method can also be modified to detect other proteases.

  7. Overexpression of glycosylated proteins in cervical cancer recognized by the Machaerocereus eruca agglutinin Overexpression of glycosylated proteins in cervical cancer recognized by the Machaerocereus eruca agglutinin

    Directory of Open Access Journals (Sweden)

    Carlos Solórzano

    2012-10-01

    Full Text Available In cervical cancer, glycosylation has been suggested as being involved in both its carcinogenesis and
    invasive capacity. In this work, we analyzed mucin type O-glycosylation in biopsies of invasive cervical cancer in
    FIGO stage II B through histochemistry using lectins specific for O-glycosidically linked glycans. Our results
    reveal that the lectin Machaerocereus eruca (MeA, specific for Gal in a Fuca1,2 (GalNAca1,3 Galb1,4 showed
    increased recognition of tumoral cells and tumoral stroma tissue compared to other lectins with similar specificity;
    healthy cervical tissue was negative for MeA. Trypsin treatment of recognized tissues abolished MeA’s recognition;
    moreover, interaction of MeA was inhibited with oligosaccharides from mucin. As demonstrated by
    Western blot of 2-D electrophoresis, MeA recognized ten glycoproteins in the range from 122 to 42 kDa in
    cervical cancer lysates. The LC-ESI-MS/MS analysis of the MeAs’ recognized peptides revealed that the latter
    matched mainly with the amino acid sequences of lamin A/C, vimentin, elongation factor 2, keratin 1, and beta
    actin. Our results suggest that MeA recognizes a complex of over-expressed O-glycosidically-linked proteins that
    play a relevant role in cervical cancer’s invasive capacity. O-glycosylation participates in the disassembly of intercellular
    junctions favoring cancer progression.In cervical cancer, glycosylation has been suggested as being involved in both its carcinogenesis and
    invasive capacity. In this work, we analyzed mucin type O-glycosylation in biopsies of invasive cervical cancer in
    FIGO stage II B through histochemistry using lectins specific for O-glycosidically linked glycans. Our results
    reveal that the lectin Machaerocereus eruca (MeA, specific for Gal in a Fuca1,2 (GalNAca1,3 Galb1,4 showed
    increased recognition of tumoral cells and tumoral stroma tissue compared to other lectins

  8. TRAFFIC SIGN RECOGNATION WITH VIDEO PROCESSING TECHNIQUE

    Directory of Open Access Journals (Sweden)

    Musa AYDIN

    2013-01-01

    Full Text Available In this study, traffic signs are aimed to be recognized and identified from a video image which is taken through a video camera. To accomplish our aim, a traffic sign recognition program has been developed in MATLAB/Simulink environment. The target traffic sign are recognized in the video image with the developed program.

  9. Teaching Students to Recognize Irony

    Science.gov (United States)

    Milner, Joseph O.; Hawkins, Robin H.; Milner, Lucy M.

    2014-01-01

    This article exposes the problem of using declarative rather than procedural knowledge to help K--12 students recognize irony in stories. It offers commonplace procedures drawn from students' everyday language experience together with more abstract irony clues to help students recognize irony in stories and increase their story comprehension.…

  10. The on-bead digestion of protein corona on nanoparticles by trypsin immobilized on the magnetic nanoparticle.

    Science.gov (United States)

    Hu, Zhengyan; Zhao, Liang; Zhang, Hongyan; Zhang, Yi; Wu, Ren'an; Zou, Hanfa

    2014-03-21

    Proteins interacting with nanoparticles would form the protein coronas on the surface of nanoparticles in biological systems, which would critically impact the biological identities of nanoparticles and/or result in the physiological and pathological consequences. The enzymatic digestion of protein corona was the primary step to achieve the identification of protein components of the protein corona for the bottom-up proteomic approaches. In this study, the investigation on the tryptic digestion of protein corona by the immobilized trypsin on a magnetic nanoparticle was carried out for the first time. As a comparison with the usual overnight long-time digestion and the severe self-digestion of free trypsin, the on-bead digestion of protein corona by the immobilized trypsin could be accomplished within 1h, along with the significantly reduced self-digestion of trypsin and the improved reproducibility on the identification of proteins by the mass spectrometry-based proteomic approach. It showed that the number of identified bovine serum (BS) proteins on the commercial Fe3O4 nanoparticles was increased by 13% for the immobilized trypsin with 1h digestion as compared to that of using free trypsin with even overnight digestion. In addition, the on-bead digestion of using the immobilized trypsin was further applied on the identification of human plasma protein corona on the commercial Fe3O4 nanoparticles, which leads the efficient digestion of the human plasma proteins and the identification of 149 human plasma proteins corresponding to putative critical pathways and biological processes. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Quantitative structure-activity relationships for organophosphates binding to trypsin and chymotrypsin.

    Science.gov (United States)

    Ruark, Christopher D; Hack, C Eric; Robinson, Peter J; Gearhart, Jeffery M

    2011-01-01

    Organophosphate (OP) nerve agents such as sarin, soman, tabun, and O-ethyl S-[2-(diisopropylamino) ethyl] methylphosphonothioate (VX) do not react solely with acetylcholinesterase (AChE). Evidence suggests that cholinergic-independent pathways over a wide range are also targeted, including serine proteases. These proteases comprise nearly one-third of all known proteases and play major roles in synaptic plasticity, learning, memory, neuroprotection, wound healing, cell signaling, inflammation, blood coagulation, and protein processing. Inhibition of these proteases by OP was found to exert a wide range of noncholinergic effects depending on the type of OP, the dose, and the duration of exposure. Consequently, in order to understand these differences, in silico biologically based dose-response and quantitative structure-activity relationship (QSAR) methodologies need to be integrated. Here, QSAR were used to predict OP bimolecular rate constants for trypsin and α-chymotrypsin. A heuristic regression of over 500 topological/constitutional, geometric, thermodynamic, electrostatic, and quantum mechanical descriptors, using the software Ampac 8.0 and Codessa 2.51 (SemiChem, Inc., Shawnee, KS), was developed to obtain statistically verified equations for the models. General models, using all data subsets, resulted in R(2) values of .94 and .92 and leave-one-out Q(2) values of 0.9 and 0.87 for trypsin and α-chymotrypsin. To validate the general model, training sets were split into independent subsets for test set evaluation. A y-randomization procedure, used to estimate chance correlation, was performed 10,000 times, resulting in mean R(2) values of .24 and .3 for trypsin and α-chymotrypsin. The results show that these models are highly predictive and capable of delineating the complex mechanism of action between OP and serine proteases, and ultimately, by applying this approach to other OP enzyme reactions such as AChE, facilitate the development of biologically based

  12. Some biochemical properties of trypsin inhibitor type antinutrients derived from extracts of wheat grain, Beta variety.

    Science.gov (United States)

    Mossor, G; Skupin, J

    1985-01-01

    Trypsin inhibitors were isolated and partially purified from wheat grain, Beta variety. The procedure for determination of the inhibitory activity was simplified. This pertains shortening of the reaction time as well as quantitative decrease of components in the incubation mixture. The inhibitory fraction was salted out at 30-65% ammonium sulfate saturation. The experimental material has not been initially defatted. For isolation of the inhibitors pH 4.4 was demonstrated to be optimal. The trypsin inhibitor was characterized by relatively low activity against trypsin (1.5% of soya inhibitory activity). Preparations showing inhibitory properties when stored at -18 degrees C retained their original activity for 40 days, whereas at 4 degrees C only for 10 days, respectively. Storage at 18 degrees C for 10 days resulted in 50% loss of the original activity. Among various factors stabilizing the inhibitory activity being studied, 2-mercaptoethanol at 0.01% final concentration was found to be most effective. Study on the effect of temperature on the antitrypsin activity revealed that the preparation retained its initial activity up to 80 degrees C. It has been demonstrated by wheat proteins fractionation that both albumin and globulin fractions were accompanied by the antitrypsin activity. Moreover gluten was also shown to exhibit some inhibitory activity. Variations in the inhibitory activity were evidenced during germination of wheat grain. After 2 days period of germination it tended to decrease, disappearing completely at the fifth day, respectively. The inhibitory activity appeared in coleoptile and root at fourth day of germination, being higher in coleoptile than in the roots.

  13. A Kunitz trypsin inhibitor of Entada scandens seeds: another member with single disulfide bridge.

    Science.gov (United States)

    Lingaraju, M H; Gowda, Lalitha R

    2008-05-01

    Sword bean (Entada scandens) is a tree climber that belongs to Mimosoideae, a subfamily of Leguminosae. A potent Kunitz type trypsin inhibitor (ESTI) was purified to homogeneity from Entada scandens seeds by sequential ammonium sulfate precipitation, affinity chromatography on trypsin-Sepharose and DEAE-Sephacel ion-exchange chromatography. ESTI is a single polypeptide chain of 19,766 Da. Both native PAGE as well as isoelectric focusing showed a single inhibitor species with a pI of 7.43. MALDI-TOF analysis also confirmed the monomeric nature. The amino-terminal sequence of ESTI reveals significant homology to the Kunitz-type protease inhibitors of legume plants. ESTI is unique in that it contains a single disulfide bridge, and unlike other inhibitors from Mimosoideae species is a single chain polypeptide. ESTI inhibited bovine trypsin with a stoichiometry of 1:1 and the apparent K(i) was 4.9 x 10(-9) M. In vitro assay showed that ESTI inhibited the midgut proteinase of the fifth instar larvae of Rice moth (Corcyra cephalonica) with an IC(50) of 26.4+/-0.01 nM. ESTI exhibits a mixed type competitive inhibition at lower concentration and pure competitive at higher inhibitor concentrations. Phylogenetic analyses depicted a clear divergence of single disulfide containing inhibitors from other tree legume Kunitz inhibitors. The homology of ESTI to Kunitz inhibitors together with the absence of Bowman-Birk type inhibitors in sword bean further supports the theory that there exists an evolutionary relationship between the families of inhibitors found in Leguminosae.

  14. Trypsin as enhancement in cyclical tracheal decellularization: Morphological and biophysical characterization

    Energy Technology Data Exchange (ETDEWEB)

    Giraldo-Gomez, D.M., E-mail: davidmauro2008@gmail.com [Posgrado en Ciencia e Ingeniería de Materiales, Universidad Nacional Autónoma de México (UNAM), Unidad de Posgrado Edificio “C” 1er Piso, Circuito de Posgrados, Avenida Universidad 3000, Ciudad Universitaria, Coyoacán, C.P. 04510, México D. F., México (Mexico); Instituto de Investigaciones en Materiales, Universidad Nacional Autónoma de México (UNAM), Circuito Exterior, Avenida Universidad 3000, Ciudad Universitaria, Coyoacán, C.P. 04510, México D.F., México (Mexico); Leon-Mancilla, B. [Departamento de Cirugía, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), Edificio “D” Planta Baja, Circuito Interior, Avenida Universidad 3000, Ciudad Universitaria, Coyoacán, C.P. 04510, México D.F., México (Mexico); Del Prado-Audelo, M.L. [Instituto de Investigaciones en Materiales, Universidad Nacional Autónoma de México (UNAM), Circuito Exterior, Avenida Universidad 3000, Ciudad Universitaria, Coyoacán, C.P. 04510, México D.F., México (Mexico); and others

    2016-02-01

    There are different types of tracheal disorders (e.g. cancer, stenosis and fractures). These can cause respiratory failure and lead to death of patients. Several attempts have been made for trachea replacement in order to restore the airway, including anastomosis and implants made from synthetic or natural materials. Tracheal allotransplantation has shown high rejection rates, and decellularization has emerged as a possible solution. Decellularization involves the removal of antigens from cells in the organ or tissue, leaving a matrix that can be used as 3D cell-scaffold. Although this process has been used for tracheal replacement, it usually takes at least two months and time is critical for patients with tracheal disorders. Therefore, there is necessary to develop a tracheal replacement process, which is not only effective, but also quick to prepare. The aim of this research was to develop a faster trachea decellularization protocol using Trypsin enzyme and Ethylenediaminetetraacetic acid (EDTA) as decellularization agents. Three protocols of cyclic trachea decellularization (Protocols A, B, and C) were compared. Following Protocol A (previously described in the literature), 15 consecutive cycles were performed over 32 days. Protocol B (a variation of Protocol A) — EDTA being added — with 15 consecutive cycles performed over 60 days. Finally, Protocol C, with the addition of Trypsin as a decellularization agent, 5 consecutive cycles being performed over 10 days. For the three protocols, hematoxylin–eosin (H&E) staining and DNA residual content quantification were performed to establish the effectiveness of the decellularization process. Scanning Electron Microscopy (SEM) was used to observe the changes in porosity and microarrays. To evaluate the structural matrices integrity, Thermogravimetric Analysis (TGA) and biomechanical test were used. None of the protocols showed significant alteration or degradation in the components of the extracellular matrix

  15. Crystallization and preliminary X-ray diffraction studies of Murraya koenigii trypsin inhibitor

    Science.gov (United States)

    Shee, Chandan; Singh, Tej P.; Kumar, Pravindra; Sharma, Ashwani K.

    2007-01-01

    A Kunitz-type trypsin inhibitor purified from the seeds of Murraya koenigii has been crystallized by the sitting-drop vapour-diffusion method using PEG 8000 as the precipitating agent. The crystals belong to the tetragonal space group P43212, with unit-cell parameters a = b = 75.8, c = 150.9 Å. The crystals contain two molecules in the asymmetric unit with a V M value of 2.5 Å3 Da−1. Diffraction was observed to 2.65 Å resolution and a complete data set was collected to 2.9 Å resolution. PMID:17401205

  16. Effects of gamma radiation on total phenolics, trypsin and tannin inhibitors in soybean grains

    Energy Technology Data Exchange (ETDEWEB)

    de Toledo, T.C.F [Department of Agroindustry, Food and Nutrition Escola Superior de Agricultura ' Luiz de Queiroz' , Universidade de Sao Paulo (ESALQ/USP), Piracicaba, SP (Brazil)]. E-mail: tcftoled@esalq.usp.br; Canniatti-Brazaca, S.G. [Department of Agroindustry, Food and Nutrition, Escola Superior de Agricultura ' Luiz de Queiroz' , Universidade de Sao Paulo (ESALQ/USP), Av. Padua Dias, 11 Box 9, CEP 13418-900, Piracicaba, SP (Brazil)]. E-mail: sgcbraza@esalq.usp.br; Arthur, V. [Food Irradiation and Entomology Laboratory, Center for Nuclear Agriculture-CENA/USP, Av. Centenario 303, Caixa Postal 96, 13400-970, Piracicaba, SP (Brazil)]. E-mail: arthur@cena.usp.br; Piedade, S.M.S. [Department of Mathematic Sciences, Escola Superior de Agricultura ' Luiz de Queiroz' , Universidade de Sao Paulo (ESALQ/USP), Piracicaba, SP (Brazil)]. E-mail: soniamsp@esalq.usp.br

    2007-10-15

    The objective was determining possible radiation-induced alterations (with doses of 2, 4 and 8 kGy) in raw or cooked grains from five soybean cultivars through the analysis of some antinutrient. Total phenolic ranged from 2.46 to 10.83 mg/g, the trypsin inhibited from 18.19 to 71.64 UTI/g and tannins from 0.01 to 0.39 mg/g. All the antinutrient studied underwent reduction with increases in the doses and cooking process was effective too.

  17. Characterization of ß-trypsin at acid pH by differential scanning calorimetry

    Directory of Open Access Journals (Sweden)

    Bittar E.R.

    2003-01-01

    Full Text Available Trypsin is a serino-protease with a polypeptide chain of 223 amino acid residues and contains six disulfide bridges. It is a globular protein with a predominance of antiparallel ß-sheet and helix in its secondary structure and has two domains with similar structures. We assessed the stability of ß-trypsin in the acid pH range using microcalorimetric (differential scanning calorimetry techniques. Protein concentrations varied in the range of 0.05 to 2.30 mg/ml. Buffer solutions of 50.0 mM ß-alanine and 20.0 mM CaCl2 at different pH values (from 2.0 to 4.2 and concentrations of sorbitol (1.0 and 2.0 M, urea (0.5 M or guanidinium hydrochloride (0.5 and 1.0 M were used. The data suggest that we are studying the same conformational transition of the protein in all experimental situations using pH, sorbitol, urea and guanidinium hydrochloride as perturbing agents. The observed van't Hoff ratios (deltaHcal/deltaHvH of 1.0 to 0.5 in the pH range of 3.2 to 4.2 suggest protein aggregation. In contrast, deltaHcal/deltaHvH ratios equal to one in the pH range of 2.0 to 3.2 suggest that the protein unfolds as a monomer. At pH 3.00, ß-trypsin unfolded with Tm = 54ºC and deltaH = 101.8 kcal/mol, and the change in heat capacity between the native and unfolded forms of the protein (deltaCp was estimated to be 2.50 ± 0.07 kcal mol-1 K-1. The stability of ß-trypsin calculated at 298 K was deltaG D = 5.7 kcal/mol at pH 3.00 and deltaG D = 15.2 kcal/mol at pH 7.00, values in the range expected for a small globular protein.

  18. A novel poly(deep eutectic solvent)-based magnetic silica composite for solid-phase extraction of trypsin

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Kaijia [State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082 (China); Wang, Yuzhi, E-mail: wyzss@hnu.edu.cn [State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082 (China); Li, Yixue; Lin, Yunxuan; Zhang, Haibao [State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082 (China); Zhou, Yigang [Department of Microbiology, College of Basic Medicine, Central South University, Changsha, 410083 (China)

    2016-11-23

    Novel poly(deep eutectic solvent) grafted silica-coated magnetic microspheres (Fe{sub 3}O{sub 4}@SiO{sub 2}-MPS@PDES) were prepared by polymerization of choline chloride-itaconic acid (ChCl-IA) and γ-MPS-modified magnetic silica composites, and were characterized by vibrating sample magnetometer (VSM), Fourier transform infrared spectrometry (FT-IR), X-ray photoelectron spectra (XPS), thermal gravimetric analysis (TGA) and transmission electron microscope (TEM). Then the synthetic Fe{sub 3}O{sub 4}@SiO{sub 2}-MPS@PDES microspheres were applied for the magnetic solid-phase extraction (MSPE) of trypsin for the first time. After extraction, the concentration of trypsin in the supernatant was determined by a UV–vis spectrophotometer. Single factor experiments were carried out to investigate the effects of the extraction process, including the concentration of trypsin, the ionic strength, the pH value, the extraction time and the temperature. Experimental results showed the extraction capacity could reach up to 287.5 mg/g under optimized conditions. In comparison with Fe{sub 3}O{sub 4}@SiO{sub 2}-MPS, Fe{sub 3}O{sub 4}@SiO{sub 2}-MPS@PDES displayed higher extraction capacity and selectivity for trypsin. According to the regeneration studies, Fe{sub 3}O{sub 4}@SiO{sub 2}-MPS@PDES microspheres can be recycled six times without significant loss of its extraction capacity, and retained a high extraction capacity of 233 mg/g after eight cycles. Besides, the activity studies also demonstrated that the activity of the extracted trypsin was well retained. Furthermore, the analysis of real sample revealed that the prepared magnetic microspheres can be used to purify trypsin in crude bovine pancreas extract. These results highlight the potential of the proposed Fe{sub 3}O{sub 4}@SiO{sub 2}-MPS@PDES-MSPE method in separation of biomolecules. - Highlights: • A strategy for solid-phase extraction of trypsin based on poly(deep eutectic solvent) modified magnetic silica

  19. Application of an electrochemical immunosensor with a MWCNT/PDAA modified electrode for detection of serum trypsin.

    Science.gov (United States)

    Yi, Qiang; Liu, Qicai; Gao, Feng; Chen, Qingquan; Wang, Guina

    2014-06-10

    To establish an electrochemical immunosensor for the determination of serum trypsin levels using a multiwall carbon nanotubes (MWCNTs)-composite-modified electrode. A MWCNT composite coated on the surface of bare gold electrodes was used for fixation of an anti-trypsin antibody. The assembly process and the performance indicators, including sensitivity, linear range of detection, anti-jamming performance, and stability, of the electrochemical immunosensor were examined by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). With optimized experimental conditions, the difference of the current value measured by differential pulse voltammetry (DPV) showed a linear relationship with the concentration of serum trypsin within 0.10-100 ng/mL. The detection limit for trypsin using this sensor was 0.002 ng/mL. The electrochemical immunosensor built using the MWCNT-composite-modified electrode is simple to operate and has a fast response time, along with a wide linear range, high sensitivity, and accuracy, making it suitable for serum trypsin detection.

  20. Protease-activated receptor-2 expression and the role of trypsin in cell proliferation in human pancreatic cancers.

    Science.gov (United States)

    Ohta, Tetsuo; Shimizu, Koichi; Yi, Shuangqin; Takamura, Hiroyuki; Amaya, Kohji; Kitagawa, Hirohisa; Kayahara, Masato; Ninomiya, Itasu; Fushida, Sachio; Fujimura, Takashi; Nishimura, Gen-Ichi; Miwa, Koichi

    2003-07-01

    Protease-activated receptor (PAR)-2 is a G protein-coupled receptor that is activated by trypsin. The purpose of this study was to examine PAR-2 expression and the role of trypsin in cell proliferation in human pancreatic cancer cells. All four pancreatic cancer cell lines studied, from well to undifferentiated types, AsPC-1, BxPC-3, Panc-1, and MIAPaCa-2, had significant levels of PAR-2 mRNA detected by reverse transcription-polymerase chain reaction, and showed a band of about 55 kDa corresponding to the known molecular weight of PAR-2: AsPC-1, BxPC-3 and Panc-1 showed a strong band, and MIAPaCa-2 showed a weak one. Immunocytochemically, AsPC-1, BxPC-3, and Panc-1 showed intense immunostaining for PAR-2, predominantly in the plasma membrane, while in MIAPaCa-2, immunostaining was weak. Proliferative activity of AsPC-1 cells was increased by concentrations of trypsin as low as 10 nM, and activity peaked at a concentration of 100 nM, representing almost 60% of that induced by 10% fetal bovine serum. In contrast, trypsin had no significant effect on proliferation of MIAPaCa-2 cells. These findings suggest that trypsin plays a role in the growth of PAR-2-positive pancreatic cancer cells and serves as a potent mitogen in vitro, functioning as a growth factor.

  1. A trypsin inhibitor from rambutan seeds with antitumor, anti-HIV-1 reverse transcriptase, and nitric oxide-inducing properties.

    Science.gov (United States)

    Fang, Evandro Fei; Ng, Tzi Bun

    2015-04-01

    Nephelium lappaceum L., commonly known as "rambutan," is a typical tropical tree and is well known for its juicy and sweet fruit which has an exotic flavor. Chemical studies on rambutan have led to the identification of various components such as monoterpene lactones and volatile compounds. Here, a 22.5-kDa trypsin inhibitor (N . lappaceum trypsin inhibitor (NLTI)) was isolated from fresh rambutan seeds using liquid chromatographical techniques. NLTI reduced the proteolytic activities of both trypsin and α-chymotrypsin. Dithiothreitol reduced the trypsin inhibitory activity of NLTI at a concentration of 1 mM, indicating that an intact disulfide bond is essential to the activity. NLTI inhibited HIV-1 reverse transcriptase with an IC50 of 0.73 μM. In addition, NLTI manifested a time- and dose-dependent inhibitory effect on growth in many tumor cells. NLTI is one of the few trypsin inhibitors with nitric oxide-inducing activity and may find application in tumor therapy.

  2. The properties of covalently immobilized trypsin on soap-free P(MMA-EA-AA) latex particles.

    Science.gov (United States)

    Kang, Kai; Kan, Chengyou; Yeung, Anthony; Liu, Deshan

    2005-04-19

    The covalent immobilization of trypsin onto poly[(methyl methacrylate)-co-(ethyl acrylate)-co-(acrylic acid)] latex particles, produced by a soap-free emulsion polymerization technique, was carried out using the carbodiimide method. The catalytic properties and kinetic parameters, as well as the stability of the immobilized enzyme were compared to those of the free enzyme. Results showed that the optimum temperature and pH for the immobilized trypsin in the hydrolysis of casein were 55 degrees C and 8.5, both of which were higher than that of the free form. It was found that K(m) (Michaelis constant) was 45.7 mg . ml(-1) and V(max) (maximal reaction rate) was 793.0 microg . min(-1) for immobilized trypsin, compared to a K(m) of 30.0 mg . ml(-1) and a V(max) of 5 467.5 microg . min(-1) for free trypsin. The immobilized trypsin exhibited much better thermal and chemical stabilities than its free counterpart and maintained over 63% of its initial activity after reusing ten times.

  3. Application of an Electrochemical Immunosensor with a MWCNT/PDAA Modified Electrode for Detection of Serum Trypsin

    Directory of Open Access Journals (Sweden)

    Qiang Yi

    2014-06-01

    Full Text Available Objective: To establish an electrochemical immunosensor for the determination of serum trypsin levels using a multiwall carbon nanotubes (MWCNTs-composite-modified electrode. Method: A MWCNT composite coated on the surface of bare gold electrodes was used for fixation of an anti-trypsin antibody. The assembly process and the performance indicators, including sensitivity, linear range of detection, anti-jamming performance, and stability, of the electrochemical immunosensor were examined by cyclic voltammetry (CV and electrochemical impedance spectroscopy (EIS. Results: With optimized experimental conditions, the difference of the current value measured by differential pulse voltammetry (DPV showed a linear relationship with the concentration of serum trypsin within 0.10–100 ng/mL. The detection limit for trypsin using this sensor was 0.002 ng/mL. Conclusions: The electrochemical immunosensor built using the MWCNT-composite-modified electrode is simple to operate and has a fast response time, along with a wide linear range, high sensitivity, and accuracy, making it suitable for serum trypsin detection.

  4. Shape-controlled synthesis of protein-conjugated CdS nanocrystals (NCs) and study on the binding of Cd2+/CdS to trypsin

    Science.gov (United States)

    Qin, Dezhi; Wang, Li; Wang, Yabo; Du, Xian; Zhang, Li; Zhang, Qiuxia; He, Bingyu

    2017-07-01

    Protein-conjugated CdS nanocrystals (NCs) with different morphology have been synthesized under biomimetic condition using trypsin as capping agent in aqueous medium. The reaction parameters including concentration of trypsin, pH value, reaction time, and temperature have a major influence on the morphology and optical property of CdS NCs. XRD, selected area electron diffraction (SAED), TEM, HRTEM, and EDS characterizations were used to investigate the structure, composition, morphology, and size of as-prepared products. The binding reaction between Cd2+/CdS and trypsin was investigated systematically through various spectroscopic methods. UV-vis, FT-IR, photoluminescence (PL) spectra, and conductivity analysis of Cd2+-trypsin suggest that Cd2+ ions could coordinate with the functional groups of trypsin and induce the formation of unfolding and loosening structure in protein molecules, and the change of protein conformation was also verified by circular dichroism (CD) spectra. This interaction increased local supersaturation of Cd2+ ions around the groups of trypsin and reduced the nucleation activation energy of CdS nuclei, which favored heterogeneous nucleation in trypsin matrix and resulted in the formation of inorganic-organic hybrid materials. The functional integrity of the enzyme conjugated to CdS NCs was studied by monitoring the enzymatic activity of CdS-trypsin conjugates. The fluorescence of CdS NCs is dependent strongly on trypsin which passivates the surface of NCs.

  5. A Randomized, Clinical Trial to Evaluate Efficacy and Tolerability of Trypsin:Chymotrypsin as Compared to Serratiopeptidase and Trypsin:Bromelain:Rutoside in Wound Management.

    Science.gov (United States)

    Chandanwale, Ajay; Langade, Deepak; Sonawane, Dheeraj; Gavai, Piyush

    2017-01-01

    Systemic enzyme therapy can play an important role in maintaining normal inflammatory processes within the body and thereby helps support and speed up healing. In the course of the anti-inflammatory action, enzymes degrade damaged cells and necrotic material and, through the inactivation of mediators and toxic products, they restrict the edema and pain. The study conducted at Grant Medical College, Mumbai, India was a clinical trial comparing the efficacy and tolerability of three oral enzyme treatment groups-oral tablets containing trypsin:chymotrypsin (TC) (Chymoral Forte®), serratiopeptidase (S) 5 mg oral tablets, and oral enzyme tablets containing trypsin 48 mg, bromelain 90 mg, and rutoside 100 mg (TBR)-to evaluate their healing potential in surgical wounds after orthopedic surgery. A total of 75 patients were screened, randomized, and divided into three groups in 1:1:1 ratio receiving either of the three treatments. In the TC group, erythema was significantly reduced from 3.44 on day 3 to 1.16 on day 10 (p < 0.01). There was significantly better reduction in erythema scores in the TC group as compared to S and TBR groups (p < 0.05) at each follow-up visit. Similarly reduction in the local irritation, wound discharge, edema, induration, and tenderness score with TC treatment at the end of the study was significantly higher than that observed in the other two groups. In addition TC showed significant reduction in pain on the VAS scale (p < 0.01). Global assessment of response to therapy for efficacy and tolerability was reported to be good to excellent in 88% and 92% of the patients on TC as compared to 12% and 8% with S and 12% and 8% with TBR. TC provides a better resolution of symptoms of inflammation after orthopedic surgery as compared to S and TBR, thus facilitating better wound healing. Further studies are warranted to confirm the findings. Clinical Trial Registry of India (Reg. No. CTRI/2011/07/001920).

  6. Immune Cells in Blood Recognize Tumors

    Science.gov (United States)

    NCI scientists have developed a novel strategy for identifying immune cells circulating in the blood that recognize specific proteins on tumor cells, a finding they believe may have potential implications for immune-based therapies.

  7. Social cognition: feeling voices to recognize emotions.

    Science.gov (United States)

    Adolphs, Ralph

    2010-12-21

    Our understanding of how we simulate other people's actions and feelings to recognize their emotional states is extended by a new study which finds that premotor and somatosensory cortices are required to process the emotional meaning of sounds.

  8. How plants recognize pathogens and defend themselves

    NARCIS (Netherlands)

    Wit, de P.J.G.M.

    2007-01-01

    Plants have an innate immunity system to defend themselves against pathogens. With the primary immune system, plants recognize microbe-associated molecular patterns (MAMPs) of potential pathogens through pattern recognition receptors (PRRs) that mediate a basal defense response. Plant pathogens

  9. Vegetative storage protein with trypsin inhibitor activity occurs in Sapindus mukorassi, a sapindaceae deciduous tree.

    Science.gov (United States)

    Liu, Shi-Biao; Wang, Xu-Chu; Shi, Min-Jing; Chen, Yue-Yi; Hu, Zheng-Hai; Tian, Wei-Min

    2009-04-01

    A vegetative storage protein (VSP) with trypsin inhibitor activity in a deciduous tree, Sapindus mukorassi, was characterized by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western-blot, immuno-histochemical localization, light- and electro-microscopy, together with analysis of proteinase inhibitor activity of the purified VSP in vitro. There were two proteins with molecular masses of about 23 and 27 kDa in a relatively high content in the bark tissues of terminal branches of S. mukorassi in leafless periods. The proteins decreased markedly during young shoot development, indicating their role in seasonal nitrogen storage. Immuno-histochemical localization with the polyclonal antibodies raised against the 23 kDa protein demonstrated that the 23 kDa protein was the major component of protein inclusions in protein-storing cells. The protein inclusions were identified by protein-specific staining and should correspond to the electron-dense materials in different forms in the vacuoles of phloem parenchyma cells and phloem ray parenchyma cells under an electron microscope. So, the 23 kDa protein was a typical VSP in S. mukorassi. The 23 and 27 kDa proteins shared no immuno-relatedness, whereas the 23 kDa protein was immuno-related with the 22 kDa VSP in lychee and possessed trypsin inhibitor activity. The 23 kDa protein may confer dual functions: nitrogen storage and defense.

  10. CLONING AND EXPRESSING TRYPSIN MODULATING OOSTATIC FACTOR IN Chlorella desiccata TO CONTROL MOSQUITO LARVAE.

    Science.gov (United States)

    Borovsky, Dov; Sterner, Andeas; Powell, Charles A

    2016-01-01

    The insect peptide hormone trypsin modulating oostatic factor (TMOF), a decapeptide that is synthesized by the mosquito ovary and controls the translation of the gut's trypsin mRNA was cloned and expressed in the marine alga Chlorella desiccata. To express Aedes aegypti TMOF gene (tmfA) in C. desiccata cells, two plasmids (pYES2/TMOF and pYDB4-tmfA) were engineered with pKYLX71 DNA (5 Kb) carrying the cauliflower mosaic virus (CaMV) promoter 35S(2) and the kanamycin resistant gene (neo), as well as, a 8 Kb nitrate reductase gene (nit) from Chlorella vulgaris. Transforming C. desiccata with pYES2/TMOF and pYDB4-tmfA show that the engineered algal cells express TMOF (20 ± 4 μg ± SEM and 17 ± 3 μg ± SEM, respectively in 3 × 10(8) cells) and feeding the cells to mosquito larvae kill 75 and 60% of Ae. aegypti larvae in 4 days, respectively. Southern and Northern blots analyses show that tmfA integrated into the genome of C. desiccata by homologous recombination using the yeast 2 μ circle of replication and the nit in pYES2/TMOF and pYDB4-tmfA, respectively, and the transformed algal cells express tmfA transcript. Using these algal cells it will be possible in the future to control mosquito larvae in the marsh. © 2015 Wiley Periodicals, Inc.

  11. Identification of flavonoids with trypsin inhibitory activity extracted from orange peel and green tea leaves.

    Science.gov (United States)

    Shahwar, Durre; Raza, Muhammad A; Atta-Ur-Rahman

    2013-04-01

    Orange peel (Citrus sinensis) and green tea (Camellia sinensis) leaves, rich sources of food flavonoids, were analyzed for their trypsin inhibitory potential. Hesperetin, rutin and hesperidin from orange peel, and catechin from green tea leaves, were isolated and their chemical structures were analyzed. All four compounds were evaluated for their trypsin inhibitory potential. Among all the isolated compounds, rutin exhibited the highest protease inhibition activity (75.4 ± 0.9%) with IC50 = 16 ± 2 µmol L(-1), followed by catechin (65.3 ± 1.4%; IC50 = 83 ± 9 µmol L(-1)), hesperetin (62.1 ± 1.3%; IC50 = 104 ± 12 µmol L(-1)) and hesperidin (59.7 ± 1.1%; IC50 = 127 ± 14 µmol L(-1)). Lineweaver-Burk and Dixon plots and their secondary replots indicated that all four compounds possessed non-competitive inhibition. The Ki values of hesperetin, rutin, hesperidin and catechin were calculated as 90.2 ± 1.1, 17.5 ± 0.6, 84.2 ± 1.5 and 65.1 ± 1.5 µmol L(-1) respectively. The present results suggest that the four isolated flavonoids can be used as a supplement in food for the treatment of pathologies associated with the degradation of a specific protein. © 2012 Society of Chemical Industry.

  12. Kinetic Dissection of the Pre-existing Conformational Equilibrium in the Trypsin Fold*

    Science.gov (United States)

    Vogt, Austin D.; Chakraborty, Pradipta; Di Cera, Enrico

    2015-01-01

    Structural biology has recently documented the conformational plasticity of the trypsin fold for both the protease and zymogen in terms of a pre-existing equilibrium between closed (E*) and open (E) forms of the active site region. How such plasticity is manifested in solution and affects ligand recognition by the protease and zymogen is poorly understood in quantitative terms. Here we dissect the E*-E equilibrium with stopped-flow kinetics in the presence of excess ligand or macromolecule. Using the clotting protease thrombin and its zymogen precursor prethrombin-2 as relevant models we resolve the relative distribution of the E* and E forms and the underlying kinetic rates for their interconversion. In the case of thrombin, the E* and E forms are distributed in a 1:4 ratio and interconvert on a time scale of 45 ms. In the case of prethrombin-2, the equilibrium is shifted strongly (10:1 ratio) in favor of the closed E* form and unfolds over a faster time scale of 4.5 ms. The distribution of E* and E forms observed for thrombin and prethrombin-2 indicates that zymogen activation is linked to a significant shift in the pre-existing equilibrium between closed and open conformations that facilitates ligand binding to the active site. These findings broaden our mechanistic understanding of how conformational transitions control ligand recognition by thrombin and its zymogen precursor prethrombin-2 and have direct relevance to other members of the trypsin fold. PMID:26216877

  13. The control of hyperglycemia by a novel trypsin resistant oral insulin preparation in alloxan induced type I diabetic mice

    OpenAIRE

    Sarbashri Bank; Arjun Ghosh; Suman Bhattacharya; Smarajit Maiti; Khan, Gausal A.; Sinha, Asru K.

    2016-01-01

    A trypsin resistant oral insulin preparation was made by incubating insulin for 2?h at 23??C with previously boiled cow milk at 100??C that was coagulated with 0.6?M acetic acid. The precipitate was resuspended in the same volume of milk. The immunoblot analysis of the suspended proteins treated with 200?ng of trypsin/ml for 3?h demonstrated that the 80.1% of the insulin in the suspension survived the proteolytic degradation compared to 0% of the hormone survived in the control. The feeding o...

  14. Isolation and Partial Characterization of Human Amniotic Epithelial Cells: The Effect of Trypsin

    Science.gov (United States)

    Tabatabaei, Meraj; Mosaffa, Nariman; Nikoo, Shohreh; Bozorgmehr, Mahmood; Ghods, Roya; Kazemnejad, Somaieh; Rezania, Simin; Keshavarzi, Bahareh; Arefi, Soheila; Ramezani-Tehrani, Fahimeh; Mirzadegan, Ebrahim; Zarnani, Amir-Hassan

    2014-01-01

    Background Despite the extensive information available in the literature, cell surface marker signature of human Amniotic Epithelial Cells (hAECs) remains controversial. The aim of the present study was to characterize immunophenotypic features, proliferative capacity and immunogenicity of hAECs. We also tested whether expression of some cell surface markers is influenced by the type of trypsin used for tissue digestion. Methods Single cell suspensions of amniotic membranes from four human placentas were isolated by enzymatic digestion and expression of CD9, CD10, CD29, CD34, CD38, CD44, CD45, CD73, CD105, CD133, HLA-I, HLA-DR, HLA-G, SSEA-4, STRO-1 and OCT-4 was then evaluated by flow cytometry. The differential impact of four trypsin types on the yield and expression of CD105 and HLA-I was also determined. The proliferative capacity of cultured hAECs was assessed and compared in the presence and absence of Epidermal Growth Factor (EGF). To test their immunogenicity, hAECs were injected into Balb/c mice and the reactivity of hyperimmunized sera was examined by immunofluorescence staining. Results Nearly all purified cells expressed mesenchymal markers, CD9, CD10, CD29, and CD73 and the embryonic marker, SSEA-4. A large proportion of the cells also expressed STRO-1 and OCT-4. The purified cells also expressed HLA-G and HLA-I. A very small proportion of hAECs expressed CD34, CD38, CD44, CD133 and HLA-DR. The type of trypsin used for enzymatic digestion affected both the percentage and expression of HLA-I and CD105. hAECs revealed substantial proliferative capacity only when cultured in the medium supplemented with EGF. These cells were shown to be capable of inducing high amounts of anti-donor antibodies. Conclusion Here we provided evidence that hAECs are immunogenic cells with high level of HLA-I expression. Furthermore, this work highlighted the impact of isolation procedure on the immunophenotype of hAEC. PMID:24523953

  15. Probing the binding mechanisms of α-tocopherol to trypsin and pepsin using isothermal titration calorimetry, spectroscopic, and molecular modeling methods.

    Science.gov (United States)

    Li, Xiangrong; Ni, Tianjun

    2016-06-01

    α-Tocopherol is a required nutrient for a variety of biological functions. In this study, the binding of α-tocopherol to trypsin and pepsin was investigated using isothermal titration calorimetry (ITC), steady-state and time-resolved fluorescence measurements, circular dichroism (CD) spectroscopy, and molecular modeling methods. Thermodynamic investigations reveal that α-tocopherol binds to trypsin/pepsin is synergistically driven by enthalpy and entropy. The fluorescence experimental results indicate that α-tocopherol can quench the fluorescence of trypsin/pepsin through a static quenching mechanism. The binding ability of α-tocopherol with trypsin/pepsin is in the intermediate range, and one molecule of α-tocopherol combines with one molecule of trypsin/pepsin. As shown by circular dichroism (CD) spectroscopy, α-tocopherol may induce conformational changes of trypsin/pepsin. Molecular modeling displays the specific binding site and gives information about binding forces and α-tocopherol-tryptophan (Trp)/tyrosine (Tyr) distances. In addition, the inhibition rate of α-tocopherol on trypsin and pepsin was studied. The study provides a basic data set for clarifying the binding mechanisms of α-tocopherol with trypsin and pepsin and is helpful for understanding its biological activity in vivo.

  16. Postpartum Mood Disorders: Recognizing the symptoms

    OpenAIRE

    Misri, Shaila; Burgmann, Allan J.F.

    1992-01-01

    Adjusting to the role of mother, a creative and joyous change for most women, combines with simultaneous physiological and psychological changes to develop into psychiatric problems in some women. Three common syndromes during the postpartum period are postpartum blues, postpartum depression, and postpartum psychosis. Any postpartum condition should be diagnosed rapidly to prevent short- and long-term disorders.

  17. Recognizing, Confronting, and Eliminating Workplace Bullying.

    Science.gov (United States)

    Berry, Peggy Ann; Gillespie, Gordon L; Fisher, Bonnie S; Gormley, Denise K

    2016-07-01

    Workplace bullying (WPB) behaviors negatively affect nurse productivity, satisfaction, and retention, and hinder safe patient care. The purpose of this article is to define WPB, differentiate between incivility and WPB, and recommend actions to prevent WPB behaviors. Informed occupational and environmental health nurses and nurse leaders must recognize, confront, and eliminate WPB in their facilities and organizations. Recognizing, confronting, and eliminating WPB behaviors in health care is a crucial first step toward sustained improvements in patient care quality and the health and safety of health care employees. © 2016 The Author(s).

  18. Recognizing textual entailment models and applications

    CERN Document Server

    Dagan, Ido; Sammons, Mark

    2013-01-01

    In the last few years, a number of NLP researchers have developed and participated in the task of Recognizing Textual Entailment (RTE). This task encapsulates Natural Language Understanding capabilities within a very simple interface: recognizing when the meaning of a text snippet is contained in the meaning of a second piece of text. This simple abstraction of an exceedingly complex problem has broad appeal partly because it can be conceived also as a component in other NLP applications, from Machine Translation to Semantic Search to Information Extraction. It also avoids commitment to any sp

  19. Cyclic RGD peptide conjugated trypsin etched gold quantum clusters: novel biolabeling agents for stem cell imaging.

    Science.gov (United States)

    Das, Bodhisatwa; Fernandez, Francis; John, Annie; Sharma, Chandra P

    2012-01-01

    Quantum clusters are sub-nano sized materials mostly synthesized from noble metals with luminescence property and high quantum yield. They are important to biomedical scientists because of their excellent optical properties. Here we represent a tool for cell imaging purpose using protein stabilized gold quantum clusters. Intestinal protease Trypsin was used to develop clusters. They were conjugated to cyclic RGD peptides by EDAC coupling. Cell imaging property was checked by transfecting the RGD-conjugated quantum clusters to bone marrow stem cells. For characterization of RGD-conjugated quantum clusters UV-Vis, Fluorescence and FTIR spectroscopy was performed. DLS and Zeta potential measurement also have been done. To check the bio compatibility of the quantum clusters MTT assay, AFM and blood cell adhesion study were performed. The samples are found out to be good for cell imaging as well as bio compatible and hemo-compatible.

  20. High-gradient magnetic affinity separation of trypsin from porcine pancreatin

    DEFF Research Database (Denmark)

    Hubbuch, Jürgen; Thomas, Owen R. T.

    2002-01-01

    We introduce a robust and scale-flexible approach to macromolecule purification employing tailor-made magnetic adsorbents and high-gradient magnetic separation technology adapted from the mineral processing industries. Detailed procedures for the synthesis of large quantities of low-cost defined...... increased scale using a high-gradient magnetic separation system to capture loaded benzamidine-linked adsorbents following batch adsorption. With the aid of a simple recycle loop over 80% of the initially adsorbed trypsin was recovered in-line with an overall purification factor of approximate to3.5....... submicron-sized magnetic supports are presented. These support materials exhibit unique features, which facilitate their large-scale processing using high magnetic field gradients, namely sufficiently high magnetization, a relatively narrow particle size distribution and ideal superparamagnetism. Following...

  1. Trypsin, Tryptase, and Thrombin Polarize Macrophages towards a Pro-Fibrotic M2a Phenotype.

    Directory of Open Access Journals (Sweden)

    Michael J V White

    Full Text Available For both wound healing and the formation of a fibrotic lesion, circulating monocytes enter the tissue and differentiate into fibroblast-like cells called fibrocytes and pro-fibrotic M2a macrophages, which together with fibroblasts form scar tissue. Monocytes can also differentiate into classically activated M1 macrophages and alternatively activated M2 macrophages. The proteases thrombin, which is activated during blood clotting, and tryptase, which is released by activated mast cells, potentiate fibroblast proliferation and fibrocyte differentiation, but their effect on macrophages is unknown. Here we report that thrombin, tryptase, and the protease trypsin bias human macrophage differentiation towards a pro-fibrotic M2a phenotype expressing high levels of galectin-3 from unpolarized monocytes, or from M1 and M2 macrophages, and that these effects appear to operate through protease-activated receptors. These results suggest that proteases can initiate scar tissue formation by affecting fibroblasts, fibrocytes, and macrophages.

  2. The starch-bound alpha-amylase/trypsin-inhibitors in Avena.

    Science.gov (United States)

    Gazza, Laura; Gazzelloni, Gloria; Taddei, Federica; Latini, Arianna; Muccilli, Vera; Alfieri, Michela; Conti, Salvatore; Redaelli, Rita; Pogna, Norberto E

    2016-12-01

    Oat kernels exhibit an extra-soft texture, a trait recently demonstrated to be largely modulated by starch-bound tryptophan-rich 2S proteins, the vromindolines. In this study, fractionation by two-dimensional electrophoresis of starch-bound proteins in 25 oat (Avena sativa) cultivars and 11 diploid or tetraploid Avena species revealed novel 2S proteins called Avena α-amylase/trypsin-inhibitors (AATI) because of their sequence similarity with wheat α-amylase/trypsin inhibitors. Thirty-seven AATI polypeptides, about 14 kDa in size, were split into three families named AATI-1, AATI-2, and AATI-3 with different primary structures and isoelectric points. AATI-1 and AATI-2 proteins showed 55.5-60.0 % sequence similarity with wheat α-amylase inhibitors CM1, CM2, and CM16, which have been found to cause innate immunity responses in celiac disease and non-celiac gluten sensitivity. Diploid A-genome and tetraploid AC-genome oat species possess three and five genes encoding for the AATI proteins, respectively, whereas hexaploid A. sativa exhibits 12 genes dispersed over the A-, C-, and D-genomes. Some AATI proteins expressed in hexaploid oats were assigned to the A-genome based on similarity to their counterparts in diploid species, contributing to further clarify the genetic origin of hexaploid oats. Moreover, AATI may interact with starch-bound vromindolines in determining the extra-soft texture of oat kernels and, due to their balanced amino acid compositions, may contribute to the biological value of oat proteins in a positive manner.

  3. Characterization of Clostridium perfringens TpeL Toxin Gene Carriage, Production, Cytotoxic Contributions, and Trypsin Sensitivity

    Science.gov (United States)

    Chen, Jianming

    2015-01-01

    Large clostridial toxins (LCTs) are produced by at least four pathogenic clostridial species, and several LCTs are proven pivotal virulence factors for both human and veterinary diseases. TpeL is a recently identified LCT produced by Clostridium perfringens that has received relatively limited study. In response, the current study surveyed carriage of the tpeL gene among different C. perfringens strains, detecting this toxin gene in some type A, B, and C strains but not in any type D or E strains. This study also determined that all tested strains maximally produce, and extracellularly release, TpeL at the late-log or early-stationary growth stage during in vitro culture, which is different from the maximal late-stationary-phase production reported previously for other LCTs and for TpeL production by C. perfringens strain JIR12688. In addition, the present study found that TpeL levels in culture supernatants can be repressed by either glucose or sucrose. It was also shown that, at natural production levels, TpeL is a significant contributor to the cytotoxic activity of supernatants from cultures of tpeL-positive strain CN3685. Lastly, this study identified TpeL, which presumably is produced in the intestines during diseases caused by TpeL-positive type B and C strains, as a toxin whose cytotoxicity decreases after treatment with trypsin; this finding may have pathophysiologic relevance by suggesting that, like beta toxin, TpeL contributes to type B and C infections in hosts with decreased trypsin levels due to disease, diet, or age. PMID:25824828

  4. Prediction of trypsin/molecular fragment binding affinities by free energy decomposition and empirical scores

    Science.gov (United States)

    Benson, Mark L.; Faver, John C.; Ucisik, Melek N.; Dashti, Danial S.; Zheng, Zheng; Merz, Kenneth M.

