Sample records for salivary amylase activity

  1. Influence of different restorative materials on lysozyme and amylase activity of the salivary pellicle in situ.

    PubMed

    Hannig, Christian; Wasser, Mathias; Becker, Klaus; Hannig, Matthias; Huber, Karin; Attin, Thomas

    2006-09-15

    Lysozyme and amylase are the most abundant enzymatic components in the salivary pellicle. The purpose of the present study was to determine the influence of different substrata on amylase and lysozyme activity in salivary pellicles formed in situ. Slabs (5 mm diameter) of bovine dentine and enamel, of titanium, gold alloy, resin composite, PMMA, amalgam, and feldspar ceramic were fixed on the buccal sites of individual splints worn by six subjects for 30 min to allow pellicle formation. Thereafter, slabs were removed from the trays and rinsed with running water. Lysozyme activity was determined via lysis of Micrococcus lysodeicticus. Amylase activity was measured with a photometric method using 2-chloro-4-nitrophenyl-4-O-beta-D-galactopyranosylmaltotriosid (GalG2CNP) as substrate. Both pellicle enzymes were evaluated in the immobilized as well as in the desorbed state. Salivary enzyme activities were also measured. All investigated pellicles exhibited lysozyme and amylase activity. Great intraindividual and interindividual differences were observed. Over all samples, immobilized amylase activity amounted to 0.65 +/- 0.64 mU/cm2. Immobilized lysozyme activity was 5.04 +/- 1.55 U/cm2. There were no major effects of the substratum on pellicle-bound amylase and lysozyme activity. Immobilized and desorbed enzyme activities revealed a strong correlation (lysozyme: r = 0.700; amylase: r = 0.990). Salivary enzyme activities had only little impact on pellicle-bound enzyme activities. Amylase and lysozyme are incorporated in the acquired in situ pellicle on different solid surfaces in an active conformation. Dental material and enzyme activity in the saliva have only little impact on enzymatic activity in the pellicle in situ. PMID:16739107

  2. Exercise upregulates salivary amylase in humans (Review)

    PubMed Central

    KOIBUCHI, ERI; SUZUKI, YOSHIO

    2014-01-01

    The secretion of salivary ?-amylase is influenced by adrenergic regulation of the sympathetic nervous system and the hypothalamic-pituitary-adrenal axis; thus, exercise affects the levels of salivary ?-amylase. Granger et al published a review in 2007 that focused attention on salivary ?-amylase. In addition, a portable system for monitoring salivary ?-amylase activity was launched in Japan at the end of 2005. The correlation between exercise and salivary ?-amylase has since been extensively investigated. The present review summarizes relevant studies published in the English and Japanese literature after 2006. A search of the PubMed and CiNii databases identified 54 articles, from which 15 original articles were selected. The findings described in these publications indicate that exercise consistently increases mean salivary ?-amylase activities and concentrations, particularly at an intensity of >70% VO2max in healthy young individuals. Thus, these studies have confirmed that salivary ?-amylase levels markedly increase in response to physical stress. Salivary ?-amylase levels may therefore serve as an effective indicator in the non-invasive assessment of physical stress. PMID:24669232

  3. Enhancing Maritime Education and Training: Measuring a Ship Navigator's Stress Based on Salivary Amylase Activity

    ERIC Educational Resources Information Center

    Murai, Koji; Wakida, Shin-Ichi; Miyado, Takashi; Fukushi, Keiichi; Hayashi, Yuji; Stone, Laurie C.

    2009-01-01

    Purpose: The purpose of this paper is to propose that the measurement of salivary amylase activity is an effective index to evaluate the stress of a ship navigator for safe navigation training and education. Design/methodology/approach: Evaluation comes from the simulator and actual on-board experiments. The subjects are real captains who have…

  4. Caffeine administration does not alter salivary ?-amylase activity in young male daily caffeine consumers

    PubMed Central

    2014-01-01

    Background To follow up on a recent report from our lab [Hum Psychopharmacol 25:359–367, 2010.] we examined the effects of caffeine on salivary ?-amylase (sAA) activity in response to an engaging, non-stressful task in healthy young males (age 18–30 yrs) who consumed caffeine on a daily basis. Using a placebo-controlled, double-blind, between-subjects design, 45 men received either placebo, 200 mg or 400 mg of caffeine (Vivarin®). Participants then rested for 20 minutes, and performed a 20-minute computerized air traffic controller-like task that was cognitively engaging but not stressful. Saliva samples (assayed for sAA and cortisol), blood pressure, and heart rate were taken before (baseline) and 15 minutes after the computerized task. Results Systolic and diastolic blood pressure and sAA activity increased across the laboratory session (F’s?>?9.20, p’s?salivary cortisol levels decreased (F?=?16.17, p?salivary cortisol, or cardiovascular measures, and caffeine did not interact with the task to alter these measures. Conclusions Laboratory administered caffeine does not alter sAA activity, even when sAA activity is stimulated by participating in a cognitively engaging task. These data demonstrate that caffeine administration does not affect sAA activity, at least in healthy young men who regularly consume caffeine. Results support recent findings that basal caffeine levels in habitual caffeine users are not associated with basal sAA activity and that daily caffeine intake and diurnal sAA activity are not related. PMID:24410993

  5. April 16, 2010 Caffeine and Stress Alter Salivary -Amylase Activity in Young Men

    E-print Network

    Ritter, Frank

    , stress, blood pressure, heart rate *Address correspondence to: Laura Cousino Klein, Ph.D. Department examined the effects of caffeine and a psychological stressor on salivary -amylase (sAA) in healthy young (assayed for sAA and caffeine), blood pressure, and heart rate were taken before (baseline) and 15-mins

  6. Salivary alpha amylase activity in human beings of different age groups subjected to psychological stress.

    PubMed

    Sahu, Gopal K; Upadhyay, Seema; Panna, Shradha M

    2014-10-01

    Salivary alpha-amylase (sAA) has been proposed as a sensitive non-invasive biomarker for stress-induced changes in the body that reflect the activity of the sympathetic nervous system. Though several experiments have been conducted to determine the validity of this salivary component as a reliable stress marker in human subjects, the effect of stress induced changes on sAA level in different age groups is least studied. This article reports the activity of sAA in human subjects of different age groups subjected to psychological stress induced through stressful video clip. Differences in sAA level based on sex of different age groups under stress have also been studied. A total of 112 subjects consisting of both the male and female subjects, divided into two groups on basis of age were viewed a video clip of corneal transplant surgery as stressor. Activity of sAA from saliva samples of the stressed subjects were measured and compared with the activity of the samples collected from the subjects before viewing the clip. The age ranges of subjects were 18-25 and 40-60 years. The sAA level increased significantly in both the groups after viewing the stressful video. The increase was more pronounced in the younger subjects. The level of sAA was comparatively more in males than females in the respective groups. No significant change in sAA activity was observed after viewing the soothed video clip. Significant increase of sAA level in response to psychological stress suggests that it might act as a reliable sympathetic activity biochemical marker in different stages of human beings. PMID:25298630

  7. Salivary ?-amylase, serum albumin, and myoglobin protect against DNA-damaging activities of ingested dietary agents in vitro.

    PubMed

    Hossain, M Zulfiquer; Patel, Kalpesh; Kern, Scott E

    2014-08-01

    Potent DNA-damaging activities were seen in vitro from dietary chemicals found in coffee, tea, and liquid smoke. A survey of tea varieties confirmed genotoxic activity to be widespread. Constituent pyrogallol-like polyphenols (PLPs) such as epigallocatechin-3-gallate (EGCG), pyrogallol, and gallic acid were proposed as a major source of DNA-damaging activities, inducing DNA double-strand breaks in the p53R assay, a well characterized assay sensitive to DNA strand breaks, and comet assay. Paradoxically, their consumption does not lead to the kind of widespread cellular toxicity and acute disease that might be expected from genotoxic exposure. Existing physiological mechanisms could limit DNA damage from dietary injurants. Serum albumin and salivary ?-amylase are known to bind EGCG. Salivary ?-amylase, serum albumin, and myoglobin, but not salivary proline-rich proteins, reduced damage from tea, coffee, and PLPs, but did not inhibit damage from the chemotherapeutics etoposide and camptothecin. This represents a novel function for saliva in addition to its known functions including protection against tannins. Cell populations administered repeated pyrogallol exposures had abatement of measured DNA damage by two weeks, indicating an innate cellular adaptation. We suggest that layers of physiological protections may exist toward natural dietary products to which animals have had high-level exposure over evolution. PMID:24842839

  8. High Endogenous Salivary Amylase Activity Is Associated with Improved Glycemic Homeostasis following Starch Ingestion in Adults123

    PubMed Central

    Mandel, Abigail L.

    2012-01-01

    In the current study, we determined whether increased digestion of starch by high salivary amylase concentrations predicted postprandial blood glucose following starch ingestion. Healthy, nonobese individuals were prescreened for salivary amylase activity and classified as high (HA) or low amylase (LA) if their activity levels per minute fell 1 SD higher or lower than the group mean, respectively. Fasting HA (n = 7) and LA (n = 7) individuals participated in 2 sessions during which they ingested either a starch (experimental) or glucose solution (control) on separate days. Blood samples were collected before, during, and after the participants drank each solution. The samples were analyzed for plasma glucose and insulin concentrations as well as diploid AMY1 gene copy number. HA individuals had significantly more AMY1 gene copies within their genomes than did the LA individuals. We found that following starch ingestion, HA individuals had significantly lower postprandial blood glucose concentrations at 45, 60, and 75 min, as well as significantly lower AUC and peak blood glucose concentrations than the LA individuals. Plasma insulin concentrations in the HA group were significantly higher than baseline early in the testing session, whereas insulin concentrations in the LA group did not increase at this time. Following ingestion of the glucose solution, however, blood glucose and insulin concentrations did not differ between the groups. These observations are interpreted to suggest that HA individuals may be better adapted to ingest starches, whereas LA individuals may be at greater risk for insulin resistance and diabetes if chronically ingesting starch-rich diets. PMID:22492122

  9. Characterization of salivary alpha-amylase binding to Streptococcus sanguis.

    PubMed Central

    Scannapieco, F A; Bergey, E J; Reddy, M S; Levine, M J

    1989-01-01

    The purpose of this study was to identify the major salivary components which interact with oral bacteria and to determine the mechanism(s) responsible for their binding to the bacterial surface. Strains of Streptococcus sanguis, Streptococcus mitis, Streptococcus mutans, and Actinomyces viscosus were incubated for 2 h in freshly collected human submandibular-sublingual saliva (HSMSL) or parotid saliva (HPS), and bound salivary components were eluted with 2% sodium dodecyl sulfate. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western transfer, alpha-amylase (EC 3.2.1.1) was the prominent salivary component eluted from S. sanguis. Studies with 125I-labeled HSMSL or 125I-labeled HPS also demonstrated a component with an electrophoretic mobility identical to that of alpha-amylase which bound to S. sanguis. Purified alpha-amylase from human parotid saliva was radiolabeled and found to bind to strains of S. sanguis genotypes 1 and 3 and S. mitis genotype 2, but not to strains of other species of oral bacteria. Binding of [125I]alpha-amylase to streptococci was saturable, calcium independent, and inhibitable by excess unlabeled alpha-amylases from a variety of sources, but not by secretory immunoglobulin A and the proline-rich glycoprotein from HPS. Reduced and alkylated alpha-amylase lost enzymatic and bacterial binding activities. Binding was inhibited by incubation with maltotriose, maltooligosaccharides, limit dextrins, and starch. Images PMID:2788139

  10. Characterization of salivary alpha-amylase binding to Streptococcus sanguis

    SciTech Connect

    Scannapieco, F.A.; Bergey, E.J.; Reddy, M.S.; Levine, M.J. (State Univ. of New York, Buffalo (USA))

    1989-09-01

    The purpose of this study was to identify the major salivary components which interact with oral bacteria and to determine the mechanism(s) responsible for their binding to the bacterial surface. Strains of Streptococcus sanguis, Streptococcus mitis, Streptococcus mutans, and Actinomyces viscosus were incubated for 2 h in freshly collected human submandibular-sublingual saliva (HSMSL) or parotid saliva (HPS), and bound salivary components were eluted with 2% sodium dodecyl sulfate. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western transfer, alpha-amylase was the prominent salivary component eluted from S. sanguis. Studies with {sup 125}I-labeled HSMSL or {sup 125}I-labeled HPS also demonstrated a component with an electrophoretic mobility identical to that of alpha-amylase which bound to S. sanguis. Purified alpha-amylase from human parotid saliva was radiolabeled and found to bind to strains of S. sanguis genotypes 1 and 3 and S. mitis genotype 2, but not to strains of other species of oral bacteria. Binding of ({sup 125}I)alpha-amylase to streptococci was saturable, calcium independent, and inhibitable by excess unlabeled alpha-amylases from a variety of sources, but not by secretory immunoglobulin A and the proline-rich glycoprotein from HPS. Reduced and alkylated alpha-amylase lost enzymatic and bacterial binding activities. Binding was inhibited by incubation with maltotriose, maltooligosaccharides, limit dextrins, and starch.

  11. Salivary cortisol, salivary alpha amylase, and the dental anxiety scale.

    PubMed

    Sadi, Hana; Finkelman, Matthew; Rosenberg, Morton

    2013-01-01

    The aim of this study was to investigate the correlation between dental anxiety, salivary cortisol, and salivary alpha amylase (sAA) levels. Furthermore, the aim was to look into individual differences such as age, race, gender, any existing pain, or traumatic dental experience and their effect on dental anxiety. This study followed a cross-sectional design and included a convenience sample of 46. Every patient was asked to complete the Dental Anxiety Scale (DAS) and a basic demographic/dental history questionnaire. A saliva sample, utilizing the method of passive drooling, was then collected in 2-mL cryovials. Samples were analyzed for salivary cortisol and sAA levels by Salimetrics. Significant associations were observed between DAS scores and presence of pain and history of traumatic dental experience. However, no significant correlations were observed between DAS, cortisol, and sAA levels. Our study reconfirms that dental anxiety is associated with presence of pain and a history of traumatic dental experience. On the other hand, our study was the first to our knowledge to test the correlation between the DAS and sAA; nevertheless, our results failed to show any significant correlation between dental anxiety, cortisol, and sAA levels. PMID:23763559

  12. Salivary Cortisol, Salivary Alpha Amylase, and the Dental Anxiety Scale

    PubMed Central

    Sadi, Hana; Finkelman, Matthew; Rosenberg, Morton

    2013-01-01

    The aim of this study was to investigate the correlation between dental anxiety, salivary cortisol, and salivary alpha amylase (sAA) levels. Furthermore, the aim was to look into individual differences such as age, race, gender, any existing pain, or traumatic dental experience and their effect on dental anxiety. This study followed a cross-sectional design and included a convenience sample of 46. Every patient was asked to complete the Dental Anxiety Scale (DAS) and a basic demographic/dental history questionnaire. A saliva sample, utilizing the method of passive drooling, was then collected in 2-mL cryovials. Samples were analyzed for salivary cortisol and sAA levels by Salimetrics. Significant associations were observed between DAS scores and presence of pain and history of traumatic dental experience. However, no significant correlations were observed between DAS, cortisol, and sAA levels. Our study reconfirms that dental anxiety is associated with presence of pain and a history of traumatic dental experience. On the other hand, our study was the first to our knowledge to test the correlation between the DAS and sAA; nevertheless, our results failed to show any significant correlation between dental anxiety, cortisol, and sAA levels. PMID:23763559

  13. Anaerobic Threshold and Salivary ?-amylase during Incremental Exercise

    PubMed Central

    Akizuki, Kazunori; Yazaki, Syouichirou; Echizenya, Yuki; Ohashi, Yukari

    2014-01-01

    [Purpose] The purpose of this study was to clarify the validity of salivary ?-amylase as a method of quickly estimating anaerobic threshold and to establish the relationship between salivary ?-amylase and double-product breakpoint in order to create a way to adjust exercise intensity to a safe and effective range. [Subjects and Methods] Eleven healthy young adults performed an incremental exercise test using a cycle ergometer. During the incremental exercise test, oxygen consumption, carbon dioxide production, and ventilatory equivalent were measured using a breath-by-breath gas analyzer. Systolic blood pressure and heart rate were measured to calculate the double product, from which double-product breakpoint was determined. Salivary ?-amylase was measured to calculate the salivary threshold. [Results] One-way ANOVA revealed no significant differences among workloads at the anaerobic threshold, double-product breakpoint, and salivary threshold. Significant correlations were found between anaerobic threshold and salivary threshold and between anaerobic threshold and double-product breakpoint. [Conclusion] As a method for estimating anaerobic threshold, salivary threshold was as good as or better than determination of double-product breakpoint because the correlation between anaerobic threshold and salivary threshold was higher than the correlation between anaerobic threshold and double-product breakpoint. Therefore, salivary threshold is a useful index of anaerobic threshold during an incremental workload. PMID:25140097

  14. Usefulness of salivary alpha amylase as a biomarker of chronic stress and stress related oral mucosal changes – a pilot study

    PubMed Central

    Pai, Keerthilatha-M.; Vengal, Manoj; Gopalakrishna, Kodyalamoole; Narayanakurup, Dinesh

    2014-01-01

    Introduction: Salivary biomarkers are suggested to provide a reliable, noninvasive and objective measurement of chronic psychosocial stress and helps in assessment of pivotal role of stress in causation or precipitation of multitude of health problems. Objectives: To evaluate the usefulness of salivary alpha amylase activity as an objective indicator of chronic stress and to find out any correlation between stress- related mucosal complaints and its levels. Study Design: Study was conducted among 50 subjects suffering from chronic stress related problems and 50 non-stressed individuals who were screened with a psychometric questionnaire. Brief case history and oral examination was carried out and about one ml of unstimulated saliva was collected. Salivary alpha amylase levels estimated were compared between study and control group and between subjects with and without oral mucosal changes using non parametric Mann Whitney U test. Results: There was statistically significant higher salivary alpha amylase levels in study group (p =.002) and salivary alpha amylase between the oral mucosal complaints group and without oral mucosal complaints group within the total study population were found to be statistically significant (p=0.045). Conclusions: Salivary amylase activity increases in patients with chronic psychosocial stress and may be used as a biomarker of chronic stress, but it may not be an indicator to suggest the development of stress related oral mucosal changes. Key words:Salivary biomarker, salivary alpha amylase, psychosocial stress, sympathetic nervous system, oral mucosal changes. PMID:24790712

  15. Salivary ?-amylase levels as a biomarker of experienced fear

    PubMed Central

    Bibas, David; Adolphs, Ralph

    2010-01-01

    We recently reported data related to emotions collected in conjunction with a museum exhibit on emotion (Goose Bumps!—The Science of Fear).1 In this addendum, we present additional data collected as part of that study. We collected two commonly measured indices of emotional arousal, salivary cortisol and ?-amylase, before and after participants had gone through a realistic fear challenge course as part of the exhibit. We found that ?-amylase, but not cortisol, showed a highly specific increase only for those participants who endorsed both emotional arousal and negative valence. By contrast, the fear-inducing course resulted in high arousal but positive valence in some participants; in these, no increased ?-amylase was measured. We conclude that salivary ?-amylase is a promising biomarker for fearful experiences, and suggest that it is important to pay attention to positively valenced arousal that may be induced by fearful stimuli in a laboratory setting. PMID:21331229

  16. Interparental Aggression and Parent-Adolescent Salivary Alpha Amylase Symmetry

    PubMed Central

    Gordis, Elana B.; Margolin, Gayla; Spies, Lauren; Susman, Elizabeth J.; Granger, Douglas A.

    2010-01-01

    The present study examined salivary alpha-amylase (sAA), a putative marker of adrenergic activity, in family members engaging in family conflict discussions. We examined symmetry among family members' sAA levels at baseline and in response to a conflict discussion. The relation between a history of interparental aggression on parent-adolescent sAA symmetry also was examined. Participants were 62 families with a mother, father, and biological child age 13-18 (n = 29 girls). After engaging in a relaxation procedure, families participated in a 15-minute triadic family conflict discussion. Participants provided saliva samples at post-relaxation/pre-discussion, immediately post-discussion, and at 10 and 20 min post-discussion. Participants also reported on interparental physical aggression during the previous year. Across the sample we found evidence of symmetry between mothers' and adolescents' sAA levels at baseline and around the discussion. Interparental aggression was associated with lower sAA levels among fathers. Interparental aggression also affected patterns of parent-child sAA response symmetry such that families reporting interparental aggression exhibited greater father-adolescent sAA symmetry than did those with no reports of interparental aggression. Among families with no interparental aggression history, we found consistent mother-adolescent symmetry. These differences suggest different patterns of parent-adolescent physiological attunement among families with interparental aggression. PMID:20096715

  17. Multiple time courses of salivary alpha-amylase and dimensions of affect in adolescence.

    PubMed

    Doane, Leah D; Van Lenten, Scott A

    2014-11-01

    Previous research has illustrated associations among daily experiences, emotions and stress-responding physiological systems. Recently, investigators have examined salivary alpha-amylase (sAA), a surrogate marker of the autonomic nervous system, and its associations with affect. The current study examined associations among affective valence, arousal and sAA across three different time courses at the momentary, daily and inter-individual level to understand varying influences of adolescents' daily emotional experiences on sAA reactivity and diurnal sAA activity. Adolescents (N=82) provided salivary samples and diary reports of affect and experiences five times a day for three consecutive days. They also completed self-report questionnaires on trait affect. Findings from multilevel growth curves demonstrated that adolescents in our sample displayed typical sAA diurnal rhythms with levels dropping 30 min after waking and then increasing across the day to a peak in the late afternoon. Within person momentary experiences of high arousal positive affect were associated with momentary sAA reactivity. Prior day experiences of high arousal negative affect were associated with a greater amylase awakening response (i.e., greater decrease) and flatter slopes the next day. Trait positive affect was also associated with flatter sAA slopes. Our findings suggest that both affective arousal and valence should be accounted for when examining differences in sAA reactivity and diurnal patterns. Further, our results indicated that emotion-physiology transactions among adolescents occur over varying time scales for salivary alpha-amylase as well as cortisol. PMID:25076484

  18. The psychosocial stress-induced increase in salivary alpha-amylase is independent of saliva flow rate

    Microsoft Academic Search

    Nicolas Rohleder; Jutta M. Wolf; Enrique F. Maldonado; Clemens Kirschbaum

    2006-01-01

    The stress response of salivary alpha-amylase (sAA) has been suggested as an index for sympathetic nervous system activation. However, concurrent inhibition of the parasympathetic nervous system is discussed as a confounder due to suppression of saliva flow rate. Here we set out to test the influence of stress-induced changes in flow rate on sAA secretion. Twenty-six subjects underwent the Trier

  19. Discovering an Accessible Enzyme: Salivary [alpha]-Amylase--"Prima Digestio Fit in Ore"--A Didactic Approach for High School Students

    ERIC Educational Resources Information Center

    Marini, Isabella

    2005-01-01

    Human salivary [alpha]-amylase is used in this experimental approach to introduce biology high school students to the concept of enzyme activity in a dynamic way. Through a series of five easy, rapid, and inexpensive laboratory experiments students learn what the activity of an enzyme consists of: first in a qualitative then in a semi-quantitative…

  20. Daytime Secretion of Salivary Cortisol and Alpha-Amylase in Preschool-Aged Children with Autism and Typically Developing Children

    ERIC Educational Resources Information Center

    Kidd, Sharon A.; Corbett, Blythe A.; Granger, Douglas A.; Boyce, W. Thomas; Anders, Thomas F.; Tager, Ira B.

    2012-01-01

    We examined daytime salivary cortisol and salivary alpha-amylase (sAA) secretion levels and variability in preschool-aged children with autism (AUT) and typically developing children (TYP). Fifty-two subjects (26 AUT and 26 TYP) were enrolled. Salivary samples were obtained at waking, midday, and bedtime on two consecutive days at three phases…

  1. Stability of human ?-salivary amylase in aged forensic samples.

    PubMed

    Carboni, Ilaria; Rapi, Stefano; Ricci, Ugo

    2014-07-01

    The unequivocal tissue identification in forensic casework samples is a key step for crime scene reconstruction. Just knowing the origin of a fluid can sometimes be enough to either prove or disprove a fact in court. Despite the importance of this test, very few data are available in literature concerning human saliva identification in old forensic caseworks. In this work the stability of human ?-amylase activity in aged samples is described by using three different methods integrated with DNA profiling techniques. This analytical protocol was successfully applied on 26-years old samples coming from anonymous threat letters sent to prosecutors who were working on "the Monster of Florence", a case of serial murders happened around Florence (Italy) between 1968 and 1985. PMID:24755314

  2. Peer Victimization and Aggression: Moderation by Individual Differences in Salivary Cortisol and Alpha-Amylase

    ERIC Educational Resources Information Center

    Rudolph, Karen D.; Troop-Gordon, Wendy; Granger, Douglas A.

    2010-01-01

    This research examined whether variations in salivary measures of the hypothalamic-pituitary-adrenal axis (cortisol) and autonomic nervous system (alpha amylase [sAA]) contribute to individual differences in the association between peer victimization and aggression. Children (N = 132; M age = 9.46 years, SD = 0.33) completed a measure of peer…

  3. Estimation of Levels of Salivary Mucin, Amylase and Total Protein in Gingivitis and Chronic Periodontitis Patients

    PubMed Central

    Bhandary, Rahul; Thomas, Biju; Kumari, Suchetha

    2014-01-01

    Background: Periodontal diseases are a group of inflammatory conditions resulting from interaction between a pathogenic bacterial biofilm and susceptible host’s inflammatory response eventually leading to the destruction of periodontal structures and subsequent tooth loss. Hence, investigation of salivary proteins in individuals with periodontal diseases may be useful to enhance the knowledge of their roles in these diseases. Materials and Methods: This case-control study was conducted at A.B. Shetty Memorial Institute of Dental Sciences, Mangalore. The study comprised of 90 patients of age between 25-60 years who were clinically examined and divided into three groups of 30 each: namely clinically healthy, gingivitis and chronic periodontitis. These were classified according to the values of gingival index score, clinical attachment loss and probing pocket depth. Unstimulated saliva was collected and salivary mucin, amylase and total protein levels were determined. Statistical analysis: Results obtained were tabulated and statistically analyzed using ANOVA test and Karl pearson’s correlation test. Results: The results of the study showed an increased concentration of salivary mucin, amylase and total protein in gingivitis patients and increased levels of amylase and total protein in saliva of chronic periodontitis patients compared to healthy individuals which were statistically significant. A decrease in mucin concentration was observed in the periodontitis group compared to gingivitis group. A positive correlation was present between salivary mucin, amylase and total protein levels in the three groups. Conclusion: Salivary mucin, amylase and total protein may serve as an important biochemical parameter of inflammation of the periodontium. Also, it can be hypothesized that various enzyme inhibitors might be useful as a part of host modulation therapy in the treatment of periodontal diseases. PMID:25478449

  4. Correlation of Salivary Alpha Amylase Level and Adenotonsillar Hypertrophy with Sleep Disordered Breathing in Pediatric Subjects

    PubMed Central

    Park, Chan-Soon; Guilleminault, Christian; Park, Hong-Jin; Cho, Jin-Hee; Lee, Heung-Ku; Son, Hye-Lim; Hwang, Se-Hwan

    2014-01-01

    Study Objectives: Obstructive sleep apnea syndrome (OSAS) and sleep disordered breathing (SDB) can affect the sympathetic adrenomedullary system (SAM). As a biomarker of SAM activity, salivary ?-amylase (sAA) in pediatric subjects was evaluated whether it has any correlation with polysomnographic (PSG) parameters related to SDB. Methods: Sixty-seven children who attended our clinic during 1 year were enrolled prospectively and underwent clinical examinations and in-lab polysomnography. The sAA was measured at 2 points—at night before PSG and in the early morning after PSG Results: Subjects were divided into control (n = 26, apneahypopnea index [AHI] < 1) and OSAS (n = 41, AHI ? 1) groups. The OSAS group was subdivided according to AHI (mild-moderate, 1 ? AHI < 10; severe, AHI ? 10). The sAA subtraction and ratio (p = 0.014 and p < 0.001, respectively) were significantly higher in severe OSAS than in the mild-moderate and control groups. Although oxygen desaturation index (ODI) and AHI were significantly associated with sAA, sAA in the OSAS group was not related to lowest oxygen saturation or adenotonsillar hypertrophy. Conclusion: sAA was well related to polysomnographic (PSG) parameters related to SDB, such as AHI and ODI. Therefore, screening test for sAA in children suspected to have SBD may help to identify OSAS patients from control. Citation: Park CS, Guilleminault C, Park HJ, Cho JH, Lee HK, Son HL, Hwang SH. Correlation of salivary alpha amylase level and adenotonsillar hypertrophy with sleep disordered breathing in pediatric subjects. J Clin Sleep Med 2014;10(5):559-566. PMID:24812542

  5. Salivary Alpha Amylase and Cortisol Levels in Children with Global Developmental Delay and Their Relation with the Expectation of Dental Care and Behavior during the Intervention

    ERIC Educational Resources Information Center

    dos Santos, Marcio Jose Possari; Bernabe, Daniel Galera; Nakamune, Ana Claudia de Melo Stevanato; Perri, Silvia Helena Venturoli; de Aguiar, Sandra Maria Herondina Coelho Avila; de Oliveira, Sandra Helena Penha

    2012-01-01

    The purpose of this study was to analyze the alpha-amylase (sAA) and cortisol levels in children with Global developmental delay (GDD) before and after dental treatment and its association with the children's behavior during treatment. The morning salivary cortisol levels and activity of sAA of 33 children with GDD were evaluated before and after…

  6. Diurnal patterns of salivary alpha-amylase and cortisol secretion in female adolescent tennis players after 16 weeks of training.

    PubMed

    Filaire, Edith; Ferreira, Jose Pedro; Oliveira, Miguel; Massart, Alain

    2013-07-01

    We examined the effects of 16 weeks of training on diurnal pattern of salivary alpha-amylase (sAA), cortisol, and the ratio of sAA over cortisol (AOC) in 12 national adolescent female tennis players. Stress and recovery were also evaluated using the Recovery-Stress-Questionnaire for Athletes-RESTQ-Sport. Data were collected after a 2-week rest (January, W0), and 4 months after W0 (W16). Subjects collected five saliva samples throughout a day. While all participants displayed the previously shown decrease after awakening in adolescents at W0, they showed a rise in the alpha-amylase awakening response and a higher alpha-amylase activity output (p<0.01) at W16 compared to W0. For the daily rhythm of cortisol we found subjects having a low overall output of salivary cortisol (p<0.01) and a blunted response to awakening at W16. Furthermore, an increase in the ratio AOC at W16, and a negative correlation between this ratio and Sport-specific recovery score. Our findings offer support for the hypothesis that increase of training load during the study period induced asymmetry activation between the two stress systems, in relation to psychological alterations and performance decrease. These results provide encouragement to continue exploring the impact of training program using a psychobiological approach among young athletes in order to prevent fatigue and preserve the health of these athletes. PMID:23200107

  7. Salivary cortisol and alpha-amylase reactivity to taekwondo competition in children

    Microsoft Academic Search

    Corrado Lupo; Cristina Cortis; Salvatore Chiodo; Giuseppe Cibelli; Antonio Tessitore

    The aim of this study was to evaluate the effects of an official taekwondo competition (three 1-min rounds with a 1-min recovery\\u000a in-between) on heart rate (HR), salivary alpha-amylase (sAA), and salivary-free cortisol (sC) in children. Parental consent\\u000a was obtained for 12 young (10.4 ± 0.2 years) male taekwondo athletes. Saliva sample were collected 15 min before and 1 min\\u000a after an official taekwondo competition,

  8. Activity of alpha-amylase inhibitors from Phaseolus coccineus on digestive alpha-amylases of the coffee berry borer.

    PubMed

    Valencia-Jiménez, Arnubio; Arboleda Valencia, Jorge W; Grossi-De-Sá, Maria Fátima

    2008-04-01

    Seeds of scarlet runner bean ( Phaseolus coccineus L.) were analyzed for alpha-amylase inhibitor (alpha-AI) activity. Through the use of polyclonal antibodies raised against pure alpha-AI-1 from common bean ( Phaseolus vulgaris L.), typical alpha-AlphaIota polypeptides (Mr 14-18 kDa) as well as a large polypeptide of Mr 32000 Da, usually referred to as "amylase inhibitor like", were detected. The inhibitor activity present in four accessions of P. coccineus was examined, both in semiquantitative zymograms allowing the separation of different isoforms and in quantitative assays against human salivary amylase, porcine pancreatic amylase, and coffee berry borer, Hypothenemus hampei Ferrari (Coleoptera: Scolytidae) amylase. Differential inhibition curves lead to the suggestion that the gene encoding one of the inhibitors in P. coccineus (in accession G35590) would be a good candidate for genetic engineering of coffee resistance toward the coffee berry borer. An in vitro proteolytic digestion treatment of pure alpha-AlphaIota-1 resulted in a rapid loss of the inhibitory activity, seriously affecting its natural capacity to interact with mammalian alpha-amylases. PMID:18321052

  9. Alpha amylase as a salivary biomarker of acute stress of venepuncture from periodic medical examinations.

    PubMed

    Koh, David; Ng, Vivian; Naing, Lin

    2014-01-01

    Periodic occupational health examinations often require venepuncture. Acute psychological and physical stressors during such procedure result in sympathetic stimulation and increased salivary protein secretion, including salivary ?-amylase (SAA). We studied SAA response to venepuncture during such examination. Fifty-eight healthy males undergoing periodic medical examination reported perceived stress level (PSL) scores (on a five-point scale) and provided passive drool saliva samples at 15-min (T1) and 1-min before (T2); and 1-min (T3) and 15-min after venepuncture (T4). A subset of 33 participants available for repeat examination on a control day when there was no venepuncture provided saliva samples at the corresponding times for comparison. Saliva SAA activity levels were analyzed using a SAA assay kit (Salimetrics LLC, USA). Among 58 participants, mean SAA increased from T1 (89.95?U/L) to T2 (109.5?U/L) and T3 (116.9?U/L). SAA remained elevated 15?min after venepuncture (121.0?U/L). A positive trend in the difference of SAA between T3 and T1 was noted among subjects with increasing mean PSL scores. T3-T1 values were 0.6 (among those with PSL???1, n?=?24), 11.3 (among those with PSL between 1 and 1.5, n?=?18), and 78.9 (among those with PSL?>?1.5, n?=?16). SAA increment over four-time points was significantly higher on the venepuncture compared to the control day (P?=?0.021). SAA increases in response to the acute stress of venepuncture during a periodic medical examination, and remains elevated 15?min after the procedure. In comparison, such fluctuations in SAA were not seen on a control day. During venepuncture, increase in SAA from baseline is higher among those who reported greater self-perceived stress during the procedure. PMID:25207265

  10. Alpha Amylase as a Salivary Biomarker of Acute Stress of Venepuncture from Periodic Medical Examinations

    PubMed Central

    Koh, David; Ng, Vivian; Naing, Lin

    2014-01-01

    Periodic occupational health examinations often require venepuncture. Acute psychological and physical stressors during such procedure result in sympathetic stimulation and increased salivary protein secretion, including salivary ?-amylase (SAA). We studied SAA response to venepuncture during such examination. Fifty-eight healthy males undergoing periodic medical examination reported perceived stress level (PSL) scores (on a five-point scale) and provided passive drool saliva samples at 15-min (T1) and 1-min before (T2); and 1-min (T3) and 15-min after venepuncture (T4). A subset of 33 participants available for repeat examination on a control day when there was no venepuncture provided saliva samples at the corresponding times for comparison. Saliva SAA activity levels were analyzed using a SAA assay kit (Salimetrics LLC, USA). Among 58 participants, mean SAA increased from T1 (89.95?U/L) to T2 (109.5?U/L) and T3 (116.9?U/L). SAA remained elevated 15?min after venepuncture (121.0?U/L). A positive trend in the difference of SAA between T3 and T1 was noted among subjects with increasing mean PSL scores. T3–T1 values were 0.6 (among those with PSL???1, n?=?24), 11.3 (among those with PSL between 1 and 1.5, n?=?18), and 78.9 (among those with PSL?>?1.5, n?=?16). SAA increment over four-time points was significantly higher on the venepuncture compared to the control day (P?=?0.021). SAA increases in response to the acute stress of venepuncture during a periodic medical examination, and remains elevated 15?min after the procedure. In comparison, such fluctuations in SAA were not seen on a control day. During venepuncture, increase in SAA from baseline is higher among those who reported greater self-perceived stress during the procedure. PMID:25207265

  11. Estimation of Salivary Amylase in Diabetic Patients and Saliva as a Diagnostic Tool in Early Diabetic Patients

    PubMed Central

    Malathi, L.; Masthan, K.M.K.; Balachander, N.; Babu, N. Aravindha; Rajesh, E.

    2013-01-01

    Aim: The aim of this study was to estimate the salivary amylase levels in non-insulin dependent diabetes mellitus patients and to correlate these findings with those in normal individuals, in order to provide salivary amylase level as a bio-chemical indicator for diagnosing and monitoring the glucose levels. Material and Methods: The study samples consisted of 60 individuals. Both males and females participated in the study. Thirty non-insulin dependent diabetes mellitus patients of age group of 30 to 60 years and healthy individuals of same number and age group were included in this study. The data obtained in this study were statistically analyzed by using Student’s t–test. Results: In estimation of salivary amylase levels, the comparison of mean and standard deviation showed the highest mean score (2739.48 +1525.20) among the diabetic patients and lowest mean score (1740.38 + 638.51) among the non-diabetic patients. The p-value obtained was less than 0.01. Hence, a highly significant difference in the mean scores regarding salivary amylase (u/l) was found among the two groups. Conclusion: The mean scores of age, fasting blood sugar, post prandial blood sugar, HbA1c and salivary amylase levels were greater in diabetic patients than in non-diabetic patients. PMID:24392426

  12. Sociodemographic Risk, Parenting, and Effortful Control: Relations to Salivary Alpha-amylase and Cortisol in Early Childhood

    PubMed Central

    Taylor, Zoe E.; Spinrad, Tracy L.; VanSchyndel, Sarah K.; Eisenberg, Nancy; Huynh, Jacqueline; Sulik, Michael J.; Granger, Douglas A.

    2012-01-01

    Early sociodemographic risk, parenting, and temperament were examined as predictors of the activity of children’s (N = 148; 81 boys, 67 girls) hypothalamic-pituitary-adrenal axis and autonomic nervous system. Demographic risk was assessed at 18 months (T1), intrusive-overcontrolling parenting and effortful control were assessed at 30 months (T2), and salivary cortisol and alpha-amylase were collected at 72 (T3) months of age. Demographic risk at T1 predicted lower levels of children’s effortful control and higher levels of mothers’ intrusive-overcontrolling parenting at T2. Intrusive-overcontrolling parenting at T2 predicted higher levels of children’s cortisol and alpha-amylase at T3, but effortful control did not uniquely predict children’s cortisol or alpha-amylase. Findings support the open nature of stress responsive physiological systems to influence by features of the early caregiving environment and underscore the utility of including measures of these systems in prevention trials designed to influence child outcomes by modifying parenting behavior. PMID:22949301

  13. Sex Differences in Salivary Cortisol, Alpha-Amylase, and Psychological Functioning Following Hurricane Katrina

    ERIC Educational Resources Information Center

    Vigil, Jacob M.; Geary, David C.; Granger, Douglas A.; Flinn, Mark V.

    2010-01-01

    The study examines group and individual differences in psychological functioning and hypothalamic-pituitary-adrenal and sympathetic nervous system (SNS) activity among adolescents displaced by Hurricane Katrina and living in a U.S. government relocation camp (n = 62, ages 12-19 years) 2 months postdisaster. Levels of salivary cortisol, salivary

  14. Collecting saliva and measuring salivary cortisol and alpha-amylase in frail community residing older adults via family caregivers.