    2012-05-01

    Two families of binding affinity estimation methodologies are described which were utilized in the SAMPL3 trypsin/fragment binding affinity challenge. The first is a free energy decomposition scheme based on a thermodynamic cycle, which included separate contributions from enthalpy and entropy of binding as well as a solvent contribution. Enthalpic contributions were estimated with PM6-DH2 semiempirical quantum mechanical interaction energies, which were modified with a statistical error correction procedure. Entropic contributions were estimated with the rigid-rotor harmonic approximation, and solvent contributions to the free energy were estimated with several different methods. The second general methodology is the empirical score LISA, which contains several physics-based terms trained with the large PDBBind database of protein/ligand complexes. Here we also introduce LISA+, an updated version of LISA which, prior to scoring, classifies systems into one of four classes based on a ligand's hydrophobicity and molecular weight. Each version of the two methodologies (a total of 11 methods) was trained against a compiled set of known trypsin binders available in the Protein Data Bank to yield scaling parameters for linear regression models. Both raw and scaled scores were submitted to SAMPL3. Variants of LISA showed relatively low absolute errors but also low correlation with experiment, while the free energy decomposition methods had modest success when scaling factors were included. Nonetheless, re-scaled LISA yielded the best predictions in the challenge in terms of RMS error, and six of these models placed in the top ten best predictions by RMS error. This work highlights some of the difficulties of predicting binding affinities of small molecular fragments to protein receptors as well as the benefit of using training data.

  5. Trypsin Isoinhibitors with Antiproliferative Activity toward Leukemia Cells from Phaseolus vulgaris cv “White Cloud Bean”

    Directory of Open Access Journals (Sweden)

    Jian Sun

    2010-01-01

    Full Text Available A purification protocol that comprised ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on SP-Sepharose, and gel filtration by FPLC on Superdex 75 was complied to isolate two trypsin inhibitors from Phaseolus vulgaris cv “White Cloud Bean”. Both trypsin inhibitors exhibited a molecular mass of 16 kDa and reduced the activity of trypsin with an IC50 value of about 0.6 M. Dithiothreitol attenuated the trypsin inhibitory activity, signifying that an intact disulfide bond is indispensable to the activity. [Methyl-3H] thymidine incorporation by leukemia L1210 cells was inhibited with an IC50 value of 28.8 M and 21.5 M, respectively. They were lacking in activity toward lymphoma MBL2 cells and inhibitory effect on HIV-1 reverse transcriptase and fungal growth when tested up to 100 M.

  6. A comparative study of trypsin specificity based on QM/MM molecular dynamics simulation and QM/MM GBSA calculation.

    Science.gov (United States)

    Chen, Jianzhong; Wang, Jinan; Zhang, Qinggang; Chen, Kaixian; Zhu, Weiliang

    2015-01-01

    Hydrogen bonding and polar interactions play a key role in identification of protein-inhibitor binding specificity. Quantum mechanics/molecular mechanics molecular dynamics (QM/MM MD) simulations combined with DFT and semi-empirical Hamiltonian (AM1d, RM1, PM3, and PM6) methods were performed to study the hydrogen bonding and polar interactions of two inhibitors BEN and BEN1 with trypsin. The results show that the accuracy of treating the hydrogen bonding and polar interactions using QM/MM MD simulation of PM6 can reach the one obtained by the DFT QM/MM MD simulation. Quantum mechanics/molecular mechanics generalized Born surface area (QM/MM-GBSA) method was applied to calculate binding affinities of inhibitors to trypsin and the results suggest that the accuracy of binding affinity prediction can be significantly affected by the accurate treatment of the hydrogen bonding and polar interactions. In addition, the calculated results also reveal the binding specificity of trypsin: (1) the amidinium groups of two inhibitors generate favorable salt bridge interaction with Asp189 and form hydrogen bonding interactions with Ser190 and Gly214, (2) the phenyl of inhibitors can produce favorable van der Waals interactions with the residues His58, Cys191, Gln192, Trp211, Gly212, and Cys215. This systematic and comparative study can provide guidance for the choice of QM/MM MD methods and the designs of new potent inhibitors targeting trypsin.

  7. Protease-activated receptor-2 regulates trypsin expression in the brain and protects against seizures and epileptogenesis.

    Science.gov (United States)

    Lohman, Rink-Jan; O'Brien, Terence J; Cocks, Thomas M

    2008-04-01

    Protease-activated receptor-2 (PAR(2)), primarily involved in inflammation, is highly expressed in limbic regions of the brain such as the hippocampus. Although extracellular proteolysis is involved in normal and stress-related neuronal plasticity associated with learning, memory and inflammatory disease states, little is known about the role of PAR(2) and its physiological agonist, trypsin, in the brain. We show immunohistochemically that trypsin co-localises with tissue plasminogen activator within granular-like structures in PAR(2)-positive pyramidal neurons of the rat hippocampus. Central administration of the PAR(2) peptide agonist, SLIGRL, inhibited electrical amygdala kindling-induced epileptogenesis and abolished kindling-induced over-expression of trypsin in the hippocampus. SLIGRL similarly attenuated kindling when administered subcutaneously. Non-enzymatic activation of neuronal PAR(2) using SLIGRL may thus activate feedback mechanisms to inhibit the over-production of trypsin and possibly other proteases during brain insults and thereby attenuate pathogenesis. Prophylactic systemic administration of non-proteolytic PAR(2) agonists may therefore represent a novel approach to protect against epileptogenic brain insults.

  8. Kunitz trypsin inhibitor in addition to Bowman-Birk inhibitor influence stability of lunasin against pepsin-pancreatin hydrolysis

    Science.gov (United States)

    Soybean contains several biologically active components and one of this belongs to the bioactive peptide group. The objectives of this study were to produce different lunasin-enriched preparations (LEP) and determine the effect of Bowman-Birk inhibitor and Kunitz trypsin concentrations on the stabil...

  9. Cis-mediated down-regulation of a trypsin gene associated with Bt resistance in cotton bollworm.

    Science.gov (United States)

    Liu, Chenxi; Xiao, Yutao; Li, Xianchun; Oppert, Brenda; Tabashnik, Bruce E; Wu, Kongming

    2014-11-27

    Transgenic plants producing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) are useful for pest control, but their efficacy is reduced when pests evolve resistance. Here we examined the mechanism of resistance to Bt toxin Cry1Ac in the laboratory-selected LF5 strain of the cotton bollworm, Helicoverpa armigera. This strain had 110-fold resistance to Cry1Ac protoxin and 39-fold resistance to Cry1Ac activated toxin. Evaluation of five trypsin genes revealed 99% reduced transcription of one trypsin gene (HaTryR) was associated with resistance. Silencing of this gene with RNA interference in susceptible larvae increased their survival on diets containing Cry1Ac. Bioassays of progeny from crosses revealed that resistance to Cry1Ac was genetically linked with HaTryR. We identified mutations in the promoter region of HaTryR in the resistant strain. In transfected insect cell lines, transcription was lower when driven by the resistant promoter compared with the susceptible promoter, implicating cis-mediated down-regulation of HaTryR transcription as a mechanism of resistance. The results suggest that H. armigera can adapt to Bt toxin Cry1Ac by decreased expression of trypsin. Because trypsin activation of protoxin is a critical step in toxicity, transgenic plants with activated toxins rather than protoxins might increase the durability of Bt crops.

  10. Cloning and characterization of trypsin- and chymotrypsin-like genes in the striped rice stem borer, Chilo suppressalis.

    Science.gov (United States)

    Ge, Zhao-Yu; Wan, Pin-Jun; Han, Zhao-Jun

    2012-04-01

    Serine proteinases including trypsins and chymotrypsins play various important roles in insects, including food digestion, immune defense, and zymogen activation. Studies on insect serine proteinases could reveal their feeding preference (polyphagous and monophagous) and facilitate identification of protease inhibitors, which can be engineered for pest management. In this paper, 11 transcripts encoding trypsin- and chymotrypsin-like proteins were cloned from the striped rice stem borer, Chilo suppressalis (Walker). All the predicted proteins share high sequence similarity with known trypsin- and chymotrypsin-like proteins from either lepidopterans or dipterans, and most of the proteins have conserved motifs that are characteristics of serine proteinases. Among the 11 cloned genes, six were expressed predominantly and one exclusively in the midgut of the insect, three were expressed relatively evenly in examined tissues, and one was not expressed in either the gut or hemolymph based on RT-PCR results. The seven genes that were predominantly or exclusively expressed in the gut were also affected by feeding on different host plants. The genes that were expressed in the gut and were affected by host plants are likely to encode digestive proteinases. The identification of trypsin- and chymotrypsin-like genes in this insect species is the first step towards further comparative studies and for identification of insect-specific proteinase inhibitors, which might be engineered to protect rice plants against the striped rice stem borer, which is one of the destructive pests of rice.

  11. Cold adaptation, ca2+ dependency and autolytic stability are related features in a highly active cold-adapted trypsin resistant to autoproteolysis engineered for biotechnological applications.

    Directory of Open Access Journals (Sweden)

    Alvaro Olivera-Nappa

    Full Text Available Pig trypsin is routinely used as a biotechnological tool, due to its high specificity and ability to be stored as an inactive stable zymogen. However, it is not an optimum enzyme for conditions found in wound debriding for medical uses and trypsinization processes for protein analysis and animal cell culturing, where low Ca(2+ dependency, high activity in mild conditions and easy inactivation are crucial. We isolated and thermodynamically characterized a highly active cold-adapted trypsin for medical and laboratory use that is four times more active than pig trypsin at 10(° C and at least 50% more active than pig trypsin up to 50(° C. Contrary to pig trypsin, this enzyme has a broad optimum pH between 7 and 10 and is very insensitive to Ca(2+ concentration. The enzyme is only distantly related to previously described cryophilic trypsins. We built and studied molecular structure models of this trypsin and performed molecular dynamic calculations. Key residues and structures associated with calcium dependency and cryophilicity were identified. Experiments indicated that the protein is unstable and susceptible to autoproteolysis. Correlating experimental results and structural predictions, we designed mutations to improve the resistance to autoproteolysis and conserve activity for longer periods after activation. One single mutation provided around 25 times more proteolytic stability. Due to its cryophilic nature, this trypsin is easily inactivated by mild denaturation conditions, which is ideal for controlled proteolysis processes without requiring inhibitors or dilution. We clearly show that cold adaptation, Ca(2+ dependency and autolytic stability in trypsins are related phenomena that are linked to shared structural features and evolve in a concerted fashion. Hence, both structurally and evolutionarily they cannot be interpreted and studied separately as previously done.

  12. Fast Feature-Recognizing Optoelectronic System

    Science.gov (United States)

    Thakoor, S.; Thakoor, A. P.

    1990-01-01

    Proposed optoelectronic system recognizes features or classifies images by processing outputs of photosensors rapidly, in parallel, through circuits developed in research on neural networks. Array of photoconductive elements serve as photomodulated connections in electronic neural network, which provides high speed data compression to generate feature vector. System able to "learn" new patterns for subsequent recognition. Potential applications in robotic vision systems and pattern recognition.

  13. Great Apes' Capacities to Recognize Relational Similarity

    Science.gov (United States)

    Haun, Daniel B. M.; Call, Josep

    2009-01-01

    Recognizing relational similarity relies on the ability to understand that defining object properties might not lie in the objects individually, but in the relations of the properties of various object to each other. This aptitude is highly relevant for many important human skills such as language, reasoning, categorization and understanding…

  14. How should a speech recognizer work?

    NARCIS (Netherlands)

    Scharenborg, O.E.; Norris, D.; Bosch, L.F.M. ten; McQueen, J.M.

    2005-01-01

    Although researchers studying human speech recognition (HSR) and automatic speech recognition (ASR) share a common interest in how information processing systems (human or machine) recognize spoken language, there is little communication between the two disciplines. We suggest that this lack of

  15. How State Laws Recognize Advanced Nursing Practice.

    Science.gov (United States)

    Trandel-Korenchuk, Darlene M.; Trandel-Korenchuk, Keith M.

    1978-01-01

    Reviews modifications in state health practice statutes to recognize the expanded scope of nursing practice in view of the disparity between medical functions actually performed by nurses and those considered within the legal definition. Various state approaches indicate a trend to give legal validity to acts performed by nurses. (MF)

  16. Recognizing primary emotions in communication in adults with intellectual disability

    Directory of Open Access Journals (Sweden)

    Đorđević Mirjana

    2017-01-01

    Full Text Available The ability to recognize emotions helps people understand social situations and plan their own behavior in specific social contexts. Paralinguistic segments of communication (facial expression and prosodic characteristics significantly contribute to drawing conclusions on the speaker's emotions, and according to some authors, they are considered even more dominant and more reliable than linguistic segments in decoding emotions tasks. The aim of this paper was to determine the ability to recognize primary emotions in communication with the help of paralinguistic indicators, in adults with mild and moderate intellectual disability. The sample consisted of 60 adult participants with mild (N=25 and moderate (N=35 intellectual disability, equal with regard to gender and type of housing. The participants with mild intellectual disability were aged 20-55 (M=31.24; SD=8.84, while those with moderate intellectual disability were 22-55 years of age (M=34.17; SD=8.76. The Assessment Battery for Communication, ABaCo, (Sacco et al., 2008 was used for assessing the ability to understand emotions in communication. The obtained results showed that adults with intellectual disability experienced most difficulties in detecting fear. By applying the T test for independent samples, it was determined that there were no statistically significant differences between men and women both with mild and moderate ID with regard to their ability to recognize emotions (p>0,05. With regard to the type of housing, statistically significant differences were obtained only in the group of participants with moderate intellectual disability in detecting fear (t (33=-3.51; p=0.01, in favor of those who lived in institutions. The length of living in an institution positively and moderately correlated with the ability to recognize anger in adults with intellectual disability (r=0.46, p<0.05.

  17. Purification, characterization and cDNA cloning of a trypsin from the hepatopancreas of snakehead (Channa argus).

    Science.gov (United States)

    Zhou, Long-Zhen; Ruan, Mi-Mi; Cai, Qiu-Feng; Liu, Guang-Ming; Sun, Le-Chang; Su, Wen-Jin; Cao, Min-Jie

    2012-03-01

    A trypsin was purified from the hepatopancreas of snakehead (Channa argus) by ammonium sulfate fractionation and a series of column chromatographies including DEAE-Sepharose, Sephacryl S-200 HR and Hi-Trap Capto-Q. The molecular mass of the purified trypsin was about 22 kDa, as estimated by SDS-PAGE. The optimum pH and temperature of the purified trypsin were 9.0 and 40°C, respectively. The trypsin was stable in the pH range of 7.5-9.5 and below 45°C. The enzymatic activity was strongly inhibited by serine proteinase inhibitors, such as MBTI, Pefabloc SC, PMSF, LBTI and benzamidine. Peptide mass fingerprinting (PMF) of the purified protein obtained 2 peptide fragments with 25 amino acid residues and were 100% identical to the trypsinogen from pufferfish (Takifugu rubripes). The activation energy (Ea) of this enzyme was 24.65 kJ·M(-1). Apparent K(m) was 1.02 μM and k(cat) was 148 S(-1) for fluorogenic substrate Boc-Phe-Ser-Arg-MCA. A trypsinogen gene encoding 247 amino acid residues was further cloned on the basis of the sequence obtained from PMF and the conserved site peptide of trypsinogen together with 5'-RACE and 3'-RACE. The deduced amino acid sequence contains a signal peptide of 15 residues and an activation peptide of 9 amino acid residues with a mature protein of 223 residues. The catalytic triad His-64, Asp-107, Ser-201 and 12 Cys residues which may form 6 disulfide bonds were conserved. Compared with the PMF data, only 2 amino acid residues difference were identified, suggesting the cloned trypsinogen is quite possibly the precursor of the purified trypsin. Copyright © 2011 Elsevier Inc. All rights reserved.

  18. Changes in antigenicity of porcine serum albumin in gamma-irradiated sausage extract by treatment with pepsin and trypsin

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Koth-Bong-Woo-Ri; Song, Eu-Jin [Department of Food Science and Technology/Institute of Food Science, Pukyong National University, Busan 608-737 (Korea, Republic of); Lee, So-Young [Traditional Food Research Group, Korea Food Research Institute, Seongnam 463-746 (Korea, Republic of); Park, Jin-Gyu [Radiation Food Science and Biotechnology, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongup 580-185 (Korea, Republic of); Lee, Ju-Woon [National Federation of Fisheries Cooperatives, Fisheries Economic Institute, Seoul 138-827 (Korea, Republic of); Byun, Myung-Woo [Department of Culinary Nutrition, Woosong University, Daejon 300-718 (Korea, Republic of); Ahn, Dong-Hyun, E-mail: dhahn@pknu.ac.kr [Department of Food Science and Technology/Institute of Food Science, Pukyong National University, Busan 608-737 (Korea, Republic of)

    2011-11-15

    Pork is known as an allergenic food with porcine serum albumin (PSA, 66 kDa) representing the major allergen. This study was conducted to investigate the change in antigenicity of PSA in gamma-irradiated sausage extract treated with pepsin and trypsin. Sausage products (A and B) were irradiated at 1, 3, 10, and 20 kGy. After irradiation, sausage proteins were extracted and digested with pepsin (1:200, 30 min) and trypsin (1:300, 5, 30, 60, 90, and 120 min). The binding ability of PSA in extracts of the irradiated sausages (A and B) decreased by over 3 kGy relative to the binding ability of PSA in extracts of intact sausages and showed no notable differences when the dose of radiation ranged from 3 to 20 kGy. After treatment with pepsin and trypsin, the binding ability of PSA in extracts of the irradiated sausages was decreased more relative to that of intact sausages and showed no significant differences when the period of trypsin treatment is increased or when the dose of irradiation is increased. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results indicated that there was no visible change in the intensity of the PSA band in extracts of the irradiated sausages. After pepsin and trypsin treatment, the intensity of PSA band faded with increasing doses of irradiation. In conclusion, antigenicity of PSA in pork sausages could be reduced by gamma irradiation. - Highlights: > Change in antigenicity of PSA in irradiated sausage extract (ISE) was examined. > Binding ability of PSA in ISE was decreased compared to intact extract. > Binding ability of PSA in ISE after enzyme treatments was also further decreased. > Intensity of PSA band in ISE after enzyme treatments became weak.

  19. Complete Currarino Syndrome Recognized in Adulthood

    Directory of Open Access Journals (Sweden)

    Sinan Akay

    2015-01-01

    Full Text Available Currarino syndrome is a hereditary pathology that is characterized by sacrococcygeal bone defect, presacral mass, and anorectal malformation. Sacrococcygeal bone defect is almost always a part of the syndrome. The complete form of this entity displays all three abnormalities and is very uncommon. In this report, we present the magnetic resonance imaging findings of a case with complete form of Currarino syndrome recognized in adulthood.

  20. Escherichia coli fimbriae recognizing sialyl galactosides.

    OpenAIRE

    Korhonen, T K; Väisänen-Rhen, V; Rhen, M; Pere, A; Parkkinen, J; Finne, J

    1984-01-01

    Fimbriae recognizing sialyl galactosides (S fimbriae) were purified from an Escherichia coli strain. The S fimbriae were morphologically identical to type 1 and P fimbriae of E. coli and showed a hemagglutination that was abolished when erythrocytes were treated with neuraminidase. Hemagglutination by the purified fimbriae was inhibited by orosomucoid but not by its desialylated derivative. Of the oligosaccharides tested, sialyl-(alpha 2-3)-lactose and sialyl-(alpha 2-3)-N-acetyllactosamine h...

  1. Recognizing Dynamic Faces in Malaysian Chinese Participants.

    Science.gov (United States)

    Tan, Chrystalle B Y; Sheppard, Elizabeth; Stephen, Ian D

    2016-03-01

    High performance level in face recognition studies does not seem to be replicable in real-life situations possibly because of the artificial nature of laboratory studies. Recognizing faces in natural social situations may be a more challenging task, as it involves constant examination of dynamic facial motions that may alter facial structure vital to the recognition of unfamiliar faces. Because of the incongruences of recognition performance, the current study developed stimuli that closely represent natural social situations to yield results that more accurately reflect observers' performance in real-life settings. Naturalistic stimuli of African, East Asian, and Western Caucasian actors introducing themselves were presented to investigate Malaysian Chinese participants' recognition sensitivity and looking strategies when performing a face recognition task. When perceiving dynamic facial stimuli, participants fixated most on the nose, followed by the mouth then the eyes. Focusing on the nose may have enabled participants to gain a more holistic view of actors' facial and head movements, which proved to be beneficial in recognizing identities. Participants recognized all three races of faces equally well. The current results, which differed from a previous static face recognition study, may be a more accurate reflection of observers' recognition abilities and looking strategies. © The Author(s) 2015.

  2. Quantitative Assessment of the Effects of Trypsin Digestion Methods on Affinity Purification-Mass Spectrometry-based Protein-Protein Interaction Analysis.

    Science.gov (United States)

    Zhang, Yueqing; Sun, Hong; Zhang, Jing; Brasier, Allan R; Zhao, Yingxin

    2017-08-04

    Affinity purification-mass spectrometry (AP-MS) has become the method of choice for discovering protein-protein interactions (PPIs) under native conditions. The success of AP-MS depends on the efficiency of trypsin digestion and the recovery of the tryptic peptides for MS analysis. Several different protocols have been used for trypsin digestion of protein complexes in AP-MS studies, but no systematic studies have been conducted on the impact of trypsin digestion conditions on the identification of PPIs. Here, we used NFκB/RelA and Bromodomain-containing protein 4 (BRD4) as baits and test five distinct trypsin digestion methods (two using "on-beads," three using "elution-digestion" protocols). Although the performance of the trypsin digestion protocols change slightly depending on the different baits, antibodies and cell lines used, we found that elution-digestion methods consistently outperformed on-beads digestion methods. The high-abundance interactors can be identified universally by all five methods, but the identification of low-abundance RelA interactors is significantly affected by the choice of trypsin digestion method. We also found that different digestion protocols influence the selected reaction monitoring (SRM)-MS quantification of PPIs, suggesting that optimization of trypsin digestion conditions may be required for robust targeted analysis of PPIs.

  3. Development of a rapid high-efficiency scalable process for acetylated Sus scrofa cationic trypsin production from Escherichia coli inclusion bodies.

    Science.gov (United States)

    Zhao, Mingzhi; Wu, Feilin; Xu, Ping

    2015-12-01

    Trypsin is one of the most important enzymatic tools in proteomics and biopharmaceutical studies. Here, we describe the complete recombinant expression and purification from a trypsinogen expression vector construct. The Sus scrofa cationic trypsin gene with a propeptide sequence was optimized according to Escherichia coli codon-usage bias and chemically synthesized. The gene was inserted into pET-11c plasmid to yield an expression vector. Using high-density E. coli fed-batch fermentation, trypsinogen was expressed in inclusion bodies at 1.47 g/L. The inclusion body was refolded with a high yield of 36%. The purified trypsinogen was then activated to produce trypsin. To address stability problems, the trypsin thus produced was acetylated. The final product was generated upon gel filtration. The final yield of acetylated trypsin was 182 mg/L from a 5-L fermenter. Our acetylated trypsin product demonstrated higher BAEE activity (30,100 BAEE unit/mg) than a commercial product (9500 BAEE unit/mg, Promega). It also demonstrated resistance to autolysis. This is the first report of production of acetylated recombinant trypsin that is stable and suitable for scale-up. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Atomic-scale investigation of the interactions between tetrabromobisphenol A, tetrabromobisphenol S and bovine trypsin by spectroscopies and molecular dynamics simulations

    Energy Technology Data Exchange (ETDEWEB)

    Ding, Keke [Institute of Environmental Science, College of Environmental and Resource Sciences, Zhejiang University, Hangzhou 310058 (China); Zhang, Huanxin [College of Marine Life Sciences, Ocean University of China, Qingdao 266003 (China); Wang, Haifei; Lv, Xuan; Pan, Liumeng; Zhang, Wenjing [Institute of Environmental Science, College of Environmental and Resource Sciences, Zhejiang University, Hangzhou 310058 (China); Zhejiang Provincial Key Laboratory of Organic Pollution Process and Control, Hangzhou 310058 (China); Zhuang, Shulin, E-mail: shulin@zju.edu.cn [Institute of Environmental Science, College of Environmental and Resource Sciences, Zhejiang University, Hangzhou 310058 (China); Zhejiang Provincial Key Laboratory of Organic Pollution Process and Control, Hangzhou 310058 (China)

    2015-12-15

    Highlights: • The interaction of TBBPA/TBBPS with bovine trypsin was deciphered for the first time. • The fluorescence of bovine trypsin was quenched in a concentration-dependent mode. • TBBPA and TBBPS bind at the ANS binding site with distinct binding modes. • TBBPS has a higher binding affinity toward bovine trypsin than TBBPA. • Our in vitro and in silico approach is helpful to assess risk of TBBPA-related BFRs. - Abstract: Tetrabromobisphenol A (TBBPA) and its replacement alternative tetrabromobisphenol S (TBBPS) are used widely as brominated flame retardants (BFRs). However, the potential risk of their effects on bovine trypsin remains largely unknown. We investigated the effects of TBBPA and TBBPS to bovine trypsin by the fluorescence spectroscopy, circular dichroism and molecular dynamics (MD) simulations. They statically quenched the intrinsic fluorescence of bovine trypsin in a concentration-dependent mode and caused slight red-shifted fluorescence. The short and long fluorescence lifetime decay components of bovine trypsin were both affected, partly due to the disturbed microenvironmental changes of Trp215. The β-sheet content of bovine trypsin was significantly reduced from 82.4% to 75.7% and 76.6% by TBBPA and TBBPS, respectively, possibly impairing the physiological function of bovine trypsin. TBBPA and TBBPS bind at the 8-anilinonaphthalene-1-sulfonate (ANS) binding site with an association constant of 1.09 × 10{sup 4} M{sup −1} and 2.41 × 10{sup 4} M{sup −1} at 298 K, respectively. MD simulations revealed that van der Waals interactions and hydrogen bond interactions are dominant for TBBPA, whereas electrostatic interactions are critical for TBBPS. Our in vitro and in silico studies are beneficial to the understanding of risk assessment and future design of environmental benign BFRs.

  5. Trypsin and N-aminopeptidase (APN) activities in the hepatopancreas of an intertidal euryhaline crab: Biochemical characteristics and differential modulation by histamine and salinity.

    Science.gov (United States)

    Michiels, Maria Soledad; Del Valle, Juana Cristina; López Mañanes, Alejandra A

    2017-02-01

    No studies are available about biochemical characteristics and modulation (i.e. by endogenous and/or environmental cues) of trypsin (a key digestive endoprotease) in hepatopancreas of intertidal euryhaline crabs neither on the possible concomitant modulation of key ectoproteases such as aminopeptidase-N (APN) involved in final steps of protein digestion. Furthermore, nothing is still known in decapods crustaceans about the role of histamine as primary chemical messenger for modulation of main components of digestive process (i.e. proteases). We determined biochemical characteristics and investigated the effect of histamine injections; of histamine in vitro and of acclimation of individuals to low and high salinity on trypsin and aminopeptidase-N (APN) activities in the hepatopancreas of the euryhaline crab Cyrtograpsus angulatus (Dana 1851). Trypsin activity was maximal at pH7.4 and at 45°C. APN activity increased from pH6.6 to 7.6-9.0 and was maintained high at 37-45°C. Both activities exhibited Michaelis-Menten kinetics (apparent Km: trypsin=0.36mM; APN=0.07mM). The injection of 10-4M histamine decreased trypsin activity (about 40%) in hepatopancreas while did not affect APN activity. Similarly, in vitro 10-4M histamine decreased trypsin activity (about 52%) in hepatopancreas but not APN activity. Trypsin activity in the hepatopancreas was not affected by acclimation of crabs to low (10psu) or high (40psu) environmental salinity while APN activity was increased (about 200%) in 10psu. The results show the differential modulation of trypsin and APN by distinct cues and point to histamine as modulator of intracellular trypsin by direct action on the hepatopancreas. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. A Single Point Mutation Creating a Furin Cleavage Site in the Spike Protein Renders Porcine Epidemic Diarrhea Coronavirus Trypsin Independent for Cell Entry and Fusion.

    Science.gov (United States)

    Li, Wentao; Wicht, Oliver; van Kuppeveld, Frank J M; He, Qigai; Rottier, Peter J M; Bosch, Berend-Jan

    2015-08-01

    The emerging porcine epidemic diarrhea virus (PEDV) requires trypsin supplementation to activate its S protein for membrane fusion and virus propagation in cell culture. By substitution of a single amino acid in the S protein, we created a recombinant PEDV with an artificial furin protease cleavage site N terminal of the putative fusion peptide (PEDV-SFCS). PEDV-SFCS exhibited trypsin-independent cell-cell fusion and was able to replicate in culture cells independently of trypsin, though to low titer. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  7. Detection of viral infection by immunofluorescence in formalin-fixed tissues, pretreated with trypsin

    Directory of Open Access Journals (Sweden)

    O. M. Barth

    1988-06-01

    Full Text Available The presence of viral antigen in sections from formalin-fixed and paraffin-embedded human tissues was demonstrated by trypsin digestion followed by direct or indirect immunofluorescence. The specimens may be used for retrospective diagnosis. The immunofluorescence technique has to be adapted to the suspected virus infection on the basis of previous histopathology study. Variations of trypsin concentration time and temperature of incubation, expose different viral antigens and have to be previously tested for each unknown system. For measles virus detection in lung a stronger digestion has to be applied as compared to adenovirus or respiratory disease viruses in the same tisue. Flavivirus in liver tissue needs a weaker digestion. The reproducibility of the method makes it useful as a routine technique in diagnosis of virus infection.A presença de antígeno viral em cortes de tecidos humanos fixados em formol e emblocados em parafina foi demonstrada pela digestão com tripsina foi demonstrada pela ingestão com tripsina seguida de imunofluorescência direta ou indireta. Os espécimens podem ser utilizados para diagnoses retrospectivas. A técnica da imunofluorescência deve ser adaptada à infecção viral suspeita segundo diagnosie histopatológica prévia. Os parâmetros para a digestão do tecido pela tripsina, relacionados à concentração, duração de atuação e temperatura, expõem diferentes antígenos virais e devem ser previamente testados para cada sistema a ser estabelecido. Uma digestão mais intensa deve ser aplicada para a detecção do vírus do sarampo em tecido pulmonar do que para adenovírus ou vírus respiratório sincicial no mesmo tecido. Por outro lado, o vírus da febre amarela em tecido de fígado necessita de uma digestão mais fraca.

  8. Recognizing and Combating Sexist Job Interviews.

    Science.gov (United States)

    Garrison, Laura

    1980-01-01

    This is a self-help guide for women in occupations in which sexist treatment has occurred. The career counselor can help a woman client prepare for a job interview with the sexism-recognition skills outlined. Counselors can work with employers to lower sexist barriers. (Author/BEF)

  9. How T lymphocytes recognize lipid antigens.

    Science.gov (United States)

    De Libero, Gennaro; Mori, Lucia

    2006-10-09

    Recognition of lipid antigens by T lymphocytes is well established. Lipids are recognized by T cells when presented in association with CD1 antigen-presenting molecules. Both microbial and self lipids stimulate specific T lymphocytes, thus participating in immune reactions during infections and autoimmune diseases. The immune system uses a variety of strategies to solubilise lipid antigens, to facilitate their internalization, processing, and loading on CD1 molecules. Recent studies in the field of lipid antigen presentation have revealed new mechanisms which allow the immune system to sense lipids as stimulatory antigens.

  10. An under recognized cause of chest pain

    Directory of Open Access Journals (Sweden)

    Unnikrishnan Ponnamma Kunjan Pillai

    2012-01-01

    Full Text Available Aortic intramural hematoma (IMH is related to but is pathologically distinct from aortic dissection. In this potentially lethal entity, there is hemorrhage into the aortic media in the absence of an intimal tear. With recent advances in imaging techniques, IMH is now increasingly recognized. The limited data available suggest that the clinical course of IMH mimics that of acute aortic dissection, and mortality rates are similar. Physicians need to be cognizant regarding this entity when they are evaluating chest pain. Here we report a case of IMH, in a 63-year-old female, which was managed conservatively.

  11. The effect of trypsin digestion on the structure and iron-donating properties of transferrins from several species.

    Science.gov (United States)

    Esparza, I; Brock, J H

    1980-04-25

    The effect of trypsin digestion on iron-saturated and iron-free (apo) human, rabbit, bovine, pig and horse tranferrins has been studied. Iron-binding fragments were produced only from iron-saturated pig and bovine transferrins although some cleavage of the polypeptide chain occurred in all cases. The apo-transferrins were generally degraded to a greater extent than the corresponding iron-saturated proteins. The ability of the different transferrins to donate iron to rabbit reticulocytes varied in the order rabbit approximately pig greater than human approximately horse greater than bovine. Trypsin digestion considerably reduced the ability of pig and bovine transferrins to donate iron to rabbit reticulocytes, slightly reduced the iron-donating ability of rabbit transferrin, and had almost no effect on that of human or horse transferrins.

  12. Trypsin immobilization on an ethylenediamine-based monolithic minidisk for rapid on-line peptide mass fingerprinting studies.

    Science.gov (United States)

    Nicoli, R; Rudaz, S; Stella, C; Veuthey, J-L

    2009-03-27

    The aim of this work was to develop a trypsin-based micro-immobilized enzyme reactor prepared on a monolithic ethylenediamine BIA Separations CIM (convective interaction media) minidisk. The micro-immobilized enzyme reactor (IMER) was integrated in a liquid chromatography system hyphenated to electrospray ionization tandem mass spectrometry to carry out on-line protein digestion and identification. The performance of this IMER was compared with that obtained using a previously developed bioreactor prepared on a conventional CIM ethylenediamine disk and with that of the commercially available Poroszyme immobilized trypsin cartridge. In this work, we showed how different proteins were identified with good recoveries using a digestion time of 10 min only.

  13. TMPRSS2 and MSPL Facilitate Trypsin-Independent Porcine Epidemic Diarrhea Virus Replication in Vero Cells.

    Science.gov (United States)

    Shi, Wen; Fan, Wenlu; Bai, Jing; Tang, Yandong; Wang, Li; Jiang, Yanping; Tang, Lijie; Liu, Min; Cui, Wen; Xu, Yigang; Li, Yijing

    2017-05-18

    Type II transmembrane serine proteases (TTSPs) facilitate the spread and replication of viruses such as influenza and human coronaviruses, although it remains unclear whether TTSPs play a role in the progression of animal coronavirus infections, such as that by porcine epidemic diarrhea virus (PEDV). In this study, TTSPs including TMPRSS2, HAT, DESC1, and MSPL were tested for their ability to facilitate PEDV replication in Vero cells. Our results showed that TMPRSS2 and MSPL played significant roles in the stages of cell-cell fusion and virus-cell fusion, whereas HAT and DESC1 exhibited weaker effects. This activation may be involved in the interaction between TTSPs and the PEDV S protein, as the S protein extensively co-localized with TMPRSS2 and MSPL and could be cleaved by co-expression with TMPRSS2 or MSPL. Moreover, the use of Vero cells expressing TMPRSS2 and MSPL facilitated PEDV replication in the absence of exogenous trypsin. In sum, we identified two host proteases, TMPRSS2 and MSPL, which may provide insights and a novel method for enhancing viral titers, expanding virus production, and improving the adaptability of PEDV isolates in vitro.

  14. Three-dimensional structure of soybean trypsin/chymotrypsin Bowman-Birk inhibitor in solution

    Energy Technology Data Exchange (ETDEWEB)

    Werner, M.H.; Wemmer, D.E. (Univ. of California, Berkeley (United States))

    1992-02-04

    The three-dimensional structure of soybean trypsin/chymotrypsin Bowman-Birk inhibitor in solution has been determined by two-dimensional {sup 1}H nuclear magnetic resonance spectroscopy and dynamical simulated annealing using the program XPLOR. The structure was defined by 907 NOEs involving intra- and interresidue contacts which served as distance constraints for a protocol of dynamical simulated annealing. In addition, 48 {phi} angle constraints involving non-proline amino acids, 29 {chi} angle constraints, six {omega} angle constraints for the X-Pro peptide bond, and 35 stereoassignments for prochiral centers were incorporated during the course of the calculation. The protein is characterized by two distinct binding domains for serine protease. Each domain is comprised of a {beta}-hairpin (antiparallel {beta}-sheet and a cis-proline-containing type VIb reverse turn) with a short segment making a third strand of antiparallel {beta}-sheet. The structure determination and refinement are described, and the structure is compared to other structures of Bowman-Birk inhibitors as well as other families of serine protease inhibitors.

  15. Capsicum annuum L. trypsin inhibitor as a template scaffold for new drug development against pathogenic yeast.

    Science.gov (United States)

    Ribeiro, Suzanna F F; Silva, Marciele S; Da Cunha, Maura; Carvalho, André O; Dias, Germana B; Rabelo, Guilherme; Mello, Erica O; Santa-Catarina, Claudete; Rodrigues, Rosana; Gomes, Valdirene M

    2012-03-01

    A 6,000 Da peptide, named CaTI, was isolated from Capsicum annuum L. seeds and showed potent inhibitory activity against trypsin and chymotrypsin. The aim of this study was to determine the effect of CaTI on Saccharomyces cerevisiae, Candida albicans, Candida tropicalis and Kluyveromyces marxiannus cells. We observed that CaTI inhibited the growth of S. cerevisiae, K. marxiannus as well as C. albicans and induced cellular agglomeration and the release of cytoplasmic content. No effect on growth was observed in C. tropicalis but morphological changes were noted. In the spot assay, different degrees of sensitivity were shown among the strains and concentrations tested. Scanning electron microscopy showed that S. cerevisiae, K. marxiannus and C. albicans, in the presence of CaTI, exhibited morphological alterations, such as the formation of pseudohyphae, cellular aggregates and elongated forms. We also show that CaTI induces the generation of nitric oxide and interferes in a dose-dependent manner with glucose-stimulated acidification of the medium mediated by H(+)-ATPase of S. cerevisiae cells.

  16. Fish skin gelatin hydrolysates produced by visceral peptidase and bovine trypsin: Bioactivity and stability.

    Science.gov (United States)

    Ketnawa, Sunantha; Benjakul, Soottawat; Martínez-Alvarez, Oscar; Rawdkuen, Saroat

    2017-01-15

    The peptidase from the viscera of farmed giant catfish was used for producing gelatin hydrolysates (HG) and compared with those produced from commercial bovine trypsin (HB). The degree of hydrolysis (DH) observed suggests that proteolytic cleavage rapidly occurred within the first 120min of incubation, and there was higher DH in HG than in HB. HG demonstrated the highest ACE-inhibitory activity, DPPH, ABTS radical scavenging activity, and FRAP. HB showed the highest FRAP activity. The DPPH radical scavenging activity of HG was quite stable over the pH range of 1-11, but it increased slightly when the heating duration time reached 240min at 100°C. The ACE-inhibitory activity of HG showed the highest stability at a pH of 7, and it remained very stable at 100°C for over 15-240min. The visceral peptidase from farmed giant catfish could be an alternative protease for generating protein hydrolysates with desirable bioactivities. The resulting hydrolysates showed good stability, making them potential functional ingredients for food formulations. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Prolonged hypoxic culture and trypsinization increase the pro-angiogenic potential of human adipose tissue-derived stem cells

    DEFF Research Database (Denmark)

    Rasmussen, Jeppe Grøndahl; Frøbert, Ole; Pilgaard, Linda

    2011-01-01

    Transplantation of mesenchymal stromal cells (MSC), including adipose tissue-derived stem cells (ASC), is a promising option in the treatment of vascular disease. Short-term hypoxic culture of MSC augments secretion of anti-apoptotic and angiogenic cytokines. We hypothesized that prolonged hypoxi...... (1% and 5% oxygen) culture and trypsinization would augment ASC expression of anti-apoptotic and angiogenic cytokines and increase the angiogenic potential of ASC-conditioned media....

  18. Prolonged hypoxic culture and trypsinization increase the pro-angiogenic potential of human adipose tissue-derived stem cells

    DEFF Research Database (Denmark)

    Rasmussen, Jeppe Grøndahl; Frøbert, Ole; Pilgaard, Linda

    2011-01-01

    Transplantation of mesenchymal stromal cells (MSC), including adipose tissue-derived stem cells (ASC), is a promising option in the treatment of vascular disease. Short-term hypoxic culture of MSC augments secretion of anti-apoptotic and angiogenic cytokines. We hypothesized that prolonged hypoxic...... (1% and 5% oxygen) culture and trypsinization would augment ASC expression of anti-apoptotic and angiogenic cytokines and increase the angiogenic potential of ASC-conditioned media....

  19. Evaluation of interspecimen trypsin digestion efficiency prior to multiple reaction monitoring-based absolute protein quantification with native protein calibrators.

    Science.gov (United States)

    van den Broek, Irene; Smit, Nico P M; Romijn, Fred P H T M; van der Laarse, Arnoud; Deelder, André M; van der Burgt, Yuri E M; Cobbaert, Christa M

    2013-12-06

    Implementation of quantitative clinical chemistry proteomics (qCCP) requires targeted proteomics approaches, usually involving bottom-up multiple reaction monitoring-mass spectrometry (MRM-MS) with stable-isotope labeled standard (SIS) peptides, to move toward more accurate measurements. Two aspects of qCCP that deserve special attention are (1) proper calibration and (2) the assurance of consistent digestion. Here, we describe the evaluation of tryptic digestion efficiency by monitoring various signature peptides, missed cleavages, and modifications during proteolysis of apolipoprotein A-I and B in normo- and hypertriglyceridemic specimens. Absolute quantification of apolipoprotein A-I and B was performed by LC-MRM-MS with SIS peptide internal standards at two time points (4 and 20 h), using three native protein calibrators. Comparison with an immunoturbidimetric assay revealed recoveries of 99.4 ± 6.5% for apolipoprotein A-I and 102.6 ± 7.2% for apolipoprotein B after 4 h of trypsin digestion. Protein recoveries after 20 h trypsin incubation equaled 95.9 ± 6.9% and 106.0 ± 10.0% for apolipoproteins A-I and B, respectively. In conclusion, the use of metrologically traceable, native protein calibrators looks promising for accurate quantification of apolipoprotein A-I and B. Selection of rapidly formed peptides, that is, with no or minor missed cleavages, and the use of short trypsin incubation times for these efficiently cleaved peptides are likely to further reduce the variability introduced by trypsin digestion and to improve the traceability of test results to reach the desirable analytical performance for clinical chemistry application.

  20. Evaluation of cleaving agents other than trypsin in direct agglutination test for further improving diagnosis of visceral leishmaniasis.

    Science.gov (United States)

    el Harith, A; Chowdhury, S; al-Masum, A; Semião-Santos, S; Karim, E; el-Safi, S; Haque, I

    1995-01-01

    Trypsin treatment of Leishmania promastigote antigen has proved to be indispensible in the direct agglutination test (DAT) for the diagnosis of visceral leishmaniasis (VL) and canine visceral leishmaniasis (CVL). In the present study four antigen batches were prepared with pronase (400 micrograms/ml), lipase (0.45% [wt/vol]), pancreatin (0.3% [wt/vol]), or 2-mercaptoethanol (2-ME) (1.2% [vol/vol]) at a ratio of 20:1 versus promastigote packed cell volume or a density of 10(8)/ml. Batches prepared in this way performed satisfactorily when compared with the performance of the initial trypsinated antigen. Even higher was the sensitivity and specificity of the 2-ME-processed antigen, scoring a minimum DAT titer of 1:102,400 in the VL and CVL group and a maximum of 1:400 in the negative control group. Corresponding titers ranging from 1:6,400 to 1:12,800 and 1:800 to 1:1,600 were obtained with the antigen variants processed with pronase, lipase, pancreatin, or trypsin. By combining the use of indigenous Leishmania donovani subspecies from Sudan, Bangladesh, or Morocco and incorporating 2-ME instead of trypsin in the antigen processing step, a threefold increase in titer was attained in sera from the respective areas where VL is endemic. 2-ME-processed antigen suspensions maintained stability at 4 degrees C for up to 9 months, as evidenced by the absence of autoagglutination and the reproducibility of DAT readings with standard sera. The specificity of DAT was further improved by supplementation of the sample diluent with 0.03 M urea and incubation of the test plates at 37 degrees C for 1 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7559934

  1. Quantifying Protein-Ligand Binding Constants using Electrospray Ionization Mass Spectrometry: A Systematic Binding Affinity Study of a Series of Hydrophobically Modified Trypsin Inhibitors

    National Research Council Canada - National Science Library

    Cubrilovic, Dragana; Biela, Adam; Sielaff, Frank; Steinmetzer, Torsten; Klebe, Gerhard; Zenobi, Renato

    2012-01-01

    NanoESI-MS is used for determining binding strengths of trypsin in complex with two different series of five congeneric inhibitors, whose binding affinity in solution depends on the size of the P3 substituent...

  2. Extraction of gelatin from salmon (Salmo salar) fish skin using trypsin-aided process: optimization by Plackett-Burman and response surface methodological approaches.

    Science.gov (United States)

    Fan, HuiYin; Dumont, Marie-Josée; Simpson, Benjamin K

    2017-11-01

    Gelatin from salmon (Salmo salar) skin with high molecular weight protein chains (α-chains) was extracted using trypsin-aided process. Response surface methodology was used to optimise the extraction parameters. Yield, hydroxyproline content and protein electrophoretic profile via sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of gelatin were used as responses in the optimization study. The optimum conditions were determined as: trypsin concentration at 1.49 U/g; extraction temperature at 45 °C; and extraction time at 6 h 16 min. This response surface optimized model was significant and produced an experimental value (202.04 ± 8.64%) in good agreement with the predicted value (204.19%). Twofold higher yields of gelatin with high molecular weight protein chains were achieved in the optimized process with trypsin treatment when compared to the process without trypsin.

  3. Identification and characterization of a Bowman-Birk inhibitor active towards trypsin but not chymotrypsin in Lupinus albus seeds.

    Science.gov (United States)

    Scarafoni, Alessio; Consonni, Alessandro; Galbusera, Valerio; Negri, Armando; Tedeschi, Gabriella; Rasmussen, Patrizia; Magni, Chiara; Duranti, Marcello

    2008-06-01

    The paper describes the purification, structural characterization and inhibitory properties of a trypsin inhibitor from Lupinus albus L., a leguminous plant believed to be devoid of any protease inhibitor. The protein has been isolated by a newly set-up procedure and characterized by direct amino acid sequencing, MALDI-TOF mass spectroscopy and circular dichroism. Inhibitory properties toward bovine trypsin and chymotrypsin, as well as its thermal and pH stabilities, have been also assessed. The inhibitor is 63 amino acid long (Mr 6858; pI 8.22) and it is capable to inhibit two trypsin molecules simultaneously, with a Kd of 4.2+/-0.4 nM, but not chymotrypsin. BLAST search against UniProtKB/TrEMBL database indicates that the inhibitor belongs to the Bowman-Birk inhibitor (BBI) family. The interest in these serine-protease inhibitors arises from the ability to prevent or suppress carcinogen-induced transformation, as shown in various in vitro and in vivo model systems.

  4. In vivo efficacy of anuran trypsin inhibitory peptides against staphylococcal skin infection and the impact of peptide cyclization.

    Science.gov (United States)

    Malik, U; Silva, O N; Fensterseifer, I C M; Chan, L Y; Clark, R J; Franco, O L; Daly, N L; Craik, D J

    2015-04-01

    Staphylococcus aureus is a virulent pathogen that is responsible for a wide range of superficial and invasive infections. Its resistance to existing antimicrobial drugs is a global problem, and the development of novel antimicrobial agents is crucial. Antimicrobial peptides from natural resources offer potential as new treatments against staphylococcal infections. In the current study, we have examined the antimicrobial properties of peptides isolated from anuran skin secretions and cyclized synthetic analogues of these peptides. The structures of the peptides were elucidated by nuclear magnetic resonance (NMR) spectroscopy, revealing high structural and sequence similarity with each other and with sunflower trypsin inhibitor 1 (SFTI-1). SFTI-1 is an ultrastable cyclic peptide isolated from sunflower seeds that has subnanomolar trypsin inhibitory activity, and this scaffold offers pharmaceutically relevant characteristics. The five anuran peptides were nonhemolytic and noncytotoxic and had trypsin inhibitory activities similar to that of SFTI-1. They demonstrated weak in vitro inhibitory activities against S. aureus, but several had strong antibacterial activities against S. aureus in an in vivo murine wound infection model. pYR, an immunomodulatory peptide from Rana sevosa, was the most potent, with complete bacterial clearance at 3 mg · kg(-1). Cyclization of the peptides improved their stability but was associated with a concomitant decrease in antimicrobial activity. In summary, these anuran peptides are promising as novel therapeutic agents for treating infections from a clinically resistant pathogen. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  5. Comparison of Conventional and Microwave Treatment on Soymilk for Inactivation of Trypsin Inhibitors and In Vitro Protein Digestibility

    Directory of Open Access Journals (Sweden)

    Brinda Harish Vagadia

    2018-01-01

    Full Text Available Soymilk is lower in calories compared to cow’s milk, since it is derived from a plant source (no cholesterol and is an excellent source of protein. Despite the beneficial factors, soymilk is considered as one of the most controversial foods in the world. It contains serine protease inhibitors which lower its nutritional value and digestibility. Processing techniques for the elimination of trypsin inhibitors and lipoxygenase, which have shorter processing time and lower production costs are required for the large-scale manufacturing of soymilk. In this study, the suitable conditions of time and temperature are optimized during microwave processing to obtain soymilk with maximum digestibility with inactivation of trypsin inhibitors, in comparison to the conventional thermal treatment. The microwave processing conditions at a frequency of 2.45 GHz and temperatures of 70 °C, 85 °C and 100 °C for 2, 5 and 8 min were investigated and were compared to conventional thermal treatments at the same temperature for 10, 20 and 30 min. Response surface methodology is used to design and optimize the experimental conditions. Thermal processing was able to increase digestibility by 7% (microwave and 11% (conventional compared to control, while trypsin inhibitor activity reduced to 1% in microwave processing and 3% in conventional thermal treatment when compared to 10% in raw soybean.

  6. Functional analysis of five trypsin-like protease genes in the oriental fruit fly, Bactrocera dorsalis (Diptera: Tephritidae).

    Science.gov (United States)

    Li, Ya-Li; Hou, Ming-Zhe; Shen, Guang-Mao; Lu, Xue-Ping; Wang, Zhe; Jia, Fu-Xian; Wang, Jin-Jun; Dou, Wei

    2017-03-01

    Insect midgut proteases catalyze the release of free amino acids from dietary proteins and are essential for insect normal development. To date, digestive proteases as potential candidates have made great progress in pest control. To clarify the function of trypsin-like protease genes in the digestive system of Bactrocera dorsalis, a serious pest of a wide range of tropical and subtropical fruit and vegetable crops, five trypsin genes (BdTry1, BdTry2, BdTry3, BdTry4 and BdTry5) were identified from transcriptome dataset, and the effects of feeding condition on their expression levels were examined subsequently. RNA interference (RNAi) was applied to further explore their function on the growth of B. dorsalis. The results showed that all the BdTrys in starving midgut expressed at a minimal level but up-regulated upon feeding (except BdTry3). Besides, RNAi by feeding dsRNAs to larvae proved to be an effective method to cause gene silencing and the mixed dsRNAs of the five BdTrys slowed larvae growth of B. dorsalis. The current data suggest that trypsin genes are actively involved in digestion process of B. dorsalis larvae and thereafter play crucial roles in their development. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Flavianate, an amino acid precipitant, is a competitive inhibitor of trypsin at pH 3.0

    Directory of Open Access Journals (Sweden)

    J.M. Schneedorf

    1998-09-01

    Full Text Available Textile dyes bind to proteins leading to selective co-precipitation of a complex involving one protein molecule and more than one dye molecule of opposite charge in acid solutions, in a process of reversible denaturation that can be utilized for protein fractionation. In order to understand what occurs before the co-precipitation, a kinetic study using bovine ß-trypsin and sodium flavianate was carried out based on reaction progress curve techniques. The experiments were carried out using a-CBZ-L-Lys-p-nitrophenyl ester as substrate which was added to 50 mM sodium citrate buffer, pH 3.0, containing varying concentrations of ß-trypsin and dye. The reaction was recorded spectrophotometrically at 340 nm for 30 min, and the families of curves obtained were analyzed simultaneously by fitting integrated Michaelis-Menten equations. The dye used behaved as a competitive inhibitor of trypsin at pH 3.0, with Ki = 99 µM; kinetic parameters for the substrate hydrolysis were: Km = 32 µM, and kcat = 0.38/min. The competitive character of the inhibition suggests a specific binding of the first dye molecule to His-57, the only positively charged residue at the active site of the enzyme.

  8. Recognizing Question Entailment for Medical Question Answering.

    Science.gov (United States)

    Abacha, Asma Ben; Dina, Demner-Fushman

    2016-01-01

    With the increasing heterogeneity and specialization of medical texts, automated question answering is becoming more and more challenging. In this context, answering a given medical question by retrieving similar questions that are already answered by human experts seems to be a promising solution. In this paper, we propose a new approach for the detection of similar questions based on Recognizing Question Entailment (RQE). In particular, we consider Frequently Asked Question (FAQs) as a valuable and widespread source of information. Our final goal is to automatically provide an existing answer if FAQ similar to a consumer health question exists. We evaluate our approach using consumer health questions received by the National Library of Medicine and FAQs collected from NIH websites. Our first results are promising and suggest the feasibility of our approach as a valuable complement to classic question answering approaches.

  9. EPA recognizes industry leaders for beneficial use

    Energy Technology Data Exchange (ETDEWEB)

    Goss, D. [American Coal Ash Association (United States)

    2007-07-01

    The EPA's Coal Combustion Products Partnership C{sup 2}P{sup 2})recognized industry leaders in beneficial use during the second annual C{sup 2}P{sup 2} awards ceremony held 23 October 2006 in Atlanta, Georgia. The C{sup 2}P{sup 2} program is led by the EPA with the ACAA, DOE, FHWA, USDA - Agricultural Research Services (ARS), and Utilities Solid Waste Activities Group (USWAG). The award for overall achievement went to Great River Energy of Underwood, ND who partnered with more than 10 public and private organizations to develop an extensive market for fly ash from Coal Creek Station, the world's largest lignite-fired plant. Other awards were given for environmental achievement, innovation, partnership, research and communications and outreach. 9 photos.

  10. Fast trypsin digestion of proteins on a cross-linked [Os(dmebpy)_2Cl]^(+/2+)-derivatized copolymer of acrylamide and vinylimidazole column

    OpenAIRE

    Zhou, Jie

    2010-01-01

    Fast digestion of proteins was observed when they were loaded together with trypsin onto the crosslinked [Os(dmebpy)_2Cl]^(+/2+)-derivatized copolymer of acrylamide and vinylimidazole column. The insoluble Os-complexed polymer particles were packed into an electrospray tip to monitor peptides eluted during loading, washing and elution periods with a mass spectrometer. The proteolytic cleavage of proteins was observed immediately when the mixture of trypsin and substrates in 0.2mM ...

  11. Lol p XI, a new major grass pollen allergen, is a member of a family of soybean trypsin inhibitor-related proteins.

    Science.gov (United States)

    van Ree, R; Hoffman, D R; van Dijk, W; Brodard, V; Mahieu, K; Koeleman, C A; Grande, M; van Leeuwen, W A; Aalberse, R C

    1995-05-01

    Monoclonal antibodies were obtained against an unknown allergen from Lolium perenne grass pollen. The allergen had an apparent molecular mass of 18 kd on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Earlier immunoblotting studies had shown that carbohydrate-specific IgG antibodies recognize an antigen of similar size. We sought to characterize the allergen biochemically and immunologically. The amino acid sequence of the allergen was determined by automated Edman degradation, and its monosaccharide composition was determined by gas chromatographic analysis. A panel of 270 grass pollen-positive sera was assessed in a RAST with the purified allergen. Protease digestion (proteinase K) and chemical deglycosylation (trifluoromethane sulfonic acid) were used to distinguish between carbohydrate and peptide epitopes for IgE antibodies. The allergen was shown to be a glycoprotein with a molecular mass of 16 kd, of which 8% is carbohydrate. Its amino acid sequence shares 32% homology with soybean trypsin inhibitor (Kunitz) but lacks its active site. No homology was found with known grass pollen allergens, hence it was designated Lol p XI. A high degree of homology (44%) was found with a tree pollen allergen, Ole e I, the major allergen of olive pollen. More than 65% of grass pollen-positive sera had IgE against Lol p XI. IgE reactivity was demonstrated both with the carbohydrate moiety and the peptide backbone. Lol p XI is a new major grass pollen allergen carrying an IgE-binding carbohydrate determinant. Lol p XI is structurally related to the major allergen from olive pollen.

  12. Hydroxyl and water molecule orientations in trypsin: Comparison to molecular dynamics structures

    Energy Technology Data Exchange (ETDEWEB)

    McDowell, R.S.; Kossiakoff, A.A. [Genentech, Inc., South San Francisco, CA (United States)

    1994-12-31

    A comparison is presented of experimentally observed hydroxyl and water hydrogens in trypsin determined from neutron density maps with the results of a 140ps molecular dynamics (MD) simulation. Experimental determination of hydrogen and deuterium atom positions in molecules as large as proteins is a unique capability of neutron diffraction. The comparison addresses the degree to which a standard force-field approach can adequately describe the local electrostatic and van der Waals forces that determine the orientations of these hydrogens. Neutron densities, derived from 2.1{Angstrom} D{sub 2}O-H{sub 2}O difference Fourier maps, provide a database of 27 well-ordered hydroxyl hydrogens. Most of the simulated hydroxyl orientations are within a standard deviation of the experimentally-observed positions, including several examples in which both the simulation and the neutron density indicate that a hydroxyl group is shifted from a {open_quote}standard{close_quote} rotamer. For the most highly ordered water molecules, the hydrogen distributions calculated from the trajectory were in good agreement with neutron density; simulated water molecules that displayed multiple hydrogen bonding networks had correspondingly broadened neutron density profiles. This comparison was facilitated by development of a method to construct a pseudo 2{Angstrom} density map based on the hydrogen atom distributions from the simulation. The degree of disorder of internal water molecules is shown to result primarily from the electrostatic environment surrounding that water molecule as opposed to the cavity size available to the molecule. A method is presented for comparing the discrete observations sampled in a dynamics trajectory with the time- averaged data obtained from X-ray or neutron diffraction studies. This method is particularly useful for statically-disordered water molecules, in which the average location assigned from a trajectory may represent a site of relatively low occupancy.

  13. Allelic differentiation of Kunitz trypsin inhibitor in wild soybean (Glycine soja).

    Science.gov (United States)

    Wang, K J; Takahata, Y; Kono, Y; Kaizuma, N

    2008-08-01

    Soybean Kunitz trypsin inhibitor (SKTI) has several polymorphic types, which are controlled by co-dominant multiple alleles at a single locus. Of these types, Tia and Tib are predominant types, and there are nine differences in amino acids between Tia and Tib. Recently, an intermediate transitional type (Tibi5) between them was detected. However, other transitional types have not been detected despite surveys of many cultivated and wild soybeans. One of the reasons why other transitional variants have not been found is inferred to be due to the difficulty of the detection of SKTI protein variants by polyacrylamide gel electrophoresis (PAGE). To detect novel variants of SKTI, nucleotide sequence analysis in addition to PAGE was carried out. Four new variants were found from many Japanese wild soybeans. Of these variants, three (designated as Tiaa1, Tiaa2, Tiab1) were detected through gene sequence analysis on wild soybeans having the same electrophoretic mobility as Tia, and one (Tig) was detected through PAGE. The Tig variant showed a slightly lower electrophoretic mobility than Tic. The nucleotide sequences of Tig were identical to those of Tib except for one T-->C transitional mutation at position +340. The sequences of Tiaa1 and Tiaa2 genes were identical to those of Tia with the exception of a G-->A mutation at position +376 and a T-->C mutation at +404, respectively. The sequence of Tiab1 differed from Tia by three nucleotides: C-->A at position +331, T-->C at +459 and A-->G at +484. Of the three nucleotide changes, two were common to Tiab1, Tibi5 and Tib, suggesting that Tiab1 is an intermediate transitional type between Tia and Tib. Our results suggest that Tib type has been differentiated through a series of mutations from Tia before the domestication of cultivated soybean.

  14. Protein surface softness is the origin of enzyme cold-adaptation of trypsin.

    Directory of Open Access Journals (Sweden)

    Geir Villy Isaksen

    2014-08-01

    Full Text Available Life has effectively colonized most of our planet and extremophilic organisms require specialized enzymes to survive under harsh conditions. Cold-loving organisms (psychrophiles express heat-labile enzymes that possess a high specific activity and catalytic efficiency at low temperatures. A remarkable universal characteristic of cold-active enzymes is that they show a reduction both in activation enthalpy and entropy, compared to mesophilic orthologs, which makes their reaction rates less sensitive to falling temperature. Despite significant efforts since the early 1970s, the important question of the origin of this effect still largely remains unanswered. Here we use cold- and warm-active trypsins as model systems to investigate the temperature dependence of the reaction rates with extensive molecular dynamics free energy simulations. The calculations quantitatively reproduce the catalytic rates of the two enzymes and further yield high-precision Arrhenius plots, which show the characteristic trends in activation enthalpy and entropy. Detailed structural analysis indicates that the relationship between these parameters and the 3D structure is reflected by significantly different internal protein energy changes during the reaction. The origin of this effect is not localized to the active site, but is found in the outer regions of the protein, where the cold-active enzyme has a higher degree of softness. Several structural mechanisms for softening the protein surface are identified, together with key mutations responsible for this effect. Our simulations further show that single point-mutations can significantly affect the thermodynamic activation parameters, indicating how these can be optimized by evolution.

  15. {sup 13}C-NMR studies on disulfide bond isomerization in bovine pancreatic trypsin inhibitor (BPTI)

    Energy Technology Data Exchange (ETDEWEB)

    Takeda, Mitsuhiro [Kumamoto University, Department of Structural BioImaging, Faculty of Life Sciences (Japan); Miyanoiri, Yohei [Nagoya University, Structural Biology Research Center, Graduate School of Science (Japan); Terauchi, Tsutomu [Tokyo Metropolitan University, Graduate School of Science and Engineering (Japan); Kainosho, Masatsune, E-mail: kainosho@tmu.ac.jp [Nagoya University, Structural Biology Research Center, Graduate School of Science (Japan)

    2016-09-15

    Conformational isomerization of disulfide bonds is associated with the dynamics and thus the functional aspects of proteins. However, our understanding of the isomerization is limited by experimental difficulties in probing it. We explored the disulfide conformational isomerization of the Cys14–Cys38 disulfide bond in bovine pancreatic trypsin inhibitor (BPTI), by performing an NMR line-shape analysis of its Cys carbon peaks. In this approach, 1D {sup 13}C spectra were recorded at small temperature intervals for BPTI samples selectively labeled with site-specifically {sup 13}C-enriched Cys, and the recorded peaks were displayed in the order of the temperature after the spectral scales were normalized to a carbon peak. Over the profile of the line-shape, exchange broadening that altered with temperature was manifested for the carbon peaks of Cys14 and Cys38. The Cys14–Cys38 disulfide bond reportedly exists in equilibrium between a high-populated (M) and two low-populated states (m{sub c14} and m{sub c38}). Consistent with the three-site exchange model, biphasic exchange broadening arising from the two processes was observed for the peak of the Cys14 α-carbon. As the exchange broadening is maximized when the exchange rate equals the chemical shift difference in Hz between equilibrating sites, semi-quantitative information that was useful for establishing conditions for {sup 13}C relaxation dispersion experiments was obtained through the carbon line-shape profile. With respect to the m{sub c38} isomerization, the {sup 1}H-{sup 13}C signals at the β-position of the minor state were resolved from the major peaks and detected by exchange experiments at a low temperature.

  16. Effects of organic solvents and substrate binding on trypsin in acetonitrile and hexane media.

    Science.gov (United States)

    Meng, Yanyan; Yuan, Yuan; Zhu, Yanyan; Guo, Yanzhi; Li, Menglong; Wang, Zhimeng; Pu, Xuemei; Jiang, Lin

    2013-09-01

    In this work, we used molecular dynamic (MD) simulation to study trypsin with and without a six-amino-acid peptide bound in three different solvents (water, acetonitrile and hexane) in order to provide molecular information for well understanding the structure and function of enzymes in non-aqueous media. The results show that the enzyme is more compact and less native-like in hexane than in the other two polar solvents. The substrate could stabilize the native protein structure in the two polar media, but not in the non-polar hexane. There are no significant differences in the conformation of the S1 pocket upon the substrate binding in water and acetonitrile media while a reverse behavior is observed in hexane media, implying a possible induced fit binding mechanism in the non-polar media. The substrate binding enhances the stability of catalytic H-bond network since it could expel the solvent molecules from the active site. The enzyme and the substrate appear to be more appropriate to the reactive conformation in the organic solvents compared with aqueous solution. There is much greater substrate binding strength in hexane media than the water and acetonitrile ones since the polar solvent significantly weakens electrostatic interactions, which are observed to be the main driving force to the binding. In addition, some residues of the S1 pocket could remain favorable contribution to the binding despite the solvent change, but with differences in the contribution extent, the number and the type of residues between the three media.

  17. Arabic word recognizer for mobile applications

    Science.gov (United States)

    Khanna, Nitin; Abdollahian, Golnaz; Brame, Ben; Boutin, Mireille; Delp, Edward J.

    2011-03-01

    When traveling in a region where the local language is not written using a "Roman alphabet," translating written text (e.g., documents, road signs, or placards) is a particularly difficult problem since the text cannot be easily entered into a translation device or searched using a dictionary. To address this problem, we are developing the "Rosetta Phone," a handheld device (e.g., PDA or mobile telephone) capable of acquiring an image of the text, locating the region (word) of interest within the image, and producing both an audio and a visual English interpretation of the text. This paper presents a system targeted for interpreting words written in Arabic script. The goal of this work is to develop an autonomous, segmentation-free Arabic phrase recognizer, with computational complexity low enough to deploy on a mobile device. A prototype of the proposed system has been deployed on an iPhone with a suitable user interface. The system was tested on a number of noisy images, in addition to the images acquired from the iPhone's camera. It identifies Arabic words or phrases by extracting appropriate features and assigning "codewords" to each word or phrase. On a dictionary of 5,000 words, the system uniquely mapped (word-image to codeword) 99.9% of the words. The system has a 82% recognition accuracy on images of words captured using the iPhone's built-in camera.

  18. Overview: recognizing the problem of magnesium deficiency

    Energy Technology Data Exchange (ETDEWEB)

    Seelig, M.S.

    1988-01-01

    The magnesium content of the usual American diet is less than the recommended dietary allowance. Excesses of some macro- and micro-nutrients interact with Mg, increasing its requirements. Marginal deficiency of Mg is not associated with hypomagnesemia, is not characterized by typical manifestations, as is thus difficult to diagnose. Serum or plasma Mg levels are held within narrow limits unless tissue levels are very low, or renal function is poor. Vulnerability to Mg deficiency increases during growth and development, pregnancy, when under physical or psychological stress, and during illness or its treatment that interferes with absorption or causes loss of Mg. Evidence of biochemical changes of early Mg deficiency is rarely sought, although the roles of Mg in many enzyme systems are recognized. The effects of Mg deficiency on metabolism, even in disorders caused by vitamin dependencies in which Mg is a co-factor, are largely unexplored. Deficiency of Mg is diagnosed confidently when the laboratory reports hypomagnesemia in patients with convulsions or arrhythmias. Without these signs, Mg levels are not often ordered, even in the presence of neuromuscular irritability such as respond to Mg repletion. Because Mg supplementation or Mg-sparing drugs protect against premature or ectopic heart beats and sudden death, to which diuretic-treated hypertensive patients are at risk, it is increasingly being advised that their Mg status be determined.

  19. Perspective: Recognizing and rewarding clinical scholarship.

    Science.gov (United States)

    Grigsby, R Kevin; Thorndyke, Luanne

    2011-01-01

    Faculty members in medical schools and academic medical centers are in a constant process of generating new knowledge. The cornerstone of academia--and academic medicine--is scholarship. Traditionally, tenure and/or academic promotion in the professorial ranks is awarded to those who meet institutional criteria in the missions of research, teaching, and service, including patient care. In the academic review process, priority is often placed on a record of demonstrated, consistent success in traditional laboratory research, also known as the scholarship of discovery. More recently, there has been greater recognition of other forms of scholarship: education, application, and integration. These forms of scholarship, although less recognized, also result in the generation of new knowledge. In an attempt to understand the breadth and scope of clinical scholarship, the authors searched the extant literature in academic medicine for a definition of clinical scholarship and expanded the search to disciplines outside of medicine. They found that succinct, discrete definitions of clinical scholarship have been published in other disciplines, but not in academic medicine. After reviewing definitions of clinical scholarship from other disciplines, adapting definitions of educational scholarship in academic medicine, and including qualities unique to clinical scholarship, the authors developed a framework for understanding clinical scholarship in academic medicine as a means for opening a dialogue within the academic medical community. This dialogue hopefully will lead to formulating a succinct, discrete definition of clinical scholarship that will allow greater recognition and reward for clinical scholars in the promotion and tenure process.

  20. Escherichia coli fimbriae recognizing sialyl galactosides.

    Science.gov (United States)

    Korhonen, T K; Väisänen-Rhen, V; Rhen, M; Pere, A; Parkkinen, J; Finne, J

    1984-08-01

    Fimbriae recognizing sialyl galactosides (S fimbriae) were purified from an Escherichia coli strain. The S fimbriae were morphologically identical to type 1 and P fimbriae of E. coli and showed a hemagglutination that was abolished when erythrocytes were treated with neuraminidase. Hemagglutination by the purified fimbriae was inhibited by orosomucoid but not by its desialylated derivative. Of the oligosaccharides tested, sialyl-(alpha 2-3)-lactose and sialyl-(alpha 2-3)-N-acetyllactosamine had the strongest inhibitory activities. It was concluded that S fimbriae have the strongest affinity for (alpha 2-3)-linked sialyl galactosides. In the enzyme-linked immunosorbent assay, the hyperimmune serum to the S fimbriae reacted strongly with the homologous antigen but not with type 1, P, or nonhemagglutinating KS71C fimbriae of E. coli. Analogously, the hyperimmune sera to the other E. coli fimbriae did not react with the purified S fimbriae. The immunoprecipitation assay showed that S fimbriae on different E. coli serotypes shared immunological cross-reactivity.

  1. Dogs recognize dog and human emotions

    Science.gov (United States)

    Albuquerque, Natalia; Wilkinson, Anna; Savalli, Carine; Otta, Emma; Mills, Daniel

    2016-01-01

    The perception of emotional expressions allows animals to evaluate the social intentions and motivations of each other. This usually takes place within species; however, in the case of domestic dogs, it might be advantageous to recognize the emotions of humans as well as other dogs. In this sense, the combination of visual and auditory cues to categorize others' emotions facilitates the information processing and indicates high-level cognitive representations. Using a cross-modal preferential looking paradigm, we presented dogs with either human or dog faces with different emotional valences (happy/playful versus angry/aggressive) paired with a single vocalization from the same individual with either a positive or negative valence or Brownian noise. Dogs looked significantly longer at the face whose expression was congruent to the valence of vocalization, for both conspecifics and heterospecifics, an ability previously known only in humans. These results demonstrate that dogs can extract and integrate bimodal sensory emotional information, and discriminate between positive and negative emotions from both humans and dogs. PMID:26763220

  2. Dogs recognize dog and human emotions.

    Science.gov (United States)

    Albuquerque, Natalia; Guo, Kun; Wilkinson, Anna; Savalli, Carine; Otta, Emma; Mills, Daniel

    2016-01-01

    The perception of emotional expressions allows animals to evaluate the social intentions and motivations of each other. This usually takes place within species; however, in the case of domestic dogs, it might be advantageous to recognize the emotions of humans as well as other dogs. In this sense, the combination of visual and auditory cues to categorize others' emotions facilitates the information processing and indicates high-level cognitive representations. Using a cross-modal preferential looking paradigm, we presented dogs with either human or dog faces with different emotional valences (happy/playful versus angry/aggressive) paired with a single vocalization from the same individual with either a positive or negative valence or Brownian noise. Dogs looked significantly longer at the face whose expression was congruent to the valence of vocalization, for both conspecifics and heterospecifics, an ability previously known only in humans. These results demonstrate that dogs can extract and integrate bimodal sensory emotional information, and discriminate between positive and negative emotions from both humans and dogs. © 2016 The Author(s).

  3. Platelets (thrombocytes: The other recognized functions

    Directory of Open Access Journals (Sweden)

    Nikolić Ljubinka I.

    2016-01-01

    Full Text Available Introduction: Platelets are smallest blood cells of discoid or round shape and are cytoplasmic fragments of megakaryocytes. Platelets consist of 3 types of granules: alpha granules, dense granules and lysosomes. Granule secretion releases coagulation factors, growth factors, cytokines, and a number of proteolytic enzymes. Platelets contain a number of receptors known as platelet agonists. Basic and most studied role of platelets is in hemostasis process. The aim of this paper is to point on platelet function unrelated to hemostasis. Matherials and methods: Papers on other recognized functions of platelets were searched for in biomedical journals idexed in MEDLINE form 2004 to 2016. Topic: This paper studies less known platelet functions becoming subject of interest with the development of applied science. Platelets participate in inflammation by releasing proinflammatory mediators (CD154, CD40L. Complement activation via Pselectin, platelet-generates immunomodulatory effect. CD40L accelerates releasing RANTES protein leading to intensified activation of T-lymphocytes. During embryonic development, platelets allow blood and lymph vessels separation by activating CLE-2 receptor and ligand PDPN. Platelets alleviate migration and invasiveness of tumor cells, contribute to disease progression and development of metastases. Platelets affect maturation of follicles and oocytes and have important role in embryo implantation process and placentation. Conclusion: Based on these findings, conclusion imposes platelets as not only active participants in the hemostasis process but as having significant role in inflammation, unspecified and specified body defending, tumor biology, embryonic development and in female reproductive system regulation. Numerous roles of platelets open wide range for the new drugs' operation. Their specific characteristic is the basis for the personalized Clinical pharmacology development and possibility of applying specific drug as

  4. Can a CNN recognize Catalan diet?

    Science.gov (United States)

    Herruzo, P.; Bolaños, M.; Radeva, P.

    2016-10-01

    Nowadays, we can find several diseases related to the unhealthy diet habits of the population, such as diabetes, obesity, anemia, bulimia and anorexia. In many cases, these diseases are related to the food consumption of people. Mediterranean diet is scientifically known as a healthy diet that helps to prevent many metabolic diseases. In particular, our work focuses on the recognition of Mediterranean food and dishes. The development of this methodology would allow to analise the daily habits of users with wearable cameras, within the topic of lifelogging. By using automatic mechanisms we could build an objective tool for the analysis of the patient's behavior, allowing specialists to discover unhealthy food patterns and understand the user's lifestyle. With the aim to automatically recognize a complete diet, we introduce a challenging multi-labeled dataset related to Mediter-ranean diet called FoodCAT. The first type of label provided consists of 115 food classes with an average of 400 images per dish, and the second one consists of 12 food categories with an average of 3800 pictures per class. This dataset will serve as a basis for the development of automatic diet recognition. In this context, deep learning and more specifically, Convolutional Neural Networks (CNNs), currently are state-of-the-art methods for automatic food recognition. In our work, we compare several architectures for image classification, with the purpose of diet recognition. Applying the best model for recognising food categories, we achieve a top-1 accuracy of 72.29%, and top-5 of 97.07%. In a complete diet recognition of dishes from Mediterranean diet, enlarged with the Food-101 dataset for international dishes recognition, we achieve a top-1 accuracy of 68.07%, and top-5 of 89.53%, for a total of 115+101 food classes.

  5. Thermodynamic evaluation and modeling of proton and water exchange associated with benzamidine and berenil binding to ß-trypsin

    Directory of Open Access Journals (Sweden)

    M.T. Pereira

    2005-11-01

    Full Text Available Serine-proteases are involved in vital processes in virtually all species. They are important targets for researchers studying the relationships between protein structure and activity, for the rational design of new pharmaceuticals. Trypsin was used as a model to assess a possible differential contribution of hydration water to the binding of two synthetic inhibitors. Thermodynamic parameters for the association of bovine ß-trypsin (homogeneous material, observed 23,294.4 ± 0.2 Da, theoretical 23,292.5 Da with the inhibitors benzamidine and berenil at pH 8.0, 25ºC and with 25 mM CaCl2, were determined using isothermal titration calorimetry and the osmotic stress method. The association constant for berenil was about 12 times higher compared to the one for benzamidine (binding constants are K = 596,599 ± 25,057 and 49,513 ± 2,732 M-1, respectively; the number of binding sites is the same for both ligands, N = 0.99 ± 0.05. Apparently the driving force responsible for this large difference of affinity is not due to hydrophobic interactions because the variation in heat capacity (DCp, a characteristic signature of these interactions, was similar in both systems tested (-464.7 ± 23.9 and -477.1 ± 86.8 J K-1 mol-1 for berenil and benzamidine, respectively. The results also indicated that the enzyme has a net gain of about 21 water molecules regardless of the inhibitor tested. It was shown that the difference in affinity could be due to a larger number of interactions between berenil and the enzyme based on computational modeling. The data support the view that pharmaceuticals derived from benzamidine that enable hydrogen bond formation outside the catalytic binding pocket of ß-trypsin may result in more effective inhibitors.

  6. Pre-diagnostic levels of anionic trypsinogen, cationic trypsinogen, and pancreatic secretory trypsin inhibitor in relation to pancreatic cancer risk.

    Science.gov (United States)

    Johansen, Dorthe; Manjer, Jonas; Regner, Sara; Lindkvist, Björn

    2010-01-01

    Experimental studies have suggested that trypsinogen may enhance tumor progression and that the ratio between anionic trypsinogen and cationic trypsinogen (HAT/HCT) and between the sum of trypsinogens and pancreatic secretory trypsin inhibitor (PSTI) ((HAT + HCT)/PSTI) are disturbed in patients with pancreatic cancer. The aim of this study was to investigate if pre-diagnostic levels of these parameters are associated with subsequent pancreatic cancer risk. A total of 33,346 subjects participated in a health screening programme in Malmö, Sweden. Pancreatic cancer cases (n = 84) were matched to three controls each. HAT, HCT and PSTI were analyzed in pre-diagnostic serum samples. Odds ratios for pancreatic cancer were calculated using logistic regression and were then stratified for other risk factors. In the main analysis, a statistically significant association between the ratio between HAT/HCT and pancreatic cancer was observed for all, for the crude OR and for the ORs adjusted for sex, BMI or Helicobacter pylori. When stratified for sex, statistically significant associations were found for females in the crude OR and for the ORs adjusted for time to analysis, BMI, alcohol consumption or H. pylori. There was a positive association between the ratio of HAT/HCT to pancreatic cancer in the intermediate/high alcohol consumption group and subjects with a BMI alcohol consumption group. Our hypothesis predicted an increased risk for pancreatic cancer related to an imbalance between trypsin activity and trypsin inhibition capacity. The findings concerning the ratio of HAT/HCT are in line with this. The results related to analyses stratified for other risk factors should be considered as mainly explorative. Copyright 2010 S. Karger AG, Basel.

  7. Proteolytic profiling and comparative analyses of active trypsin-like serine peptidases in preimaginal stages of Culex quinquefasciatus

    Directory of Open Access Journals (Sweden)

    Borges-Veloso Andre

    2012-06-01

    Full Text Available Abstract Background The mosquito Culex quinquefasciatu s, a widespread insect in tropical and sub-tropical regions of the world, is a vector of multiple arboviruses and parasites, and is considered an important risk to human and veterinary health. Proteolytic enzymes play crucial roles in the insect physiology including the modulation of embryonic development and food digestion. Therefore, these enzymes represent important targets for the development of new control strategies. This study presents zymographic characterization and comparative analysis of the proteolytic activity found in eggs, larval instars and pupae of Culex quinquefasciatus. Methods The proteolytic profiles of eggs, larvae and pupa of Cx. quinquefasciatus were characterized by SDS-PAGE co-polymerized with 0.1% gelatin, according to the pH, temperature and peptidase inhibitor sensitivity. In addition, the proteolytic activities were characterized in solution using 100 μM of the fluorogenic substrate Z-Phe-Arg-AMC. Results Comparison of the proteolytic profiles by substrate-SDS-PAGE from all preimaginal stages of the insect revealed qualitative and quantitative differences in the peptidase expression among eggs, larvae and pupae. Use of specific inhibitors revealed that the proteolytic activity from preimaginal stages is mostly due to trypsin-like serine peptidases that display optimal activity at alkaline pH. In-solution, proteolytic assays of the four larval instars using the fluorogenic substrate Z-Phe-Arg-AMC in the presence or absence of a trypsin-like serine peptidase inhibitor confirmed the results obtained by substrate-SDS-PAGE analysis. The trypsin-like serine peptidases of the four larval instars were functional over a wide range of temperatures, showing activities at 25°C and 65°C, with an optimal activity between 37°C and 50°C. Conclusion The combined use of zymography and in-solution assays, as performed in this study, allowed for a more detailed analysis of the

  8. Allium sativum Protease Inhibitor: A Novel Kunitz Trypsin Inhibitor from Garlic Is a New Comrade of the Serpin Family.

    Directory of Open Access Journals (Sweden)

    Tooba Naz Shamsi

    Full Text Available This study was aimed to purify and characterize the Protease inhibitor (PI from a plant Allium sativum (garlic with strong medicinal properties and to explore its phytodrug potentials.Allium sativum Protease Inhibitor (ASPI was purified using ammonium sulphate fractionation and Fast Protein Liquid Chromatography on anion exchanger Hi-Trap DEAE column. The purified protein was analyzed for its purity and molecular weight by SDS PAGE. The confirmation of presence of trypsin inhibiting PI was performed by MALDI TOF-TOF and analyzed by MASCOT database. The ASPI was further investigated for its kinetic properties and stability under extreme conditions of pH, temperature and chemical denaturants. Secondary structure was determined by Circular Dichorism (CD spectroscopy.ASPI of ~15 kDa inhibited trypsin and matched "truncated kunitz Trypsin Inhibitor (Glycine max" in MASCOT database. The purified ASPI showed 30376.1371 U/mg specific activity with a fold purity of 159.92 and yield ~93%. ASPI was quite stable in the range of pH 2-12 showing a decline in the activity around pH 4-5 suggesting that the pI value of the protein as ASPI aggregates in this range. ASPI showed stability to a broad range of temperature (10-80°C but declined beyond 80°C. Further, detergents, oxidizing agents and reducing agents demonstrated change in ASPI activity under varying concentrations. The kinetic analysis revealed sigmoidal relationship of velocity with substrate concentration with Vmax 240.8 (μM/min and Km value of 0.12 μM. ASPI showed uncompetitive inhibition with a Ki of 0.08±0.01 nM. The Far UV CD depicted 2.0% α -helices and 51% β -sheets at native pH.To conclude, purified ~15 kDa ASPI exhibited fair stability in wide range of pH and temperature Overall, there was an increase in purification fold with remarkable yield. Chemical modification studies suggested the presence of lysine and tryptophan residues as lead amino acids present in the reactive sites

  9. Allium sativum Protease Inhibitor: A Novel Kunitz Trypsin Inhibitor from Garlic Is a New Comrade of the Serpin Family.

    Science.gov (United States)

    Shamsi, Tooba Naz; Parveen, Romana; Amir, Mohd; Baig, Mohd Affan; Qureshi, M Irfan; Ali, Sher; Fatima, Sadaf

    2016-01-01

    This study was aimed to purify and characterize the Protease inhibitor (PI) from a plant Allium sativum (garlic) with strong medicinal properties and to explore its phytodrug potentials. Allium sativum Protease Inhibitor (ASPI) was purified using ammonium sulphate fractionation and Fast Protein Liquid Chromatography on anion exchanger Hi-Trap DEAE column. The purified protein was analyzed for its purity and molecular weight by SDS PAGE. The confirmation of presence of trypsin inhibiting PI was performed by MALDI TOF-TOF and analyzed by MASCOT database. The ASPI was further investigated for its kinetic properties and stability under extreme conditions of pH, temperature and chemical denaturants. Secondary structure was determined by Circular Dichorism (CD) spectroscopy. ASPI of ~15 kDa inhibited trypsin and matched "truncated kunitz Trypsin Inhibitor (Glycine max)" in MASCOT database. The purified ASPI showed 30376.1371 U/mg specific activity with a fold purity of 159.92 and yield ~93%. ASPI was quite stable in the range of pH 2-12 showing a decline in the activity around pH 4-5 suggesting that the pI value of the protein as ASPI aggregates in this range. ASPI showed stability to a broad range of temperature (10-80°C) but declined beyond 80°C. Further, detergents, oxidizing agents and reducing agents demonstrated change in ASPI activity under varying concentrations. The kinetic analysis revealed sigmoidal relationship of velocity with substrate concentration with Vmax 240.8 (μM/min) and Km value of 0.12 μM. ASPI showed uncompetitive inhibition with a Ki of 0.08±0.01 nM). The Far UV CD depicted 2.0% α -helices and 51% β -sheets at native pH. To conclude, purified ~15 kDa ASPI exhibited fair stability in wide range of pH and temperature Overall, there was an increase in purification fold with remarkable yield. Chemical modification studies suggested the presence of lysine and tryptophan residues as lead amino acids present in the reactive sites. Therefore, ASPI

  10. Solvent exchange of buried water and hydrogen exchange of peptide NH groups hydrogen bonded to buried waters in bovine pancreatic trypsin inhibitor

    Energy Technology Data Exchange (ETDEWEB)

    Tuechsen, E.; Hayes, J.M.; Ramaprasad, S.; Copie, V.; Woodward, C.

    1987-08-11

    Solvent exchange of /sup 18/O-labeled buried water in bovine pancreatic trypsin inhibitor (BPTI), trypsin, and trypsin-BPTI complex is measured by high-precision isotope ratio mass spectroscopy. Buried water is labeled by equilibrium of the protein in /sup 18/O-enriched water. Protein samples are then rapidly dialyzed against water of normal isotope composition by gel filtration and stored. The exchangeable /sup 18/O label eluting with the protein in 10-300 s is determined by an H/sub 2/O-CO/sub 2/ equilibration technique. Exchange of buried waters with solvent water is complete before 10-15 s in BPTI, trypsin, and BPTI-trypsin, as well as in lysozyme and carboxypeptidase measured as controls. When in-exchange dialysis and storage are carried out at pH greater than or equal to 2.5, trypsin-BPTI and trypsin, but not free BPTI, have the equivalent of one /sup 18/O atom that exchanges slowly (after 300 s and before several days). This oxygen is probably covalently bound to a specific site in trypsin. When in-exchange dialysis and storage are carried out at pH 1.1, the equivalent of three to seven /sup 18/O atoms per molecule is associated with the trypsin-BPTI complex, apparently due to nonspecific covalent /sup 18/O labeling of carboxyl groups at low pH. In addition to /sup 18/O exchange of buried waters, the hydrogen isotope exchange of buried NH groups H bonded to buried waters was also measured. Their base-catalyzed exchange rate constants are on the order of NH groups that in the crystal are exposed to solvent and hydrogen-bonded main chain O, and their pH/sub min/ is similar to that for model compounds. The pH dependence of their exchange rate constants suggests that direct exchange with water may significantly contribute to their observed exchange rate.

  11. Conformational flexibility in the catalytic triad revealed by the high-resolution crystal structure of Streptomyces erythraeus trypsin in an unliganded state

    Energy Technology Data Exchange (ETDEWEB)

    Blankenship, Elise; Vukoti, Krishna [Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106 (United States); Miyagi, Masaru, E-mail: mxm356@cwru.edu [Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106 (United States); Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106 (United States); Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106 (United States); Lodowski, David T., E-mail: mxm356@cwru.edu [Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106 (United States); Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106 (United States)

    2014-03-01

    This work reports the first sub-angstrom resolution structure of S. erythraeus trypsin. The detailed model of a prototypical serine protease at a catalytically relevant pH with an unoccupied active site is presented and is compared with other high-resolution serine protease structures. With more than 500 crystal structures determined, serine proteases make up greater than one-third of all proteases structurally examined to date, making them among the best biochemically and structurally characterized enzymes. Despite the numerous crystallographic and biochemical studies of trypsin and related serine proteases, there are still considerable shortcomings in the understanding of their catalytic mechanism. Streptomyces erythraeus trypsin (SET) does not exhibit autolysis and crystallizes readily at physiological pH; hence, it is well suited for structural studies aimed at extending the understanding of the catalytic mechanism of serine proteases. While X-ray crystallographic structures of this enzyme have been reported, no coordinates have ever been made available in the Protein Data Bank. Based on this, and observations on the extreme stability and unique properties of this particular trypsin, it was decided to crystallize it and determine its structure. Here, the first sub-angstrom resolution structure of an unmodified, unliganded trypsin crystallized at physiological pH is reported. Detailed structural analysis reveals the geometry and structural rigidity of the catalytic triad in the unoccupied active site and comparison to related serine proteases provides a context for interpretation of biochemical studies of catalytic mechanism and activity.

  12. Urine of patients with early prostate cancer contains lower levels of light chain fragments of inter-alpha-trypsin inhibitor and saposin B but increased expression of an inter-alpha-trypsin inhibitor heavy chain 4 fragment.

    Science.gov (United States)

    Jayapalan, Jaime J; Ng, Keng L; Shuib, Adawiyah S; Razack, Azad H A; Hashim, Onn H

    2013-06-01

    The present study was aimed at the identification of proteins that are differentially expressed in the urine of patients with prostate cancer (PCa), those with benign prostatic hyperplasia (BPH) and age-matched healthy male control subjects. Using a combination of 2DE and MS/MS, significantly lower expression of urinary saposin B and two different fragments of inter-alpha-trypsin inhibitor light chain (ITIL) was demonstrated in the PCa patients compared to the controls. However, only one of the ITIL fragments was significantly different between the PCa and BPH patients. When image analysis was performed on urinary proteins that were transferred onto NC membranes and detected using a lectin that binds to O-glycans, a truncated fragment of inter-alpha-trypsin inhibitor heavy chain 4 was the sole protein found to be significantly enhanced in the PCa patients compared to the controls. Together, these urinary peptide fragments might be useful complementary biomarkers to indicate PCa as well as to distinguish it from BPH, although further epidemiological evidence on the specificity and sensitivity of the protein candidates is required. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Lifting as We Climb: Recognizing Intersectional Gender Violence in Law

    Directory of Open Access Journals (Sweden)

    Shreya Atrey

    2015-12-01

    Full Text Available This paper interrogates the meaning of lifting all women as we climb the ladder of gender equality and justice by recognizing that gender violence affects women differently. This is because violence against women is perpetrated not only on the basis of their gender or sex but also other identities of race, religion, caste, region, age, disability, nationality, sexual orientation etc. With reference to CEDAW jurisprudence and examples from India, I seek to explain this understanding with the help of a normative framework of ‘intersectional integrity’. The framework insists on considering claimants as a whole by tracing unique and shared patterns of gender violence when it is also based on other identities such as race, religion, caste, region, age, disability, nationality, and sexual orientation. I argue that applying the framework allows us to diagnose and address the nature of violence suffered on multiple identities, in a clear and comprehensive way. Este artículo cuestiona el sentido de levantar a todas las mujeres a medida que se asciende la escalera de la igualdad de género y la justicia, reconociendo que la violencia de género afecta a las mujeres de manera diferente. Esto se debe a que la violencia contra las mujeres se comete no sólo sobre la base de su género o sexo, sino también por su raza, religión, casta, región, edad, discapacidad, nacionalidad, orientación sexual, etc. Se pretende explicar esta afirmación con la ayuda de un marco normativo de “integridad interseccional”, a través de referencias a la jurisprudencia del CEDAW y ejemplos de la India. El marco insiste en considerar a las demandantes en su conjunto, trazando patrones únicos y compartidos de violencia de género cuando se basa también en otras identidades como raza, religión, casta, región, edad, discapacidad, nacionalidad, orientación sexual. Se sostiene que la aplicación del marco permite diagnosticar y abordar la naturaleza de la violencia

  14. Heat-induced inactivation mechanisms of Kunitz trypsin inhibitor and Bowman-Birk inhibitor in soymilk processing.

    Science.gov (United States)

    Chen, Yeming; Xu, Zhicun; Zhang, Caimeng; Kong, Xiangzhen; Hua, Yufei

    2014-07-01

    Trypsin inhibitor activity (TIA) is an important antinutritional factor in soymilk. In this study, the effects of NaCl preaddition on TIA and the heat-induced TIA inactivation mechanisms were examined. The results showed that Kunitz trypsin inhibitor (KTI) and Bowman-Birk inhibitor (BBI) contributed 74% and 26% to raw soymilk TIA, respectively. The heat-induced quick KTI incorporation into protein aggregates was the reason for its quick TIA inactivation. The heat-induced slow cleavage of one BBI peptide bond was the reason for its slow TIA inactivation. Heat-induced protein aggregate formation had little effect on BBI inactivation owing to the fact that BBI and its degradation product tended to remain in the supernatant (197,000g, 1h) in all conditions used in this study. NaCl could accelerate the KTI incorporation into protein aggregates and the cleavage of one BBI peptide bond, which supplied a simple and quick method for low TIA soymilk processing. Copyright © 2013 Elsevier Ltd. All rights reserved.

  15. Serine proteinase from Cucurbita ficifolia seed; purification, properties, substrate specificity and action on native squash trypsin inhibitor (CMTI I).

    Science.gov (United States)

    Dryjanski, M; Otlewski, J; Polanowski, A; Wilusz, T

    1990-09-01

    A proteinase was purified from resting seeds of Cucurbita ficifolia by ammonium sulfate fractionation and successive chromatography on CM-cellulose, Sephacryl S-300 and TSK DEAE-2SW (HPLC) columns. Inhibition by DFP and PMSF suggests that the enzyme is a serine proteinase. The apparent molecular mass of this enzyme is ca. 77 kDa. The optimum activity for hydrolysis of casein and Suc-Ala-Ala-Pro-Phe-pNA is around pH 10.5. The following peptide bonds in the oxidized insulin B-chain were hydrolysed by the proteinase: Phe1-Val2, Asn3-Gln4, Gln4-His5, Cya7-Gly8, Glu13-Ala14, Ala14-Leu15, Cya19-Gly20, Pro28-Lys29 and Lys29-Ala30. The proteinase is more selective towards the native squash seed trypsin inhibitor (CMTI I) and primarily cuts off only its N-terminal arginine. The inhibitor devoided of the N-terminal arginine residue is still active against trypsin.

  16. Purification and characterisation of trypsin-like enzyme from the pyloric caeca of cod (Gadus morhua II

    Directory of Open Access Journals (Sweden)

    Luiz Henrique Beirão

    2001-03-01

    Full Text Available A trypsin -like enzyme from the pyloric caeca of cod (Gadus morhua was purified by affinity chromatography on CHOM Sepharose 4B. Some characteristics were established by its catalytic activity on T.A.M.E., typical enzyme substrate, and serine protease inhibitors. The enzyme had an isoelectric point of 5.30 and 5.89 and was very similar in amino acid composition to bovine trypsin, but differed in having a higher relative amount of acidic amino acids and a lower amount of basic amino acids. The enzyme also hydrolysed fish protein substrates.A tripsina do ceco pilórico do bacalhau (Gadus morhua foi purificada por cromatografia de CHOM Sepharose 4B. Algumas características foram determinadas como atividade catalítica para T.A.M.E., substratos enzimáticos e inibidores de proteases. A enzima mostrou ponto isoelétrico de 5,30 e 5,89 e composição de aminoácidos similar à tripsina bovina, mas diferiu por ter um alto percentual de aminoácidos ácidos e baixo valor em aminoácidos básicos. A tripsina do bacalhau também hidrolisou substratos de proteínas de pescados.

  17. Pancreatic mass, cellularity, and alpha-amylase and trypsin activity in feedlot steers fed diets differing in crude protein concentration.

    Science.gov (United States)

    Swanson, K C; Kelly, N; Salim, H; Wang, Y J; Holligan, S; Fan, M Z; McBride, B W

    2008-04-01

    Twenty-four yearling beef steers (initial BW = 510 +/- 4.9 kg) predominantly of Angus breeding were used in a randomized complete block design to determine the effect of dietary CP concentration on pancreatic cellularity, mass, and alpha-amylase and trypsin activities. Treatment diets were formulated to contain 8.8, 11.0, 13.2, and 15.4% CP. Soybean meal and Top Soy (ruminal bypass soybean meal) were used as supplemental protein sources to ensure that MP intake was increased with increasing dietary CP concentrations. Steers were penned in groups of 4 (1 steer per treatment) and individually fed at 2.5x the NE(m) requirement by using Calan gates for 28 d before tissue collection. Four steers (1 pen) were slaughtered per week. Pancreases were weighed, subsampled, frozen in liquid N(2), and stored at -80 degrees C until analyses for DNA, RNA, and protein concentrations, and alpha-amylase and trypsin activities. Pancreatic weight (g and g/kg of BW) did not differ among treatment groups. Pancreatic DNA concentration (mg/g) decreased linearly (P = 0.06) with increasing CP concentration. Pancreatic protein (g/pancreas) increased linearly (P = 0.08) with increasing dietary CP concentration. Pancreatic alpha-amylase activity (U/g, U/mg of DNA, U/g of protein, U/pancreas, and U/kg of BW) increased linearly (P pancreas, and U/kg of BW) increased linearly (P pancreas of cattle.

  18. Cystic fibrosis screening in neonates--measurement of immunoreactive trypsin and direct genotype analysis for delta F508 mutation.

    Science.gov (United States)

    Larsen, J; Campbell, S; Faragher, E B; Götz, M; Eichler, I; Waldherr, S; Dobianer, K; Spona, J

    1994-08-01

    This study investigated the clinical usefulness of screening for cystic fibrosis (CF) in 19,992 newborns, over 39 months, in an Austrian population. Immunoreactive serum trypsin (IRT) determination was followed by sweat chloride analysis (sweat test) to establish diagnosis. In a retrospective analysis covering 6 months of the study period, individuals who were considered to be at risk after IRT estimation (n = 22) were analysed for delta F508 mutation, using a new method of DNA extraction from the initial dried blood specimens. A total of 119 infants (0.6%) had values greater than 750 ng trypsin/ml whole blood. In 88 babies sweat tests were performed, leading to the diagnosis of CF in 11 cases. One patient was not initially identified by screening but was later discovered due to his clinical status. Three infants were noted to carry the delta F508 mutation (1 homozygous, 2 heterozygous). Two of these babies already had CF. The second heterozygote was a carrier. A highly efficient three tier screening strategy is presented in which IRT estimation, determination of delta F508 status and sweat chloride testing could lead to a high sensitivity analysis of this population.