    PubMed

    Hodgson, Nancy A; Granger, Douglas A

    2013-01-01

    Salivary measures have emerged in bio-behavioral research that are easy-to-collect, minimally invasive, and relatively inexpensive biologic markers of stress. This article we present the steps for collection and analysis of two salivary assays in research with frail, community residing older adults-salivary cortisol and salivary alpha amylase. The field of salivary bioscience is rapidly advancing and the purpose of this presentation is to provide an update on the developments for investigators interested in integrating these measures into research on aging. Strategies are presented for instructing family caregivers in collecting saliva in the home, and for conducting laboratory analyses of salivary analytes that have demonstrated feasibility, high compliance, and yield quality specimens. The protocol for sample collection includes: (1) consistent use of collection materials; (2) standardized methods that promote adherence and minimize subject burden; and (3) procedures for controlling certain confounding agents. We also provide strategies for laboratory analyses include: (1) saliva handling and processing; (2) salivary cortisol and salivary alpha amylase assay procedures; and (3) analytic considerations. PMID:24378361

  15. Attenuated acute salivary ?-amylase responses to gustatory stimulation with citric acid in thin children.

    PubMed

    Chen, Long Hui; Yang, Ze Min; Chen, Wei Wen; Lin, Jing; Zhang, Min; Yang, Xiao Rong; Zhao, Ling Bo

    2015-04-01

    Salivary ?-amylase (sAA) is responsible for the 'pre-digestion' of starch in the oral cavity and accounts for up to 50 % of salivary protein in human saliva. An accumulating body of literature suggests that sAA is of nutritional importance; however, it is still not clear how sAA is related to individual's nutritional status. Although copy number variations (CNV) of the salivary amylase gene (AMY1) are associated with variation in sAA levels, a significant amount of sAA variation is not explained by AMY1 CNV. To measure sAA responses to gustatory stimulation with citric acid, we used sAA ratio (the ratio of stimulated sAA levels to those of resting sAA) and investigated acute sAA responses to citric acid in children with normal (Normal-BMI, n 22) and low (Low-BMI, n 21) BMI. The AMY1 gene copy number was determined by quantitative PCR. We, for the first time, demonstrated attenuated acute sAA responses (decreased sAA ratio) to gustatory stimulation in Low-BMI (thinness grade 3) children compared with the Normal-BMI children, which suggest that sAA responses to gustatory stimulation may be of nutritional importance. However, child's nutritional status was not directly related to their resting or stimulated sAA levels, and it was not associated with AMY1 gene copy number. Finally, AMY1 CNV might influence, but did not eventually determine, sAA levels in children. PMID:25784372

  16. Reduction in Salivary ?-amylase Levels following a Mind-Body Intervention in Cancer Survivors - an Exploratory Study

    PubMed Central

    Lipschitz, David L.; Kuhn, Renee; Kinney, Anita Y.; Donaldson, Gary W.; Nakamura, Yoshio

    2013-01-01

    Objective The main aim of this exploratory study was to assess whether salivary ?-amylase (sAA) and salivary cortisol levels would be positively modulated by sleep-focused mind-body interventions in female and male cancer survivors. Methods We conducted a randomized controlled trial in which 57 cancer survivors with self-reported sleep disturbance received either a Sleep Hygiene Education (SHE; n=18) control, or one of two experimental mind-body interventions, namely, Mind-Body Bridging (MBB; n=19) or Mindfulness Meditation (MM; n=20). Interventions were three sessions each conducted once per week for three consecutive weeks. Saliva cortisol and sAA were measured at baseline and one week after the last session. Participants also completed a sleep scale at the same time points when saliva was collected for biomarker measurement. Results Our study revealed that at post-study assessment, mean sAA levels upon awakening (“Waking” sample) declined in MBB compared with that of SHE. Mean Waking cortisol levels did not differ among treatment groups but declined slightly in SHE. Self-reported sleep improved across the three interventions at Post-assessment, with largest improvements in the MBB intervention. Conclusion In this exploratory study, sleep focused mind-body intervention (MBB) attenuated Waking sAA levels, suggesting positive influences of a mind-body intervention on sympathetic activity in cancer survivors with sleep disturbance. PMID:23375640

  17. Salivary Alpha Amylase as a Noninvasive Biomarker for Dental Fear and Its Correlation with Behavior of Children during Dental Treatment

    PubMed Central

    Noorani, Hina; Shivaprakash, PK

    2014-01-01

    ABSTrACT Objective: Objectives of our studies were to predict dental fear in a child patient depending on salivary alpha amylase (sAA) level before and after dental treatment and to evaluate correla­tion of later with behavior of child patient during dental treatment. Materials and methods: Seventy-seven children between age of 5 and 12 years were divided in three groups. Group 1 consisted of 25 school children who did not undergo any dental treatment. Groups 2 and 3 underwent dental treatment without and with local anesthesia respectively. Groups 2 and 3 were administered child fear survey schedule-dental subscale (CFSS-DS) questionnaire before treatment. Salivary samples were collected for sAA estimation in groups 2 and 3 children before and after completion of dental treatment and behavior during treatment was noted using Frankel behavior rating scale. Group 1 acted as control in which salivary sample was collected in absence of dental stress. Results: When groups 2 and 3 were combined, pretreatment sAA level had a statistically significant (p = 0.0094) correlation with CFSS-DS scores. Conclusion: Alpha amylase can be used as a screening tool to predict level of dental fear in a child patient. How to cite this article: Noorani H, Joshi HV, Shivaprakash PK. Salivary Alpha Amylase as a Noninvasive Biomarker for Dental Fear and Its Correlation with Behavior of Children during Dental Treatment. Int J Clin Pediatr Dent 2014;7(1):19-23. PMID:25206232

  18. Effect of lecturing to 200 students on heart rate variability and alpha-amylase activity

    Microsoft Academic Search

    Edith Filaire; Hugues Portier; Alain Massart; Luis Ramat; Anna Teixeira

    2010-01-01

    The aim of this study was to examine cardiovascular [heart rate variability (HRV)] and autonomic nervous system activation\\u000a (by evaluating salivary alpha-amylase activity) that occur in professors both to, and after, the delivery of a lecture to\\u000a 200 students and to determine whether gender is an influencing factor upon response. Fifty-two participants (26 women and\\u000a 26 men) collected eight unstimulated

  19. Individual Differences in Preschoolers' Salivary Cortisol and Alpha-Amylase Reactivity: Relations to Temperament and Maladjustment

    PubMed Central

    Spinrad, Tracy L.; Eisenberg, Nancy; Granger, Douglas A.; Eggum, Natalie D.; Sallquist, Julie; Haugen, RG; Kupfer, Anne; Hofer, Claire

    2009-01-01

    We examined the relations of 84 preschoolers' (43 boys; mean age = 54 months) situational stress reactivity to their observed emotions and mothers' reports of temperament and adjustment. Salivary cortisol and salivary alpha-amylase (sAA) were collected prior to, and following, a frustrating task. Children's anger, sadness, and positive affect were measured, and mothers reported on preschoolers' dispositional emotionality, regulation, impulsivity, and problem behaviors. Forty-seven percent of children had an increase in sAA and 52% had an increase in cortisol following the challenging task. On average, sAA levels showed the predicted pattern of rise following the frustrating task, followed by return to baseline. For cortisol, there was a mean increase from pre-task to 40 minutes post-test. sAA reactivity was associated with relatively low levels of dispositional anger and impulsivity and relatively high regulation, particularly for girls. sAA reactivity also was related to low externalizing problems for girls, but not boys. Although cortisol reactivity was unrelated to children's emotions and maladjustment, it was positively related to mothers' reports of regulation. The findings suggest that sAA reactivity in response to a frustrating social task may reflect girls' constrained behavior. PMID:19348808

  20. An in vitro and in vivo study of the ?-amylase activity of phaseolamin.

    PubMed

    de Gouveia, Neire Moura; Alves, Fernanda Vieira; Furtado, Fabiana Barcelos; Scherer, Danielli Luana; Mundim, Antonio Vicente; Espindola, Foued Salmen

    2014-08-01

    We evaluated the polypeptide profiles, inhibition of human salivary ?-amylase activity, and hemagglutination properties of a commercial phaseolamin sample. We also performed an in vivo assay to investigate the effects of a commercial phaseolamin treatment (100, 500, or 1500?mg/kg) over 20 days on the glycemia, body weight, and serum biochemical parameters (total cholesterol, triglycerides, alanine aminotransferase, and aspartate aminotransferase) of nondiabetic and streptozotocin-induced diabetic rats. The in vitro evaluation showed defined protein profiles, low hemagglutination activity, and high ?-amylase inhibition. None of the experimental groups treated with phaseolamin or acarbose showed decreases in body weight. Our data demonstrate that phaseolamin inhibits amylase activity in vitro, reduces blood glucose levels, decreases or attenuates some of the renal and hepatic effects of diabetes in streptozotocin-induced rats, and could therefore have therapeutic potential in the treatment or prevention of the complications of diabetes. PMID:24650210

  1. Structure of human salivary ?-amylase crystallized in a C-centered monoclinic space group

    PubMed Central

    Fisher, S. Zoë; Govindasamy, Lakshmanan; Tu, Chingkuang; Agbandje-McKenna, Mavis; Silverman, David N.; Rajaniemi, Hannu J.; McKenna, Robert

    2006-01-01

    Human salivary ?-amylase (HSA) is a major secretory protein component of saliva and has important biological functions, including the initial digestion of starch. HSA acts as a monomer and mediates the hydrolysis of ?-1,4-glucosidic linkages in oligosaccharides. To date, all published crystal structures of HSA have been crystallized as monomers in space group P212121. Here, the serendipitous purification, crystallization and ultimate structure determination of a HSA non-crystallographic symmetry (NCS) dimer, while attempting to purify human carbonic anhydrase VI (HCA VI) from saliva using an affinity resin for ?-class carbonic anhydrases, is presented. On further investigation, it was shown that HSA could only be copurified using the affinity resin in the presence of HCA VI which is glycosylated and not the non-glycosylated HCA II. The identification of the HSA crystals was carried out by peptide mapping and mass spectrometry. HSA was shown to have crystallized as an NCS dimer in space group C2, with unit-cell parameters a = 150.9, b = 72.3, c = 91.3?Å, ? = 102.8°. The NCS dimer crystal structure is reported to 3.0?Å resolution, with a refined R cryst of 0.228. The structure is compared with the previously reported P212121 monomer structures and the crystal packing and dimer interface are discussed. PMID:16511271

  2. Salivary cortisol and alpha-amylase reactivity to taekwondo competition in children.

    PubMed

    Capranica, Laura; Lupo, Corrado; Cortis, Cristina; Chiodo, Salvatore; Cibelli, Giuseppe; Tessitore, Antonio

    2012-02-01

    The aim of this study was to evaluate the effects of an official taekwondo competition (three 1-min rounds with a 1-min recovery in-between) on heart rate (HR), salivary alpha-amylase (sAA), and salivary-free cortisol (sC) in children. Parental consent was obtained for 12 young (10.4 ± 0.2 years) male taekwondo athletes. Saliva sample were collected 15 min before and 1 min after an official taekwondo competition, and at 30, 60, and 90 min of the recovery period. To evaluate the exercise intensity during the competition, HR was measured and expressed as a percentage of individuals HR(peak). Athletes spent 78% of the time working at HR > 90% HR(max), with significant increases from round 1 to round 2 and 3. Peak sAA observed at the end of the match (169.6 ± 47.0 U/mL) was different (P = 0.0001) from the other samplings (pre-competition 55.0 ± 14.0 U/mL, 30-min recovery 80.4 ± 17.7 U/mL, 60-min recovery 50.5 ± 7.6 U/ml; 90-min recovery 53.2 ± 9.6 U/mL). Peak sC values observed at 30-min recovery (17.9 ± 3.5 nmol/L) were different (P < 0.0001) from pre-competition (5.6 ± 0.9 nmol/L), post-competition (9.0 ± 2.0 nmol/L), 60-min recovery (10.3 ± 2.6 nmol/L) and 90-min recovery (4.2 ± 0.8 nmol/L) values. These findings confirm that taekwondo competitions pose a high stress on young athletes. The different sAA and sC reactions in response to the physical stressor mirror the faster reactivity of the sympathetic-adrenomedullary system relatively to the hypothalamic-pituitary-adrenocortical system, respectively. This experimental paradigm might represent a useful model for further research on the effects of various stressors (i.e., training and competition) in taekwondo athletes. PMID:21643917

  3. Inhibitory activities against heterologous ?-amylases and in vitro allergenic reactivity of Einkorn wheats.

    PubMed

    Sánchez-Monge, R; García-Casado, G; Malpica, J M; Salcedo, G

    1996-10-01

    Salt extracts from seeds of 36 lines of Einkorn wheats were analyzed for their inhibitory activity towards two insect (Tenebrio molitor, Coleoptera, and Ephestia kuehniella, Lepidoptera) and one mammalian (human salivary) ?-amylases. Whereas all ten T. monococcum accessions tested were active towards the lepidopteran enzyme, they had no effect on the coleopteran or the mammalian ones. More variability was found among the 21 lines of T. boeticum analyzed, although none of them inhibited human ?-amylase. The five accessions of T. urartu showed even greater diversity. Among all Einkorn accessions tested, only two urartu lines affected the three ?-amylases. These lines displayed inhibition patterns similar to those of T. aestivum and T. turgidum cultivars. Since several breadwheat ?-amylase inhibitors are major allergens associated with baker's asthma, we also studied the in vitro allergenic activity of salt extracts from the Einkorn wheats under study. No significant differences in IgE-binding were found between these accessions and theT. aestivum or T. turgidum cultivars. Furthermore, putative allergens with molecular sizes in the range of 20-60 kDa were detected in these Einkorn wheats. PMID:24162403

  4. MALTOTRIOSE, PRODUCT OF ALPHA-AMYLASE STARCH HYDROLYSIS, SUPPRESSES MALTASE-GLUCOAMYLASE ACTIVITY AND SLOWS TERMINAL STARCH DIGESTION 44.5 FOLD

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Starches constitute the main caloric source in the average human diet. The digestion of starches is far more complex than sugars and requires six different enzyme activities to produce free glucose before absorption. Salivary and pancreatic alpha-amylase activities initially hydrolyze internal 1-4 g...

  5. Salivary cortisol and alpha-amylase levels during an assessment procedure correlate differently with risk-taking measures in male and female police recruits

    PubMed Central

    van den Bos, Ruud; Taris, Ruben; Scheppink, Bianca; de Haan, Lydia; Verster, Joris C.

    2013-01-01

    Recent laboratory studies have shown that men display more risk-taking behavior in decision-making tasks following stress, whilst women are more risk-aversive or become more task-focused. In addition, these studies have shown that sex differences are related to levels of the stress hormone cortisol (indicative of activation of the hypothalamus-pituitary-adrenocortical-axis): the higher the levels of cortisol the more risk-taking behavior is shown by men, whereas women generally display more risk-aversive or task-focused behavior following higher levels of cortisol. Here, we assessed whether such relationships hold outside the laboratory, correlating levels of cortisol obtained during a job-related assessment procedure with decision-making parameters in the Cambridge Gambling Task (CGT) in male and female police recruits. The CGT allows for discriminating different aspects of reward-based decision-making. In addition, we correlated levels of alpha-amylase [indicative of activation of the sympatho-adrenomedullary-axis (SAM)] and decision-making parameters. In line with earlier studies men and women only differed in risk-adjustment in the CGT. Salivary cortisol levels correlated positively and strongly with risk-taking measures in men, which was significantly different from the weak negative correlation in women. In contrast, and less strongly so, salivary alpha-amylase levels correlated positively with risk-taking in women, which was significantly different from the weak negative correlation with risk-taking in men. Collectively, these data support and extend data of earlier studies indicating that risky decision-making in men and women is differently affected by stress hormones. The data are briefly discussed in relation to the effects of stress on gambling. PMID:24474909

  6. Activity and storage of commercial amylases in the 2013 Louisiana grinding season

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A current problem in the application of amylases at sugarcane factories is the existence of a wide variation in the activities and activity per unit cost of commercial amylases. The efficiency of amylase action to break down starch in the factory is related to the activity of the amylase used. Until...

  7. Asymmetry in children's salivary cortisol and alpha-amylase in the context of marital conflict: links to children's emotional security and adjustment.

    PubMed

    Koss, Kalsea J; George, Melissa R W; Cummings, E Mark; Davies, Patrick T; El-Sheikh, Mona; Cicchetti, Dante

    2014-05-01

    Recent research supports the promise of examining interactive models of physiological processes on children's adjustment. The present study investigates interactions between children's autonomic nervous system activity and adrenocortical functioning in the context of marital discord; specifically, testing models of concurrent responses proposed by Bauer et al. ([2002] Developmental and Behavioral Pediatrics 23:102-113) in the prediction of children's behavioral responses to conflict and adjustment. Asymmetry and symmetry in children's salivary alpha-amylase and cortisol were examined in 195 children (M age?=?8 years) in response to viewing conflict vignettes. Results were partially consistent with an interactive model in the context of high marital discord; asymmetry among higher alpha-amylase and lower cortisol related to higher emotional insecurity and concurrent and subsequent maladjustment. In contrast, patterns of symmetrical responses were related to greater maladjustment for children exposed to lower levels of marital discord, supporting an additive model. Findings support the importance of a multisystem approach to investigating the adaptiveness of children's physiological stress responses, while also highlighting the value of considering physiological responses in the context of family risk. PMID:24037991

  8. Salivary alpha-amylase and cortisol in infancy and toddlerhood: direct and indirect relations with executive functioning and academic ability in childhood.

    PubMed

    Berry, Daniel; Blair, Clancy; Willoughby, Michael; Granger, Douglas A

    2012-10-01

    Using data from a predominantly low-income, population-based prospective longitudinal sample of 1292 children followed from birth, indicators of children's autonomic (salivary alpha-amylase; sAA) and hypothalamic-pituitary-adrenal (HPA) axis (salivary cortisol) activity at 7, 15, and 24 months of age were found to predict executive functioning at 36-months and academic achievement in pre-kindergarten. The findings suggested that the respective cortisol and sAA effects on executive functioning and academic achievement were interactive. Optimal developmental outcomes were associated with asymmetrical cortisol/sAA profiles. Higher cortisol levels were predictive of lower executive functioning and academic abilities, but only for those with concurrently moderate to high levels of sAA. In contrast, higher sAA concentrations were predictive of better executive functioning and academic abilities, but only for those with concurrently moderate to low levels of cortisol. These relations were statistically identical across infancy and toddlerhood. The conditional effects of cortisol and sAA on pre-kindergarten academic achievement were mediated fully by links between these early physiological indicators and executive functioning. PMID:22472478

  9. Effect of combined cognitive-behavioural therapy and endurance training on cortisol and salivary alpha-amylase in panic disorder.

    PubMed

    Plag, Jens; Gaudlitz, Katharina; Schumacher, Sarah; Dimeo, Fernando; Bobbert, Thomas; Kirschbaum, Clemens; Ströhle, Andreas

    2014-11-01

    Current data point to an alteration of both the hypothalamo-pituitary-adrenal (HPA)-system and the peripheral transmission of catecholamines in anxiety disorders. There is also some evidence for the effect of several components of cognitive-behavioural interventions such as coping and control and for an effect of exercise training on the neuroendocrine stress response in healthy subjects as well as patients suffering from distinct (mental) disorders. This double-blind, controlled study investigated the effect of cognitive-behavioural therapy (CBT) in combination with either high-level endurance training or low-level exercise on salivary cortisol (sC) and on levels of salivary alpha-amylase (sAA) in patients suffering from panic disorder (PD) with and without agoraphobia. In comparison to the low-level exercise condition, there were significantly lower sC-levels in the experimental group performing high-level endurance training at a 7-month follow-up. In contrast, there were no group differences in sAA levels during the study period. In this trial, we found evidence for a decelerated effect of endurance-training on HPA-system's functioning in PD. Further studies addressing the alteration of sAA levels in this population might investigate physical exercise different in intensity and duration. PMID:25085607

  10. The potato amylase inhibitor gene SbAI regulates cold-induced sweetening in potato tubers by modulating amylase activity.

    PubMed

    Zhang, Huiling; Liu, Jun; Hou, Juan; Yao, Ying; Lin, Yuan; Ou, Yongbin; Song, Botao; Xie, Conghua

    2014-09-01

    Potato cold-induced sweetening (CIS) is critical for the postharvest quality of potato tubers. Starch degradation is considered to be one of the key pathways in the CIS process. However, the functions of the genes that encode enzymes related to starch degradation in CIS and the activity regulation of these enzymes have received less attention. A potato amylase inhibitor gene known as SbAI was cloned from the wild potato species Solanum berthaultii. This genetic transformation confirmed that in contrast to the SbAI suppression in CIS-resistant potatoes, overexpressing SbAI in CIS-sensitive potatoes resulted in less amylase activity and a lower rate of starch degradation accompanied by a lower reducing sugar (RS) content in cold-stored tubers. This finding suggested that the SbAI gene may play crucial roles in potato CIS by modulating the amylase activity. Further investigations indicated that pairwise protein-protein interactions occurred between SbAI and ?-amylase StAmy23, ?-amylases StBAM1 and StBAM9. SbAI could inhibit the activities of both ?-amylase and ?-amylase in potato tubers primarily by repressing StAmy23 and StBAM1, respectively. These findings provide the first evidence that SbAI is a key regulator of the amylases that confer starch degradation and RS accumulation in cold-stored potato tubers. PMID:24985879

  11. Salivary pellicles on titanium and their effect on metabolic activity in Streptococcus oralis

    PubMed Central

    2013-01-01

    Background Titanium implants in the oral cavity are covered with a saliva-derived pellicle to which early colonizing microorganisms such as Streptococcus oralis can bind. The protein profiles of salivary pellicles on titanium have not been well characterized and the proteins of importance for binding are thus unknown. Biofilm bacteria exhibit different phenotypes from their planktonic counterparts and contact with salivary proteins may be one factor contributing to the induction of changes in physiology. We have characterized salivary pellicles from titanium surfaces and investigated how contact with uncoated and saliva-coated titanium surfaces affects metabolic activity in adherent cells of S. oralis. Methods Salivary pellicles on smooth titanium surfaces were desorbed and these, as well as purified human saliva, were subjected to two-dimensional gel electrophoresis and mass spectroscopy. A parallel plate flow-cell model was used to study binding of a fresh isolate of S. oralis to uncoated and saliva-coated titanium surfaces. Metabolic activity was assessed using the BacLight CTC Vitality Kit and confocal scanning laser microscopy. Experiments were carried out in triplicate and the results analyzed using Student’s t-test or ANOVA. Results Secretory IgA, ?-amylase and cystatins were identified as dominant proteins in the salivary pellicles. Selective adsorption of proteins was demonstrated by the enrichment of prolactin-inducible protein and absence of zinc-?2-glycoprotein relative to saliva. Adherence of S. oralis to titanium led to an up-regulation of metabolic activity in the population after 2 hours. In the presence of a salivary pellicle, this effect was enhanced and sustained over the following 22 hour period. Conclusions We have shown that adherence to smooth titanium surfaces under flow causes an up-regulation of metabolic activity in the early oral colonizer S. oralis, most likely as part of an adaptation to the biofilm mode of life. The effect was enhanced by a salivary pellicle containing sIgA, ?-amylase, cystatins and prolactin-inducible protein which was, for the first time, identified as an abundant component of salivary pellicles on titanium. Further studies are needed to clarify the mechanisms underlying the effect of surface contact on metabolic activity as well as to identify the salivary proteins responsible for enhancing the effect. PMID:23866104

  12. Pouteria ramiflora extract inhibits salivary amylolytic activity and decreases glycemic level in mice.

    PubMed

    De Gouveia, Neire M; De Albuquerque, Cibele L; Espindola, Laila S; Espindola, Foued S

    2013-09-01

    In this study, extracts of plant species from the Cerrado biome were assessed in order to find potential inhibitors of human salivary alpha-amylase. The plants were collected and extracts were obtained from leaves, bark, and roots. We performed a preliminary phytochemical analysis and a screening for salivar alpha-amylase inhibitory activity. Only three botanical families (Sapotaceae, Sapindaceae and Flacourtiaceae) and 16 extracts showed a substantial inhibition (>75%) of alpha-amylase. The ethanolic extracts of Pouteria ramiflora obtained from stem barks and root barks decreased amylolytic activity above 95% at a final concentration of 20 µg/mL. Thus, adult male Swiss mice were treated orally with P. ramiflora in acute toxicity and glycemic control studies. Daily administration with 25, 50 and 100 mg/kg of aqueous extract of P. ramiflora for eight days can reduce significantly body weight and blood glucose level in mice. These data suggest that the crude polar extract of P. ramiflora decreases salivary amylolytic activity while lowering the blood levels of glucose. PMID:24068095

  13. Effect of chemicals on fungal alpha-amylase activity.

    PubMed

    Ali, F S; Abdel-Moneim, A A

    1989-01-01

    The effect of 8 growth regulators at concentrations of 1,000, 5,000 and 10,000 ppm on the activity of fungal (Aspergillus flavus var. columnaris) alpha-amylase was studied. Indol acetic acid (IAA) and naphthalene acetic acid (NAA) inhibited alpha-amylase activity by 2% and 7% at 1,000 ppm. The other 6 growth regulators, indol butyric acid (IBA), gibberellic acid, cumarin, cycocel (CCC), atonik-G and kylar, did not inhibit but stimulated alpha-amylase activity (0 to 9%) at 1,000 ppm. All growth regulators studied inhibited alpha-amylase activity at 5,000 and 10,000 ppm concentration except kylar. The effect of organic acids and formaldehyde at 0.01, 0.005, and 0.001 M was studied. Acetic acid stimulated alpha-amylase at all concentrations, but formic acid, oxalic acid, lactic acid and citric acid inhibited alpha-amylase activity by 91, 100, 100 and 79%, respectively, at a concentration of 0.01 M, while by 31, 100, 15 and 20%, respectively, at 0.005 M. Formaldehyde induced 7, 3 and 2% inhibition at 0.01, 0.005 and 0.001 M, respectively. At 0.01 M either sorbitol or fructose inhibited alpha-amylase by 8%, Maltose 7%, sucrose 6%, phenol, glucose and galactose each by 5%, ethanol, glycerol, arabinose and sodium benzoate each by 4%, isopropanol and mannitol 1%, but methanol and ammonium citrate dibasic did not inhibit alpha-amylase. The results indicate that CuCl2, SnCl2, AgNO3 and Fe2(SO4)3 were the strongest inhibitors, followed by Cd(C2H3O2), HgCl2, Na2-EDTA, Na2HPO4, and CaCl2 in decreasing order. NaCl, NaBr and Mn SO4 did not inhibit alpha-amylase at concentrations from 10 mM to 0.01 mM. PMID:2515680

  14. Relationship of Salivary Alpha Amylase and Cortisol to Social Anxiety in Healthy Children Undergoing Laboratory Pain Tasks

    PubMed Central

    Payne, Laura A; Hibel, Leah C; Granger, Douglas A; Tsao, Jennie C I; Zeltzer, Lonnie K

    2014-01-01

    Objective Salivary alpha amylase (sAA) has been shown to be a sensitive and reliable marker of the autonomic nervous system (ANS) response to stress. A link between sAA, cortisol, and social/evaluative stress has been established in youth, but little is known about these relationships in response to other stressors in children, and how social anxiety might moderate these relationships. The current study explored the associations among sAA and salivary cortisol responses to laboratory pain tasks and self-reported social anxiety symptoms in a sample of healthy children. Method Two hundred thirty-one children (114 girls; 49.4%) with a mean age 12.68 years (SD=3.0; range 7–18) participated in the study. Participants completed self-report questionnaires prior to undergoing a series of laboratory pain tasks involving cold, pressure, and heat pain. Saliva samples were collected upon arrival to the laboratory (pre-task), following the completion of the pain tasks (post-task1), and 20 minutes after the completion of the pain tasks (post-task2). Results Demographic factors (age, sex, pubertal stage) did not predict either sAA or cortisol levels. However, children reporting higher levels of social anxiety demonstrated significantly higher sAA but not cortisol levels across three salivary collection times, compared to children reporting lower levels of social anxiety. Further, it does not appear that reduced state levels of anxiety before or during the tasks buffer this relationship. Conclusion These data highlight the possibility of identifying biomarkers of stress that are consistent across time and developmental stage. sAA appears to be a marker of stress response in children with self-reported social anxiety. There may also be a potentially unique relationship of sAA to stress in this population. In addition, sAA may reflect stable individual differences in levels of ANS arousal and may be a useful biomarker for identifying children at risk for stress. PMID:25525630

  15. Halotolerant Ability and ?-Amylase Activity of Some Saltwater Fungal Isolates

    PubMed Central

    Niknejad, Farhad; Moshfegh, Mahsa; Najafzadeh, Mohammad Javad; Houbraken, Jos; Rezaei, Shahla; Zarrini, Gholamreza; Faramarzi, Mohammad Ali; Nafissi-Varcheh, Nastaran

    2013-01-01

    Four halotolerant fungal isolates originating from the saltwater Lake Urmia in Iran were selected during a screening program for salt resistance and ?-amylase activity. The isolates were identified based on sequencing the ITS region and a part of the ?-tubulin gene, as Penicillium chrysogenum (isolate U1; CBS 132820), Fusarium incarnatum (isolate U2; CBS 132821), and Penicillium polonicum (isolate U3; CBS 132822, and isolate U4; CBS 132823). The growth of these isolates was determined by measuring the colony diameter and mycelia dry weight in Sabouraud dextrose agar and yeast nitrogen base medium supplemented with NaCl, KCl, and LiCl. Isolate U4 showed a growth up in 15% NaCl and U1 was the only isolate that could grow in 20% KCl. None of the strains grew in a media containing LiCl. The salt supplemented medium did not increase the size of colony diameter in all isolates (p > 0.05). The ability of the selected isolates for amylase production was quantitatively tested and showed that P. polonicum isolate U4 was the most potent producer of amylase with a yield of 260.9 U/L after 60 h, whereas P. polonicum isolate U3 was the lowest one with a production level of 97.9 U/L after 48 h. P. polonicum isolate U4 could be a suitable candidate for production of amylase on an industrial scale after optimization. PMID:24250679

  16. The activity of selected glycosidases in salivary gland tumors.

    PubMed

    Bierc, Marcin; Minarowski, Lukasz; Wo?niak, Lukasz; Chojnowska, Sylwia; Knas, Malgorzata; Szajda, Slawomir; Zwierz, Krzysztof

    2010-09-30

    The monitoring of the patients after salivary gland tumors surgery is an important clinical issue. Still imperfect diagnostic procedures also remain a challenge for searching new sensitive and specific biomarkers of neoplastic processes in salivary glands. The aim of the presented study was an the assessment of the activity of HEX, with its isoforms HEX-A and HEX-B, GLU, GAL, MAN and FUC in salivary gland tumor tissues in comparison to a healthy salivary gland tissues taken during autopsy. A group of 42 patients with benign and malignant salivary gland tumors, aged 25-65 were examined. Fragments of salivary gland tumor tissue, fragments of healthy tissue removed during autopsy, blood serum and saliva were collected from patients with salivary gland tumors and healthy volunteers. In salivary gland tumor tissue the activity of HEX, HEX-A, HEX-B, GAL, FUC was considerably higher than in comparison to healthy salivary gland tissue and ascending trend of activity of GLU, MAN was also noticed. The activity of all lysosomal exoglycosidases in blood serum in patients with salivary gland tumors was considerably higher in comparison to healthy volunteers blood serum. The considerably higher activity of HEX, HEX-A, GLU, GAL, MAN, FUC and descending trend of activity of HEX-B were noticed in saliva of patients with salivary gland tumors in comparison to healthy volunteers. The assessment of HEX in blood serum and saliva of patients with salivary gland tumor can be possibly used in diagnostics and monitoring of salivary glands tumors. PMID:21071355

  17. Individual differences in the cortisol and salivary ?-amylase awakening responses in early childhood: relations to age, sex, and sleep.

    PubMed

    Bright, Melissa A; Frick, Janet E; Out, Dorothee; Granger, Douglas A

    2014-09-01

    Recent studies have examined post-waking changes in cortisol as a marker of HPA functioning, but questions remain about the stability of this response, as well as its relation to sleep and other ANS markers. The purposes of this study were to a) examine the presence and developmental changes in the cortisol awakening response (CAR) and salivary ?-amylase awakening (sAA-AR) in a toddler sample and b) determine whether and how sleep relates to these responses in this age group. We measured cortisol and sAA upon awakening (and 30?min post-waking) and sleep characteristics using actigraphy (e.g., total sleep time, sleep efficiency, number of awakenings) in toddlers (N?=?47; 36% female, ages 12-24 months). Forty-six percent of toddlers demonstrated a CAR and 52% demonstrated a sAA-AR. Strength of either response did not change linearly with age. Additionally, likelihood of demonstrating the CAR and sAA-AR was unrelated to age, sex, awakening time, time between samples, and time since feeding. Higher waking cortisol levels were associated with a shorter total sleep time and an earlier awakening. No associations were observed between sleep characteristics and the sAA-AR, ps?>?.05. Our findings suggest that these awakening responses function independently of sleep in toddlers. Additionally, the lack of change in percentage of children showing a CAR or sAA-AR across these ages suggests that these responses are stable and not emerging reliably across the second year of life. PMID:24604597

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

    ERIC Educational Resources Information Center

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

    2012-01-01

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

  19. Effects of Hatha Yoga on Blood Pressure, Salivary ?-Amylase, and Cortisol Function Among Normotensive and Prehypertensive Youth

    PubMed Central

    Mueller, Martina; Gregoski, Mathew J.; Brunner-Jackson, Brenda; McQuade, Lisa; Matthews, Cameron; Treiber, Frank A.

    2014-01-01

    Abstract Objective: Evidence is accumulating, predominantly among clinical trials in adults, that yoga improves blood pressure (BP) control, with downregulation of the hypothalamic–pituitary–adrenal (HPA) axis and the sympathetic nervous system (SNS) projected as underlying mechanisms. This pilot study assessed whether Hatha yoga has the potential to reduce BP among youth and whether dampening of the SNS and/or HPA activity is a likely pathway of change. Design: Thirty-one seventh graders were randomly assigned to a Hatha yoga program (HYP) or attention control (AC) music or art class. Baseline and 3-month evaluations included resting BP; overnight urine samples; and saliva collected at bedtime, upon awakening, and at 30 and 60 minutes after awakening for ?-amylase and cortisol assays. Results: Twenty-eight (14 in the HYP group and 14 in the AC group) students were assessed both before and after the intervention. BP changes from pre- to post-intervention were ?3.0/?2.0?mmHg for the HYP group and ?0.07/?0.79?mmHg for the AC group (p=0.30 and 0.57, respectively). Changes in systolic BP (SBP)/diastolic BP (DBP) for the prehypertensive (75th–94th percentiles for SBP) subgroup analyses were ?10.75/?8.25?mmHg for the HYP group (n=4) versus 1.8/1.0?mmHg for the AC group (n=5) (p for SBP=0.02; p for DBP=0.09). Although no statistically significant group differences were observed with changes in SNS or HPA awakening curves (area under curve for ?-amylase and cortisol, respectively), a small to moderate effect size was seen favoring a reduction of ?-amylase activation for the HYP group (Cohen d=0.34; prehypertensive d=0.20). Conclusions: A school-based Hatha yoga program demonstrated potential to decrease resting BP, particularly among prehypertensive youth. Reduced SNS drive may be an underlying neurohormonal pathway beneficially affected by the program. A large-scale efficacy/effectiveness randomized clinical trial is warranted. PMID:24620850

  20. Relationship between Salivary Alkaline Phosphatase Enzyme Activity and The Concentrations of Salivary Calcium and Phosphate Ions.

    PubMed

    Jazaeri, Mina; Malekzadeh, Hosein; Abdolsamadi, Hamidreza; Rezaei-Soufi, Loghman; Samami, Mohammad

    2015-01-01

    Although salivary alkaline phosphatase (ALP) can balance deand remineralization processes of enamel, there is no evidence regarding its effects on the concentrations of calcium and phosphate in saliva. The present study aims to determine the relationship between salivary ALP activity and the concentrations of calcium and phosphate in saliva. In this cross-sectional study, we evaluated salivary markers in 120 males, ages 19 to 44 years. All participants provided 5 mL of unstimulated whole saliva and the level of enzyme activity as well as calcium and phosphate concentrations were measured using a colorimetric method. Data were gathered and analyzed by statistical package for social sciences (SPSS) 13.00 using Pearson correlation test. A p value of <0.05 was considered statistically significant. The mean age of participants in the present study was 32.95 ± 8.09 years. The mean pH of saliva was 6.65 ± 0.62. Salivary parameters included average ALP activity (5.04 ± 1.866 U/dL), calcium (4.77 ± 0.877 mg/dL) and phosphate (10.38 ± 2.301 mg/dL). Pearson correlation test showed no significant relationship between ALP activity and calcium and phosphate concentrations in saliva (p>0.05). According to the results of the present study, there was no significant relation between salivary ALP activity and calcium and phosphate concentrations in saliva. However, further research is highly recommended. PMID:25870846

  1. Relationship between Salivary Alkaline Phosphatase Enzyme Activity and The Concentrations of Salivary Calcium and Phosphate Ions

    PubMed Central

    Jazaeri, Mina; Malekzadeh, Hosein; Abdolsamadi, Hamidreza; Rezaei-Soufi, Loghman; Samami, Mohammad

    2015-01-01

    Although salivary alkaline phosphatase (ALP) can balance deand remineralization processes of enamel, there is no evidence regarding its effects on the concentrations of calcium and phosphate in saliva. The present study aims to determine the relationship between salivary ALP activity and the concentrations of calcium and phosphate in saliva. In this cross-sectional study, we evaluated salivary markers in 120 males, ages 19 to 44 years. All participants provided 5 mL of unstimulated whole saliva and the level of enzyme activity as well as calcium and phosphate concentrations were measured using a colorimetric method. Data were gathered and analyzed by statistical package for social sciences (SPSS) 13.00 using Pearson correlation test. A p value of <0.05 was considered statistically significant. The mean age of participants in the present study was 32.95 ± 8.09 years. The mean pH of saliva was 6.65 ± 0.62. Salivary parameters included average ALP activity (5.04 ± 1.866 U/dL), calcium (4.77 ± 0.877 mg/dL) and phosphate (10.38 ± 2.301 mg/dL). Pearson correlation test showed no significant relationship between ALP activity and calcium and phosphate concentrations in saliva (p>0.05). According to the results of the present study, there was no significant relation between salivary ALP activity and calcium and phosphate concentrations in saliva. However, further research is highly recommended.

  2. Changes in alpha-and beta-amylase activities during seed germination of African finger millet.

    PubMed

    Gimbi, Dorothy Machunda; Kitabatake, Naofumi

    2002-11-01

    Changes in alpha- and beta-amylase activities in African finger millet (Eleusine coracana (L) Gaertener) were followed during germination. Germination on a small scale was performed at 15 degrees C for 1-10 days and at 20, 25 and 30 degrees C for 1-8 days. alpha- and beta-Amylase activities in malt crude extracts of germinated finger millet were evaluated spectrophotometrically using chromogenic methods. The highest alpha-amylase activity was exhibited in malt flour of finger millet germinated at 15 degrees C for 9 days and at 20 degrees C for 6 days, while the highest beta-amylase activity was displayed in the malt flour germinated for 5 days at 30 degrees C. Thermo-stability of these enzymes in malt extracts was also evaluated. Malt extracts incubated at 40 and 50 degrees C for up to 4 h retained about 84 and 64% of alpha-amylase activities, respectively. There was a substantial decrease in alpha-amylase activity to more than 90% when malt extracts were incubated at 70 and 90 degrees C for 40 and 10 min, respectively. beta-Amylase was completely inactivated when the crude extract was incubated at 70 degrees C for only 10 min. At pH 5.4, alpha-amylase displayed maximum catalytic activity at around 45 degrees C. Optimum temperature for beta-amylase activity at pH 6.0 was between 50 and 55 degrees C. Activity staining for alpha-amylase was also performed and three bands of activity were found in malt extract, each possibly representing an isozyme of alpha-amylase from finger millet. PMID:12590743

  3. Characterization of a Hydrophobic Amylase Inhibitor from Corn (Zea mays) Seeds with Activity Against Amylase from Fusarium verticillioides.

    PubMed

    Figueira, Edson L Z; Hirooka, Elisa Y; Mendiola-Olaya, Elizabeth; Blanco-Labra, Alejandro

    2003-08-01

    ABSTRACT A hydrophobic 19.7-kDa amylase inhibitor (AI) was purified from corn kernels by 95% ethanol extraction and anionic exchange chromatography. The AI has an isoelectric point of 3.6 and was very stable at different pH values and high temperatures, maintaining 47.6% activity after heating to 94 degrees C for 60 min. Amino acid analysis indicated high valine, leucine, glycine, alanine, and glutamic acid/glutamine content, and especially high valine content (41.2 mol%). This inhibitor is not a glycoprotein. It required 30-min preincubation to maximize complex enzyme-inhibitor formation when the amylase from Fusarium verticillioides was tested. The optimal pH of interaction was 6.5. It showed broad-spectrum activity including the following amylases: human saliva, porcine pancreas, F. verticillioides, as well as those from some insects of agricultural importance (Acanthoscelides obtectus, Zabrotes subfasciatus, Sitophilus zeamais, and Prostephanus truncatus). This novel hydrophobic protein not only inhibited the amylase from F. verticillioides but also decreased the conidia germination. Thus, this protein represents an approach to decrease the production of fumonisin in corn, either by using it as a molecular marker to detect fungal resistance or through genetic engineering. PMID:18943857

  4. Potent ?-amylase inhibitory activity of Indian Ayurvedic medicinal plants

    PubMed Central

    2011-01-01

    Background Indian medicinal plants used in the Ayurvedic traditional system to treat diabetes are a valuable source of novel anti-diabetic agents. Pancreatic ?-amylase inhibitors offer an effective strategy to lower the levels of post-prandial hyperglycemia via control of starch breakdown. In this study, seventeen Indian medicinal plants with known hypoglycemic properties were subjected to sequential solvent extraction and tested for ?-amylase inhibition, in order to assess and evaluate their inhibitory potential on PPA (porcine pancreatic ?-amylase). Preliminary phytochemical analysis of the lead extracts was performed in order to determine the probable constituents. Methods Analysis of the 126 extracts, obtained from 17 plants (Aloe vera (L.) Burm.f., Adansonia digitata L., Allium sativum L., Casia fistula L., Catharanthus roseus (L.) G. Don., Cinnamomum verum Persl., Coccinia grandis (L.) Voigt., Linum usitatisumum L., Mangifera indica L., Morus alba L., Nerium oleander L., Ocimum tenuiflorum L., Piper nigrum L., Terminalia chebula Retz., Tinospora cordifolia (Willd.) Miers., Trigonella foenum-graceum L., Zingiber officinale Rosc.) for PPA inhibition was initially performed qualitatively by starch-iodine colour assay. The lead extracts were further quantified with respect to PPA inhibition using the chromogenic DNSA (3, 5-dinitrosalicylic acid) method. Phytochemical constituents of the extracts exhibiting? 50% inhibition were analysed qualitatively as well as by GC-MS (Gas chromatography-Mass spectrometry). Results Of the 126 extracts obtained from 17 plants, 17 extracts exhibited PPA inhibitory potential to varying degrees (10%-60.5%) while 4 extracts showed low inhibition (< 10%). However, strong porcine pancreatic amylase inhibitory activity (> 50%) was obtained with 3 isopropanol extracts. All these 3 extracts exhibited concentration dependent inhibition with IC50 values, viz., seeds of Linum usitatisumum (540 ?gml-1), leaves of Morus alba (1440 ?gml-1) and Ocimum tenuiflorum (8.9 ?gml-1). Acarbose as the standard inhibitor exhibited an IC50 (half maximal inhibitory concentration)value of 10.2 ?gml-1. Phytochemical analysis revealed the presence of alkaloids, tannins, cardiac glycosides, flavonoids, saponins and steroids with the major phytoconstituents being identified by GC-MS. Conclusions This study endorses the use of these plants for further studies to determine their potential for type 2 diabetes management. Results suggests that extracts of Linum usitatisumum, Morus alba and Ocimum tenuiflorum act effectively as PPA inhibitors leading to a reduction in starch hydrolysis and hence eventually to lowered glucose levels. PMID:21251279

  5. Green tea consumption after intense taekwondo training enhances salivary defense factors and antibacterial capacity.

    PubMed

    Lin, Shiuan-Pey; Li, Chia-Yang; Suzuki, Katsuhiko; Chang, Chen-Kang; Chou, Kuei-Ming; Fang, Shih-Hua

    2014-01-01

    The aim of this study was to investigate the short-term effects of green tea consumption on selected salivary defense proteins, antibacterial capacity and anti-oxidation activity in taekwondo (TKD) athletes, following intensive training. Twenty-two TKD athletes performed a 2-hr TKD training session. After training, participants ingested green tea (T, caffeine 6 mg/kg and catechins 22 mg/kg) or an equal volume of water (W). Saliva samples were collected at three time points: before training (BT-T; BT-W), immediately after training (AT-T; AT-W), and 30 min after drinking green tea or water (Rec-T; Rec-W). Salivary total protein, immunoglobulin A (SIgA), lactoferrin, ?-amylase activity, free radical scavenger activity (FRSA) and antibacterial capacity were measured. Salivary total protein, lactoferrin, SIgA concentrations and ?-amylase activity increased significantly immediately after intensive TKD training. After tea drinking and 30 min rest, ?-amylase activity and the ratio of ?-amylase to total protein were significantly higher than before and after training. In addition, salivary antibacterial capacity was not affected by intense training, but green tea consumption after training enhanced salivary antibacterial capacity. Additionally, we observed that salivary FRSA was markedly suppressed immediately after training and quickly returned to pre-exercise values, regardless of which fluid was consumed. Our results show that green tea consumption significantly enhances the activity of ?-amylase and salivary antibacterial capacity. PMID:24498143

  6. Green Tea Consumption after Intense Taekwondo Training Enhances Salivary Defense Factors and Antibacterial Capacity

    PubMed Central

    Lin, Shiuan-Pey; Li, Chia-Yang; Suzuki, Katsuhiko; Chang, Chen-Kang; Chou, Kuei-Ming; Fang, Shih-Hua

    2014-01-01

    The aim of this study was to investigate the short-term effects of green tea consumption on selected salivary defense proteins, antibacterial capacity and anti-oxidation activity in taekwondo (TKD) athletes, following intensive training. Twenty-two TKD athletes performed a 2-hr TKD training session. After training, participants ingested green tea (T, caffeine 6 mg/kg and catechins 22 mg/kg) or an equal volume of water (W). Saliva samples were collected at three time points: before training (BT-T; BT-W), immediately after training (AT-T; AT-W), and 30 min after drinking green tea or water (Rec-T; Rec-W). Salivary total protein, immunoglobulin A (SIgA), lactoferrin, ?-amylase activity, free radical scavenger activity (FRSA) and antibacterial capacity were measured. Salivary total protein, lactoferrin, SIgA concentrations and ?-amylase activity increased significantly immediately after intensive TKD training. After tea drinking and 30 min rest, ?-amylase activity and the ratio of ?-amylase to total protein were significantly higher than before and after training. In addition, salivary antibacterial capacity was not affected by intense training, but green tea consumption after training enhanced salivary antibacterial capacity. Additionally, we observed that salivary FRSA was markedly suppressed immediately after training and quickly returned to pre-exercise values, regardless of which fluid was consumed. Our results show that green tea consumption significantly enhances the activity of ?-amylase and salivary antibacterial capacity. PMID:24498143

  7. a-Amylase activity during pullulan production and a-Amylase gene analyses of Aureobasidium pullulans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fungus Aureobasidium pullulans is the source of commercially produced pullulan, a high molecular weight polysaccharide that is used in the manufacture of edible films. It has been proposed that alpha-amylase negatively affects the molecular weight of pullulan in late cultures. Based on a recen...

  8. Studies on the Utility of ß-amylase1 IntronIII Sequences as Markers for ß-amylase Activity and Thermostability, Diastatic Power and Malt Quality

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The third intron of barley (Hordeum vulgare L.) ß-amylase 1 (Bmy1) is extremely polymorphic. The use of specific insertion/deletions (indels) in the third intron as markers for cultivar development has been recommended based on associations with ß-amylase activity and thermostability. The third intr...

  9. Inhibition of ?-amylase activity by cellulose: Kinetic analysis and nutritional implications.

    PubMed

    Dhital, Sushil; Gidley, Michael J; Warren, Frederick J

    2015-06-01

    We report on inhibition of ?-amylase activity by cellulose based on in vitro experiments. The presence of cellulose in the hydrolysing medium reduced the initial velocity of starch hydrolysis in a concentration dependent manner. ?-Amylase adsorption to cellulose was reversible, attaining equilibrium within 30min of incubation, and showed a higher affinity at 37°C compared to 20 and 0°C. The adsorption was almost unchanged in the presence of maltose (2.5-20mM) but was hindered in the presence of excess protein, suggesting non-specific adsorption of ?-amylase to cellulose. Kinetic analyses of ?-amylase hydrolysis of maize starch in the presence of cellulose showed that the inhibition is of a mixed type. The dissociation constant (Kic) of the EI complex was found to be ca. 3mg/mL. The observed inhibition of ?-amylase activity suggests that cellulose in the diet can potentially attenuate starch hydrolysis. PMID:25843863

  10. In vitro study on ?-amylase inhibitory activity of an Indian medicinal plant, Phyllanthus amarus

    PubMed Central

    Tamil, Iniyan G.; Dineshkumar, B.; Nandhakumar, M.; Senthilkumar, M.; Mitra, A.

    2010-01-01

    Objective: The objective of this study was to evaluate the ?-amylase inhibitory activity of different extracts of Phyllanthus amarus against porcine pancreatic amylase in vitro. Materials and Methods: The plant extracts were prepared sequentially with ethanol, chloroform, and hexane. Each extract was evaporated using rotary evaporator, under reduced pressure. Different concentrations (10, 20, 40, 60, 80, and 100 ?g/mL) of each extract were made by using dimethyl sulfoxide (DMSO) and subjected to ?-amylase inhibitory assay using starch azure as a substrate. The absorbance was read at 595 nm using spectrophotometer. Using this method, the percentage of ?-amylase inhibitory activity and IC50values of each extract was calculated. Results: The chloroform extract failed to inhibit ?-amylase activity. However, the ethanol and hexane extracts of P. amarus exhibited appreciable ?-amylase inhibitory activity with an IC50 values 36.05 ± 4.01 ?g/mL and 48.92 ± 3.43 ?g/mL, respectively, when compared with acarbose (IC50value 83.33 ± 0.34 ?g/mL). Conclusion: This study supports the ayurvedic concept that ethanol and hexane extracts of P. amarus exhibit considerable ?-amylase inhibitory activities. Further, this study supports its usage in ethnomedicines for management of diabetes. PMID:21206618

  11. Regulation and cloning of the gene encoding amylase activity of the ruminal bacterium Streptococcus bovis.

    PubMed Central

    Cotta, M A; Whitehead, T R

    1993-01-01

    Streptococcus bovis is an important starch-degrading ruminal bacterium that has been implicated as being important in the etiology of a number of ruminal pathologies associated with diets high in grains. Previous studies with S. bovis have shown that amylase production was influenced by the growth substrate, but the nature of this regulation was not determined. The current study was conducted to better describe the regulatory phenomena and gain a better understanding of the molecular characteristics of this activity. Nutritional experiments demonstrated that the presence of starch or the starch-derived disaccharide maltose was required for maximum amylase production. Subsequent time-course experiments showed that amylase synthesis was induced by maltose and repressed by glucose, cellobiose, and fructose, while inulin and lactose had little effect on enzyme accumulation. The effects of the added antibiotics rifampin and tetracycline were consistent with transcriptional control of amylase synthesis. Analysis of S. bovis cells grown on glucose or maltose showed that they contained similar low levels of cyclic AMP, indicating that it was unlikely that regulation of amylase synthesis was mediated through a mechanism involving this nucleotide. The amylase gene from S. bovis JB1 was cloned and expressed in Escherichia coli. The amylase produced in E. coli was of lower molecular weight than that synthesized by S. bovis and had catalytic characteristics different from those of S. bovis amylase. When the gene was introduced back into S. bovis JB1, only one form of amylase activity was detected, indicating that the entire gene was present on this insert. The use of the amylase gene as a genetic probe for identification of S. bovis strains is discussed. Images PMID:7679887

  12. Relation of amylase to starch and Lycasin metabolism in human dental plaque in vitro.

    PubMed

    Birkhed, D; Skude, G

    1978-07-01

    Acid production activity (APA) in plaque suspensions from glucose, boiled soluble starch and hydrogenated starch hydrolysate (Lycasin) was studied in 11 subjects. Amylase (alpha-1,4-glucan 4-glucanohydrolase, EC 3.2.1.1) activity was measured in plaque and whole saliva samples from the same persons. Lycasin was found to be hydrolyzed by salivary amylase under the formation of di- and oligosaccharides, however, with a lower rate than starch. A high correlation was found between APA from glucose and from soluble starch and between APA from soluble starch and plaque amylase activity. No correlation was found between amylase activity in saliva and APA from soluble starch or between amylase activity in saliva or plaque and APA from Lycasin. APA from Lycasin was about 62% and from soluble starch about 76% of the APA from glucose. 0-25% of the total number of cultivable microorganisms from the plaque produced extracellular starch-degrading enzymes. No correlation was found between number of starch-degrading microorganisms and APA from soluble starch or between these numbers and the plaque amylase activity. By electrophoreses only amylase fractions of human origin were found in whole saliva, plaque supernatants and plaque suspensions, indicating that the microbial amylase activity in the plaque is low compared with that of salivary origin. PMID:279956

  13. Inhibition of ?-Amylase and ?-Glucosidase Activity by Tea and Grape Seed Extracts and their Constituent Catechins

    PubMed Central

    Yilmazer-Musa, Meltem; Griffith, Anneke M.; Michels, Alexander J.; Schneider, Erik; Frei, Balz

    2015-01-01

    We evaluated the inhibitory effects of plant-based extracts (grape seed, green tea, and white tea) on ?-amylase and ?-glucosidase activity, glucosidases required for starch digestion. The abundant flavan-3-ol monomers (catechins) in these extracts were also tested for their inhibitory potential and evaluated against the pharmacological glucosidase inhibitor, acarbose. To evaluate relative potency of these extracts and catechins, the concentrations required for 50 and 90% inhibition of enzyme activity were determined. Maximum enzyme inhibition was used to assess an inhibitor’s relative efficacy. Results showed that grape seed extract strongly inhibited both ?-amylase and ?-glucosidase activity, with equal and much higher potency, respectively, than acarbose. While tea extracts and individual catechin 3-gallates were less effective inhibitors of ?-amylase, they were potent inhibitors of ?-glucosidase. Our data show that plant extracts containing catechin 3-gallates are potent inhibitors of ?-glucosidase, and suggest that procyanidins found in grape seed extract strongly inhibit ?-amylase activity. PMID:22697360

  14. Inhibitory activity of ?-amylase and ?-glucosidase by plant extracts from the Brazilian cerrado.

    PubMed

    Souza, Paula Monteiro de; Sales, Paloma Michelle de; Simeoni, Luiz Alberto; Silva, Elton Clementino; Silveira, Dâmaris; Magalhães, Pérola de Oliveira

    2012-03-01

    Diabetes mellitus is the most common disease in the world. One therapeutic approach for treating diabetes is inhibition of ?-amylase and ?-glucosidase activities to reduce postprandial blood glucose levels. In vitro tests showed that several plant extracts from Brazilian cerrado species can inhibit the activity of ?-amylase and ?-glucosidase. The extracts of Eugenia dysenterica, Stryphnodendron adstringens, Pouteria caimito, Pouteria ramiflora, and Pouteria torta showed strong ?-amylase and ?-glucosidase inhibitory activity. Eugenia dysenterica, P. caimito, P. ramiflora, and P. torta aqueous extracts exerted the highest activity against ?-amylase (IC??) values of 14.93, 13.6, 7.08, and 5.67 µg/mL, respectively) and ?-glucosidase (IC?? values of 0.46, 2.58, 0.35, and 0.22 µg/mL, respectively). Stryphnodendron adstringens ethanol extract also exhibited inhibitory activity against both enzymes (IC??) 1.86 µg/mL against ?-amylase and 0.61 µg/mL against ?-glucosidase). The results suggest that the activity of these cerrado plants on ?-amylase and ?-glucosidase represents a potential tool for development of new strategies for treatment of diabetes. PMID:22134849

  15. Amylase-Binding Protein B of Streptococcus gordonii Is an Extracellular Dipeptidyl-Peptidase?

    PubMed Central

    Chaudhuri, Biswendu; Paju, Susanna; Haase, Elaine M.; Vickerman, M. Margaret; Tanzer, Jason M.; Scannapieco, Frank A.

    2008-01-01

    The oral commensal bacterium Streptococcus gordonii interacts with salivary amylase via two amylase-binding proteins, AbpA and AbpB. Based on sequence analysis, the 20-kDa AbpA protein is unique to S. gordonii, whereas the 82-kDa AbpB protein appears to share sequence homology with other bacterial dipeptidases. The aim of this study was to verify the peptidase activity of AbpB and further explore its potential functions. The abpB gene was cloned, and histidine-tagged AbpB (His-AbpB) was expressed in Escherichia coli and purified. Its amylase-binding activity was verified in an amylase ligand binding assay, and its cross-reactivity was verified with an anti-AbpB antibody. Both recombinant His-AbpB and partially purified native AbpB displayed dipeptidase activity and degraded human type VI collagen and fibrinogen, but not salivary amylase. Salivary amylase precipitates not only AbpA and AbpB but also glucosyltransferase G (Gtf-G) from S. gordonii supernatants. Since Streptococcus mutans also releases Gtf enzymes that could also be involved in multispecies plaque interactions, the effect of S. gordonii AbpB on S. mutans Gtf-B activity was also tested. Salivary amylase and/or His-AbpB caused a 1.4- to 2-fold increase of S. mutans Gtf-B sucrase activity and a 3- to 6-fold increase in transferase activity. An enzyme-linked immunosorbent assay verified the interaction of His-AbpB and amylase with Gtf-B. In summary, AbpB demonstrates proteolytic activity and interacts with and modulates Gtf activity. These activities may help explain the crucial role AbpB appears to play in S. gordonii oral colonization. PMID:18678669

  16. LEADER 3—Lipase and Amylase Activity in Subjects With Type 2 Diabetes

    PubMed Central

    Steinberg, William M.; Nauck, Michael A.; Zinman, Bernard; Daniels, Gilbert H.; Bergenstal, Richard M.; Mann, Johannes F.E.; Steen Ravn, Lasse; Moses, Alan C.; Stockner, Mette; Baeres, Florian M.M.; Marso, Steven P.; Buse, John B.

    2014-01-01

    Objectives This report from the LEADER (Liraglutide Effect and Action in Diabetes: Evaluation of Cardiovascular Outcome Results) trial describes baseline lipase and amylase activity in type 2 diabetic subjects without acute pancreatitis symptoms before randomization to the glucagonlike peptide analog liraglutide or placebo. Methods The LEADER is an international randomized placebo-controlled trial evaluating the cardiovascular safety of liraglutide in 9340 type 2 diabetic patients at high cardiovascular risk. Fasting lipase and amylase activity was assessed at baseline, before receiving liraglutide or placebo, using a commercial assay (Roche) with upper limit of normal values of 63 U/L for lipase and 100 U/L for amylase. Results Either or both enzymes were above the upper limit of normal in 22.7% of subjects; 16.6% (n = 1540) had an elevated lipase level (including 1.2% >3-fold elevated), and 11.8% (n = 1094) had an elevated amylase level (including 0.2% >3-fold elevated). In multivariable regression models, severely reduced kidney function was associated with the largest effect on increasing activity of both. However, even among subjects with normal kidney function, 12.2% and 7.7% had elevated lipase and amylase levels. Conclusions In this large study of type 2 diabetic patients, nearly 25% had elevated lipase or amylase levels without symptoms of acute pancreatitis. The clinician must take these data into account when evaluating abdominal symptoms in type 2 diabetic patients. PMID:25275271

  17. Smoking increases salivary arginase activity in patients with dental implants

    Microsoft Academic Search

    D. A. Queiroz; J. R. Cortelli; M. Holzhausen; E. Rodrigues; D. R. Aquino; W. A. Saad

    2009-01-01

    It is believed that an increased arginase activity may lead to less nitric oxide production, which consequently increases\\u000a the susceptibility to bacterial infection. Considering the hypothesis that smoking may alter the arginase activity and that\\u000a smoking is considered a risk factor to dental implant survival, the present study aimed at evaluating the effect of smoking\\u000a on the salivary arginase activity

  18. Effect of ultrasound on the activity and conformation of ?-amylase, papain and pepsin.

    PubMed

    Yu, Zhi-Long; Zeng, Wei-Cai; Zhang, Wen-Hua; Liao, Xue-Pin; Shi, Bi

    2014-05-01

    The effect of ultrasound on the activity of ?-amylase, papain and pepsin was investigated and the mechanism of the effect was explored by determining their conformational changes. With the irradiation of power ultrasound, the activity of ?-amylase and papain was inhibited, while the activity of pepsin was activated. According to the analysis of circular dichroism, Fourier transform infrared and fluorescence spectroscopy, the ?o ? ?(?) amide transitions and secondary structural components, especially ?-sheet, of these three enzymes were significantly influenced by ultrasound. The tryptophan fluorescence intensity of the three enzymes was also observed to be affected by sonication. Furthermore, it was found that the pepsin molecule might gradually be resistant to prolonged ultrasonic treatment and recover from the ultrasound-induced damage to its original structure. The results suggested that the activity of ?-amylase, papain and pepsin could be modified by ultrasonic treatment mainly due to the variation of their secondary and tertiary structures. PMID:24291306

  19. Inhibition of ?-amylase and ?-glucosidase activities by ethanolic extract of Telfairia occidentalis (fluted pumpkin) leaf

    PubMed Central

    Oboh, G; Akinyemi, AJ; Ademiluyi, AO

    2012-01-01

    Objective To investigate the inhibitory effect of Telfairia occidentalis Hook f. (Curcubitaceae) (T. occidentalis) leaf on key enzyme linked to type-2 diabetes (? - amylase and ? - glucosidase) as well as assess the effect of blanching (a commonly practiced food processing technique) of the vegetable on these key enzymes. Methods Fresh leaves of T. occidentalis were blanched in hot water for 10 minutes, and the extracts of both the fresh and blanched vegetables were prepared and used for subsequent analysis. The inhibitory effect of the extract on ? - amylase and ? - glucosidase activities as well as some antioxidant parameter was determined in vitro. Results The result revealed that unprocessed T. occidentalis leaf reduce Fe3+ to Fe2+ and also inhibited ? - amylase and ? - glucosidase activities in a dose dependent manner. However, blanching of the leafy vegetables caused a significant (P<0.05) increase in the antioxidant properties but decrease their ability to inhibit ? - amylase and ? - glucosidase activities. Conclusions This antioxidant properties and enzyme inhibition could be part of the mechanism by which they are used in the treatment/prevention of type-2 diabetes. However, the blanched vegetable reduces their ability to inhibit both ? - amylase and ? - glucosidase activity in vitro. PMID:23570004

  20. Time-dependent nanogel aggregation for naked-eye assays of ?-amylase activity.

    PubMed

    Jiang, Hui; Wang, Xuemei

    2012-06-01

    This work designs an enzyme-stimulated nanogel aggregation system for the naked-eye assays of ?-amylase activity. The visible aggregation of the starch-stabilized CdTe nanogels may be accelerated by ?-amylase through its efficient cleavage of glycosidic bonds in the starch network, which has been verified by the evidences from transmission electron microscopy and dynamic light scattering spectra. The required aggregation time, as validated by both the theoretical deduction and the experimental results, is inversely proportional to the enzymatic activity. Therefore a facile method has been proposed for the detection of enzyme activity, with an excellent linear range and a low detection limit. This nanogel-based protocol can be successfully applied in the fast and accurate assays of ?-amylase activity in saliva samples with a satisfactory correlation with the standard protocol, suggesting its promising applications in the biomedical and clinical fields, especially in point-of-care testing. PMID:22534693

  1. Detection of amylase activity from fruit and vegetables in an undergraduate classroom

    Microsoft Academic Search

    Surasak Laloknam; Supaporn Sirisopana; Somkiat Phornphisutthimas

    2009-01-01

    This research aimed to construct a hands-on activity for undergraduate students to understand how to detect and compare amylase activity from various sources by using a simple method. The amylolytic activity of extracts from 10 kinds of vegetables, Chinese white vegetable, tomato, cucumber, pumpkin, pea eggplant, carrot, cabbage, morning glory, Chinese broccoli, and yard long bean as well as 10

  2. Modified alpha-amylase activity among insecticide-resistant and -susceptible strains of the maize weevil, Sitophilus zeamais.

    PubMed

    Lopes, K V G; Silva, L B; Reis, A P; Oliveira, M G A; Guedes, R N C

    2010-09-01

    Fitness cost is usually associated with insecticide resistance and may be mitigated by increased energy accumulation and mobilization. Preliminary evidence in the maize weevil (Coleoptera: Curculionidae) suggested possible involvement of amylases in such phenomenon. Therefore, alpha-amylases were purified from an insecticide-susceptible and two insecticide-resistant strains (one with fitness cost [resistant cost strain], and the other without it [resistant no-cost strain]). The main alpha-amylase of each strain was purified by glycogen precipitation and ion-exchange chromatography (>or=70-fold purification, amylase bands with the same molecular mass (53.7kDa) were revealed for each insect strain. Higher activity was obtained at 35-40 degrees C and at pH 5.0-7.0 for all of the strains. The alpha-amylase from the resistant no-cost strain exhibited higher activity towards starch and lower inhibition by acarbose and wheat amylase inhibitors. Opposite results were observed for the alpha-amylase from the resistant cost strain. Although the alpha-amylase from the resistant cost strain exhibited higher affinity to starch (i.e., lower K(m)), its V(max)-value was the lowest among the strains, particularly the resistant no-cost strain. Such results provide support for the hypothesis that enhanced alpha-amylase activity may be playing a major role in mitigating fitness costs associated with insecticide resistance. PMID:20223242

  3. General Subject 1. Report to ICUMSA on the determination of commercial alpha-amylase activity by a spectrophotometric method

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A report is given on a new industrial method for the determination of the activity or strength of commercial alpha-amylase at a sugarcane factory or refinery, as well as a recommendation. At the present time, the activities or strengths of commercial alpha-amylases cannot be directly compared becau...

  4. Smart phone: a popular device supports amylase activity assay in fisheries research.

    PubMed

    Thongprajukaew, Karun; Choodum, Aree; Sa-E, Barunee; Hayee, Ummah

    2014-11-15

    Colourimetric determinations of amylase activity were developed based on a standard dinitrosalicylic acid (DNS) staining method, using maltose as the analyte. Intensities and absorbances of red, green and blue (RGB) were obtained with iPhone imaging and Adobe Photoshop image analysis. Correlation of green and analyte concentrations was highly significant, and the accuracy of the developed method was excellent in analytical performance. The common iPhone has sufficient imaging ability for accurate quantification of maltose concentrations. Detection limits, sensitivity and linearity were comparable to a spectrophotometric method, but provided better inter-day precision. In quantifying amylase specific activity from a commercial source (P>0.02) and fish samples (P>0.05), differences compared with spectrophotometric measurements were not significant. We have demonstrated that iPhone imaging with image analysis in Adobe Photoshop has potential for field and laboratory studies of amylase. PMID:24912700

  5. ON THE ACTIVITY OF TOTAL AMYLASE IN SOME SPECIES OF THE BROMUS GENUS, DURING GERMINATION

    Microsoft Academic Search

    ELENA CIORNEA; GABRIELA VASILE; DUMITRU COJOCARU

    The study discusses the activity of total amylase in the germinated caryopses of the three species belonging to the Bromus genus, namely: Bromus squarossus, Bromus japonicus and Bromus sterilis. Statistical analysis of the experimental results obtained shows that the enzymatic activity is strongly influenced by the germination time, negligible differences being recorded from one species to another.

  6. Activity of wheat alpha-amylase inhibitors towards bruchid alpha-amylases and structural explanation of observed specificities.

    PubMed

    Franco, O L; Rigden, D J; Melo, F R; Bloch, C; Silva, C P; Grossi de Sá, M F

    2000-04-01

    Plant alpha-amylase inhibitors show great potential as tools to engineer resistance of crop plants against pests. Their possible use is, however, complicated by observed variations in specificity of enzyme inhibition, even within closely related families of inhibitors. Five alpha-amylase inhibitors of the structural 0.19 family were isolated from wheat kernels, and assayed against three insect alpha-amylases and porcine pancreatic alpha-amylase, revealing several intriguing differences in inhibition profiles, even between proteins sharing sequence identity of up to 98%. Inhibition of the enzyme from a commercially important pest, the bean weevil Acanthoscelides obtectus, is observed for the first time. Using the crystal structure of an insect alpha-amylase in complex with a structurally related inhibitor, models were constructed and refined of insect and human alpha-amylases bound to 0.19 inhibitor. Four key questions posed by the differences in biochemical behaviour between the five inhibitors were successfully explained using these models. Residue size and charge, loop lengths, and the conformational effects of a Cys to Pro mutation, were among the factors responsible for observed differences in specificity. The improved structural understanding of the bases for the 0.19 structural family inhibitor specificity reported here may prove useful in the future for the rational design of inhibitors possessing altered inhibition characteristics. PMID:10759839

  7. Effect of sunflower oil on sheep small intestinal digesta viscosity, composition and amylase activity

    Microsoft Academic Search

    P. S Mir; M Ivan; G. J Mears; B. F Benkel; C. M Ross; S. D Husar; Z Mir

    2002-01-01

    Dietary oil triggers the release of small intestinal hormones; thus we decided to determine the associated effects on digesta viscosity, total dry matter (DM), protein content and amylase activity in the small intestine. These parameters were investigated in small intestinal digesta collected 15min after slaughter from 18 sheep, finished on a diet of (% DM) barley silage, 60; barley grain,

  8. DIRECT SCREENING OF LIBRARIES OF YEAST CLONES FOR ALPHA-AMYLASE ACTIVITY ON RAW STARCH

    Technology Transfer Automated Retrieval System (TEKTRAN)

    High-throughput screening for high-activity barley alpha-amylase mutants expressed in Saccharomyces cerevisiae is hampered by the interference of the glucose used in yeast growth media. In a previous report, it was demonstrated that glycerol could be used as an alternative carbon source, with an un...

  9. Complement activation by salivary agglutinin is secretor status dependent.

    PubMed

    Gunput, Sabrina T G; Ligtenberg, Antoon J M; Terlouw, Bas; Brouwer, Mieke; Veerman, Enno C I; Wouters, Diana

    2015-01-01

    After mucosal damage or gingival inflammation, complement proteins leak into the oral cavity and mix with salivary proteins such as salivary agglutinin (SAG/gp-340/DMBT1). This protein is encoded by the gene Deleted in Malignant Brain Tumors 1 (DMBT1), and it aggregates bacteria, viruses and fungi, and activates the lectin pathway of the complement system. In the lectin pathway, carbohydrate structures on pathogens or altered self cells are recognized. SAG is highly glycosylated, partly on the basis of the donor's blood group status. Whereas secretors express Lewis b, Lewis y, and antigens from the ABO-blood group system on SAG, non-secretors do not. Through mannose-binding lectin (MBL) binding and C4 deposition assays, we aimed to identify the chemical structures on SAG that are responsible for complement activation. The complement-activating properties of SAG were completely abolished by oxidation of its carbohydrate moiety. SAG-mediated activation of complement was also inhibited in the presence of saccharides such as fucose and Lewis b carbohydrates, and also after pretreatment with the fucose-binding lectin, Anguilla anguilla agglutinin. Complement activation was significantly (p<0.01) higher in secretors than in non-secretors. Our results suggest that fucose-rich oligosaccharide sidechains, such as Lewis b antigens, are involved in the activation of complement by SAG. PMID:25153235

  10. Salivary Acetylcholinesterase Activity Is Increased in Parkinson's Disease: A Potential Marker of Parasympathetic Dysfunction

    PubMed Central

    Fedorova, Tatyana; Knudsen, Cindy Soendersoe; Mouridsen, Kim; Nexo, Ebba; Borghammer, Per

    2015-01-01

    Introduction. Decreased salivary flow and xerostomia are frequent findings in Parkinson's disease (PD), possibly caused by alterations in the parasympathetic tonus. Here we explore salivary acetylcholinesterase (AChE) activity as a potential biomarker in PD. Methods. We measured salivary flow, AChE activity, and total protein concentration in 30 PD patients and 49 healthy controls. We also performed exploratory correlation analyses with disease duration, motor symptom severity, autonomic complaints, and other nonmotor symptoms. Results. PD patients displayed significantly decreased salivary flow rate, significantly increased salivary AChE activity, and total protein concentration. Importantly, the AChE activity/total protein ratio was significantly increased in PD patients, suggesting that increased AChE activity cannot be explained solely by upconcentration of saliva. The Unified PD Rating Scale (UPDRS) score displayed significant correlation with total salivary protein (P = 0.002) and near-significant correlation with salivary flow (P = 0.07). Color vision test scores were also significantly correlated with AChE activity (P = 0.04) and total protein levels (P = 0.002). Conclusion. Salivary AChE activity is increased in PD patients compared to healthy controls. Future studies are needed to elucidate whether this parameter reflects the extent of neuronal damage and parasympathetic denervation in the salivary glands of PD patients.

  11. A review on structure-activity relationship of dietary polyphenols inhibiting ?-amylase.

    PubMed

    Xiao, Jianbo; Ni, Xiaoling; Kai, Guoyin; Chen, Xiaoqing

    2013-01-01

    The inhibitory effects of dietary polyphenols against ?-amylase have attracted great interest among researchers. The aim of this review is to give an overview of the research reports on the structure-activity relationship of polyphenols inhibiting ?-amylase. The molecular structures that influence the inhibition are the following: (1) The hydroxylation of flavonoids improved the inhibitory effect on ?-amylase; (2) Presence of an unsaturated 2,3-bond in conjugation with a 4-carbonyl group has been associated with stronger inhibition; (3) The glycosylation of flavonoids decreased the inhibitory effect on ?-amylase depending on the conjugation site and the class of sugar moiety; (4) The methylation and methoxylation of flavonoids obviously weakened the inhibitory effect; (5) The galloylated catechins have higher inhibition than nongalloylated catechins; the catechol-type catechins were stronger than the pyrogallol-type catechins; the inhibition activities of the catechins with 2,3-trans structure were higher than those of the catechins with 2,3-cis structure; (6) Cyanidin-3-glucoside showed higher inhibition against than cyanidin and cyanidin-3-galactoside and cyanidin-3,5-diglucoside had no inhibitory activity; (7) Ellagitannins with ?-galloyl groups at glucose C-1 positions have higher inhibitory effect than the ?-galloyl and nongalloyl compounds and the molecular weight of ellagitannins is not an important element. PMID:23391016

  12. Amylase Test

    MedlinePLUS

    ... obstruction and pancreatic cancers . In general, urine amylase levels rise in proportion to blood amylase levels and will ... medications that I am taking affect the amylase level? ... to rise include aspirin, diuretics , oral contraceptives, corticosteroids, indomethacin, ethyl ...

  13. Salivary alpha amylase diurnal pattern and stress response are associated with body mass index in low-income preschool-aged children.

    PubMed

    Miller, Alison L; Sturza, Julie; Rosenblum, Katherine; Vazquez, Delia M; Kaciroti, Niko; Lumeng, Julie C

    2015-03-01

    Physiological stress responses are proposed as a pathway through which stress can "get under the skin" and lead to health problems, specifically obesity. We tested associations of salivary alpha amylase (sAA) diurnal patterns and stress responses with body mass index (BMI) in young, low-income children (51% male; 54% non-Hispanic white). Diurnal saliva samples were collected three times per day across three days for 269 children (M age 50.8 months, SD 6.3). Individual sAA intercept and slope values were calculated using random effect models to represent morning sAA levels and rate of sAA change across the day. A subset of children (n=195; M age 56.6 months, SD 6.9) participated in a lab-based behavioral stress protocol. Area under the curve increase (AUCI) across four timepoints was calculated to represent increase in sAA output during stress elicitation. Children were weighed and height measured and BMI z-score was calculated. Linear regression was used to evaluate associations of sAA intercept, sAA slope, and sAA AUCI with BMI z-score, controlling for child age, sex, and race/ethnicity; maternal weight status; and family income-to-needs ratio. Diurnal and stress-response sAA patterns were related to child adiposity: for each 1-standard deviation unit (SDU) decrease in morning sAA level, the child's BMI z-score increased by 0.11 (SE 0.05) SDU's (p<.04); for each 1-SDU increase in sAA slope across the day, the child's BMI z-score increased by 0.12 (SE 0.05) SDU's (p<.03); and for each 1-SDU decrease in sAA AUCI during the stress elicitation, the child's BMI z-score increased by 0.14 (SE 0.06) SDU's (p<.03). Blunted stress responses and atypical diurnal patterns of sAA have been found following exposure to chronic life stressors such as poverty. Findings suggest that associations of stress, sAA, and elevated body mass index may develop very early in the lifespan. PMID:25588701

  14. Salivary mental stress proteins.

    PubMed

    Obayashi, Konen

    2013-10-21

    Of the major diagnostic specimen types, saliva is one of the most easily collected. Many studies have focused on the evaluation of salivary proteins secreted by healthy people and patients with various diseases during responses to acute mental stress. In particular, such studies have focused on cortisol, ?-amylase, chromogranin A (CgA), and immunoglobulin A (IgA) as salivary stress markers. Each of these salivary stress markers has its own strengths and weaknesses as well as data gaps related to many factors including collection technique. In this review, we summarize the critical knowledge of the positive and negative attributes and data gaps pertaining to each salivary stress marker. PMID:23939251

  15. Partial characterization of cold active amylases and proteases of Streptomyces sp. from Antarctica

    PubMed Central

    Cotârle?, Mihaela; Negoi??, Teodor Gh.; Bahrim, Gabriela E.; Stougaard, Peter

    2011-01-01

    The aim of this study was to isolate novel enzyme-producing bacteria from vegetation samples from East Antarctica and also to characterize them genetically and biochemically in order to establish their phylogeny. The ability to grow at low temperature and to produce amylases and proteases cold-active was also tested. The results of the 16S rRNA gene sequence analysis showed that the 4 Alga rRNA was 100% identical to the sequences of Streptomyces sp. rRNA from Norway and from the Solomon Islands. The Streptomyces grew well in submerged system at 20°C, cells multiplication up to stationary phase being drastically increased after 120 h of submerged cultivation. The beta-amylase production reached a maximum peak after seven days, while alpha-amylase and proteases were performing biosynthesis after nine days of submerged cultivation at 20°C. Newly Streptomyces were able to produce amylase and proteases in a cold environment. The ability to adapt to low temperature of these enzymes could make them valuable ingredients for detergents, the food industry and bioremediation processes which require low temperatures. PMID:24031702

  16. Grape seed and tea extracts and catechin 3-gallates are potent inhibitors of ?-amylase and ?-glucosidase activity.

    PubMed

    Yilmazer-Musa, Meltem; Griffith, Anneke M; Michels, Alexander J; Schneider, Erik; Frei, Balz

    2012-09-12

    This study evaluated the inhibitory effects of plant-based extracts (grape seed, green tea, and white tea) and their constituent flavan-3-ol monomers (catechins) on ?-amylase and ?-glucosidase activity, two key glucosidases required for starch digestion in humans. To evaluate the relative potency of extracts and catechins, their concentrations required for 50 and 90% inhibition of enzyme activity were determined and compared to the widely used pharmacological glucosidase inhibitor, acarbose. Maximum enzyme inhibition was used to assess relative inhibitory efficacy. Results showed that grape seed extract strongly inhibited both ?-amylase and ?-glucosidase activity, with equal and much higher potency, respectively, than acarbose. Whereas tea extracts and catechin 3-gallates were less effective inhibitors of ?-amylase, they were potent inhibitors of ?-glucosidase. Nongallated catechins were ineffective. The data show that plant extracts containing catechin 3-gallates, in particular epigallocatechin gallate, are potent inhibitors of ?-glucosidase activity and suggest that procyanidins in grape seed extract strongly inhibit ?-amylase activity. PMID:22697360

  17. Studies on activity, distribution, and zymogram of protease, ?-amylase, and lipase in the paddlefish Polyodon spathula.

    PubMed

    Ji, H; Sun, H T; Xiong, D M

    2012-06-01

    A series of biochemical determination and electrophoretic observations have been conducted to analyze the activities and characteristics of protease, ?-amylase, and lipase of paddlefish Polyodon spathula. The results obtained have been compared with those of bighead carp (Aristichthys nobilis) and hybrid sturgeon (Huso dauricus ? × Acipenser schrenki Brandt ?), in order to increase available knowledge of the physiological characteristics of this sturgeon species and to gain information with regard to its nutrition. Further, a comparative study of enzymatic activity, distribution, and characterization between commercial feed-reared paddlefish (CG) and natural live food-reared (NG) paddlefish was conducted. Results showed that higher proteolytic activity was observed in the pH range 2.5-3.0 and at a pH of 7.0 for paddlefish. Levels of acid protease activity of paddlefish were similar to that of hybrid sturgeon, and significantly higher than that of bighead carp. The inhibition assay of paddlefish showed that the rate of inhibition of tosyl-phenylalanine chloromethyl ketone was approximately 2.6-fold that of tosyl-lysine chloromethyl ketone. There was no significant difference observed for acid protease activity between PG and CG groups, whereas the activity of alkaline protease, ?-amylase, and lipase in the PG group were significantly lower than those in the CG group. The substrate sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis further showed that there were certain types of enzymes, especially ?-amylase, with similar molecular mass in the paddlefish and hybrid sturgeon. It can be inferred that acid digestion was main mechanism for protein hydrolysis in paddlefish, as reported for other fishes with a stomach. This indicates that the paddlefish requires higher alkaline protease, ?-amylase, and lipase activity to digest natural live food. PMID:21894570

  18. Phenotypic Variation for Diastatic power, ß-Amylase Activity, and ß-Amylase Thermostability vs. Allelic Variation at the Bmy1 Locus in a Sample of North American Barley Germplasm

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Malting quality data including diastatic power, ß-amylase activity, and ß-amylase thermostability, were collected on malts from three barley (Hordeum vulgare L.) breeding program trials containing two growth habits and 165 lines grown in multiple environments. We attempted to identify causal polymor...

  19. Comparison of alpha-amylase activity in larval stages of flour beetles, Tribolium confusum (Coleoptera: Tenebionidae).

    PubMed

    Bandani, A R; Balvasi, A

    2006-01-01

    Flour beetles attack stored grain products such as flour, cereals, meal, dried pet food, dried flowers and even dried museum specimens and other foods in the house. Stored-product insects cause tremendous losses by lowering weight, germination rate, nutritional value and grain grade. These beetles are of the most important pests of stored products in the home and grocery stores. The adult female may live for as long as two years, depositing 300 to 400 eggs. The life cycle requires one to four months when temperatures are favorable. Several methods could be used to control this insect including synthetic insecticides, biological control, physical control and transgenic plant carrying gene of interest. Chemical controls are discouraged due to pesticide residue in the commodities and resistance in insects. The study of insect digestive enzymes seems to make sense in the realization that the gut is the major interface between the insect and its environment. Hence, an understanding of digestive enzyme function is essential when developing methods of insect control such as the use of enzyme inhibitors and transgenic plants to control insect pests. Therefore, the aim of the current study was to get a good understanding from enzyme composition of different larval stages of the insect and finally characterize amylase which is the key enzyme in digestive system of this insect. For alpha-amylase study whole larvae were homogenized in 0.02 M phosphate buffer at pH 7.2. The homogenates were separately transferred to a 1.5 ml of centrifuge tubes and centrifuged at 15000xg for 20 min at 4degrees C. The supernatants were used as enzyme source in assays. alpha-Amylase activity was assayed by the dinitrosalicylic acid (DNS) procedure using 1% soluble starch (Merck) as substrate. The results show that enzyme activity (OD) in the first, second, third and fourth larval stages were 0.5, 1.15, 1.35 and 1.362, respectively. There are significant differences in amylase activity in different larval stages; however, there are no significant differences in the enzyme activity of two last larval stages. Wheat grain is a major source of starch and insect that feed on the wheat grain or flour made from wheat rely heavily on their amylase for starch hydrolyze and this could be the main reason that larval stages even first larval instar has such a high amount of alpha-amylase. PMID:17385521

  20. Tracking amylolytic enzyme activities during congress mashing with North American barley cultivars: Comparisons of patterns of activity and ß-amylases with differing Bmy1 ...correlations of amylolytic enzyme activities

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was conducted to test three hypotheses: 1) that a-amylase will have less consistent patterns of activity during mashing than ß-amylase and limit dextrinase 2) that differing ß-amylase 1 intron III alleles (Bmy1.a and Bmy1.b) would not be useful in predicting high or low activities or th...

  1. Hotspot activating PRKD1 somatic mutations in polymorphous low-grade adenocarcinomas of the salivary glands.

    PubMed

    Weinreb, Ilan; Piscuoglio, Salvatore; Martelotto, Luciano G; Waggott, Daryl; Ng, Charlotte K Y; Perez-Ordonez, Bayardo; Harding, Nicholas J; Alfaro, Javier; Chu, Kenneth C; Viale, Agnes; Fusco, Nicola; da Cruz Paula, Arnaud; Marchio, Caterina; Sakr, Rita A; Lim, Raymond; Thompson, Lester D R; Chiosea, Simion I; Seethala, Raja R; Skalova, Alena; Stelow, Edward B; Fonseca, Isabel; Assaad, Adel; How, Christine; Wang, Jianxin; de Borja, Richard; Chan-Seng-Yue, Michelle; Howlett, Christopher J; Nichols, Anthony C; Wen, Y Hannah; Katabi, Nora; Buchner, Nicholas; Mullen, Laura; Kislinger, Thomas; Wouters, Bradly G; Liu, Fei-Fei; Norton, Larry; McPherson, John D; Rubin, Brian P; Clarke, Blaise A; Weigelt, Britta; Boutros, Paul C; Reis-Filho, Jorge S

    2014-11-01

    Polymorphous low-grade adenocarcinoma (PLGA) is the second most frequent type of malignant tumor of the minor salivary glands. We identified PRKD1 hotspot mutations encoding p.Glu710Asp in 72.9% of PLGAs but not in other salivary gland tumors. Functional studies demonstrated that this kinase-activating alteration likely constitutes a driver of PLGA. PMID:25240283

  2. Employing in vitro directed molecular evolution for the selection of ?-amylase variant inhibitors with activity toward cotton boll weevil enzyme.

    PubMed

    da Silva, Maria Cristina Mattar; Del Sarto, Rafael Perseghini; Lucena, Wagner Alexandre; Rigden, Daniel John; Teixeira, Fabíola Rodrigues; Bezerra, Caroline de Andrade; Albuquerque, Erika Valéria Saliba; Grossi-de-Sa, Maria Fatima

    2013-09-20

    Numerous species of insect pests attack cotton plants, out of which the cotton boll weevil (Anthonomus grandis) is the main insect in Brazil and must be controlled to avert large economic losses. Like other insect pests, A. grandis secretes a high level of ?-amylases in the midgut lumen, which are required for digestion of carbohydrates. Thus, ?-amylase inhibitors (?-AIs) represent a powerful tool to apply in the control of insect pests. Here, we applied DNA shuffling and phage display techniques and obtained a combinatorial library containing 10? ?-AI variant forms. From this library, variants were selected exhibiting in vitro affinity for cotton boll weevil ?-amylases. Twenty-six variant sequences were cloned into plant expression vectors and expressed in Arabidopsis thaliana. Transformed plant extracts were assayed in vitro to select specific and potent ?-amylase inhibitors against boll weevil amylases. While the wild type inhibitors, used to create the shuffled library, did not inhibit the A. grandis ?-amylases, three ?-AI mutants, named ?-AIC3, ?-AIA11 and ?-AIG4 revealed high inhibitory activities against A. grandis ?-amylases in an in vitro assay. In summary, data reported here shown the potential biotechnology of new ?-AI variant genes for cotton boll weevil control. PMID:23892157

  3. alpha-Amylase inhibitory activity of some Malaysian plants used to treat diabetes; with particular reference to Phyllanthus amarus.

    PubMed

    Ali, Hasenah; Houghton, P J; Soumyanath, Amala

    2006-10-11

    Extracts of six selected Malaysian plants with a reputation of usefulness in treating diabetes were examined for alpha-amylase inhibition using an in vitro model. Inhibitory activity studied by two different protocols (with and without pre-incubation) showed that Phyllanthus amarus hexane extract had alpha-amylase inhibitory properties. Hexane and dichloromethane extracts of Anacardium occidentale, Lagerstroemia speciosa, Averrhoa bilimbiPithecellobium jiringa and Parkia speciosa were not active when tested without pre-incubation. Extraction and fractionation of Phyllanthus amarus hexane extract led to the isolation of dotriacontanyl docosanoate, triacontanol and a mixture of oleanolic acid and ursolic acid. Dotriacontanyl docosanoate and the mixture of oleanolic acid and ursolic acid are reported from this plant species for the first time. All compounds were tested in the alpha-amylase inhibition assay and the results revealed that the oleanolic acid and ursolic acid (2:1) mixture was a potent alpha-amylase inhibitor with IC(50)=2.01 microg/ml (4.41 microM) and that it contributes significantly to the alpha-amylase inhibition activity of the extract. Three pure pentacyclic triterpenoids, oleanolic acid, ursolic acid and lupeol were shown to inhibit alpha-amylase. PMID:16678367

  4. ?-Amylase sensor based on the degradation of oligosaccharide hydrogel films monitored with a quartz crystal sensor.

    PubMed

    Gibbs, Martin John; Biela, Anna; Krause, Steffi

    2015-05-15

    ?-Amylase hydrolyses starch molecules to produce smaller oligosaccharides and sugars. Amylases are of great importance in biotechnology and find application in fermentation, detergents, food and the paper industry. The measurement of ?-amylase activity in serum and urine has been used in the diagnosis of acute pancreatitis. Salivary amylase has also been shown to be a stress indicator. Sensor coatings suitable for the detection of ?-amylase activity have been developed. Oligosaccharides such as glycogen and amylopectin were spin-coated onto gold coated quartz crystals with a base frequency of 10 MHz. The films were subsequently cross-linked with hexamethylene diisocyanate. Film degradation was monitored with a quartz crystal microbalance (QCM) and electrochemical impedance measurements. The films were shown to be stable in phosphate buffered saline (PBS). Addition of ?-amylase to the solution resulted in the rapid degradation of the films. The maximum rate of degradation was found to be strongly dependent on the amylase activity in the range typically found in serum when diagnosing pancreatitis (0.08-8 U/ml). Sensor responses in serum were found to be very similar to those obtained in buffer indicating the absence of non-specific binding. PMID:25266253

  5. Evaluation of Ten Wild Nigerian Mushrooms for Amylase and Cellulase Activities

    PubMed Central

    Adeoyo, Olusegun Richard

    2011-01-01

    Amylases and cellulases are important enzymes that can be utilized for various biological activities. Ten different wild Nigerian mushrooms (Agaricus blazei, Agaricus sp., Corilopsis occidentalis, Coriolus versicolor, Termitomyces clypeatus, Termitomyces globulus, Pleurotus tuber-regium, Podoscypha bolleana, Pogonomyces hydnoides, and Nothopanus hygrophanus) were assayed for production of these secondary metabolites. The results revealed that most of the tested wild fungi demonstrated very good amylase and cellulase activities. With the incorporation of carboxymethyl-cellulose (a carbon source) into the culture medium, Agaricus blazei had the highest amylolytic activity of 0.60 unit/mL (at 25?, pH 6.8). This was followed in order by P. tuber-regium and Agaricus sp. with 0.42 and 0.39 unit/mL, respectively (p ? 0.05). Maltose and sucrose supplementation into the submerged liquid medium made N. hygrophanus and P. hydnoides to exhibit very low amylase activities of 0.09 and 0.11 unit/mL, respectively. Introducing peptone (an organic nitrogen source) into the basal medium enhanced the ability of C. versicolor to produce a cellulase value of 0.74 unit/mL. Other organic nitrogen sources that supported good cellulase activities were yeast extract and urea. Sodium nitrate (inorganic nitrogen source) generally inhibited cellulase production in all mushrooms. The best carbon source was carboxymethyl-cellulose, which promoted very high cellulase activity of 0.67 unit/mL in C. versicolor, which was followed in order by P. tuber-regium, T. chypeatus, and C. occidentalis (p ? 0.05). Sucrose was the poorest carbon compound, supporting the lowest values of 0.01, 0.01, and 0.14 unit/mL in P. hydnoides, A. blazei, and Agaricus sp., respectively. PMID:22783085

  6. Salivary Gland Secretion.

    ERIC Educational Resources Information Center

    Dorman, H. L.; And Others

    1981-01-01

    Describes materials and procedures for an experiment utilizing a live dog to demonstrate: (1) physiology of the salivary gland; (2) parasympathetic control of the salivary gland; (3) influence of varying salivary flow rates on sodium and potassium ions, osmolarity and pH; and (4) salivary secretion as an active process. (DS)

  7. Chloride Activated Halophilic ?-Amylase from Marinobacter sp. EMB8: Production Optimization and Nanoimmobilization for Efficient Starch Hydrolysis

    PubMed Central

    Kumar, Sumit; Khare, S. K.

    2015-01-01

    Halophiles have been perceived as potential source of novel enzymes in recent years. The interest emanates from their ability to catalyze efficiently under high salt and organic solvents. Present work encompasses production optimization and nanoimmobilization of an ?-amylase from moderately halophilic Marinobacter sp. EMB8. Media ingredients and culture conditions were optimized by “one-at-a-time approach.” Starch was found to be the best carbon source at 5% (w/v) concentration. Glucose acted as catabolic repressor for amylase production. Salt proved critical for amylase production and maximum production was attained at 5% (w/v) NaCl. Optimization of various culture parameters resulted in 48.0?IU/mL amylase production, a 12-fold increase over that of unoptimized condition (4.0?IU/mL). ?-Amylase was immobilized on 3-aminopropyl functionalized silica nanoparticles using glutaraldehyde as cross-linking agent. Optimization of various parameters resulted in 96% immobilization efficiency. Starch hydrolyzing efficiency of immobilized enzyme was comparatively better. Immobilized ?-amylase retained 75% of its activity after 5th cycle of repeated use. PMID:25667773

  8. Effect of ionic liquids on the structure, stability and activity of two related ?-amylases.

    PubMed

    Dabirmanesh, Bahareh; Daneshjou, Sara; Sepahi, Abbas Akhavan; Ranjbar, Bijan; Khavari-Nejad, Ramazan Ali; Gill, Pooria; Heydari, Akbar; Khajeh, Khosro

    2011-01-01

    Ionic liquids are recognized as green solvents for carbohydrates dissolution. However, only a limited number of studies have been carried out to investigate their effect on carbohydrate hydrolyzing enzymes. We have investigated the influence of two water miscible ionic liquids on the activity, stability and structure of two related ?-amylases from Bacillus amyloliquefaciens and Bacillus lichiniformis. Upon changes in ionic liquids concentrations, both enzymes activity and stability were reduced. Associated thermodynamic and conformational changes were observed using differential scanning calorimetry and fluorescence techniques. Thermal denaturation was accompanied by aggregation in both aqueous buffer and [BMIm][Cl] but [HMIm][Cl] significantly suppressed aggregation. PMID:20946913

  9. Different Proportions of Huangqi (Radix Astragali Mongolici) and Honghua (Flos Carthami) Injection on ?-Glucosidase and ?-Amylase Activities

    PubMed Central

    Banbury, Linda

    2015-01-01

    Objective. To study the effect of different proportions of Huangqi (Radix Astragali Mongolici) and Honghua (Flos Carthami) injection on ?-glucosidase and ?-amylase activity simultaneously. Methods. The injections were prepared according to the standards of the China Food and Drug Administration. The assay for potential ?-glucosidase inhibitors was based on the hydrolysis of 4-methylumbelliferyl-?-D-glucopyranoside (4-MUG). The ?-amylase EnzChek assay kit was used to determine potential ?-amylase inhibitors. Acarbose was the positive control. Results. The half maximal (50%) inhibitory concentration (IC50) of acarbose against ?-glucosidase and ?-amylase was (1.8 ± 0.4) ?g/mL and (227 ± 32) ?g/mL, respectively. Honghua showed significant inhibition of ?-glucosidase activity compared with Huangqi (P < 0.01). Honghua inhibited ?-amylase activity, but Huangqi did not. IC50s for ?-glucosidase inhibition by mixtures at 10?:?1, 5?:?1, and 2?:?1 were significantly lower than those at the 20?:?1 mixture (P < 0.01). ?-Amylase inhibition by the 2?:?1 mixture was significantly higher than that by the 20?:?1, 10?:?1, and 5?:?1 mixtures at 500??g/mL and 1000??g/mL (P < 0.01), with 5?:?1 significantly higher than 20?:?1 and 10?:?1 at 1000??g/mL (P < 0.01). Conclusion. Honghua significantly inhibited ?-glucosidase activity compared with Huangqi (P < 0.01). For simultaneous inhibition of ?-glucosidase and ?-amylase activities, the mixtures at 2?:?1 and 5?:?1 exhibited significant effects compared with those at 20?:?1 (P < 0.01).