  19. Immunoreactivity and trypsin sensitivity of recombinant virus-like particles of foot-and-mouth disease virus.

    Science.gov (United States)

    Basagoudanavar, S H; Hosamani, M; Tamil, R P; Sreenivasa, B P; Chandrasekhar, B K; Venkataramanan, R

    2015-03-01

    Foot-and-mouth disease (FMD) is an important infection affecting the health and productivity of cloven-hoofed livestock. Development of improved vaccines and diagnostic reagents is being explored to facilitate the disease control. There is an emerging interest in virus-like particles (VLPs), as their constituent structural proteins are the major immunogens. The VLPs are similar to natural virus particles but lack viral nucleic acid. The objective of the present study was to express the VLPs of FMD virus (FMDV) serotype Asia-1 (IND 63/72), using baculovirus system and characterize them for antigenic structure. The VLPs expressed in insect cells showed immunoreactivity similar to inactivated cell culture FMDV. Further they possess similar sensitivity to trypsin as the inactivated cell culture FMDV, suggesting that trypsin-sensitive antigenic sites could be similarly arranged. Our findings suggest that the FMD VLPs have similar antigenic conformational feature like the wild type virus, thus supporting their utility in development of non-infectious FMD vaccines and/or diagnostic assays.

  20. Effect of hemoglobin powder substituted for fishmeal on growth performance, protein digestibility, and trypsin gene expression in Litopenaeus vannamei

    Directory of Open Access Journals (Sweden)

    Duangrat Chookird

    2010-05-01

    Full Text Available Recent increases in the price and demand of fishmeal, the primary protein source in shrimp feeds, have caused asearch for alternative protein sources. Hemoglobin powder (HE is a by-product produced by separating hemoglobin fromplasma of farm animal un-coagulant blood. HE contains high protein content but low lipid content, and thus has highpotential for fishmeal substitution.A six week feeding trial was carried out to investigate effects of HE substituted for fishmeal protein on growth performance,protein digestibility and trypsin gene expression in Pacific white shrimp (Litopenaeus vannamei. Six diets with0%, 12.5%, 25%, 50%, 75% and 100% of HE replacing fishmeal protein were fed four times daily to six groups of shrimp withan average initial weight of 3.53 g/shrimp. Growth of shrimp decreased with increasing level of HE substitution. Although the12.5% HE substitution caused significantly lower final weight, weight gain, SGR, feed intake, PER and PPV in comparisonwith the control diet, FCR of this diet was not statistically different (p0.05. In-vitro and in-vivo protein digestibility of 12.5% HE substitution were significantly lower than that of the controlgroup in which the trypsin gene expression of shrimp fed 12.5% HE substituted diet was the highest.

  1. Isolation and Characterization of a Phaseolus vulgaris Trypsin Inhibitor with Antiproliferative Activity on Leukemia and Lymphoma Cells.

    Science.gov (United States)

    Li, Miao; Liu, Qin; Cui, Yajuan; Li, Dong; Wang, Hexiang; Ng, Tzi Bun

    2017-01-23

    A 17.5-kDa trypsin inhibitor was purified from Phaseolus vulgaris cv. "gold bean" with an isolation protocol including ion exchange chromatography on DEAE-cellulose (Diethylaminoethyl-cellulose), affinity chromatography on Affi-gel blue gel, ion exchange chromatography on SP-sepharose (Sulfopropyl-sepharose), and gel filtration by FPLC (Fast protein liquid chromatography) on Superdex 75. It dose-dependently inhibited trypsin with an IC50 value of 0.4 μM, and this activity was reduced in the presence of dithiothreitol in a dose- and time-dependent manner, signifying the importance of the disulfide linkage to the activity. It inhibited [methyl-³H] thymidine incorporation by leukemia L1210 cells and lymphoma MBL2 cells with an IC50 value of 2.3 μM and 2.5 μM, respectively. The inhibitor had no effect on fungal growth and the activities of various viral enzymes when tested up to 100 μM.

  2. Microchip bioreactors based on trypsin-immobilized graphene oxide-poly(urea-formaldehyde) composite coating for efficient peptide mapping.

    Science.gov (United States)

    Fan, Huizhi; Yao, Feina; Xu, Shuyuan; Chen, Gang

    2013-12-15

    Trypsin was covalently immobilized to graphene oxide (GO)-poly(urea-formaldehyde) (PUF) composite coated on the channel wall of poly(methyl methacrylate) microchips to fabricate microfluidic bioreactors for highly efficient proteolysis. A mixture solution containing urea-formaldehyde prepolymer and GO nanosheets was allowed to flow through the channels. The modification layer on the channel wall could further polycondense to form GO-PUF composite coating in the presence of ammonium chloride. The primary amino groups of trypsin could react with the carboxyl groups of the GO sheets in the coating with the aid of carboxyl activating agents to realize covalent immobilization. The feasibility and performance of the novel GO-based microchip bioreactors were demonstrated by the digestion of bovine serum albumin, lysozyme, ovalbumin, and myoglobin. The digestion time was significantly reduced to less than 5s. The obtained digests were identified by MALDI-TOF MS with satisfactory sequence coverages that were comparable to those obtained by using 12-h in-solution digestion. The present proteolysis strategy is simple and efficient, offering great promise for high-throughput protein identification. © 2013 Elsevier B.V. All rights reserved.

  3. Re-discovering Women's Leadership Potential through Improved ...

    African Journals Online (AJOL)

    . However, only few churches recognized this and make use of women. It is against this background that this paper highlights the hidden potentials of women and benefits that the church could derived from women leadership using Deeper life ...

  4. Diabulimia: what it is and how to recognize it in critical care.

    Science.gov (United States)

    Ruth-Sahd, Lisa A; Schneider, Melissa; Haagen, Brigitte

    2009-01-01

    Critical care nurses must be able to recognize the signs of symptoms of diabulimia-a potentially life-threatening disorder. Skipping insulin is used as a means of weight control in some persons with diabetes, particularly in young women. This article focuses on the assessment, pathophysiology, critical care nursing interventions, and psychosocial initiatives of interest to critical care nurses in the care of patients with diabulimia.

  5. Fast trypsin digestion of proteins on a cross-linked [Os(dmebpy)(2)Cl](+/2+)-derivatized copolymer of acrylamide and vinylimidazole column.

    Science.gov (United States)

    Zhou, Jie

    2010-08-15

    Fast digestion of proteins was observed when they were loaded together with trypsin onto the cross-linked [Os(dmebpy)(2)Cl](+/2+)-derivatized copolymer of acrylamide and vinylimidazole column. The insoluble Os-complexed polymer particles were packed into an electrospray tip to monitor peptides eluted during loading, washing and elution periods with a mass spectrometer. The proteolytic cleavage of proteins was observed immediately when the mixture of trypsin and substrates in 0.2 mM ammonium bicarbonate 50:50 H(2)O/acetonitrile reached the column tip, and continued through the loading period. Some tryptic peptides were released from the column during the loading and following washing periods. The others still stayed on the column until the low pH elution buffer reached the column. If a protein was first loaded onto the column, no tryptic peptides of the protein were observed when trypsin was loaded later for the on-column digestion. Only the autolysis peptides of trypsin were observed. On-column digestion of 100 fmol myoglobin was successfully detected with a low sensitivity quadrupole mass spectrometer. A hybrid Os-polymer/C(18) column tip was constructed for the online trypsin digestion of proteins in the aqueous buffers and the following trapping and elution of peptides from the C(18) column. The digestion of reduced and alkylated bovine serum albumin and human transferrin in 2.5 mM ammonium bicarbonate and 0.2 M urea buffer was observed on the column, with more peptide coverage than conventional 4 h in-solution digestion at 37 degrees C. Control experiments without the Os-polymer in the column tip excluded the spontaneous in-solution digestion of proteins in the short time window of buffer delivery onto the column, indirectly confirming the contribution of Os-polymer on the fast trypsin digestion. Copyright (c) 2010 John Wiley & Sons, Ltd.

  6. Women, criminality and multifocal empowerment responses: some ...

    African Journals Online (AJOL)

    Although the overall agenda of the women's movement is committed to recognizing the needs and aspirations of women and ending gender-based inequality, issues concerning women and crime have always been ignored. this paper documents the situation of women in prison in Botswana. although the proportion of ...

  7. Recognizing Bedside Events Using Thermal and Ultrasonic Readings

    Directory of Open Access Journals (Sweden)

    Danielsen Asbjørn

    2017-06-01

    Full Text Available Falls in homes of the elderly, in residential care facilities and in hospitals commonly occur in close proximity to the bed. Most approaches for recognizing falls use cameras, which challenge privacy, or sensor devices attached to the bed or the body to recognize bedside events and bedside falls. We use data collected from a ceiling mounted 80 × 60 thermal array combined with an ultrasonic sensor device. This approach makes it possible to monitor activity while preserving privacy in a non-intrusive manner. We evaluate three different approaches towards recognizing location and posture of an individual. Bedside events are recognized using a 10-second floating image rule/filter-based approach, recognizing bedside falls with 98.62% accuracy. Bed-entry and exit events are recognized with 98.66% and 96.73% accuracy, respectively.

  8. Preliminary studies on the effect of rebamipide against the trypsin and egg-albumin induced experimental model of asthma.

    Science.gov (United States)

    Gohil, Priyanshee; Thakkar, Himani; Gohil, Unnati; Deshpande, Shrikalp

    2011-12-01

    The present investigation was carried out to study the effect of rebamipide in experimentally induced bronchial asthma in mice. Trypsin and egg-albumin induced chronic model of asthma was used and various parameters were measured on the 35th day. The asthmatic control group showed lower level of haemoglobin saturation with oxygen, tidal volume, airflow rate and higher respiratory rate, serum bicarbonate level, eosinophil count in bronchoalveolar lavage fluid and histamine level compared to the normal control group. Dexamethasone and rebamipide treated groups showed the return of all the above parameters towards normal values. Histopathological examination of lungs showed more prominent alveolar and muscular layer destruction in the asthmatic control group than in dexamethasone and rebamipide treated groups. Rebamipide showed a beneficial effect and might be used for the treatment of bronchial asthma.

  9. [Soluble high molecular weight derivatives of trypsin pancreatic inhibitor. Isolation and properties of dextran-bound pancreatic inhibitor].

    Science.gov (United States)

    Larionova, N I; Kazanskaia, N F; Sakharov, I Iu

    1977-07-01

    A method of isolating preparations of pancreatic inhibitor of trypsin, bound with soluble polysaccharide carriers, is worked out. It is demonstrated that the reaction of a pancreatic inhibitor and cyanuric chloride-activated dextran proceeds for OH groups of tyrosine residues and for-epsilon-NH2 groups of lysine residues. A method is offered of the protection of amino groups with citraconic anhydride for the complete retaining of the inhibitory activity during attachment to dextran. Thermic denaturation of pancreatic inhibitor preparations at pH 4.7 and 97 degrees C is studied. It is found that the modification by 2-amino-4.6-dichloro-s-triazine stabilizes the protein molecule, while the interaction with the matrix of soluble dextran does not carry any contribution to thermostability of the pancreatic inhibitor.

  10. [Use of soybean trypsin inhibitor for modification of gold surface of the sensor chips in surface plasmon resonance spectrometer].

    Science.gov (United States)

    Avilov, S V; Aleksandrova, N O; Kunda, Ie M; Verevka, S V; Shyrshov, Iu M

    2004-01-01

    Kunitz soybean trypsin inhibitor (STI) and glutaraldehyde are used to create an interlayer for proteins immobilization on the gold surface of the sensor chips of surface plasmon resonance spectrometer. Human serum albumin (HSA), goat Ig G and bovine eye lens alpha-crystallin are immobilized via the proposed interlayer. We studied the effects of the duration of storage of the sensor chips before use and pre-treatment by "piranha" solution on the STI adsorption by the gold surface. The influence of STI surface concentration, as well as the effect of the duration of storage of STI-modified sensor chips on the HSA immobilization are investigated. The binding of specific antibodies to the immobilized proteins and non-specific binding to the modified surfaces are studied. HSA immobilization on the bare gold surface is compared to that on the surfaces, modified by different methods.

  11. Active 1918 pandemic flu viral neuraminidase has distinct N-glycan profile and is resistant to trypsin digestion.

    Science.gov (United States)

    Wu, Zhengliang L; Ethen, Cheryl; Hickey, Gregg E; Jiang, Weiping

    2009-02-13

    The 1918 pandemic flu virus caused one of the most deadly pandemics in human history. To search for unique structural features of the neuraminidase from this virus that might have contributed to its unusual virulence, we expressed this enzyme. The purified enzyme appeared as a monomer, a dimer and a tetramer, with only the tetramer being active and therefore biologically relevant. The monomer and the dimer could not be oligomerized into the tetramer in solution, suggesting that some unique structural features were required for oligomerization and activation. These features could be related to N-glycosylation, because the tetramer displayed different N-glycans than the monomer and the dimer. Furthermore, the tetramer was found to be resistant to trypsin digestion, which may give the virus the capability to invade tissues that are normally not infected by influenza viruses and make the virus more robust for infection.

  12. The control of hyperglycemia by a novel trypsin resistant oral insulin preparation in alloxan induced type I diabetic mice.

    Science.gov (United States)

    Bank, Sarbashri; Ghosh, Arjun; Bhattacharya, Suman; Maiti, Smarajit; Khan, Gausal A; Sinha, Asru K

    2016-05-26

    A trypsin resistant oral insulin preparation was made by incubating insulin for 2 h at 23 °C with previously boiled cow milk at 100 °C that was coagulated with 0.6 M acetic acid. The precipitate was resuspended in the same volume of milk. The immunoblot analysis of the suspended proteins treated with 200 ng of trypsin/ml for 3 h demonstrated that the 80.1% of the insulin in the suspension survived the proteolytic degradation compared to 0% of the hormone survived in the control. The feeding of 0.4 ml (0.08 unit of insulin) of the resuspended proteins followed by 0.2 ml of the same protein to alloxan induced diabetic mice maximally decreased the blood glucose level from 508 ± 10 mg/dl to 130 ± 10 mg/dl in 7 h with simultaneous increase of the basal plasma concentration of insulin from 3 ± 1.1 μunits/ml to 18 ± 1.5 μunits/ml. In control experiment the absence of insulin in the identical milk suspension produced no hypoglycemic effect suggesting milk was not responsible for the hypoglycemic effect of milk-insulin complex. Coming out of insulin-casein complex from the intestinal gut to the circulation was spontaneous and facilitated diffusion transportation which was found from Gibbs free energy reaction.

  13. Systematic Design of Trypsin Cleavage Site Mutated Exendin4-Cysteine 1, an Orally Bioavailable Glucagon-Like Peptide-1 Receptor Agonist

    Directory of Open Access Journals (Sweden)

    Wenbo Sai

    2017-03-01

    Full Text Available Exendin-4 is a strong therapeutic candidate for the treatment of metabolic syndrome. Related receptor agonist drugs have been on the market since 2005. However, technical limitations and the pain caused by subcutaneous injection have severely limited patient compliance. The goal of the study is to investigate a biologically active exendin-4 analog could be administered orally. Using intraperitoneal glucose tolerance tests, we discovered that exendin4-cysteine administered by oral gavage had a distinct hypoglycemic effect in C57BL/6J mice. Using Rosetta Design and Amber, we designed and screened a series of exendin4-cysteine analogs to identify those that retained biological activity while resisting trypsin digestion. Trypsin Cleavage Site Mutated Exendin4-cysteine 1 (TSME-1, an analog whose bioactivity was similar to exendin-4 and was almost completely resistant to trypsin, was screened out. In addition, TSME-1 significantly normalized the blood glucose levels and the availability of TSME-1 was significantly higher than that of exendin-4 and exendin4-cysteine. Collectively orally administered TSME-1, a trypsin-resistant exendin-4 analog obtained by the system, is a strong candidate for future treatments of type 2 diabetes.

  14. Effect of traditional processing methods on the β-carotene, ascorbic acid and trypsin inhibitor content of orange-fleshed sweet potato for production of amala in Nigeria.

    Science.gov (United States)

    Yusuf, Abbas Bazata; Fuchs, Richard; Nicolaides, Linda

    2016-05-01

    The aim of the work was to study the effect of traditional processing methods on the β-carotene, ascorbic acid and trypsin inhibitor contents of orange-fleshed sweet potato amala. The most common sweet potato in Nigeria is white or yellow fleshed, which is very low in provitamin A. However, efforts are underway to promote orange-fleshed sweet potato to improve provitamin A intake. This paper describes how orange-fleshed sweet potato slices were traditionally processed into amala, which is increasingly consumed in Nigeria. The study revealed that both the cold and hot fermentation methods resulted in increased vitamin A levels and lower vitamin C levels in orange-fleshed sweet potato. Further processing to make amala resulted in a fall in both vitamin A and C content. The study found an increase in trypsin inhibitor activity following the cold-water fermentation and a decrease following the hot-water fermentation compared to raw orange-fleshed sweet potato. Trypsin inhibitor activity in amala produced using both the cold and hot methods was below detectable levels. The results indicate that amala produced from traditionally fermented orange-fleshed sweet potato could be a good source of vitamins A and C for the rural poor and that the processing removes any potential negative effects of trypsin inhibitors. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  15. Trypsin inhibitors from the garden four o'clock (Mirabilis jalapa) and spinach (Spinacia oleracea) seeds: isolation, characterization and chemical synthesis.

    Science.gov (United States)

    Kowalska, Jolanta; Pszczoła, Katarzyna; Wilimowska-Pelc, Anna; Lorenc-Kubis, Irena; Zuziak, Ewa; Ługowski, Mateusz; Łegowska, Anna; Kwiatkowska, Anna; Sleszyńska, Małgorzata; Lesner, Adam; Walewska, Aleksandra; Zabłotna, Ewa; Rolka, Krzysztof; Wilusz, Tadeusz

    2007-06-01

    Five serine proteinase inhibitors (Mirabilis jalapa trypsin inhibitors, MJTI I and II and Spinacia oleracea trypsin inhibitors, SOTI I, II, and III) from the garden four-o'clock (M. jalapa) and spinach (S. oleracea) seeds were isolated. The purification procedures included affinity chromatography on immobilized methylchymotrypsin in the presence of 5M NaCl, ion exchange chromatography and/or preparative electrophoresis, and finally RP-HPLC on a C-18 column. The inhibitors, crosslinked by three disulfide bridges, are built of 35 to 37 amino-acid residues. Their primary structures differ from those of known trypsin inhibitors, but showed significant similarity to the antimicrobial peptides isolated from the seeds of M. jalapa (MJ-AMP1, MJ-AMP2), Mesembryanthemum crystallinum (AMP1), and Phytolacca americana (AMP-2 and PAFP-S) and from the hemolymph of Acrocinus longimanus (Alo-1, 2 and 3). The association equilibrium constants (K(a)) with bovine beta-trypsin for the inhibitors from M. jalapa (MJTI I and II) and S. oleracea (SOTI I-III) were found to be about 10(7)M(-1). Fully active MJTI I and SOTI I were obtained by solid-phase peptide synthesis. The disulfide bridge pattern in both inhibitors (Cys1-Cys4, Cys2-Cys5 and Cys3-Cys6) was established after their digestion with thermolysin and proteinase K followed by the MALDI-TOF analysis.

  16. Cell-surface mucosubstances from trypsin disaggregation of normal and virus-transformed lines of baby-hamster kidney cells (Short Communication)

    Science.gov (United States)

    Minnikin, S. Megan; Allen, Adrian

    1973-01-01

    Cell disaggregation by trypsin solubilizes significantly less mucosubstance from the surface of polyoma-virus-transformed baby-hamster kidney cells than from the same non-transformed cell line. The mucosubstance, which consists of both acid mucopolysaccharides and mucoproteins, also differs qualitatively in the two cell lines. PMID:4357713

  17. Two Kunitz-type inhibitors with activity against trypsin and papain from Pithecellobium dumosum seeds: purification, characterization, and activity towards pest insect digestive enzyme.

    Science.gov (United States)

    Oliveira, A S; Migliolo, L; Aquino, R O; Ribeiro, J K C; Macedo, L L P; Bemquerer, M P; Santos, E A; Kiyota, S; de Sales, M P

    2009-01-01

    Two trypsin inhibitors (called PdKI-3.1 and PdKI-3.2) were purified from the seeds of the Pithecellobium dumosum tree. Inhibitors were obtained by TCA precipitation, affinity chromatography on Trypsin-Sepharose and reversed-phase-HPLC. SDS-PAGE analysis with or without reducing agent showed that they are a single polypeptide chain, and MALDI-TOF analysis determined molecular masses of 19696.96 and 19696.36 Da, respectively. The N-terminal sequence of both inhibitors showed strong identity to the Kunitz family trypsin inhibitors. They were stable over a wide pH (2-9) and temperature (37 to 100 degrees C) range. These inhibitors reduced over 84% of trypsin activity with inhibition constant (Ki) of 4.20 x 10(-8) and 2.88 x 10(-8) M, and also moderately inhibited papain activity, a cysteine proteinase. PdKI-3.1 and PdKI-3.2 mainly inhibited digestive enzymes from Plodia interpunctella, Zabrotes subfasciatus and Ceratitis capitata guts. Results show that both inhibitors are members of the Kunitz-inhibitor family and that they affect the digestive enzyme larvae of diverse orders, indicating a potential insect antifeedant.

  18. Understanding binding affinity : A combined isothermal titration calorimetry/molecular dynamics study of the binding of a series of hydrophobically modified benzamidinium chloride inhibitors to trypsin

    NARCIS (Netherlands)

    Talhout, Reinskje; Villa, Alessandra; Mark, AE; Engberts, JBFN

    2003-01-01

    The binding of a series of p-alkylbenzamidinium chloride inhibitors to the serine proteinase trypsin over a range of temperatures has been studied using isothermal titration (micro)calorimetry and molecular dynamics simulation techniques. The inhibitors have small structural variations at the para

  19. TsAg5, a Taenia solium cysticercus protein with a marginal trypsin-like activity in the diagnosis of human neurocysticercosis

    Science.gov (United States)

    Rueda, Analiz; Sifuentes, Cecilia; Gilman, Robert H.; Gutiérrez, Andrés H.; Piña, Ruby; Chile, Nancy; Carrasco, Sebastián; Larson, Sandra; Mayta, Holger; Verástegui, Manuela; Rodriguez, Silvia; Gutiérrez-Correa, Marcel; García, Héctor H.; Sheen, Patricia; Zimic, Mirko

    2011-01-01

    Neurocysticercosis is an endemic parasitic disease caused by Taenia solium larva. Although the mechanism of infection is not completely understood, it is likely driven by proteolytic activity that degrades the intestinal wall to facilitate oncosphere penetration and further infection. We analyzed the publicly available Taenia solium EST/DNA library and identified two contigs comprising a full-length cDNA fragment very similar to E. granulosus Ag5 protein. The Taenia solium cDNA sequence included a proteolytic trypsin-like-domain in the C-terminal region, and a thrombospondin type-1 adherence-domain in the N-terminal region. Both the trypsin-like and adherence domains were expressed independently as recombinant proteins in bacterial systems. TsAg5 showed marginal trypsin-like activity and high sequence similarity to Ag5. The purified antigens were tested in a Western immunoblot assay to diagnose human neurocysticercosis. The sensitivity of the trypsin-like-domain was 96.36% in patients infected with extraparenchymal cysts, 75.44% in patients infected with multiple cysts, and 39.62% in patients with a single cyst. Specificity was 76.70%. The thrombospondin type-1 adherence-domain was not specific for neurocysticercosis. PMID:21893105

  20. Collagenolytic serine protease PC and trypsin PC from king crab Paralithodes camtschaticus: cDNA cloning and primary structure of the enzymes

    Directory of Open Access Journals (Sweden)

    Rebrikov Denis V

    2004-01-01

    Full Text Available Abstract Background In this paper, we describe cDNA cloning of a new anionic trypsin and a collagenolytic serine protease from king crab Paralithodes camtschaticus and the elucidation of their primary structures. Constructing the phylogenetic tree of these enzymes was undertaken in order to prove the evolutionary relationship between them. Results The mature trypsin PC and collagenolytic protease PC contain 237 (Mcalc 24.8 kDa and 226 amino acid residues (Mcalc 23.5 kDa, respectively. Alignments of their amino acid sequences revealed a high degree of the trypsin PC identity to the trypsin from Penaeus vannamei (approximately 70% and of the collagenolytic protease PC identity to the collagenase from fiddler crab Uca pugilator (76%. The phylogenetic tree of these enzymes was constructed. Conclusions Primary structures of the two mature enzymes from P. camtschaticus were obtained and compared with those of other proteolytic proteins, including some enzymes from brachyurans. A phylogenetic analysis was also carried out. These comparisons revealed that brachyurins are closely related to their vertebrate and bacterial congeners, occupy an intermediate position between them, and their study significantly contributes to the understanding of the evolution and function of serine proteases.

  1. Identification of a new soybean Kunitz trypsin inhibitor mutation and its effect on Bowman-Birk protease inhibitor content in soybean seed

    Science.gov (United States)

    Soybean seeds possess anti-nutritional compounds which inactivate digestive proteases, principally corresponding to two families: Kunitz Trypsin Inhibitors (KTi) and Bowman-Birk Inhibitors (BBI). High levels of raw soybeans/soybean meal in feed mixtures can cause poor weight gain and pancreatic abno...

  2. Women and sustainable development.

    Science.gov (United States)

    Meena, R

    1992-07-01

    Gender issues in sustainable development focuses on constraints, the policy environment, land rights, the division of labor, reproductive rights, human resource development, productive energy, care of children, education, politics, security, social norms, and women's initiatives. African women's participation in the development process has been limited by the policy environment, sociocultural setting, and women's initiatives. African policy has not recognized the different roles that men and women play. There is unequal division of labor, legal discrimination against women, and abuse of women's basic human rights. Women's subordinate position in society and their concrete needs are ignored. Land tenure and credit systems are based on discriminatory policies. Women share a major portion and in some cases all of the agricultural labor with few tools or equipment. The operating assumption is that women's labor supply is inelastic. In order to fully participate in the development process, women need to be able to determine the number of children needed, the spacing between children, and the timing and the method of contraception. Human resource development in Africa has focused on training men. Women must contribute a major portion of time and labor to processing and cooking food in addition to caring for children. Access to higher education is limited. Political accords have been reached without women when women have contributed significantly to political struggles. Social security is compromised during violence and civil strife. There is sexual harassment in the work place. Culture can subordinate women. Women have been unable to change policy making, planning, and patriarchal ideology. Women are marginal contributors to the labor force. Income-generating projects are primarily confined to the informal sector. The governments impose the women's programs. Political influence is highly desired if change in women's stature is to be accomplished.

  3. Recognizing Multi-user Activities using Body Sensor Networks

    DEFF Research Database (Denmark)

    Gu, Tao; Wang, Liang; Chen, Hanhua

    2011-01-01

    activity classes of data—for building activity models and design a scalable, noise-resistant, Emerging Pattern based Multi-user Activity Recognizer (epMAR) to recognize both single- and multi-user activities. We develop a multi-modal, wireless body sensor network for collecting real-world traces in a smart...

  4. Recognizing Strokes in Tennis Videos Using Hidden Markov Models

    NARCIS (Netherlands)

    Petkovic, M.; Jonker, Willem; Zivkovic, Z.

    This paper addresses content-based video retrieval with an emphasis on recognizing events in tennis game videos. In particular, we aim at recognizing different classes of tennis strokes using automatic learning capability of Hidden Markov Models. Driven by our domain knowledge, a robust player

  5. A novel dianionic amino acid ionic liquid-coated PEG 4000 modified Fe3O4 nanocomposite for the magnetic solid-phase extraction of trypsin.

    Science.gov (United States)

    Yang, Qin; Wang, Yuzhi; Zhang, Hongmei; Xu, Kaijia; Wei, Xiaoxiao; Xu, Panli; Zhou, Yigang

    2017-11-01

    A novel magnetic extractant, PEG 4000 modified Fe3O4nanomaterial that coated with dianionic amino acid ionic liquid (Fe3O4@PEG@DAAAIL), was successfully synthesized and characterized. X-ray diffraction (XRD), transmission electron microscope (TEM), vibrating sample magnetometer (VSM), fourier transform infrared spectrometry (FT-IR), thermal gravimetric analysis (TGA) and zeta potentials were used to confirm that the novel nanocomposite was successfully synthesized. Subsequently, the prepared Fe3O4@PEG@DAAAIL nanocomposite was used as the extractant for trypsin coupled with magnetic solid-phase extraction (MSPE). The concentrations of trypsin in the supernatant were detected by UV-vis spectrophotometer at 278nm. The extraction ability turned out to be better than the other four kinds of extractants prepared in this work. Furthermore, the influence of a series of factors, such as extraction time and temperature, initial trypsin concentration, the value of pH and ionic strength, was systematically investigated. Under the optimal extraction condition, the extraction capacity for trypsin could reach up to 718.73mg/g, absolutely higher than that of other adsorbents reported. This satisfactory extraction capacity could be maintained unchangeable after at least eight days, and kept over 90% of initial extraction capacity after eight recycles. What's more, the activity of trypsin after extraction retained 92.29% of initial activity, verifying the biocompatibility of the prepared extractant. Finally, the developed Fe3O4@PEG@DAAAIL-MSPE method was successfully applied to the real sample analysis with satisfactory results. All of above proves the potential value of Fe3O4@PEG@DAAAIL-MSPE in the analysis of biomass. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Trypsin immobilization on hairy polymer chains hybrid magnetic nanoparticles for ultra fast, highly efficient proteome digestion, facile 18O labeling and absolute protein quantification.

    Science.gov (United States)

    Qin, Weijie; Song, Zifeng; Fan, Chao; Zhang, Wanjun; Cai, Yun; Zhang, Yangjun; Qian, Xiaohong

    2012-04-03

    In recent years, quantitative proteomic research attracts great attention because of the urgent needs in biological and clinical research, such as biomarker discovery and verification. Currently, mass spectrometry (MS) based bottom up strategy has become the method of choice for proteomic quantification. In this strategy, the amount of proteins is determined by quantifying the corresponding proteolytic peptides of the proteins, therefore highly efficient and complete protein digestion is crucial for achieving accurate quantification results. However, the digestion efficiency and completeness obtained using conventional free protease digestion is not satisfactory for highly complex proteomic samples. In this work, we developed a new type of immobilized trypsin using hairy noncross-linked polymer chains hybrid magnetic nanoparticle as the matrix aiming at ultra fast, highly efficient proteomic digestion and facile (18)O labeling for absolution protein quantification. The hybrid nanoparticle is synthesized by in situ growth of hairy polymer chains from the magnetic nanoparticle surface using surface initiated atom transfer radical polymerization technique. The flexible noncross-linked polymer chains not only provide large amount of binding sites but also work as scaffolds to support three-dimensional trypsin immobilization which leads to increased loading amount and improved accessibility of the immobilized trypsin. For complex proteomic samples, obviously increased digestion efficiency and completeness was demonstrated by 27.2% and 40.8% increase in the number of identified proteins and peptides as well as remarkably reduced undigested proteins residues compared with that obtained using conventional free trypsin digestion. The successful application in absolute protein quantification of enolase from Thermoanaerobacter tengcongensis protein extracts using (18)O labeling and MRM strategy further demonstrated the potential of this hybrid nanoparticle immobilized trypsin

  7. Purification and Partial Characterization of Trypsin-Specific Proteinase Inhibitors from Pigeonpea Wild Relative Cajanus platycarpus L. (Fabaceae) Active against Gut Proteases of Lepidopteran Pest Helicoverpa armigera.

    Science.gov (United States)

    Swathi, Marri; Mishra, Prashant K; Lokya, Vadthya; Swaroop, Vanka; Mallikarjuna, Nalini; Dutta-Gupta, Aparna; Padmasree, Kollipara

    2016-01-01

    Proteinase inhibitors (PIs) are natural defense proteins of plants found to be active against gut proteases of various insects. A pigeonpea wild relative Cajanus platycarpus was identified as a source of resistance against Helicoverpa armigera, a most devastating pest of several crops including pigeonpea. In the light of earlier studies, trypsin-specific PIs (CpPI 63) were purified from mature dry seeds of C. platycarpus (ICPW-63) and characterized their biochemical properties in contributing to H. armigera resistance. CpPI 63 possessed significant H. armigera gut trypsin-like proteinase inhibitor (HGPI) activity than trypsin inhibitor (TI) activity. Analysis of CpPI 63 using two-dimensional (2-D) electrophoresis and matrix assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry revealed that it contained several isoinhibitors and small oligomers with masses ranging between 6 and 58 kDa. The gelatin activity staining studies suggest that these isoinhibitors and oligomers possessed strong inhibitory activity against H. armigera gut trypsin-like proteases (HGPs). The N-terminal sequence of the isoinhibitors (pI 6.6 and pI 5.6) of CpPI 63 exhibited 80% homology with several Kunitz trypsin inhibitors (KTIs) as well as miraculin-like proteins (MLPs). Further, modification of lysine residue(s) lead to 80% loss in both TI and HGPI activities of CpPI 63. In contrast, the TI and HGPI activities of CpPI 63 were stable over a wide range of temperature and pH conditions. The reported results provide a biochemical basis for pod borer resistance in C. platycarpus.

  8. Purification and Partial Characterization of Trypsin-Specific Proteinase Inhibitors from Pigeonpea Wild Relative Cajanus platycarpus L.(Fabaceae active against Gut Proteases of Lepidopteran pest Helicoverpa armigera

    Directory of Open Access Journals (Sweden)

    Marri Swathi

    2016-09-01

    Full Text Available AbstractProteinase inhibitors (PIs are natural defense proteins of plants found to be active against gut proteases of various insects. A pigeonpea wild relative Cajanus platycarpus was identified as a source of resistance against Helicoverpa armigera, a most devastating pest of several crops including pigeonpea. In the light of earlier studies, trypsin-specific PIs (CpPI 63 were purified from mature dry seeds of C. platycarpus (ICPW-63 and characterized their biochemical properties in contributing to H. armigera resistance. CpPI 63 possessed significant H. armigera gut trypsin-like proteinase inhibitor (HGPI activity than trypsin inhibitor (TI activity. Analysis of CpPI 63 using two-dimensional (2-D electrophoresis and matrix assisted laser desorption ionization time-of-flight (MALDI-TOF mass spectrometry revealed that it contained several isoinhibitors and small oligomers with masses ranging between 6-58 kDa. The gelatin activity staining studies suggest that these isoinhibitors and oligomers possessed strong inhibitory activity against H. armigera gut trypsin-like proteases (HGPs. The N-terminal sequence of the isoinhibitors (pI 6.6 and pI 5.6 of CpPI 63 exhibited 80% homology with several Kunitz trypsin inhibitors (KTIs as well as miraculin-like proteins (MLPs. Further, modification of lysine residue(s lead to 80% loss in both TI and HGPI activities of CpPI 63. In contrast, the TI and HGPI activities of CpPI 63 were stable over a wide range of temperature and pH conditions. The reported results provide a biochemical basis for pod borer resistance in C. platycarpus.

  9. Violence against women.

    Science.gov (United States)

    Shiva, M

    1999-01-01

    In India, violence against women is increasing and takes many forms while laws to protect women are ignored. Despite this fact, the new reproductive and child health program ignores sexual violence. Health personnel can respond by: 1) accepting the magnitude of the problem; 2) investigating the deaths of young women; 3) documenting findings; 4) ensuring that sexual abuse is recognized as a public health problem; 5) disseminating findings; 6) ensuring the protection of female field workers; 7) recognizing violence as an occupational health hazard; 8) facilitating the empowerment of women; 9) training women in self-defense; 10) ensuring that colleges and training institutes address violence as a women's health concern; 11) studying the psychological effects of violence; 12) collaborating with the National Commission for Women and the National Human Rights Commission; and 13) advocating for incorporation of sexual violence as a reproductive health issue in the national reproductive health program. In particular, domestic violence is a pervasive violation of women's human rights and has been resistant to social advances because of its "hidden" nature. Domestic violence exists because husbands believe they have an absolute right over the sexuality of their wives. Abusive husbands also abuse their daughters while sons learn violent behavior from their fathers. Crimes must be considered irrespective of whether they are committed outside or inside the home.

  10. Genital ulcers in women

    NARCIS (Netherlands)

    Bruisten, Sylvia M.

    2003-01-01

    Women who are in a low socioeconomic status are most vulnerable to genital ulcer disease (GUD). GUD is recognized as an important co-factor for acquisition of HIV. GUD etiology has been elucidated in the past decade, with the availability of multiplex polymerase chain reaction. Worldwide, herpes

  11. EPA Recognizes Charleston County School District for Reducing Food Waste

    Science.gov (United States)

    ATLANTA - Today, U.S. Environmental Protection Agency (EPA) recognized the Charleston County School District for the District's achievements in reducing food waste. The District cultivated one of the state's first student-driven commercial compostin

  12. Can You Recognize the Signs of Skin Cancer?

    Science.gov (United States)

    ... 166922.html Can You Recognize the Signs of Skin Cancer? First step: Get to know your own skin ... 2017 WEDNESDAY, June 28, 2017 (HealthDay News) -- With skin cancer the most common type of cancer in the ...

  13. Recognizing Advanced Heart Failure and Knowing Your Options

    Science.gov (United States)

    ... Disease Venous Thromboembolism Aortic Aneurysm More Recognizing Advanced Heart Failure and Knowing Your Options Updated:May 9,2017 ... need in the future. Treatment Options for Advanced Heart Failure Major Interventions Open-heart surgery: For patients with ...

  14. Ability of nucleus cochlear implantees to recognize music.

    Science.gov (United States)

    Fujita, S; Ito, J

    1999-07-01

    Eight adults with cochlear implants participated in experiments to test their ability to recognize music. Some subjects showed good ability to recognize songs that were sung with instrumental accompaniment but poor ability to recognize songs played on an electronic keyboard without verbal cues, indicating that they were recognizing the songs by verbal cues rather than by musical qualities such as tones and melodic intervals. This conclusion was strengthened by the finding that subjects were barely able to distinguish between songs with the same rhythm and pitch range, and they showed poor ability to discriminate musical intervals. (The closest discrimination was 4 semitones.) Subjects had good ability to distinguish among the synthesized sounds of various musical instruments played on the electronic keyboard. We speculate that subjects could distinguish the various musical instruments in the same way they distinguish among human voices using spectrographic patterns such as formants or maxima.

  15. Recognizing the Symptoms of Worsening Heart Valve Disease

    Science.gov (United States)

    ... be hard to separate our habits from the mechanics of our hearts, which is one important reason ... About Heart Valves • Heart Valve Problems and Causes • Risks, Signs and Symptoms Introduction Understanding Symptoms Recognizing Symptoms ...

  16. EPA Recognizes Bowdoin College for Energy Efficiency Efforts

    Science.gov (United States)

    Bowdoin College in Brunswick, Maine is being recognized by EPA with a 2015 Energy Star Combined Heat and Power (CHP) award. CHP technology produces electricity and useful heat simultaneously from a single energy source, such as natural gas,

  17. A novel approach in recognizing magnetic material with simplified algorithm

    KAUST Repository

    Talukdar, Abdul Hafiz Ibne

    2011-04-01

    In this article a cost-effective and simple system (circuit and algorithm) which allows recognizing different kinds of films by their magneto-field conductive properties is demonstrated. The studied signals are generated by a proposed circuit. This signal was further analyzed (recognized) in frequency domain creating the Fourier frequency spectrum which is easily used to detect the response of magnetic sample. The novel algorithm in detecting magnetic field is presented here with both simulation and experimental results. © 2011 IEEE.

  18. Discovering Activities to Recognize and Track in a Smart Environment

    OpenAIRE

    Rashidi, Parisa; Cook, Diane J.; Holder, Lawrence B.; Schmitter-Edgecombe, Maureen

    2011-01-01

    The machine learning and pervasive sensing technologies found in smart homes offer unprecedented opportunities for providing health monitoring and assistance to individuals experiencing difficulties living independently at home. In order to monitor the functional health of smart home residents, we need to design technologies that recognize and track activities that people normally perform as part of their daily routines. Although approaches do exist for recognizing activities, the approaches ...

  19. B2 kinin receptor activation is the predominant mechanism by which trypsin mediates endothelium-dependent relaxation in bovine coronary arteries.

    Science.gov (United States)

    Drummond, Grant R; Selemidis, Stavros; Cocks, Thomas M

    2008-07-01

    The roles of kinin and protease-activated receptors (PAR) in endothelium-dependent relaxations to the serine protease, trypsin, were examined in rings of bovine left anterior descending coronary artery (LAD). Trypsin (0.01-30 U/ml) caused biphasic, endothelium-dependent relaxations-a high potency (0.01-0.3 U/ml), low efficacy relaxation [maximum relaxation (R (max)), 9.0 +/- 5.1%] followed by a lower potency (1-30 U/ml) but high efficacy (R (max), 90.4 +/- 5.5%) relaxation, which was abolished by aprotinin. Captopril (10 microM) caused an approximately 10-fold leftward shift of the second phase response such that the first phase was masked. The second phase relaxation to trypsin was inhibited in a concentration-dependent, non-surmountable manner by the B2 antagonist, HOE-140. At 3 nM HOE-140, the second phase response to trypsin was abolished unmasking the first phase. Kallikrein (0.0003-0.3 U/ml) caused monophasic, endothelium-dependent relaxations (R (max), 33.7 +/- 14.6%), which were potentiated by captopril (R (max), 94.2 +/- 1.0%) and abolished by HOE-140. In the presence of captopril, the second phase relaxation to trypsin was only minimally inhibited by either N(G)-nitro-L: -arginine (100 microM) or 67 mM [K(+)](o) alone but markedly reduced when these two treatments were combined (R (max), 26.1 +/- 11.6% versus 98.6 +/- 2.9% in controls). The PAR1-activating peptide, SFLLRN (0.1-30 microM), but not the PAR2-activating peptide, SLIGRL, caused concentration-dependent relaxations (pEC(50), 5.9 +/- 0.0%; R (max), 43.3 +/- 8.3%). In conclusion, trypsin causes endothelium-dependent relaxations in the bovine LAD predominantly via release of endogenous BK, which in turn activates endothelial B2 receptors. Only a minor role for PAR1-like receptors was evident in this tissue.

  20. Elimination of trypsin inhibitor activity and beany flavor in soy milk by consecutive blanching and ultrahigh-temperature (UHT) processing.

    Science.gov (United States)

    Yuan, Shaohong; Chang, Sam K C; Liu, Zhisheng; Xu, Baojun

    2008-09-10

    Soy foods contain significant health-promoting components but also may contain beany flavor and trypsin inhibitor activity (TIA), which can cause pancreatic disease if present at a high level. Thermal processing can inactivate TIA and lipoxygenase. Ultrahigh-temperature (UHT) processing is relatively new for manufacturing soy milk. Simultaneous elimination of TIA and soy odor by UHT processing for enhancing soy milk quality has not been reported. The objective was to determine TIA in soy milk processed by traditional, steam injection, blanching, and UHT methods and to compare the products with commercial soy milk products. Soybean was soaked and blanched at 70-85 degrees C for 30 s-7.5 min. The blanched beans were made into base soy milk. The hexanal content of the base soy milk was determined by gas chromatography to determine the best conditions for further thermal processing by indirect and direct UHT methods at 135-150 degrees C for 10-50 s using the Microthermics processor. Soy milk was also made from soaked soybeans by traditional batch cooking and steaming methods. Eighteen commercial products were selected from the supermarket. Residual TIA in soy milk processed by the traditional and steam injection to 100 degrees C for 20 min was approximately 13%. Blanching could inactivate 25-50% of TIAs of the raw soy milk. The blanch conditions of 80 degrees C and 2 min were selected for UHT processing because these conditions produced blanched soy milk without hexanal, indicating a complete heat inactivation of lipoxygenases. The TIA decreased with increased temperature and time of UHT heating. The maximal trypsin inhibitor inactivation was achieved by UHT direct and indirect methods with residual activities of approximately 10%. Some commercial soy milk products contained high TIAs. The results are important to the food industry and consumers. Kinetic analysis showed that heat inactivation (denaturation) of TIA, under the continuous processing conditions of the

  1. pH titration of native and unfolded ß-trypsin: evaluation of the D D G0 titration and the carboxyl pK values

    Directory of Open Access Journals (Sweden)

    Günther A.R.

    1997-01-01

    Full Text Available The stabilizing free energy of ß-trypsin was determined by hydrogen ion titration. In the pH range from 3.0 to 7.0, the change in free energy difference for the stabilization of the native protein relative to the unfolded one (D D G0 titration was 9.51 ± 0.06 kcal/mol. An isoelectric point of 10.0 was determined, allowing us to calculate the Tanford and Kirkwood electrostatic factor w. This factor presented a nonlinear behavior and indicated more than one type of titratable carboxyl groups in ß-trypsin. In fact, one class of carboxyl group with a pK = 3.91 ± 0.01 and another one with a pK = 4.63 ± 0.03 were also found by hydrogen ion titration of the protein in the folded state

  2. Effective removal of alginate-poly-L-lysine microcapsules from pancreatic islets by use of trypsin-EDTA.

    Science.gov (United States)

    de Groot, Martijn; Leuvenink, Henri G D; Keizer, Paula P M; Fekken, Susan; Schuurs, Theo A; van Schilfgaarde, Reinout

    2003-11-01

    Although the transplantation of alginate-poly-L-lysine-alginate encapsulated islets of Langerhans usually is successful, graft survival is still limited. Molecular analysis by RT-PCR of the encapsulated islets may provide insight into the mechanisms that affect islets during graft failure. However, RT-PCR on encapsulated islets is not possible because the poly-L-lysine of the capsule interferes with both cDNA synthesis and PCR amplification. We applied a method that mechanically removes the microcapsules from the islets after a short trypsin-EDTA treatment (decapsulation), thereby enabling RT-PCR analysis. The results of this study show that the decapsulation procedure does not affect islet vitality and has only minor effects on islet function and morphology. The decapsulation does not affect GAPDH, beta-actin, Bcl-2, or Bax gene expression. This method is an improvement over the time-consuming manual dissection of microcapsules because it allows for the rapid and relatively harmless removal of capsules on a larger scale. Decapsulation offers the possibility of applying RT-PCR, as well as other methods, which cannot be performed on encapsulated islets. Copyright 2003 Wiley Periodicals, Inc.

  3. Tissue Kallikrein Inhibitors Based on the Sunflower Trypsin Inhibitor Scaffold - A Potential Therapeutic Intervention for Skin Diseases.

    Science.gov (United States)

    Chen, Wenjie; Kinsler, Veronica A; Macmillan, Derek; Di, Wei-Li

    2016-01-01

    Tissue kallikreins (KLKs), in particular KLK5, 7 and 14 are the major serine proteases in the skin responsible for skin shedding and activation of inflammatory cell signaling. In the normal skin, their activities are controlled by an endogenous protein protease inhibitor encoded by the SPINK5 gene. Loss-of-function mutations in SPINK5 leads to enhanced skin kallikrein activities and cause the skin disease Netherton Syndrome (NS). We have been developing inhibitors based on the Sunflower Trypsin Inhibitor 1 (SFTI-1) scaffold, a 14 amino acids head-to-tail bicyclic peptide with a disulfide bond. To optimize a previously reported SFTI-1 analogue (I10H), we made five analogues with additional substitutions, two of which showed improved inhibition. We then combined those substitutions and discovered a variant (Analogue 6) that displayed dual inhibition of KLK5 (tryptic) and KLK7 (chymotryptic). Analogue 6 attained a tenfold increase in KLK5 inhibition potency with an Isothermal Titration Calorimetry (ITC) Kd of 20nM. Furthermore, it selectively inhibits KLK5 and KLK14 over seven other serine proteases. Its biological function was ascertained by full suppression of KLK5-induced Protease-Activated Receptor 2 (PAR-2) dependent intracellular calcium mobilization and postponement of Interleukin-8 (IL-8) secretion in cell model. Moreover, Analogue 6 permeates through the cornified layer of in vitro organotypic skin equivalent culture and inhibits protease activities therein, providing a potential drug lead for the treatment of NS.

  4. Silica nanoparticle-based microfluidic immunosensor with laser-induced fluorescence detection for the quantification of immunoreactive trypsin.

    Science.gov (United States)

    Seia, Marco A; Stege, Patricia W; Pereira, Sirley V; De Vito, Irma E; Raba, Julio; Messina, Germán A

    2014-10-15

    The purpose of this study was to develop a silica nanoparticle-based immunosensor with laser-induced fluorescence (LIF) as a detection system. The proposed device was applied to quantify the immunoreactive trypsin (IRT) in cystic fibrosis (CF) newborn screening. A new ultrasonic procedure was used to extract the IRT from blood spot samples collected on filter papers. After extraction, the IRT reacted immunologically with anti-IRT monoclonal antibodies immobilized on a microfluidic glass chip modified with 3-aminopropyl functionalized silica nanoparticles (APSN-APTES-modified glass chips). The bounded IRT was quantified by horseradish peroxidase (HRP)-conjugated anti-IRT antibody (anti-IRT-Ab) using 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) as enzymatic mediator. The HRP catalyzed the oxidation of nonfluorescent ADHP to highly fluorescent resorufin, which was measured by LIF detector, using excitation lambda at 561nm and emission at 585nm. The detection limits (LODs) calculated for LIF detection and for a commercial enzyme-linked immunosorbent assay (ELISA) test kit were 0.87 and 4.2ngml(-1), respectively. The within- and between-assay variation coefficients for the LIF detection procedure were below 6.5%. The blood spot samples collected on filter papers were analyzed with the proposed method, and the results were compared with those of the reference ELISA method, demonstrating a potential usefulness for the clinical assessment of IRT during the early neonatal period. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. Pressurized Pepsin Digestion in Proteomics: An Automatable Alternative to Trypsin for Integrated Top-down Bottom-up Proteomics

    Energy Technology Data Exchange (ETDEWEB)

    Lopez-Ferrer, Daniel; Petritis, Konstantinos; Robinson, Errol W.; Hixson, Kim K.; Tian, Zhixin; Lee, Jung Hwa; Lee, Sang-Won; Tolic, Nikola; Weitz, Karl K.; Belov, Mikhail E.; Smith, Richard D.; Pasa-Tolic, Ljiljana

    2011-02-01

    Integrated top-down bottom-up proteomics combined with online digestion has great potential to improve the characterization of protein isoforms in biological systems and is amendable to highthroughput proteomics experiments. Bottom-up proteomics ultimately provides the peptide sequences derived from the tandem MS analyses of peptides after the proteome has been digested. Top-down proteomics conversely entails the MS analyses of intact proteins for more effective characterization of genetic variations and/or post-translational modifications (PTMs). Herein, we describe recent efforts towards efficient integration of bottom-up and top-down LCMS based proteomic strategies. Since most proteomic platforms (i.e. LC systems) operate in acidic environments, we exploited the compatibility of the pepsin (i.e. the enzyme’s natural acidic activity) for the integration of bottom-up and top-down proteomics. Pressure enhanced pepsin digestions were successfully performed and characterized with several standard proteins in either an offline mode using a Barocycler or an online mode using a modified high pressure LC system referred to as a fast online digestion system (FOLDS). FOLDS was tested using pepsin and a whole microbial proteome, and the results compared against traditional trypsin digestions on the same platform. Additionally, FOLDS was integrated with a RePlay configuration to demonstrate an ultra-rapid integrated bottom-up top-down proteomic strategy employing a standard mixture of proteins and a monkey pox virus proteome.