  10. MDM2 Is Required for Suppression of Apoptosis by Activated Akt1 in Salivary Acinar Cells?

    PubMed Central

    Limesand, Kirsten H.; Schwertfeger, Kathryn L.; Anderson, Steven M.

    2006-01-01

    Chronic damage to the salivary glands is a common side effect following head and neck irradiation. It is hypothesized that irreversible damage to the salivary glands occurs immediately after radiation; however, previous studies with rat models have not shown a causal role for apoptosis in radiation-induced injury. We report that etoposide and gamma irradiation induce apoptosis of salivary acinar cells from FVB control mice in vitro and in vivo; however, apoptosis is reduced in transgenic mice expressing a constitutively activated mutant of Akt1 (myr-Akt1). Expression of myr-Akt1 in the salivary glands results in a significant reduction in phosphorylation of p53 at serine18, total p53 protein accumulation, and p21WAF1 or Bax mRNA following etoposide or gamma irradiation of primary salivary acinar cells. The reduced level of p53 protein in myr-Akt1 salivary glands corresponds with an increase in MDM2 phosphorylation in vivo, suggesting that the Akt/MDM2/p53 pathway is responsible for suppression of apoptosis. Dominant-negative Akt blocked phosphorylation of MDM2 in salivary acinar cells from myr-Akt1 transgenic mice. Reduction of MDM2 levels in myr-Akt1 primary salivary acinar cells with small interfering RNA increases the levels of p53 protein and renders these cells susceptible to etoposide-induced apoptosis in spite of the presence of activated Akt1. These results indicate that MDM2 is a critical substrate of activated Akt1 in the suppression of p53-dependent apoptosis in vivo. PMID:16982679

  11. Variation in ?-amylase activity and thermostability in Tibetan annual wild and cultivated barley genotypes*

    PubMed Central

    Zhang, Hai-tao; Chen, Tian-long; Zhang, Bing-lin; Wu, De-zhi; Huang, Ye-chang; Wu, Fei-bo; Zhang, Guo-ping

    2014-01-01

    ?-Amylase activity (BAA) and thermostability (BAT) are important traits for malt quality. In this study, 138 Tibetan annual wild barley accessions and 20 cultivated genotypes differing in BAA were planted and analyzed in 2009 and 2012. Significant differences were detected among genotypes in BAA and BAT. The cultivated genotypes had a mean BAA of 1137.6 U/g and a range of from 602.1 to 1407.5 U/g, while the wild accessions had a mean of 1517.9 U/g and a range of from 829.7 to 2310.0 U/g. The cultivated genotypes had a mean relative residual ?-amylase activity (RRBAA) of 61.6% and a range of from 22.2% to 82.3%, while the wild barleys had a mean of 57.8% and a range of from 21.9% to 96.1%. Moreover, there was a significant difference among genotypes in the response of RRBAA to the temperature and duration of heat treatment. The wild barleys had wider variation in BAA and BAT than cultivated genotypes. PMID:25183034

  12. Enhanced antifungal and insect ?-amylase inhibitory activities of Alpha-TvD1, a peptide variant of Tephrosia villosa defensin (TvD1) generated through in vitro mutagenesis.

    PubMed

    Vijayan, S; Imani, J; Tanneeru, K; Guruprasad, L; Kogel, K H; Kirti, P B

    2012-02-01

    TvD1 is a small, cationic, and highly stable defensin from the weedy legume, Tephrosia villosa with demonstrated in vitro antifungal activity. We show here peptide modifications in TvD1 that lead to enhanced antifungal activities. Three peptide variants, S32R, D37R, and Alpha-TvD1 (-G-M-T-R-T-) with variations in and around the ?2-?3 loop region that imposes the two ?-strands, ?2 and ?3 were generated through in vitro mutagenesis. Alpha-TvD1 exhibited enhanced antifungal activity against the fungal pathogens, Fusarium culmorum and Fusarium oxysporum with respective IC(50) values of 2.5 ?M and 3.0 ?M, when compared to S32R (<5.0 ?M and >5.0 ?M), D37R (5.5 ?M and 4.5 ?M), and the wild type TvD1 (6.5 ?M). Because of the enhanced antifungal activity, this variant peptide was characterized further. Growth of F. culmorum in the presence of Alpha-TvD1 showed deformities in hyphal walls and nuclear damage. With respect to the plant pathogenic bacterium, Pseudomonas syringae pv. tomato strain DC3000, both Alpha-TvD1 and the wild type TvD1 showed comparable antibacterial activity. Both wild type TvD1 and Alpha-TvD1 displayed inhibitory activity against the ?-amylase of the mealworm beetle, Tenebrio molitor (TMA) with the latter showing enhanced activity. The human salivary as well as barley ?-amylase activities were not inhibited even at concentrations of up to 50 ?M, which has been predicted to be due to differences in the pocket size and the size of the interacting loops. Present study shows that the variant Alpha-TvD1 exhibits enhanced antifungal as well as insect ?-amylase inhibitory activity. PMID:22244814

  13. Introducing transglycosylation activity in Bacillus licheniformis ?-amylase by replacement of His235 with Glu.

    PubMed

    Tran, Phuong Lan; Cha, Hyun-Ju; Lee, Jin-Sil; Park, Sung-Hoon; Woo, Eui-Jeon; Park, Kwan-Hwa

    2014-09-01

    To understand the role of His and Glu in the catalytic activity of Bacillus licheniformis ?-amylase (BLA), His235 was replaced with Glu. The mutant enzyme, H235E, was characterized in terms of its mode of action using labeled and unlabeled maltooctaose (Glc8). H235E predominantly produced maltotridecaose (Glc13) from Glc8, exhibiting high substrate transglycosylation activity, with Km=0.38mM and kcat/Km=20.58mM(-1)s(-1) for hydrolysis, and Km2=18.38mM and kcat2/Km2=2.57mM(-1)s(-1) for transglycosylation, while the wild-type BLA exhibited high hydrolysis activity exclusively. Glu235-located on a wide open groove near subsite +1-is likely involved in transglycosylation via formation of an ?-1,4-glycosidic linkage and may recognize and stabilize the non-reducing end glucose of the acceptor molecule. PMID:25117441

  14. Human ?-amylase present in lower-genital-tract mucosal fluid processes glycogen to support vaginal colonization by Lactobacillus.

    PubMed

    Spear, Gregory T; French, Audrey L; Gilbert, Douglas; Zariffard, M Reza; Mirmonsef, Paria; Sullivan, Thomas H; Spear, William W; Landay, Alan; Micci, Sandra; Lee, Byung-Hoo; Hamaker, Bruce R

    2014-10-01

    Lactobacillus colonization of the lower female genital tract provides protection from the acquisition of sexually transmitted diseases, including human immunodeficiency virus, and from adverse pregnancy outcomes. While glycogen in vaginal epithelium is thought to support Lactobacillus colonization in vivo, many Lactobacillus isolates cannot utilize glycogen in vitro. This study investigated how glycogen could be utilized by vaginal lactobacilli in the genital tract. Several Lactobacillus isolates were confirmed to not grow in glycogen, but did grow in glycogen-breakdown products, including maltose, maltotriose, maltopentaose, maltodextrins, and glycogen treated with salivary ?-amylase. A temperature-dependent glycogen-degrading activity was detected in genital fluids that correlated with levels of ?-amylase. Treatment of glycogen with genital fluids resulted in production of maltose, maltotriose, and maltotetraose, the major products of ?-amylase digestion. These studies show that human ?-amylase is present in the female lower genital tract and elucidates how epithelial glycogen can support Lactobacillus colonization in the genital tract. PMID:24737800

  15. Comparisons of amylolytic enzyme activities and ß-amylases with differing Bmy1 intron III alleles to sugar production during congress mashing with North American barley cultivars

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was conducted to determine the relationships between patterns of activity development of malt amylolytic enzymes (a-amylase, ß-amylase, and limit dextrinase) and sugar production in two- and six-row North American cultivars during the course of Congress mashing and to test two hypotheses:...

  16. Utilization of Different Bmy1 Intron III Alleles for Predicting ß-Amylase Activity and Thermostability in Wild and Cultivated Barley

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Polymorphisms in intron III of barley (Hordeum vulgare L.) endosperm-specific beta-amylase (Bmy1) have been associated with beta-amylase activity and thermostability and are thought to have potential as a selective marker for breeding elite malting cultivars. The third intron of Bmy1 was sequenced ...

  17. Differential RNA Expression of Bmy1 During Late Seed Development in Wild and Cultivated Barley and the Association With ß-Amylase Activity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Four genotypes carrying different ß-amylase 1 (Bmy1) intron III alleles (Bmy1.a, Bmy1.b, Bmy1.c, and Bmy1.d) were analyzed for differences in Bmy1 DNA sequence, Bmy1 RNA expression, ß-amylase activity and protein, and total protein during late seed development. Wild barleys Ashqelon (Bmy1.c) and PI...

  18. Glucan, Water Dikinase Activity Stimulates Breakdown of Starch Granules by Plastidial ?-Amylases1[W][OA

    PubMed Central

    Edner, Christoph; Li, Jing; Albrecht, Tanja; Mahlow, Sebastian; Hejazi, Mahdi; Hussain, Hasnain; Kaplan, Fatma; Guy, Charles; Smith, Steven M.; Steup, Martin; Ritte, Gerhard

    2007-01-01

    Glucan phosphorylating enzymes are required for normal mobilization of starch in leaves of Arabidopsis (Arabidopsis thaliana) and potato (Solanum tuberosum), but mechanisms underlying this dependency are unknown. Using two different activity assays, we aimed to identify starch degrading enzymes from Arabidopsis, whose activity is affected by glucan phosphorylation. Breakdown of granular starch by a protein fraction purified from leaf extracts increased approximately 2-fold if the granules were simultaneously phosphorylated by recombinant potato glucan, water dikinase (GWD). Using matrix-assisted laser-desorption ionization mass spectrometry several putative starch-related enzymes were identified in this fraction, among them ?-AMYLASE1 (BAM1; At3g23920) and ISOAMYLASE3 (ISA3; At4g09020). Experiments using purified recombinant enzymes showed that BAM1 activity with granules similarly increased under conditions of simultaneous starch phosphorylation. Purified recombinant potato ISA3 (StISA3) did not attack the granular starch significantly with or without glucan phosphorylation. However, starch breakdown by a mixture of BAM1 and StISA3 was 2 times higher than that by BAM1 alone and was further enhanced in the presence of GWD and ATP. Similar to BAM1, maltose release from granular starch by purified recombinant BAM3 (At4g17090), another plastid-localized ?-amylase isoform, increased 2- to 3-fold if the granules were simultaneously phosphorylated by GWD. BAM activity in turn strongly stimulated the GWD-catalyzed phosphorylation. The interdependence between the activities of GWD and BAMs offers an explanation for the severe starch excess phenotype of GWD-deficient mutants. PMID:17631522

  19. Concurrent Transient Activation of Wnt/{beta}-Catenin Pathway Prevents Radiation Damage to Salivary Glands

    SciTech Connect

    Hai Bo; Yang Zhenhua; Shangguan Lei; Zhao Yanqiu [Institute for Regenerative Medicine, Scott and White Hospital, Molecular and Cellular Medicine Department, Texas A and M Health Science Center, Temple, Texas (United States); Boyer, Arthur [Department of Radiology, Scott and White Hospital, Temple, Texas (United States); Liu, Fei, E-mail: fliu@medicine.tamhsc.edu [Institute for Regenerative Medicine, Scott and White Hospital, Molecular and Cellular Medicine Department, Texas A and M Health Science Center, Temple, Texas (United States)

    2012-05-01

    Purpose: Many head and neck cancer survivors treated with radiotherapy suffer from permanent impairment of their salivary gland function, for which few effective prevention or treatment options are available. This study explored the potential of transient activation of Wnt/{beta}-catenin signaling in preventing radiation damage to salivary glands in a preclinical model. Methods and Materials: Wnt reporter transgenic mice were exposed to 15 Gy single-dose radiation in the head and neck area to evaluate the effects of radiation on Wnt activity in salivary glands. Transient Wnt1 overexpression in basal epithelia was induced in inducible Wnt1 transgenic mice before together with, after, or without local radiation, and then saliva flow rate, histology, apoptosis, proliferation, stem cell activity, and mRNA expression were evaluated. Results: Radiation damage did not significantly affect activity of Wnt/{beta}-catenin pathway as physical damage did. Transient expression of Wnt1 in basal epithelia significantly activated the Wnt/{beta}-catenin pathway in submandibular glands of male mice but not in those of females. Concurrent transient activation of the Wnt pathway prevented chronic salivary gland dysfunction following radiation by suppressing apoptosis and preserving functional salivary stem/progenitor cells. In contrast, Wnt activation 3 days before or after irradiation did not show significant beneficial effects, mainly due to failure to inhibit acute apoptosis after radiation. Excessive Wnt activation before radiation failed to inhibit apoptosis, likely due to extensive induction of mitosis and up-regulation of proapoptosis gene PUMA while that after radiation might miss the critical treatment window. Conclusion: These results suggest that concurrent transient activation of the Wnt/{beta}-catenin pathway could prevent radiation-induced salivary gland dysfunction.

  20. Antioxidant and ?-amylase inhibitory activities of extract and isolates from Zygogynum pancheri subsp. arrhantum.

    PubMed

    Keskes, Henda; Litaudon, Marc; Cherif, Atef; Belhadj, Sahla; Hamdi, Besma; El Feki, Abdelfattah; Dumontet, Vincent; Ben Salah, Abdelhamid; Damak, Mohamed; Allouche, Noureddine

    2014-12-01

    One new sesquiterpenoid (5R(*),8R(*),9R(*),10R(*))-cinnamolide (8), and seven known compounds, 5-hydroxy-7-methoxyflavonone (1), 8-hydroxy-3-(4'-hydroxyphenyl)-6,7-(2?,2?-dimethylchromene)-tetralone (2), 8-hydroxy-3-(3',4'-dihydroxyphenyl)-6,7-(2?,2?-dimethylchromene)-tetralone (3), 1?-E-O-p-methoxycinnamoyl-bemadienolide (4), 1?-O-(E-cinnamoyl)-6?-hydroxy-9-epi-polygodial (5), 1?-O-(E-cinnamoyl)-6?-hydroxypolygodial (6), and 1?-O-E-cinnamoylpolygodial (7) were isolated from the ethyl acetate extract of barks of Zygogynum pancheri subsp. arrhantum (Winteraceae). The structures of these molecules were assigned predominantly based on spectral data. The structure of compound 8 was confirmed by X-ray crystallographic analysis. Compounds 2 and 3 exhibited significant antioxidant activity, whereas compounds 1 and 4-7 showed significant ?-amylase inhibitory activity. PMID:25034255

  1. Effect of Oral Administration of Intact Casein on Gastrointestinal Hormone Secretion and Pancreatic ?-Amylase Activity in Korean Native Steer

    PubMed Central

    Lee, S. B.; Choi, C. W.; Jin, Y. C.; Wang, T.; Lee, K. H.; Ku, M. B.; Hwang, J. H.; Kim, K. H.; Vega, R. S. A.; Lee, H. G.

    2013-01-01

    Three Korean native steers (779±24 kg) fitted with duodenal cannulas were used in a 3×3 Latin square design to investigate the influence of oral administration of soluble proteins, intact casein (IC) and acid hydrolyzed casein (AHC), on gastrointestinal hormone (GIH) secretion in the blood and pancreatic ?-amylase activity in the duodenum. Oral treatment consisted of a basic diet (control), IC (C+100% protein), or AHC (C+80% amino acid, 20% peptide) for 21 d. Blood and duodenum samples were collected for measurement of serum GI hormones, and pancreatic ?-amylase activity was determined at 900, 1030, 1330, 1630, and 1930 h after feeding on d 21 of treatment. The levels of serum cholecystokinin (CCK) and secretin in the IC treatment group were higher compared to the other treatment groups (p<0.05). In addition to the changes in CCK and secretin levels upon IC treatment, the pancreatic ?-amylase activity in the duodenum was higher in the IC group compared to the control diet group (p<0.05). The response of serum ghrelin to IC and AHC treatment was in accordance with the response of serum secretin. The level of peptide fragments flowing in the duodenum was higher in the IC treatment group than the other treatment groups (p<0.05). In conclusion, this study demonstrated that an increase in duodenal CCK and secretin upon IC oral administration increased pancreatic ?-amylase secretion. In addition, ghrelin may be associated with GI hormone secretion in Korean native steers. PMID:25049835

  2. Salivary enzymes in peptic ulcer disease

    PubMed Central

    Motamedi, Mojdeh; Mansour-Ghanaei, Fariborz; Sariri, Reyhaneh; Vesal, Mahmoud

    2013-01-01

    Aim Peptic ulcer, the common disease of the upper gastro-intestinal tract, occurs in about 5–10% of the world's population. Therefore, diagnosis of trace disease progression with a noninvasive method is of prime importance in the field of healthcare research. The aim of this study was to evaluate the validity of salivary enzymes as noninvasive biomarkers for peptic ulcer. Materials and methods In practice, 34 peptic ulcer patients and 30 healthy subjects donated their un-stimulated saliva samples after 8 h of fasting. The activity of some selected enzymes was measured using appropriate enzymatic assay methods. Results The results indicated an overall alternation in enzymatic activity of saliva in patients suffering from peptic ulcer. Biological activity of a-amylase, peroxidase and lactate dehydrogenase, showed significantly higher values in almost all patients as compared to control subjects. Conclusions Based on the results of salivary enzyme activity, it was concluded that besides the influence of their peptic ulcer on enzyme activity of saliva, the considerably higher activity of a-amylase could also be related to the major role of the enzyme on physiological oxidative stress. PMID:25737890

  3. Salivary parameters of relevance for assessing caries activity in individuals and populations.

    PubMed

    Tenovuo, J

    1997-02-01

    A review of the non-microbial salivary parameters with respect to their possible association with caries activity is presented. The parameters are limited to those which already are or at least in the near future will obviously be simple enough, also for clinical purposes. Salivary flow rate is undoubtedly the most important single parameter since the cariostatic activity or efficacy of practically all other salivary parameters depends on the flow rate. Flow rate as such has no linear association with dental caries but there seems to exist an individual "threshold" limit which is decisive for enhanced caries activity. This threshold limit varies among different individuals and therefore the so-called normal values for unstimulated or stimulated flow rate are more reliable on a population level than among individuals for screening purposes. In any individual a regular and longitudinal follow-up of the flow rate is of higher clinical value than only a single cross-sectional measurement. Salivary buffer effect has only a weak negative association with caries activity and again, this effect is of greater clinical significance on a population level. Since the decisive processes in caries attack occur within or under the dental plaque, the buffering effect of saliva is limited and obviously more important to screen for erosion-than caries-prone individuals. Although important for dental health, none of the salivary antimicrobial agents as such has shown any strong association with caries activity. The only ones with some evidence of a regulatory role are secretory IgA antibodies, hypothiocyanite ions, and agglutinins. However, the data are controversial and it seems that instead of measuring individual parameters, the assessment of saliva's functional properties (such as the ability to aggregate bacteria, prevent their adhesion to hydroxyapatite or sugar metabolism etc.) is more important for clinical purposes. Of the parameters involved in de- and remineralization process, only salivary fluoride content has some association with caries susceptibility but its diagnostic or predictive value is questionable. PMID:9088696

  4. Patterns of puffing activity in the salivary gland chromosomes of Drosophila

    Microsoft Academic Search

    Michael Ashburner

    1969-01-01

    The autosomal salivary gland chromosome puffing patterns of Drosophila simulans are described and compared with the puffing patterns of the sibling species D. melanogaster. During the late third larval instar and the prepupal period the patterns of puffing activity of these two species are similar — approximately 50% of the puffs common to both species showing identical activities. The remaining

  5. Patterns of puffing activity in the salivary gland chromosomes of Drosophila

    Microsoft Academic Search

    Michael Ashburner

    1967-01-01

    The patterns of puffing activity have been studied during the late larval and prepupal stages of Drosophila melanogaster. On the major salivary gland autosomes (chromosomes 2 and 3) 108 loci form puffs at some time during these developmental stages. The timing and pattern of activity of 83 of these puffs is found to be strictly dependent upon the age of

  6. Patterns of puffing activity in the salivary gland chromosomes of Drosophila

    Microsoft Academic Search

    Michael Ashburner

    1972-01-01

    A technique for the short term organ culture of larval salivary glands of D. melanogaster is described. Cultured Puff Stage 1 glands respond to 20-OH ecdysone by initiating the cycle of puffing activity characteristic of late larval development and puparium formation. This puffing cycle involves the sequential activation of at least 125 puffs. Their response to ecdysone allows these puffs

  7. Structure activity relationships of flavonoids as potent alpha-amylase inhibitors.

    PubMed

    Yuan, Erdong; Liu, Benguo; Wei, Qingyi; Yang, Jiguo; Chen, Lei; Li, Qiong

    2014-08-01

    The effects of three flavonoids (quercetin, luteolin, diosmetin) on alpha-amylase were examined by enzymatic kinetics and fluorescence spectroscopy. The three test flavonoids were non-competitive inhibitors of the enzyme. Addition of flavonoids led to fluorescence quenching of alpha-amylase. The quenching was initiated from the formation of a complex between the flavonoids and the enzyme, corresponding to a static quenching process. An alpha-amylase molecule provides a binding site for the test flavonoid. The main binding force was hydrophobic. The decreasing order of inhibition of alpha-amylase by flavonoids and the binding force was luteolin, diosmetin, and quercetin. It is demonstrated that hydroxylation in ring C and methylation of the hydroxyl group in ring B of flavonoids may weaken the binding affinities to alpha-amylase. PMID:25233601

  8. A Study oftheMaizef.Amylase System

    Microsoft Academic Search

    Christiane Lauriere; Christine Doyen; Claudine Thevenot

    1992-01-01

    B-Amylase ofmaize(Zea maysL.) caryopses wasstudied during development andgermination bymeansofenzymic, electropho- retic, andimmunochemical techniques. ,B-Amylase activity in- creased during caryopsis development toamaximumvalue atthe beginning ofthewatercontent plateau (atthis stage theenzyme waslocated primarily within thepericarp) andthendecreased. Almost no,-amylase (activity orantigen) wasfound ineither free orboundforms inthematuremaizecaryopsis. Theactivity in- creased again during seedling growth andreached muchhigher values. Boththealeurone layer (toamajor extent) andthescutel- lumproduced andsecreted 6-amylase

  9. Anticoagulation activity of salivary gland extract of oriental blackfly Simulium indicum

    PubMed Central

    Borah, Subhalaxmi; Naglot, Ashok; Goswami, Sewali; Rahman, Imtiaz; Deka, Manab

    2014-01-01

    Objective To study the morphology of the salivary gland of the female blackfly of the species Simulium indicum (S. indicum) along with protein profile and anticoagulant activity of the salivary gland extract. Methods Sodium dodecyl sulphate polyacrylamide gel electrophoresis was used to analyze the protein profile of the salivary gland extract (SGE) and anticoagulant activities against thrombin, and the extrinsic and intrinsic coagulation pathways were found in S. indicum SGE in the TT, PT and APTT assays, respectively. Results Results revealed that each gland consisted of a cylindrical U-shaped secretory lobe and a more or less spherical reservoir. The protein contents of whole salivary glands were also quantified and the amount of salivary gland proteins in the adult female S. indicum was found out to be approximately 1.12±0.13 µg/female. At least 16 major and several minor protein bands were detected in the female salivary glands. The molecular masses of these major protein bands were estimated at 69, 65, 61, 58, 44, 42, 39, 33, 30, 28, 27, 26, 23, 21, 18 and 16 kDa, consecutively. Anticoagulant activities were found in S. indicum SGE in all the assays. It was found that SGE prolonged human plasma clotting time in a dose-dependent manner. Factor Xa inhibition was shown by the SGE of S. indicum. Percent inhibition value was 93.8. A positive correlation (r=0.89) was observed between total protein and percent inhibition of factor Xa. Conclusions The present study demonstrated that the mode of action of the anticoagulant(s) is mainly on the inhibition of thrombin and factor Xa along with other target factors of the coagulation cascade. PMID:25183091

  10. Antioxidative activity and inhibition of key enzymes linked to type-2 diabetes (?-glucosidase and ?-amylase) by Khaya senegalensis.

    PubMed

    Ibrahim, Mohammed Auwal; Koorbanally, Neil Anthony; Islam, Md Shahidul

    2014-09-01

    This study evaluated the in vitro antioxidative activity of Khaya senegalensis extracts and inhibitory effects of some solvent fractions on ?-glucosidase and ?-amylase activities. The stem bark, root and leaf samples of the plant were sequentially extracted with ethyl acetate, ethanol and water and then tested for antioxidative activity. Our findings revealed that the ethanolic extract of the root had the highest antioxidative activity. Solvent-solvent fractionation of the root ethanolic extract yielded a butanol fraction that showed higher antioxidative activity than other fractions. Furthermore, the butanol fraction had significantly higher (p < 0.05) ?-glucosidase and ?-amylase inhibitory activities with IC50 values of 2.89 ± 0.46 and 97.51 ± 5.72 ?g mL?¹, respectively. Enzyme kinetic studies indicated that the butanol fraction is a non-competitive inhibitor for ?-glucosidase with an inhibition binding constant K(i) of 1.30 ?g mL?¹ and a competitive inhibitor of ?-amylase with a K(i) of 7.50 ?g mL?¹. GC-MS analysis revealed that the butanol fraction contained two chromones, p-anilinophenol and 3-ethyl-5-(3-ethyl-(3H)-benzothiazol-2-ylidene)-2-(p-tolylvinylamino)-4-thiazolidinone. Data obtained in the study suggest that the butanol fraction derived from the ethanolic extract of K. senegalensis root possessed excellent antioxidative as well as ?-glucosidase and a-amylase inhibitory activities while chromones and/or p-anilinophenol could be the main bioactive compounds responsible for the observed activities. PMID:25296677

  11. P2Y2 nucleotide receptor activation enhances the aggregation and self-organization of dispersed salivary epithelial cells.

    PubMed

    El-Sayed, Farid G; Camden, Jean M; Woods, Lucas T; Khalafalla, Mahmoud G; Petris, Michael J; Erb, Laurie; Weisman, Gary A

    2014-07-01

    Hyposalivation resulting from salivary gland dysfunction leads to poor oral health and greatly reduces the quality of life of patients. Current treatments for hyposalivation are limited. However, regenerative medicine to replace dysfunctional salivary glands represents a revolutionary approach. The ability of dispersed salivary epithelial cells or salivary gland-derived progenitor cells to self-organize into acinar-like spheres or branching structures that mimic the native tissue holds promise for cell-based reconstitution of a functional salivary gland. However, the mechanisms involved in salivary epithelial cell aggregation and tissue reconstitution are not fully understood. This study investigated the role of the P2Y2 nucleotide receptor (P2Y2R), a G protein-coupled receptor that is upregulated following salivary gland damage and disease, in salivary gland reconstitution. In vitro results with the rat parotid acinar Par-C10 cell line indicate that P2Y2R activation with the selective agonist UTP enhances the self-organization of dispersed salivary epithelial cells into acinar-like spheres. Other results indicate that the P2Y2R-mediated response is dependent on epidermal growth factor receptor activation via the metalloproteases ADAM10/ADAM17 or the ?5?1 integrin/Cdc42 signaling pathway, which leads to activation of the MAPKs JNK and ERK1/2. Ex vivo data using primary submandibular gland cells from wild-type and P2Y2R(-/-) mice confirmed that UTP-induced migratory responses required for acinar cell self-organization are mediated by the P2Y2R. Overall, this study suggests that the P2Y2R is a promising target for salivary gland reconstitution and identifies the involvement of two novel components of the P2Y2R signaling cascade in salivary epithelial cells, the ?5?1 integrin and the Rho GTPase Cdc42. PMID:24760984

  12. In vitro antioxidant and pancreatic ?-amylase inhibitory activity of isolated fractions from water extract of Qingzhuan tea.

    PubMed

    Cheng, Qian; Cai, Shengbao; Ni, Dejiang; Wang, Ruojun; Zhou, Feng; Ji, Baoping; Chen, Yuqiong

    2015-02-01

    In the present work, Qingzhuan tea, a unique dark tea produced by post-fermentation technology, was selected to investigate its antioxidant and pancreatic ?-amylase inhibiting activities. Water extract of Qingzhuan tea was successively isolated by solvent partitioning procedures to obtain chloroform, ethyl acetate, n-butanol, sediment and residual aqua fractions. Of different fractions, the ethyl acetate fraction (QEF) had the highest total polyphenols and catechins contents, demonstrated the strongest DPPH radical scavenging activity and exhibited the greatest inhibitory effect on porcine pancreatic ?-amylase activity in vitro. Further separation of QEF by a Sephadex LH-20 column generated eight subfractions (QEF1-QEF8), with QEF8 being the most active subfraction based on the assays above mentioned. The major active components in QEF8 were identified as catechins EGCG and ECG by LC-MS analysis, with contents of 22.29 % and 11.11 % respectively. Inhibitory effects of catechin standards EGCG and ECG on porcine pancreatic ?-amylase activity were also observed. In conclusion, Qingzhuan tea or its water extract could be potentially used as complementary therapy ingredients for diabetes treatment through lowering postprandial blood glucose, and catechins EGCG and ECG may be the most efficient components in the water extract. PMID:25694702

  13. Detergent-compatible bacterial amylases.

    PubMed

    Niyonzima, Francois N; More, Sunil S

    2014-10-01

    Proteases, lipases, amylases, and cellulases are enzymes used in detergent formulation to improve the detergency. The amylases are specifically supplemented to the detergent to digest starchy stains. Most of the solid and liquid detergents that are currently manufactured contain alkaline enzymes. The advantages of using alkaline enzymes in the detergent formulation are that they aid in removing tough stains and the process is environmentally friendly since they reduce the use of toxic detergent ingredients. Amylases active at low temperature are preferred as the energy consumption gets reduced, and the whole process becomes cost-effective. Most microbial alkaline amylases are used as detergent ingredients. Various reviews report on the production, purification, characterization, and application of amylases in different industry sectors, but there is no specific review on bacterial or fungal alkaline amylases or detergent-compatible amylases. In this mini-review, an overview on the production and property studies of the detergent bacterial amylases is given, and the stability and compatibility of the alkaline bacterial amylases in the presence of the detergents and the detergent components are highlighted. PMID:25129040

  14. Purification, biochemical characterisation and partial primary structure of a new alpha-amylase inhibitor from Secale cereale (rye).

    PubMed

    Iulek, J; Franco, O L; Silva, M; Slivinski, C T; Bloch, C; Rigden, D J; Grossi de Sá, M F

    2000-01-01

    Plant alpha-amylase inhibitors show great potential as tools to engineer resistance of crop plants against pests. Their possible use is, however, complicated by the observed variations in specificity of enzyme inhibition, even within closely related families of inhibitors. Better understanding of this specificity depends on modelling studies based on ample structural and biochemical information. A new member of the alpha-amylase inhibitor family of cereal endosperm has been purified from rye using two ionic exchange chromatography steps. It has been characterised by mass spectrometry, inhibition assays and N-terminal protein sequencing. The results show that the inhibitor has a monomer molecular mass of 13,756 Da, is capable of dimerisation and is probably glycosylated. The inhibitor has high homology with the bifunctional alpha-amylase/trypsin inhibitors from barley and wheat, but much poorer homology with other known inhibitors from rye. Despite the homology with bifunctional inhibitors, this inhibitor does not show activity against mammalian or insect trypsin, although activity against porcine pancreatic, human salivary, Acanthoscelides obtectus and Zabrotes subfasciatus alpha-amylases was observed. The inhibitor is more effective against insect alpha-amylases than against mammalian enzymes. It is concluded that rye contains a homologue of the bifunctional alpha-amylase/trypsin inhibitor family without activity against trypsins. The necessity of exercising caution in assigning function based on sequence comparison is emphasised. PMID:11137459

  15. Rubusuaviins A-F, monomeric and oligomeric ellagitannins from Chinese sweet tea and their alpha-amylase inhibitory activity.

    PubMed

    Li, Haizhou; Tanaka, Takashi; Zhang, Ying-Jun; Yang, Chong-Ren; Kouno, Isao

    2007-09-01

    Six new ellagitannins herein, rubusuaviins A-F, were isolated from the aqueous acetone extract of Chinese sweet tea (Tien-cha, dried leaves of Rubus suavissimus S. LEE) together with seven known tannins. Rubusuaviin A was characterized as 1-O-galloyl-2,3-O-(S)-HHDP-4,6-O-(S)-sanguisorboyl-beta-D-glucopyranose. Rubusuaviins B, C, and E are dimeric, trimeric, and tetrameric ellagitannins, respectively, in which the sanguisorboyl groups were connected ellagitannin units. Rubusuaviins D and F were desgalloyl derivatives of rubusuaviins C and E, respectively. The inhibition of alpha-amylase activity by rubusuaviins and related ellagitannins was compared. Ellagitannins with beta-galloyl groups at the glucose C-1 positions showed stronger inhibition compared with the alpha-galloyl and desgalloyl compounds. The molecular weight of these compounds was not important for the inhibition of alpha-amylase activity. PMID:17827756

  16. The interplay of ?-amylase and amyloglucosidase activities on the digestion of starch in in vitro enzymic systems.

    PubMed

    Warren, Frederick J; Zhang, Bin; Waltzer, Gina; Gidley, Michael J; Dhital, Sushil

    2015-03-01

    In vitro hydrolysis assays are a key tool in understanding differences in rate and extent of digestion of starchy foods. They offer a greater degree of simplicity and flexibility than dynamic in vitro models or in vivo experiments for quantifiable, mechanistic exploration of starch digestion. In the present work the influence of ?-amylase and amyloglucosidase activities on the digestion of maize and potato starch granules was measured using both glucose and reducing sugar assays. Data were analysed through initial rates of digestion, and by 1st order kinetics, utilising logarithm of slope (LOS) plots. The rate and extent of starch digestion was dependent on the activities of both enzymes and the type of starch used. Potato required more enzyme than maize to achieve logarithmic reaction curves, and complete digestion. The results allow targeted design of starch digestion experiments through a thorough understanding of the contributions of ?-amylase and amyloglucosidase to digestion rates. PMID:25498625

  17. Pharmacological Activation of the EDA/EDAR Signaling Pathway Restores Salivary Gland Function following Radiation-Induced Damage

    PubMed Central

    Hill, Grace; Headon, Denis; Harris, Zoey I.; Huttner, Kenneth; Limesand, Kirsten H.

    2014-01-01

    Radiotherapy of head and neck cancers often results in collateral damage to adjacent salivary glands associated with clinically significant hyposalivation and xerostomia. Due to the reduced capacity of salivary glands to regenerate, hyposalivation is treated by substitution with artificial saliva, rather than through functional restoration of the glands. During embryogenesis, the ectodysplasin/ectodysplasin receptor (EDA/EDAR) signaling pathway is a critical element in the development and growth of salivary glands. We have assessed the effects of pharmacological activation of this pathway in a mouse model of radiation-induced salivary gland dysfunction. We report that post-irradiation administration of an EDAR-agonist monoclonal antibody (mAbEDAR1) normalizes function of radiation damaged adult salivary glands as determined by stimulated salivary flow rates. In addition, salivary gland structure and homeostasis is restored to pre-irradiation levels. These results suggest that transient activation of pathways involved in salivary gland development could facilitate regeneration and restoration of function following damage. PMID:25409170

  18. Hypoglycemic activity of Pyrus biossieriana Buhse leaf extract and arbutin: Inhibitory effects on alpha amylase and alpha glucosidase

    PubMed Central

    Yousefi, Fatemeh; Mahjoub, Soleiman; Pouramir, Mahdi; Khadir, Fatemeh

    2013-01-01

    Background: The mechanism of hypoglycemic and hypolipidemic activities of Pyrus biossieriana Buhse leaf extract (PbBLE) and its phytochemical component arbutin, have not been well determined. The present study was performed to understand the hypoglycemic activity mechanisms of pbBLE and arbutin more clearly. Methods: In vitro enzymatic carbohydrate digestion with PbBLE and arbutin was assessed using ?-amylase and ?-glucosidase powders. The enzyme solutions were premixed with PbBLE and arbutin at different concentrations (0.1, 1, 10 and 100 mg/ml). Substrate solutions and colorimetric reagents were added to the reaction. The release of glucose was determined by spectrophotometric method. Acarbose was used as the positive control. Results: The extract (10, 100 mg/ ml) completely inhibit ?- amylase and ?- glucosidase activities. The extract produced higher reduction of ?-amylase and ?-glucosidase activity than arbutin. Inhibition at various concentrations (0.1, 1, 10, 100 mg/ml) were significantly different (p<0.05). Conclusion: Our results exhibited that both the extract and arbutin were able to suppress the enzymes strongly. PMID:24294470

  19. The ingredients in Saengshik, a formulated health food, inhibited the activity of ?-amylase and ?-glucosidase as anti-diabetic function

    PubMed Central

    Kim, Misook; Kim, Eunji; Kwak, Han Sub

    2014-01-01

    BACKGROUND/OBJECTIVES We investigated total 26 ingredients of Saengshik which will be commercially produced as an anti-diabetic dietary supplement. SUBJECTS/METHODS Thirteen vegetables, nine cereals, three legumes and one seed were extracted with aqueous ethanol for 2 h at 60?, and evaluated for their inhibitory effects against ?-amylase and ?-glucosidase and for total phenolic and flavonoid contents. RESULTS All ingredients inhibited ?-amylase activity except cabbage. Strong inhibitory activity of ?-amylase was observed in leek, black rice, angelica and barley compared with acarbose as a positive control. Stronger inhibition of ?-glucosidase activity was found in small water dropwort, radish leaves, sorghum and cabbage than acarbose. All Saengshik ingredients suppressed ?-glucosidase activity in the range of 0.3-60.5%. Most ingredients contained total phenols which were in the range of 1.2-229.4 mg gallic acid equivalent/g dried extract. But, total phenolic contents were not observed in carrot, pumpkin and radish. All ingredients contained flavonoid in the range of 11.6-380.7 mg catechin equivalent/g dried extract. CONCLUSIONS Our results demonstrate that Saengshik containing these ingredients would be an effective dietary supplement for diabetes. PMID:25324943

  20. Antiviral activity of salivary microRNAs for ophthalmic herpes zoster.

    PubMed

    Irmak, M Kemal; Erdem, Uzeyir; Kubar, Ayhan

    2012-01-01

    Ophthalmic herpes zoster is a common ocular infection caused by the varicella-zoster virus (VZV). Viral mRNA transcripts play a major role in the replicative cycle of the virus and current antiviral agents have little effect in preventing and treating the complications. Therapeutic use of saliva for certain painful ocular diseases such as ophthalmic herpes zoster is a well-known public practice in our region. We thought that antiviral activity of saliva may stem from salivary microvesicles and we aimed to look for molecules with antiviral activity in these vesicles. As a possible candidate for antiviral activity, salivary microvesicles contain at least 20 microRNAs (miRNAs), small noncoding RNAs, which suppress the translation of target mRNAs. miRNAs not only participate in maintenance of normal cell functions, but are also involved in host-virus interactions and limit the replication of certain virus types. Thus, miRNA gene therapy by targeting mRNAs required for VZV survival may find a niche in the treatment of ophthalmic herpes zoster. But, how could salivary microvesicles reach into the corneal cells to demonstrate their antiviral activity. We suggest that human salivary microvesicles can be effective carriers of miRNA for corneal cells, because they contain a molecular machinery for vesicle trafficking and fusion allowing them to be endocytosed by target cells. After binding to the plasma membrane, microvesicles seem to enter into the corneal cells through the clathrin-mediated endocytosis. In the cytosol, human salivary miRNAs base-pair with specific viral mRNAs and inhibit their translation, thus limiting the replication of the virus. PMID:22676898

  1. Enzymatic activities in different strains isolated from healthy and brittle leaf disease affected date palm leaves: study of amylase production conditions.

    PubMed

    Mouna, Jrad; Imen, Fendri; Choba Ines, Ben; Nourredine, Drira; Adel, Kadri; Néji, Gharsallah

    2015-02-01

    The present study aimed to investigate and compare the enzymatic production of endophytic bacteria isolated from healthy and brittle leaf disease affected date palm leaves (pectinase, cellulase, lipase, and amylase). The findings revealed that the enzymatic products from the bacterial isolates of healthy date palm leaves were primarily 33% amylolytic enzyme, 33 % cellulase, 25 % pectinase, and 25 % lipase. The isolates from brittle leaf disease date palm leaves, on the other hand, were noted to produce 16 % amylolytic enzyme, 20 % cellulose, 50 % pectinase, and 50 % lipase. The effects of temperature and pH on amylase, pectinase, and cellulose activities were investigated. The Bacillus subtilis JN934392 strain isolated from healthy date palm leaves produced higher levels of amylase activity at pH 7. A Box Behnken Design (BBD) was employed to optimize amylase extraction. Maximal activity was observed at pH and temperature ranges of pH 6-6.5 and 37-39 °C, respectively. Under those conditions, amylase activity was noted to be attained 9.37 U/ml. The results showed that the enzyme was able to maintain more than 50 % of its activity over a temperature range of 50-80 °C, with an optimum at 70 °C. This bacterial amylase showed high activity compared to other bacteria, which provides support for its promising candidacy for future industrial application. PMID:25432343

  2. Searching for Amylase

    NSDL National Science Digital Library

    Keith D. Stanley (Beloit College; Biology)

    2006-05-20

    Information about sequence, structure, and active sites for many proteins is accessible online. This real world application of bioinformatics introduces us to amylases and asks us to explore functionally similar enzymes from markedly divergent organisms. Here, we search for microbial enzymes useful in the starch processing industry. Several tools useful for dealing with sequence information are introduced through the Biology Workbench.Information about sequence, structure, and active sites for many proteins is accessible online. This real world application of bioinformatics introduces us to amylases and asks us to explore functionally similar enzymes from markedly divergent organisms. Here, we search for microbial enzymes useful in the starch processing industry. Several tools useful for dealing with sequence information are introduced through the Biology Workbench. * investigate the use of bioinformatics to identify potentially useful microbial amylases for industry

  3. Inhibition of key enzymes linked to obesity by preparations from Mediterranean dietary plants: effects on ?-amylase and pancreatic lipase activities.

    PubMed

    2013-12-01

    One of the most important strategy in the treatment of obesity includes the development of nutrient digestion and absorption inhibitors. Inhibition of digestive enzymes is one of the most widely studied mechanisms used to determine the potential efficacy of natural products as hypolipidemic and hypoglycaemic agents. In vitro studies here reported were performed to evaluate the inhibitory activity of five species(as hydroalcoholic extracts) of edible plants from Calabria region (Italy) on amylase and lipase by monitoring the hydrolysis of p-NPC and the hydrolysis of glycoside bonds indigestible carbohydrate foods. The formulation obtained from Clematis vitalba L. exhibited the strongest inhibitory effect on pancreatic lipase (IC50=0.99 mg/ml) and on ?-amylase(IC50=31.52 ?g/ml). In order to explore metabolome production HPTLC analysis of the extracts was performed, revealing the predominance of (±)-catechin, caffeic acid and chlorogenic acid in C. vital ba formulation at concentration of 23.18±3.14,13.63±0.65 and 18.88±0.76 mg/g, respectively. GC/MS analysis was used to identify fatty acids and terpene composition. PMID:24122547

  4. Componential Profile and Amylase Inhibiting Activity of Phenolic Compounds from Calendula officinalis L. Leaves

    PubMed Central

    Olennikov, Daniil N.; Kashchenko, Nina I.

    2014-01-01

    An ethanolic extract and its ethyl acetate-soluble fraction from leaves of Calendula officinalis L. (Asteraceae) were found to show an inhibitory effect on amylase. From the crude extract fractions, one new phenolic acid glucoside, 6?-O-vanilloyl-?-D-glucopyranose, was isolated, together with twenty-four known compounds including five phenolic acid glucosides, five phenylpropanoids, five coumarins, and nine flavonoids. Their structures were elucidated based on chemical and spectral data. The main components, isoquercitrin, isorhamnetin-3-O-?-D-glucopyranoside, 3,5-di-O-caffeoylquinic acid, and quercetin-3-O-(6??-acetyl)-?-D-glucopyranoside, exhibited potent inhibitory effects on amylase. PMID:24683352

  5. Association of Novel Domain in Active Site of Archaic Hyperthermophilic Maltogenic Amylase from Staphylothermus marinus*

    PubMed Central

    Jung, Tae-Yang; Li, Dan; Park, Jong-Tae; Yoon, Se-Mi; Tran, Phuong Lan; Oh, Byung-Ha; Jane?ek, Štefan; Park, Sung Goo; Woo, Eui-Jeon; Park, Kwan-Hwa

    2012-01-01

    Staphylothermus marinus maltogenic amylase (SMMA) is a novel extreme thermophile maltogenic amylase with an optimal temperature of 100 °C, which hydrolyzes ?-(1–4)-glycosyl linkages in cyclodextrins and in linear malto-oligosaccharides. This enzyme has a long N-terminal extension that is conserved among archaic hyperthermophilic amylases but is not found in other hydrolyzing enzymes from the glycoside hydrolase 13 family. The SMMA crystal structure revealed that the N-terminal extension forms an N? domain that is similar to carbohydrate-binding module 48, with the strand-loop-strand region forming a part of the substrate binding pocket with several aromatic residues, including Phe-95, Phe-96, and Tyr-99. A structural comparison with conventional cyclodextrin-hydrolyzing enzymes revealed a striking resemblance between the SMMA N? domain position and the dimeric N domain position in bacterial enzymes. This result suggests that extremophilic archaea that live at high temperatures may have adopted a novel domain arrangement that combines all of the substrate binding components within a monomeric subunit. The SMMA structure provides a molecular basis for the functional properties that are unique to hyperthermophile maltogenic amylases from archaea and that distinguish SMMA from moderate thermophilic or mesophilic bacterial enzymes. PMID:22223643

  6. In vitro ?-amylase inhibitory activity and in vivo hypoglycemic effect of methanol extract of Citrus macroptera Montr. fruit

    PubMed Central

    Uddin, Nizam; Hasan, Md. Rakib; Hossain, Md. Monir; Sarker, Arjyabrata; Hasan, A.H.M. Nazmul; Islam, A.F.M. Mahmudul; Chowdhury, Mohd. Motaher H.; Rana, Md. Sohel

    2014-01-01

    Objective To investigate the therapeutic effects of methanol extract of Citrus macroptera Montr.fruit in ?-amylase inhibitory activity (in vitro) and hypoglycemic activity in normal and glucose induced hyperglycemic rats (in vivo). Methods Fruits of Citrus macroptera without rind was extracted with pure methanol following cold extraction and tested for presence of phytochemical constituents, ?-amylase inhibitory activity, and hypoglycemic effect in normal rats and glucose induced hyperglycemic rats. Results Presence of saponin, steroid and terpenoid were identified in the extract. The results showed that fruit extract had moderate ?-amylase inhibitory activity [IC50 value=(3.638±0.190) mg/mL] as compared to acarbose. Moreover at 500 mg/kg and 1?000 mg/kg doses fruit extract significantly (P<0.05 and P<0.01 respectively) reduced fasting blood glucose level in normal rats as compared to glibenclamide (5 mg/kg). In oral glucose tolerance test, 500 mg/kg dose significantly reduced blood glucose level (P<0.05) at 2 h but 1?000 mg/kg dose significantly reduced blood glucose level at 2 h and 3 h (P<0.05 and P<0.01 respectively) whereas glibenclamide (5 mg/kg) significantly reduced glucose level at every hour after administration. Overall time effect is also considered extremely significant with F value=23.83 and P value=0.0001 in oral glucose tolerance test. Conclusion These findings suggest that the plant may be a potential source for the development of new oral hypoglycemic agent. PMID:25182949

  7. Enzymatic activity and immunoreactivity of Aca s 4, an alpha-amylase allergen from the storage mite Acarus siro

    PubMed Central

    2012-01-01

    Background Enzymatic allergens of storage mites that contaminate stored food products are poorly characterized. We describe biochemical and immunological properties of the native alpha-amylase allergen Aca s 4 from Acarus siro, a medically important storage mite. Results A. siro produced a high level of alpha-amylase activity attributed to Aca s 4. This enzyme was purified and identified by protein sequencing and LC-MS/MS analysis. Aca s 4 showed a distinct inhibition pattern and an unusual alpha-amylolytic activity with low sensitivity to activation by chloride ions. Homology modeling of Aca s 4 revealed a structural change in the chloride-binding site that may account for this activation pattern. Aca s 4 was recognized by IgE from house dust mite-sensitive patients, and potential epitopes for cross-reactivity with house dust mite group 4 allergens were found. Conclusions We present the first protein-level characterization of a group 4 allergen from storage mites. Due to its high production and IgE reactivity, Aca s 4 is potentially relevant to allergic hypersensitivity. PMID:22292590

  8. The developmental course of salivary alpha-amylase and cortisol from 12 to 36 months: Relations with early poverty and later behavior problems.

    PubMed

    Hill-Soderlund, Ashley L; Holochwost, Steven J; Willoughby, Michael T; Granger, Douglas A; Gariépy, Jean-Louis; Mills-Koonce, W Roger; Cox, Martha J

    2015-02-01

    This study examined the development of baseline autonomic nervous system (ANS) and hypothalamic-pituitary-adrenal (HPA) physiological activity from 12 to 36 months as well as antecedents (poverty) and consequents (behavior problems) of individual differences in physiological development. Children (N=179; 50% poor; 56% African American; 52% male) provided saliva samples at 12, 18, 24, 30, and 36 months of age. Latent growth curve models indicated that nonlinear change was evident for both sAA and cortisol, with sAA increasing and cortisol decreasing with age. Children residing in poor households exhibited lower initial levels of sAA, but not cortisol. African-American children showed slightly smaller decreases in cortisol over time. Initial levels of sAA predicted higher levels of internalizing behaviors at 36 months and both initial levels of and total change in sAA predicted higher levels of externalizing behaviors at 36 months. There was no evidence that sAA or cortisol mediated the relationship between poverty and later behavior problems. PMID:25245323

  9. Effect of codon-optimized E. coli signal peptides on recombinant Bacillus stearothermophilus maltogenic amylase periplasmic localization, yield and activity.

    PubMed

    Samant, Shalaka; Gupta, Gunja; Karthikeyan, Subbulakshmi; Haq, Saiful F; Nair, Ayyappan; Sambasivam, Ganesh; Sukumaran, Sunilkumar

    2014-09-01

    Recombinant proteins can be targeted to the Escherichia coli periplasm by fusing them to signal peptides. The popular pET vectors facilitate fusion of target proteins to the PelB signal. A systematic comparison of the PelB signal with native E. coli signal peptides for recombinant protein expression and periplasmic localization is not reported. We chose the Bacillus stearothermophilus maltogenic amylase (MA), an industrial enzyme widely used in the baking and brewing industry, as a model protein and analyzed the competence of seven, codon-optimized, E. coli signal sequences to translocate MA to the E. coli periplasm compared to PelB. MA fusions to three of the signals facilitated enhanced periplasmic localization of MA compared to the PelB fusion. Interestingly, these three fusions showed greatly improved MA yields and between 18- and 50-fold improved amylase activities compared to the PelB fusion. Previously, non-optimal codon usage in native E. coli signal peptide sequences has been reported to be important for protein stability and activity. Our results suggest that E. coli signal peptides with optimal codon usage could also be beneficial for heterologous protein secretion to the periplasm. Moreover, such fusions could even enhance activity rather than diminish it. This effect, to our knowledge has not been previously documented. In addition, the seven vector platform reported here could also be used as a screen to identify the best signal peptide partner for other recombinant targets of interest. PMID:25038884

  10. Study of phenolic content and urease and alpha-amylase inhibitory activities of methanolic extract of Rumex acetosella roots and its sub-fractions in different solvents.

    PubMed

    Ahmed, Dildar; Mughal, Qaria Mumtaz; Younas, Saba; Ikram, Muhammad

    2013-05-01

    The present study aimed to establish relationship between urease and alpha-amylase inhibitory activities on the one hand and on the other between anti-enzymatic activities and total phenolic contents of the methanolic extract of roots of Rumex acetosella and its fractions in various solvents. The methanolic extract and its fractions in chloroform, ethyl acetate, n-butanol and water showed remarkable inhibitory activities against both urease and alpha-amylase, there was a close correspondence between urease and alpha-amylase inhibitory activities of the plant samples. The n-butanol fraction which had the highest total phenolic content (252.19 ± 2.32 µg of Gallic Acid Equivalents/mg of dry mass of the sample) showed prominent activity against both urease and alpha-amylase indicating a possible role of phenolics in inhibiting the activities of these enzymes. The samples displayed enzyme inhibitory activities in a dose dependent manner and their effectiveness was comparable with that of the standards, thiourea (for urease) and acarbose (for alpha-amylase). The samples were manifold more effective against urease than alpha-amylase; 2.8 mg/mL of MeOH extract produced about 81% inhibition in alpha-amylase activity, while only 10 µg/mL of the extract was required to create the same inhibition in urease activity. The IC50 values of methanolic, chloroform, ethyl acetate, n-butanolic, aqueous and standard solutions were 1.29, 1.31, 1.90, 1.38, 0.85 and 1.20 (mg/mL) respectively against alpha-amylase and 0.99, 3.89, 1.76, 0.91, 0.85 and 0.97 (?g/mL) respectively against urease. The total phenolic content in MeOH, hexane, chloroform, ethyl acetate, n-butanol and water fractions was 108.88 ± 2.65, 43.70 ± 1.90, 34.44 ± 2.30, 230.71 ± 1.78, 252.19 ± 2.32 and 94.07 ± 2.25 respectively. PMID:23625429

  11. Salivary cholinesterase activity in children with organic and convential diets

    EPA Science Inventory

    Objective: Previous efforts to determine the health effects of pesticides have focused on quantifying acetylcholinesterase activity in blood. However, since blood draws can be difficult in young children, saliva biomonitoring has recently been explored as a feasible alternative....

  12. The relationship between objectively measured physical activity, salivary cortisol, and the metabolic syndrome score in girls.

    PubMed

    DuBose, Katrina D; McKune, Andrew J

    2014-08-01

    The relationship between physical activity levels, salivary cortisol, and the metabolic syndrome (MetSyn) score was examined. Twenty-three girls (8.4 ± 0.9 years) had a fasting blood draw, waist circumference and blood pressure measured, and wore an ActiGraph accelerometer for 5 days. Saliva samples were collected to measure cortisol levels. Previously established cut points estimated the minutes spent in moderate, vigorous, and moderate-to-vigorous physical activity. A continuous MetSyn score was created from blood pressure, waist circumference, high-density-lipoprotein (HDL), triglyceride, and glucose values. Correlation analyses examined associations between physical activity, cortisol, the MetSyn score, and its related components. Regression analysis examined the relationship between cortisol, the MetSyn score, and its related components adjusting for physical activity, percent body fat, and sexual maturity. Vigorous physical activity was positively related with 30 min post waking cortisol values. The MetSyn score was not related with cortisol values after controlling for confounders. In contrast, HDL was negatively related with 30 min post waking cortisol. Triglyceride was positively related with 30 min post waking cortisol and area under the curve. The MetSyn score and many of its components were not related to cortisol salivary levels even after adjusting for physical activity, body fat percentage, and sexual maturity. PMID:24722755

  13. Gamma irradiation of sorghum flour: Effects on microbial inactivation, amylase activity, fermentability, viscosity and starch granule structure

    NASA Astrophysics Data System (ADS)

    Mukisa, Ivan M.; Muyanja, Charles M. B. K.; Byaruhanga, Yusuf B.; Schüller, Reidar B.; Langsrud, Thor; Narvhus, Judith A.

    2012-03-01

    Malted and un-malted sorghum ( Sorghum bicolor (L.) Moench) flour was gamma irradiated with a dose of 10 kGy and then re-irradiated with 25 kGy. The effects of irradiation on microbial decontamination, amylase activity, fermentability (using an amylolytic L. plantarum MNC 21 strain), starch granule structure and viscosity were determined. Standard methods were used during determinations. The 10 kGy dose had no effect on microbial load of un-malted flour but reduced that of malted flour by 3 log cycles. Re-irradiation resulted in complete decontamination. Irradiation of malt caused a significant ( p<0.05) reduction in alpha and beta amylase activity (22% and 32%, respectively). Irradiation of un-malted flour increased the rates of utilization of glucose and maltose by 53% and 100%, respectively, during fermentation. However, microbial growth, rate of lactic acid production, final lactic acid concentration and pH were not affected. Starch granules appeared normal externally even after re-irradiation, however, granules ruptured and dissolved easily after hydration and gelatinization. Production of high dry matter density porridge (200 g dry matter/L) with a viscosity of 3500 cP was achieved by irradiation of un-malted flout at 10 kGy. Gamma irradiation can be used to decontaminate flours and could be utilized to produce weaning porridge from sorghum.

  14. NMR assignment of the amylase-binding protein A from Streptococcus parasanguinis.

    PubMed

    Liu, Bing; Zhu, Fan; Wu, Hui; Matthews, Stephen

    2015-04-01

    Streptococcus parasanguinis is a primary colonizer of tooth surfaces in the oral cavity. Amylase-binding protein A (AbpA) from S. parasanguinis is responsible for the recruitment of salivary amylase to bacterial surface, which plays an important role in the development of oral biofilms. Here, we describe the essentially complete NMR assignments for AbpA. PMID:25016927

  15. 21 CFR 862.1070 - Amylase test system.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...measure the activity of the enzyme amylase in serum and urine. Amylase measurements are used primarily for the diagnosis and treatment of pancreatitis (inflammation of the pancreas). (b) Classification. Class...

  16. Comparisons of amylolytic enzyme activities and ß-amylases with differing Bmy1 intron III alleles to osmolyte concentration and malt extract during congress mashing with North American barley cultivars

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was conducted to determine the relationships between patterns of activity of malt amylolytic enzymes (a-amylase, ß-amylase, and limit dextrinase) and the patterns of osmolyte concentration (OC) and malt extract (ME) production in two- and six-row North American barley cultivars over the c...

  17. Thermal adaptation of ?-amylases: a review.

    PubMed

    Hiteshi, Kalpana; Gupta, Reena

    2014-11-01

    The temperature adaptation of ?-amylase can be gained by different adjustments in protein structure with consecutive effects on the stability and flexibility of the protein. In this review, meso, thermo and cold-active ?-amylases have been compared with respect to their structure and intramolecular interactions. With decrease in temperature, the number of ionic interactions also decreases, leading to greater flexibility of proteins. It has also been observed that the proline and arginine content is higher in thermophilic amylases as compared to meso and psychrophilic amylases, increasing the rigidity and structural stability of protein molecule. PMID:25116053

  18. Age-independent increases in male salivary testosterone during horticultural activity among Tsimane forager-farmers

    PubMed Central

    TRUMBLE, BENJAMIN C; CUMMINGS, DANIEL K; O’CONNOR, KATHLEEN A; HOLMAN, DARRYL J; SMITH, ERIC A; KAPLAN, HILLARD S; GURVEN, MICHAEL D

    2013-01-01

    Testosterone plays an important role in mediating male reproductive trade-offs in many vertebrate species, augmenting muscle and influencing behavior necessary for male-male competition and mating-effort. Among humans, testosterone may also play a key role in facilitating male provisioning of offspring as muscular and neuromuscular performance are deeply influenced by acute changes in testosterone. This study examines acute changes in salivary testosterone among 63 Tsimane men ranging in age from 16–80 (mean 38.2) years during one-hour bouts of tree-chopping while clearing horticultural plots. The Tsimane forager-horticulturalists living in the Bolivian Amazon experience high energy expenditure associated with food production, have high levels of parasites and pathogens, and display significantly lower baseline salivary testosterone than age-matched US males. Mixed-effects models controlling for BMI and time of specimen collection reveal increased salivary testosterone (p<0.001) equivalent to a 48.6% rise, after one hour of tree chopping. Age had no effect on baseline (p=0.656) or change in testosterone (p=0.530); self-reported illness did not modify testosterone change (p=0.488). A comparison of these results to the relative change in testosterone during a competitive soccer tournament in the same population reveals larger relative changes in testosterone following resource production (tree chopping), compared to competition (soccer). These findings highlight the importance of moving beyond a unidimensional focus on changes in testosterone and male-male aggression to investigate the importance of testosterone-behavior interactions across additional male fitness-related activities. Acutely increased testosterone during muscularly intensive horticultural food production may facilitate male productivity and provisioning. PMID:24187482

  19. Curcumin Treatment Suppresses IKK? Kinase Activity of Salivary Cells of Patients with Head and Neck Cancer: A Pilot Study

    PubMed Central

    Kim, Suejung G.; Veena, Mysore S.; Basak, Saroj K.; Han, Eugene; Tajima, Tracey; Gjertson, David W.; Starr, Joshua; Eidelman, Ofer; Pollard, Harvey B.; Srivastava, Meera; Srivatsan, Eri S.; Wang, Marilene B.

    2011-01-01

    Purpose To determine whether curcumin would inhibit IKK? kinase activity and suppress expression of proinflammatory cytokines in head and neck cancer (HNSCC) patients. Experimental Design Saliva was collected before and after subjects chewed curcumin tablets. Protein was extracted and IKK? kinase activity measured. IL-6 and IL-8 levels in the salivary supernatants were measured using ELISA. IL-6, IL-8 and other interleukins were also measured independently with ELISA to confirm the inhibitory effect of curcumin on expression and secretion of salivary cytokines. Results Curcumin treatment led to a reduction in IKK? kinase activity in the salivary cells of HNSCC patients (p<0.05). Treatment of UM-SCC1 cells with curcumin as well as with post curcumin salivary supernatant showed a reduction of IKK? kinase activity. Significant reduction of IL-8 levels (p<0.05) was seen in post curcumin samples from patients with dental caries. Although there was reduced IL-8 expression in 8 of 21 post curcumin samples of HNSCC patients, the data did not reach statistical significance. Saliva samples from HNSCC patients were also analyzed in a blinded fashion for expression of cytokines. IL-10, IFN-?, IL-12p70, and IL-2 clustered together, and GM-CSF and TNF-? clustered together. Log10 ratio analysis demonstrated decrease in expression of all nine cytokines in both the salivary supernatant and salivary cells of curcumin treated samples. Conclusions Curcumin inhibited IKK? kinase activity in the saliva of HNSCC patients and this inhibition correlated with reduced expression of a number of cytokines. IKK? kinase could be a useful biomarker for detecting the effect of curcumin in head and neck cancer. PMID:21821700

  20. Evaluation of the specificity and effectiveness of selected oral hygiene actives in salivary biofilm microcosms.

    PubMed

    Ledder, Ruth G; Sreenivasan, Prem K; DeVizio, William; McBain, Andrew J

    2010-12-01

    The microbiological effects of biocidal products used for the enhancement of oral hygiene relate to the active compound(s) as well as other formulation components. Here, we test the specificities of selected actives in the absence of multiple excipients. Salivary ecosystems were maintained in tissue culture plate-based hydroxyapatite disc models (HDMs) and modified drip-flow biofilm reactors (MDFRs). Test compounds stannous fluoride (SF), SDS, triclosan (TCS), zinc lactate (ZL) and ZL with SF in combination (ZLSF) were delivered to the HDMs once and four times daily for 6 days to MDFRs. Plaques were characterized by differential viable counting and PCR-denaturing gradient gel electrophoresis (DGGE). TCS and SDS were the most effective compounds against HDM plaques, significantly reducing total viable counts (P<0.05), whilst SF, ZL and ZLSF were comparatively ineffective. TCS exhibited specificity for streptococci (P<0.01) and Gram-negative anaerobes (P<0.01) following a single dosing and also on repeated dosing in MDFRs. In contrast to single exposures, multiple dosing with ZLSF also significantly reduced all bacterial groups, whilst SF and ZL caused significant but transient reductions. According to PCR-DGGE analyses, significant (P<0.05) reductions in eubacterial diversity occurred following 6 day dosing with both TCS and ZLSF. Concordance of MDFR eubacterial profiles with salivary inocula ranged between 58 and 97%. TCS and ZL(SF) exhibited similar specificities to those reported for formulations. TCS was the most potent antibacterial, after single and multiple dosage regimens. PMID:20724503

  1. Dietary effects of harmine, a ?-carboline alkaloid, on development, energy reserves and ?-amylase activity of Plodia interpunctella Hübner (Lepidoptera: Pyralidae)

    PubMed Central

    Bouayad, Noureddin; Rharrabe, Kacem; Lamhamdi, Mostafa; Nourouti, Naima Ghailani; Sayah, Fouad

    2010-01-01

    The physiological and developmental effects of harmine, a ?-carboline alkaloid, on the insect pest Plodia interpunctella have been analyzed. When added at the larval diet, harmine induced a strong reduction of larvae weight, cannibalism between larvae, in addition to significant mortality. On the other hand, it caused a remarkable development disruption, manifested by both delay and reduction of pupation and adult emergence. Using spectrophotometric assays, we have shown that harmine ingestion provoked a severe reduction in protein, glycogen and lipid contents. Beside, when larvae fed harmine, the activity of the digestive enzyme ?-amylase was strongly reduced. In conclusion, our experiments clearly show the susceptibility of P. interpunctella to harmine ingestion revealing the potent bioinsecticidal effect of harmine. PMID:23961164

  2. Neutrophil-activating protein mediates adhesion of Helicobacter pylori to sulfated carbohydrates on high-molecular-weight salivary mucin.

    PubMed

    Namavar, F; Sparrius, M; Veerman, E C; Appelmelk, B J; Vandenbroucke-Grauls, C M

    1998-02-01

    The in vitro binding of surface-exposed material and outer membrane proteins of Helicobacter pylori to high-molecular-weight salivary mucin was studied. We identified a 16-kDa surface protein which adhered to high-molecular-weight salivary mucin. This protein binds specifically to sulfated oligosaccharide structures such as sulfo-Lewis a, sulfogalactose and sulfo-N-acetyl-glucosamine on mucin. Sequence analysis of the protein proved that it was identical to the N-terminal amino acid sequence of neutrophil-activating protein. Moreover, this adhesin was able to bind to Lewis x blood group antigen. PMID:9453593

  3. Responses of Lipolysis and Salivary Cortisol to Food Intake and Physical Activity in Lean and Obese Children

    Microsoft Academic Search

    A. M. Hershberger; M. R. MCCAMMON; J. P. GARRY; M. T. MAHAR; R. C. HICKNER

    2004-01-01

    This investigation was conducted to determine whether there were differences in lipolytic responses to feeding and physical activity between lean (LN) and obese (OB) children, and if these responses were related to cortisol. Fourteen LN and 11 OB children participated in this study of abdominal lipolysis and salivary cortisol response to breakfast and lunch with an intervening exercise session. Calculated

  4. Levels of salivary lysozyme, lactoperoxidase, and lactoferrin in diabetic hamsters.

    PubMed Central

    Muratsu, K; Morioka, T

    1985-01-01

    In an attempt to clarify the mechanism(s) of increased susceptibility to oral infection in diabetics, we examined the levels of salivary antibacterial factors, including lysozyme, lactoperoxidase, and lactoferrin, in diabetic hamsters whose condition was induced with streptozotocin. Saliva was collected from these hamsters periodically for 19 weeks after the administration of streptozotocin. Diabetes persisted with significant hyperglycemia throughout the experiment after a single injection of streptozotocin. There was no significant difference between groups in the amount of saliva secreted. In diabetic hamsters, lysozyme activity decreased by 56% and lactoperoxidase activity decreased by 53% compared with the control hamsters 19 weeks after the administration of streptozotocin. There was no significant difference between groups in the amount of salivary lactoferrin. However, the ratio of lactoferrin to total protein increased to approximately double the amount of that of the control hamsters. Insulin treatment had a significant effect on lysozyme and lactoperoxidase activity, recovering 73 and 74% those of the controls, respectively, and the ratio of lactoferrin to total salivary protein reverted to normal values. Growth inhibition of Lactobacillus plantarum ATCC 8014 with whole saliva and amylase activity significantly decreased in diabetic hamsters. The position of each protein band of whole saliva on sodium dodecyl sulfate-polyacrylamide gel electrophoresis was almost the same for control and diabetic hamsters; however, there was some variability in band intensity. Images PMID:2580790

  5. Salt-dependent thermo-reversible ?-amylase: cloning and characterization of halophilic ?-amylase from moderately halophilic bacterium, Kocuria varians

    Microsoft Academic Search

    Rui Yamaguchi; Hiroko Tokunaga; Matsujiro Ishibashi; Tsutomu Arakawa; Masao Tokunaga

    2011-01-01

    A moderately halophilic bacterium, Kocuria varians, was found to produce active ?-amylase (K. varians ?-amylase (KVA)). We have observed at least six different forms of ?-amylase secreted by this bacterium into the culture\\u000a medium. Characterization of these KVA forms and cloning of the corresponding gene revealed that KVA comprises pre-pro-precursor\\u000a form of ?-amylase catalytic domain followed by the tandem repeats,

  6. Influence of chewing force on salivary stress markers as indicator of mental stress.

    PubMed

    Soeda, R; Tasaka, A; Sakurai, K

    2012-04-01

    The aim of this study was to investigate the influence of chewing force on salivary stress markers (alpha-amylase activity, salivary cortisol level and secretory immunoglobulin A secretion rate) as indicators of mental stress. Participants comprised 20 healthy men. The first set of saliva specimens (S1) was collected at immediately after a 20-min rest to evaluate stress markers. As stress loading, the participants were required to perform arithmetic calculations for 20 min, after which the second set of saliva specimens (S2) was collected. Each participant was then required to chew a piece of tasteless gum for 10 min, after which the third set of saliva specimens (S3) was collected. After a 20-min rest, the fourth set of saliva specimens (S4) was collected. Weak, habitual and strong chewing forces were assigned. Change rates of stress markers between S2 and S3, and S2 and S4 were calculated. A significant difference was observed in the change rate of cortisol levels between S2 and S3. Cortisol level decreased more under strong chewing than under weak chewing. No significant differences were observed in the change rate of amylase activity or s-IgA secretion rate among the three chewing forces. The results suggest that differences in chewing force influence the salivary cortisol level of the three stress markers, and that a strong chewing force induces a greater reduction in mental stress than a weak one. PMID:22040229

  7. Profiles and ?-amylase inhibition activity of proanthocyanidins in unripe Manilkara zapota (chiku).

    PubMed

    Wang, Hongyu; Liu, Tingting; Song, Lixia; Huang, Dejian

    2012-03-28

    Proanthocyanidins in unripe Manilkara zapota (chiku) were isolated using solvent extraction followed by Sephadex LH-20 fractionation with a yield of 0.9%. HPLC analysis using a diol column revealed well-resolved oligomers ranging from dimer to hexamer. The majority of the proanthocyanidins are composed of higher-degree oligomers appearing as one large peak in the chromatogram. Analysis of the proanthocyanidins using LC/MS showed that (epi)gallocatechins were the dominant extension unit in the proanthocyanidins. In agreement with this result, thiolysis treatment of the proanthocyanidins using mercaptoacetic acid produced thioether derivatives of (epi)gallocatechins as the major product and (epi)gallocatechin gallate derivatives as the minor product. The mean of the degree of polymerization was estimated to be 9.0. From MALDI-TOF MS, B-type gallocatechin oligomers up to decamer could be detected. The unripe chiku proanthocyanidins are thus good starting material for preparation of (epi)gallocatechin derivatives. The proanthocyanidins was shown to inhibit ?-amylase with an IC(50) value of 4.2 ± 0.2 ?g/mL and inhibit ?-glucosidase with an IC(50) of 16.6 ± 0.3 ?g/mL. PMID:22394060

  8. Salivary Glands

    MedlinePLUS

    ... parotid, submandibular, and sublingual glands. They all secrete saliva into your mouth, the parotid through tubes that drain saliva, called salivary ducts, near your upper teeth, submandibular ...

  9. Expression and Localization of ?-amylase in the Submandibular and Sublingual Glands of Mice

    PubMed Central

    Yamagishi, Ryoko; Wakayama, Tomohiko; Nakata, Hiroki; Adthapanyawanich, Kannika; Kumchantuek, Tewarat; Yamamoto, Miyuki; Iseki, Shoichi

    2014-01-01

    In the major salivary glands of mice, acinar cells in the parotid gland (PG) are known to be the main site for the production of the digestive enzyme ?-amylase, whereas ?-amylase production in the submandibular gland (SMG) and sublingual gland (SLG), as well as the cell types responsible for ?-amylase production, has been less firmly established. To clarify this issue, we examined the expression and localization of both the mRNA and protein of ?-amylase in the major salivary glands of male and female mice by quantitative and histochemical methods. ?-amylase mRNA levels were higher in the order of PG, SMG, and SLG. No sexual difference was observed in ?-amylase mRNA levels in the PG and SLG, whereas ?-amylase mRNA levels in the female SMG were approximately 30% those in the male SMG. Using in situ hybridization and immunohistochemistry, signals for ?-amylase mRNA and protein were found to be strongly positive in acinar cells of the PG, serous demilune cells of the SLG, and granular convoluted tubule (GCT) cells of the male SMG, weakly positive in seromucous acinar cells of the male and female SMG, and negative in mucous acinar cells of the SLG. These results clarified that ?-amylase is produced mainly by GCT cells and partly by acinar cells in the SMG, whereas it is produced exclusively by serous demilune cells in the SLG of mice. PMID:25320406

  10. Responses of lipolysis and salivary cortisol to food intake and physical activity in lean and obese children.

    PubMed

    Hershberger, A M; McCammon, M R; Garry, J P; Mahar, M T; Hickner, R C

    2004-09-01

    This investigation was conducted to determine whether there were differences in lipolytic responses to feeding and physical activity between lean (LN) and obese (OB) children, and if these responses were related to cortisol. Fourteen LN and 11 OB children participated in this study of abdominal lipolysis and salivary cortisol response to breakfast and lunch with an intervening exercise session. Calculated fasting glycerol release was lower in OB than LN (0.645 +/- 0.06 vs. 0.942 +/- 0.11 micromol/ml; P < 0.05). Fasting adipose tissue nutritive flow was lower in OB than in LN subjects, but responses to feeding and exercise were not different. Breakfast elicited a decrease in interstitial glycerol concentration in LN (-33%; P < 0.05), but not in OB (-5%), children, although decreases in glycerol concentration in response to lunch were similar (LN, -41%; OB, -36%). An interaction was evident in the salivary cortisol response to breakfast (LN, no change; OB, increase) and exercise (LN, no change; OB, decrease), but there were no group differences in response to lunch. Alterations in salivary cortisol and lipolysis were not related. These data suggest that salivary cortisol and lipolytic responses are not necessarily linked, but are altered in obesity. Furthermore, prior exercise may improve the antilipolytic response to a meal in OB children. PMID:15356083

  11. Distinct regulation by lipopolysaccharides of the expression of interleukin-1? by murine macrophages and salivary glands.

    PubMed

    Seil, M; El Ouaaliti, M; Abdou Foumekoye, S; Pochet, S; Dehaye, J P

    2012-02-01

    The regulation of interleukin (IL)-1 expression and secretion by salivary glands and macrophages in response to lipopolysaccharides (LPS) was compared. In wild-type mice, injection of LPS significantly decreased the volume of saliva stimulated by pilocarpine and increased its protein and amylase concentration. It did not modify the salivary concentration of IL-1?. The cytokine was expressed by submandibular acini and ducts. Macrophages also expressed IL-1? but at lower concentration than salivary glands. The pre-incubation of macrophages with LPS increased the phosphorylation of I?B and the expression of IL-1?. Adenosine triphosphate also promoted the secretion of the cytokine by these cells. These responses were absent in submandibular gland cells. These glands expressed CD14, TLR4 and MyD88. P2X(7)-KO mice secreted a lower volume of saliva which contained less proteins and amylase. In conclusion, IL-1? is constitutively expressed by submandibular glands and its secretion is not regulated by a P2X(7) agonist. In these cells, LPS do not activate the nuclear factor-?B-pro-IL-1? axis in spite of the expression of the proteins involved in their recognition. PMID:20682589

  12. Effect of laser phototherapy on enzymatic activity of salivary glands of hamsters treated with 5-Fluorouracil.

    PubMed

    Campos, Luana; Nicolau, José; Arana-Chavez, Victor E; Simões, Alyne

    2014-01-01

    The chemotherapeutic agent 5-Fluorouracil (5-FU) can induce salivary gland hypofunction (SGH); however, previous studies did not reach final conclusions on the influence of this drug on glandular tissue. Thus, the aim of this study was to investigate the effect of 5-FU on submandibular (SMs) and sublingual glands (SLs), as well as, the effect of laser phototherapy (LPT) on SGH induced by 5-FU. Eighty-five hamsters were divided into three groups: control (C), chemotherapy (CT) and laser (L), and the SGH was induced by two injections of 5-FU in groups CT and L. The irradiation was performed using a diode (?780 nm/20 mW/5 J cm(-2)/0.2 J and 10 s per point/spot size of 0.04 cm(2)) and applied daily. On the euthanasia day, SMs and SLs were removed and biochemical analyses were carried out. The lactate dehydrogenase activity was increased in group CT when compared with group C for SLs and SMs (P < 0.05). In addition, the peroxidase and catalase activities were increased and superoxide dismutase was decreased by 5-FU (P < 0.05). However, LPT appears to be a protective mechanism against oxidative stress, tending to alter the activity of these antioxidant enzymes, suggesting LPT as a promising therapy to modulate the 5-FU harmful effect. PMID:24172058

  13. Diosgenin from Dioscorea bulbifera: Novel Hit for Treatment of Type II Diabetes Mellitus with Inhibitory Activity against ?-Amylase and ?-Glucosidase

    PubMed Central

    Ghosh, Sougata; More, Piyush; Derle, Abhishek; Patil, Ajay B.; Markad, Pramod; Asok, Adersh; Kumbhar, Navanath; Shaikh, Mahemud L.; Ramanamurthy, Boppana; Shinde, Vaishali S.; Dhavale, Dilip D.; Chopade, Balu A.

    2014-01-01

    Diabetes mellitus is a multifactorial metabolic disease characterized by post-prandial hyperglycemia (PPHG). ?-amylase and ?-glucosidase inhibitors aim to explore novel therapeutic agents. Herein we report the promises of Dioscorea bulbifera and its bioactive principle, diosgenin as novel ?-amylase and ?-glucosidase inhibitor. Among petroleum ether, ethyl acetate, methanol and 70% ethanol (v/v) extracts of bulbs of D. bulbifera, ethyl acetate extract showed highest inhibition upto 72.06 ± 0.51% and 82.64 ± 2.32% against ?-amylase and ?-glucosidase respectively. GC-TOF-MS analysis of ethyl acetate extract indicated presence of high diosgenin content. Diosgenin was isolated and identified by FTIR, 1H NMR and 13C NMR and confirmed by HPLC which showed an ?-amylase and ?-glucosidase inhibition upto 70.94 ± 1.24% and 81.71 ± 3.39%, respectively. Kinetic studies confirmed the uncompetitive mode of binding of diosgenin to ?-amylase indicated by lowering of both Km and Vm. Interaction studies revealed the quenching of intrinsic fluorescence of ?-amylase in presence of diosgenin. Similarly, circular dichroism spectrometry showed diminished negative humped peaks at 208 nm and 222 nm. Molecular docking indicated hydrogen bonding between carboxyl group of Asp300, while hydrophobic interactions between Tyr62, Trp58, Trp59, Val163, His305 and Gln63 residues of ?-amylase. Diosgenin interacted with two catalytic residues (Asp352 and Glu411) from ?-glucosidase. This is the first report of its kind that provides an intense scientific rationale for use of diosgenin as novel drug candidate for type II diabetes mellitus. PMID:25216353

  14. Cyclic AMP regulation of calcium mobilization and amylase release from isolated permeabilized rat parotid cells.

    PubMed

    Rubin, R P; Adolf, M A

    1994-02-01

    This study examined the mechanistic basis of the synergistic interaction between the cyclic AMP (cAMP) and phosphoinositide pathways in salivary amylase secretion. cAMP produced a concentration-dependent increase in Ca++ mobilization from saponin-permeabilized rat parotid acinar cells. A threshold concentration of cAMP (50 microM) significantly increased the peak Ca(++)-releasing activity of submaximal concentrations of inositol 1,4,5-trisphosphate (IP3) but did not augment the Ca++ mobilization induced by a maximal stimulating concentration of IP3 (30 microM). A maximal stimulating concentration of cAMP (500 microM) failed to modify the Ca++ releasing action of IP3. IP3-induced Ca++ release was also augmented by catalytic subunit of cAMP-dependent protein kinase. A specific protein kinase inhibitor reversed this effect. The cAMP-induced Ca++ release was blocked by ryanodine but not by heparin, by contrast with the IP3-induced Ca++ release. Thapsigargin only partially depressed the cAMP response but completely abolished the IP3 response. The amylase release elicited by fixed concentrations of Ca++ was not further enhanced by either cAMP or forskolin. Thus, unlike diacylglycerol, which decreases the Ca++ requirement for secretion by inducing activation of protein kinase C, cAMP appears to mediate salivary amylase secretion by regulating the sensitivity of parotid cells to the Ca++ mobilizing action of IP3. In addition, cAMP possesses a second action, i.e., directly eliciting Ca++ mobilization from an IP3-insensitive pool. PMID:7509390

  15. Lundep, a Sand Fly Salivary Endonuclease Increases Leishmania Parasite Survival in Neutrophils and Inhibits XIIa Contact Activation in Human Plasma

    PubMed Central

    Chagas, Andrezza C.; Oliveira, Fabiano; Debrabant, Alain; Valenzuela, Jesus G.; Ribeiro, José M. C.; Calvo, Eric

    2014-01-01

    Neutrophils are the host's first line of defense against infections, and their extracellular traps (NET) were recently shown to kill Leishmania parasites. Here we report a NET-destroying molecule (Lundep) from the salivary glands of Lutzomyia longipalpis. Previous analysis of the sialotranscriptome of Lu. longipalpis showed the potential presence of an endonuclease. Indeed, not only was the cloned cDNA (Lundep) shown to encode a highly active ss- and dsDNAse, but also the same activity was demonstrated to be secreted by salivary glands of female Lu. longipalpis. Lundep hydrolyzes both ss- and dsDNA with little sequence specificity with a calculated DNase activity of 300000 Kunitz units per mg of protein. Disruption of PMA (phorbol 12 myristate 13 acetate)- or parasite-induced NETs by treatment with recombinant Lundep or salivary gland homogenates increases parasite survival in neutrophils. Furthermore, co-injection of recombinant Lundep with metacyclic promastigotes significantly exacerbates Leishmania infection in mice when compared with PBS alone or inactive (mutagenized) Lundep. We hypothesize that Lundep helps the parasite to establish an infection by allowing it to escape from the leishmanicidal activity of NETs early after inoculation. Lundep may also assist blood meal intake by lowering the local viscosity caused by the release of host DNA and as an anticoagulant by inhibiting the intrinsic pathway of coagulation. PMID:24516388

  16. Expression of beta-amylase from alfalfa taproots.

    PubMed

    Gana, J A; Kalengamaliro, N E; Cunningham, S M; Volenec, J J

    1998-12-01

    Alfalfa (Medicago sativa L.) roots contain large quantities of beta-amylase, but little is known about its role in vivo. We studied this by isolating a beta-amylase cDNA and by examining signals that affect its expression. The beta-amylase cDNA encoded a 55.95-kD polypeptide with a deduced amino acid sequence showing high similarity to other plant beta-amylases. Starch concentrations, beta-amylase activities, and beta-amylase mRNA levels were measured in roots of alfalfa after defoliation, in suspension-cultured cells incubated in sucrose-rich or -deprived media, and in roots of cold-acclimated germ plasms. Starch levels, beta-amylase activities, and beta-amylase transcripts were reduced significantly in roots of defoliated plants and in sucrose-deprived cell cultures. beta-Amylase transcript was high in roots of intact plants but could not be detected 2 to 8 d after defoliation. beta-Amylase transcript levels increased in roots between September and October and then declined 10-fold in November and December after shoots were killed by frost. Alfalfa roots contain greater beta-amylase transcript levels compared with roots of sweetclover (Melilotus officinalis L.), red clover (Trifolium pratense L.), and birdsfoot trefoil (Lotus corniculatus L.). Southern analysis indicated that beta-amylase is present as a multigene family in alfalfa. Our results show no clear association between beta-amylase activity or transcript abundance and starch hydrolysis in alfalfa roots. The great abundance of beta-amylase and its unexpected patterns of gene expression and protein accumulation support our current belief that this protein serves a storage function in roots of this perennial species. PMID:9847126

  17. Salivary gland acinar cells regenerate functional glandular structures in modified hydrogels

    NASA Astrophysics Data System (ADS)

    Pradhan, Swati

    Xerostomia, a condition resulting from irradiation of the head and neck, affects over 40,000 cancer patients each year in the United States. Direct radiation damage of the acinar cells that secrete fluid and protein results in salivary gland hypofunction. Present medical management for xerostomia for patients treated for upper respiratory cancer is largely ineffective. Patients who have survived their terminal diagnosis are often left with a diminished quality of life and are unable to enjoy the simple pleasures of eating and drinking. This project aims to ultimately reduce human suffering by developing a functional implantable artificial salivary gland. The goal was to create an extracellular matrix (ECM) modified hyaluronic acid (HA) based hydrogel culture system that allows for the growth and differentiation of salivary acinar cells into functional acini-like structures capable of secreting large amounts of protein and fluid unidirectionally and to ultimately engineer a functional artificial salivary gland that can be implanted into an animal model. A tissue collection protocol was established and salivary gland tissue was obtained from patients undergoing head and neck surgery. The tissue specimen was assessed by histology and immunohistochemistry to establish the phenotype of normal salivary gland cells including the native basement membranes. Hematoxylin and eosin staining confirmed normal glandular tissue structures including intercalated ducts, striated ducts and acini. alpha-Amylase and periodic acid schiff stain, used for structures with a high proportion of carbohydrate macromolecules, preferentially stained acinar cells in the tissue. Intercalated and striated duct structures were identified using cytokeratins 19 and 7 staining. Myoepithelial cells positive for cytokeratin 14 were found wrapped around the serous and mucous acini. Tight junction components including ZO-1 and E-cadherin were present between both ductal and acinar cells. Ductal and acinar cells were identified in cultured cells from dispersed tissue. Biomarker studies with the salivary enzyme, alpha-amylase, and tight junction proteins, such as zonula occludens-1 and E-cadherin, confirmed the phenotype of these cells. Strong staining for laminin and perlecan/HSPG2 were noted in basement membranes and perlecan also was secreted and organized by cultured acinar populations, which formed lobular structures that mimicked intact glands when cultured on Matrigel(TM) or a bioactive peptide derived from domain IV of perlecan (PlnDIV). On either matrix, large acini-like lobular structures grew and formed connections between the lobes. alpha-Amylase secretion was confirmed by staining and activity assay. Biomarkers including tight junction protein E-cadherin and water channel protein, aquaporin 5 (AQP5) found in tissue, were expressed in cultured acinar cells. Cells cultured on Matrigel(TM) or PlnDIV peptide organized stress fibers and activated focal adhesion kinase (FAK). HA, a natural polysaccharide and a major component of the ECM, can be used to generate soft and pliable hydrogels. A culture system consisting of HA hydrogel and PlnDIV peptide was used to generate a 2.5D culture system. Acinar cells cultured on these hydrogels self-assembled into lobular structures and expressed tight junction components such as ZO-1. Acini-like structures were stained for the presence of alpha-amylase. Live/dead staining revealed the presence of apoptotic cells in the center of the acini-like structures, indicative of lumen formation. The functionality of these acini-like structures was studied by stimulating them with neurotransmitters to enhance their fluid and protein production. Acini-like structures treated with norepinephrine and isoproterenol showed increased granule formation as observed by phase contrast microscopy and alpha-amylase staining in the structures. Lobular structures on hydrogels were treated with acetylcholine to increase fluid production. The increase in intracellular calcium due to the activation of the M3 muscarinic receptor via binding to ac

  18. Hair cortisol levels as a retrospective marker of hypothalamic-pituitary axis activity throughout pregnancy: comparison to salivary cortisol.