  6. Effects of Polymorphisms in Pepsinogen (PEP, Amylase (AMY and Trypsin (TRY Genes on Food Habit Domestication Traits in Mandarin Fish

    Directory of Open Access Journals (Sweden)

    Tilin Yi

    2013-10-01

    Full Text Available Mandarin fish (Siniperca chuatsi have a peculiar feeding habit of only accepting live fish prey and refusing dead prey and artificial diets. However, previous research has shown that some individuals accept dead prey after gradual domestication. Digestive enzymes are correlated with feeding habits in fish. In the current study, SNPs in the mandarin fish genes for pepsinogen (PEP, amylase (AMY, and trypsin (TRY were evaluated for associations with feeding habits in domesticated mandarin fish by scanning their complete genomic sequence. In total, two SNPs were found in PEP, one was found in TRY, and none were found in AMY. The D1(CTCC and D5(TTTT diplotypes in the PEP gene tended to show strong effects on the feeding habits of domesticated fish (p < 0.01. The results indicate that PEP may be associated with the genetic mechanism for feeding habits in mandarin fish, and the D1(CTCC and D5(TTTT diplotypes in the PEP gene may be useful markers for selecting mandarin fish with appropriate feeding habits for domestication.

  7. Selective Isolation of Trypsin Inhibitor and Lectin from Soybean Whey by Chitosan/Tripolyphosphate/Genipin Co-Crosslinked Beads

    Directory of Open Access Journals (Sweden)

    Yu-Lung Chang

    2014-06-01

    Full Text Available Selective isolation of Kunitz trypsin inhibitor (KTI and lectin from soybean whey solutions by different types of chitosan beads was investigated. The chitosan beads were co-crosslinked with tripolyphosphate/genipin in solutions at pH 5, 7 or 9 (CB5, CB7, CB9. The maximum adsorption ratios of chitosan beads to KTI and lectin were observed at pH 4.4 and 5.4, respectively; highly selective separation was also demonstrated at these pHs. The adsorption ratios increased with temperature, rising between 5 and 25 °C. CB9 produced the best adsorption ratio, followed by CB7 then CB5. The critical interaction governing absorption of chitosan beads to KTI and lectin could be hydrogen bonding. At pH 9, KTI and lectin desorbed efficiently from CB7 with desorption ratios of 80.9% and 81.4%, respectively. The desorption was most likely caused predominantly by electrostatic repulsion. KTI and lectin can effectively be selectively isolated from soybean whey using this novel separation technique.

  8. A lectin affinity workflow targeting glycosite-specific, cancer-related carbohydrate structures in trypsin-digested human plasma.

    Science.gov (United States)

    Drake, Penelope M; Schilling, Birgit; Niles, Richard K; Braten, Miles; Johansen, Eric; Liu, Haichuan; Lerch, Michael; Sorensen, Dylan J; Li, Bensheng; Allen, Simon; Hall, Steven C; Witkowska, H Ewa; Regnier, Fred E; Gibson, Bradford W; Fisher, Susan J

    2011-01-01

    Glycans are cell-type-specific, posttranslational protein modifications that are modulated during developmental and disease processes. As such, glycoproteins are attractive biomarker candidates. Here, we describe a mass spectrometry-based workflow that incorporates lectin affinity chromatography to enrich for proteins that carry specific glycan structures. As increases in sialylation and fucosylation are prominent among cancer-associated modifications, we focused on Sambucus nigra agglutinin (SNA) and Aleuria aurantia lectin (AAL), lectins which bind sialic acid- and fucose-containing structures, respectively. Fucosylated and sialylated glycopeptides from human lactoferrin served as positive controls, and high-mannose structures from yeast invertase served as negative controls. The standards were spiked into Multiple Affinity Removal System (MARS) 14-depleted, trypsin-digested human plasma from healthy donors. Samples were loaded onto lectin columns, separated by HPLC into flow-through and bound fractions, and treated with peptide: N-glycosidase F to remove N-linked glycans. The deglycosylated peptide fractions were interrogated by ESI HPLC-MS/MS. We identified a total of 122 human plasma glycoproteins containing 247 unique glycosites. Importantly, several of the observed glycoproteins (e.g., cadherin 5 and neutrophil gelatinase-associated lipocalin) typically circulate in plasma at low nanogram per milliliter levels. Together, these results provide mass spectrometry-based evidence of the utility of incorporating lectin-separation platforms into cancer biomarker discovery pipelines. Copyright © 2010 Elsevier Inc. All rights reserved.

  9. Gender Discrimination and Women's Development in India

    Science.gov (United States)

    Sivakumar, M.

    2008-01-01

    Gender is a common term where as gender discrimination is meant only for women, because females are the only victims of gender discrimination. Females are nearly 50 percent of the total population but their representation in public life is very low. Recognizing women's right and believing their ability are essential for women's empowerment and…

  10. Anxiety and ability to recognize clinical information in dentistry.

    Science.gov (United States)

    Eli, I; Schwartz-Arad, D; Bartal, Y

    2008-01-01

    Stress significantly affects a person's cognitive ability to process information. Therefore, we hypothesized that patients' ability to recognize information related to the procedure they are about to undergo will be affected by the stressfulness of the situation (less recognition under a high-stress situation as compared with a low-stress situation). Patients (n = 66) were evaluated for their ability to recognize clinical information supplied on two different occasions: immediately before oral surgery (high-stress condition) and before suture removal (low-stress condition). Dental and state of anxiety and expectation of pain were also assessed. On both occasions, the patients' ability to recognize information correctly was low (less than 50%). Patients recognized significantly less information pre-operatively than before suture removal. State of anxiety, dental anxiety, and expectation to experience pain had a profound effect on their ability to recognize provided information correctly. Apparently, before dental treatment (high or low on stress), patients' ability to process information may be severely impaired.

  11. Effect of low-dose irradiation on soybean protein solubility, trypsin inhibitor activity, and protein patterns separated by polyacrylamide gel electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Afify, M.R.; Shousha, Moustafa A. (Cairo Univ., Giza (Egypt))

    Five soybean cultivars were used in this study to investigate changes in soybean protein solubility, protein patterns separated by SDE-PAGE and Poro-PAGE, and activity of trypsin inhibitor after exposure to {gamma} irradiation. SDS-PAGE proved that changes in protein subunits patterns could by identified in the Clark cultivar. Three high molecular weight protein bands were detected in irradiated soybean cultivars by using Poro-PAGE.

  12. Purification and Partial Characterization of Trypsin-Specific Proteinase Inhibitors from Pigeonpea Wild Relative Cajanus platycarpus L. (Fabaceae) Active against Gut Proteases of Lepidopteran Pest Helicoverpa armigera

    OpenAIRE

    Marri Swathi; Mishra, Prashant K.; Vadthya Lokya; Swaroop Vanka; Nalini Mallikarjuna; Aparna Dutta Gupta; Kollipara Padmasree

    2016-01-01

    AbstractProteinase inhibitors (PIs) are natural defense proteins of plants found to be active against gut proteases of various insects. A pigeonpea wild relative Cajanus platycarpus was identified as a source of resistance against Helicoverpa armigera, a most devastating pest of several crops including pigeonpea. In the light of earlier studies, trypsin-specific PIs (CpPI 63) were purified from mature dry seeds of C. platycarpus (ICPW-63) and characterized their biochemical properties in cont...

  13. Follicle cell trypsin-like protease HrOvochymase: Its cDNA cloning, localization, and involvement in the late stage of oogenesis in the ascidian Halocynthia roretzi.

    Science.gov (United States)

    Mino, Masako; Sawada, Hitoshi

    2016-04-01

    We previously reported that the sperm trypsin-like protease HrAcrosin and its precursor HrProacrosin participate in fertilization of the ascidian Halocynthia roretzi. The HrProacrosin gene is annotated in the H. roretzi genome database as Harore.CG.MTP2014.S89.g15383; our previously reported sequence of HrProacrosin gene appeared to include four nucleotides inserted near the 3'-end of HrProacrosin, resulting in a frame-shift mutation and a premature termination codon. The gene architecture of HrProacrosin and Harore.CG.MTP2014.S89.g15383 resembles that of Xenopus laevis ovochymase-1/OVCH1 and ovochymase-2/OVCH2, which encode egg extracellular polyproteases. Considering these new observations, we evaluated the cDNA cloning, expression, localization, and function of Harore.CG.MTP2014.S89.g15383, herein designated as HrOvochymase/HrOVCH. We found that HrOVCH cDNA consists of a single open reading frame of 1,575 amino acids, containing a signal peptide, three trypsin-like protease domains, and six CUB domains. HrOVCH was transcribed by the testis and ovary, but the majority of protein exists in ovarian follicle cells surrounding eggs. An anti-HrOVCH antibody inhibited elevation of the vitelline coat at a late stage of oogenesis, during the period when self-sterility is acquired. As trypsin inhibitors are reported to block the acquisition of self-sterility during oogenesis, whereas trypsin induces the acquisition of self-sterility and elevation of the vitelline coat in defolliculated ovarian eggs, we propose that HrOVCH may play a role in the acquisition of self-sterility by late-stage H. roretzi oocytes. © 2016 Wiley Periodicals, Inc.

  14. Supplementation with a new trypsin inhibitor from peanut is associated with reduced fasting glucose, weight control, and increased plasma CCK secretion in an animal model.

    Science.gov (United States)

    Serquiz, Alexandre C; Machado, Richele J A; Serquiz, Raphael P; Lima, Vanessa C O; de Carvalho, Fabiana Maria C; Carneiro, Marcella A A; Maciel, Bruna L L; Uchôa, Adriana F; Santos, Elizeu A; Morais, Ana H A

    2016-12-01

    Ingestion of peanuts may have a beneficial effect on weight control, possibly due to the satietogenic action of trypsin inhibitors. The aim of this study was to isolate a new trypsin inhibitor in a typical Brazilian peanut sweet (paçoca) and evaluate its effect in biochemical parameters, weight gain and food intake in male Wistar rats. The trypsin inhibitor in peanut paçoca (AHTI) was isolated. Experimental diets were prepared with AIN-93G supplemented with AHTI. Animals had their weight and food intake monitored. Animals were anesthetized, euthanized, and their bloods collected by cardiac puncture for dosage of cholecystokinin (CCK) and other biochemical parameters. Supplementation with AHTI significantly decreased fasting glucose, body weight gain, and food intake. These effects may be attributed to increased satiety, once supplemented animals showed no evidence of impaired nutritional status and also because AHTI increased CCK production. Thus, our results indicate that AHTI, besides reducing fasting glucose, can reduce weight gain via food intake reduction.

  15. A case of neofunctionalization of a Putranjiva roxburghii PNP protein to trypsin inhibitor by disruption of PNP-UDP domain through an insert containing inhibitory site.

    Science.gov (United States)

    Verma, Preeti; Patel, Girijesh K; Kar, Bibekananda; Sharma, Ashwani K

    2017-07-01

    The attainment of new function by a protein is achieved through convergent/divergent evolution. In present work, the sequence analysis of a 34kDa protein from Putranjiva roxburghii, earlier reported as a potent trypsin inhibitor, showed resemblance to some of the wound inducible and vegetative storage proteins. A detailed sequence analysis revealed that these proteins belong to PNP-UDP family. In case of P. roxburghii protein, an approximately 46 residue insert disrupts the PNP domain. Similar disruption of PNP domain is observed in related plant proteins. The characterization of recombinant full length and truncated (without 46 residue insert) forms of P. roxburghii PNP family protein (PRpnp) unraveled that trypsin inhibitory active site is located within the insert. The truncated form containing uninterrupted PNP domain showed strong PNP enzymatic activity where it hydrolyzed the N-glycosidic bond of inosine and guanosine. The full length protein, however, showed weak PNP enzyme activity which may be due to presence of the insert. These results indicate towards the neofunctionalization of PRpnp to a potent trypsin inhibitor through an insert containing inhibitory residue to cater to the needs of plant defense. The similar wound inducible and vegetative storage proteins may have also evolved due to evolutionary needs. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. A Trypsin Inhibitor from Tamarind Reduces Food Intake and Improves Inflammatory Status in Rats with Metabolic Syndrome Regardless of Weight Loss

    Directory of Open Access Journals (Sweden)

    Fabiana M. C. Carvalho

    2016-09-01

    Full Text Available Trypsin inhibitors are studied in a variety of models for their anti-obesity and anti-inflammatory bioactive properties. Our group has previously demonstrated the satietogenic effect of tamarind seed trypsin inhibitors (TTI in eutrophic mouse models and anti-inflammatory effects of other trypsin inhibitors. In this study, we evaluated TTI effect upon satiety, biochemical and inflammatory parameters in an experimental model of metabolic syndrome (MetS. Three groups of n = 5 male Wistar rats with obesity-based MetS received for 10 days one of the following: (1 Cafeteria diet; (2 Cafeteria diet + TTI (25 mg/kg; and (3 Standard diet. TTI reduced food intake in animals with MetS. Nevertheless, weight gain was not different between studied groups. Dyslipidemia parameters were not different with the use of TTI, only the group receiving standard diet showed lower very low density lipoprotein (VLDL and triglycerides (TG (Kruskal–Wallis, p < 0.05. Interleukin-6 (IL-6 production did not differ between groups. Interestingly, tumor necrosis factor-alpha (TNF-α was lower in animals receiving TTI. Our results corroborate the satietogenic effect of TTI in a MetS model. Furthermore, we showed that TTI added to a cafeteria diet may decrease inflammation regardless of weight loss. This puts TTI as a candidate for studies to test its effectiveness as an adjuvant in MetS treatment.

  17. Immobilization of trypsin in the layer-by-layer coating of graphene oxide and chitosan on in-channel glass fiber for microfluidic proteolysis.

    Science.gov (United States)

    Bao, Huimin; Chen, Qiwen; Zhang, Luyan; Chen, Gang

    2011-12-21

    In this report, trypsin was immobilized in the layer-by-layer (LBL) coating of graphene oxide (GO) and chitosan on a piece of glass fiber to fabricate microchip bioreactor for efficient proteolysis. LBL deposition driven by electrostatic forces easily took place on the surface of the glass fiber, providing mild environmental conditions so that the denaturation and autolysis of the immobilized trypsin was minimized. Prior to use, a piece of the prepared trypsin-immobilized glass fiber was inserted into the channel of a microchip to form a core-changeable bioreactor. The novel GO-based bioreactor can be regenerated by changing its fiber core. The feasibility and performance of the unique bioreactor were demonstrated by the tryptic digestion of bovine serum albumin, myoglobin, cytochrome c, and hemoglobin and the digestion time was significantly reduced to less than 10 s. The obtained digests were identified by MALDI-TOF MS. The digestion performance of the core-changeable GO-based microchip bioreactor was comparable to that of 12-h in-solution tryptic digestion. The novel microchip bioreactor is simple and efficient, offering great promise for high-throughput protein identification.

  18. Recognizing online Arabic handwritten characters using a deep architecture

    Science.gov (United States)

    Tagougui, Najiba; Kherallah, Monji

    2017-03-01

    Recognizing the online Arabic handwritten script has been gaining more interest because of the impressive advances in mobile device requiring more and more intelligent handwritten recognizers. Since it was demonstrated within many previous research that Deep Neural Networks (DNN) exhibit a great performance, we propose in this work a new system based on a DNN in which we try to optimize the training process by a smooth construct of the deep architecture. The Output's error of each unit in the previous layer will be computed and only the smallest error will be maintained in the next iteration. This paper uses LMCA database for training and testing data. The experimental study reveals that our proposed DBNN using generated Bottleneck features can outperform state of the art online recognizers.

  19. Recognizing Variable Environments The Theory of Cognitive Prism

    CERN Document Server

    Dong, Tiansi

    2012-01-01

    Normal adults do not have any difficulty in recognizing their homes. But can artificial systems do in the same way as humans? This book collects interdisciplinary evidences and presents an answer from the perspective of computing, namely, the theory of cognitive prism. To recognize an environment, an intelligent system only needs to classify objects, structures them based on the connection relation (not through measuring!), subjectively orders the objects, and compares with the target environment, whose knowledge is similarly structured. The intelligent system works, therefore, like a prism: when a beam of light (a scene) reaches (is perceived) to an optical prism (by an intelligent system), some light (objects) is reflected (are neglected), those passed through (the recognized objects) are distorted (are ordered differently). So comes the term 'cognitive prism'! Two fundamental propositions used in the theory can be informally stated as follow: an orientation relation is a kind of distance comparison relatio...

  20. You look familiar: how Malaysian Chinese recognize faces.

    Science.gov (United States)

    Tan, Chrystalle B Y; Stephen, Ian D; Whitehead, Ross; Sheppard, Elizabeth

    2012-01-01

    East Asian and white Western observers employ different eye movement strategies for a variety of visual processing tasks, including face processing. Recent eye tracking studies on face recognition found that East Asians tend to integrate information holistically by focusing on the nose while white Westerners perceive faces featurally by moving between the eyes and mouth. The current study examines the eye movement strategy that Malaysian Chinese participants employ when recognizing East Asian, white Western, and African faces. Rather than adopting the Eastern or Western fixation pattern, Malaysian Chinese participants use a mixed strategy by focusing on the eyes and nose more than the mouth. The combination of Eastern and Western strategies proved advantageous in participants' ability to recognize East Asian and white Western faces, suggesting that individuals learn to use fixation patterns that are optimized for recognizing the faces with which they are more familiar.

  1. You look familiar: how Malaysian Chinese recognize faces.

    Directory of Open Access Journals (Sweden)

    Chrystalle B Y Tan

    Full Text Available East Asian and white Western observers employ different eye movement strategies for a variety of visual processing tasks, including face processing. Recent eye tracking studies on face recognition found that East Asians tend to integrate information holistically by focusing on the nose while white Westerners perceive faces featurally by moving between the eyes and mouth. The current study examines the eye movement strategy that Malaysian Chinese participants employ when recognizing East Asian, white Western, and African faces. Rather than adopting the Eastern or Western fixation pattern, Malaysian Chinese participants use a mixed strategy by focusing on the eyes and nose more than the mouth. The combination of Eastern and Western strategies proved advantageous in participants' ability to recognize East Asian and white Western faces, suggesting that individuals learn to use fixation patterns that are optimized for recognizing the faces with which they are more familiar.

  2. Women's rights are human rights -- why development has failed women.

    Science.gov (United States)

    1995-08-01

    Oxfam UK/I believes that all women have the right to a livelihood, reproductive choice, health care, education, and employment. Access to resources, decision-making, political and religious freedom of expression, and freedom from all forms of violence are also equally important. Oxfam UK/I recognizes that women worldwide from a diversity of cultures and religions are arguing for similar rights, but continuing efforts to create women's equality and empowerment have had only limited success. There has been no significant improvement in women's lifestyles, the feminization of poverty is increasing, growing religious fundamentalism threatens advances made toward equality, and there has been an increasing violation of women's individual human rights to development in the last decade. Human rights instruments to tackle gender inequality exist, but they are not implemented. The rights approach to development recognizes that unless inequalities at local, national, and international levels are seen and challenged, women will continue to face poverty, inadequate representation of their needs and views, and policies which are contrary to their interests. The rights approach encompasses all aspects of women's lives, recognizing that women's rights in the civil, political, social, economic, and cultural spheres are indivisible from one another in the realities of daily life. Oxfam UK/I acknowledges the complexity of the rights debate.

  3. CDC Vital Signs: Binge Drinking a Serious, Under-Recognized Problem Among Women and Girls

    Science.gov (United States)

    ... US Dietary Guidelines on alcohol consumption (see box). Helping states and communities to report on how many people binge drink, how often and how much they drink when they binge, and whether health care providers are screening and counseling for excessive alcohol use. Reporting on youth exposure ...

  4. Recognizing Family Dynamics in the Treatment of Chronic Fatigue Syndrome

    Science.gov (United States)

    Sperry, Len

    2012-01-01

    Chronic fatigue syndrome (CFS) is an increasingly common chronic medical condition that affects not only patients but also their families. Because family dynamics, particularly the family life cycle, can and does influence the disease process, those providing counseling to CFS patients and their families would do well to recognize these dynamics.…

  5. Recognizing Risk-of-Failure in Communication Design Projects

    Science.gov (United States)

    Yee, Joyce; Lievesley, Matthew; Taylor, Louise

    2009-01-01

    The pace of commercial graphic design practice presents very few opportunities to conduct user research after a project's launch. This makes the design team's ability to anticipate and address risks during the design development phase even more important, recognized in the astute observation from Tim Brown, CEO of leading international design…

  6. Recognizing and Fostering Creativity in Technological Design Education

    Science.gov (United States)

    Cropley, David; Cropley, Arthur

    2010-01-01

    The importance of creativity in technological design education is now clearly recognized, both in everyday understanding and also in formal curriculum guidelines. Design offers special opportunities for creativity because of the "openness" of problems (ill-defined problems, the existence of a variety of pathways to the solution, the absence of…

  7. Engaging Others in Recognizing the Benefits of Physical Activity

    Science.gov (United States)

    Wilson-Graham, Dianne

    2007-01-01

    Recent research that examines the relationship between physical activity and academic performance provides physical educators with multiple opportunities to engage others in recognizing the benefits of physical activity and high quality physical education programs. Local schools and community provide the greatest opportunity to educate and…

  8. Computer Program Recognizes Patterns in Time-Series Data

    Science.gov (United States)

    Hand, Charles

    2003-01-01

    A computer program recognizes selected patterns in time-series data like digitized samples of seismic or electrophysiological signals. The program implements an artificial neural network (ANN) and a set of N clocks for the purpose of determining whether N or more instances of a certain waveform, W, occur within a given time interval, T. The ANN must be trained to recognize W in the incoming stream of data. The first time the ANN recognizes W, it sets clock 1 to count down from T to zero; the second time it recognizes W, it sets clock 2 to count down from T to zero, and so forth through the Nth instance. On the N + 1st instance, the cycle is repeated, starting with clock 1. If any clock has not reached zero when it is reset, then N instances of W have been detected within time T, and the program so indicates. The program can readily be encoded in a field-programmable gate array or an application-specific integrated circuit that could be used, for example, to detect electroencephalographic or electrocardiographic waveforms indicative of epileptic seizures or heart attacks, respectively.

  9. Subsistence fishers were legally recognized in South Africa for the ...

    African Journals Online (AJOL)

    denise

    Subsistence fishers were legally recognized in South. Africa for the first time in 1998 with the promulgation of the Marine Living Resources Act (MLRA). Prior to this Act, fishers harvesting marine resources for purposes other than recreational or commercial fishing were regulated through law enforcement and were.

  10. 40 CFR 105.15 - How are award winners recognized?

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 21 2010-07-01 2010-07-01 false How are award winners recognized? 105.15 Section 105.15 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER... ceremony as recognition for an outstanding technological achievement or an innovative process, method or...

  11. Development Of Software To Recognize Parts Of Plants

    Science.gov (United States)

    Despain, Ronald R.; Tharpe, Roy, Jr.; Davis, Leon; Hauss, Sharon; Shawaga, Larry; Biro, Ron

    1993-01-01

    Report describes first phase in development of digital image-processing subsystem recognizing parts of plants. Subsystem part of robotic system tending and harvesting plants in automated plant-growth chamber. Initial focus on image-processing software that distinguishes among seed heads, stems, and leaves of wheat plants and further distinguishes between these parts and background. Software adaptable to other types of plants.

  12. Self glycosphingolipids: new antigens recognized by autoreactive T lymphocytes.

    Science.gov (United States)

    De Libero, Gennaro; Mori, Lucia

    2003-04-01

    T cells may recognize glycolipids and lipids of bacterial and self origin associated with the CD1 antigen-presenting molecules. Understanding the mechanisms governing CD1-self glycolipid interaction will provide information on the molecular rules of glycolipid presentation and suggest new approaches to immunotherapy.

  13. Professional Development Recognizing Technology Integration Modeled after the TPACK Framework

    Science.gov (United States)

    McCusker, Laura

    2017-01-01

    Public school teachers within a Pennsylvania intermediate unit are receiving inadequate job-embedded professional development that recognizes knowledge of content, pedagogy, and technology integration, as outlined by Mishra and Koehler's Technological Pedagogical Content Knowledge (TPACK) framework (2006). A school environment where teachers are…

  14. Recognizing, explaining and countering norm transgressive behaviour on social media

    NARCIS (Netherlands)

    Padje, E.D.H.

    2016-01-01

    In this thesis, it is researched how norm transgressive behaviour exhibited on the Dutch domains of social media can be recognized, explained and countered. An analysis of four comment threads is conducted, of which the comments can be found on the Facebook pages of three Dutch news sites and on a

  15. Recognizing and Developing Adaptive Expertise within Outdoor and Expedition Leaders

    Science.gov (United States)

    Tozer, Mark; Fazey, Ioan; Fazey, John

    2007-01-01

    Adaptive expertise, an individual's ability to perform flexibly and innovatively in novel and unstructured situations, could have particular relevance for expedition and outdoor leaders. This element may be recognized in leadership practitioners who are able to act more effectively when problem-solving in complex, ambiguous and unpredictable…

  16. Structural elements recognized by abacavir-induced T cells

    DEFF Research Database (Denmark)

    Yerly, Daniel; Pompeu, Yuri Andreiw; Schutte, Ryan J.

    2017-01-01

    of autoimmune destruction. The structural elements recognized by drug-specific T cell receptors (TCRs) in vivo are poorly defined. Drug-stimulated T cells express TCRs specific for peptide/HLA complexes, but the characteristics of peptides (sequence, or endogenous or exogenous origin) presented in the context...

  17. Introgression of null allele of Kunitz trypsin inhibitor through marker-assisted backcross breeding in soybean (Glycine max L. Merr.).

    Science.gov (United States)

    Maranna, Shivakumar; Verma, Khushbu; Talukdar, Akshay; Lal, Sanjay Kumar; Kumar, Anil; Mukherjee, Keya

    2016-07-12

    Presence of Kunitz trypsin inhibitor (KTI) in soybean seeds necessitates pre-heat treatment of the soy-flour for its inactivation before using it in food and feed products. The heat treatment not only enhances processing costs of the soy-based foods and feeds but also affects seed-protein quality and solubility. Genetic elimination of KTI is an important and effective approach. Therefore, molecular marker-assisted backcross breeding (MABB) approach was adopted for genetic elimination of KTI from two popular soybean genotypes, DS9712 and DS9814. PI542044, an exotic germplasm line was used as donor of the kti allele which inhibits functional KTI peptide production. Foreground selection for the kti allele was performed with three closely linked SSR markers while background selection was done with 93 polymorphic SSR markers. Plants in the BC1F1 generation were found to recover 70.4-87.63 % and 60.26-73.78 % of the recurrent parent genome (RPG) of DS9712 and DS9814, respectively. Similarly, selected plants in the BC2F1 generation had 93.01-98.92 % and 83.3-91.67 % recovery of their respective RPGs. Recombinant selection was performed so as to identify plants with minimal linkage drag. Biochemical analysis of the seeds of the selected plants (ktikti) confirmed absence of KTI peptides in the seeds. Phenotypically, the selected plants were comparable to the respective recurrent parent in yield and other traits. MABB approach helped in speedy development of 6 KTI free breeding lines of soybean. Such lines will be suitable for the farmers and the soybean industries to use in production of soy-based foods and feeds without pre-heat treatment of the soy-flour. It would contribute towards wider acceptability of soy-based foods and feeds.

  18. Safety of postoperative administration of human urinary trypsin inhibitor in lung cancer patients with idiopathic pulmonary fibrosis.

    Directory of Open Access Journals (Sweden)

    Yoshikane Yamauchi

    Full Text Available Patients with idiopathic pulmonary fibrosis (IPF undergoing pulmonary resection for lung cancer carry risks of acute exacerbations of IPF (AE postoperatively. Currently, agents which may attenuate AE are actively sought. Urinary trypsin inhibitor, ulinastatin, is a synthetic glycoprotein which may potentially inhibit various inflammatory factors associated with the development and progression of IPF. The present study was done to evaluate the effects of administration of high dose ulinastatin in lung cancer patients with IPF immediately following lung resection.Patients with IPFs radiologically diagnosed on high resolution CT, and histologically diagnosed resectable lung cancers, were eligible for the study. The effects of escalating doses of ulinastatin 3×10(5, 6×10(5, and 9×10(5 units/body/day, administered postoperatively for 3 days were evaluated. The endpoints were safety and feasibility.Nine patients were evaluated, in cohorts of 3 patients per dosage. Postoperative follow up ranged from 3 to 12 months (median 9 months. The postoperative courses were uneventful in all patients. No subjective adverse events such as abdominal symptoms or skin rashes, or objective adverse events as per serum laboratory tests, such as liver or kidney dysfunctions potentially attributable to ulinastatin administration were observed. AE was seen in one patient at 3 months after surgery, but since this occurred shortly after administration of chemotherapy, it was considered to be attributable to the chemotherapy rather than surgery.Ulinastatin administration after lung resection in lung cancer patients with IPF was considered to be safe and feasible. Further study is planned at the highest dose of this study to evaluate efficacy.UMIN.ac.jp/ctr/UMIN000002410.

  19. [Study on food safety of genetically modified rice which expressed cowpea trypsin inhibitor by 90 day feeding test on rats].

    Science.gov (United States)

    Zhuo, Qin; Chen, Xiaoping; Piao, Jianhua; Gu, Lüzhen

    2004-03-01

    Rats were fed by transgenic rice which expressed insecticidal protein CPTI (cowpea trypsin inhibitor) to study if the transgenic rice possessed potential toxic or adverse effects. Weanling Wistar rats were randomly divided into three groups: T, N and C group. The diet of T group contained 78.3% transgenic rice. The diet of N group contained 74.7% non transgenic rice which was the parent line of the transgenic one. The diet formula of C group was AIN93G. The macro- and micronutrient content were equal in three diets. The rats were fed for 90 days. Food intakes were weight every day, body-weight were weight and body-length were measured every week. In the middle and at the end of feeding period, haematological value and clinical chemistry parameters were measured, at the end of the 90th day, post-mortem organ coefficient were measured, organ tissues analysis was performed and bone density was measured. In most situation, there were no significant differences among the three groups(P > 0.05) and no histopathological damage were detected. At the end of the 1st month, the male rats' body length of the T group was longer than the other two groups and at the end of the test period, the male rats' blood glucose and ALT were lower than the other two groups. In the middle of the test period, the female rats' red blood cell number and hemoglobin were higher than the other two groups and at the end of the test period, the female rats' monocyte number was higher than the other two groups (P transgenic rice on rats there did not reveal any signs of toxic and adverse effects.

  20. Bile enhances glucose uptake, reduces permeability, and modulates effects of lectins, trypsin inhibitors and saponins on intestinal tissue.

    Science.gov (United States)

    Bakke, Anne Marie; Chikwati, Elvis M; Venold, Fredrik F; Sahlmann, Christian; Holm, Halvor; Penn, Michael H; Oropeza-Moe, Marianne; Krogdahl, Åshild

    2014-02-01

    Antinutritional factors (ANFs) can disrupt digestive and other intestinal functions. ANFs in soybean meal (SBM) are implicated in proliferative and inflammatory responses in the intestine of various (functionally) monogastric animals, including Atlantic salmon (Salmo salar L.). The goal of the current study was to investigate the effect of ex vivo exposure of mid and distal intestinal tissue of salmon to soybean saponins (SAP), lectin (LEC) and Kunitz' trypsin inhibitor (KTI), singly and in combination, on epithelial function, as assessed by measuring in vitro glucose uptake pathways along a glucose concentration gradient. As solubilization of SAP in the calcium-containing Ringer's solution was problematic but resolved with the addition of a physiological concentration of bile collected from the gall bladder of salmon, an evaluation of bile effects became an added element. Results indicated that bile increased baseline glucose absorption and possibly transport, and also had a protective effect on the epithelial barrier, at least partially due to taurocholate. Compared to controls, tissues exposed to LEC+bile, KTI+bile and LEC+KTI+bile exhibited increased glucose uptake at the higher glucose concentrations, apparently due to markedly increased tissue permeability. Addition of SAP, however, attenuated the response, possibly by binding bile components. SAP+bile, also in combination with LEC and/or KTI, as well as LEC, KTI and LEC+KTI without bile often reduced transcellular glucose uptake pathways, while maintaining low tissue permeability. SAP+LEC+KTI+bile, LEC and KTI caused the most marked reductions. The distal intestine was more affected, reflecting the restriction of in vivo SBM-induced inflammatory changes to this region. Copyright © 2013 Elsevier Inc. All rights reserved.

  1. Recognizing and treating pelvic pain and pelvic floor dysfunction.

    Science.gov (United States)

    Prather, Heidi; Spitznagle, Theresa Monaco; Dugan, Sheila A

    2007-08-01

    The reported prevalence rates of pain within the pelvis range from 3.8% to 24% in women aged 15 to 73 years. Despite the significant number of women affected, pelvic floor pain and dysfunction are commonly overlooked in women seeking medical care. Physiatrists are uniquely qualified to manage these patients because of their knowledge of the musculoskeletal and nervous systems and their awareness of the relationships among pain, physiology, and function. When evaluating women who have pelvic pain, practitioners must ask questions about history of urinary or fecal incontinence, dyspareunia, or pelvic pain with certain activities or associated with menses, surgery, or trauma. If left unidentified, pelvic floor dysfunction can deter individuals from normal bowel and bladder function, intimacy, and even engagement in work and social functions. This article introduces pelvic floor anatomy, neurophysiology, and function and provides an overview of pelvic pain and pelvic floor dysfunctions and their recognition and treatment.

  2. Recruiting Women Smokers: The Engineering of Consent

    OpenAIRE

    Brandt, Allan M.

    1996-01-01

    A range of social forces contributed to the effective recruitment of women to cigarette smoking in the crucial period between 1900 and 1940. Cigarette advertisers and public relations experts recognized the significance of womens changing roles and the rising culture of consumption, and worked to create specific meanings for the cigarette to make it appeal to women. The cigarette was a flexible symbol, with a remarkably elastic set of meanings; for women, it represented rebellious independenc...

  3. Analysis of Gait Pattern to Recognize the Human Activities

    Directory of Open Access Journals (Sweden)

    Jay Prakash Gupta

    2014-09-01

    Full Text Available Human activity recognition based on the computer vision is the process of labelling image sequences with action labels. Accurate systems for this problem are applied in areas such as visual surveillance, human computer interaction and video retrieval. The challenges are due to variations in motion, recording settings and gait differences. Here we propose an approach to recognize the human activities through gait. Activity recognition through Gait is the process of identifying an activity by the manner in which they walk. The identification of human activities in a video, such as a person is walking, running, jumping, jogging etc are important activities in video surveillance. We contribute the use of Model based approach for activity recognition with the help of movement of legs only. Experimental results suggest that our method are able to recognize the human activities with a good accuracy rate and robust to shadows present in the videos.

  4. Discovering Activities to Recognize and Track in a Smart Environment.

    Science.gov (United States)

    Rashidi, Parisa; Cook, Diane J; Holder, Lawrence B; Schmitter-Edgecombe, Maureen

    2011-01-01

    The machine learning and pervasive sensing technologies found in smart homes offer unprecedented opportunities for providing health monitoring and assistance to individuals experiencing difficulties living independently at home. In order to monitor the functional health of smart home residents, we need to design technologies that recognize and track activities that people normally perform as part of their daily routines. Although approaches do exist for recognizing activities, the approaches are applied to activities that have been pre-selected and for which labeled training data is available. In contrast, we introduce an automated approach to activity tracking that identifies frequent activities that naturally occur in an individual's routine. With this capability we can then track the occurrence of regular activities to monitor functional health and to detect changes in an individual's patterns and lifestyle. In this paper we describe our activity mining and tracking approach and validate our algorithms on data collected in physical smart environments.

  5. Federally-Recognized Tribes of the Columbia-Snake Basin.

    Energy Technology Data Exchange (ETDEWEB)

    United States. Bonneville Power Administration

    1997-11-01

    This is an omnibus publication about the federally-recognized Indian tribes of the Columbia-Snake river basin, as presented by themselves. It showcases several figurative and literal snapshots of each tribe, bits and pieces of each tribe`s story. Each individual tribe or tribal confederation either submitted its own section to this publication, or developed its own section with the assistance of the writer-editor. A federally-recognized tribe is an individual Indian group, or confederation of Indian groups, officially acknowledged by the US government for purposes of legislation, consultation and benefits. This publication is designed to be used both as a resource and as an introduction to the tribes. Taken together, the sections present a rich picture of regional indian culture and history, as told by the tribes.

  6. Recognizing and treating toilet-seat contact dermatitis in children.

    Science.gov (United States)

    Litvinov, Ivan V; Sugathan, Paramoo; Cohen, Bernard A

    2010-02-01

    Toilet-seat contact dermatitis is a common condition around the world and is reemerging in the United States. It can be easily recognized and treated. However, few practitioners consider this diagnosis, which results in a delay in treatment and often exacerbation of the skin eruption. In the past, exposure to wooden toilet seats and associated varnish, lacquers, and paints led to the development of an allergic contact dermatitis on the buttocks and posterior thighs. In recent years, most public facilities have changed to plastic seats, resulting in a change in the clinical presentation of toilet-seat dermatitis. We present 5 cases of toilet-seat dermatitis in children from the United States and India and review the history, presentation, and clinical course of the disease. Our findings suggest that toilet-seat dermatitis is more common than previously recognized and should be considered in any child with a dermatitis that involves the buttocks and posterior thighs.

  7. T cells recognizing a peptide contaminant undetectable by mass spectrometry

    DEFF Research Database (Denmark)

    Brezar, Vedran; Culina, Slobodan; Østerbye, Thomas

    2011-01-01

    Synthetic peptides are widely used in immunological research as epitopes to stimulate their cognate T cells. These preparations are never completely pure, but trace contaminants are commonly revealed by mass spectrometry quality controls. In an effort to characterize novel major histocompatibility...... complex (MHC) Class I-restricted ß-cell epitopes in non-obese diabetic (NOD) mice, we identified islet-infiltrating CD8+ T cells recognizing a contaminating peptide. The amount of this contaminant was so small to be undetectable by direct mass spectrometry. Only after concentration by liquid...... chromatography, we observed a mass peak corresponding to an immunodominant islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP)(206-214) epitope described in the literature. Generation of CD8+ T-cell clones recognizing IGRP(206-214) using a novel method confirmed the identity...

  8. Recognizing of rights of mentally disabled individuals and school integration.

    OpenAIRE

    CZINEGOVÁ, Eva

    2009-01-01

    The attitude of society towards persons with mental disabilities and recognizing their rights and dignity are important evaluation aspects of the society. The Czech Republic follows modern European trends that provide the mentally disabled with the same extent of activities as people without disabilities. As a topic of my thesis I have chosen the mentally disabled children´s right to education. The thesis addresses attitudes of primary school headmasters as well as attitudes of the staff of s...

  9. [Studies on selectivity of recognizing factor for rmf promoter].

    Science.gov (United States)

    Ding, Q; Akira, I

    1997-02-01

    The truncted DNA template carrying rmf gene promoter was transcripted by E. coli RNA polymorase holoenzyme (E sigma) reconstituted with core enzyme (alpha 2 beta beta') and sigma 70 or sigma 38 in vitro. The initional site of the transcription of rmf was confirmed with different restriction endonuclease. rmf promoter can be recognized by E sigma 70 but not E sigma 38. The suitable temperature for in vitro transcription was 37 degrees C, NaCl concentration was 50 mmol/L.

  10. Meningococcemia: recognizing and reducing complications in pediatric patients.

    Science.gov (United States)

    Carno, M A

    1994-08-01

    Meningococcemia is a true infectious emergency that requires tremendous skill and collaboration among health-care professionals to reduce the high morbidity and mortality associated with this disease. Complications and sequelae may effect virtually every body system with meningococcal disease. The critical care nurse plays a crucial role in preventing and recognizing complications to reduce serious consequences, including respiratory distress syndrome, myocarditis, cardiovascular collapse, coagulopathies, major skin loss, and limb amputations.

  11. Characterization of CTL Recognized Epitopes on Human Breast Tumors

    Science.gov (United States)

    1996-09-01

    angina ], or uncontrolled arrhythmias). 18. Patients who have an intercurrent illness or chronically taking medication (e.g. antihistamines) which would...Nonispecifc T wave Asymnptomatic ST Angina without Acute myocardial ischentia flattening and T wave changes evidence for infairction Infarction suggesting...M, Jacobson L, Vincent A, Newsom-Davis J, and Willcox N: Peptide-selected T cell lines from mysastenia gravis patients and controls recognize epitopes

  12. Polyamine transporter recognization and antitumor effects of anthracenymethyl homospermidine.

    Science.gov (United States)

    Xie, Song-Qiang; Wang, Jian-Hong; Ma, Hong-Xia; Cheng, Peng-Fei; Zhao, Jin; Wang, Chao-Jie

    2009-09-19

    This study was conducted to examine the polyamine transporter (PAT) recognization and antitumor effects of anthracenymethyl homospermidine (ANTMHspd) and its apoptotic mechanism in B16 melanoma cells. ANTMHspd promoted a dose-dependent apoptosis in B16 melanoma cells and the apoptosis was associated with the excellent PAT recognization, externalization of cell membrane phosphatidylserine and the disruption of mitochondria, these processes were correlated with up-regulation of polyamine oxidase, an increase in intracellular reactive oxygen species (ROS) production and oxidative stress. In addition, reduction of MMP, release of cytochrome c, up-regulation of Bax protein expression, down-regulation of Bcl-2 protein expression, and activation of caspase-3, caspase-9 were also observed in B16 cells after treatment with ANTMHspd. Furthermore, N-acetylcysteine obviously antagonized ANTMHspd-induced apoptosis. Importantly, ANTMHspd was found to be better tolerated and revealed potent antitumor effect on inhibiting tumor growth in situ and suppressing pulmonary metastasis in xenograft mice model. These data demonstrate that ANTMHspd is an excellent PAT recognization and potent antitumor agent.

  13. Recognizing surgeon's actions during suture operations from video sequences

    Science.gov (United States)

    Li, Ye; Ohya, Jun; Chiba, Toshio; Xu, Rong; Yamashita, Hiromasa

    2014-03-01

    Because of the shortage of nurses in the world, the realization of a robotic nurse that can support surgeries autonomously is very important. More specifically, the robotic nurse should be able to autonomously recognize different situations of surgeries so that the robotic nurse can pass necessary surgical tools to the medical doctors in a timely manner. This paper proposes and explores methods that can classify suture and tying actions during suture operations from the video sequence that observes the surgery scene that includes the surgeon's hands. First, the proposed method uses skin pixel detection and foreground extraction to detect the hand area. Then, interest points are randomly chosen from the hand area so that their 3D SIFT descriptors are computed. A word vocabulary is built by applying hierarchical K-means to these descriptors, and the words' frequency histogram, which corresponds to the feature space, is computed. Finally, to classify the actions, either SVM (Support Vector Machine), Nearest Neighbor rule (NN) for the feature space or a method that combines "sliding window" with NN is performed. We collect 53 suture videos and 53 tying videos to build the training set and to test the proposed method experimentally. It turns out that the NN gives higher than 90% accuracies, which are better recognition than SVM. Negative actions, which are different from either suture or tying action, are recognized with quite good accuracies, while "Sliding window" did not show significant improvements for suture and tying and cannot recognize negative actions.

  14. Smart responsive microcapsules capable of recognizing heavy metal ions.

    Science.gov (United States)

    Pi, Shuo-Wei; Ju, Xiao-Jie; Wu, Han-Guang; Xie, Rui; Chu, Liang-Yin

    2010-09-15

    Smart responsive microcapsules capable of recognizing heavy metal ions are successfully prepared with oil-in-water-in-oil double emulsions as templates for polymerization in this study. The microcapsules are featured with thin poly(N-isopropylacrylamide-co-benzo-18-crown-6-acrylamide) (P(NIPAM-co-BCAm)) membranes, and they can selectively recognize special heavy metal ions such as barium(II) or lead(II) ions very well due to the "host-guest" complexation between the BCAm receptors and barium(II) or lead(II) ions. The stable BCAm/Ba(2+) or BCAm/Pb(2+) complexes in the P(NIPAM-co-BCAm) membrane cause a positive shift of the volume phase transition temperature of the crosslinked P(NIPAM-co-BCAm) hydrogel to a higher temperature, and the repulsion among the charged BCAm/Ba(2+) or BCAm/Pb(2+) complexes and the osmotic pressure within the P(NIPAM-co-BCAm) membranes result in the swelling of microcapsules. Induced by recognizing barium(II) or lead(II) ions, the prepared microcapsules with P(NIPAM-co-BCAm) membranes exhibit isothermal and significant swelling not only in outer and inner diameters but also in the membrane thickness. The proposed microcapsules in this study are highly attractive for developing smart sensors and/or carriers for detection and/or elimination of heavy metal ions. Copyright 2010 Elsevier Inc. All rights reserved.

  15. Women's Health

    Science.gov (United States)

    Women have unique health issues. And some of the health issues that affect both men and women can affect women differently. Unique issues ... and men also have many of the same health problems. But these problems can affect women differently. ...

  16. Women and Agriculture: Blending the Facts with Fiction.

    Science.gov (United States)

    Morris, Gayle A.

    Though the diversity of farm women's contributions was recognized by novelists such as Cather, Aldrich, and Sandoz in the 1800s, it would be many decades later before the work efforts of farm women would be formally recognized by the government and agricultural researchers. The West used a masculine environment, and farm animals and females where…

  17. Proteomics beyond trypsin

    NARCIS (Netherlands)

    Tsiatsiani, Liana|info:eu-repo/dai/nl/380468506; Heck, Albert J R|info:eu-repo/dai/nl/105189332

    2015-01-01

    Peptide-centered shotgun analysis of proteins has been the core technology in mass spectrometry based proteomics and has enabled numerous biological discoveries, such as the large-scale charting of protein-protein interaction networks, the quantitative analysis of protein post-translational

  18. Effect of trypsin inhibitor activity in soya bean on growth performance, protein digestibility and incidence of sub-clinical necrotic enteritis in broiler chicken flocks.

    Science.gov (United States)

    Palliyeguru, M W C D; Rose, S P; Mackenzie, A M

    2011-06-01

    1. The effect of three different levels of dietary trypsin inhibitor activity (achieved by varying the amount of non-toasted full fat soya bean in replacement for toasted full fat soya bean) on the incidence of spontaneously-occurring sub-clinical necrotic enteritis (NE) in broiler chickens was compared. A fourth dietary treatment compared the effect of a diet that used potato protein concentrate as the major protein source. The determined trypsin inhibitor activity increased with the increasing content of non-toasted soya bean: 1·90, 6·21, 8·46 and 3·72 mg/g for the three soya bean diets (0, 100 and 200 g of non-toasted soya bean/kg) and the potato protein diet respectively. 2. Although increasing amounts of the non-toasted full-fat soya bean increased the feed intakes of the birds, there was a marked reduction in protein digestibility, weight gain and feed conversion efficiency. 3. There was a linear increase in sub-clinical NE lesions in the duodenum, jejunum, mid small intestine and ileum with increasing non-toasted soya bean. Caecal Clostridium perfringens counts increased with the increasing dietary content of non-toasted soya bean. Serum α-toxin antibodies were higher in the birds fed the 200 g non-toasted soya bean/kg diet compared with the other diets. 4. The results demonstrated that variation in the amount of non-toasted dietary soya bean not only affects growth performance of broilers but also affects the incidence of sub-clinical necrotic enteritis in the flock. Ensuring the lowest possible trypsin-inhibitor activity in soya bean samples is a valuable tool to improve the health and welfare of birds and in reducing the financial losses from this disease.

  19. A novel discriminant score based on tumor-associated trypsin inhibitor for accurate diagnosis of metastasis in patients with breast cancer.