    PubMed

    D'Anna-Hernandez, Kimberly L; Ross, Randal G; Natvig, Crystal L; Laudenslager, Mark L

    2011-08-01

    Maternal stress during pregnancy is associated with negative maternal/child outcomes. One potential biomarker of the maternal stress response is cortisol, a product of activity of the hypothalamic-pituitary-adrenal axis. This study evaluated cortisol levels in hair throughout pregnancy as a marker of total cortisol release. Cortisol levels in hair have been shown to be easily quantifiable and may be representative of total cortisol release more than single saliva or serum measures. Hair cortisol provides a simple way to monitor total cortisol release over an extended period of time. Hair cortisol levels were determined from each trimester (15, 26 and 36 weeks gestation) and 3 months postpartum. Hair cortisol levels were compared to diurnal salivary cortisol collected over 3 days (3 times/day) at 14, 18, 23, 29, and 34 weeks gestational age and 6 weeks postpartum from 21 pregnant women. Both salivary and hair cortisol levels rose during pregnancy as expected. Hair cortisol and diurnal salivary cortisol area under the curve with respect to ground (AUCg) were also correlated throughout pregnancy. Levels of cortisol in hair are a valid and useful tool to measure long-term cortisol activity. Hair cortisol avoids methodological problems associated with collection other cortisol measures such as plasma, urine, or saliva and is a reliable metric of HPA activity throughout pregnancy reflecting total cortisol release over an extended period. PMID:21397617

  19. A study into salivary-based measurement of human stress subjected to ellestad stress test protocol

    Microsoft Academic Search

    Y. K. Lee; A. Za'aba; N. K. Madzhi; A. Ahmad

    2009-01-01

    Previous works on the effects of salivary alpha amylase in respond to various stressors report encouraging findings on it being a good indicator of stress. Ellestad protocol is a clinical procedure to screen for coronary artery disease by introducing exercise induced physical stress. If a salivary based biomarker profile in accordance to a stress test protocol could be established, the

  20. Cortisol and Children's Adjustment: The Moderating Role of Sympathetic Nervous System Activity

    ERIC Educational Resources Information Center

    El-Sheikh, Mona; Erath, Stephen A.; Buckhalt, Joseph A.; Granger, Douglas A.; Mize, Jacquelyn

    2008-01-01

    We examined relations among cortisol, markers of sympathetic nervous system (SNS) activity (including salivary alpha-amylase and skin conductance level), and children's adjustment. We also tested the Bauer et al. ("Journal of Developmental and Behavioral Pediatrics," 23(2), 102-113, 2002) hypothesis that interactions between the SNS and cortisol…

  1. Tsetse Salivary Gland Proteins 1 and 2 Are High Affinity Nucleic Acid Binding Proteins with Residual Nuclease Activity

    PubMed Central

    Caljon, Guy; Ridder, Karin De; Stijlemans, Benoît; Coosemans, Marc; Magez, Stefan; De Baetselier, Patrick; Van Den Abbeele, Jan

    2012-01-01

    Analysis of the tsetse fly salivary gland EST database revealed the presence of a highly enriched cluster of putative endonuclease genes, including tsal1 and tsal2. Tsal proteins are the major components of tsetse fly (G. morsitans morsitans) saliva where they are present as monomers as well as high molecular weight complexes with other saliva proteins. We demonstrate that the recombinant tsetse salivary gland proteins 1&2 (Tsal1&2) display DNA/RNA non-specific, high affinity nucleic acid binding with KD values in the low nanomolar range and a non-exclusive preference for duplex. These Tsal proteins exert only a residual nuclease activity with a preference for dsDNA in a broad pH range. Knockdown of Tsal expression by in vivo RNA interference in the tsetse fly revealed a partially impaired blood digestion phenotype as evidenced by higher gut nucleic acid, hematin and protein contents. PMID:23110062

  2. Starch Phosphorylase Inhibitor Is ?-Amylase 1

    PubMed Central

    Pan, Shu-Mei; Chang, Tsung-Chain; Juang, Rong-Huay; Su, Jong-Ching

    1988-01-01

    The proteinaceous noncompetitive inhibitor of starch phosphorylase isolated from the root of sweet potato (Ipomoea batatas [L.] Lam.) (TC Chang, JC Su 1986 Plant Physiol 80: 534-538) has been identified as a ?-amylase. The starch phosphorylase inhibitor and ?-amylase activities copurified to give a protein indistinguishable from commercial ?-amylase by electrophoretic and immunological methods, and the two activities showed parallel responses in pH, temperature, and inhibitor sensitivity tests. The amylolytic pattern of the inhibitor corresponded to that of ?-amylase and its inhibitory effect toward starch phosphorylase was due to neither deprivation of starch, the primer for the phosphorylase assay, nor the inhibitory effect of amylolytic products. Images Fig. 2 Fig. 3 PMID:16666436

  3. Diurnal variation of amylase secretion is coupled with alterations of ?-adrenoceptors in the rat parotid gland

    Microsoft Academic Search

    Y. Ishikawa; I. Amano; C. Chen; H. Ishida

    1992-01-01

    Summary  Diurnal changes in the neurotransmitter receptors are important for studying the receptor function in neurophysiology. The\\u000a purpose of this study is to gain an insight into the regulatory mechanisms of the diurnal variation of amylase secretion.\\u000a Rat salivary amylase levels showed a diurnal variation with two peaks, a marked peak at 13 h and a lesser peak at 21 h.

  4. Inhibition of Sunn Pest, Eurygaster integriceps, ?-Amylases by ?-Amylase Inhibitors (T-?AI) from Triticale

    PubMed Central

    Mehrabadi, Mohammad; Bandani, Ali R.; Saadati, Fatemeh

    2010-01-01

    The effect of triticale ?-amylases inhibitors on starch hydrolysis catalyzed by the Sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae) midgut amylases was examined. Biochemical studgawies showed that inhibitors from Triticale (a hybrid of wheat and rye) had inhibitiory effects on E. integriceps ?-amylases. The effects of the triticale ?-amylase inhibitor (T-?AI) on ?-amylase of E. integriceps showed a dose dependent manner of inhibition, e.g. less inhibition of enzyme activity (around 10%) with a lower dose (0.25 mg protein) and high inhibition of enzyme activity (around 80%) when a high dose of inhibitor was used (1.5 mg protein). The enzyme kinetic studies using Michaelis-Menten and Lineweaver-Burk equations showed the Km remained constant (0.58%) but the maximum velocity (Vmax) decreased in the presence of a crude extract of Triticale inhibitors, indicating mixed inhibition. The temperature giving 50% inactivation of enzyme (T50) during a 30-min incubation at pH 7.0 was 73° C. The maximum inhibitory activity was achieved at 35° C and pH 5.0. Gel assays showed the meaningful inhibition of E. integriceps ?-amylases by various concentrations of Triticale inhibitors. Based on the data presented in this study, it could be said that the T-?AI has good inhibitory activity on E. integriceps gut ?-amylase. PMID:21062146

  5. Chromosome Puff Activity and Protein Synthesis in Larval Salivary Glands of Drosophila melanogaster

    Microsoft Academic Search

    Gunter Korge

    1975-01-01

    Secretion proteins from larval salivary glands of Drosophila melanogaster were analyzed with acrylamide gel electrophoresis. Four fractions were found; three showed electrophoretic variants in different wild-type stocks. Crossbreeding and cytogenetic techniques were used to localize the genes responsible for the two main fractions: The gene for fraction 3 was found to lie within a segment of the third chromosome which

  6. Patterns of puffing activity in the salivary gland chromosomes of Drosophila

    Microsoft Academic Search

    Michael Ashburner

    1970-01-01

    Exposure of Drosophila melanogaster larvae to high temperature for short periods of time results in marked changes in the puffing patterns of salivary gland chromosomes. Temperature shock “induces” puffing at 9 specific loci; this pattern of induced puffs shows little developmental specificity and is similar in three strains of D. melanogaster (including the mutant lethal giant-larvae) and in D. simulans.

  7. Parallel Changes in Puffing Activity and Patterns of Protein Synthesis in Salivary Glands of Drosophila

    Microsoft Academic Search

    Michael Lewis; Pieter J. Helmsing; Michael Ashburner

    1975-01-01

    The changes in protein synthesis of salivary glands of Drosophila resulting from a brief exposure to 37 degrees have been analyzed on sodium dodecyl sulfate-acrylamide gels. In D. melanogaster and D. hydei this treatment induces nine and six new puffs, respectively, in the polytene chromosomes. After 20 min treatment seven new proteins are synthesized by the glands of D. melanogaster

  8. Effects of tea polyphenols on the activities of ?-amylase, pepsin, trypsin and lipase

    Microsoft Academic Search

    Qiang He; Yuanping Lv; Kai Yao

    2007-01-01

    Tea polyphenols (TP) possess many beneficial properties, such as reducing the risk of cancer and heart diseases, and acting as natural antioxidants for the food industry. At the same time, tea polyphenols might inhibit digestive enzymes and reduce food digestibility. To explore this possible antinutritional property, the effects of tea polyphenols on the activity of four typical digestive enzymes were

  9. Effect of genetic polymorphisms in CA6 gene on the expression and catalytic activity of human salivary carbonic anhydrase VI.

    PubMed

    Aidar, M; Marques, Rocha; Valjakka, J; Mononen, N; Lehtimäki, T; Parkkila, S; de Souza, A P; Line, S R Peres

    2013-01-01

    Carbonic anhydrase isoenzyme VI (CA VI) plays an important role in the homeostasis of oral tissues participating in the processes of taste, protection of dental tissues against the loss of minerals, caries, and possibly in the formation of dental calculus in periodontal disease. This study aimed to verify the correlation between changes in the expression and activity of human salivary carbonic anhydrase VI and genetic polymorphisms in its gene (CA6). The study population consisted of 182 healthy volunteers (female and male, aged 18-22). Samples of total saliva were assayed for CA VI concentrations using a specific time-resolved immunofluorometric assay. CA VI catalytic activity was detected by a modified protocol of Kotwica et al. [J Physiol Pharmacol 2006;57(suppl 8):107-123], adapted to CA VI in saliva. Samples of genomic DNA were genotyped for polymorphisms rs2274327 (C/T), rs2274328 (A/C) and rs2274333 (A/G) by TaqMan® SNP Genotyping Assays. The concentration and catalytic activity of the salivary CA VI obtained for the different genotypes were analyzed using the Kruskal-Wallis nonparametric test and the Dunn test. The results showed that individuals with TT genotype (rs2274327) had significantly lower CA VI concentrations than the individuals with genotypes CT or CC (p < 0.05). There was also an association between polymorphism rs2274333 and salivary CA VI concentrations. There were no associations between the three polymorphisms analyzed and variations in CA VI activity. Our results suggest that polymorphisms in the CA6 gene are associated with the concentrations of secreted CA VI. PMID:23652931

  10. Crystallization and X-ray crystallographic studies of an inhibitor from rye (Secale cereale) active against Acanthoscelides obtectus and Zabrotes subfasciatus alpha-amylases.

    PubMed

    Iulek, Jorge; Slivinskic, Christiane Trevisan; Silva, Márcio

    2004-02-01

    Crystals of a new inhibitor present in rye seeds active against alpha-amylases from crop pests Acanthoscelides obtectus and Zabrotes subfasciatus have been obtained. A native dataset was collected at 2.21 A resolution with 99.3% completeness at CPr beamline at LNLS. The crystals belong to the trigonal system, space group P3(1)21 with a=b=78.21 A, and c=59.61 A. The crystal calculated solvent content is compatible with one dimer per asymmetric unit. PMID:14965283

  11. Novel Family of Insect Salivary Inhibitors Blocks Contact Pathway Activation by Binding to Polyphosphate, Heparin, and Dextran Sulfate

    PubMed Central

    Alvarenga, Patricia H.; Xu, Xueqing; Oliveira, Fabiano; Chagas, Andrezza C.; Nascimento, Clarissa R.; Francischetti, Ivo M.B.; Juliano, Maria A.; Juliano, Luiz; Scharfstein, Julio; Valenzuela, Jesus G.; Ribeiro, José M.C.; Andersen, John F.

    2014-01-01

    Objective Polyphosphate and heparin are anionic polymers released by activated mast cells and platelets that are known to stimulate the contact pathway of coagulation. These polymers promote both the autoactivation of factor XII and the assembly of complexes containing factor XI, prekallikrein, and high-molecular-weight kininogen. We are searching for salivary proteins from blood-feeding insects that counteract the effect of procoagulant and proinflammatory factors in the host, including elements of the contact pathway. Approach and Results Here, we evaluate the ability of the sand fly salivary proteins, PdSP15a and PdSP15b, to inhibit the contact pathway by disrupting binding of its components to anionic polymers. We attempt to demonstrate binding of the proteins to polyphosphate, heparin, and dextran sulfate. We also evaluate the effect of this binding on contact pathway reactions. We also set out to determine the x-ray crystal structure of PdSP15b and examine the determinants of relevant molecular interactions. Both proteins bind polyphosphate, heparin, and dextran sulfate with high affinity. Through this mechanism they inhibit the autoactivation of factor XII and factor XI, the reciprocal activation of factor XII and prekallikrein, the activation of factor XI by thrombin and factor XIIa, the cleavage of high-molecular-weight kininogen in plasma, and plasma extravasation induced by polyphosphate. The crystal structure of PdSP15b contains an amphipathic helix studded with basic side chains that forms the likely interaction surface. Conclusions The results of these studies indicate that the binding of anionic polymers by salivary proteins is used by blood feeders as an antihemostatic/anti-inflammatory mechanism. PMID:24092749

  12. High amylase content of neoplastic pleural and pericardial effusion probably secondary to amylase producing tumor cells: Report of 2 cases

    Microsoft Academic Search

    N. Satz; R. Miinch; U. Kuhlmann; G. Pedio; D. Gut; P. Pei; R. W. Ammann

    1983-01-01

    Summary We report two cases of malignant pleural and pericardial effusion respectively secondary to bronchogenic carcinomas. In both effusions, a significant elevation of the Salivary-type-amylase fraction was found, while the corresponding values were normal in serum and urine. Electronmicroscopy of the malignant tumor cells from the pleural effusion showed typical electron-dense granules, suggesting zymogen granules. It is concluded that the

  13. Growth Factors Polymerized Within Fibrin Hydrogel Promote Amylase Production in Parotid Cells

    PubMed Central

    McCall, Andrew D.; Nelson, Joel W.; Leigh, Noel J.; Duffey, Michael E.; Lei, Pedro; Andreadis, Stelios T.

    2013-01-01

    Salivary gland cell differentiation has been a recurring challenge for researchers as primary salivary cells show a loss of phenotype in culture. Particularly, parotid cells show a marked decrease in amylase expression, the loss of tight junction organization and proper cell function. Previously, Matrigel has been used successfully as an extracellular matrix; however, it is not practical for in vivo applications as it is tumorigenic. An alternative method could rely on the use of fibrin hydrogel (FH), which has been used extensively in biomedical engineering applications ranging from cardiovascular tissue engineering to wound-healing experiments. Although several groups have examined the effects of a three-dimensional (3D) environment on salivary cell cultures, little is known about the effects of FH on salivary cell cultures. The current study developed a 3D cell culture model to support parotid gland cell differentiation using a combination of FH and growth factor-reduced Matrigel (GFR-MG). Furthermore, FH polymerized with a combination of EGF and IGF-1 induced formation of 3D spheroids capable of amylase expression and an agonist-induced increase in the intracellular Ca2+ concentration ([Ca2+]i) in salivary cells. These studies represent an initial step toward the construction of an artificial salivary gland to restore salivary gland dysfunction. This is necessary to reduce xerostomia in patients with compromised salivary function. PMID:23594102

  14. Bone-muscle unit activity, salivary steroid hormones profile, and physical effort over a 3-week stage race.

    PubMed

    Grasso, D; Corsetti, R; Lanteri, P; Di Bernardo, C; Colombini, A; Graziani, R; Banfi, G; Lombardi, G

    2015-02-01

    Muscle traction and bone metabolism are functionally linked and co-regulated by a series of factors. Although a role for steroid hormones was hypothesized, a clear definition of the bone-muscle interconnection still lacks. To investigate this relationship, we studied bone metabolism, muscle activity, and salivary steroid hormones profile in relation with the physical effort across a cycling stage race, a model of effort in absence of load. Nine pro-cyclists were recruited; body weight and power output/energy expenditure were recorded. Diet was kept constant. Saliva was collected at days -1, 4, 8, 12, 14, 19, and 23; blood and urine were collected at days -1, 12, and 23. Salivary steroid hormones [cortisol, dehydroepiandrosterone (DHEA), testosterone, and estradiol], serum lactate dehydrogenase (LDH), aspartate aminotransferase (AST) and creatine kinase (CK) activities, plasma sclerostin, and urinary calcium and phosphorous were measured. Cortisol remained constant, testosterone decreased at day 4, and estradiol and DHEA firstly increased and then returned to basal levels. Hormone concentrations were not correlated with plasma volume shifts. LDH, CK, AST, sclerostin, and urinary calcium and phosphorous increased. DHEA and estradiol correlated with the physical effort and the bone-muscular markers. A relationship between muscle activity, in absence of load, and bone resorption emerged under a putative regulation by DHEA and estradiol. PMID:24433517

  15. ?-Amylases of the coffee berry borer ( Hypothenemus hampei) and their inhibition by two plant amylase inhibitors

    Microsoft Academic Search

    Arnubio Valencia; Alex E Bustillo; Gustavo E Ossa; Maarten J Chrispeels

    2000-01-01

    The adult coffee berry borer (Hypothenemus hampei Ferrari [Coleoptera: Scolytidae]), a major insect pest of coffee, has two major digestive ?-amylases that can be separated by isoelectric focusing. The ?-amylase activity has a broad pH optimum between 4.0 and 7.0. Using pH indicators, the pH of the midgut was determined to be between 4.5 and 5.2. At pH5.0, the coffee

  16. Production of Fungal Amylases Using Cheap, Readily Available Agriresidues, for Potential Application in Textile Industry

    PubMed Central

    Singh, Sanamdeep; Bali, Vrinda; Mangla, Jyoti

    2014-01-01

    The study aimed at isolation and screening of fungal amylase producer, optimization of solid state fermentation conditions for maximum amylase production by the best amylase producer, and characterization of the crude amylases, so produced. Aspergillus fumigatus NTCC1222 showed the highest amylase activity (164.1?U/mL) in secondary screening under SSF conditions and was selected for further studies. The test strain showed maximum amylase production (341.7?U/mL) and supernatant protein concentration (9.7?mg/mL) for incubation period (6 days), temperature (35°C), initial pH (6.0), nutrient salt solution as moistening agent, and beef extract as nitrogen source. Pomegranate peel produced maximum amylase activity, but wheat bran (only slightly lesser amylase activity as compared to that of pomegranate peel) was chosen for further studies, keeping in mind the seasonal availability of pomegranate peel. TLC confirmed the amylase produced to be ?-type and 60?kDa was the molecular weight of the partially purified amylase. The enzyme showed maximum enzyme activity at pH 6.0, temperature of 55°C, and incubation time of 60 minutes. UV (616.0?U/mL) and chemical (814.2?U/mL) mutation enhanced amylase activity as compared to wild test strain. The study indicates that Aspergillus fumigatus NTCC1222 can be an important source of amylase and the crude enzyme, hence obtained, can be cost effectively applied in multiple sections of textile wet processing. PMID:24527439

  17. Production of fungal amylases using cheap, readily available agriresidues, for potential application in textile industry.

    PubMed

    Singh, Shalini; Singh, Sanamdeep; Bali, Vrinda; Sharma, Lovleen; Mangla, Jyoti

    2014-01-01

    The study aimed at isolation and screening of fungal amylase producer, optimization of solid state fermentation conditions for maximum amylase production by the best amylase producer, and characterization of the crude amylases, so produced. Aspergillus fumigatus NTCC1222 showed the highest amylase activity (164.1 U/mL) in secondary screening under SSF conditions and was selected for further studies. The test strain showed maximum amylase production (341.7 U/mL) and supernatant protein concentration (9.7 mg/mL) for incubation period (6 days), temperature (35 °C), initial pH (6.0), nutrient salt solution as moistening agent, and beef extract as nitrogen source. Pomegranate peel produced maximum amylase activity, but wheat bran (only slightly lesser amylase activity as compared to that of pomegranate peel) was chosen for further studies, keeping in mind the seasonal availability of pomegranate peel. TLC confirmed the amylase produced to be ? -type and 60 kDa was the molecular weight of the partially purified amylase. The enzyme showed maximum enzyme activity at pH 6.0, temperature of 55 °C, and incubation time of 60 minutes. UV (616.0 U/mL) and chemical (814.2 U/mL) mutation enhanced amylase activity as compared to wild test strain. The study indicates that Aspergillus fumigatus NTCC1222 can be an important source of amylase and the crude enzyme, hence obtained, can be cost effectively applied in multiple sections of textile wet processing. PMID:24527439

  18. Purification and characterization of ?-Amylase from Miswak Salvadora persica

    PubMed Central

    2014-01-01

    Background The miswak (Salvadora persica) is a natural toothbrush. It is well known that very little information has been reported on enzymes in miswak as medicinal plant. Recently, we study peroxidase in miswak. In the present study, the main goal of this work is to purify and characterize ?-amylase from miswak. The second goal is to study the storage stability of ?-amylase in toothpaste. Method The purification method included chromatographaphy of miswak ?-amylase on DEAE-Sepharose column and Sephacryl S-200 column. Molecular weight was determined by gel filtration and SDS-PAGE. Results Five ?-amylases A1, A4a, A4b, A5a and A5b from miswak were purified and they had molecular weights of 14, 74, 16, 30 and 20 kDa, respectively. ?-Amylases had optimum pH from 6 to 8. Affinity of the substrates toward all enzymes was studied. Miswak ?-amylases A1, A4a, A4b, A5a and A5b had Km values for starch and glycogen of 3.7, 3.7, 7.1, 0.52, 4.3 mg/ml and 5.95, 5.9 4.16, 6.3, 6.49 mg/ml, respectively. The optimum temperature for five enzymes ranged 40°C- 60°C. Miswak ?-amylases were stable up to 40°C- 60°C after incubation for 30 min. Ca+2 activated all the miswak ?-amylases, while Ni2+, Co+2 and Zn+2 activated or inhibited some of these enzymes. The metal chelators, EDTA, sodium citrate and sodium oxalate had inhibitory effects on miswak ?-amylases. PMSF, p-HMB, DTNB and 1,10 phenanthroline caused inhibitory effect on ?-amylases. The analysis of hydrolytic products after starch hydrolysis by miswak ?-amylases on paper chromatography revealed that glucose, maltose, maltotriose and oligosaccharide were the major products. Crude miswak ?-amylase in the toothpaste retained 55% of its original activity after 10 months of storage at room temperature. Conclusions From these findings, ?-amylases from miswak can be considered as beneficial enzymes for pharmaceuticals. Therefore, we study the storage stability of the crude ?-amylase of miswak, which contained the five ?-amylases, in toothpaste. The enzyme in the toothpaste retained 55% of its original activity after 10 months of storage at room temperature. PMID:24690287

  19. Immobilization of ?-Amylase onto Luffa operculata Fibers

    PubMed Central

    Morais, Ricardo R.; Pascoal, Aline M.; Caramori, Samantha S.; Lopes, Flavio M.; Fernandes, Kátia F.

    2013-01-01

    A commercial amylase (amy) was immobilized by adsorption onto Luffa operculata fibers (LOFs). The derivative LOF-amy presented capacity to hydrolyze starch continuously and repeatedly for over three weeks, preserving more than 80% of the initial activity. This system hydrolyzed more than 97% of starch during 5?min, at room temperature. LOF-amy was capable to hydrolyze starch from different sources, such as maize (93.96%), wheat (85.24%), and cassava (79.03%). A semi-industrial scale reactor containing LOF-amy was prepared and showed the same yield of the laboratory-scale system. After five cycles of reuse, the LOF-amy reactor preserved over 80% of the initial amylase activity. Additionally, the LOF-amy was capable to operate as a kitchen grease trap component in a real situation during 30 days, preserving 30% of their initial amylase activity. PMID:23606948

  20. Immobilization of ?-Amylase onto Luffa operculata Fibers.

    PubMed

    Morais, Ricardo R; Pascoal, Aline M; Caramori, Samantha S; Lopes, Flavio M; Fernandes, Kátia F

    2013-01-01

    A commercial amylase (amy) was immobilized by adsorption onto Luffa operculata fibers (LOFs). The derivative LOF-amy presented capacity to hydrolyze starch continuously and repeatedly for over three weeks, preserving more than 80% of the initial activity. This system hydrolyzed more than 97% of starch during 5?min, at room temperature. LOF-amy was capable to hydrolyze starch from different sources, such as maize (93.96%), wheat (85.24%), and cassava (79.03%). A semi-industrial scale reactor containing LOF-amy was prepared and showed the same yield of the laboratory-scale system. After five cycles of reuse, the LOF-amy reactor preserved over 80% of the initial amylase activity. Additionally, the LOF-amy was capable to operate as a kitchen grease trap component in a real situation during 30 days, preserving 30% of their initial amylase activity. PMID:23606948

  1. Characterization of a new Bacillus stearothermophilus isolate: a highly thermostable ?-amylase-producing strain

    Microsoft Academic Search

    R. D. Wind; R. M. Buitelaar; G. Eggink; H. J. Huizing; L. Dijkhuizen

    1994-01-01

    A novel strain of Bacillus stearothermophilus was isolated from samples of a potato-processing industry. Compared to known a-amylases from other B. stearothermophilus strains, the isolate was found to produce a highly thermostable a-amylase. The half-time of inactivation of this a-amylase was 5.1 h at 80°C and 2.4 h at 90°C. The temperature optimum for activity of the a-amylase was 70°C;

  2. Effects of a new microbial ?-amylase inhibitor protein on Helicoverpa armigera larvae.

    PubMed

    Zeng, Fanrong; Wang, Xiaojing; Cui, Jinjie; Ma, Yan; Li, Qiannan

    2013-03-01

    A new microbial ?-amylase inhibitor gene was cloned and characterized. The encoded, recombinant, ?-amylase inhibitor protein was induced and expressed by isopropyl ?-d-1-thiogalactopyranoside (IPTG) in Escherichia coli M15 cells. The effects of the ?-amylase inhibitor protein on Helicoverpa armigera larvae were studied. Compared to the control, the weight of H. armigera larvae fed the diet with recombinant ?-amylase inhibitor protein added at a concentration of 20 ?g/g was reduced by 49.8%. The total soluble protein of H. armigera larvae fed the diet with the ?-amylase inhibitor protein added was also reduced by 36.8% compared to the control. The recombinant ?-amylase inhibitor protein showed inhibition activity against ?-amylase of H. armigera. These results suggested that this ?-amylase inhibitor protein may be a promising bioinsecticide candidate for controlling H. armigera. PMID:23432599

  3. Radionuclide salivary gland imaging

    SciTech Connect

    Mishkin, F.S.

    1981-10-01

    Salivary gland imaging with 99mTc as pertechnetate provides functional information concerning trapping and excretion of the parotid and submandibular glands. Anatomic information gained often adds little to clinical evaluation. On the other hand, functional information may detect subclinical involvement, which correlates well with biopsy of the minor labial salivary glands. Salivary gland abnormalities in systemic disease such as sarcoidosis, rheumatoid arthritis, lupus erythematosus, and other collagenvascular disorders may be detected before they result in the clinical manifestaions of Sjoegren's syndrome. Such glands, after initially demonstrating increased trapping in the acute phase, tend to have decreased trapping and failure to discharge pertechnetate in response to an appropriate physiologic stimulus. Increased uptake of gallium-67 citrate often accompanies these findings. Inflammatory parotitis can be suspected when increased perfusion is evident on radionuclide angiography with any agent. The ability of the salivary gland image to detect and categorize mass lesions, which result in focal areas of diminished activity such as tumors, cysts, and most other masses, is disappointing, while its ability to detect and categorize Warthin's tumor, which concentrates pertechnetate, is much more valuable, although not specific.

  4. Salt-dependent thermo-reversible ?-amylase: cloning and characterization of halophilic ?-amylase from moderately halophilic bacterium, Kocuria varians.

    PubMed

    Yamaguchi, Rui; Tokunaga, Hiroko; Ishibashi, Matsujiro; Arakawa, Tsutomu; Tokunaga, Masao

    2011-02-01

    A moderately halophilic bacterium, Kocuria varians, was found to produce active ?-amylase (K. varians ?-amylase (KVA)). We have observed at least six different forms of ?-amylase secreted by this bacterium into the culture medium. Characterization of these KVA forms and cloning of the corresponding gene revealed that KVA comprises pre-pro-precursor form of ?-amylase catalytic domain followed by the tandem repeats, which show high similarity to each other and to the starch binding domain (SBD) of other ?-amylases. The observed six forms were most likely derived by various processing of the protein product. Recombinant KVA protein was successfully expressed in Escherichia coli as a fusion protein and was purified with affinity chromatography after cleavage from fusion partner. The highly acidic amino acid composition of KVA and the highly negative electrostatic potential surface map of the modeled structure strongly suggested its halophilic nature. Indeed, KVA showed distinct salt- and time-dependent thermal reversibility: when ?-amylase was heat denatured at 85°C for 3 min in the presence of 2 M NaCl, the activity was recovered upon incubation on ice (50% recovery after 15 min incubation). Conversely, KVA denatured in 0.1 M NaCl was not refolded at all, even after prolonged incubation. KVA activity was inhibited by proteinaceous ?-amylase inhibitor from Streptomyces nitrosporeus, which had been implicated to inhibit only animal ?-amylases. KVA with putative SBD regions was found to digest raw starch. PMID:20871989

  5. Autonomous isolation, long-term culture and differentiation potential of adult salivary gland-derived stem/progenitor cells.

    PubMed

    Baek, Hyunjung; Noh, Yoo Hun; Lee, Joo Hee; Yeon, Soo-In; Jeong, Jaemin; Kwon, Heechung

    2014-09-01

    Salivary gland stem/progenitor cells belong to the endodermal lineage and may serve as good candidates to replace their dysfunctional counterparts. The objective of this study was to isolate large numbers of salivary gland tissue-derived stem cells (SGSCs) from adult rats in order to develop a clinically applicable method that does not involve sorting or stem cell induction by duct ligation. We analysed SGSCs isolated from normal rat salivary glands to determine whether they retained the major characteristics of stem cells, self-renewal and multipotency, especially with respect to the various endodermal cell types. SGSCs expressed high levels of integrin ?6?1 and c-kit, which are surface markers of SGSCs. In particular, the integrin ?6?1(+) /c-kit(+) salivary gland cells maintained the morphology, proliferation activity and multipotency of stem cells for up to 92 passages in 12 months. Furthermore, we analysed the capacity of SGSCs to differentiate into endoderm lineage cell types, such as acinar-like and insulin-secreting cells. When cultured on growth factor reduced matrigel, the morphology of progenitor cells changed to acinar-like structures and these cells expressed the acinar cell-specific marker, ?-amylase, and tight junction markers. Moreover, reverse transcription-polymerase chain reaction (RT-PCR) data showed increased expression of pancreatic cell markers, including insulin, Pdx1, pan polypeptide and neurogenin-3, when these cells formed pancreatic clusters in the presence of activin A, exendin-4 and retinoic acid. These data demonstrate that adult salivary stem/progenitor cells may serve as a potential source for cell therapy in salivary gland hypofunction and diabetes. PMID:22915381

  6. HPLC-DAD Analysis and In-Vitro Property of Polyphenols Extracts from (Solanum Aethiopium) Fruits on ? -Amylase, ? -Glucosidase and Angiotensin - 1- Converting Enzyme Activities

    PubMed Central

    Nwanna, E. E; Ibukun, E. O; Oboh, G.; Ademosun, A. O.; Boligon, A. A.; Athayde, M.

    2014-01-01

    AIM: Garden egg (Solanum aethiopium) is an edible fruits vegetable with  different species.This study investigated characterisation and the effect of the phenolics extracts from S. aethiopium species with enzymes linked with type -2-diabetes (?-amylase and ?-glucosidase) and hypertension [Angiotensin-1-converting enzyme (ACE)]. METHODS: Fresh samples of the 5 species of the garden egg namely, [Solanum gilo (PW), Solanum torvum (TWS), Solanum kumba (PGR), Solanum incanum (GSB), and Solanum indicum (WSB)] were oven-dried at 50°C and milled into flour. The aqueous extracts were prepared (1:50 w/v). The phenolic contents (total phenol and total flavonoid), vitamin C and 1,1-diphenyl–2-picrylhydrazyl (DPPH), the antioxidant activities of the extracts were evaluated. The ability of the extracts to inhibit diabetes enzymes in rat pancreas as well as the inhibition of angiotensin-1-converting (ACE) enzyme in lungs homogenates in vitro were investigated. Furthermore, the fruits polyphenols were identified and quantified using HPLC-DAD. RESULTS: The phenolic contents ranged from 2.70-3.76 mgGAE/g, while there were no significant (P>0.05) differences in their flavonoid content and ability to reduce Fe3+ to Fe2+. The vitamin C contents of the species ranged from 4.01-6.52 mg/ml. The extracts scavenged DPPH in a dose dependent manner with the IC50 values ranging from 3.23-4.20 mg/ml. Furthermore, the extracts showed strong inhibition of ?-glucosidase, mild inhibition of ?-amylase and strong inhibition of ACE activities. CONCLUSION: This study showed that the inhibition of the key enzymes relevant to type-2 diabetes and hypertension could be part of the mechanisms by which garden egg manage/prevent the degenerative conditions. PMID:25598760

  7. GA Enhanced a-Amylase Synthesis in Halved Grains of Barley (Hordeum vulgare): A Simple Laboratory Demonstration

    ERIC Educational Resources Information Center

    Freeland, P. W.

    1972-01-01

    A laboratory demonstration is suggested for the formation of a-amylase enzyme in halved grains of barley. Data presented in the article provide some information of the pattern of a- and b-amylase activity during germination. (PS)

  8. Adaptive significance of amylase polymorphism in Drosophila.

    E-print Network

    Paris-Sud XI, Université de

    Note Adaptive significance of amylase polymorphism in Drosophila. Analysis of the association between tissue-specific expression and specific activity in AmyS or AmyF genotypes of Drosophila in the midgut of adult Drosophila subobscura flies, homozygous for the Amys or AmyF allele, was analysed. The re

  9. Mass Spectrometric Analysis of Whole Secretome and Amylase-precipitated Secretome Proteins from Streptococcus gordonii

    PubMed Central

    Maddi, A; Haase, EM; Scannapieco, FA

    2014-01-01

    Oral biofilm (dental plaque) is formed by the initial adhesion of “pioneer species” to salivary proteins that form the dental pellicle on the tooth surface. One such pioneer species, Streptococcus gordonii, is known to bind salivary amylase through specific amylase-binding proteins such as amylase-binding protein A (AbpA). Recent studies have demonstrated that once bound, salivary amylase appears to modulate gene expression in S. gordonii. However, it is not known if this amylase-induced gene expression leads to secretion of proteins that play a role in plaque biofilm formation. In this study we examined the differences in secreted proteomes between S. gordonii KS1 (wild type) and AbpA-deficient (?AbpA) strains. We also examined the differentially precipitated secretome proteins following incubation with salivary amylase. The culture supernatants from KS1 and ?AbpA were analyzed by nano-LC/MS/MS to characterize the whole secreted proteomes of the KS1 and ?AbpA. A total of 107 proteins were identified in the KS1 and ?AbpA secretomes of which 72 proteins were predicted to have an N-terminal signal peptide for secretion. Five proteins were differentially expressed between the KS1 and ?AbpA secretomes; AbpA and sortase B were expressed exclusively by KS1, whereas Gdh, AdcA and GroEL were expressed only by ?AbpA. Incubation of culture supernatants from KS1 and ?AbpA with amylase (50 ?g/ml) at room temperature for 2 h resulted in the differential precipitation of secretome proteins. Hypothetical protein (SGO_0483), cation-transporting ATPase YfgQ (Aha1), isocitrate dehydrogenase (Icd), sortase A (SrtA), beta-N-acetylhexosaminidase (SGO_0405), peptide chain release factor 1(PrfA) and cardiolipin synthase (SGO_2037) were precipitated by amylase from the KS1 culture supernatant. Among the identified secreted proteins and amylase-precipitated proteins, transcriptional regulator LytR (SGO_0535) and cation-transporting ATPase YfgQ (Aha1) are potential signaling proteins. PMID:25605983

  10. High-resolution ?-amylase assay combined with high-performance liquid chromatography-solid-phase extraction-nuclear magnetic resonance spectroscopy for expedited identification of ?-amylase inhibitors: proof of concept and ?-amylase inhibitor in cinnamon.

    PubMed

    Okutan, Leyla; Kongstad, Kenneth T; Jäger, Anna K; Staerk, Dan

    2014-11-26

    Type 2 diabetes affects millions of people worldwide, and new improved drugs or functional foods containing selective ?-amylase inhibitors are needed for improved management of blood glucose. In this article the development of a microplate-based high-resolution ?-amylase inhibition assay with direct photometric measurement of ?-amylase activity is described. The inhibition assay is based on porcine pancreatic ?-amylase with 2-chloro-4-nitrophenyl-?-D-maltotriose as substrate, which this gives a stable, sensitive, and cheap inhibition assay as requested for high-resolution purposes. In combination with HPLC-HRMS-SPE-NMR, this provides an analytical platform that allows simultaneous chemical and biological profiling of ?-amylase inhibitors in plant extracts. Proof-of-concept with an artificial mixture of six compounds-of which three are known ?-amylase inhibitors-showed that the high-resolution ?-amylase inhibition profiles allowed detection of sub-microgram amounts of the ?-amylase inhibitors. Furthermore, the high-resolution ?-amylase inhibition assay/HPLC-HRMS-SPE-NMR platform allowed identification of cinnamaldehyde as the ?-amylase inhibitor in cinnamon (Cinnamomum verum Presl.). PMID:25368916

  11. The 53-kDa proteolytic product of precursor starch-hydrolyzing enzyme of Aspergillus niger has Taka-amylase-like activity.

    PubMed

    Ravi-Kumar, K; Venkatesh, K S; Umesh-Kumar, S

    2007-04-01

    The 53-kDa amylase secreted by Aspergillus niger due to proteolytic processing of the precursor starch-hydrolyzing enzyme was resistant to acarbose, a potent alpha-glucosidase inhibitor. The enzyme production was induced when A. niger was grown in starch medium containing the inhibitor. Antibodies against the precursor enzyme cross-reacted with the 54-kDa Taka-amylase protein of A. oryzae. It resembled Taka-amylase in most of its properties and also hydrolyzed starch to maltose of alpha-anomeric configuration. However, it did not degrade maltotriose formed during the reaction and was not inhibited by zinc ions. PMID:17123073

  12. Integrating Terminal Truncation and Oligopeptide Fusion for a Novel Protein Engineering Strategy To Improve Specific Activity and Catalytic Efficiency: Alkaline ?-Amylase as a Case Study

    PubMed Central

    Yang, Haiquan; Shin, Hyun-dong; Chen, Rachel R.; Li, Jianghua; Chen, Jian

    2013-01-01

    In this work, we integrated terminal truncation and N-terminal oligopeptide fusion as a novel protein engineering strategy to improve specific activity and catalytic efficiency of alkaline ?-amylase (AmyK) from Alkalimonas amylolytica. First, the C terminus or N terminus of AmyK was partially truncated, yielding 12 truncated mutants, and then an oligopeptide (AEAEAKAKAEAEAKAK) was fused at the N terminus of the truncated AmyK, yielding another 12 truncation-fusion mutants. The specific activities of the truncation-fusion mutants AmyK?C500-587::OP and AmyK?C492-587::OP were 25.5- and 18.5-fold that of AmyK, respectively. The kcat/Km was increased from 1.0 × 105 liters · mol?1 · s?1 for AmyK to 30.6 × and 23.2 × 105 liters · mol?1 · s?1 for AmyK?C500-587::OP and AmyK?C492-587::OP, respectively. Comparative analysis of structure models indicated that the higher flexibility around the active site may be the main reason for the improved catalytic efficiency. The proposed terminal truncation and oligopeptide fusion strategy may be effective to engineer other enzymes to improve specific activity and catalytic efficiency. PMID:23956385

  13. Integrating terminal truncation and oligopeptide fusion for a novel protein engineering strategy to improve specific activity and catalytic efficiency: alkaline ?-amylase as a case study.

    PubMed

    Yang, Haiquan; Liu, Long; Shin, Hyun-dong; Chen, Rachel R; Li, Jianghua; Du, Guocheng; Chen, Jian

    2013-10-01

    In this work, we integrated terminal truncation and N-terminal oligopeptide fusion as a novel protein engineering strategy to improve specific activity and catalytic efficiency of alkaline ?-amylase (AmyK) from Alkalimonas amylolytica. First, the C terminus or N terminus of AmyK was partially truncated, yielding 12 truncated mutants, and then an oligopeptide (AEAEAKAKAEAEAKAK) was fused at the N terminus of the truncated AmyK, yielding another 12 truncation-fusion mutants. The specific activities of the truncation-fusion mutants AmyK?C500-587::OP and AmyK?C492-587::OP were 25.5- and 18.5-fold that of AmyK, respectively. The kcat/Km was increased from 1.0 × 10(5) liters · mol(-1) · s(-1) for AmyK to 30.6 × and 23.2 × 10(5) liters · mol(-1) · s(-1) for AmyK?C500-587::OP and AmyK?C492-587::OP, respectively. Comparative analysis of structure models indicated that the higher flexibility around the active site may be the main reason for the improved catalytic efficiency. The proposed terminal truncation and oligopeptide fusion strategy may be effective to engineer other enzymes to improve specific activity and catalytic efficiency. PMID:23956385

  14. Alpha-amylase reactivity in relation to psychopathic traits in adults.

    PubMed

    Glenn, Andrea L; Remmel, Rheanna J; Raine, Adrian; Schug, Robert A; Gao, Yu; Granger, Douglas A

    2015-04-01

    Recent investigations of the psychobiology of stress in antisocial youth have benefited from a multi-system measurement model. The inclusion of salivary alpha-amylase (sAA), a surrogate marker of autonomic/sympathetic nervous system (ANS) activity, in addition to salivary cortisol, a biomarker of the hypothalamic-pituitary-adrenal (HPA) axis functioning, has helped define a more complete picture of individual differences and potential dysfunction in the stress response system of these individuals. To the authors' knowledge, no studies have examined sAA in relation to antisocial behavior in adults or in relation to psychopathic traits specifically. In the present study, we examined sAA, in addition to salivary cortisol, in a relatively large sample (n=158) of adult males (M age=36.81, range=22-67 years; 44% African-American, 34% Caucasian, 16% Hispanic) recruited from temporary employment agencies with varying levels of psychopathic traits. Males scoring highest in psychopathy were found to have attenuated sAA reactivity to social stress compared to those scoring lower in psychopathy. No differential relationships with the different factors of psychopathy were observed. In contrast to studies of antisocial youth, there were no interactions between sAA and cortisol levels in relation to psychopathy, but there was a significant interaction between pre-stressor levels of sAA and cortisol. Findings reveal potential regulatory deficits in the fast-acting, 'fight or flight', component of the stress response in adult males with psychopathic traits, as well as abnormalities in how this system may interact with the HPA axis. PMID:25662339

  15. The Rapalogue, CCI-779, Improves Salivary Gland Function following Radiation

    PubMed Central

    Morgan-Bathke, Maria; Harris, Zoey I.; Arnett, Deborah G.; Klein, Rob R.; Burd, Randy; Ann, David K.; Limesand, Kirsten H.

    2014-01-01

    The standard of care for head and neck cancer typically includes surgical resection of the tumor followed by targeted head and neck radiation. However depending on tumor location and stage, some cases may not require surgical resection while others may be treated with chemoradiation. Unfortunately, these radiation treatments cause chronic negative side effects for patients. These side effects are associated with damage to surrounding normal salivary gland tissue and include xerostomia, changes in taste and malnutrition. The underlying mechanisms of chronic radiation-induced salivary gland dysfunction are unknown, however, in rodent models persistently elevated proliferation is correlated with reduced stimulated salivary flow. The rapalogue, CCI-779, has been used in other cell systems to induce autophagy and reduce proliferation, therefore the aim of this study was to determine if CCI-779 could be utilized to ameliorate chronic radiation-induced salivary gland dysfunction. Four to six week old Atg5f/f; Aqp5-Cre, Atg5+/+; Aqp5-Cre and FVB mice were treated with targeted head and neck radiation. FVB mice were treated with CCI-779, chloroquine, or DMSO post-radiation. Stimulated salivary flow rates were determined and parotid and submandibular salivary gland tissues were collected for analyses. Mice with a defect in autophagy, via a conditional knockout of Atg5 in the salivary glands, display increased compensatory proliferation in the acinar cell compartment and hypertrophy at 24-72 hours following radiation. FVB mice treated with post-therapy CCI-779 have significant improvements in salivary gland physiology as determined by stimulated salivary flow rates, proliferation indices and amylase production and secretion. Consequently, post-radiation use of CCI-779 allows for improvement of salivary gland function and reestablishment of glandular homeostasis. As CCI-779 is already FDA approved for other uses, it could have a secondary use to alleviate the chronic side effects in head and neck cancer patients who have completed anti-tumor therapy. PMID:25437438

  16. Close relationship of a novel Flavobacteriaceae ?-amylase with archaeal ?-amylases and good potentials for industrial applications

    PubMed Central

    2014-01-01

    Background Bioethanol production from various starchy materials has received much attention in recent years. ?-Amylases are key enzymes in the bioconversion process of starchy biomass to biofuels, food or other products. The properties of thermostability, pH stability, and Ca-independency are important in the development of such fermentation process. Results A novel Flavobacteriaceae Sinomicrobium ?-amylase (FSA) was identified and characterized from genomic analysis of a novel Flavobacteriaceae species. It is closely related with archaeal ?-amylases in the GH13_7 subfamily, but is evolutionary distant with other bacterial ?-amylases. Based on the conserved sequence alignment and homology modeling, with minor variation, the Zn2+- and Ca2+-binding sites of FSA were predicated to be the same as those of the archaeal thermophilic ?-amylases. The recombinant ?-amylase was highly expressed and biochemically characterized. It showed optimum activity at pH 6.0, high enzyme stability at pH 6.0 to 11.0, but weak thermostability. A disulfide bond was introduced by site-directed mutagenesis in domain C and resulted in the apparent improvement of the enzyme activity at high temperature and broad pH range. Moreover, about 50% of the enzyme activity was detected under 100°C condition, whereas no activity was observed for the wild type enzyme. Its thermostability was also enhanced to some extent, with the half-life time increasing from 25 to 55 minutes at 50°C. In addition, after the introduction of the disulfide bond, the protein became a Ca-independent enzyme. Conclusions The improved stability of FSA suggested that the domain C contributes to the overall stability of the enzyme under extreme conditions. In addition, successfully directed modification and special evolutionary status of FSA imply its directional reconstruction potentials for bioethanol production, as well as for other industrial applications. PMID:24485248

  17. Responses of midgut amylases of Helicoverpa armigera to feeding on various host plants.

    PubMed

    Kotkar, Hemlata M; Sarate, Priya J; Tamhane, Vaijayanti A; Gupta, Vidya S; Giri, Ashok P

    2009-08-01

    Midgut digestive amylases and proteinases of Helicoverpa armigera, a polyphagous and devastating insect pest of economic importance have been studied. We also identified the potential of a sorghum amylase inhibitor against H. armigera midgut amylase. Amylase activities were detected in all the larval instars, pupae, moths and eggs; early instars had lower amylase levels which steadily increased up to the sixth larval instar. Qualitative and quantitative differences in midgut amylases of H. armigera upon feeding on natural and artificial diets were evident. Natural diets were categorized as one or more members of legumes, vegetables, flowers and cereals belonging to different plant families. Amylase activity and isoform patterns varied depending on host plant and/or artificial diet. Artificial diet-fed H. armigera larvae had comparatively high amylase activity and several unique amylase isoforms. Correlation of amylase and proteinase activities of H. armigera with the protein and carbohydrate content of various diets suggested that H. armigera regulates the levels of these digestive enzymes in response to macromolecular composition of the diet. These adjustments in the digestive enzymes of H. armigera may be to obtain better nourishment from the diet and avoid toxicity due to nutritional imbalance. H. armigera, a generalist feeder experiences a great degree of nutritional heterogeneity in its diet. An investigation of the differences in enzyme levels in response to macronutrient balance and imbalance highlight their importance in insect nutrition. PMID:19450602

  18. The Enzyme-Like Domain of Arabidopsis Nuclear ?-Amylases Is Critical for DNA Sequence Recognition and Transcriptional Activation[C][W][OPEN

    PubMed Central

    Soyk, Sebastian; Šimková, Klára; Zürcher, Evelyne; Luginbühl, Leonie; Brand, Luise H.; Vaughan, Cara K.; Wanke, Dierk; Zeeman, Samuel C.