    Science.gov (United States)

    El-mezayen, Hatem A; Metwally, Fatheya M; Darwish, Hossam

    2014-03-01

    Invasion and metastasis of solid tumors require proteolytic enzymes for degradation of the basal membrane and extracellular matrix. Currently, there are no reliable methodologies to predict the risk for metastatic disease. In this context, our aim has been focused on the development of a noninvasive score based on tumor-associated trypsin inhibitor (TATI) for the assessment of metastasis in patients with breast cancer. TATI, trypsin, and soluble epidermal growth factor receptor (sEGFR) were assayed by enzyme-linked immunosorbent assay. CA 15.3 serum level was assayed by microparticle enzyme immunoassay in 265 patients with breast cancer. Statistical analyses were performed by logistic regression and receiver operating characteristic analysis curves. Using multivariate discriminant analysis, a score is selected based on absolute values of the four biochemical markers: TATI-metastatic breast cancer score (TATI-MBCS) = [0.03 × CA 15.3 (U/L) + 0.039 × TATI (ng/ml) + 0.04 × trypsin (ng/ml) + 0.023 × sEGFR (ng/ml) - 6.49 (numerical constant)]. This function correctly classified 84% of metastatic breast cancer at cutoff value = 0.62 (i.e., greater than 0.62 indicates patients with metastatic breast cancer and less than 0.62 indicates patients with nonmetastatic breast cancer). In conclusion, TATI-MBCS is a novel, noninvasive, and simple score which can be applied to discriminate patients with metastatic breast cancer.

  20. Molecular Basis of Enhanced Activity in Factor VIIa-Trypsin Variants Conveys Insights into Tissue Factor-mediated Allosteric Regulation of Factor VIIa Activity

    DEFF Research Database (Denmark)

    Sorensen, Anders B.; Madsen, Jesper Jonasson; Svensson, L. Anders

    2016-01-01

    -ray crystallography, we show that the introduced 170 loop from trypsin directly interacts with the FVIIa active site, stabilizing segment 215-217 and activation loop 3, leading to enhanced activity. Molecular dynamics simulations and novel fluorescence quenching studies support that segment 215-217 conformation...... is pivotal to the enhanced activity of the FVIIa variants. We speculate that the allosteric regulation of FVIIa activity by TF binding follows a similar path in conjunction with protease domain N terminus insertion, suggesting a more complete molecular basis of TF-mediated allosteric enhancement of FVIIa...

  1. Women in Meteorology.

    Science.gov (United States)

    Lemone, Margaret A.; Waukau, Patricia L.

    1982-11-01

    The names of 927 women who are or have been active in meteorology or closely related fields have been obtained from various sources. Of these women, at least 500 are presently active. An estimated 4-5% of the total number of Ph.D.s in meteorology are awarded to women. About 10% of those receiving B.S. and M.S. degrees are women.The work patterns, accomplishments, and salaries of employed women meteorologists have been summarized from 330 responses to questionnaires, as functions of age, family status, part- or full-time working status, and employing institutions. It was found that women meteorologists holding Ph.D.s are more likely than their male counterparts to be employed by universities. As increasing number of women were employed in operational meteorology, although few of them were married and fewer still responsible for children. Several women were employed by private industry and some had advanced into managerial positions, although at the present time, such positions remain out of the reach of most women.The subjective and objective effects of several gender-related factors have been summarized from the comments and responses to the questionnaires. The primary obstacles to advancement were found to be part-time work and the responsibility for children. Part-time work was found to have a clearly negative effect on salary increase as a function of age. prejudicated discrimination and rules negatively affecting women remain important, especially to the older women, and affirmative action programs are generally seen as beneficial.Surprisingly, in contrast to the experience of women in other fields of science, women Ph.D.s in meteorology earn salaries comparable of their employment in government or large corporations and universities where there are strong affirmative action programs and above-average salaries. Based on the responses to the questionnaire, the small size of the meteorological community is also a factor, enabling women to become recognized

  2. Procrastination in Recognizing the Rights of Domestic Workers

    African Journals Online (AJOL)

    Mussie_Mezgebo

    United States” The Association for Women's Rights in Development (AWID), Toronto, p. 5. 21 Kidist Mulugeta (2012), Vulnerability, Legal Protection and Work Conditions of. Domestic Workers in Addis Ababa (The Hague: International Institute of Social Studies. Research Paper), p. 11. 22 Andrea Cristina Mercado and Ai-jen ...

  3. Recognizing and Preventing Overexposure to Methylmercury from Fish and Seafood Consumption: Information for Physicians

    Directory of Open Access Journals (Sweden)

    Susan M. Silbernagel

    2011-01-01

    Full Text Available Fish is a valuable source of nutrition, and many people would benefit from eating fish regularly. But some people eat a lot of fish, every day or several meals per week, and thus can run a significant risk of overexposure to methylmercury. Current advice regarding methylmercury from fish consumption is targeted to protect the developing brain and nervous system but adverse health effects are increasingly associated with adult chronic low-level methylmercury exposure. Manifestations of methylmercury poisoning are variable and may be difficult to detect unless one considers this specific diagnosis and does an appropriate test (blood or hair analysis. We provide information to physicians to recognize and prevent overexposure to methylmercury from fish and seafood consumption. Physicians are urged to ask patients if they eat fish: how often, how much, and what kinds. People who eat fish frequently (once a week or more often and pregnant women are advised to choose low mercury fish.

  4. Engineering support vector machine kernels that recognize translation initiation sites.

    Science.gov (United States)

    Zien, A; Rätsch, G; Mika, S; Schölkopf, B; Lengauer, T; Müller, K R

    2000-09-01

    In order to extract protein sequences from nucleotide sequences, it is an important step to recognize points at which regions start that code for proteins. These points are called translation initiation sites (TIS). The task of finding TIS can be modeled as a classification problem. We demonstrate the applicability of support vector machines for this task, and show how to incorporate prior biological knowledge by engineering an appropriate kernel function. With the described techniques the recognition performance can be improved by 26% over leading existing approaches. We provide evidence that existing related methods (e.g. ESTScan) could profit from advanced TIS recognition.

  5. New efficient algorithm for recognizing handwritten Hindi digits

    Science.gov (United States)

    El-Sonbaty, Yasser; Ismail, Mohammed A.; Karoui, Kamal

    2001-12-01

    In this paper a new algorithm for recognizing handwritten Hindi digits is proposed. The proposed algorithm is based on using the topological characteristics combined with statistical properties of the given digits in order to extract a set of features that can be used in the process of digit classification. 10,000 handwritten digits are used in the experimental results. 1100 digits are used for training and another 5500 unseen digits are used for testing. The recognition rate has reached 97.56%, a substitution rate of 1.822%, and a rejection rate of 0.618%.

  6. Recognizing the Stranger: Recognition Scenes in the Gospel of John

    DEFF Research Database (Denmark)

    Larsen, Kasper Bro

    Recognizing the Stranger is the first monographic study of recognition scenes and motifs in the Gospel of John. The recognition type-scene (anagnōrisis) was a common feature in ancient drama and narrative, highly valued by Aristotle as a touching moment of truth, e.g., in Oedipus’ tragic self......-discovery and Odysseus’ happy homecoming. The book offers a reconstruction of the conventions of the genre and argues that it is one of the most recurrent and significant literary forms in the Gospel. When portraying Jesus as the divine stranger from heaven, the Gospel employs and transforms the formal and ideological...

  7. Recognizing and Treating Malaria in U.S. Residents

    Centers for Disease Control (CDC) Podcasts

    2010-06-09

    This podcast is an overview of the Clinician Outreach and Communication Activity (COCA) Call: It's a Small World After All: Dengue and Malaria in U.S. Residents - Recognizing and Treating These Mosquito-borne Diseases. CDC's David Townes discusses clinical presentation, transmission, prevention strategies, new treatments, and malaria resources available to health care providers.  Created: 6/9/2010 by Division of Parasitic Diseases and Malaria, Center for Global Health and Emergency Communication System (ECS)/Joint Information Center (JIC); Office of Public Health Preparedness and Response (OPHPR).   Date Released: 6/15/2010.

  8. Protective effect of Carbopol on enzymatic degradation of a peptide-like substrate. I: Effect of various concentrations and grades of Carbopol and other reaction variables on trypsin activity.

    Science.gov (United States)

    Vaidya, A P; Wigent, R J; Moore, J C; Schwartz, J B

    2007-01-01

    The purpose of this study was to determine and compare the effect of various concentrations and grades of Carbopol on trypsin-induced degradation of a prototype substrate, N(alpha)-benzoyl-L-arginine ethyl ester hydrochloride (BAEE). Effect of other reaction variables, such as viscosity and ionic strength of the medium on the trypsin activity, was also analyzed simultaneously. Four concentrations and three commercially available grades of Carbopol were used. The effect of Carbopol was expressed in terms of change in the velocity of degradation reaction. A modified trypsin assay was developed and used for analysis. Up to a concentration of 0.35% w/v, Carbopol 934P showed a concentration-dependent increase in its ability to reduce the rate of enzymatic hydrolysis of BAEE. Similar inhibitory effect was observed with all three grades of Carbopol. The activity of trypsin was unaffected by other reaction variables, suggesting that interaction between the protein and the polymer could be the mechanism responsible for reduced trypsin activity. This study suggests that Carbopol can be a useful excipient for oral delivery of bioactive proteins and peptides, due to its ability to reduce the enzyme-induced degradation of these agents.

  9. Recognizing Induced Emotions of Happiness and Sadness from Dance Movement

    Science.gov (United States)

    Van Dyck, Edith; Vansteenkiste, Pieter; Lenoir, Matthieu; Lesaffre, Micheline; Leman, Marc

    2014-01-01

    Recent research revealed that emotional content can be successfully decoded from human dance movement. Most previous studies made use of videos of actors or dancers portraying emotions through choreography. The current study applies emotion induction techniques and free movement in order to examine the recognition of emotional content from dance. Observers (N = 30) watched a set of silent videos showing depersonalized avatars of dancers moving to an emotionally neutral musical stimulus after emotions of either sadness or happiness had been induced. Each of the video clips consisted of two dance performances which were presented side-by-side and were played simultaneously; one of a dancer in the happy condition and one of the same individual in the sad condition. After every film clip, the observers were asked to make forced-choices concerning the emotional state of the dancer. Results revealed that observers were able to identify the emotional state of the dancers with a high degree of accuracy. Moreover, emotions were more often recognized for female dancers than for their male counterparts. In addition, the results of eye tracking measurements unveiled that observers primarily focus on movements of the chest when decoding emotional information from dance movement. The findings of our study show that not merely portrayed emotions, but also induced emotions can be successfully recognized from free dance movement. PMID:24587026

  10. Recognizing induced emotions of happiness and sadness from dance movement.

    Directory of Open Access Journals (Sweden)

    Edith Van Dyck

    Full Text Available Recent research revealed that emotional content can be successfully decoded from human dance movement. Most previous studies made use of videos of actors or dancers portraying emotions through choreography. The current study applies emotion induction techniques and free movement in order to examine the recognition of emotional content from dance. Observers (N = 30 watched a set of silent videos showing depersonalized avatars of dancers moving to an emotionally neutral musical stimulus after emotions of either sadness or happiness had been induced. Each of the video clips consisted of two dance performances which were presented side-by-side and were played simultaneously; one of a dancer in the happy condition and one of the same individual in the sad condition. After every film clip, the observers were asked to make forced-choices concerning the emotional state of the dancer. Results revealed that observers were able to identify the emotional state of the dancers with a high degree of accuracy. Moreover, emotions were more often recognized for female dancers than for their male counterparts. In addition, the results of eye tracking measurements unveiled that observers primarily focus on movements of the chest when decoding emotional information from dance movement. The findings of our study show that not merely portrayed emotions, but also induced emotions can be successfully recognized from free dance movement.

  11. Recognizing young children with high potential: U-STARS∼PLUS.

    Science.gov (United States)

    Coleman, Mary Ruth

    2016-08-01

    Hands-on science is the ideal platform for observing young children's ability to solve problems, think deeply, and use their creative ingenuity to explore the world around them. Science is naturally interesting and offers authentic reasons to read for information and use math skills to collect, compile, and analyze data. This chapter will share one approach to nurturing and recognizing young children with high-potential: U-STARS∼PLUS (Using Science, Talents, and Abilities to Recognize Students∼Promoting Learning for Underrepresented Students). Each of the five components (high-end learning environments; teacher's observations of potential; engaging science activities; partnerships with parents; and capacity building for system change) will be explained. Concrete examples will be given for each area showing how it works and why it is important. Special attention will be paid to the needs of educationally vulnerable gifted children who remain underserved: racially, ethnically, and linguistically different; economically disadvantaged, and children who are twice exceptional (2e). © 2016 New York Academy of Sciences.

  12. Recognizing induced emotions of happiness and sadness from dance movement.

    Science.gov (United States)

    Van Dyck, Edith; Vansteenkiste, Pieter; Lenoir, Matthieu; Lesaffre, Micheline; Leman, Marc

    2014-01-01

    Recent research revealed that emotional content can be successfully decoded from human dance movement. Most previous studies made use of videos of actors or dancers portraying emotions through choreography. The current study applies emotion induction techniques and free movement in order to examine the recognition of emotional content from dance. Observers (N = 30) watched a set of silent videos showing depersonalized avatars of dancers moving to an emotionally neutral musical stimulus after emotions of either sadness or happiness had been induced. Each of the video clips consisted of two dance performances which were presented side-by-side and were played simultaneously; one of a dancer in the happy condition and one of the same individual in the sad condition. After every film clip, the observers were asked to make forced-choices concerning the emotional state of the dancer. Results revealed that observers were able to identify the emotional state of the dancers with a high degree of accuracy. Moreover, emotions were more often recognized for female dancers than for their male counterparts. In addition, the results of eye tracking measurements unveiled that observers primarily focus on movements of the chest when decoding emotional information from dance movement. The findings of our study show that not merely portrayed emotions, but also induced emotions can be successfully recognized from free dance movement.

  13. Effects of Facial Expressions on Recognizing Emotions in Dance Movements

    Directory of Open Access Journals (Sweden)

    Nao Shikanai

    2011-10-01

    Full Text Available Effects of facial expressions on recognizing emotions expressed in dance movements were investigated. Dancers expressed three emotions: joy, sadness, and anger through dance movements. We used digital video cameras and a 3D motion capturing system to record and capture the movements. We then created full-video displays with an expressive face, full-video displays with an unexpressive face, stick figure displays (no face, or point-light displays (no face from these data using 3D animation software. To make point-light displays, 13 markers were attached to the body of each dancer. We examined how accurately observers were able to identify the expression that the dancers intended to create through their dance movements. Dance experienced and inexperienced observers participated in the experiment. They watched the movements and rated the compatibility of each emotion with each movement on a 5-point Likert scale. The results indicated that both experienced and inexperienced observers could identify all the emotions that dancers intended to express. Identification scores for dance movements with an expressive face were higher than for other expressions. This finding indicates that facial expressions affect the identification of emotions in dance movements, whereas only bodily expressions provide sufficient information to recognize emotions.

  14. On learning to recognize spectrally reduced speech: II. Individual differences

    Science.gov (United States)

    Chiu, Peter; Eng, Michelle; Strange, Bethany; Yampolsky, Sasha; Waters, Gloria

    2002-05-01

    When patients with cochlear implants attempt speech recognition, or when listeners with normal hearing try to recognize spectrally reduced speech, performance may suffer because of additional demands on cognitive resources imposed by sensory degradation. Some previous studies have reported a significant correlation between digit span and recognition of spectrally reduced speech, but others have not [Eisenberg et al., J. Acoust. Soc. Am. 107, 2704-2710 (2000)]. In this study, we administered 3 separate measures of working memory capacity (i.e., digit span, alphabet span, and sentence span) and 1 measure of reading ability (American National Adult Reading Test, ANART) to normal-hearing listeners who participated in our previous studies [Chiu et al., J. Acoust. Soc. Am. 109, 2501 (2001) and Fishbeck and Chiu, J. Acoust. Soc. Am. 109, 2504 (2001)]. Preliminary data show that the listener's ability to recognize spectrally reduced speech shows the strongest relationship with ANART scores and the weakest relationship with digit span. Implications of the current findings on theories of speech recognition and working memory will be discussed. [Work supported by URC, Univ. of Cincinnati.

  15. Recognizing age-separated face images: humans and machines.

    Science.gov (United States)

    Yadav, Daksha; Singh, Richa; Vatsa, Mayank; Noore, Afzel

    2014-01-01

    Humans utilize facial appearance, gender, expression, aging pattern, and other ancillary information to recognize individuals. It is interesting to observe how humans perceive facial age. Analyzing these properties can help in understanding the phenomenon of facial aging and incorporating the findings can help in designing effective algorithms. Such a study has two components--facial age estimation and age-separated face recognition. Age estimation involves predicting the age of an individual given his/her facial image. On the other hand, age-separated face recognition consists of recognizing an individual given his/her age-separated images. In this research, we investigate which facial cues are utilized by humans for estimating the age of people belonging to various age groups along with analyzing the effect of one's gender, age, and ethnicity on age estimation skills. We also analyze how various facial regions such as binocular and mouth regions influence age estimation and recognition capabilities. Finally, we propose an age-invariant face recognition algorithm that incorporates the knowledge learned from these observations. Key observations of our research are: (1) the age group of newborns and toddlers is easiest to estimate, (2) gender and ethnicity do not affect the judgment of age group estimation, (3) face as a global feature, is essential to achieve good performance in age-separated face recognition, and (4) the proposed algorithm yields improved recognition performance compared to existing algorithms and also outperforms a commercial system in the young image as probe scenario.

  16. Patient Selection in Plastic Surgery: Recognizing Body Dysmorphic Disorder

    Directory of Open Access Journals (Sweden)

    Cihan Sahin

    2013-04-01

    Full Text Available Plastic surgery is a branch of medicine that provides significant improvements to the people with positive changes. But first of all, this branch has a characteristic which requires analysing patients' psychological situation very carefully. Plastic surgeons are often confronted by patients with mental disorders seeking aesthetic surgery. It is imperative for surgeons to recognize possible underlying psychiatric illnesses. Common psychiatric conditions seen in cosmetic surgery patients include body dysmorphic disorder (BDD, narcissistic personality disorder and histrionic personality disorders. BDD is of particular importance to plastic surgeons. Because outrageous dissatisfaction with one's appearance may conceal psychopathologic traits that are not always easily recognizable, and which, if neglected, may result in serious iatrogenic and medicolegal consequences, we hope that this paper will help plastic surgeons in ultimately preventing patient and surgeon dissatisfaction within the population of patients with psychiatric disorders, and should recognize the diagnostic features of body dysmorphic disorder and screen psychologically unstable patients who may never be satisfied with surgery. [Arch Clin Exp Surg 2013; 2(2.000: 109-115

  17. Exploring the recognized bio-mimicry materials for gas sensing.

    Science.gov (United States)

    Wu, T Z; Lo, Y R; Chan, E C

    2001-12-01

    This study was undertaken to synthesize peptides that are partially similar to the binding sites of human olfactory receptor protein. First, a putative 3-D model structure of human olfactory receptor protein (P30953) was modeled using a molecular simulation method. The computer docking simulation was then performed to determine the most plausible binding sites between the model structure and target gases, trimethylamine, ammonia, acetic acid, and o-xylene. According to the simulation result, a series of polypeptide sequences, horp61 for TMA, horp103 for o-xylene, horp109 for ammonia, and horp193 for acetic acid as recognized molecules were designed for gas sensing purposes. Preparing these peptides as corresponding gas sensing probes, the results showed a high relative sensitivity response of 6.7 for TMA (probe horp61), 5.1 for o-xylene (probe horp103), 11 for ammonia (probe horp109), and 28 for acetic acid (probe horp193), respectively. These results indicate that peptide mimicking of binding domain on olfactory receptor opens a new window and offers a novel strategy for the further development of recognized materials for gas sensing.

  18. Screening of serine protease inhibitors with antimicrobial activity using iron oxide nanoparticles functionalized with dextran conjugated trypsin and in silico analyses of bacterial serine protease inhibition.

    Science.gov (United States)

    Mandal, Santi M; Porto, William F; De, Debasis; Phule, Ajit; Korpole, Suresh; Ghosh, Ananta K; Roy, Sanat K; Franco, Octavio L

    2014-01-21

    Plants produce a variety of proteins and peptides which are involved in their defense against pathogens. Serine protease inhibitors are a well-established class of inhibitors correlated with plant defense. Increased levels of protease inhibitors delay cell damage and expand the cell's life-span. Recently, the rapid emergence of antibiotic-resistant microbial pathogens has prompted immense interest in purifying novel antimicrobial proteins or peptides from plant sources. Usually, the purification of protease inhibitors is accomplished by salt-extraction, ultrafiltration and affinity chromatography. Here, we developed a novel approach based on iron oxide nanoparticles conjugated to dextran functionalized with trypsin beads that accelerate the quick screening and purification of antimicrobial peptides with serine protease inhibitor activity. The method described here also works for screening other inhibitors using particular protein kinases, and it is therefore a novel tool for use as the leading method in the development of novel antimicrobial agents with protease inhibitory activity. Finally, and no less important, molecular modelling and dynamics studies of a homologous inhibitor studied here with Escherichia coli trypsin and chymotrypsin are provided in order to shed some light on inhibitor-enzyme interactions.

  19. Identification and characterization of a serine protease inhibitor with two trypsin inhibitor-like domains from the human hookworm Ancylostoma duodenale.

    Science.gov (United States)

    Jin, Xian; Deng, Li; Li, Hui; Zhang, Zhenlin; He, Qingfeng; Yang, Chen; Jiang, Hanguo; Zhu, Xing-Quan; Peng, Lifei

    2011-02-01

    Protease inhibitors play important roles in the parasitic nematodes' survival within their host, in the development and reproduction of the parasites. The present study described the isolation, identification, and characterization of a novel member of the Ascaris family of serine protease inhibitors, designated AduTIL-1, from the human hookworm Ancylostoma duodenale. AduTIL-1 is composed of a signal sequence and two trypsin inhibitor-like (TIL) domains, which showed the highest similarity with OdmCRP, a putative serine protease inhibitor with two TIL domains in Oesophagostomum dentatum. Each TIL domain of the AduTIL-1 was expressed in Escherichia coli, and their inhibitory activities against serine proteases from animals and human were characterized, respectively. Both of the two TIL domains inhibited human neutrophil elastase and pancreatic trypsin, but different in effectiveness. Although the first TIL domain of AduTIL-1 inhibited bovine pancreatic chymotrypsin (Ki=18.0 nM), both of the two domains showed no inhibitory activity against the human pancreatic chymotrypsin. Immunohistochemical studies demonstrated that AduTIL-1 was localized in esophagus, intestine, and cuticular surface of the adult worms. These results suggested that AduTIL-1 may be involved in the survival of A. duodenale in host by targeting related digestive enzymes and neutrophil elastase.

  20. Preparation of a novel polymer monolith with functional polymer brushes by two-step atom-transfer radical polymerization for trypsin immobilization.

    Science.gov (United States)

    Li, Nan; Zheng, Wei; Shen, Ying; Qi, Li; Li, Yaping; Qiao, Juan; Wang, Fuyi; Chen, Yi

    2014-12-01

    Novel porous polymer monoliths grafted with poly{oligo[(ethylene glycol) methacrylate]-co-glycidyl methacrylate} brushes were fabricated via two-step atom-transfer radical polymerization and used as a trypsin-based reactor in a continuous flow system. This is the first time that atom-transfer radical polymerization technique was utilized to design and construct polymer monolith bioreactor. The prepared monoliths possessed excellent permeability, providing fast mass transfer for enzymatic reaction. More importantly, surface properties, which were modulated via surface-initiated atom-transfer radical polymerization, were found to have a great effect on bioreactor activities based on Michaelis-Menten studies. Furthermore, three model proteins were digested by the monolith bioreactor to a larger degree within dramatically reduced time (50 s), about 900 times faster than that by free trypsin (12 h). The proposed method provided a platform to prepare porous monoliths with desired surface properties for immobilizing various enzymes. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Application and bioactive properties of CaTI, a trypsin inhibitor from Capsicum annuum seeds: membrane permeabilization, oxidative stress and intracellular target in phytopathogenic fungi cells.

    Science.gov (United States)

    Silva, Marciele S; Ribeiro, Suzanna Ff; Taveira, Gabriel B; Rodrigues, Rosana; Fernandes, Katia Vs; Carvalho, André O; Vasconcelos, Ilka Maria; Mello, Erica Oliveira; Gomes, Valdirene M

    2017-08-01

    During the last few years, a growing number of antimicrobial peptides have been isolated from plants and particularly from seeds. Recent results from our laboratory have shown the purification of a new trypsin inhibitor, named CaTI, from chilli pepper (Capsicum annuum L.) seeds. This study aims to evaluate the antifungal activity and mechanism of action of CaTI on phytopathogenic fungi and detect the presence of protease inhibitors in other species of this genus. Our results show that CaTI can inhibit the growth of the phytopathogenic fungi Colletotrichum gloeosporioides and C. lindemuthianum. CaTI can also permeabilize the membrane of all tested fungi. When testing the inhibitor on its ability to induce reactive oxygen species, an induction of reactive oxygen species (ROS) and nitric oxide (NO) particularly in Fusarium species was observed. Using CaTI coupled to fluorescein isothiocyanate (FITC), it was possible to determine the presence of the inhibitor inside the hyphae of the Fusarium oxysporum fungus. The search for protease inhibitors in other Capsicum species revealed their presence in all tested species. This paper shows the antifungal activity of protease inhibitors such as CaTI against phytopathogenic fungi. Antimicrobial peptides, among which the trypsin protease inhibitor family stands out, are present in different species of the genus Capsicum and are part of the chemical arsenal that plants use to defend themselves against pathogens. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  2. Effects of α,β-unsaturated lactones on larval survival and gut trypsin as well as oviposition response of Aedes aegypti.

    Science.gov (United States)

    Barros, Maria Ester S B; Freitas, Juliano C R; Santos, Geanne K N; da Silva, Rayane Cristine Santos; Pontual, Emmanuel V; Paiva, Patrícia M G; Napoleão, Thiago H; Navarro, Daniela M A F; Menezes, Paulo H

    2015-09-01

    Lactones are organic cyclic esters that have been described as larvicides against Aedes aegypti and as components of oviposition pheromone of Culex quinquefasciatus. This work describes the effect of six α,β-unsaturated lactones (5a-5f) on survival of A. aegypti fourth instar larvae (L4). It is also reported the effects of the lactones on L4 gut trypsin activity and oviposition behavior of A. aegypti females. Five lactones were able to kill L4 being the lactones 5a (LC50 of 39.05 ppm), 5e (LC50 of 36.30 ppm) and 5f (LC50 of 40.46 ppm) the most promising larvicides. Only the lactone 5a inhibited L4 gut trypsin activity, with an IC50 of 115.15 µg/mL. Lactones 5a, 5c, 5d and 5e did not exert deterrent or stimulatory effects on oviposition, whereas lactone 5b exhibited a strong deterrent oviposition activity. In conclusion, this work introduces new α,β-unsaturated lactones as promising alternatives to control A. aegypti dissemination. The larvicidal mechanism of the lactone 5a can involve the disruption of proteolysis at larval gut. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. Images of Women in Visual Culture

    Science.gov (United States)

    Lai, Alice

    2009-01-01

    Visual culture permeates women's everyday lives, affecting how they see themselves, the world, and their relationships with others. Increasing women's opportunities to recognize the power and problem of visual culture and subsequently to create new meanings through visual culture is an important teaching goal of the author. The author considered…

  4. Recognizing the value of assistance dogs in society.

    Science.gov (United States)

    Audrestch, Hilary M; Whelan, Chantelle T; Grice, David; Asher, Lucy; England, Gary C W; Freeman, Sarah L

    2015-10-01

    Assistance dogs are specially trained to undertake a variety of tasks to help individuals with disabilities. This review gives an overview of the different types of assistance dogs in the UK, including guide dogs, hearing dogs, mobility assistance dogs, service dogs and dual-purpose dogs. The literature describes many benefits of assistance dogs, including their impact on physical wellbeing and safety of their 'owners,' as well as on psychological wellbeing and social inclusion. The role of assistance dogs in society is widely recognized by the public, but is not currently acknowledged in government social policy. The current evidence on the benefits of assistance dogs is limited by the type and scale of current research. This article highlights the need for independent funding for high quality research to enable social care and policy makers to make evidence-based decisions on the value of assistance dogs to people with disabilities. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. Recognizing Dementia: Constructing Deconstruction in a Danish Memory Clinic.

    Science.gov (United States)

    Gjødsbøl, Iben M; Svendsen, Mette N

    2017-03-06

    This article investigates how a person with dementia is made up through intersubjective acts of recognition. Based on ethnographic fieldwork in a Danish memory clinic, we show that identification of disease requires patients to be substituted by their relatives in constructing believable medical narratives; yet during memory testing, patients are not allowed any substitution to clearly expose cognitive shortcomings. In combining works of theorists Ian Hacking and Paul Ricoeur, we argue that the clinical identification of dementia unmakes the knowing subject, a deconstruction that threatens to misrecognize and humiliate the person under examination. The article ends by proposing that dementia be the condition that forces us to rethink our ways of recognizing persons more generally. Thus, dementia diagnostics provide insights into different enactments of the person that invite us to explore practices of substitution and modes of interaction emerging when our fundamental dependency becomes unquestionable. © 2017 by the American Anthropological Association.

  6. Heterosexual identity and heterosexism: recognizing privilege to reduce prejudice.

    Science.gov (United States)

    Simoni, J M; Walters, K L

    2001-01-01

    We propose that heterosexuals' attitudes toward their own heterosexual identity evolve in a manner similar to the development of racial identity attitudes among Whites-another dominant group with privileged social status. A more developed heterosexual identity, we hypothesized, would be associated with diminished levels of negative attitudes toward non-heterosexuals. To test our hypotheses, we administered to 154 heterosexual students Herek's (1988) Attitudes Toward Gays and Lesbians scale measuring heterosexist (i.e., anti-gay) attitudes and a version of Helms and Carter's (1990) White Racial Identity Attitude Scale (WRIAS) modified to assess heterosexual identity attitudes. Hierarchical regression analyses controlling demographic indicators partially confirmed the hypothesized associations between evolving identity stages and less heterosexist attitudes. The role of recognizing one's social privilege is proposed (though not empirically examined) as a potential mediator between developing a dominant group identity and decreasing prejudicial attitudes towards non-privileged groups. Finally, implications and recommendations for future research and addressing heterosexism are presented.

  7. Serendipitous learning: Recognizing and fostering the potential of microblogging

    Directory of Open Access Journals (Sweden)

    Ilona Buchem

    2013-03-01

    Full Text Available This paper introduces the concept of serendipitous learning in the context of microblogging and discusses the potential of unplanned and unexpected discoveries for learning. Serendipitous learning as a subset of incidental learning refers to learning through gaining new insights, discovering unrevealed aspects and recognizing seemingly unrelated connections. This type of learning can occur by chance and as a by-product of other activities like information browsing through social status updates in microblogs. It is argued that engaging in microblogging in an open social network of users frequently generating new information enhances the possibilities of serendipitous discovery. The paper discusses possible factors facilitating serendipitous learning and concludes with recommendations for future research.

  8. Combined pulmonary fibrosis and emphysema: an increasingly recognized condition

    Directory of Open Access Journals (Sweden)

    Olívia Meira Dias

    2014-06-01

    Full Text Available Combined pulmonary fibrosis and emphysema (CPFE has been increasingly recognized in the literature. Patients with CPFE are usually heavy smokers or former smokers with concomitant lower lobe fibrosis and upper lobe emphysema on chest HRCT scans. They commonly present with severe breathlessness and low DLCO, despite spirometry showing relatively preserved lung volumes. Moderate to severe pulmonary arterial hypertension is common in such patients, who are also at an increased risk of developing lung cancer. Unfortunately, there is currently no effective treatment for CPFE. In this review, we discuss the current knowledge of the pathogenesis, clinical characteristics, and prognostic factors of CPFE. Given that most of the published data on CPFE are based on retrospective analysis, more studies are needed in order to address the role of emphysema and its subtypes; the progression of fibrosis/emphysema and its correlation with inflammation; treatment options; and prognosis.

  9. Random Deep Belief Networks for Recognizing Emotions from Speech Signals.

    Science.gov (United States)

    Wen, Guihua; Li, Huihui; Huang, Jubing; Li, Danyang; Xun, Eryang

    2017-01-01

    Now the human emotions can be recognized from speech signals using machine learning methods; however, they are challenged by the lower recognition accuracies in real applications due to lack of the rich representation ability. Deep belief networks (DBN) can automatically discover the multiple levels of representations in speech signals. To make full of its advantages, this paper presents an ensemble of random deep belief networks (RDBN) method for speech emotion recognition. It firstly extracts the low level features of the input speech signal and then applies them to construct lots of random subspaces. Each random subspace is then provided for DBN to yield the higher level features as the input of the classifier to output an emotion label. All outputted emotion labels are then fused through the majority voting to decide the final emotion label for the input speech signal. The conducted experimental results on benchmark speech emotion databases show that RDBN has better accuracy than the compared methods for speech emotion recognition.

  10. Truncated Autoinducing Peptide Conjugates Selectively Recognize and Kill Staphylococcus aureus.

    Science.gov (United States)

    Tsuchikama, Kyoji; Shimamoto, Yasuhiro; Anami, Yasuaki

    2017-06-09

    The accessory gene regulator (agr) of Staphylococcus aureus coordinates various pathogenic events and is recognized as a promising therapeutic target for virulence control. S. aureus utilizes autoinducing peptides (AIPs), cyclic-peptide signaling molecules, to mediate the agr system. Despite the high potency of synthetic AIP analogues in agr inhibition, the potential of AIP molecules as a delivery vehicle for antibacterial agents remains unexplored. Herein, we report that truncated AIP scaffolds can be fused with fluorophore and cytotoxic photosensitizer molecules without compromising their high agr inhibitory activity, binding affinity to the receptor AgrC, or cell specificity. Strikingly, a photosensitizer-AIP conjugate exhibited 16-fold greater efficacy in a S. aureus cell-killing assay than a nontargeting analogue. These findings highlight the potential of truncated AIP conjugates as useful chemical tools for in-depth biological studies and as effective anti-S. aureus agents.

  11. Understanding and recognizing the Pelger-Huët anomaly.

    Science.gov (United States)

    Colella, Rita; Hollensead, Sandra C

    2012-03-01

    The Pelger-Huët anomaly (PHA) is a recognized morphologic variant affecting all granulocytes but is most evident in polymorphonuclear neutrophils (PMNs). PHA is caused by a decreased amount of the lamin B receptor (LBR). Recognition of PHA morphologic features serves as a marker for mutations in the LBR gene. This review summarizes the history of PHA and the current knowledge of the functions of the LBR. Guidance is given for distinguishing PHA from other hematologic disorders in which granulocytes may show similar changes. Recognition of PHA in the laboratory should prompt communication to the patient's physician about the possible clinical significance of this finding and the recommended screening for the anomaly in other family members by CBC and review of a peripheral blood smear.

  12. Women for women's health: Uganda.

    Science.gov (United States)

    Andrews, C M

    1996-01-01

    The primary health care model targets social, political, and economic environments as key determinants of health for populations, as well as for individuals. If nursing in Uganda is to make a difference in health care outcomes and in the health of all Ugandans, nurses must look broadly at situations and be educated to practice primary health care nursing. After 14 years of civil war, Uganda is finally experiencing a period of reconstruction and rehabilitation: the whole infrastructure is undergoing a face-lift. Ugandan nurses recognize that their educational preparation has stagnated for many years and that it was not only the political unrest in their country that put them behind professionally. They realize that, given the new directions set by the government, they must become prepared to implement primary health care. They are demanding a university education so they may take their place alongside other health care providers prepared at the university level. Some of the most convincing arguments for a university program for nurses came from doctors at the university who spoke about the need to raise the standards of nursing practice, the quality of teachers, and the morale of practitioners. One nurse said: "If we lose hope for a BScN program, I think all the nurses will quit and we won't have any new students going into the profession." This program is designed to improve the health and well-being of all Ugandans, especially the most vulnerable groups of women and children in rural areas, through strengthening and expanding health services by targeting the educational preparation of nurses. Health planners in Uganda envision the professional nurse as key to the implementation of the national health policy of primary health care. University-educated nurses should be able to assess problems, make clinically sound decisions, and act appropriately within the scope of nursing practice. They should be able to interact and consult collegially with other health care

  13. Depression: women-at-risk.

    Science.gov (United States)

    Wetzel, J W

    1994-01-01

    Recognizing that women are three times more likely to experience depression than are men, this paper analyzes a spectrum of leading theories of depression through a feminist lens. Commonalities across perspectives are highlighted throughout, culminating in a synthesis. Facts, assumptions and issues regarding women and their high risk status are addressed in relation to each theory, providing insight concerning the many causes for their vulnerability, as well as assessing the ways in which each perspective sets up barriers to their development and well-being. By so doing, this prevention-oriented critique brings abstract ideas about depression into the realm of women's shared reality.

  14. Plasmodium falciparum: VAR2CSA expressed during pregnancy-associated malaria is partially resistant to proteolytic cleavage by trypsin

    DEFF Research Database (Denmark)

    Nielsen, Morten A; Resende, Mafalda; Alifrangis, Michael

    2007-01-01

    In areas of high Plasmodium falciparum transmission, immunity to malaria is acquired during childhood, so that adults in general are clinically immune. One exception is that first-time pregnant women are susceptible to pregnancy-associated malaria caused by accumulation of parasites in the placenta...... mediates the adhesive and antigenic phenotypes shown by parasites causing placental malaria....

  15. Recognizing disordered eating in primary care patients with obesity.

    Science.gov (United States)

    Chacko, Sara A; Chiodi, Sarah N; Wee, Christina C

    2015-03-01

    In clinical practice, behavioral approaches to obesity treatment focus heavily on diet and exercise recommendations. However, these approaches may not be effective for patients with disordered eating behaviors. Little is known about the prevalence of disordered eating behaviors in primary care patients with obesity or whether they affect difficulty making dietary changes. We conducted a telephone interview of 337 primary care patients aged 18-65 years with BMI ≥ 35 kg/m(2) in Greater Boston, 2009-2011 (58% response rate, 69% women). We administered the Three-Factor Eating Questionnaire R-18 (scores 0-100) and the Impact of Weight on Quality of Life-Lite (IWQOL-Lite) (scores 0-100). We measured difficulty making dietary changes using four questions regarding perceived difficulty changing diet (Scores 0-10). 50% of the patients reported high emotional eating (score>50) and 28% reported high uncontrolled eating (score>50). Women were more likely to report emotional [OR=4.14 (2.90, 5.92)] and uncontrolled eating [OR=2.11 (1.44, 3.08)] than men. African-Americans were less likely than Caucasians to report emotional [OR=0.29 (95% CI: 0.19, 0.44)] and uncontrolled eating [OR=0.11 (0.07, 0.19)]. For every 10-point reduction in QOL score (IWQOL-lite), emotional and uncontrolled eating scores rose significantly by 7.82 and 5.48, respectively. Furthermore, participants who reported emotional and uncontrolled eating reported greater difficulty making dietary changes. Disordered eating behaviors are prevalent among obese primary care patients and disproportionately affect women, Caucasians, and patients with poor QOL. These eating behaviors may impair patients' ability to make clinically recommended dietary changes. Clinicians should consider screening for disordered eating behaviors and tailoring obesity treatment accordingly. Copyright © 2015. Published by Elsevier Inc.

  16. Recognizing and treating delayed or failed lactogenesis II.

    Science.gov (United States)

    Hurst, Nancy M

    2007-01-01

    Delayed or failed achievement of lactogenesis II--the onset of copious milk volume--occurs as a result of various maternal and/or infant factors. Early recognition of these risk factors is critical for clinicians who interact with breastfeeding women so that intervention and achievement of full or partial breastfeeding can be preserved. This article describes the maternal and infant conditions that contribute to the unsuccessful establishment of a full lactation. Treatment modalities that can maximize maternal lactation capacity and infant growth rates are offered.

  17. Perspectives of digestive pest control with proteinase inhibitors that mainly affect the trypsin-like activity of Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae

    Directory of Open Access Journals (Sweden)

    M.E. Pereira

    2005-11-01

    Full Text Available The present study describes the main characteristics of the proteolytic activities of the velvetbean caterpillar, Anticarsia gemmatalis Hübner, and their sensitivity to proteinase inhibitors and activators. Midguts of last instar larvae reared on an artificial diet were homogenized in 0.15 M NaCl and centrifuged at 14,000 g for 10 min at 4ºC and the supernatants were used in enzymatic assays at 30ºC, pH 10.0. Basal total proteolytic activity (azocasein hydrolysis was 1.14 ± 0.15 absorbance variation min-1 mg protein-1, at 420 nm; basal trypsin-like activity (N-benzoyl-L-arginine-p-nitroanilide, BApNA, hydrolysis was 0.217 ± 0.02 mmol p-nitroaniline min-1 mg protein-1. The maximum proteolytic activities were observed at pH 10.5 using azocasein and at pH 10.0 using BApNA, this pH being identical to the midgut pH of 10.0. The maximum trypsin-like activity occurred at 50ºC, a temperature that reduces enzyme stability to 80 and 60% of the original, when pre-incubated for 5 and 30 min, respectively. Phenylmethylsulfonyl fluoride inhibited the proteolytic activities with an IC50 of 0.39 mM for azocasein hydrolysis and of 1.35 mM for BApNA hydrolysis. Benzamidine inhibited the hydrolysis with an IC50 of 0.69 and 0.076 mM for azocasein and BApNA, respectively. The absence of cysteine-proteinases is indicated by the fact that 2-mercaptoethanol and L-cysteine did not increase the rate of azocasein hydrolysis. These results demonstrate the presence of serine-proteinases and the predominance of trypsin-like activity in the midgut of Lepidoptera insects, now also detected in A. gemmatalis, and suggest this enzyme as a major target for pest control based on disruption of protein metabolism using proteinase inhibitors.

  18. No more violence against women.

    Science.gov (United States)

    1998-01-01

    Violence against women is now so widely recognized as a major obstacle to women's efforts to achieve "equality, development, and peace" that the UN General Assembly has decided to place the issue on its agenda this year. Viet Nam was one of the first signers of the 1982 Convention on the Elimination of All Forms of Discrimination Against Women, and the government has created a national committee for the advancement of women. This committee has established a national network to monitor implementation of the Convention. Among other government policies that are favorable to women is a national strategy for the advancement of women to the year 2000 that has 10 objectives including the elimination of violence against women and of all forms of discrimination against women. It is understood, however, that it is the responsibility of the entire society rather than of the government alone to ensure the rights of women and children. Currently, the incidence of child abuse and sex abuse is increasing in Viet Nam, especially in rural and remote areas. In response, the government is preparing to amend the penal code, and social organizations are campaigning to end violence. This combined approach to prevention seeks to raise social awareness about the equal roles played by women and men.

  19. Women's organizations: An untapped resource for resilience | CRDI ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    One South African organization explained, “Social crises like rampant violence against women, HIV/AIDS, access to land… are not looked at in the same way as natural crises. The urgency ... Programs also need to recognize women's care responsibilities and use solutions that do not overburden women. The research ...

  20. Feature Engineering for Recognizing Adverse Drug Reactions from Twitter Posts

    Directory of Open Access Journals (Sweden)

    Hong-Jie Dai

    2016-05-01

    Full Text Available Social media platforms are emerging digital communication channels that provide an easy way for common people to share their health and medication experiences online. With more people discussing their health information online publicly, social media platforms present a rich source of information for exploring adverse drug reactions (ADRs. ADRs are major public health problems that result in deaths and hospitalizations of millions of people. Unfortunately, not all ADRs are identified before a drug is made available in the market. In this study, an ADR event monitoring system is developed which can recognize ADR mentions from a tweet and classify its assertion. We explored several entity recognition features, feature conjunctions, and feature selection and analyzed their characteristics and impacts on the recognition of ADRs, which have never been studied previously. The results demonstrate that the entity recognition performance for ADR can achieve an F-score of 0.562 on the PSB Social Media Mining shared task dataset, which outperforms the partial-matching-based method by 0.122. After feature selection, the F-score can be further improved by 0.026. This novel technique of text mining utilizing shared online social media data will open an array of opportunities for researchers to explore various health related issues.

  1. Recognizing sights, smells, and sounds with gnostic fields.

    Directory of Open Access Journals (Sweden)

    Christopher Kanan

    Full Text Available Mammals rely on vision, audition, and olfaction to remotely sense stimuli in their environment. Determining how the mammalian brain uses this sensory information to recognize objects has been one of the major goals of psychology and neuroscience. Likewise, researchers in computer vision, machine audition, and machine olfaction have endeavored to discover good algorithms for stimulus classification. Almost 50 years ago, the neuroscientist Jerzy Konorski proposed a theoretical model in his final monograph in which competing sets of "gnostic" neurons sitting atop sensory processing hierarchies enabled stimuli to be robustly categorized, despite variations in their presentation. Much of what Konorski hypothesized has been remarkably accurate, and neurons with gnostic-like properties have been discovered in visual, aural, and olfactory brain regions. Surprisingly, there have not been any attempts to directly transform his theoretical model into a computational one. Here, I describe the first computational implementation of Konorski's theory. The model is not domain specific, and it surpasses the best machine learning algorithms on challenging image, music, and olfactory classification tasks, while also being simpler. My results suggest that criticisms of exemplar-based models of object recognition as being computationally intractable due to limited neural resources are unfounded.

  2. Periodontology as a recognized dental speciality in Europe.

    Science.gov (United States)

    Sanz, Mariano; van der Velden, Ubele; van Steenberghe, Daniel; Baehni, Pierre

    2006-06-01

    The impetus of the Bologna Process under the auspices of European Union governments has raised enormous expectations. It is the major educational change in Europe within the last 50 years and all the focus from university institutions, learned societies and thematic networks has shifted to this process, with the aim of developing consensus schemes in order to arrive at the expected European Convergence in Higher Education (to be completed by 2010). Dentistry as one of the health professions with clear Educational Standards, as defined by the European Dental Directives, is also reviewing its educational processes within this Bachelor-Master-Doctorate scheme and evaluating how the current and future dental specialities should be accommodated within this framework. Among these specialities, Periodontology is currently considered a formal dental speciality in 11 countries belonging to the EU however it lacks this legal status in the rest of the 14 EU countries. The purpose of this position paper is to provide evidence for the need for a recognized specialty in Periodontology at European level focusing on both the educational and professional perspective, with the hope of providing discussions that may contribute to facilitate its legal establishment as a new dental speciality in Europe.

  3. Random Deep Belief Networks for Recognizing Emotions from Speech Signals

    Directory of Open Access Journals (Sweden)

    Guihua Wen

    2017-01-01

    Full Text Available Now the human emotions can be recognized from speech signals using machine learning methods; however, they are challenged by the lower recognition accuracies in real applications due to lack of the rich representation ability. Deep belief networks (DBN can automatically discover the multiple levels of representations in speech signals. To make full of its advantages, this paper presents an ensemble of random deep belief networks (RDBN method for speech emotion recognition. It firstly extracts the low level features of the input speech signal and then applies them to construct lots of random subspaces. Each random subspace is then provided for DBN to yield the higher level features as the input of the classifier to output an emotion label. All outputted emotion labels are then fused through the majority voting to decide the final emotion label for the input speech signal. The conducted experimental results on benchmark speech emotion databases show that RDBN has better accuracy than the compared methods for speech emotion recognition.

  4. Recognizing emotions from EEG subbands using wavelet analysis.

    Science.gov (United States)

    Candra, Henry; Yuwono, Mitchell; Handojoseno, Ardi; Chai, Rifai; Su, Steven; Nguyen, Hung T

    2015-01-01

    Objectively recognizing emotions is a particularly important task to ensure that patients with emotional symptoms are given the appropriate treatments. The aim of this study was to develop an emotion recognition system using Electroencephalogram (EEG) signals to identify four emotions including happy, sad, angry, and relaxed. We approached this objective by firstly investigating the relevant EEG frequency band followed by deciding the appropriate feature extraction method. Two features were considered namely: 1. Wavelet Energy, and 2. Wavelet Entropy. EEG Channels reduction was then implemented to reduce the complexity of the features. The ground truth emotional states of each subject were inferred using Russel's circumplex model of emotion, that is, by mapping the subjectively reported degrees of valence (pleasure) and arousal to the appropriate emotions - for example, an emotion with high valence and high arousal is equivalent to a `happy' emotional state, while low valence and low arousal is equivalent to a `sad' emotional state. The Support Vector Machine (SVM) classifier was then used for mapping each feature vector into corresponding discrete emotions. The results presented in this study indicated thatWavelet features extracted from alpha, beta and gamma bands seem to provide the necessary information for describing the aforementioned emotions. Using the DEAP (Dataset for Emotion Analysis using electroencephalogram, Physiological and Video Signals), our proposed method achieved an average sensitivity and specificity of 77.4% ± 14.1% and 69.1% ± 12.8%, respectively.

  5. Diversity in warning coloration is easily recognized by avian predators.

    Science.gov (United States)

    Arenas, L M; Stevens, M

    2017-07-01

    Warning coloration is a widespread strategy to alert predators about prey unprofitability. The success of this strategy partly depends on predators being able to learn and recognize certain signals as indicators of toxicity, and theory predicts that this is easier if signals converge on similar colours. However, the diversity in warning signal form is astonishing, contradicting predictions. Here, we quantified ladybird signal diversity with respect to avian vision, measuring how unique and discernible each signal is from one another. In addition, we measured signal conspicuousness against a series of backgrounds, namely an average green, average brown, and where we collected each species, to determine whether signals are more contrasting against the ladybirds' local substrates than compared to average ones. This allowed us to establish whether there are local adaptations in conspicuousness that promote signal diversity. We found that while ladybird signals are unique and recognizable, specialist species are more contrasting against the background they are most commonly found on. However, overall our study suggests that warning signals have evolved to be effective against a wide range of natural backgrounds, partly explaining the success of this strategy in nature. © 2017 The Authors. Journal of Evolutionary Biology published by John Wiley & Sons Ltd on behalf of European Society for Evolutionary Biology.