    2014-01-01

    Plant BZR1-BAM transcription factors contain a ?-amylase (BAM)–like domain, characteristic of proteins involved in starch breakdown. The enzyme-derived domains appear to be noncatalytic, but they determine the function of the two Arabidopsis thaliana BZR1-BAM isoforms (BAM7 and BAM8) during transcriptional initiation. Removal or swapping of the BAM domains demonstrates that the BAM7 BAM domain restricts DNA binding and transcriptional activation, while the BAM8 BAM domain allows both activities. Furthermore, we demonstrate that BAM7 and BAM8 interact on the protein level and cooperate during transcriptional regulation. Site-directed mutagenesis of residues in the BAM domain of BAM8 shows that its function as a transcriptional activator is independent of catalysis but requires an intact substrate binding site, suggesting it may bind a ligand. Microarray experiments with plants overexpressing truncated versions lacking the BAM domain indicate that the pseudo-enzymatic domain increases selectivity for the preferred cis-regulatory element BBRE (BZR1-BAM Responsive Element). Side specificity toward the G-box may allow crosstalk to other signaling networks. This work highlights the importance of the enzyme-derived domain of BZR1-BAMs, supporting their potential role as metabolic sensors. PMID:24748042

  19. Neutrophil-Activating Protein Mediates Adhesion of Helicobacter pylori to Sulfated Carbohydrates on High-Molecular-Weight Salivary Mucin

    Microsoft Academic Search

    FERRY NAMAVAR; MARION SPARRIUS; ENNO C. I. VEERMAN; BEN J. APPELMELK

    1998-01-01

    The in vitro binding of surface-exposed material and outer membrane proteins of Helicobacter pylori to high-molecular-weight salivary mucin was studied. We identified a 16-kDa surface protein which adhered to high-molecular-weight salivary mucin. This protein binds specifically to sulfated oligosaccharide structures such as sulfo-Lewis a, sulfogalactose and sulfo-N-acetyl-glucosamine on mucin. Sequence analysis of the pro- tein proved that it was identical

  20. Substrate-inhibitor interactions in the kinetics of alpha-amylase inhibition by ragi alpha-amylase/trypsin inhibitor (RATI) and its various N-terminal fragments.

    PubMed

    Alam, N; Gourinath, S; Dey, S; Srinivasan, A; Singh, T P

    2001-04-10

    The ragi alpha-amylase/trypsin bifunctional inhibitor (RATI) from Indian finger millet, Ragi (Eleucine coracana Gaertneri), represents a new class of cereal inhibitor family. It exhibits a completely new motif of trypsin inhibitory site and is not found in any known trypsin inhibitor structures. The alpha-amylase inhibitory site resides at the N-terminal region. These two sites are independent of each other and the inhibitor forms a ternary (1:1:1) complex with trypsin and alpha-amylase. The trypsin inhibition follows a simple competitive inhibition obeying the canonical serine protease inhibitor mechanism. However, the alpha-amylase inhibition kinetics is a complex one if larger (> or =7 glucose units) substrate is used. While a complete inhibition of trypsin activity can be achieved, the inhibition of amylase is not complete even at very high molar concentration. We have isolated the N-terminal fragment (10 amino acids long) by CNBr hydrolysis of RATI. This fragment shows a simple competitive inhibition of alpha-amylase activity. We have also synthesized various peptides homologous to the N-terminal sequence of RATI. These peptides also show a normal competitive inhibition of alpha-amylase with varying potencies. It has also been shown that RATI binds to the larger substrates of alpha-amylase. In light of these observations, we have reexamined the binding of proteinaceous inhibitors to alpha-amylase and its implications on the mechanism and kinetics of inhibition. PMID:11284678

  1. Rescue of salivary gland function after stem cell transplantation in irradiated glands.

    PubMed

    Lombaert, Isabelle M A; Brunsting, Jeanette F; Wierenga, Pieter K; Faber, Hette; Stokman, Monique A; Kok, Tineke; Visser, Willy H; Kampinga, Harm H; de Haan, Gerald; Coppes, Robert P

    2008-01-01

    Head and neck cancer is the fifth most common malignancy and accounts for 3% of all new cancer cases each year. Despite relatively high survival rates, the quality of life of these patients is severely compromised because of radiation-induced impairment of salivary gland function and consequential xerostomia (dry mouth syndrome). In this study, a clinically applicable method for the restoration of radiation-impaired salivary gland function using salivary gland stem cell transplantation was developed. Salivary gland cells were isolated from murine submandibular glands and cultured in vitro as salispheres, which contained cells expressing the stem cell markers Sca-1, c-Kit and Musashi-1. In vitro, the cells differentiated into salivary gland duct cells and mucin and amylase producing acinar cells. Stem cell enrichment was performed by flow cytrometric selection using c-Kit as a marker. In vitro, the cells differentiated into amylase producing acinar cells. In vivo, intra-glandular transplantation of a small number of c-Kit(+) cells resulted in long-term restoration of salivary gland morphology and function. Moreover, donor-derived stem cells could be isolated from primary recipients, cultured as secondary spheres and after re-transplantation ameliorate radiation damage. Our approach is the first proof for the potential use of stem cell transplantation to functionally rescue salivary gland deficiency. PMID:18446241

  2. Lufaxin, a Novel Factor Xa Inhibitor from the Salivary Gland of the sand fly Lutzomyia longipalpis, Blocks PAR2 Activation and Inhibits Inflammation and Thrombosis in Vivo

    PubMed Central

    Collin, Nicolas; Assumpção, Teresa C. F.; Mizurini, Daniella M.; Gilmore, Dana; Dutra-Oliveira, Angélica; Kotsyfakis, Michalis; Sá-Nunes, Anderson; Teixeira, Clarissa; Ribeiro, José M. C.; Monteiro, Robson Q.; Valenzuela, Jesus G.; Francischetti, Ivo M. B.

    2012-01-01

    Objective Blood-sucking arthropods salivary glands (SGs) contain a remarkable diversity of antihemostatics. The aim of this study was to identify the unique salivary anticoagulant of the sand fly Lutzomyia longipalpis, which remained elusive for decades. Methods and Results Several L. longipalpis salivary proteins were expressed in HEK293 cells and screened for inhibition of blood coagulation. A novel 32.4-kDa molecule, named Lufaxin, was identified as a slow, tight, non-competitive, and reversible inhibitor of Factor Xa (FXa). Notably, Lufaxin primary sequence does not share similarity to any physiological or salivary inhibitors of coagulation reported to date. Lufaxin is specific for FXa and does not interact with FX, DEGR- FXa, or 15 other enzymes. In addition, Lufaxin blocks prothrombinase and increases both PT and aPTT. Surface plasmon resonance experiments revealed that FXa binds Lufaxin with a KD ~3 nM, and isothermal titration calorimetry determined a stoichiometry of 1:1. Lufaxin also prevents PAR2 activation by FXa in the MDA-MB-231 cell line and abrogates edema formation triggered by injection of FXa in the paw of mice. Moreover, Lufaxin prevents FeCl3-induced carotid artery thrombus formation and prolongs aPTT ex vivo, implying that it works as an anticoagulant in vivo. Finally, SG of sandflies was found to inhibit FXa and to interact with the enzyme. Conclusion Lufaxin belongs to a novel family of slow-tight FXa inhibitors, which display antithrombotic and antiinflamatory activities. It is a useful tool to understand FXa structural features and its role in pro-hemostatic and pro-inflammatory events. PMID:22796577

  3. What interactions drive the salivary mucosal pellicle formation?

    PubMed

    Gibbins, Hannah L; Yakubov, Gleb E; Proctor, Gordon B; Wilson, Stephen; Carpenter, Guy H

    2014-08-01

    The bound salivary pellicle is essential for protection of both the enamel and mucosa in the oral cavity. The enamel pellicle formation is well characterised, however the mucosal pellicle proteins have only recently been clarified and what drives their formation is still unclear. The aim of this study was to examine the salivary pellicle on particles with different surface properties (hydrophobic or hydrophilic with a positive or negative charge), to determine a suitable model to mimic the mucosal pellicle. A secondary aim was to use the model to test how transglutaminase may alter pellicle formation. Particles were incubated with resting whole mouth saliva, parotid saliva and submandibular/sublingual saliva. Following incubation and two PBS and water washes bound salivary proteins were eluted with two concentrations of SDS, which were later analysed using SDS-PAGE and Western blotting. Experiments were repeated with purified transglutaminase to determine how this epithelial-derived enzyme may alter the bound pellicle. Protein pellicles varied according to the starting salivary composition and the particle chemistry. Amylase, the single most abundant protein in saliva, did not bind to any particle indicating specific protein binding. Most proteins bound through hydrophobic interactions and a few according to their charges. The hydrophobic surface most closely matched the known salivary mucosal pellicle by containing mucins, cystatin and statherin but an absence of amylase and proline-rich proteins. This surface was further used to examine the effect of added transglutaminase. At the concentrations used only statherin showed any evidence of crosslinking with itself or another saliva protein. In conclusion, the formation of the salivary mucosal pellicle is probably mediated, at least in part, by hydrophobic interactions to the epithelial cell surface. PMID:24921197

  4. Human salivary gland stem cells ameliorate hyposalivation of radiation-damaged rat salivary glands.

    PubMed

    Jeong, Jaemin; Baek, Hyunjung; Kim, Yoon-Ju; Choi, Youngwook; Lee, Heekyung; Lee, Eunju; Kim, Eun Sook; Hah, Jeong Hun; Kwon, Tack-Kyun; Choi, Ik Joon; Kwon, Heechung

    2013-01-01

    Salivary function in mammals may be defective for various reasons, such as aging, Sjogren's syndrome or radiation therapy in head and neck cancer patients. Recently, tissue-specific stem cell therapy has attracted public attention as a next-generation therapeutic reagent. In the present study, we isolated tissue-specific stem cells from the human submandibular salivary gland (hSGSCs). To efficiently isolate and amplify hSGSCs in large amounts, we developed a culture system (lasting 4-5 weeks) without any selection. After five passages, we obtained adherent cells that expressed mesenchymal stem cell surface antigen markers, such as CD44, CD49f, CD90 and CD105, but not the hematopoietic stem cell markers, CD34 and CD45, and that were able to undergo adipogenic, osteogenic and chondrogenic differentiation. In addition, hSGSCs were differentiated into amylase-expressing cells by using a two-step differentiation method. Transplantation of hSGSCs to radiation-damaged rat salivary glands rescued hyposalivation and body weight loss, restored acinar and duct cell structure, and decreased the amount of apoptotic cells. These data suggest that the isolated hSGSCs, which may have characteristics of mesenchymal-like stem cells, could be used as a cell therapy agent for the damaged salivary gland. PMID:24232257

  5. Salivary parameters in infants aged 12 to 60 months with Down syndrome.

    PubMed

    Siqueira, Walter Luiz; Siqueira, Michelle Foigel; Mustacchi, Zan; de Oliveira, Elisabeth; Nicolau, José

    2007-01-01

    The purpose of this study was to measure certain components in whole saliva from children with Down syndrome aged 12 months to 60 months. Twenty children with Down syndrome were compared with 18 children without Down syndrome. Whole saliva was collected under slight suction and the salivary pH was measured with a portable pH meter soon after collection. Electrolyte concentrations were determined by inductively coupled argon plasma with atomic emission spectrometry. Sialic acid was determined by thiobarbituric acid assay. Amylase was assayed measuring the maltose produced by the breakdown of starch and peroxidase with ortho-dianisidine. No statistically significant differences were observed in sialic acid, calcium, phosphorus and magnesium concentrations between the group with Down syndrome and the control group. Protein and sodium concentration were higher in the group with Down syndrome compared to the control group. On the other hand, the flow rate, pH, amylase and peroxidase activities and potassium concentration were lower in those with Down syndrome compared to those children in the control group. PMID:17990480

  6. Alpha-amylases of the coffee berry borer (Hypothenemus hampei) and their inhibition by two plant amylase inhibitors.

    PubMed

    Valencia, A; Bustillo, A E; Ossa, G E; Chrispeels, M J

    2000-03-01

    The adult coffee berry borer (Hypothenemus hampei Ferrari [Coleoptera: Scolytidae]), a major insect pest of coffee, has two major digestive alpha-amylases that can be separated by isoelectric focusing. The alpha-amylase activity has a broad pH optimum between 4.0 and 7.0. Using pH indicators, the pH of the midgut was determined to be between 4.5 and 5.2. At pH 5.0, the coffee berry borer alpha-amylase activity is inhibited substantially (80%) by relatively low levels of the amylase inhibitor (alphaAI-1) from the common bean, Phaseolus vulgaris L., and much less so by the amylase inhibitor from Amaranthus. We used an in-gel zymogram assay to demonstrate that seed extracts can be screened to find suitable inhibitors of amylases. The prospect of using the genes that encode these inhibitors to make coffee resistant to the coffee berry borer via genetic engineering is discussed. PMID:10732988

  7. Cloning, nucleotide sequence, and enzymatic characterization of an alpha-amylase from the ruminal bacterium Butyrivibrio fibrisolvens H17c.

    PubMed Central

    Rumbak, E; Rawlings, D E; Lindsey, G G; Woods, D R

    1991-01-01

    A Butyrivibrio fibrisolvens amylase gene was cloned and expressed by using its own promoter on the recombinant plasmid pBAMY100 in Escherichia coli. The amylase gene consisted of an open reading frame of 2,931 bp encoding a protein of 976 amino acids with a calculated Mr of 106,964. In E. coli(pBAMY100), more than 86% of the active amylase was located in the periplasm, and TnphoA fusion experiments showed that the enzyme had a functional signal peptide. The B. fibrisolvens amylase is a calcium metalloenzyme, and three conserved putative calcium-binding residues were identified. The amylase showed high sequence homology with other alpha-amylases in the three highly conserved regions which constitute the active centers. These and other conserved regions were located in the N-terminal half, and no similarity with any other amylase was detected in the remainder of the protein. Deletion of approximately 40% of the C-terminal portion of the amylase did not result in loss of amylolytic activity. The B. fibrisolvens amylase was identified as an endo-alpha-amylase by hydrolysis of the Phadebas amylase substrate, hydrolysis of gamma-cyclodextrin to maltotriose, maltose, and glucose and the characteristic shape of the blue value and reducing sugar curves. Maltotriose was the major initial hydrolysis product from starch, although extended incubation resulted in its hydrolysis to maltose and glucose. Images PMID:2061294

  8. Optimization of Amylase Production from B. amyloliquefaciens (MTCC 1270) Using Solid State Fermentation

    PubMed Central

    Saha, Koel; Maity, Sujan; Roy, Sudeshna; Pathak, Rishija; Majumdar, Susmita

    2014-01-01

    Demand for microbial amylase production persists because of its immense importance in wide spectrum industries. The present work has been initiated with a goal of optimization of solid state fermentation condition for amylase using agroindustrial waste and microbial strain like B. amyloliquefaciens (MTCC 1270). In an aim to improve the productivity of amylase, fermentation has been carried out in the presence of calcium (Ca+2), Nitrate (NO3?), and chloride ions (Cl?) as well as in the presence of D-inositol and mannitol. Amylase needs calcium ion for the preservation of its structure, activity and stability that proves beneficial also for amylase production using solid state fermentation. The inclusion of ions and sugars in the SSF media is promising which can be explained by the protection offered by them against thermal decay of amylase at various incubation periods at 37°C. PMID:24949017

  9. Evaluation of alpha- amylase inhibition by Urtica dioica and Juglans regia extracts

    PubMed Central

    Rahimzadeh, Mahsa; Jahanshahi, Samaneh; Moein, Soheila; Moein, Mahmood Reza

    2014-01-01

    Objective(s): One strategy for the treatment of diabetes is inhibition of pancreatic ?- amylase. Plants contains different chemical constituents with potential for inhibition of ?-amylase and hence maybe used as therapeutic. Materials and Methods: Urtica dioica and Juglans regia Linn were tested for ?-amylase inhibition. Different concentrations of leaf aqueous extracts were incubated with enzyme substrate solution and the activity of enzyme was measured. For determination of the type of inhibition, Dixon plot was depicted. Acarbose was used as the standard inhibitor. Results: Both plant extracts showed time and concentration dependent inhibition of ?-amylase. 60% inhibition was seen with 2 mg/ml of U. dioica and 0.4 mg/ml of J. regia aqueous extract. Dixon plots revealed the type of ?-amylase inhibition by these two extracts as competitive inhibition. Conclusion: Determination of the type of ?-amylase inhibition by these plant extracts could provide by successful use of plant chemicals as drug targets. PMID:25140210

  10. Candida albicans flu1-mediated efflux of salivary histatin 5 reduces its cytosolic concentration and fungicidal activity.

    PubMed

    Li, Rui; Kumar, Rohitashw; Tati, Swetha; Puri, Sumant; Edgerton, Mira

    2013-04-01

    Histatin 5 (Hst 5) is a salivary human antimicrobial peptide that is toxic to the opportunistic yeast Candida albicans. Fungicidal activity of Hst 5 requires intracellular translocation and accumulation to a threshold concentration for it to disrupt cellular processes. Previously, we observed that total cytosolic levels of Hst 5 were gradually reduced from intact cells, suggesting that C. albicans possesses a transport mechanism for efflux of Hst 5. Since we identified C. albicans polyamine transporters responsible for Hst 5 uptake, we hypothesized that one or more polyamine efflux transporters may be involved in the efflux of Hst 5. C. albicans FLU1 and TPO2 were found to be the closest homologs of Saccharomyces cerevisiae TPO1, which encodes a major spermidine efflux transporter, indicating that the products of these two genes may be involved in efflux of Hst 5. We found that flu1?/? cells, but not tpo2?/? cells, had significant reductions in their rates of Hst 5 efflux and had significantly higher cytoplasmic Hst 5 and Hst 5 susceptibilities than did the wild type. We also found that flu1?/? cells had reduced biofilm formation compared to wild-type cells in the presence of Hst 5. Transcriptional levels of FLU1 were not altered over the course of treatment with Hst 5; therefore, Hst 5 is not likely to induce FLU1 gene overexpression as a potential mechanism of resistance. Thus, Flu1, but not Tpo2, mediates efflux of Hst 5 and is responsible for reduction of its toxicity in C. albicans. PMID:23380720

  11. Candida albicans Flu1-Mediated Efflux of Salivary Histatin 5 Reduces Its Cytosolic Concentration and Fungicidal Activity

    PubMed Central

    Li, Rui; Kumar, Rohitashw; Tati, Swetha; Puri, Sumant

    2013-01-01

    Histatin 5 (Hst 5) is a salivary human antimicrobial peptide that is toxic to the opportunistic yeast Candida albicans. Fungicidal activity of Hst 5 requires intracellular translocation and accumulation to a threshold concentration for it to disrupt cellular processes. Previously, we observed that total cytosolic levels of Hst 5 were gradually reduced from intact cells, suggesting that C. albicans possesses a transport mechanism for efflux of Hst 5. Since we identified C. albicans polyamine transporters responsible for Hst 5 uptake, we hypothesized that one or more polyamine efflux transporters may be involved in the efflux of Hst 5. C. albicans FLU1 and TPO2 were found to be the closest homologs of Saccharomyces cerevisiae TPO1, which encodes a major spermidine efflux transporter, indicating that the products of these two genes may be involved in efflux of Hst 5. We found that flu1?/? cells, but not tpo2?/? cells, had significant reductions in their rates of Hst 5 efflux and had significantly higher cytoplasmic Hst 5 and Hst 5 susceptibilities than did the wild type. We also found that flu1?/? cells had reduced biofilm formation compared to wild-type cells in the presence of Hst 5. Transcriptional levels of FLU1 were not altered over the course of treatment with Hst 5; therefore, Hst 5 is not likely to induce FLU1 gene overexpression as a potential mechanism of resistance. Thus, Flu1, but not Tpo2, mediates efflux of Hst 5 and is responsible for reduction of its toxicity in C. albicans. PMID:23380720

  12. Cloning and starch degradation profile of maltotriose-producing amylases from Streptomyces species.

    PubMed

    Kashiwagi, Norimasa; Miyake, Michiru; Hirose, Shuichi; Sota, Masahiro; Ogino, Chiaki; Kondo, Akihiko

    2014-11-01

    The end products from starch hydrolysis by amylases have important applications in various industries. Here, two amylases derived from two Streptomyces species that hydrolyze soluble starch from potato produced maltotriose as the primary maltooligosaccharide product. The genes, annotated as putative glycoside hydrolases, were cloned and expressed in Streptomyces lividans. These amylases displayed hydrolysis activity from pH 3 to 9.5 and were not affected by Ca(2+.) Optimal production of maltotriose was between 20 and 30 °C at pH 6.5. At the optimal temperature, both amylases produced maltotriose-rich end products rather than either maltose or maltotetraose. PMID:25048235

  13. Salivary cytokines as a minimally-invasive measure of immune functioning in young children: Correlates of individual differences and sensitivity to laboratory stress.

    PubMed

    Riis, Jenna L; Granger, Douglas A; DiPietro, Janet A; Bandeen-Roche, Karen; Johnson, Sara B

    2015-03-01

    There is growing interest in minimally-invasive measures of environmentally-responsive biological systems in developmental science. Contributing to that endeavor, this study explores the intercorrelations, correlates, and task-sensitivity of proinflammatory salivary cytokines in childhood. Saliva was sampled from 125 healthy five-year old children (49% male) across a series of cognitive and emotional challenge laboratory tasks. Samples were assayed for cytokines (IL-1?, IL-6, IL-8, TNF?), and markers of hypothalamic-pituitary-adrenal (HPA) and autonomic nervous system (ANS) activation (salivary cortisol and alpha-amylase [sAA]). Cytokines were positively intercorrelated and task-sensitivity varied. Except IL-8, cytokines were elevated in children with oral health issues and tobacco smoke exposure. Among boys, cytokines were positively related to sAA and negatively related to cortisol. The findings suggest that in healthy children, salivary cytokine levels reflect compartmentalized oral immune activity. Associations between ANS and HPA activity and cytokines in saliva may present opportunities for minimally-invasive methods to explore neuroendocrine-immune interactions during development. © 2015 Wiley Periodicals, Inc. Dev Psychobiol 57: 153-167, 2015. PMID:25604242

  14. Salivary gland disorders.

    PubMed

    Mandel, Louis

    2014-11-01

    Patients with salivary gland disease present with certain objective and/or subjective signs. An accurate diagnosis for these patients requires a range of techniques that includes the organized integration of information derived from their history, clinical examination, imaging, serology, and histopathology. This article highlights the signs and symptoms of the salivary gland disorders seen in the Salivary Gland Center, and emphasizes the methodology used to achieve a definitive diagnosis and therapy. PMID:25443682

  15. Regulation and genetic enhancement of beta-amylase production in Clostridium thermosulfurogenes.

    PubMed Central

    Hyun, H H; Zeikus, J G

    1985-01-01

    We studied the general mechanism for regulation of beta-amylase synthesis in Clostridium thermosulfurogenes. beta-Amylase was expressed at high levels only when the organism was grown on maltose or other carbohydrates containing maltose units. Three kinds of mutants altered in beta-amylase production were isolated by using nitrosoguanidine treatment, enrichment on 2-deoxyglucose, and selection of colonies with large clear zones on iodine-stained starch-glucose agar plates. beta-Amylase was produced only when maltose was added to cells growing on sucrose in wild-type and catabolite repression-resistant mutant strains, but the differential rate of enzyme synthesis in constitutive mutants was constant regardless of the presence of maltose. In carbon-limited chemostats of wild-type and catabolite repression-resistant mutant stains, beta-amylase was expressed on maltose but not on glucose or sucrose. beta-Amylase synthesis was immediately repressed by the addition of glucose. Therefore, we concluded that beta-amylase synthesis in C. thermosulfurogenes was inducible and subject to catabolite repression. The addition of cAMP did not eliminate the repressive effect of glucose. The mutants were generally characterized in terms of beta-amylase production, growth properties, fermentation product formation, and alterations in glucose isomerase and glucoamylase activities. A hyperproductive mutant produced eightfold more beta-amylase on starch medium than the wild type and more rapidly fermented starch to ethanol. PMID:2415505

  16. Capillary electrophoresis as a screening tool for alpha amylase inhibitors in plant extracts

    PubMed Central

    Hamdan, Imad I.; Afifi, Fatima U.

    2010-01-01

    Capillary electrophoresis (CE) method was developed for screening plant extract for potential alpha amylase (AA) inhibitory activity. The method was validated against a well established UV method. Overall, the proposed method was shown able to detect plants with significant alpha amylase inhibitory activity but not those with rather clinically insignificant activities. Fifty plant species were screened using both the proposed CE method and the UV method and seven plant species were found to possess significant AA inhibitory activities. Two plant species were proved to have alpha amylase inhibitory activity for the first time. PMID:24115900

  17. MALTOTRIOSE BRAKE: alpha-AMYLASE HYDROLYSIS PRODUCT MALTOTRIOSE REGULATES MALTASE-GLUCOAMYLASE ACTIVITY AND CONTROLS TOTAL RATES OF STARCH DIGESTION TO GLUCOSE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    BACKGROUND: Food starches provide 75% of meal-derived glucose required for normal brain energy supply. The digestion of cereals to glucose thus may be critically important in a weaning infant's diet. Human starch digestion is a multienzyme process involving six different enzymes. Salivary and pancre...

  18. Salivary Gland Cancer

    MedlinePLUS

    ... contains antibodies that can kill germs. Salivary gland cancer is a type of head and neck cancer. It is rare. It may not cause any ... pain in your face Doctors diagnose salivary gland cancer using a physical exam, imaging tests, and a ...

  19. Anopheles salivary gland proteomes from major malaria vectors

    PubMed Central

    2012-01-01

    Background Antibody responses against Anopheles salivary proteins can indicate individual exposure to bites of malaria vectors. The extent to which these salivary proteins are species-specific is not entirely resolved. Thus, a better knowledge of the diversity among salivary protein repertoires from various malaria vector species is necessary to select relevant genus-, subgenus- and/or species-specific salivary antigens. Such antigens could be used for quantitative (mosquito density) and qualitative (mosquito species) immunological evaluation of malaria vectors/host contact. In this study, salivary gland protein repertoires (sialomes) from several Anopheles species were compared using in silico analysis and proteomics. The antigenic diversity of salivary gland proteins among different Anopheles species was also examined. Results In silico analysis of secreted salivary gland protein sequences retrieved from an NCBInr database of six Anopheles species belonging to the Cellia subgenus (An. gambiae, An. arabiensis, An. stephensi and An. funestus) and Nyssorhynchus subgenus (An. albimanus and An. darlingi) displayed a higher degree of similarity compared to salivary proteins from closely related Anopheles species. Additionally, computational hierarchical clustering allowed identification of genus-, subgenus- and species-specific salivary proteins. Proteomic and immunoblot analyses performed on salivary gland extracts from four Anopheles species (An. gambiae, An. arabiensis, An. stephensi and An. albimanus) indicated that heterogeneity of the salivary proteome and antigenic proteins was lower among closely related anopheline species and increased with phylogenetic distance. Conclusion This is the first report on the diversity of the salivary protein repertoire among species from the Anopheles genus at the protein level. This work demonstrates that a molecular diversity is exhibited among salivary proteins from closely related species despite their common pharmacological activities. The involvement of these proteins as antigenic candidates for genus-, subgenus- or species-specific immunological evaluation of individual exposure to Anopheles bites is discussed. PMID:23148599

  20. Alpha-amylase from the Hyperthermophilic Archaeon Thermococcus thioreducens

    NASA Technical Reports Server (NTRS)

    Bernhardsdotter, E. C. M. J.; Pusey, M. L.; Ng, M. L.; Garriott, O. K.

    2003-01-01

    Extremophiles are microorganisms that thrive in, from an anthropocentric view, extreme environments such as hot springs. The ability of survival at extreme conditions has rendered enzymes from extremophiles to be of interest in industrial applications. One approach to producing these extremozymes entails the expression of the enzyme-encoding gene in a mesophilic host such as E.coli. This method has been employed in the effort to produce an alpha-amylase from a hyperthermophile (an organism that displays optimal growth above 80 C) isolated from a hydrothermal vent at the Rainbow vent site in the Atlantic Ocean. alpha-amylases catalyze the hydrolysis of starch to produce smaller sugars and constitute a class of industrial enzymes having approximately 25% of the enzyme market. One application for thermostable alpha-amylases is the starch liquefaction process in which starch is converted into fructose and glucose syrups. The a-amylase encoding gene from the hyperthermophile Thermococcus thioreducens was cloned and sequenced, revealing high similarity with other archaeal hyperthermophilic a-amylases. The gene encoding the mature protein was expressed in E.coli. Initial characterization of this enzyme has revealed an optimal amylolytic activity between 85-90 C and around pH 5.3-6.0.

  1. Effect of Cell Phone Use on Salivary Total Protein, Enzymes and Oxidative Stress Markers in Young Adults: A Pilot Study

    PubMed Central

    Joy, Jasmi; Sunitha, Venkatesh; Rai, Manoj P.; Rao, Suresh; Nambranathayil, Shafeeque; Baliga, Manjeshwar Shrinath

    2015-01-01

    Introduction: The present study aimed to assess the levels of salivary enzymes, protein and oxidant-antioxidant system in young college-going cell phone users. Materials and Methods: The cell users (students) were categorized in to two groups – less mobile users and high mobile users, based on the duration and frequency of cell use. Unstimulated whole saliva samples of the volunteers were analysed for amylase, lactate dehydrogenase (LDH), malondialdehdye (MDA) and glutathione (GSH). Results: High mobile users had significantly higher levels of amylase (p = 0.001), LDH (p = 0.002) and MDA (p = 0.002) in saliva, when compared to less mobile users. The marginal decrease in salivary total proteins, GSH and flow rate were statistically not significant (p >0.05). Conclusion: Significant changes in salivary enzymes and MDA suggest adverse effect of high use of cell phones on cell health.

  2. Salivary testosterone levels in preadolescent children

    PubMed Central

    Ostatníková, Daniela; Pastor, Karol; Putz, Zdenek; Dohnányiová, Monika; Mat'ašeje, Anna; Hampl, Richard

    2002-01-01

    Background Saliva reflects the plasma free fraction of testosterone which is biologically active, and available for uptake by tissues. Testosterone concentration in saliva, though differing slightly from the concentration of unbound testosterone in serum, is in good correlation with the latter, indicating that salivary testosterone provides a reliable method for determination of serum free testosterone. The study aimed to investigate salivary testosterone levels and their changes in preadolescent children and to study sexual dimorphism. Methods Testosterone levels were determined in 203 healthy preadolescent children (77 girls and 126 boys) from saliva samples by radioimmunoassay. Sampling was performed once a year with respect to circadian and seasonal fluctuations of testosterone. Data were statistically analyzed by Statgraphic software. Results Mean salivary testosterone concentrations (± SD) were 0.038 ± 0.012 nmol/L and 0.046 ± 0.026 nmol/L for girls and boys, with the medians 0.035 nmol/L and 0.041 nmol/L, respectively. Statistical analysis did not prove changes in salivary testosterone concentrations in the preadolescent period of life, with an exception of the insignificant fall at the age of 7 years, and an insignificant rise at the age of 9 years in girls. Conclusions Generally it can be concluded that salivary testosterone levels in our prepubertal subjects remained stable. There was no significant increase of salivary testosterone levels from the age of 6 until the age of 9 in both sexes. Sexual dimorphism in salivary testosterone levels was proved with significantly higher (p = 0.009) salivary testosterone levels in boys than in girls. PMID:12057024

  3. Treatment Options for Recurrent Salivary Gland Cancer

    MedlinePLUS

    ... for Clinical Trials NCI Publications Español Salivary Gland Cancer Treatment (PDQ®) Treatment Options for Recurrent Salivary Gland Cancer Treatment of recurrent salivary gland cancer may include the ...

  4. Purification, characterization, immunolocalization and structural analysis of the abundant cytoplasmic beta-amylase from Calystegia sepium (hedge bindweed) rhizomes.

    PubMed

    Van Damme, E J; Hu, J; Barre, A; Hause, B; Baggerman, G; Rougé, P; Peumans, W J

    2001-12-01

    An abundant catalytically active beta-amylase (EC 3.2.1.2) was isolated from resting rhizomes of hedge bindweed (Calystegia sepium). Biochemical analysis of the purified protein, molecular modeling, and cloning of the corresponding gene indicated that this enzyme resembles previously characterized plant beta-amylases with regard to its amino-acid sequence, molecular structure and catalytic activities. Immunolocalization demonstrated that the beta-amylase is exclusively located in the cytoplasm. It is suggested that the hedge bindweed rhizome beta-amylase is a cytoplasmic vegetative storage protein. PMID:11733023

  5. Production and Partial Purification of Alpha Amylase from Bacillus subtilis (MTCC 121) Using Solid State Fermentation.

    PubMed

    Raul, Dibyangana; Biswas, Tania; Mukhopadhyay, Suchita; Kumar Das, Shrayan; Gupta, Suvroma

    2014-01-01

    Amylase is an enzyme that catalyzes the breakdown of starch into sugars and plays a pivotal role in a variety of areas like use as digestives, for the production of ethanol and high fructose corn syrup, detergents, desiring of textiles, modified starches, hydrolysis of oil-field drilling fluids, and paper recycling. In the present work, solid state fermentation (SSF) for ? -amylase production has been used in lieu of submerged fermentation (SmF) due to its simple technique, low capital investment, lower levels of catabolite repression, and better product recovery. Bacillus subtilis has been well known as producer of alpha amylase and was tested using solid state fermentation for 48 hours at 37°C with wheat bran as substrate. Comparison between different fermentation hours demonstrated high yield of alpha amylase after 48 hours. This alpha amylase has optimum pH and temperature at 7.1 and 40°C, respectively. With the goal to purify alpha amylase, 30-70% (NH4)2SO4 cut concentrated the amylase activity threefold with respect to crude fermented extract. This was verified in quantitative DNS assay method as well as in zymogram gel profile. The exact molecular weight of the amylase is yet to be determined with the aid of other protein purification techniques. PMID:24672727

  6. Production and Partial Purification of Alpha Amylase from Bacillus subtilis (MTCC 121) Using Solid State Fermentation

    PubMed Central

    Raul, Dibyangana; Mukhopadhyay, Suchita; Kumar Das, Shrayan; Gupta, Suvroma

    2014-01-01

    Amylase is an enzyme that catalyzes the breakdown of starch into sugars and plays a pivotal role in a variety of areas like use as digestives, for the production of ethanol and high fructose corn syrup, detergents, desiring of textiles, modified starches, hydrolysis of oil-field drilling fluids, and paper recycling. In the present work, solid state fermentation (SSF) for ?-amylase production has been used in lieu of submerged fermentation (SmF) due to its simple technique, low capital investment, lower levels of catabolite repression, and better product recovery. Bacillus subtilis has been well known as producer of alpha amylase and was tested using solid state fermentation for 48 hours at 37°C with wheat bran as substrate. Comparison between different fermentation hours demonstrated high yield of alpha amylase after 48 hours. This alpha amylase has optimum pH and temperature at 7.1 and 40°C, respectively. With the goal to purify alpha amylase, 30–70% (NH4)2SO4 cut concentrated the amylase activity threefold with respect to crude fermented extract. This was verified in quantitative DNS assay method as well as in zymogram gel profile. The exact molecular weight of the amylase is yet to be determined with the aid of other protein purification techniques. PMID:24672727

  7. Dopamine-induced amylase secretion from guinea-pig submandibular gland.

    PubMed Central

    Bloom, G D; Carlsöö, B; Danielsson, A

    1975-01-01

    1. The effects of dopamine, 5-hydroxydopamine (5-OHDA) and noradrenaline on amylase secretion from the guinea-pig submandibular gland were investigated under in vitro conditions. 2. All three amines greatly enhanced amylase secretion. Blockade of dopamine beta-hydroxylase did not inhibit the response to dopamine. 3. Noradrenaline and dopamine stimulated amylase release from salivary glands of reserpine-treated animals, whereas 5-OHDA had no stimulatory effect on secretion in guinea-pigs pretreated with reserpine. 4. Haloperidol completely inhibited dopamine-induced enzyme discharge, but did not affect noradrenaline-initiated secretion. 5. Apomorphine caused a slight enzyme release by itself; it diminished the dopamine secretory effect, but did not modify that of noradrenaline. 6. Pimozide and fluspirilene attenuated the dopamine-induced enzyme discharge, but compared with haloperidol they were less effective. 7. It is concluded that dopamine exerts a secretagogic action different from that of noradrenaline. The possible presence of a specific dopamine receptor in salivary glands is discussed. PMID:169937

  8. Growth Defects Of Escherichia Coli Cells Which Contain The Gene Of An  Amylase from Bacillus coagulans on a Multicopy Plasmid

    Microsoft Academic Search

    KARINE WILLEMOT; PIERRE CORNELIS

    1983-01-01

    An a-amylase gene from Bacillus coagulans has previously been cloned in Escherichia coli and shown to direct the synthesis of an enzymically active protein of 60000 Dal (Cornelis et al., 1982). In one particular E. coli host, strain HBlO1, amylase was found to accumulate in the periplasmic space. To study the processing and the location of the amylase, plasmid pAMY2

  9. Purification and characterization of camel (Camelus dromedarius) milk amylase.

    PubMed

    El-Fakharany, Esmail M; Serour, Ehab A; Abdelrahman, Aref M; Haroun, Bakry M; Redwan, El-Rashdy M

    2009-01-01

    Skimmed camel milk contains 59,900 U/L amylase, which is 39,363 times less than serum and plasma amylase. Camel milk beta-amylase was purified as a 61 KDa band using DEAE-Sepharose and Sephadex G-100 and yielded 561 U/mg. The optimum working pH, Km and temperature were 7.0, 13.6 mg/Lstarch, 30-40 degrees C, respectively. The enzyme has been shown higher affinity toward amylose and soluble starch than glycogen, amylopectin, dextrin, or pullulan. Magnesium chloride, CaCl(2) and NaCl activated the amylase, while EDTA and EGTA decreased its activity. While its activity was increased in the presence of Triton X-100 and Triton X-114. Phenylmethanesulfonyl fluoride did not show any effect on enzyme activity. However, the enzyme activity was inhibited by urea, SDS, DTNB, iodoacetamide, N-ethylmalimide, aprotinin, and trypsin inhibitor. It worked on starch to yield a maltose. Scanning electron microscope images demonstrated a nano-degrading ability on starch granules from various sources (potato, corn, cassava, and rice). PMID:19291574

  10. Analysis of the Salivary Gland Transcriptome of Frankliniella occidentalis

    PubMed Central

    Stafford-Banks, Candice A.; Rotenberg, Dorith; Johnson, Brian R.; Whitfield, Anna E.; Ullman, Diane E.

    2014-01-01

    Saliva is known to play a crucial role in insect feeding behavior and virus transmission. Currently, little is known about the salivary glands and saliva of thrips, despite the fact that Frankliniella occidentalis (Pergande) (the western flower thrips) is a serious pest due to its destructive feeding, wide host range, and transmission of tospoviruses. As a first step towards characterizing thrips salivary gland functions, we sequenced the transcriptome of the primary salivary glands of F. occidentalis using short read sequencing (Illumina) technology. A de novo-assembled transcriptome revealed 31,392 high quality contigs with an average size of 605 bp. A total of 12,166 contigs had significant BLASTx or tBLASTx hits (E?1.0E?6) to known proteins, whereas a high percentage (61.24%) of contigs had no apparent protein or nucleotide hits. Comparison of the F. occidentalis salivary gland transcriptome (sialotranscriptome) against a published F. occidentalis full body transcriptome assembled from Roche-454 reads revealed several contigs with putative annotations associated with salivary gland functions. KEGG pathway analysis of the sialotranscriptome revealed that the majority (18 out of the top 20 predicted KEGG pathways) of the salivary gland contig sequences match proteins involved in metabolism. We identified several genes likely to be involved in detoxification and inhibition of plant defense responses including aldehyde dehydrogenase, metalloprotease, glucose oxidase, glucose dehydrogenase, and regucalcin. We also identified several genes that may play a role in the extra-oral digestion of plant structural tissues including ?-glucosidase and pectin lyase; and the extra-oral digestion of sugars, including ?-amylase, maltase, sucrase, and ?-glucosidase. This is the first analysis of a sialotranscriptome for any Thysanopteran species and it provides a foundational tool to further our understanding of how thrips interact with their plant hosts and the viruses they transmit. PMID:24736614

  11. THE EFFECT OF CHIROPRACTIC MANIPULATION ON SALIVARY CORTISOL LEVELS

    Microsoft Academic Search

    Tara L. Whelan; J. Donald Dishman; Jean Burke; Seymour Levine; Veronica Sciotti

    Background: The stress response in humans is a healthy response and is necessary for life. The effects of chiropractic manipulation (CM), if any, on stress are ill-defined. Cortisol has been used as an accurate measure of the stress response system in humans. Salivary cortisol is a noninvasive technique to accurately quantify biologically active cortisol. Objective: To determine whether basal salivary

  12. The effect of chiropractic manipulation on salivary cortisol levels

    Microsoft Academic Search

    Tara L. Whelan; J. Donald Dishman; Jean Burke; Seymour Levine; Veronica Sciotti

    2002-01-01

    Background: The stress response in humans is a healthy response and is necessary for life. The effects of chiropractic manipulation (CM), if any, on stress are ill-defined. Cortisol has been used as an accurate measure of the stress response system in humans. Salivary cortisol is a noninvasive technique to accurately quantify biologically active cortisol. Objective: To determine whether basal salivary

  13. Salivary gland tumors

    MedlinePLUS

    ... the salivary glands. In: Flint PW, Haughey BH, Lund VJ, et al., eds. Cummings Otolaryngology Head and ... NCCN Guidelines): Head and neck cancers. Version 2.2013. Available at: http://www.nccn.org/professionals/physician_ ...