  6. Simulation and psychology: creating, recognizing and using learning opportunities.

    Science.gov (United States)

    Dieckmann, Peter; Krage, Ralf

    2013-12-01

    Psychology is relevant for improving the use of simulation in anesthesiology, as it allows us to describe, explain and optimize the interactions of learners and instructors as well as the design of simulation scenarios and debriefings. Much psychological expertise is not used for simulation in healthcare. This article aims to help bridging the gap between professions. The evidence is building that simulation is effective for learning. Recent psychological work improves the understanding of why this is the case - or why not. Publications range from the elements to be simulated, to optimizing the presentation of scenarios and debriefings to an organizational overview of how simulation can contribute to patient safety, healthcare worker well-beings and quality of care. The psychological analysis helps in capturing the salient characteristics of the tasks to be simulated and in implementing them in a relevant learning setting. Using psychology in simulation allows us to create, recognize and use learning opportunities. The motivations of those involved can be taken into account and the simulation activity can be channeled into a goal-oriented direction. See the Video Supplementary Digital Content 1 (http://links.lww.com/COAN/A26).

  7. A deep convolutional neural network for recognizing foods

    Science.gov (United States)

    Jahani Heravi, Elnaz; Habibi Aghdam, Hamed; Puig, Domenec

    2015-12-01

    Controlling the food intake is an efficient way that each person can undertake to tackle the obesity problem in countries worldwide. This is achievable by developing a smartphone application that is able to recognize foods and compute their calories. State-of-art methods are chiefly based on hand-crafted feature extraction methods such as HOG and Gabor. Recent advances in large-scale object recognition datasets such as ImageNet have revealed that deep Convolutional Neural Networks (CNN) possess more representation power than the hand-crafted features. The main challenge with CNNs is to find the appropriate architecture for each problem. In this paper, we propose a deep CNN which consists of 769; 988 parameters. Our experiments show that the proposed CNN outperforms the state-of-art methods and improves the best result of traditional methods 17%. Moreover, using an ensemble of two CNNs that have been trained two different times, we are able to improve the classification performance 21:5%.

  8. Structural basis of Zika virus helicase in recognizing its substrates

    Directory of Open Access Journals (Sweden)

    Hongliang Tian

    2016-07-01

    Full Text Available Abstract The recent explosive outbreak of Zika virus (ZIKV infection has been reported in South and Central America and the Caribbean. Neonatal microcephaly associated with ZIKV infection has already caused a public health emergency of international concern. No specific vaccines or drugs are currently available to treat ZIKV infection. The ZIKV helicase, which plays a pivotal role in viral RNA replication, is an attractive target for therapy. We determined the crystal structures of ZIKV helicase-ATP-Mn2+ and ZIKV helicase-RNA. This is the first structure of any flavivirus helicase bound to ATP. Comparisons with related flavivirus helicases have shown that although the critical P-loop in the active site has variable conformations among different species, it adopts an identical mode to recognize ATP/Mn2+. The structure of ZIKV helicase-RNA has revealed that upon RNA binding, rotations of the motor domains can cause significant conformational changes. Strikingly, although ZIKV and dengue virus (DENV apo-helicases share conserved residues for RNA binding, their different manners of motor domain rotations result in distinct individual modes for RNA recognition. It suggests that flavivirus helicases could have evolved a conserved engine to convert chemical energy from nucleoside triphosphate to mechanical energy for RNA unwinding, but different motor domain rotations result in variable RNA recognition modes to adapt to individual viral replication.

  9. Recognizing genes and other components of genomic structure

    Energy Technology Data Exchange (ETDEWEB)

    Burks, C. (Los Alamos National Lab., NM (USA)); Myers, E. (Arizona Univ., Tucson, AZ (USA). Dept. of Computer Science); Stormo, G.D. (Colorado Univ., Boulder, CO (USA). Dept. of Molecular, Cellular and Developmental Biology)

    1991-01-01

    The Aspen Center for Physics (ACP) sponsored a three-week workshop, with 26 scientists participating, from 28 May to 15 June, 1990. The workshop, entitled Recognizing Genes and Other Components of Genomic Structure, focussed on discussion of current needs and future strategies for developing the ability to identify and predict the presence of complex functional units on sequenced, but otherwise uncharacterized, genomic DNA. We addressed the need for computationally-based, automatic tools for synthesizing available data about individual consensus sequences and local compositional patterns into the composite objects (e.g., genes) that are -- as composite entities -- the true object of interest when scanning DNA sequences. The workshop was structured to promote sustained informal contact and exchange of expertise between molecular biologists, computer scientists, and mathematicians. No participant stayed for less than one week, and most attended for two or three weeks. Computers, software, and databases were available for use as electronic blackboards'' and as the basis for collaborative exploration of ideas being discussed and developed at the workshop. 23 refs., 2 tabs.

  10. The repertoire of glycosphingolipids recognized by Vibrio cholerae.

    Directory of Open Access Journals (Sweden)

    John Benktander

    Full Text Available The binding of cholera toxin to the ganglioside GM1 as the initial step in the process leading to diarrhea is nowadays textbook knowledge. In contrast, the knowledge about the mechanisms for attachment of Vibrio cholerae bacterial cells to the intestinal epithelium is limited. In order to clarify this issue, a large number of glycosphingolipid mixtures were screened for binding of El Tor V. cholerae. Several specific interactions with minor complex non-acid glycosphingolipids were thereby detected. After isolation of binding-active glycosphingolipids, characterization by mass spectrometry and proton NMR, and comparative binding studies, three distinct glycosphingolipid binding patterns were defined. Firstly, V. cholerae bound to complex lacto/neolacto glycosphingolipids with the GlcNAcβ3Galβ4GlcNAc sequence as the minimal binding epitope. Secondly, glycosphingolipids with a terminal Galα3Galα3Gal moiety were recognized, and the third specificity was the binding to lactosylceramide and related compounds. V. cholerae binding to lacto/neolacto glycosphingolipids, and to the other classes of binding-active compounds, remained after deletion of the chitin binding protein GbpA. Thus, the binding of V. cholerae to chitin and to lacto/neolacto containing glycosphingolipids represents two separate binding specificities.

  11. Difficulty in recognizing multiple sulfatase deficiency in an infant.

    Science.gov (United States)

    Santos, Roberto P; Hoo, Joe J

    2006-03-01

    We describe the difficulty in recognizing multiple sulfatase deficiency (MSD; Online Mendelian Inheritance in Man [OMIM] database No. 272200) in an infant. MSD is a rare autosomal recessive disorder that affects the posttranslational activation of various sulfatase enzymes. It is both biochemically and clinically variable. Currently, there are 12 known sulfatases in humans, and the clinical presentation of MSD is a unique composite of those individual enzyme defects. Here we report a black girl who presented with bilateral broad thumbs and great toes, both with angulation deformities at birth. Rubinstein-Taybi syndrome (OMIM No. 180849) was considered initially. The detection of inclusion bodies in her white blood cells at 37 months of age led to the appropriate diagnostic workups for lysosomal storage diseases. Elevation of urine mucopolysaccharides provided additional clues, and the fibroblast enzyme assays finally established the diagnosis. Broad thumbs and great toes are rare features of MSD, and to the best of our knowledge such a bilateral congenital anomaly with angulation deformities has never been reported before to be associated with MSD.

  12. [The God of women].

    Science.gov (United States)

    Perez Aguirre, L

    1991-01-01

    The discourse of Christian theologians is by men and for men. The story of Salvation is about men; women have been excluded or colonized in ways carefully delimited by the Patriarchy. But a new struggle for liberation of women is underway worldwide in the dawn of the 21st century. The totality of relations between men and women is in change. The recent conquest of fertility control by women, which transferred ancestral male powers to them, and the decline of the Patriarchy are substantially modifying control of territory previously under male authority. The 2 revolutions are slowly but inexorably changing the social landscape. The feminization of poverty is of interest in this context. Women, 52% of the population, sow over half of food corps, account for 35% of the paid labor force and 60% of the hours worked, but receive only 10% of the income and possess only 1% of the world's property. Changes are occurring in the compulsively masculine culture, as well. A new consciousness and new intuitive knowledge of reality and its multiple cycles of change is emerging in which the individual is more satisfactorily related to the totality of the cosmos. Women in increasing numbers are freeing themselves of the known ways of the past which exalted the rational and mental to a new way which acknowledges the physical. In theology, the gods are beginning to appear as they were: true projections of the societies and structures of their times. Male theologians who reflect on these gods are always patriarchal because the church separates them from the world of women. They have been incapable as yet of comprehending the struggle of women who are opposing the world's oldest colonialism. For the same reasons they have been incapable of recognizing or accepting a fuller reality of existence and the divine. The prophetic female voice is still scarcely audible in an ecclesiastical world that remains enclosed in patriarch.

  13. Effect of gamma irradiation on lipoxygenases, trypsin inhibitor, raffinose family oligosaccharides and nutritional factors of different seed coat colored soybean (Glycine max L.)

    Energy Technology Data Exchange (ETDEWEB)

    Kumar Dixit, Amit [School of Biochemistry, Devi Ahilya University, Khandwa Road, Indore-452017 (India); Kumar, Vineet; Rani, Anita [National Research Centre for Soybean, Khandwa Road, Indore-452017 (India); Manjaya, J.G. [Board of Research in Nuclear Sciences, Bhabha Atomic Research Centre, Trombay, Mumbai (India); Bhatnagar, Deepak, E-mail: bhatnagarbio@yahoo.co [School of Biochemistry, Devi Ahilya University, Khandwa Road, Indore-452017 (India)

    2011-04-15

    Three soybean genotypes Kalitur, Hara soya and NRC37 with black, green and yellow seed coat color, respectively, were gamma irradiated at 0.5, 2.0 and 5.0 kGy and tested for antinutritional and nutritional factors. Gamma irradiation at all doses reduced the level of lipoxygenase isomers, trypsin inhibitor (TI) and ascorbic acid in all the 3 soybean genotypes as compared to the unirradiated control. However, irradiation dose of 5.0 kGy increased the sucrose content of the soybean genotypes. No significant change was observed in oil, protein fatty acids and total tocopherol content of the 3 genotypes at any irradiation dose. It is suggested that inhibition of lipoxygenase, reduction in TI and ascorbic acid may be due to the breakage or oxidation of protein structure by the gamma irradiation. Similarly, gamma irradiation at higher doses may break glycosidic linkages in oligosaccharides to produce more sucrose and decrease the content of flatulence causing oligosaccharides.

  14. Recognizing names in biomedical texts: a machine learning approach.

    Science.gov (United States)

    Zhou, GuoDong; Zhang, Jie; Su, Jian; Shen, Dan; Tan, ChewLim

    2004-05-01

    With an overwhelming amount of textual information in molecular biology and biomedicine, there is a need for effective and efficient literature mining and knowledge discovery that can help biologists to gather and make use of the knowledge encoded in text documents. In order to make organized and structured information available, automatically recognizing biomedical entity names becomes critical and is important for information retrieval, information extraction and automated knowledge acquisition. In this paper, we present a named entity recognition system in the biomedical domain, called PowerBioNE. In order to deal with the special phenomena of naming conventions in the biomedical domain, we propose various evidential features: (1) word formation pattern; (2) morphological pattern, such as prefix and suffix; (3) part-of-speech; (4) head noun trigger; (5) special verb trigger and (6) name alias feature. All the features are integrated effectively and efficiently through a hidden Markov model (HMM) and a HMM-based named entity recognizer. In addition, a k-Nearest Neighbor (k-NN) algorithm is proposed to resolve the data sparseness problem in our system. Finally, we present a pattern-based post-processing to automatically extract rules from the training data to deal with the cascaded entity name phenomenon. From our best knowledge, PowerBioNE is the first system which deals with the cascaded entity name phenomenon. Evaluation shows that our system achieves the F-measure of 66.6 and 62.2 on the 23 classes of GENIA V3.0 and V1.1, respectively. In particular, our system achieves the F-measure of 75.8 on the "protein" class of GENIA V3.0. For comparison, our system outperforms the best published result by 7.8 on GENIA V1.1, without help of any dictionaries. It also shows that our HMM and the k-NN algorithm outperform other models, such as back-off HMM, linear interpolated HMM, support vector machines, C4.5, C4.5 rules and RIPPER, by effectively capturing the local

  15. Evaluating the role of a trypsin inhibitor from soap nut (Sapindus trifoliatus L. Var. Emarginatus) seeds against larval gut proteases, its purification and characterization.

    Science.gov (United States)

    Gandreddi, V D Sirisha; Kappala, Vijaya Rachel; Zaveri, Kunal; Patnala, Kiranmayi

    2015-10-22

    The defensive capacities of plant protease Inhibitors (PI) rely on inhibition of proteases in insect guts or those secreted by microorganisms; and also prevent uncontrolled proteolysis and offer protection against proteolytic enzymes of pathogens. An array of chromatographic techniques were employed for purification, homogeneity was assessed by electrophoresis. Specificity, Ki value, nature of inhibition, complex formation was carried out by standard protocols. Action of SNTI on insect gut proteases was computationally evaluated by modeling the proteins by threading and docking studies by piper using Schrodinger tools. We have isolated and purified Soap Nut Trypsin Inhibitor (SNTI) by acetone fractionation, ammonium sulphate precipitation, ion exchange and gel permeation chromatography. The purified inhibitor was homogeneous by both gel filtration and polyacrylamide gel electrophoresis (PAGE). SNTI exhibited a molecular weight of 29 kDa on SDS-PAGE, gel filtration and was negative to Periodic Acid Schiff's stain. SNTI inhibited trypsin and pronase of serine class. SNTI demonstrated non-competitive inhibition with a Ki value of 0.75 ± 0.05×10-10 M. The monoheaded inhibitor formed a stable complex in 1:1 molar ratio. Action of SNTI was computationally evaluated on larval gut proteases from Helicoverpa armigera and Spodoptera frugiperda. SNTI and larval gut proteases were modeled and docked using Schrodinger software. Docking studies revealed strong hydrogen bond interactions between Lys10 and Pro71, Lys299 and Met80 and Van Der Waals interactions between Leu11 and Cys76amino acid residues of SNTI and protease from H. Armigera. Strong hydrogen bonds were observed between SNTI and protease of S. frugiperda at positions Thr79 and Arg80, Asp90 and Gly73, Asp2 and Gly160 respectively. We conclude that SNTI potentially inhibits larval gut proteases of insects and the kinetics exhibited by the protease inhibitor further substantiates its efficacy against serine

  16. A trypsin inhibitor from Tecoma stans leaves inhibits growth and promotes ATP depletion and lipid peroxidation in Candida albicans and Candida krusei

    Directory of Open Access Journals (Sweden)

    Leydianne Leite de Siqueira Patriota

    2016-04-01

    Full Text Available Tecoma stans (yellow elder has shown medicinal properties and antimicrobial activity. Previous reports on antifungal activity of T. stans preparations and presence of trypsin inhibitor activity from T. stans leaves stimulated the investigation reported here. In this work, we proceeded to the purification and characterization of a trypsin inhibitor (TesTI, which was investigated for anti-Candida activity. Finally, in order to determine the potential of TesTI as a new natural chemotherapeutic product, its cytotoxicity to human peripheral blood mononuclear cells (PBMCs was evaluated. TesTI was isolated from saline extract by ammonium sulphate fractionation followed by ion exchange and gel filtration chromatographies. Antifungal activity was evaluated by determining the minimal inhibitory (MIC and fungicide (MFC concentrations using fungal cultures containing only yeast form or both yeast and hyphal forms. Candida cells treated with TesTI were evaluated for intracellular ATP levels and lipid peroxidation. Cytotoxicity of TesTI to PBMCs was evaluated by MTT assay. TesTI (39.8 kDa, pI 3.41, Ki 43 nM inhibited similarly the growth of both C. albicans and C. krusei culture types at MIC of 100 µg/mL. The MFCs were 200 µg/mL for C. albicans and C. krusei. Time-response curves revealed that TesTI (at MIC was more effective at inhibiting the replication of C. albicans cells. At MIC, TesTI promoted reduction of ATP levels and lipid peroxidation in the Candida cells, being not cytotoxic to PBMCs. In conclusion, TesTI is an antifungal agent against C. albicans and C. krusei, without toxicity to human cells.

  17. A Trypsin Inhibitor from Tecoma stans Leaves Inhibits Growth and Promotes ATP Depletion and Lipid Peroxidation in Candida albicans and Candida krusei.

    Science.gov (United States)

    Patriota, Leydianne L S; Procópio, Thamara F; de Souza, Maria F D; de Oliveira, Ana Patrícia S; Carvalho, Lidiane V N; Pitta, Maira G R; Rego, Moacyr J B M; Paiva, Patrícia M G; Pontual, Emmanuel V; Napoleão, Thiago H

    2016-01-01

    Tecoma stans (yellow elder) has shown medicinal properties and antimicrobial activity. Previous reports on antifungal activity of T. stans preparations and presence of trypsin inhibitor activity from T. stans leaves stimulated the investigation reported here. In this work, we proceeded to the purification and characterization of a trypsin inhibitor (TesTI), which was investigated for anti-Candida activity. Finally, in order to determine the potential of TesTI as a new natural chemotherapeutic product, its cytotoxicity to human peripheral blood mononuclear cells (PBMCs) was evaluated. TesTI was isolated from saline extract by ammonium sulfate fractionation followed by ion exchange and gel filtration chromatographies. Antifungal activity was evaluated by determining the minimal inhibitory (MIC) and fungicide (MFC) concentrations using fungal cultures containing only yeast form or both yeast and hyphal forms. Candida cells treated with TesTI were evaluated for intracellular ATP levels and lipid peroxidation. Cytotoxicity of TesTI to PBMCs was evaluated by MTT assay. TesTI (39.8 kDa, pI 3.41, K i 43 nM) inhibited similarly the growth of both C. albicans and C. krusei culture types at MIC of 100 μg/mL. The MFCs were 200 μg/mL for C. albicans and C. krusei. Time-response curves revealed that TesTI (at MIC) was more effective at inhibiting the replication of C. albicans cells. At MIC, TesTI promoted reduction of ATP levels and lipid peroxidation in the Candida cells, being not cytotoxic to PBMCs. In conclusion, TesTI is an antifungal agent against C. albicans and C. krusei, without toxicity to human cells.

  18. Trypsin functionalization and zirconia coating of mesoporous silica nanotubes for matrix-assisted laser desorption/ionization mass spectrometry analysis of phosphoprotein.

    Science.gov (United States)

    Zhang, Xiaoli; Wang, Fei; Xia, Yan

    2013-09-06

    Trypsin functionalized mesoporous silica nanotubes bioreactor (TEMSN) and zirconia layer coated mesoporous silica nanotubes (ZrO2-MSN) were developed to deal with the long in-solution digestion time of phosphoprotein and detection difficulty of phosphorylated peptides, respectively. Trypsin was immobilized on the mesoporous silica nanotubes via epoxy group and TEMSN were used as a bioreactor for digestion of α-casein within 3min. ZrO2-MSN were performed to enrich phosphopeptides selectively from in-solution digested peptide mixture of β-casein to demonstrate that ZrO2-MSN possessed remarkable selectivity for phosphorylated peptides even at 100/1 molar ratio of BSA/β-casein. The selective ability of ZrO2-MSN was also investigated in comparison to ZrO2 nanoparticles (ZrO2 NP). Moreover, phosphorylated peptides at the femtomole (2.5fmol) level can also be detected with high S/N (signal-to-noise) ratio. Phosphopeptides enriched from TEMSN-bioreactor digested peptide mixture of α-casein was also performed to evaluate the cooperative performance of TEMSN and ZrO2-MSN platform. The experimental results indicated that TEMSN-bioreactor digestion changed the distribution of relative abundance of phosphopeptides and improved the relative intensity of partial phosphopeptides. This analytical strategy has also been applied to the identification of phosphopeptides isolated from non-fat bovine milk and got a comparable results compared with other materials cited from the literature. By matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS), TEMSN and ZrO2-MSN were combined together for the rapid and comprehensive analysis of phosphoprotein. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. Aspects of abuse: recognizing and responding to child maltreatment.

    Science.gov (United States)

    Jackson, Allison M; Kissoon, Natalie; Greene, Christian

    2015-03-01

    Child maltreatment is a public health problem and toxic stress impacting at least 1 in 8 children by the age of 18 years. Maltreatment can take the form of physical and sexual abuse, neglect, and emotional maltreatment. While some children may experience only one form of maltreatment, others may survive multiple forms, and in some cases particularly complex forms of maltreatment such as torture and medical child abuse. When considering maltreatment, providers should be adept at obtaining a thorough history not only from the parent but when appropriate also from the patient. The most common form of child maltreatment is neglect, which encompasses nutritional and medical neglect, as well as other forms such as physical and emotional neglect. Talking with caregivers about stressors and barriers to care may give insight into the etiology for neglect and is an opportunity for the provider to offer or refer for needed assistance. Familiarity with injury patterns and distribution in the context of developmental milestones and injury mechanisms is critical to the recognition of physical abuse. While most anogenital exam results of child victims of sexual abuse are normal, knowing the normal variations for the female genitalia, and thereby recognizing abnormal findings, is important not only forensically but also more importantly for patient care. Pattern recognition does not only apply to specific injuries or constellation of injuries but also applies to patterns of behavior. Harmful patterns of behavior include psychological maltreatment and medical child abuse, both of which cause significant harm to patients. As health professionals serving children and families, pediatric providers are in a unique position to identify suspected maltreatment and intervene through the health care system in order to manage the physical and psychological consequences of maltreatment and to promote the safety and well-being of children and youth by making referrals to child protective

  20. Celiac Crisis: A Rare Or Rarely Recognized Disease.

    Science.gov (United States)

    Waheed, Nadia; Cheema, Huma Arshad; Suleman, Hassan; Fayyaz, Zafar; Mushtaq, Iqra; Muhammad; Hashmi, Almas

    2016-01-01

    Celiac crisis is a serious life threatening complication of celiac disease characterized by profuse diarrhoea, severe dehydration and metabolic disturbances leading to neuromuscular weakness, cardiac arrhythmias and sudden death. It has been described as rare condition and not well documented in the literature. To improve awareness and facilitate diagnosis of this condition, we studied risk factors, pattern of presentation and management plans of celiac crisis. It was a descriptive cross sectional study. Patients presenting in emergency room(ER) with profuse diarrhoea leading to severe dehydration, neuromuscular weakness, and metabolic acidosis and electrolyte abnormalities enrolled in the studies after positive serology and small bowel biopsy suggestive of celiac disease. Total 126 patients out of 350 fulfilled the criteria including 54 (42.8%) male and 71 (56.3%) female. The mean age at presentation was 5.25±1.18 years. Risk factors were poor social status (97.60%), consanguinity (96.77%), early weaning with gluten contained diet (93.54%), and Presenting complaints were loose motion (100%), loss of neck holding (96.77%), dehydration (96.77%), polyuria (95.96%), inability to walk (67.74%), abdominal distension (85.86%). Electrolytes imbalances were hypokalaemia (2.4±0.55), hypocalcaemia (7.29±0.66), hypomagnesaemia (1.89±0.50), hypophosphatemia (2.8±0.68), hypoalbuminemia (3.05±0.48) and metabolic acidosis (96%). One hundred & twenty patients were stabilized with GFD and correction of dehydration, acidosis and electrolyte imbalance. Six patients needed parenteral steroids ant total parenteral nutrition (TPN). Recovery time from crisis was mean 5.4±2.73 days (range 3-20 days). Celiac crisis is a common but under recognized problem in developing countries. Commonest presenting feature is neuromuscular paralysis and biochemical abnormality is hypokalaemia.

  1. Effect of condensed tannins in hulls of faba beans (Vivia faba L.) on the activities of trypsin (EC 2.4.21.4) and chymotrypsin (EC 2.4.21.1) in digesta collected from the small intestine of pigs

    NARCIS (Netherlands)

    Jansman, A.J.M.; Enting, H.; Verstegen, M.W.A.; Huisman, J.

    1994-01-01

    The effects of condensed tannins in hulls of faba beans (Vicia faba L.) on the activities of trypsin (EC 3.4.21.4) and chymotrypsin (EC 3.4.21.1) in digesta obtained from the small intestine of pigs were studied. Using four castrated male pigs (mean body weight 83 kg) fitted with both a simple

  2. [Some insufficiently recognized risk factors for breast cancer].

    Science.gov (United States)

    Kocić, B; Janković, S; Petrović, B; Tiodorović, B

    1996-01-01

    In the most developed countries female breast cancer is the main cause of death due to the most common malignancy in women [1]. Over the period 1975-1982 and 1982-1991 women in Serbia died most frequently of malignant tumours of the breast [2, 3]. A tendency to increase mortality rate of the disease was recorded in this period. The aim of this paper was to determine the importance of some insufficiently known breast cancer risk factors. The study was based on a case-control study conducted from July 1993 to March 1994. The patients were selected at the Department of Oncology in Nish, and included 106 female patients with histologically documented breast cancer (a two-year diagnosis). Controls were individually matched to patients by age (+/-3 years) and number of patients with mild injuries, hospitalized at the Clinical Centre in Nish. All the examined subjects, from both groups, were interviewed by the same doctor using a target and detailed questionnaire about different risk factors of breast cancer. McNemar test was used in the study. The results are presented in Table 1. The results confirmed a significant positive association between breast cancer risk and psychological stressogenic events (RR = 4.40 95% CI = 2.30-8.40 p = 0.0000). With reference to a previous history of mastitis, a frequent positive history was found in the examined patients and it was statistically significant (RR = 7.50 95% CI = 2.99-18.83 p = 0.000). The risk of developing breast cancer was strongly related to alcohol consumption (RR = 4.11 95% CI = 2.05-8.24 p = 0.0001). Two environmental factors were significantly associated with breast cancer: occupational exposure to organic dust (RR = 3.80 95% CI = 1.41-10.20 p = 0.0066) and to pesticides (RR = 4.25 95% CI = 1.43-12.58 p = 0.0072). In our patients stressful life-events were significantly frequent. Most of them reported death or a serious illness of the spouse or of a close family member, marital divorce, son's military service, etc

  3. Mycophenolate mofetil embryopathy: A newly recognized teratogenic syndrome.

    Science.gov (United States)

    Perez-Aytes, Antonio; Marin-Reina, Purificacion; Boso, Virginia; Ledo, Ana; Carey, John C; Vento, Maximo

    2017-01-01

    Mycophenolate mofetil (MMF) is probably the most common employed immunosuppressant drug in recipients of solid organ transplant and in many autoimmune diseases. In vitro studies, a significant number of single clinical observations and a recent study from a group of different European teratogen information services, have provided very consistent data supporting the existence of a specific MMF embryopathy. The typical malformative pattern of MMF embryopathy includes external ear anomalies ranging from hypoplastic pinna (microtia) to complete absence of pinna (anotia); cleft lip, with or without cleft palate, and ocular anomalies as iris or chorioretinal coloboma and anophthalmia/microphthalmia. Other less frequent features are congenital heart defects, distal limbs anomalies, esophageal atresia, vertebral malformations, diaphragmatic hernia, and kidney and central nervous system anomalies. Neurodevelopmental outcome seems favorable in the small number of patients where information about this issue is available, but neurological deficits have been documented. Physicians in charge of women under MMF therapy should be aware of the potential risk of this drug to cause a specific embryopathy and the need of interrupting the treatment at least six weeks before becoming pregnant. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  4. Newly recognized submarine slide complexes in the southern California Bight

    Science.gov (United States)

    Conrad, J. E.; Lee, H. J.; Edwards, B. D.; McGann, M.; Sliter, R. W.

    2012-12-01

    terrestrial sediment input from fluvial sources, as are most other previously recognized submarine slides in the Borderland. Only the SPS slide, which lies adjacent to the San Gabriel Canyon submarine channel, is associated with a significant fluvial sediment source.

  5. 78 FR 60898 - Regulation on Definition and Requirements for a Nationally Recognized Testing Laboratory...

    Science.gov (United States)

    2013-10-02

    ... Occupational Safety and Health Administration Regulation on Definition and Requirements for a Nationally Recognized Testing Laboratory; Revision of the Office of Management and Budget's (OMB) Approval of..., ``definition and requirements for a nationally recognized testing laboratory'' (The Regulation). The Regulation...

  6. Recognize and classify pneumoconiosis; Pneumokoniosen erkennen und klassifizieren

    Energy Technology Data Exchange (ETDEWEB)

    Hering, K.G.; Hofmann-Preiss, K. [Klinikum Westfalen, Knappschaftskrankenhaus, Dortmund (Germany)

    2014-12-15

    In the year 2012, out of the 10 most frequently recognized occupational diseases 6 were forms of pneumoconiosis. With respect to healthcare and economic aspects, silicosis and asbestos-associated diseases are of foremost importance. The latter are to be found everywhere and are not restricted to large industrial areas. Radiology has a central role in the diagnosis and evaluation of occupational lung disorders. In cases of known exposure mainly to asbestos and quartz, the diagnosis of pneumoconiosis, with few exceptions will be established primarily by the radiological findings. As these disorders are asymptomatic for a long time they are quite often detected as incidental findings in examinations for other reasons. Therefore, radiologists have to be familiar with the pattern of findings of the most frequent forms of pneumoconiosis and the differential diagnoses. For reasons of equal treatment of the insured a quality-based, standardized performance, documentation and evaluation of radiological examinations is required in preventive procedures and evaluations. Above all, a standardized low-dose protocol has to be used in computed tomography (CT) examinations, although individualized concerning the dose, in order to keep radiation exposure as low as possible for the patient. The International Labour Office (ILO) classification for the coding of chest X-rays and the international classification of occupational and environmental respiratory diseases (ICOERD) classification used since 2004 for CT examinations meet the requirements of the insured and the occupational insurance associations as a means of reproducible and comparable data for decision-making. (orig.) [German] Im Jahr 2012 waren 6 der 10 am haeufigsten anerkannten Berufskrankheiten Pneumokoniosen. Gesundheitspolitisch und oekonomisch stehen dabei die Silikose und asbestassoziierte Erkrankungen im Vordergrund. Insbesondere Letztere treten ubiquitaer auf und sind nicht an grosse Industriestandorte gebunden

  7. Sex Differences in Music: A Female Advantage at Recognizing Familiar Melodies.

    Science.gov (United States)

    Miles, Scott A; Miranda, Robbin A; Ullman, Michael T

    2016-01-01

    Although sex differences have been observed in various cognitive domains, there has been little work examining sex differences in the cognition of music. We tested the prediction that women would be better than men at recognizing familiar melodies, since memories of specific melodies are likely to be learned (at least in part) by declarative memory, which shows female advantages. Participants were 24 men and 24 women, with half musicians and half non-musicians in each group. The two groups were matched on age, education, and various measures of musical training. Participants were presented with well-known and novel melodies, and were asked to indicate their recognition of familiar melodies as rapidly as possible. The women were significantly faster than the men in responding, with a large effect size. The female advantage held across musicians and non-musicians, and across melodies with and without commonly associated lyrics, as evidenced by an absence of interactions between sex and these factors. Additionally, the results did not seem to be explained by sex differences in response biases, or in basic motor processes as tested in a control task. Though caution is warranted given that this is the first study to examine sex differences in familiar melody recognition, the results are consistent with the hypothesis motivating our prediction, namely that declarative memory underlies knowledge about music (particularly about familiar melodies), and that the female advantage at declarative memory may thus lead to female advantages in music cognition (particularly at familiar melody recognition). Additionally, the findings argue against the view that female advantages at tasks involving verbal (or verbalizable) material are due solely to a sex difference specific to the verbal domain. Further, the results may help explain previously reported cognitive commonalities between music and language: since declarative memory also underlies language, such commonalities may be

  8. Sex Differences in Music: A Female Advantage at Recognizing Familiar Melodies

    Directory of Open Access Journals (Sweden)

    Michael T Ullman

    2016-03-01

    Full Text Available Although sex differences have been observed in various cognitive domains, there has been little work examining sex differences in the cognition of music. We tested the prediction that women would be better than men at recognizing familiar melodies, since memories of specific melodies are likely to be learned (at least in part by declarative memory, which shows female advantages. Participants were 24 men and 24 women, with half musicians and half non-musicians in each group. The two groups were matched on age, education, and various measures of musical training. Participants were presented with well-known and novel melodies, and were asked to indicate their recognition of familiar melodies as rapidly as possible. The women were significantly faster than the men in responding, with a large effect size. The female advantage held across musicians and non-musicians, and across melodies with and without commonly associated lyrics, as evidenced by an absence of interactions between sex and these factors. Additionally, the results did not seem to be explained by sex differences in response biases, or in basic auditory or motor processes as tested in a control task. Though caution is warranted given that this is the first study to examine sex differences in familiar melody recognition, the results are consistent with the hypothesis motivating our prediction, namely that declarative memory underlies knowledge about music (particularly about familiar melodies, and that the female advantage at declarative memory may thus lead to female advantages in music cognition (particularly at familiar melody recognition. Additionally, the findings argue against the view that female advantages at tasks involving verbal (or verbalizable material are due solely to a sex difference specific to the verbal domain. Further, the results may help explain previously-reported cognitive commonalities between music and language: since declarative memory also underlies language

  9. 29 CFR 779.368 - Printing and engraving establishments not recognized as retail.

    Science.gov (United States)

    2010-07-01

    ... 29 Labor 3 2010-07-01 2010-07-01 false Printing and engraving establishments not recognized as... Service Establishments Commercial Stationers § 779.368 Printing and engraving establishments not recognized as retail. (a) An establishment which is engaged in printing and engraving is not recognized as a...

  10. Biographies of Eminent Women in Psychology: Models for Achievement.

    Science.gov (United States)

    Furumoto, Laurel; And Others

    1980-01-01

    In order to recognize women's contributions to the field of psychology, biographies of Margaret Floy Washburn, Mary Cover Jones, Karen Horney, Susan Grey, Edna Heidbreder, Ann Roe, and Mary Whitton Calkins are presented. (BEF)

  11. The Status of Women in Physics in the U.S.: Progress and New Actions

    Science.gov (United States)

    Xie, Aihua

    2005-10-01

    The 2005 Chair of the American Physical Society Committee on the Status of Women in Physics describes the committee's recent activities to recognize distinguished women physicists, improve the climate for women in physics, and provide leadership training for women in physics. The committee's response to the Harvard University president's suggestion of innate gender differences as regards women's representation in math and science is also discussed, as well as some encouraging developments in the status of women in physics in the U.S.

  12. Women?s perceptions and experiences of the challenges in the process of male infertility treatment: A qualitative study

    OpenAIRE

    Taghipour, Ali; Karimi, Fatemeh Zahra; Roudsari, Robab Latifnejad; Kimiaei, Seyed Ali; Mazlom, Seyed Reza; Amirian, Maliheh

    2017-01-01

    Background Despite the fact that both men and women are equally subject to infertility, it is usually women who bear the burden of treatment and its consequences, even in cases of male infertility. Therefore, it is more necessary to recognize their health problems in order to help them. Aim To explore women?s perceptions and experiences of the challenges they face in the process of male infertility treatment. Methods This qualitative study was conducted during 2014?2015 using content analysis...

  13. Women boxers

    DEFF Research Database (Denmark)

    Gems, Gerald; Pfister, Gertrud Ursula

    2014-01-01

    This article fills a gap in the very limited literature on women's boxing by examining the gendered space in which women engaged in the sport as participants in saloons, vaudeville theatres and the prize ring. In doing so, they challenged the contemporary gender order and disputed the notion...... of women as the weak sex. Vaudeville provided women with an opportunity to present physical performances that surpassed the restrictions placed on women within the mainstream middle-class society. This article includes biographical sketches of some of the outstanding female boxers of the era by drawing...

  14. International women's movement comes of age at FWCW.

    Science.gov (United States)

    O'haire, H

    1996-02-01

    This article focuses on what the women's conference achieved for women. The Fourth World Conference on Women (FWCW) recognized an international women's movement. Governments agreed that women were entitled to full human rights, reproductive rights, and equality in all aspects of life. Women were to become full and equal partners in the development process. Women are likely to demand action based on the paper declarations. The success of the conference is attributed to a powerful network of women's groups. A striking feature was the heading of most national delegations by women. It was recognized that women were full but neglected partners in families and societies. Governments acknowledged that women suffered discrimination within the family and society. Wording was rejected that would have given women the guaranteed right to determine the size and spacing of their families and the information and services to do so. The activity among women's groups in working together to restore this view strengthened the bonds between women's groups and created worldwide awareness and support for women's organizations. Networks that were established at Rio de Janeiro made their presence and position on reproductive health felt at the preparatory meetings to the UN Population Conference in Cairo. It was argued that a target-driven approach had the effect of treating women as reproduction machines. Women's groups also expressed strong positions on how women were to be treated in the formation of population policy and programs. 1) It was insisted that governments and population groups must stop dictating fertility regimes to women. 2) The concept of family planning must include reproductive health. 3) Women should have the freedom to exercise choice in planning their families. 4) Women should also have equal access to education and employment. Women came thus to Beijing with a clear vision of what they wanted. At Beijing domestic violence was for the first time condemned. Beijing

  15. Karakterisasi stem cell pulpa gigi sulung dengan modifikasi enzim tripsin (The characterization of stem cells from human exfoliated deciduous teeth using trypsin enzym

    Directory of Open Access Journals (Sweden)

    Tri Wijayanti Puspitasari

    2014-06-01

    Full Text Available Background: Now a days, treatment in dentistry, using tissue regeneration that based on the stem cells from human exfoliated deciduous teeth (SHED, grows rapidly. For several reason, the isolated and cultured SHED is difficult to be applied in Indonesia, therefore the modification is needed. This difficulties were caused by the pulp anatomy, the heterogeneous populations in the pulp chamber and the limitations of tools and materials at the laboratory. Purpose: This research was aimed to examine that the modifications of isolation and culture technique of SHEDs for characterization by using the marker of CD105. Methods: The research was experimental laboratory with the cross sectional design. The samples were the human exfoliated deciduous teeth from the children patients of Pediatric Dentistry Department of Universitas Airlangga Dental Hospital which matched the criteria. Dental pulps were isolated and cultured by using the modifications of Trypsin enzymes. Results: The healthy SHEDs could be produced from the modifications of isolation and culture and positively shown the expression of marker CD105 which were indicated by the fluorencent microscope. Conclusion: SHED which isolated and cultured by using the modified techniques, positively characterized by using marker CD105.Latar Belakang: Pengobatan kedokteran gigi berkembang dengan pesat terutama di bidang regenerasi jaringan berbasis Stem Cells from Human Exfoliated Deciduous Teeth (SHED. Di Indonesia, isolasi dan kultur SHED sulit sehingga perlu dilakukan modifikasi. Kendala ini muncul karena jaringan pulpa yang kecil, heterogen dan keterbatasan alat dan bahan di laboratorium. Tujuan: Penelitian ini bertujuan untuk meneliti modifikasi pada cara isolasi dan kultur SHED untuk karakterisasi menggunakan maker CD105. Metode: Jenis penelitian ini adalah eksperimental laboratoris dengan rancangan cross sectional. Sampel penelitian adalah gigi sulung dari pasien anak di Klinik Kedokteran Gigi Anak

  16. Von Willebrand disease and other bleeding disorders in women: consensus on diagnosis and management from an international expert panel

    NARCIS (Netherlands)

    James, Andra H.; Kouides, Peter A.; Abdul-Kadir, Rezan; Edlund, Mans; Federici, Augusto B.; Halimeh, Susan; Kamphuisen, Pieter W.; Konkle, Barbara A.; Martínez-Perez, Oscar; McLintock, Claire; Peyvandi, Flora; Winikoff, Rochelle

    2009-01-01

    Reproductive tract bleeding in women is a naturally occurring event during menstruation and childbirth. In women with menorrhagia, however, congenital bleeding disorders historically have been underdiagnosed. This consensus is intended to allow physicians to better recognize bleeding disorders as a

  17. Women, conflict, and culture in former Yugoslavia.

    Science.gov (United States)

    Stojsavljevic, J

    1995-02-01

    The civil war in the former Yugoslavia has taken a toll on the women's movement which has disintegrated across male-defined nationalist borders. The women's movement in this area got its start during the Second World War but was disbanded under communism until women's groups began to form in the 1970s. Today the women's movement has lost the power to oppose the war and has been unable to prevent widespread violence perpetuated against women. Some feminists who have refused to embrace nationalism and patriotism have been vilified and have had to seek refuge abroad. Recently, however, hundreds of nongovernmental organizations have been formed to provide support to women and children victimized by the war. Women have been raped and impregnated as a strategy of male warfare, and raped women who refused an abortion were ostracized. War-related rape has yet to be fully recognized as an international human rights violation, and the issue is being used as political propaganda in the former Yugoslavia while it is ignored elsewhere. Sensationalist reporting of these rapes has further victimized women and made them unable to give voice to their trauma. War also increases women's suffering by destroying economic and social welfare systems. Oxfam is helping women record their testimonies of war and reconstruct the fabric of their societies through programs which provide income-generation and training in micro-enterprises. In addition, Oxfam is strengthening electronic communication and networking among women's groups throughout the region.

  18. Does the oxidation of methionine residue precede the inactivation of the trypsin inhibitor (LUTI in germinating seeds of common flax (Linum usitatissimum?

    Directory of Open Access Journals (Sweden)

    Irena Lorenc-Kubis

    2011-01-01

    Full Text Available Antitrypsin activity in germinating common seeds of flax (Linum usitatissimum was investigated. At the early stage of germination an increase in antitrypsin activity was observed, followed by its decrease during the development of the seedlings. From 6-day-old seedlings a trypsin inhibitor (gerLUTI was purified. The purification procedure involved fractionation of proteins from seedling homogenate with alcohol and successive chromatography on CM-Sephadex C-25 on immobilised methylchymotrypsin in the presence of 5 M NaCl, and finally on a C18 column in RP-HPLC. The gerLUTI migrated in SDS PAGE as a single band, but in mass spectroscopy analysis it exhibited the presence of at least three forms with molecular masses of 7654 ± 3 Da, 7668/7670 ± 3 Da, and 7687 ± 3 Da. The preparation of LUTI isolated from resting seeds contained only one form, with a molecular mass of 7655 ± 3 Da. LUTI and gerLUTI differed also in methionine contents. LUTI contained two methionine residues, whereas in gerLUTI only a trace of methionine was detected. The obtained results might suggest that during flax seeds germination the inhibitor molecules undergo selective modification, e.g. oxidation at methionine residues, before being degraded by proteolytic enzymes.

  19. Interrogating and predicting tolerated sequence diversity in protein folds: application to E. elaterium trypsin inhibitor-II cystine-knot miniprotein.

    Directory of Open Access Journals (Sweden)

    Jennifer L Lahti

    2009-09-01

    Full Text Available Cystine-knot miniproteins (knottins are promising molecular scaffolds for protein engineering applications. Members of the knottin family have multiple loops capable of displaying conformationally constrained polypeptides for molecular recognition. While previous studies have illustrated the potential of engineering knottins with modified loop sequences, a thorough exploration into the tolerated loop lengths and sequence space of a knottin scaffold has not been performed. In this work, we used the Ecballium elaterium trypsin inhibitor II (EETI as a model member of the knottin family and constructed libraries of EETI loop-substituted variants with diversity in both amino acid sequence and loop length. Using yeast surface display, we isolated properly folded EETI loop-substituted clones and applied sequence analysis tools to assess the tolerated diversity of both amino acid sequence and loop length. In addition, we used covariance analysis to study the relationships between individual positions in the substituted loops, based on the expectation that correlated amino acid substitutions will occur between interacting residue pairs. We then used the results of our sequence and covariance analyses to successfully predict loop sequences that facilitated proper folding of the knottin when substituted into EETI loop 3. The sequence trends we observed in properly folded EETI loop-substituted clones will be useful for guiding future protein engineering efforts with this knottin scaffold. Furthermore, our findings demonstrate that the combination of directed evolution with sequence and covariance analyses can be a powerful tool for rational protein engineering.

  20. Effects of soybean trypsin inhibitor on hypopharyngeal gland protein content, total midgut protease activity and survival of the honey bee (Apis mellifera L.).

    Science.gov (United States)

    Sagili, Ramesh R; Pankiw, Tanya; Zhu-Salzman, Keyan

    2005-09-01

    Insecticidal properties of protease inhibitors have been established in transgenic plants. In the wake of continuous research and rapid development of protease inhibitors it is important to assess possible effects on beneficial insects like the honey bee (Apis mellifera L.). In this study, newly emerged caged bees were fed pollen diets containing three different concentrations (0.1%, 0.5% and 1% w:w) of soybean trypsin inhibitor (SBTI). Hypopharyngeal gland protein content, total midgut proteolytic enzyme activity of these bees, and survival were measured. Bees fed 1% SBTI had significantly reduced hypopharyngeal gland protein content and midgut proteolytic enzyme activity. There were no significant differences between control, 0.1% and 0.5% SBTI treatments. Bees fed a diet containing 1% SBTI had the lowest survival, followed by 0.5% and 0.1%, over a 30-day period. We concluded that nurse bees fed a pollen diet containing at least 1% SBTI would be poor producers of larval food, potentially threatening colony growth and maintenance.

  1. Mung bean trypsin inhibitor is effective in suppressing the degradation of myofibrillar proteins in the skeletal muscle of blue scad (Decapterus maruadsi).

    Science.gov (United States)

    Sun, Le-Chang; Yoshida, Asami; Cai, Qiu-Feng; Liu, Guang-Ming; Weng, Ling; Tachibana, Katsuyasu; Su, Wen-Jin; Cao, Min-Jie

    2010-12-22

    Mung bean trypsin inhibitor (MBTI) of the Bowman-Birk family was purified to homogeneity with a molecular mass of approximately 9 kDa on tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and 8887.25 Da as determined by matrix-assisted laser desorption/ionization-quadrupole ion trap-time-of-flight mass spectrometry (MALDI-QIT-TOF MS). Using blue scad myofibrillar proteins as targets, it was found that, in the absence of MBTI, proteolysis of myofibrillar proteins, especially myosin heavy chain (MHC), could be identified after incubation at 55 °C for 2 h, while in the presence of MBTI, with a final concentration of 25 ng/mL, proteolysis of these proteins was greatly suppressed even after incubation for 3 h. Although cysteine proteinase inhibitor E-64 was also effective in preventing protein degradation, inhibitors for metallo- and asparatic proteinases did not reveal obvious inhibitory effects. Our present results strongly suggested that the naturally occurring legume bean seed protein MBTI can be used as an effective additive in preventing marine fish blue scad surimi gel softening, which is quite possibly caused by myofibril-bound serine proteinase (MBSP).

  2. 46 CFR 159.010-7 - Recognized independent laboratory: Memorandum of Understanding.

    Science.gov (United States)

    2010-10-01

    ... Independent Laboratory: Acceptance, Recognition, and Termination § 159.010-7 Recognized independent laboratory... appropriate requirements, the Commandant or authorized representative may at his discretion sign the MOU...

  3. Women, population and development: rural women's passage to empowerment.

    Science.gov (United States)

    Villareal, F L

    1995-12-01

    This article describes the Women's Population and Development (WPD) program among rural women in 35 poor counties in 10 provinces in China in 1988. The provinces included Xinjiang, Gansu, Qinghai, Ningxia, Inner Mongolia, Shaanxi, Shanxi, Guizhou, Hubei, and Anhui, which are mostly located in the arid northwest or pastoral northeast China. The study area is inclined to have harsh agro-ecological conditions and environmental degradation. Women's income ranges from RMB 350 to 650 Yuan. About 35% were literate. 40-50% of the rural agricultural labor force and about 60% of all farm labor were women. Out migration of men was increasing. This WPD program was supported by the Ministry of Foreign Technical and Economic Cooperation, the UNFPA, and the UN Food and Agriculture Organization. The program aim is to use alternative family planning strategies, alleviate poverty, and improve rural women's status. Initial discussions revealed different goals among rural women and county leaders. Women wanted direct access and control over resources. County leaders desired increased productivity among a selected number of small scale enterprises. The project stipulated that 80% of funding was to be allocated as credit for rural women, that a revolving fund would give women direct access and control of resources, and that women would be recognized for their dual roles. Partnerships were formed between women's voluntary groups and project management. Prospective members of women's self-help groups received a 60-70 hour workshop. This workshop helped to identity the root causes of low status and feasible approaches to improving the status and to developed program activities. Counties were gradually included during 1990-93. This program evaluation pertains to analysis of 1994-95 data among 1300 respondents at four points in time. Impact indicators include 18 measures. The most important measure is that women gained control over their own lives and a partnership with husbands. The program

  4. Women's Rights and Living Customary Law | IDRC - International ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Women's Rights and Living Customary Law. This action-research project focuses on the interface between custom and rights in the context of a constitution that recognizes and protects both customary law and the Bill of Rights. It will explore how this interplay affects the rights - particularly land rights - of black women living ...

  5. Women in Student Affairs: Past, Present, and Future Leaders

    Science.gov (United States)

    McNair, Delores E.; Miguel, Krystal; Sobers-Young, Shauna T.; Bechtel, Molly; Jacobson, Steve

    2013-01-01

    In 2010, three women from the University of the Pacific came together for a panel presentation at the annual National Association of Student Personnel Administrators (NASPA) conference to discuss their diverse experiences in student affairs. All recognized leaders in NASPA, these women reflected the rich history and promising future of student…

  6. Balancing Acts for African Women Development: Challenges of ...

    African Journals Online (AJOL)

    In traditional African society as well as modern, women had, and still have, recognized roles in the socio- cultural as well as economic spheres. ... Since African female academics are empowered and inclined towards empowering other women, they need to remain healthy, improve their thinking and increase their energy ...

  7. equality idrc on for girls and women

    International Development Research Centre (IDRC) Digital Library (Canada)

    Cathy Egan

    In fact, many forms of violence, such as sexual and reproductive violence, forced. “marital” unions, and different forms of domestic enslavement, have not been adequately recognized as human rights violations — and have been left out of reparations programs. “Women (also) must own the reparations process, especially ...

  8. Nursing diagnoses in women deprived of freedom

    National Research Council Canada - National Science Library

    Izabelle de Freitas Ferreira; Tatiane Gomes Guedes; Sheila Coelho Ramalho Vasconcelos Morais; José Cristovam Martins Vieira; Marcelle Guimarães de Mello; Francisca Márcia Pereira Linhares

    2016-01-01

    ...: the diagnoses are related to factors that compromise the biopsychosocial health. The nurse, health staff member in the prison setting, must recognize and assess the individual and collective needs of women deprived of freedom. The inference of nursing diagnoses, based on clinical reasoning, contributes to a humanized, empathic and special care.

  9. 17 CFR 240.17g-3 - Annual financial reports to be furnished by nationally recognized statistical rating organizations.

    Science.gov (United States)

    2010-04-01

    ... financial statements of the nationally recognized statistical rating organization or audited consolidated financial statements of its parent if the nationally recognized statistical rating organization is a...) of this section are consolidated financial statements of the parent of the nationally recognized...

  10. Recognizing Symptoms

    Science.gov (United States)

    ... and how improve living with these conditions . Publication Library How Your Digestive System Works Words to Know About IFFGD About IFFGD Our Mission Awareness Activities Advocacy Activities Research Leadership Industry Council Contact us IBS Treatment Working With ...

  11. Recognizing Hypothermia

    Centers for Disease Control (CDC) Podcasts

    2007-11-01

    Hypothermia is a serious medical condition that strikes during very cold weather or when people are chilled from rain, sweat, or cold water.  Created: 11/1/2007 by Emergency Communications System.   Date Released: 12/13/2007.

  12. Recognizing resilience

    Science.gov (United States)

    Erika S. Svendsen; Gillian Baine; Mary E. Northridge; Lindsay K. Campbell; Sara S. Metcalf

    2014-01-01

    In 2012, a year after a devastating tornado hit the town of Joplin, Missouri, leaving 161 people dead and leveling Joplin High School and St. John's Hospital, President Obama addressed the graduating seniors: "There are a lot of stories here in Joplin of unthinkable courage and resilience. . . . [People in Joplin] learned that we have the power to...

  13. Perspectives of Women Decision-Makers Over the Participation and Recreational Events in Sports: A Turkish Perspective

    Science.gov (United States)

    Guzel, Pinar

    2015-01-01

    The purpose of this research is to put forth the role of the leisure and recreation events awareness including women decision-makers effects on their fellow women. Three main themes were recognized: Past; "Process of leisure and recreation events of women in Turkey", Present; "Model of Turkey for women on leisure and…

  14. Envisioning Women-Centered HIV Care: Perspectives from Women Living with HIV in Canada.

    Science.gov (United States)

    O'Brien, Nadia; Greene, Saara; Carter, Allison; Lewis, Johanna; Nicholson, Valerie; Kwaramba, Gladys; Ménard, Brigitte; Kaufman, Elaina; Ennabil, Nourane; Andersson, Neil; Loutfy, Mona; de Pokomandy, Alexandra; Kaida, Angela

    Women comprise nearly one-quarter of people living with human immunodeficiency virus (HIV) in Canada. Compared with men, women living with HIV experience inequities in HIV care and health outcomes, prompting a need for gendered and tailored approaches to HIV care. Peer and academic researchers from the Canadian HIV Women's Sexual and Reproductive Health Cohort Study conducted focus groups to understand women's experience of seeking care, with the purpose of identifying key characteristics that define a women-centered approach to HIV care. Eleven focus groups were conducted with 77 women living with HIV across Quebec, Ontario, and British Columbia, Canada. Women envisioned three central characteristics of women-centered HIV care, including i) coordinated and integrated services that address both HIV and women's health care priorities, and protect against exclusion from care due to HIV-related stigma, ii) care that recognizes and responds to structural barriers that limit women's access to care, such as violence, poverty, motherhood, HIV-related stigma, and challenges to safe disclosure, and iii) care that fosters peer support and peer leadership in its design and delivery to honor the diversity of women's experiences, overcome women's isolation, and prioritize women's ownership over the decisions that affect their lives. Despite advances in HIV treatment and care, the current care landscape is inadequate to meet women's comprehensive care needs. A women-centered approach to HIV care, as envisioned by women living with HIV, is central to guiding policy and practice to improve care and outcomes for women living with HIV in Canada. Copyright © 2017 Jacobs Institute of Women's Health. Published by Elsevier Inc. All rights reserved.

  15. Women's rights are human rights.

    Science.gov (United States)

    Shalala, D E

    1998-09-01

    The US Secretary of Health and Human Services, Donna Shalala, challenged the world to live up to the affirmation made in Cairo: that women's rights are human rights. The US has responded to this affirmation with vigor by recognizing that when given knowledge, education, opportunity, and power, women can be heroines; they can move mountains to help themselves and the others whom they are destined to nurture. The US Department of Health and Human Services (DHHS) has implemented numerous programs that will support these heroic acts. These programs include the Girl Power campaign (designed to help girls through the critical period of pre-adolescence), the National Strategy to Prevent Teen Pregnancy (promotes education to encourage abstinence), the National Plan on Breast Cancer, the Breast and Cervical Cancer Screening Program (protects the health of low-income women), the Women's Health Initiative at the National Institutes of Health (designed to increase our knowledge about hormone replacement therapy, dietary patterns, and exercise), and a national 24-hour toll-free Violence Against Women hotline (1-800-799-SAFE). Although DHHS has done much in 5 years, the US has a long way to go in dealing with the interconnecting issues of the human rights of women everywhere.

  16. Women as Mendelians and Geneticists

    Science.gov (United States)

    Richmond, Marsha L.

    2015-01-01

    After the rediscovery of Mendel's laws of heredity in 1900, the biologists who began studying heredity, variation, and evolution using the new Mendelian methodology—performing controlled hybrid crosses and statistically analyzing progeny to note the factorial basis of characters—made great progress. By 1910, the validity of Mendelism was widely recognized and the field William Bateson christened `genetics' was complemented by the chromosome theory of heredity of T. H. Morgan and his group in the United States. Historians, however, have largely overlooked an important factor in the early establishment of Mendelism and genetics: the large number of women who contributed to the various research groups. This article examines the social, economic, and disciplinary context behind this new wave of women's participation in science and describes the work of women Mendelians and geneticists employed at three leading experimental research institutes, 1900-1940. It argues that the key to more women working in science was the access to higher education and the receptivity of emerging interdisciplinary fields such as genetics to utilize the expertise of women workers, which not only advanced the discipline but also provided new opportunities for women's employment in science.

  17. Can Independent Judges Recognize Different Psychotherapies? An Experience with Manual-Guided Therapies.

    Science.gov (United States)

    Luborsky, Lester; And Others

    1982-01-01

    Tested whether independent judges could recognize three different manual-guided psychotherapies, drug counseling, supportive-expressive psychotherapy, and cognitive-behavioral using a special rating form containing scales for the characteristic aspects of each type. Results indicated that manual-guided therapies can be reliably recognized.…

  18. Image Segmentation and Feature Extraction for Recognizing Strokes in Tennis Game Videos

    NARCIS (Netherlands)

    Zivkovic, Z.; van der Heijden, Ferdinand; Petkovic, M.; Jonker, Willem; Langendijk, R.L.; Heijnsdijk, J.W.J.; Pimentel, A.D.; Wilkinson, M.H.F.

    This paper addresses the problem of recognizing human actions from video. Particularly, the case of recognizing events in tennis game videos is analyzed. Driven by our domain knowledge, a robust player segmentation algorithm is developed for real video data. Further, we introduce a number of novel

  19. 77 FR 27833 - Requirements for Recognizing the Aviation and Aerospace Innovation in Science and Engineering Award

    Science.gov (United States)

    2012-05-11

    ... Office of the Secretary Requirements for Recognizing the Aviation and Aerospace Innovation in Science and Engineering Award AGENCY: Office of the Secretary of Transportation, U.S. Department of Transportation. ACTION: Notice of the announcement of Requirements for the Secretary of Transportation's RAISE (Recognizing...

  20. Self-incompatibilty in gamete recognition: single self-recognizing determinants and multiple, non-self-recognizing ones function in the same individual.

    Science.gov (United States)

    Harada, Yoshito; Sawada, Hitoshi

    2013-01-01

    The frameworks (key mechanisms) of the self/non-self-discrimination systems that are found in various organisms have not been actively selected for, but have evolved by genetic drift such that the genetic frequency of random, advantageous mutations has increased within the genomes of these species by natural selection. The passive nature of this process leads to an important conclusion: in the self/non-self-discrimination system, the number of self-recognizing determinants becomes one compared to multiple non-self-recognizing determinants. Thus, the number of determinants is defined not by the character of the determinant, but by the system framework. Copyright © 2012 Wiley Periodicals, Inc.

  1. Autosomal-Recessive Posterior Microphthalmos Is Caused by Mutations in PRSS56, a Gene Encoding a Trypsin-Like Serine Protease

    Science.gov (United States)

    Gal, Andreas; Rau, Isabella; El Matri, Leila; Kreienkamp, Hans-Jürgen; Fehr, Susanne; Baklouti, Karim; Chouchane, Ibtissem; Li, Yun; Rehbein, Monika; Fuchs, Josefine; Fledelius, Hans C.; Vilhelmsen, Kaj; Schorderet, Daniel F.; Munier, Francis L.; Ostergaard, Elsebet; Thompson, Debra A.; Rosenberg, Thomas

    2011-01-01

    Posterior microphthalmos (MCOP) is a rare isolated developmental anomaly of the eye characterized by extreme hyperopia due to short axial length. The population of the Faroe Islands shows a high prevalence of an autosomal-recessive form (arMCOP) of the disease. Based on published linkage data, we refined the position of the disease locus (MCOP6) in an interval of 250 kb in chromosome 2q37.1 in two large Faroese families. We detected three different mutations in PRSS56. Patients of the Faroese families were either homozygous for c.926G>C (p.Trp309Ser) or compound heterozygous for c.926G>C and c.526C>G (p.Arg176Gly), whereas a homozygous 1 bp duplication (c.1066dupC) was identified in five patients with arMCOP from a consanguineous Tunisian family. In one patient with MCOP from the Faroe Islands and in another one from Turkey, no PRSS56 mutation was detected, suggesting nonallelic heterogeneity of the trait. Using RT-PCR, PRSS56 transcripts were detected in samples derived from the human adult retina, cornea, sclera, and optic nerve. The expression of the mouse ortholog could be first detected in the eye at E17 and was maintained into adulthood. The predicted PRSS56 protein is a 603 amino acid long secreted trypsin-like serine peptidase. The c.1066dupC is likely to result in a functional null allele, whereas the two point mutations predict the replacement of evolutionary conserved and functionally important residues. Molecular modeling of the p.Trp309Ser mutant suggests that both the affinity and reactivity of the enzyme toward in vivo protein substrates are likely to be substantially reduced. PMID:21397065

  2. Effects of trypsin inhibitor and particle size of expeller-extracted soybean meal on broiler live performance and weight of gizzard and pancreas.

    Science.gov (United States)

    Pacheco, W J; Stark, C R; Ferket, P R; Brake, J

    2014-09-01

    Soybean meal (SBM) is a co-product that remains after the oil is removed from whole soybeans by either solvent extraction or mechanically pressing soybeans using an expeller. Expeller-extracted soybean meal (ESBM) contains more fat and less protein than solvent-extracted soybean meal (SSBM), but has higher trypsin inhibitors (TI) that limit its inclusion in poultry diets. We hypothesized that increasing the particle size of ESBM could enhance chick tolerance to dietary TI. The experiment consisted of a 2 × 6 factorial arrangement of 2 ESBM particle sizes (coarse, 1,300 µm, and fine, 530 µm) and 6 TI levels (6, 9, 12, 15, 18, and 21 TIU/mg). A total of 672 male broiler chicks was randomly assigned among the 12 treatment combinations with 8 replicates per combination. The birds were provided with 1 kg/bird of starter diet in a crumble form. The pancreas and gizzard were excised and weighed at 16 d of age and organ weights relative to BW calculated. Feed consumption, BW, and feed conversion ratio (FCR) were determined at 7 and 14 d of age. Increasing TI level produced a quadratic effect (P < 0.01) on BW and FCR at 14 d of age. The best performance was obtained when chicks were fed diets containing ESBM at intermediate TI levels. Increasing the particle size of the ESBM from 530 to 1,300 µm improved BW and feed intake (P < 0.01) without affecting FCR. The relative weight of the pancreas increased linearly as TI level increased (P < 0.001). The results of this study indicated that chicks performed better when fed coarse ESBM at an intermediate TI level. © 2014 Poultry Science Association Inc.

  3. Structure and Dynamics of the Solvation of Bovine Pancreatic Trypsin Inhibitor in Explicit Water: A Comparative Study of the Effects of Solvent and Protein Polarizability

    Science.gov (United States)

    Kim, Byungchan; Young, Tom; Harder, Edward; Friesner, Richard A.; Berne, B. J.

    2009-01-01

    To isolate the effects of the inclusion of polarizability in the force field model on the structure and dynamics of the solvating water in differing electrostatic environments of proteins, we present the results of molecular dynamics simulations of the bovine pancreatic trypsin inhibitor (BPTI) in water with force fields that explicitly include polarization for both the protein and the water. We use three model potentials for water and two model potentials for the protein. Two of the water models and one of the protein models are polarizable. A total of six systems were simulated representing all combinations of these polarizable and nonpolarizable protein and water force fields. We find that all six systems behave in a similar manner in regions of the protein that are weakly electrostatic (either hydrophobic or weakly hydrophilic). However, in the vicinity of regions of the protein with relatively strong electrostatic fields (near positively or negatively charged residues), we observe that the water structure and dynamics are dependent on both the model of the protein and the model of the water. We find that a large part of the dynamical dependence can be described by small changes in the local environments of each region that limit the local density of non-hydrogen-bonded waters, precisely the water molecules that facilitate the dynamical relaxation of the water–water hydrogen bonds. We introduce a simple method for rescaling for this effect. When this is done, we are able to effectively isolate the influence of polarizability on the dynamics. We find that the solvating water’s relaxation is most affected when both the protein and the water models are polarizable. However, when only one model (or neither) is polarizable, the relaxation is similar regardless of the models used. PMID:16853101

  4. Simple and rapid LC-MS/MS method for the absolute determination of cetuximab in human serum using an immobilized trypsin.

    Science.gov (United States)

    Shibata, Kaito; Naito, Takafumi; Okamura, Jun; Hosokawa, Seiji; Mineta, Hiroyuki; Kawakami, Junichi

    2017-11-30

    Proteomic approaches using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) without an immunopurification technique have not been applied to the determination of serum cetuximab. This study developed a simple and rapid LC-MS/MS method for the absolute determination of cetuximab in human serum and applied it to clinical settings. Surrogate peptides derived from cetuximab digests were selected using a Fourier transform mass spectrometer. Reduced-alkylated serum cetuximab without immunopurification was digested for 20minutes using immobilized trypsin, and the digestion products were purified by solid-phase extraction. The LC-MS/MS was run in positive ion multiple reaction monitoring mode. This method was applied to the determination of serum samples in head and neck cancer patients treated with cetuximab. The chromatographic run time was 10minutes and no peaks interfering with surrogate peptides in serum digestion products were observed. The calibration curve of absolute cetuximab in serum was linear over the concentration range of 4-200μg/mL. The lower limit of quantification of cetuximab in human serum was 4μg/mL. The intra-assay and inter-assay precision and accuracy were less than 13.2% and 88.0-100.7%, respectively. The serum concentration range of cetuximab was 19-140μg/mL in patients. The serum cetuximab concentrations in LC-MS/MS were correlated with those in ELISA (r=0.899, P <0.01) and the mean bias was 1.5% in cancer patients. In conclusion, the present simple and rapid method with acceptable analytical performance can be helpful for evaluating the absolute concentration of serum cetuximab in clinical settings. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Effects of radiation therapy on tissue and serum concentrations of tumour associated trypsin inhibitor and their prognostic significance in rectal cancer patients

    Directory of Open Access Journals (Sweden)

    Stenman Ulf-Håkan

    2011-08-01

    Full Text Available Abstract Background We have previously demonstrated that elevated concentrations of tumour-associated trypsin inhibitor (TATI in both tumour tissue (t-TATI and in serum (s-TATI are associated with a poor prognosis in colorectal cancer patients. It was also found that s-TATI concentrations were lower in patients with rectal cancer compared to patients with colon cancer. In this study, we investigated the effects of neoadjuvant radiotherapy (RT on concentrations of t-TATI and s-TATI in patients with rectal cancer. Methods TATI was analysed in serum, normal mucosa and tumour tissue collected at various time points in 53 rectal cancer patients enrolled in a case-control study where 12 patients received surgery alone, 20 patients 5 × 5 Gy (short-term preoperative RT and 21 patients 25 × 2 Gy (long-term preoperative RT. T-TATI was analysed by immunohistochemistry and s-TATI was determined by an immunofluorometric assay. Mann-Whitney U test and Wilcoxon Z (Z test were used to assess t-TATI and s-TATI concentrations in relation to RT. Spearman's correlation (R test was used to explore the associations between t-TATI, s-TATI and clinicopathological parameters. Overall survival (OS according to high and low t-TATI and s-TATI concentrations was estimated by classification and regression tree analysis, Kaplan-Meier analysis and the log rank test. Results RT did not affect concentrations of t-TATI or s-TATI. In patients receiving short-term but not long-term RT, s-TATI concentrations were significantly higher 4 weeks post surgery than in serum drawn prior to surgery (Z = -3.366, P Conclusions The results presented here further validate the utility of t-TATI and s-TATI as prognostic biomarkers in patients with rectal cancer, independent of neoadjuvant RT.

  6. Urea free and more efficient sample preparation method for mass spectrometry based protein identification via combining the formic acid-assisted chemical cleavage and trypsin digestion.

    Science.gov (United States)

    Wu, Shuaibin; Yang, Kaiguang; Liang, Zhen; Zhang, Lihua; Zhang, Yukui

    2011-10-30

    A formic acid (FA)-assisted sample preparation method was presented for protein identification via mass spectrometry (MS). Detailedly, an aqueous solution containing 2% FA and dithiothreitol was selected to perform protein denaturation, aspartic acid (D) sites cleavage and disulfide linkages reduction simultaneously at 108°C for 2h. Subsequently, FA wiped off via vacuum concentration. Finally, iodoacetamide (IAA) alkylation and trypsin digestion could be performed ordinally. A series of model proteins (BSA, β-lactoglobulin and apo-Transferrin) were treated respectively using such method, followed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) analysis. The identified peptide number was increased by ∼ 80% in comparison with the conventional urea-assisted sample preparation method. Moreover, BSA identification was achieved efficiently down to femtomole (25 ± 0 sequence coverage and 16 ± 1 peptides) via such method. In contrast, there were not peptides identified confidently via the urea-assisted method before desalination via the C18 zip tip. The absence of urea in this sample preparation method was an advantage for the more favorable digestion and MALDI-TOF MS analysis. The performances of two methods for the real sample (rat liver proteome) were also compared, followed by a nanoflow reversed-phase liquid chromatography with electrospray ionization tandem mass spectrometry system analysis. As a result, 1335 ± 43 peptides were identified confidently (false discovery rate method, corresponding to 295 ± 12 proteins (of top match=1 and requiring 2 unique peptides at least). In contrast, there were only 1107 ± 16 peptides (corresponding to 231 ± 10 proteins) obtained from the conventional urea-assisted method. It was serving as a more efficient protein sample preparation method for researching specific proteomes better, and providing assistance to develop other proteomics analysis methods, such as, peptide

  7. Expression of trypsin modulating oostatic factor (TMOF) in an entomopathogenic fungus increases its virulence towards Anopheles gambiae and reduces fecundity in the target mosquito.

    Science.gov (United States)

    Kamareddine, Layla; Fan, Yanhua; Osta, Mike A; Keyhani, Nemat O

    2013-01-21

    Adult and larval mosquitoes regulate food digestion in their gut with trypsin modulating oostatic factor (TMOF), a decapeptide hormone synthesized by the ovaries and the neuroendocrine system. TMOF is currently being developed as a mosquitocide, however, delivery of the peptide to the mosquito remains a significant challenge. Entomopathogenic fungi offer a means for targeting mosquitoes with TMOF. The efficacy of wild type and transgenic Beauveria bassiana strains expressing Aedes aegypti TMOF (Bb-Aa1) were evaluated against larvae and sugar- and blood-fed adult Anopheles gambiae mosquitoes using insect bioassays. Bb-Aa1 displayed increased virulence against larvae, and sugar and blood fed adult A. gambiae when compared to the wild type parent strain. Median lethal dose (LD50) values decreased by ~20% for larvae, and ~40% for both sugar and blood-fed mosquitoes using Bb-Aa1 relative to the wild type parent. Median lethal time (LT50) values were lower for blood-fed compared to sugar-fed mosquitoes in infections with both wild type and Bb-Aa1. However, infection using Bb-Aa1 resulted in 15% to 25% reduction in LT50 values for sugar- and blood fed mosquitoes, and ~27% for larvae, respectively, relative to the wild type parent. In addition, infection with Bb-Aa1 resulted in a dramatic reduction in fecundity of the target mosquitoes. B. bassiana expressing Ae. aegypti TMOF exhibited increased virulence against A. gambiae compared to the wild type strain. These data expand the range and utility of entomopathogenic fungi expressing mosquito-specific molecules to improve their biological control activities against mosquito vectors of disease.

  8. Expression of trypsin modulating oostatic factor (TMOF in an entomopathogenic fungus increases its virulence towards Anopheles gambiae and reduces fecundity in the target mosquito

    Directory of Open Access Journals (Sweden)

    Kamareddine Layla

    2013-01-01

    Full Text Available Abstract Background Adult and larval mosquitoes regulate food digestion in their gut with trypsin modulating oostatic factor (TMOF, a decapeptide hormone synthesized by the ovaries and the neuroendocrine system. TMOF is currently being developed as a mosquitocide, however, delivery of the peptide to the mosquito remains a significant challenge. Entomopathogenic fungi offer a means for targeting mosquitoes with TMOF. Findings The efficacy of wild type and transgenic Beauveria bassiana strains expressing Aedes aegypti TMOF (Bb-Aa1 were evaluated against larvae and sugar- and blood-fed adult Anopheles gambiae mosquitoes using insect bioassays. Bb-Aa1 displayed increased virulence against larvae, and sugar and blood fed adult A. gambiae when compared to the wild type parent strain. Median lethal dose (LD50 values decreased by ~20% for larvae, and ~40% for both sugar and blood-fed mosquitoes using Bb-Aa1 relative to the wild type parent. Median lethal time (LT50 values were lower for blood-fed compared to sugar-fed mosquitoes in infections with both wild type and Bb-Aa1. However, infection using Bb-Aa1 resulted in 15% to 25% reduction in LT50 values for sugar- and blood fed mosquitoes, and ~27% for larvae, respectively, relative to the wild type parent. In addition, infection with Bb-Aa1 resulted in a dramatic reduction in fecundity of the target mosquitoes. Conclusions B. bassiana expressing Ae. aegypti TMOF exhibited increased virulence against A. gambiae compared to the wild type strain. These data expand the range and utility of entomopathogenic fungi expressing mosquito-specific molecules to improve their biological control activities against mosquito vectors of disease.

  9. A Preclinical Translational Study of the Cardioprotective Effects of Plasma-Derived Alpha-1 Anti-trypsin in Acute Myocardial Infarction.

    Science.gov (United States)

    Mauro, Adolfo G; Mezzaroma, Eleonora; Marchetti, Carlo; Narayan, Pratyush; Del Buono, Marco G; Capuano, Marialessia; Prestamburgo, Andrea; Catapano, Simone; Salloum, Fadi N; Abbate, Antonio; Toldo, Stefano

    2017-05-01

    The area of myocardial infarction continues to expand for hours after reperfusion. The injured but viable myocardium may be salvaged if the signals leading to cell death are interrupted. Activation of the caspase-1 inflammasome in the heart shortly after ischemia-reperfusion contributes to the final infarct size. Plasma-derived α-1 anti-trypsin (AAT) has shown to inhibit inflammasome formation in vitro and in vivo. To explore the potential translational clinical value of AAT as a therapeutic, we conducted a series of preclinical experiments designed to simulate clinically relevant scenarios. Adult male CD1 mice were used. The left anterior descending coronary artery was ligated for 30 or 75 minutes followed by reperfusion, to explore different severity of ischemic injury. Plasma-derived AAT (Prolastin C) was administered intraperitoneally after reperfusion, without pretreatment, exploring 3 different doses (60, 120, and 180 mg/kg). In a subgroup of mice, we administered Prolastin C with a delay of 30 minutes after reperfusion to simulate the clinical context of delayed administration, and we also used a model of permanent coronary artery ligation without reperfusion. Finally, we tested whether a single dose at reperfusion was sufficient to maintain a benefit in the longer term (7 days). Infarct size was measured by 3 different and independent methodologies: pathology, plasma levels of troponin I, and wall motion abnormalities at echocardiography. Prolastin C given at reperfusion after 30 minutes of ischemia provided a powerful reduction in infarct size (>50% reduction in all methodology used, all P infarct size. Plasma-derived AAT (Prolastin C) given as an adjunct to reperfusion powerfully limits the final infarct size across a wide range of experiments in the mouse reproducing clinically relevant scenarios, such as variable duration of ischemia, delay in administration in the drug, and a large therapeutic index.

  10. Recognition of voice commands using adaptation of foreign language speech recognizer via selection of phonetic transcriptions

    Science.gov (United States)

    Maskeliunas, Rytis; Rudzionis, Vytautas

    2011-06-01

    In recent years various commercial speech recognizers have become available. These recognizers provide the possibility to develop applications incorporating various speech recognition techniques easily and quickly. All of these commercial recognizers are typically targeted to widely spoken languages having large market potential; however, it may be possible to adapt available commercial recognizers for use in environments where less widely spoken languages are used. Since most commercial recognition engines are closed systems the single avenue for the adaptation is to try set ways for the selection of proper phonetic transcription methods between the two languages. This paper deals with the methods to find the phonetic transcriptions for Lithuanian voice commands to be recognized using English speech engines. The experimental evaluation showed that it is possible to find phonetic transcriptions that will enable the recognition of Lithuanian voice commands with recognition accuracy of over 90%.

  11. Evaluation of speech recognizers for use in advanced combat helicopter crew station research and development

    Science.gov (United States)

    Simpson, Carol A.

    1990-01-01

    The U.S. Army Crew Station Research and Development Facility uses vintage 1984 speech recognizers. An evaluation was performed of newer off-the-shelf speech recognition devices to determine whether newer technology performance and capabilities are substantially better than that of the Army's current speech recognizers. The Phonetic Discrimination (PD-100) Test was used to compare recognizer performance in two ambient noise conditions: quiet office and helicopter noise. Test tokens were spoken by males and females and in isolated-word and connected-work mode. Better overall recognition accuracy was obtained from the newer recognizers. Recognizer capabilities needed to support the development of human factors design requirements for speech command systems in advanced combat helicopters are listed.

  12. Mining Emerging Patterns for Recognizing Activities of Multiple Users in Pervasive Computing

    DEFF Research Database (Denmark)

    Gu, Tao; Wu, Zhanqing; Wang, Liang

    2009-01-01

    Understanding and recognizing human activities from sensor readings is an important task in pervasive computing. Existing work on activity recognition mainly focuses on recognizing activities for a single user in a smart home environment. However, in real life, there are often multiple inhabitants...... activity models, and propose an Emerging Pattern based Multi-user Activity Recognizer (epMAR) to recognize both single-user and multiuser activities. We conduct our empirical studies by collecting real-world activity traces done by two volunteers over a period of two weeks in a smart home environment...... sensor readings in a home environment, and propose a novel pattern mining approach to recognize both single-user and multi-user activities in a unified solution. We exploit Emerging Pattern – a type of knowledge pattern that describes significant changes between classes of data – for constructing our...

  13. Women's health

    Science.gov (United States)

    ... may affect women can include: Bladder emptying disorders Urinary incontinence and overactive bladder Interstitial cystitis Prolapse of the ... the cervix (LEEP, Cone biopsy) Procedures to treat urinary incontinence Tubal ligation and reversal of tubal sterilization Uterine ...

  14. pregnant women

    Directory of Open Access Journals (Sweden)

    Ewelina Buraczewska

    2016-09-01

    Full Text Available Background . Diabetes in pregnancy is a condition which includes pre-pregnancy diabetes in women already suffering from diabetes who become pregnant, and hyperglycaemia first diagnosed during pregnancy, defined as a disorder of carbohydrate tolerance resulting in increased blood glucose concentrations, which were first diagnosed in pregnant healthy women. Women’s knowledge about the disease and the practical use of this knowledge play an important role in the healing process. Objectives. The assessment of the state of knowledge about diabetes in pregnant women diagnosed with hyperglycaemia during pregnancy. Material and methods. The study involved 127 pregnant women with hyperglycemia which was first diagnosed during pregnancy. The median age of the subjects was 32.1 (19–45. A diagnostic survey was a research method. A self-prepared survey questionnaire was the research tool. The results were statistically analyzed. Results. The surveyed women assessed their knowledge about gestational diabetes as good and very good. The study showed, however, very poor knowledge about the clinical signs of diabetes among the subjects. The most frequently cited risk factors for gestational diabetes include: the presence of diabetes in one’s family, overweight states or obesity, and diabetes in previous pregnancy. The Internet was the main source of knowledge about diabetes among the subjects. Conclusions . 1. The level of knowledge in women about gestational diabetes is insufficient. 2. There is a need to extend educational activities related to the symptoms of diabetes and the principles of its prevention among pregnant women. 3. The participation of general practitioners in the education of women about diabetes in pregnancy is insufficient.

  15. [Is coronary artery disease different in women?].

    Science.gov (United States)

    Schiele, François; Chopard, Romain

    2014-01-01

    Coronary artery disease (CAD) is the primary cause of death in women. Although acute coronary syndrome (ACS) is relatively infrequent in young women, failure to recognize ACS in this population can incur a major risk and registry data show that there is still plenty of room for improvement in this area. Women may suffer from "classical" CAD with development of atherosclerosis with a delay of about 10 years as compared to men, reflecting hormonal protection in women. Besides this classical presentation, angina in women often corresponds to impaired microcirculation, a syndrome known to associate typical angina, demonstrable myocardial ischemia, but no lesions on the coronary angiography. Finally, spasm, spontaneous dissection or coronary thrombosis through endothelial rupture are more frequent in women. The influence of risk factors on the development of CAD is comparable in both women and men. Recent registry studies show that in France, in particular, diabetes, obesity, and smoking are all risk factors that are on the rise in women. In addition, certain other risk factors are more specific to women, namely psycho-social stress. The methods to evaluate risk and detect CAD were mainly developed in male study populations, and these tools thus perform less well in female patients. In case of ACS, women benefit just as much from invasive management, but are at greater risk of iatrogenic complications, particularly with anti-thrombotic therapy or during revascularization procedures. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  16. Intimate Partner Violence against Older Women in Germany: Prevalence and Associated Factors

    Science.gov (United States)

    Stockl, Heidi; Watts, Charlotte; Penhale, Bridget

    2012-01-01

    Violence against women is a recognized human rights and public health issue, with significant impacts on women's life and health. Until now, several studies, most of them relying on small scale samples, have explored the prevalence and health effects of intimate partner violence against older women, whereas few have examined what actually puts…

  17. Kenyan Girls as Agents of Peace: Enhancing the Capacity of Future Women Peacebuilders

    Science.gov (United States)

    Zanoni, Katie

    2017-01-01

    The role of women in peacebuilding efforts has been recognized through various international instruments that have advanced the ability of women to access the peace table. In order for women to act as leaders, they must possess the capacity to disrupt structural, cultural, and direct forms of violence, engage in peacemaking activities, and employ…

  18. The Extent of Off-Farm Employment and Its Impact on Farm Women.

    Science.gov (United States)

    Danes, Sharon M.; Keskinen, Susan M.

    1990-01-01

    Surveys off-farm employment of farm women and its impact on their lives. Describes survey data illustrating financial hardships experienced by working farm women. Describes imbalance between work and leisure as major source of dissatisfaction. Suggests service providers recognize women's "role overload" and its effects on families. (TES)

  19. Recruiting women smokers: the engineering of consent.

    Science.gov (United States)

    Brandt, A M

    1996-01-01

    A range of social forces contributed to the effective recruitment of women to cigarette smoking in the crucial period between 1900 and 1940. Cigarette advertisers and public relations experts recognized the significance of women's changing roles and the rising culture of consumption, and worked to create specific meanings for the cigarette to make it appeal to women. The cigarette was a flexible symbol, with a remarkably elastic set of meanings; for women, it represented rebellious independence, glamour, seduction, and sexual allure, and served as a symbol for both feminists and flappers. The industry, with the help of advertisers and public relations experts, effectively engineered consent for women as smokers. The "engineering of consent" has a role to play in smoking cessation, since negative meanings for the cigarette can be engineered as well.

  20. Recognizing Multi-user Activities using Wearable Sensors in a Smart Home

    DEFF Research Database (Denmark)

    Wang, Liang; Gu, Tao; Tao, Xianping

    2010-01-01

    The advances of wearable sensors and wireless networks oer many opportunities to recognize human activities from sensor readings in pervasive computing. Existing work so far focuses mainly on recognizing activities of a single user in a home environment. However, there are typically multiple...... inhabitants in a real home and they often perform activities together. In this paper, we investigate the problem of recognizing multi-user activities using wearable sensors in a home setting. We develop a multi-modal, wearable sensor platform to collect sensor data for multiple users, and study two temporal...

  1. Women and Diabetes

    Medline Plus

    Full Text Available ... Mammography Women and Diabetes HPV, HIV, Birth Control Heart Health for Women Pregnancy Menopause More Women's Health Topics Resources for You YourDiabetesInfo.org American Diabetes Association Get Other FDA Publications for Women For Women ...

  2. On Campus with Women.

    Science.gov (United States)

    Liberal Education, 1988

    1988-01-01

    A newsletter on women and higher education contains a variety of articles on women and sports, women's studies, working in academe, minority women, campus life and environment, child care, international education, women and science, sexual harassment, and campus violence. (MSE)

  3. Women and Diabetes

    Medline Plus

    Full Text Available ... Information by Audience For Women Women's Health Topics Women and Diabetes Share Tweet Linkedin Pin it More ... women during pregnancy. Diabetes and Pregnancy (CDC) Diverse Women in Clinical Trials Campaign Clinical trials can help ...

  4. Women and Diabetes

    Medline Plus

    Full Text Available ... Office of Research on Women's Health to raise awareness about diverse women of different ages, races, ethnic ... Women's Health Topics Mammography Women and Diabetes HPV, HIV, Birth Control Heart Health for Women Pregnancy Menopause ...

  5. Next Generation Luminaires: Recognizing Innovative, Energy-Efficient Commercial Lighting Luminaires

    Energy Technology Data Exchange (ETDEWEB)

    None

    2013-04-01

    Fact sheet that describes the Next Generation Luminaires SSL lighting design competition, which recognizes excellence in technical innovation and design of high-quality, energy-efficient commercial lighting, both indoor and outdoor.

  6. Can You Recognize a Heart Attack or Stroke? What To Do When Every Moment Counts

    Science.gov (United States)

    ... Special Issues Subscribe August 2014 Print this issue Can You Recognize a Heart Attack or Stroke? What ... could prevent many of these deaths. Fast action can also limit permanent damage to the body. Heart ...

  7. EPA Recognizes Whole Foods Market in Marietta, Ga for Reducing Food Waste

    Science.gov (United States)

    ATLANTA - Today, U.S. Environmental Protection Agency (EPA) Regional Administrator Heather McTeer Toney recognized the Whole Foods Market Merchant Walk Marietta Store in Georgia for the store's achievements in EPA's Food Recovery Challenge. Whole Fo

  8. Two New England Businesses and One Individual Recognized by EPA for Climate Leadership

    Science.gov (United States)

    EPA has recognized the leadership of one individual and two New England companies for their climate actions, such as investing in energy efficiency and renewable energy, and preparing for the impacts of climate change.

  9. Realistic Creativity Training for Innovation Practitioners: The Know-Recognize-React Model

    DEFF Research Database (Denmark)

    Valgeirsdóttir, Dagný; Onarheim, Balder

    2017-01-01

    As creativity becomes increasingly recognized as important raw material for innovation, the importance of identifying ways to increase practitioners’ creativity through rigorously designed creativity training programs is highlighted. Therefore we sat out to design a creativity training program sp...

  10. Measurement of the ability of science students to recognize business opportunities

    NARCIS (Netherlands)

    Nab, J.; Oost, H.; Pilot, A.; van Keulen, H.

    2008-01-01

    This paper describes the development of an instrument measuring students’ ability to recognize business opportunities. Recognition of business opportunities where others do not is one of the basic qualities of entrepreneurs, and therefore needs attention in entrepreneurship education. However, only

  11. The Linguistic Model in Biology: Implications for Recognizing Life and Intelligence

    Science.gov (United States)

    Waters, D. P.

    2010-04-01

    Early molecular biologists relied on a "linguistic metaphor," still heard in terms like "transcription" and "translation." This deeply-rooted metaphor may help astrobiologists understand how to recognize forms of life elsewhere in the universe.

  12. Inter-alpha-trypsin inhibitor heavy chain 4: a novel biomarker for environmental exposure to particulate air pollution in patients with chronic obstructive pulmonary disease

    Directory of Open Access Journals (Sweden)

    Lee KY

    2015-04-01

    Full Text Available Kang-Yun Lee,1–3 Po-Hao Feng,1,2 Shu-Chuan Ho,4 Kai-Jen Chuang,5,6 Tzu-Tao Chen,2,3 Chien-Ling Su,2,4 Wen-Te Liu,2,4 Hsiao-Chi Chuang2,4 1Department of Internal Medicine, School of Medicine, College of Medicine, Taipei Medical University, 2Division of Pulmonary Medicine, Department of Internal Medicine, Shuang Ho Hospital, Taipei Medical University, 3Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, 4School of Respiratory Therapy, College of Medicine, Taipei Medical University, 5Department of Public Health, School of Medicine, College of Medicine, Taipei Medical University, 6School of Public Health, College of Public Health and Nutrition, Taipei Medical University, Taipei, Taiwan Abstract: Chronic obstructive pulmonary disease (COPD is a chronic inflammatory disease that is correlated with environmental stress. Particulate matter ≤10 µm (PM10 is considered to be a risk factor for COPD development; however, the effects of PM10 on the protein levels in COPD remain unclear. Fifty subjects with COPD and 15 healthy controls were recruited. Gene ontology analysis of differentially expressed proteins identified immune system process and binding as the most important biological process and molecular function, respectively, in the responses of PM10-exposed patients with COPD. Biomarkers for PM10 in COPD were identified and compared with the same in healthy controls and included proteoglycan 4 (PRG4, inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4, and apolipoprotein F (APOF. PRG4 and ITIH4 were associated with a past 3-year PM10 exposure level. The receiver operating characteristic curve analysis showed that ITIH4 is a sensitive and specific biomarker for PM10 exposure (area under the curve [AUC] =0.690, P=0.015 compared with PRG4 (AUC =0.636, P=0.083, APOF (AUC =0.523, P=0.766, 8-isoprostane (AUC =0.563, P=0.405, and C-reactive protein (CRP; AUC =0.634, P=0.086. ITIH4 levels were correlated with CRP (r=0

  13. Efeito de taninos da lentilha sobre a hidrólise da albumina pela tripsina Effect of lentil tannins on albumin hydrolysis by trypsin

    Directory of Open Access Journals (Sweden)

    Valdir A. NEVES

    1997-12-01

    Full Text Available Os taninos da casca da semente de lentilha foram extraídos e purificados, levados à interação com albumina isolada de lentilha e com caseína; e estudados por turbidimetria. As interações da albumina e caseína com taninos purificados, a várias relações tanino-proteína, mostraram ser independente e dependente do pH, respectivamente. Hidrólise in vitro com tripsina das proteínas sem taninos indicou que o aquecimento a 99°C/15 min reduzia a susceptibilidade da albumina e aumentava a da caseína à tripsina. A influência de diferentes relações tanino:proteína (1:40; 1:20; 1:5; 1:2,5 na hidrólise mostrou maior inibição para caseína que para albumina de lentilha, independente de aquecimento. Após aquecimento ambas proteínas foram mais hidrolizadas para qualquer das relações tanino proteínas estudadas. A eletroforese em gel de poliacrilamida-dodecilsulfato de sódio do transcurso da hidrólise da interação tanino-albumina nativa mostra a dependência da relação tanino:proteína.The tannins of lentil seed coat were extracted and purified, allowed to interact with isolated lentil albumin and casein and studied turbidimetrically. The interactions of lentil albumin and casein with tannin at various tannin-to- protein ratios proved to be pH-independent and pH-dependent, respectively. In vitro trypsin hydrolysis of the proteins without tannins showed that the heating at 99°C/15 min. reduced the susceptibility of albumin and increased the susceptibility of casein to the enzyme. The influence of tannin-to-protein ratios (1:40; 1:20; 1:5. 1:2,5 on the in vitro tryptic hydrolysis was more inhibitory for casein than for lentil albumin when unheated or heated at 99°C/15 min. After heating both proteins were more hydrolyzed at all tannin-to-protein ratios. SDS-PAGE of the products of hydrolysis of native albumin-tannin interactions show that the extent of proteolysis is dependent of the protein-tannin ratio.

  14. Dietary probiotic supplementation improved gut amylase to trypsin ratio in European seabass reared at different temperatures and survival after handling stress

    Directory of Open Access Journals (Sweden)

    Luís Filipe Ferreira Pereira

    2014-07-01

    Full Text Available Antibiotics and chemical treatments are often used as disease control strategy. A prophylactic and alternative method to this chemical approach are the probiotics [1]. Probiotics are defined as "live microorganisms which when administered in adequate amounts confer a health benefit on the host" (FAO, 2001. The benefits of probiotic treatments are improvements in host nutritional retention [2], antagonistic properties to bacterial pathogen proliferation [3], modulation of immune responses [4], among others. Temperature plays a major role in dietary nutrient utilization and immune responses in fish, and have a modulatory effect on probiotic activity in intestine. The current study evaluated the use of a dietary probiotic supplementation in European seabass (Dicentrarchus labrax, one of the most important fish species in Southern Europe. Fish were fed on a multi-species probiotic (Bacillus sp., Pedicoccus sp., Enterococcus sp., Lactobacillus sp. , reared under 3 different temperatures (17, 20 and 23 ºC for 70 days. Fish were pair-fed, fixed to the voluntary feed intake of fish reared at 17 º C, in order to have similar probiotic intake among the temperature groups. Final body weight tripled initial weight (12.7 to 30.7g. At the end of the growth trial, all fish were subjected to a handling stress, in which stocking density increased by 6 fold (from 4 kg/m3 to 25 kg/m3, followed by a 15 min chase with a pole. Growth performance was not affected by the dietary treatment. Post-stress cumulative mortality were significantly higher in the 17ºC control group (figure 1A. Digestive enzymes activity were significantly affected by temperature and diet interaction. The activity ratio of amylase to trypsin (figure 1B increased with temperature and dietary probiotic supplementation, an indication that probiotic treatment at 23oC have a positive influence on the metabolic flexibility of carbohydrate-protein utilization. Non-specific immune response (ACH50

  15. Poly (N-acryloxysuccinimide-co-ethylene glycol dimethacrylate) precursor monolith and its post polymerization modification with alkyl ligands, trypsin and lectins for reversed-phase chromatography, miniaturized enzyme reactors and lectin affinity chromatography, respectively.

    Science.gov (United States)

    Jonnada, Murthy; El Rassi, Ziad

    2017-11-01

    This investigation was aimed at introducing a monolithic precursor that can be conveniently grafted with the desired chromatographic ligand via the process of post polymerization modification (PPM). The precursor was obtained by the in-situ polymerization of N-acryloxysuccinimide (NAS) and ethylene glycol dimethacrylate (EDMA) in a narrow bore stainless steel column of 1 mm i.d. yielding a poly(NAS-co-EDMA) monolithic column designated as the poly(NAS-co-EDMA) monolith (NASM) column. In a first PPM, the NASM column was bonded with octadecyl (OD) ligands yielding a nonpolar NASM-OD column that proved useful for reversed phase chromatography (RPC) of proteins in gradient elution at increasing %ACN in the mobile phase. NASM-OD resulted from the reaction between the N-hydroxysuccinimide of NASM with octadecyl amine. In a second PPM, NASM was surface immobilized with trypsin generating a proteolytic narrow bore enzyme reactor called NASM-trypsin immobilized enzyme reactor (IMER) that permitted the online digestion of proteins in a 20-min single pass through the IMER incorporated in a setup equipped with a short RPC column to achieve simultaneously a peptide tryptic map. This constituted a rapid turnover whereby ∼95% of the protein was hydrolyzed by the immobilized trypsin. In a third PPM, the NASM column was surface immobilized with three different lectins (LCA, Con A and RCA) having complementary affinities toward serum glycoproteins thus permitting the capture of a wide range of glycoproteins/glycoforms. The three NASM-lectin columns when operated in a tandem format led to assessing the level of the various glycoforms in human serum via LC-MS/MS analysis of the captured protein fractions by each NASM-lectin column. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Pancreatic secretory trypsin inhibitor causes autocrine-mediated migration and invasion in bladder cancer and phosphorylates the EGF receptor, Akt2 and Akt3, and ERK1 and ERK2.

    Science.gov (United States)

    Marchbank, Tania; Mahmood, Asif; Playford, Raymond J

    2013-08-01

    Pancreatic secretory trypsin inhibitor (PSTI) is expressed in most bladder carcinomas, where its pathophysiological relevance is unclear. Using recombinant normal sequence PSTI/tumor-associated trypsin inhibitor (TATI), a variant associated with familial pancreatitis (N34S), an active site-inactivated variant (R18/V19), and immunoneutralization and RNA interference-mediated knockdown techniques, we investigated the actions of PSTI/TATI on cell migration (wounding monolayers), collagen invasion (gel invasion assays), and proliferation (Alamar blue) on 253J, RT4, and HT1376 human bladder carcinoma cell lines. All three forms of PSTI/TATI stimulated migration twofold, and normal sequence PSTI/TATI showed synergistic promigratory effects when added with EGF. Addition of structurally unrelated soybean trypsin inhibitor had no promigratory activity. Similar results were seen using collagen invasion assays, although the active site mutated variant had no proinvasive activity, probably due to reduced Akt2 activation. PSTI/TATI did not stimulate proliferation despite acting, at least partially, through the EGF receptor, as effects of PSTI/TATI were truncated by the addition of an EGF receptor blocking antibody or the tyrosine kinase inhibitor tyrphostin. Cell lines produced endogenous PSTI/TATI, and PSTI/TATI RNA interference knockdown or the addition of PSTI/TATI, EGF receptor, or tyrphostin blocking agents reduced migration and invasion below baseline. PSTI/TATI induced phosphorylation of the EGF receptor, ERK1 and ERK2, Akt2 and Akt3, JNK1, MKK3, and ribosomal protein S6 kinase 1. This profile was more limited than that induced by EGF and did not include Akt1, probably explaining the lack of proproliferative activity. Our findings of autocrine stimulation and synergistic responses between EGF and PSTI/TATI at concentrations found in urine and tissue suggest that PSTI/TATI has pathophysiological relevance.

  17. Recognizing the needs – Student teachers´ learning to teach from teaching

    OpenAIRE

    Pernilla Nilsson

    2012-01-01

    This paper is based on an exploration of the ways in which primary science student teachers recognize and learn about issues that shape their own professional learning. The paper discusses different perspectives of “knowledgebase needed for teaching” and Shulman’s concept of pedagogical content knowledge, and explores how elements of knowledge are to be recognized and further developed within primary teacher education. Primary science student teacher participants (n = 25) were stimulated to u...

  18. Artificial cleavage site recognized by plum pox potyvirus protease in Escherichia coli.

    OpenAIRE

    García, J.A. (Juan Alonso); Riechmann, J L; S. Laín

    1989-01-01

    A synthetic plum pox virus (PPV) NIb-CP cleavage site was recognized by a PPV protease in an in vivo Escherichia coli expression system. The presence of the natural NIb-CP cleavage site did not affect processing at the artificial one. However, although both the proteases and the cleavage sites of PPV and tobacco etch virus show high sequence homology, a similar cassette from the tobacco etch virus NIb-CP junction was not efficiently recognized by the PPV protease.

  19. Smokefree Women

    Science.gov (United States)

    ... and advice about quitting smoking through real-time messaging. › HealthyYou TXT A text message program that provides ... share home about smokefree women for health professionals privacy accessibility viewing files disclaimer FOIA smokefree.gov teen. ...

  20. Women's Workshop.

    Science.gov (United States)

    Karelius, Karen

    The Women's Workshop Notebook is the tool used in the nine-week course designed for the mature woman returning to school at Antelope Valley College. The notebook exercises along with the group interaction and instruction stress the importance of personal assessment of strengths, weaknesses, dreams, deliberations and life history in…