  14. The Case for Primary Salivary Rhabdomyosarcoma

    PubMed Central

    Geltzeiler, Mathew; Li, Guangheng; Abraham, Jinu; Keller, Charles

    2015-01-01

    Rhabdomyosarcomas of the parotid and submandibular glands have the histological appearance of a skeletal muscle tumor yet can be found in tissue with no striated muscular elements. We examine the potential cell-of-origin for rhabdomyosarcoma and whether salivary tumors represent primary malignancy or metastasis. We have previously established genetically engineered mouse models of rhabdomyosarcoma. In these mice, rhabdomyosarcoma is only induced when a Pax3:Foxo1 fusion oncogene is activated with concurrent loss of p53 function (for alveolar rhabdomyosarcoma) or loss of p53 function alone (for embryonal rhabdomyosarcoma) using Cre-lox technology. These mutations are only activated under the control of promoters specific for selected cell lineages, previously thought to be myogenesis-restricted. RT-PCR and immunohistochemistry for lineage-specific promoter gene products reveal these promoters are active in wild-type mouse salivary gland. Given that mouse rhabdomyosarcoma frequently originates in the salivary glands and these myogenic-related promoters are normally expressed in salivary tissue, a high likelihood exists that the salivary gland contains a cell-of-origin of this muscle-related cancer. PMID:25883905

  15. H(1)-Receptor activation triggers the endogenous nitric oxide signalling system in the rat submandibular gland.

    PubMed Central

    Borda, Enri; Stranieri, Graciela; Sterin-Borda, Leonor

    2002-01-01

    BACKGROUND: Histamine is released from mast cells by immunologic and non-immunologic stimuli during salivary gland inflammation, regulating salivary secretion. The receptor-secretory mechanism has not been studied in detail. AIMS: The studies reported were directed toward elucidating signal transduction/second messenger pathways within the rat submandibular gland associated with 2-thiazolylethylamine (ThEA)-induced H(1)-receptor responses. MATERIALS AND METHODS: To assess the H(1) receptor subtype expression in the rat submandibular gland, a radioligand binding assay was performed. The study also included inositolphosphates and cyclic GMP accumulation, protein kinase C and nitric oxide synthase activities, and amylase release. RESULTS: The histamine H(1) receptor subtype is expressed on the rat submandibular gland with high-affinity binding sites. The ThEA effect was associated with activation of phosphoinositide-specific phospholipase C, translocation of protein kinase C, stimulation of nitric oxide synthase activity and increased production of cyclic GMP. ThEA stimulation of nitric oxide synthase and cyclic GMP was blunted by agents able to interfere with calcium movilization, while a protein kinase C inhibitor was able to stimulate ThEA action. On the other hand, ThEA stimulation evoked amylase release via the H1 receptor but was not followed by the L-arginine/nitric oxide pathway activation. CONCLUSIONS: These results suggest that, apart from the effect of ThEA on amylase release, it also appears to be a vasoactive chemical mediator that triggers vasodilatation, modulating the course of inflammation. PMID:12581497

  16. Salivary mucoepidermoid carcinoma revisited.

    PubMed

    Coca-Pelaz, Andrés; Rodrigo, Juan P; Triantafyllou, Asterios; Hunt, Jennifer L; Rinaldo, Alessandra; Strojan, Primož; Haigentz, Missak; Mendenhall, William M; Takes, Robert P; Vander Poorten, Vincent; Ferlito, Alfio

    2015-04-01

    Clinicopathological features, prognosis and therapeutic strategies for mucoepidermoid carcinoma originating in salivary and salivary-type glands of the head and neck are reviewed. We emphasise histopathological aspects, appraise the value of histochemistry, electron microscopy, immunohistochemistry and cytophotometry, and discuss histogenesis and characteristic gene translocations. We additionally consider possible diagnostic difficulties, problems related to histological grading and accuracy of existing literature, and areas of controversy or uncertainty which may benefit from further investigations. PMID:24771140

  17. Salivary Enzyme Polymorphisms (Set, Sgd and AMY1) in the Galician Population

    Microsoft Academic Search

    F. Boán; J. L. B. Caeiro

    1988-01-01

    The genetic polymorphism of three salivary enzymes (esterase, glucose-6-phosphate dehydrogenase and amylase) was studied in 580 autochthonous individuals from the Galician population (North-West Spain). The gene frequencies obtained were: SetF = 0.4036, Sets = 0.5964; Sgd1 – 0.7828, Sgd2 = 0.2172; AMY11 = 0.9319, AMY21 = 0.0495, AMY31 = 0.0186. Evidence of genetic intrapopulational heterogeneity was found for Set and

  18. Study on the correlations among disease activity index and salivary transforming growth factor-beta 1 and nitric oxide in ulcerative colitis patients.

    PubMed

    Rezaie, Ali; Khalaj, Sara; Shabihkhani, Maryam; Nikfar, Shekoufeh; Zamani, Mohammad J; Mohammadirad, Azadeh; Daryani, Naser E; Abdollahi, Mohammad

    2007-01-01

    Growth factors and nitric oxide (NO) play a major role in dysregulated immune response in ulcerative colitis (UC). Recent evidence has shown increased levels of transforming growth factor-beta(1) (TGF-beta(1)) in UC and suggested an anti-inflammatory effect for this factor. Based on our recent study, dysfunctional immunoregulation is present in saliva of UC patients, we hypothesized that salivary level of NO and TGF-beta(1) may differ by severity of UC and be useful to determine the activity of the disease. Thirty-seven UC patients and 15 healthy controls were enrolled and saliva samples were obtained. Truelove-Witts severity index and modified Truelove-Witts severity index were used to determine the severity of the disease. NO and TGF-beta(1) levels were detected in saliva of all patients and control subjects using enzyme-linked immunosorbent assay. A total of 21 patients had mild disease while 8 had moderate and 8 had severe colitis. Adjusted for baseline characteristics, the levels of NO and TGF-beta(1) in different groups were compared. Salivary NO and TGF-beta(1) levels were higher in UC patients comparing to controls (P < 0.00005 and P = 0.005, respectively). The levels of NO and TGF-beta(1) showed no significant differences among the severity groups (P = 0.46 and P = 0.23, respectively). NO levels linearly increased by age (Coeff = 1.5, r = 0.38, P = 0.02). Gender, extension of disease, and medical treatment did not affect NO and TGF-beta(1) levels. Although UC patients have abnormal amounts of NO and TGF-beta(1) in their saliva, their disease activity cannot be predicted by these factors, which may indicate a pathophysiologic role rather than being nonspecific inflammatory markers for TGF-beta(1) and NO. PMID:17404043

  19. Puffs and salivary gland function: The fine structure of the larval and prepupal salivary glands of Drosophila melanogaster

    Microsoft Academic Search

    Yvonne R. Carter; Michael Ashburner

    1972-01-01

    The salivary glands ofDrosophila melanogaster have been examined by electron microscopy for fine structural alterations occurring during larval and prepupal stages. The changes observed in the glands have been correlated with the puffing patterns of the polytene chromosomes at corresponding stages. In early third instar larvae, the lumen of the salivary gland appears empty, and no signs of secretory activity

  20. Discovery and characterization of pseudocyclic cystine-knot ?-amylase inhibitors with high resistance to heat and proteolytic degradation.

    PubMed

    Nguyen, Phuong Q T; Wang, Shujing; Kumar, Akshita; Yap, Li J; Luu, Thuy T; Lescar, Julien; Tam, James P

    2014-10-01

    Obesity and type 2 diabetes are chronic metabolic diseases, and those affected could benefit from the use of ?-amylase inhibitors to manage starch intake. The pseudocyclics, wrightides Wr-AI1 to Wr-AI3, isolated from an Apocynaceae plant show promise for further development as orally active ?-amylase inhibitors. These linear peptides retain the stability known for cystine-knot peptides in the presence of harsh treatment. They are resistant to heat treatment and endopeptidase and exopeptidase degradation, which is characteristic of cyclic cystine-knot peptides. Our NMR and crystallography analysis also showed that wrightides, which are currently the smallest proteinaceous ?-amylase inhibitors reported, contain the backbone-twisting cis-proline, which is preceded by a nonaromatic residue rather than a conventional aromatic residue. The modeled structure and a molecular dynamics study of Wr-AI1 in complex with yellow mealworm ?-amylase suggested that, despite having a similar structure and cystine-knot fold, the knottin-type ?-amylase inhibitors may bind to insect ?-amylase via a different set of interactions. Finally, we showed that the precursors of pseudocyclic cystine-knot ?-amylase inhibitors and their biosynthesis in plants follow a secretory protein synthesis pathway. Together, our findings provide insights for the use of the pseudocyclic ?-amylase inhibitors as useful leads for the development of orally active peptidyl bioactives, as well as an alternative scaffold for cyclic peptides for engineering metabolically stable human ?-amylase inhibitors. PMID:25040200

  1. MOLECULAR CLONING OF TRYPSIN-LIKE CDNAS AND COMPARISON OF PROTEINASE ACTIVITIES IN THE SALIVARY GLANDS AND GUT OF THE TARNISHED PLANT BUG LYGUS LINEOLARIS (HEMIPTERA: MIRIDAE)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Using specific proteinase inhibitors, we demonstrated that serine proteinases in the tarnished plant bug, Lygus lineolaris, are major proteinases in both salivary glands and gut tissues. Gut proteinases were less sensitive to inhibition than proteinases from the salivary glands. Up to 80% azocaseina...

  2. Deep metaproteomic analysis of human salivary supernatant.

    PubMed

    Jagtap, Pratik; McGowan, Thomas; Bandhakavi, Sricharan; Tu, Zheng Jin; Seymour, Sean; Griffin, Timothy J; Rudney, Joel D

    2012-04-01

    The human salivary proteome is extremely complex, including proteins from salivary glands, serum, and oral microbes. Much has been learned about the host component, but little is known about the microbial component. Here we report a metaproteomic analysis of salivary supernatant pooled from six healthy subjects. For deep interrogation of the salivary proteome, we combined protein dynamic range compression (DRC), multidimensional peptide fractionation, and high-mass accuracy MS/MS with a novel two-step peptide identification method using a database of human proteins plus those translated from oral microbe genomes. Peptides were identified from 124 microbial species as well as uncultured phylotypes such as TM7. Streptococcus, Rothia, Actinomyces, Prevotella, Neisseria, Veilonella, Lactobacillus, Selenomonas, Pseudomonas, Staphylococcus, and Campylobacter were abundant among the 65 genera from 12 phyla represented. Taxonomic diversity in our study was broadly consistent with metagenomic studies of saliva. Proteins mapped to 20 KEGG pathways, with carbohydrate metabolism, amino acid metabolism, energy metabolism, translation, membrane transport, and signal transduction most represented. The communities sampled appear to be actively engaged in glycolysis and protein synthesis. This first deep metaproteomic catalog from human salivary supernatant provides a baseline for future studies of shifts in microbial diversity and protein activities potentially associated with oral disease. PMID:22522805

  3. Deep metaproteomic analysis of human salivary supernatant

    PubMed Central

    Jagtap, Pratik; McGowan, Thomas; Bandhakavi, Sricharan; Tu, Zheng Jin; Seymour, Sean; Griffin, Timothy; Rudney, Joel

    2012-01-01

    The human salivary proteome is extremely complex, including proteins from salivary glands, serum, and oral microbes. Much has been learned about the host component, but little is known about the microbial component. Here we report a metaproteomic analysis of salivary supernatant pooled from 6 healthy subjects. For deep interrogation of the salivary proteome, we combined protein dynamic range compression, multidimensional peptide fractionation, and high mass accuracy MS/MS with a novel two-step peptide identification method using a database of human proteins plus those translated from oral microbe genomes. Peptides were identified from 124 microbial species as well as uncultured phylotypes such as TM7. Streptococcus, Rothia, Actinomyces, Prevotella, Neisseria, Veilonella, Lactobacillus, Selenomonas, Pseudomonas, Staphylococcus, and Campylobacter were abundant among the 65 genera from 12 phyla represented. Taxonomic diversity in our study was broadly consistent with metagenomic studies of saliva. Proteins mapped to twenty KEGG pathways, with Carbohydrate Metabolism, Amino Acid Metabolism, Energy Metabolism, Translation, Membrane Transport, and Signal Transduction most represented. The communities sampled appear to be actively engaged in glycolysis and protein synthesis. This first deep metaproteomic catalog from human salivary supernatant provides a baseline for future studies of shifts in microbial diversity and protein activities potentially associated with oral disease. PMID:22522805

  4. Factor Xa Activation of Factor V is of Paramount Importance in Initiating the Coagulation System: Lessons from a Tick Salivary Protein

    PubMed Central

    Schuijt, Tim J.; Bakhtiari, Kamran; Daffre, Sirlei; DePonte, Kathleen; Wielders, Simone J.H.; Marquart, J. Arnoud; Hovius, Joppe W.; van der Poll, Tom; Fikrig, Erol; Bunce, Matthew W.; Camire, Rodney M.; Nicolaes, Gerry A.F.; Meijers, Joost C.M.; van 't Veer, Cornelis

    2013-01-01

    Background Generation of active procoagulant cofactor FVa and its subsequent association with the enzyme FXa to form the prothrombinase complex is a pivotal initial event in blood coagulation and has been the subject of investigative effort, speculation and controversy. The current paradigm assumes that FV activation is initiated by limited proteolysis by traces of (meizo) thrombin. Methods and Results Recombinant tick salivary protein TIX-5 was produced and anticoagulant properties were studied using plasma, whole blood and purified systems. Here we report that TIX-5 specifically inhibits FXa-mediated FV activation involving the B-domain of FV and show that FXa activation of FV is pivotal for plasma and blood clotting. In line, tick feeding is impaired on TIX-5 immune rabbits displaying the in vivo importance of TIX-5. Conclusions Our data elucidate a unique molecular mechanism by which ticks inhibit the host's coagulation system. Based on our data we propose a revised blood coagulation scheme wherein direct FXa-mediated FV activation occurs in the initiation phase during which thrombin-mediated FV activation is restrained by fibrinogen and inhibitors. PMID:23817575

  5. Structural and biochemical features of acidic ?-amylase of Bacillus acidicola.

    PubMed

    Sharma, Archana; Satyanarayana, T

    2013-10-01

    The investigation is aimed at understanding structure-function aspect of ?-amylase of an acidophilic bacterium Bacillus acidicola (BAamy), which is Ca(2+)-independent and active at acidic pH of native starch, and thus, suits better in starch saccharification process. The CD spectroscopic data analysis revealed that the enzyme has 30% ?-helices, 14.2% ?-sheets, and 55.8% random coils at 60 °C and pH 4.0. Using Bacillus stearothermophilus ?-amylase (BStA) as the template, 3-D structure of rBAamy has been proposed. A complete loss in ?-amylase activity was recorded when the amino acid residues (D231, E261 and D328) were substituted that confirmed their role in catalysis. The CD studies indicated a decrease in the ?-helices content below and beyond the optimum pH and temperature that suggests a critical role of ?-helix in maintaining the structural conformation of the enzyme. Fluorescence-quenching by N-bromosuccinimide (NBS) suggested the role of tryptophan in maintaining structural integrity of ?-amylase and that by acrylamide indicated interaction by simple collision process. PMID:23954129

  6. A simple one pot purification of bacterial amylase from fermented broth based on affinity toward starch-functionalized magnetic nanoparticle.

    PubMed

    Paul, Tanima; Chatterjee, Saptarshi; Bandyopadhyay, Arghya; Chattopadhyay, Dwiptirtha; Basu, Semanti; Sarkar, Keka

    2015-08-18

    Surface-functionalized adsorbant particles in combination with magnetic separation techniques have received considerable attention in recent years. Selective manipulation on such magnetic nanoparticles permits separation with high affinity in the presence of other suspended solids. Amylase is used extensively in food and allied industries. Purification of amylase from bacterial sources is a matter of concern because most of the industrial need for amylase is met by microbial sources. Here we report a simple, cost-effective, one-pot purification technique for bacterial amylase directly from fermented broth of Bacillus megaterium utilizing starch-coated superparamagnetic iron oxide nanoparticles (SPION). SPION was prepared by co-precipitation method and then functionalized by starch coating. The synthesized nanoparticles were characterized by transmission electron microscopy (TEM), a superconducting quantum interference device (SQUID, zeta potential, and ultraviolet-visible (UV-vis) and Fourier-transform infrared (FTIR) spectroscopy. The starch-coated nanoparticles efficiently purified amylase from bacterial fermented broth with 93.22% recovery and 12.57-fold purification. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the molecular mass of the purified amylase was 67 kD, and native gel showed the retention of amylase activity even after purification. Optimum pH and temperature of the purified amylase were 7 and 50°C, respectively, and it was stable over a range of 20°C to 50°C. Hence, an improved one-pot bacterial amylase purification method was developed using starch-coated SPION. PMID:24840788

  7. Wheat grain preharvest sprouting and late maturity alpha-amylase.

    PubMed

    Mares, Daryl J; Mrva, Kolumbina

    2014-12-01

    Preharvest sprouting (PHS) and late maturity ?-amylase (LMA) are the two major causes of unacceptably high levels of ?-amylase in ripe wheat grain. High ?-amylase activity in harvested grain results in substantially lower prices for wheat growers and at least in the case of PHS, is associated with adverse effects on the quality of a range of end-products and loss of viability during storage. The high levels of ?-amylase are reflected in low falling number, the internationally accepted measure for grain receival and trade. Given the significant losses that can occur, elimination of these defects remains a major focus for wheat breeding programs in many parts of the world. In addition, the genetic, biochemical and molecular mechanisms involved in the control of PHS and LMA as well as the interactions with environmental factors have attracted a sustained research interest. PHS and LMA are independent, genetically controlled traits that are strongly influenced by the environment, where the effects of particular environmental factors vary substantially depending on the stage of grain development and ripening. This review is a summary and an assessment of results of recent research on these important grain quality defects. PMID:25257145

  8. Establishment of functional acinar-like cultures from human salivary glands.

    PubMed

    Jang, S I; Ong, H L; Gallo, A; Liu, X; Illei, G; Alevizos, I

    2015-02-01

    Disorders of human salivary glands resulting from therapeutic radiation treatment for head and neck cancers or from the autoimmune disease Sjögren syndrome (SS) frequently result in the reduction or complete loss of saliva secretion. Such irreversible dysfunction of the salivary glands is due to the impairment of acinar cells, the major glandular cells of protein, salt secretion, and fluid movement. Availability of primary epithelial cells from human salivary gland tissue is critical for studying the underlying mechanisms of these irreversible disorders. We applied 2 culture system techniques on human minor salivary gland epithelial cells (phmSG) and optimized the growth conditions to achieve the maintenance of phmSG in an acinar-like phenotype. These phmSG cells exhibited progenitor cell markers (keratin 5 and nanog) as well as acinar-specific markers-namely, ?-amylase, cystatin C, TMEM16A, and NKCC1. Importantly, with an increase of the calcium concentration in the growth medium, these phmSG cells were further promoted to acinar-like cells in vitro, as indicated by an increase in AQP5 expression. In addition, these phmSG cells also demonstrated functional calcium mobilization, formation of epithelial monolayer with high transepithelial electrical resistance (TER), and polarized secretion of ?-amylase secretion after ?-adrenergic receptor stimulation. Taken together, suitable growth conditions have been established to isolate and support culture of acinar-like cells from the human salivary gland. These primary epithelial cells can be useful for study of molecular mechanisms involved in regulating the function of acinar cells and in the loss of salivary gland function in patients. PMID:25416669

  9. Improved performance of ?-amylase immobilized on poly(glycidyl methacrylate-co-ethylenedimethacrylate) beads.

    PubMed

    He, Tian; Tian, Yong-Le; Qi, Liang; Zhang, Jing; Zhang, Zhi-Qi

    2014-04-01

    ?-Amylase was successfully immobilized onto poly(glycidyl methacrylate-co-ethylenedimethacrylate) (PGMA/EDMA) beads with high immobilization efficiency of 87.8%. PGMA/EDMA beads with a relatively uniform diameter of 2-3 ?m were prepared by single-step swelling polymerization. After amination with ethanediamine and activation with glutaraldehyde, PGMA/EDMA beads showed commendable ?-amylase immobilization capacity of 35.1 mg g(-1) carrier. Compared with free form, immobilized ?-amylase had increasement of 12.94 mg mL(-1) for Km and 0.124 mmol mL(-1) min(-1) for Vmax, improved acid resistance (the optimal pH from 7 to 5), presented better thermal stability by retaining 61% activity than 40% at 90 °C, and displayed high operational reusability by retaining 58% of its initial activity after nine uses. Moreover, less than 10% of the free enzyme and more than 80% of the immobilized enzyme retained activity after 180 min pre-incubation at 50 °C. The easy modification, high immobilization efficiency and good properties of immobilized ?-amylase in the present study indicate that PGMA/EDMA beads are suitable for ?-amylase immobilization. The enhancement of acid resistance and thermo stability is doubtless of benefit for the industrial use of ?-amylase. PMID:24518056

  10. Engineering ?-amylase levels in wheat grain suggests a highly sophisticated level of carbohydrate regulation during development.

    PubMed

    Whan, Alex; Dielen, Anne-Sophie; Mieog, Jos; Bowerman, Andrew F; Robinson, Hannah M; Byrne, Keren; Colgrave, Michelle; Larkin, Philip J; Howitt, Crispin A; Morell, Matthew K; Ral, Jean-Philippe

    2014-10-01

    Wheat starch degradation requires the synergistic action of different amylolytic enzymes. Our spatio-temporal study of wheat ?-amylases throughout grain development shows that AMY3 is the most abundant isoform compared with the other known ?-amylases. Endosperm-specific over-expression of AMY3 resulted in an increase of total ?-amylase activity in harvested grains. Unexpectedly, increased activity did not have a significant impact on starch content or composition but led to an increase of soluble carbohydrate (mainly sucrose) in dry grain. In AMY3 overexpression lines (A3OE), germination was slightly delayed and triacylglycerol (TAG) content was increased in the endosperm of mature grain. Despite increased AMY3 transcript and protein content throughout grain development, alterations of ?-amylase activity and starch granule degradation were not detected until grain maturation, suggesting a post-translational inhibition of ?-amylase activity in the endosperm during the starch filling period. These findings show unexpected effects of a high level of ?-amylase on grain development and composition, notably in carbon partitioning and TAG accumulation, and suggest the presence of a hitherto unknown regulatory pathway during grain filling. PMID:25053646

  11. Engineering ?-amylase levels in wheat grain suggests a highly sophisticated level of carbohydrate regulation during development

    PubMed Central

    Whan, Alex; Dielen, Anne-Sophie; Mieog, Jos; Bowerman, Andrew F.; Robinson, Hannah M.; Byrne, Keren; Colgrave, Michelle; Larkin, Philip J.; Howitt, Crispin A.; Morell, Matthew K.; Ral, Jean-Philippe

    2014-01-01

    Wheat starch degradation requires the synergistic action of different amylolytic enzymes. Our spatio-temporal study of wheat ?-amylases throughout grain development shows that AMY3 is the most abundant isoform compared with the other known ?-amylases. Endosperm-specific over-expression of AMY3 resulted in an increase of total ?-amylase activity in harvested grains. Unexpectedly, increased activity did not have a significant impact on starch content or composition but led to an increase of soluble carbohydrate (mainly sucrose) in dry grain. In AMY3 overexpression lines (A3OE), germination was slightly delayed and triacylglycerol (TAG) content was increased in the endosperm of mature grain. Despite increased AMY3 transcript and protein content throughout grain development, alterations of ?-amylase activity and starch granule degradation were not detected until grain maturation, suggesting a post-translational inhibition of ?-amylase activity in the endosperm during the starch filling period. These findings show unexpected effects of a high level of ?-amylase on grain development and composition, notably in carbon partitioning and TAG accumulation, and suggest the presence of a hitherto unknown regulatory pathway during grain filling. PMID:25053646

  12. Regulation of ?-amylase isoenzyme expression in Araucaria araucana by gibberellic and abscisic acids

    Microsoft Academic Search

    Elba Acevedo; Liliana Cardemil

    1997-01-01

    Expression of ?-amylase (EC 3.2.1.1) isoenzymes present in the embryos and mega-gametophytes of Araucaria araucana seedlings was studied under gibberellic acid (GA3) and abscisic acid (ABA) treatments and after 90 hr of imbibition in 20 mM of ?-chloroethyltrimethylammonium (CCC), a GA3 biosynthesis inhibitor. When CCC was used, ?-amylase activity in the treated embryos decreased to 28% with respect to the

  13. Characterization of ?-Amylase from Shoots and Cotyledons of Pea (Pisum sativum L.) Seedlings 1

    PubMed Central

    Beers, Eric P.; Duke, Stanley H.

    1990-01-01

    The most abundant ?-amylase (EC 3.2.1.1) in shoots and cotyledons from pea (Pisum sativum L.) seedlings was purified 6700-and 850-fold, respectively, utilizing affinity (amylose and cycloheptaamylose) and gel filtration chromatography and ultrafiltration. This ?-amylase contributed at least 79 and 15% of the total amylolytic activity in seedling cotyledons and shoots, respectively. The enzyme was identified as an ?-amylase by polarimetry, substrate specificity, and end product analyses. The purified ?-amylases from shoots and cotyledons appear identical. Both are 43.5 kilodalton monomers with pls of 4.5, broad pH activity optima from 5.5 to 6.5, and nearly identical substrate specificities. They produce identical one-dimensional peptide fingerprints following partial proteolysis in the presence of SDS. Calcium is required for activity and thermal stability of this amylase. The enzyme cannot attack maltodextrins with degrees of polymerization below that of maltotetraose, and hydrolysis of intact starch granules was detected only after prolonged incubation. It best utilizes soluble starch as substrate. Glucose and maltose are the major end products of the enzyme with amylose as substrate. This ?-amylase appears to be secreted, in that it is at least partially localized in the apoplast of shoots. The native enzyme exhibits a high degree of resistance to degradation by proteinase K, trypsin/chymostrypsin, thermolysin, and Staphylococcus aureus V8 protease. It does not appear to be a high-mannose-type glycoprotein. Common cell wall constituents (e.g. ?-glucan) are not substrates of the enzyme. A very low amount of this ?-amylase appears to be associated with chloroplasts; however, it is unclear whether this activity is contamination or ?-amylase which is integrally associated with the chloroplast. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 6 Figure 7 PMID:16667384

  14. Acid Diffusion into Rice Boluses is Influenced by Rice Type, Variety, and Presence of ?-Amylase.

    PubMed

    Mennah-Govela, Yamile A; Bornhorst, Gail M; Singh, R Paul

    2015-02-01

    Breakdown of rice during gastric digestion may be influenced by rice structure, presence of salivary ?-amylase, and hydrolysis by gastric acid. During mastication, saliva is mixed with rice, allowing ?-amylase to begin starch hydrolysis. This hydrolysis may continue in the gastric environment depending on the rate at which gastric acid penetrates into the rice bolus. The objective of this study was to determine the acid uptake into rice boluses with and without ?-amylase in saliva. Two types each of brown and white rice (medium and long grain), were formed into a cylindrical-shaped bolus. Each bolus was sealed on all sides except one to allow one-dimensional mass transfer, and incubated by immersion in simulated gastric juice at 37 °C under static conditions. Acidity of the boluses was measured by titration after 1 to 96 h of incubation. Effective diffusivity of the gastric juice through the bolus was estimated using MATLAB. Average acidity values ranged from 0.04 mg HCl/g dry matter (medium grain white rice, no incubation) to 10.01 mg HCl/g dry matter (long-grain brown rice, 72 h incubation). The rice type, presence of ?-amylase, and incubation time all significantly influenced rice bolus acidity (P < 0.001). Effective diffusivity of gastric juice into the bolus was greater in brown rice than in white rice. These results indicate that starch hydrolysis by ?-amylase may continue in the stomach before the gastric acid penetrates the rice bolus, and the rate of acid uptake will depend on the type of rice consumed. PMID:25559823

  15. Human cytomegalovirus and mucoepidermoid carcinoma of salivary glands: cell-specific localization of active viral and oncogenic signaling proteins is confirmatory of a causal relationship.

    PubMed

    Melnick, Michael; Sedghizadeh, Parish P; Allen, Carl M; Jaskoll, Tina

    2012-02-01

    Human cytomegalovirus (hCMV) infection is common. Although still controversial, there is growing evidence that active hCMV infection is associated with a variety of malignancies, including brain, breast, lung, colon, and prostate. Given that hCMV is frequently resident in salivary gland (SG) ductal epithelium, we hypothesized that hCMV would be important to the pathogenesis of SG mucoepidermoid carcinoma (MEC). This was initially supported by our finding that purified CMV induces malignant transformation in SG cells in an in vitro mouse model, and utilizes a pathogenic pathway previously reported for human MEC. Here we present the histologic and molecular characterizations of 39 human SG MECs selected randomly from a repository of cases spanning 2004-2011. Serial sections were obtained from formalin-fixed, paraffin embedded, tissue blocks from previous incisional or excisional biopsies. Immunohistochemical assays were performed for active hCMV proteins (IE1 and pp65) and the activated COX/AREG/EGFR/ERK signaling pathway. All four prospective causal criteria for viruses and cancer are fully satisfied: (1) protein markers for active hCMV are present in 97% of MECs; (2) markers of active hCMV are absent in non-neoplastic SG tissues; (3) hCMV-specific proteins (IE1, pp65) are in specific cell types and expression is positively correlated with severity; (4) hCMV correlates and colocalizes with an upregulation and activation of an established oncogenic signaling pathway (COX/AREG/EGFR/ERK). Thus, the evidential support reported here and previously in a mouse model is strongly confirmatory of a causal relationship between hCMV and SG mucoepidermoid carcinoma. To our knowledge, this is the first demonstration of hCMV's role in human oncogenesis that fully responds to all of Koch's Postulates as revised for viruses and cancer. In the absence of any contrary evidence, hCMV can reasonably be designated an "oncovirus." PMID:22101257

  16. Cloning and expression of alpha-amylase from the hyperthermophilic archaeon Pyrococcus woesei in the moderately halophilic bacterium Halomonas elongata.

    PubMed

    Frillingos, S; Linden, A; Niehaus, F; Vargas, C; Nieto, J J; Ventosa, A; Antranikian, G; Drainas, C

    2000-03-01

    An extracellular alpha-amylase gene from the hyperthermophilic archaeon Pyrococcus woesei has been cloned and sequenced. The 1.4-kb protein-coding sequence is identical to that of the corresponding alpha-amylase gene of the closely related species P. furiosus. By using a shuttle cloning vector for halophilic bacteria, the P. woesei alpha-amylase was expressed in the moderate halophile Halomonas elongata, under the control of a native H. elongata promoter. The hyperthermophilic amylase activity expressed in the halophilic host was recovered completely in the crude membrane fraction of cell homogenates, suggesting the formation of inclusion bodies or that the secretion machinery of H. elongata may fail to recognize and release the pyrococcal alpha-amylase to the extracellular medium. However, thermal stability, metal ion interactions, optimal temperature and pH values for the crude and purified recombinant alpha-amylase were comparable with those of the native pyrococcal enzyme. The P. woesei amylase activity expressed in H. elongata was consistently detected in the cells upon growth on a wide range of NaCl concentrations (0.7-2.5 mol l-1). To our knowledge, this is the first report on the expression of an archaeal gene (P. woesei alpha-amylase) in a moderate halophilic host which serves as a cell factory able to grow under extreme salt conditions and with very simple nutritional requirements. PMID:10747230

  17. Trastuzumab in Treating Patients With Metastatic or Recurrent Salivary Gland Cancer

    ClinicalTrials.gov

    2013-02-27

    High-grade Salivary Gland Mucoepidermoid Carcinoma; Recurrent Salivary Gland Cancer; Salivary Gland Acinic Cell Tumor; Salivary Gland Adenocarcinoma; Salivary Gland Poorly Differentiated Carcinoma; Stage IVA Salivary Gland Cancer; Stage IVB Salivary Gland Cancer; Stage IVC Salivary Gland Cancer

  18. Assessment of Salivary Hormones

    E-print Network

    Schultheiss, Oliver C.

    and behavior are differentiated. "Organizational effects" are lasting influ- ences that hormones exert on an individual's sex hormones (e.g., Graham & Desjardins, 1980; Roney, Lukaszewski, & Simmons, 2007); or when17 3 Assessment of Salivary Hormones Oliver C. Schultheiss Steven J. Stanton A Primer on Concepts

  19. Production of itaconic acid in Escherichia coli expressing recombinant ?-amylase using starch as substrate.

    PubMed

    Okamoto, Shusuke; Chin, Taejun; Nagata, Keisuke; Takahashi, Tetsuya; Ohara, Hitomi; Aso, Yuji

    2015-05-01

    Several studies on fermentative production of a vinyl monomer itaconic acid from hydrolyzed starch using Aspergillus terreus have been reported. Herein, we report itaconic acid production by Escherichia coli expressing recombinant ?-amylase, using soluble starch as its sole carbon source. To express ?-amylase in E. coli, we first constructed recombinant plasmids expressing ?-amylases by using cell surface display technology derived from two amylolytic bacteria, Bacillus amyloliquefaciens NBRC 15535(T) and Streptococcus bovis NRIC 1535. The recombinant ?-amylase from S. bovis (SBA) showed activity at 28°C, which is the optimal temperature for production of itaconic acid, while ?-amylase from B. amyloliquefaciens displayed no noticeable activity. E. coli cells expressing SBA produced 0.15 g/L itaconic acid after 69 h cultivation under pH-stat conditions, using 1% starch as the sole carbon source. In fact, E. coli cells expressing SBA had similar growth rates when grown in the presence of 1% glucose or starch, thereby highlighting the expression of an active ?-amylase that enabled utilization of starch to produce itaconic acid in E. coli. PMID:25468427

  20. Control of alpha-Amylase Development in Cotyledons during and following Germination of Mung Bean Seeds.

    PubMed

    Morohashi, Y; Katoh, H; Kaneko, Y; Matsushima, H

    1989-09-01

    Developmental patterns of alpha-amylase in Vigna radiata cotyledons during and following germination were quite different depending on the differences in the treatments of cotyledons during the imbibitional stage. When axis-detached cotyledons were imbibed in water with seed-coats attached, alpha-amylase activity did not increase and remained low. On the other hand, when the cotyledons were imbibed in water after seed-coat removal, the enzyme activity increased markedly. If the axis was attached to the cotyledons, alpha-amylase showed a marked development even under the former imbibition conditions. These changes in the enzyme activity were in parallel with those in the enzyme content, and the content, in turn, was dependent upon the availability of mRNA for alpha-amylase. We propose that the regulation of the development of alpha-amylase in cotyledons may involve some factor(s) inhibitory to accumulation of alpha-amylase mRNA, which is present in dry cotyledons and can be removed from cotyledons by leakage or by the presence of the axis. PMID:16667006

  1. Determination of the In Vitro and In Vivo Antimicrobial Activity on Salivary Streptococci and Lactobacilli and Chemical Characterisation of the Phenolic Content of a Plantago lanceolata Infusion.

    PubMed

    Ferrazzano, Gianmaria Fabrizio; Cantile, Tiziana; Roberto, Lia; Ingenito, Aniello; Catania, Maria Rosaria; Roscetto, Emanuela; Palumbo, Giuseppe; Zarrelli, Armando; Pollio, Antonino

    2015-01-01

    Introduction. Plant extracts may be suitable alternative treatments for caries. Aims. To investigate the in vitro and in vivo antimicrobial effects of Plantago lanceolata herbal tea (from flowers and leaves) on cariogenic bacteria and to identify the major constituents of P. lanceolata plant. Materials and Methods. The MIC and MBC against cariogenic bacteria were determined for P. lanceolata tea. Subsequently, a controlled random clinical study was conducted. Group A was instructed to rinse with a P. lanceolata mouth rinse, and Group B received a placebo mouth rinse for seven days. The salivary colonisation by streptococci and lactobacilli was investigated prior to treatment and on the fourth and seventh days. Finally, the P. lanceolata tea was analysed for its polyphenolic content, and major phenolics were identified. Results and Discussion. P. lanceolata teas demonstrate good in vitro antimicrobial activity. The in vivo test showed that Group A subjects presented a significant decrease in streptococci compared to Group B. The phytochemical analysis revealed that flavonoids, coumarins, lipids, cinnamic acids, lignans, and phenolic compounds are present in P. lanceolata infusions. Conclusions. P. lanceolata extract could represent a natural anticariogenic agent via an antimicrobial effect and might be useful as an ancillary measure to control the proliferation of cariogenic flora. PMID:25767805

  2. Determination of the In Vitro and In Vivo Antimicrobial Activity on Salivary Streptococci and Lactobacilli and Chemical Characterisation of the Phenolic Content of a Plantago lanceolata Infusion

    PubMed Central

    Roberto, Lia; Ingenito, Aniello; Roscetto, Emanuela

    2015-01-01

    Introduction. Plant extracts may be suitable alternative treatments for caries. Aims. To investigate the in vitro and in vivo antimicrobial effects of Plantago lanceolata herbal tea (from flowers and leaves) on cariogenic bacteria and to identify the major constituents of P. lanceolata plant. Materials and Methods. The MIC and MBC against cariogenic bacteria were determined for P. lanceolata tea. Subsequently, a controlled random clinical study was conducted. Group A was instructed to rinse with a P. lanceolata mouth rinse, and Group B received a placebo mouth rinse for seven days. The salivary colonisation by streptococci and lactobacilli was investigated prior to treatment and on the fourth and seventh days. Finally, the P. lanceolata tea was analysed for its polyphenolic content, and major phenolics were identified. Results and Discussion. P. lanceolata teas demonstrate good in vitro antimicrobial activity. The in vivo test showed that Group A subjects presented a significant decrease in streptococci compared to Group B. The phytochemical analysis revealed that flavonoids, coumarins, lipids, cinnamic acids, lignans, and phenolic compounds are present in P. lanceolata infusions. Conclusions. P. lanceolata extract could represent a natural anticariogenic agent via an antimicrobial effect and might be useful as an ancillary measure to control the proliferation of cariogenic flora. PMID:25767805

  3. Regulation of amylase, cellulase and chitinase secretion in the digestive tract of the two-spotted field cricket, Gryllus bimaculatus.

    PubMed

    Weidlich, Sandy; Müller, Sonja; Hoffmann, Klaus H; Woodring, Joseph

    2013-06-01

    The secretion of amylase and cellulase in Gryllus bimaculatus is determined by increased food intake, whereby shortly after molting food consumption increases. About half of the standing amylase concentration (activity) in the endothelial cells can be secreted within 30 min. The peak of amylase and cellulase secretion that occurs in the photophase is related to the feeding peak in the previous scotophase. The secretion of chitinase on the other hand is primarily controlled by the molting cycle. Only amylase secretion was affected by calcium in the incubation medium, suggesting an apocrine release mechanism. Refeeding experiments (after 5 days without food) suggest that the release of amylase in response to a nutrient in the lumen (glucose) is not due to simple stimulation of exocytosis, but rather a stimulation of synthesis. PMID:23585293

  4. Secreted expression of a hyperthermophilic ?-amylase gene from Thermococcus sp. HJ21 in Bacillus subtilis.

    PubMed

    Ying, Qi; Zhang, Chong; Guo, Fangfang; Wang, Shujun; Bie, Xiaomei; Lu, Fengxia; Lu, Zhaoxin

    2012-01-01

    The hyperthermophilic ?-amylase from Thermococcus sp. HJ21 possesses unique traits (Ca(2+)-independent thermostability and optimal temperature of 95°C) that make it a great potential candidate for use in the food industry. However, this Archaea isolated from a deep-sea thermal vent requires strict control of culture conditions and produces only small amounts of ?-amylase. To solve these problems, the ?-amylase gene was cloned and expressed in Bacillus subtilis, which is an ideal food-grade host for heterologous protein expression. To express high levels of this ?-amylase in B. subtilis, the promoters Pgrac, PxylA, P43, and Phag were used to construct four different expression vectors for testing. The vector containing the PxylA promoter was found to have the highest transcriptional activity and produce the highest amylase activity (19.6 U/ml). To test the secretion efficiency of signal peptides in B. subtilis, three signal peptides were cloned and fused to the ?-amylase gene (lacking its native signal peptide). The optimal signal peptide was SamyQ, with a secretion efficiency of approximately 90%. These results indicate that the promoter PxylA and signal peptide SamyQ tested in this study may be useful for the expression and secretion of archaeal proteins in B. subtilis. PMID:23486110

  5. [Salivary gland stem cells : Can they restore radiation-induced salivary gland dysfunction?].

    PubMed

    Rotter, N; Schwarz, S; Jakob, M; Brandau, S; Wollenberg, B; Lang, S

    2010-06-01

    Adult stem cells are actively investigated in the fields of regenerative medicine and tissue engineering, as they exhibit specific characteristics that make them promising candidates for cellular therapies. Depending on their tissue of origin these characteristics include long-term proliferation and the capacity to differentiate into various cell types. To date adult stem cells have been isolated from a multitude of tissues. Non-embryogenic adult tissues contain only small numbers of such stem cells and the derivation of such tissues can cause comorbidities. Therefore, there is ongoing interest in the identification and characterisation of novel cell sources for stem cell isolation and characterisation.Recently, salivary gland tissue has also been explored as a possible source of stem cells, first in animals and later in humans. Such salivary gland-derived stem cells might be useful in the treatment of radiation-induced salivary gland hypofunction, and possibly also in other diseases with loss of acinar cells, such as sequelae of radio iodine treatment or Sjögren's disease.In this paper we review the current status of salivary gland stem cell biology and application and discuss the possible role of stem cells in the development of novel therapies for salivary gland dysfunctions such as postradiogenic xerostomia. PMID:20464362

  6. Downregulation of p53 promotes in vitro perineural invasive activity of human salivary adenoid cystic carcinoma cells through epithelial-mesenchymal transition-like changes.

    PubMed

    Yang, Xiangming; Jing, Da; Liu, Lijun; Shen, Zhiyuan; Ju, Jun; Ma, Chao; Sun, Moyi

    2015-04-01

    Salivary adenoid cystic carcinoma (SACC) is a malignant tumor that is characterized by perineural invasion (PNI). p53 is an essential tumor-suppressor gene and p53 mutations play a critical role in tumor occurrence and progression (e.g., pancreatic, prostate and head and neck cancer). However, the regulatory role of the p53 gene in SACC and the PNI process remains unknown. In the present study, we employed RNA interference technique to downregulate p53 gene expression in SACC-83 cells to explore the role of p53 in the PNI process. Our results showed that the downregulation of the p53 gene induced significant 'epithelial-mesenchymal transition (EMT)-like changes' in SACC-83 cells, including decreased expression levels of epithelial markers (E-cadherin, EMA and CK5) and increased expression levels of mesenchymal markers (vimentin, N-cadherin and C-cadherin). The downregulation of p53 also caused a lower apoptotic index of Annexin V-FITC/PI and a lower number of SACC-83 cells in the second G0/G1 phase of the cell cycle. Furthermore, the downregulation of the p53 gene resulted in a significant increase in PNI activity in the SACC-83 cells. Thus, our findings revealed that downregulation of p53 promoted in vitro PNI activity through 'EMT-like changes' in SACC-83 cells. The present study suggests the essential regulatory role of p53 in the PNI activity of SACC cells, and implies that p53 may be a new target gene for the clinical treatment of SACC. PMID:25625376

  7. Arabidopsis thaliana AMY3 Is a Unique Redox-regulated Chloroplastic ?-Amylase*

    PubMed Central

    Seung, David; Thalmann, Matthias; Sparla, Francesca; Abou Hachem, Maher; Lee, Sang Kyu; Issakidis-Bourguet, Emmanuelle; Svensson, Birte; Zeeman, Samuel C.; Santelia, Diana

    2013-01-01

    ?-Amylases are glucan hydrolases that cleave ?-1,4-glucosidic bonds in starch. In vascular plants, ?-amylases can be classified into three subfamilies. Arabidopsis has one member of each subfamily. Among them, only AtAMY3 is localized in the chloroplast. We expressed and purified AtAMY3 from Escherichia coli and carried out a biochemical characterization of the protein to find factors that regulate its activity. Recombinant AtAMY3 was active toward both insoluble starch granules and soluble substrates, with a strong preference for ?-limit dextrin over amylopectin. Activity was shown to be dependent on a conserved aspartic acid residue (Asp666), identified as the catalytic nucleophile in other plant ?-amylases such as the barley AMY1. AtAMY3 released small linear and branched glucans from Arabidopsis starch granules, and the proportion of branched glucans increased after the predigestion of starch with a ?-amylase. Optimal rates of starch digestion in vitro was achieved when both AtAMY3 and ?-amylase activities were present, suggesting that the two enzymes work synergistically at the granule surface. We also found that AtAMY3 has unique properties among other characterized plant ?-amylases, with a pH optimum of 7.5–8, appropriate for activity in the chloroplast stroma. AtAMY3 is also redox-regulated, and the inactive oxidized form of AtAMY3 could be reactivated by reduced thioredoxins. Site-directed mutagenesis combined with mass spectrometry analysis showed that a disulfide bridge between Cys499 and Cys587 is central to this regulation. This work provides new insights into how ?-amylase activity may be regulated in the chloroplast. PMID:24089528

  8. Arabidopsis thaliana AMY3 is a unique redox-regulated chloroplastic ?-amylase.

    PubMed

    Seung, David; Thalmann, Matthias; Sparla, Francesca; Abou Hachem, Maher; Lee, Sang Kyu; Issakidis-Bourguet, Emmanuelle; Svensson, Birte; Zeeman, Samuel C; Santelia, Diana

    2013-11-22

    ?-Amylases are glucan hydrolases that cleave ?-1,4-glucosidic bonds in starch. In vascular plants, ?-amylases can be classified into three subfamilies. Arabidopsis has one member of each subfamily. Among them, only AtAMY3 is localized in the chloroplast. We expressed and purified AtAMY3 from Escherichia coli and carried out a biochemical characterization of the protein to find factors that regulate its activity. Recombinant AtAMY3 was active toward both insoluble starch granules and soluble substrates, with a strong preference for ?-limit dextrin over amylopectin. Activity was shown to be dependent on a conserved aspartic acid residue (Asp(666)), identified as the catalytic nucleophile in other plant ?-amylases such as the barley AMY1. AtAMY3 released small linear and branched glucans from Arabidopsis starch granules, and the proportion of branched glucans increased after the predigestion of starch with a ?-amylase. Optimal rates of starch digestion in vitro was achieved when both AtAMY3 and ?-amylase activities were present, suggesting that the two enzymes work synergistically at the granule surface. We also found that AtAMY3 has unique properties among other characterized plant ?-amylases, with a pH optimum of 7.5-8, appropriate for activity in the chloroplast stroma. AtAMY3 is also redox-regulated, and the inactive oxidized form of AtAMY3 could be reactivated by reduced thioredoxins. Site-directed mutagenesis combined with mass spectrometry analysis showed that a disulfide bridge between Cys(499) and Cys(587) is central to this regulation. This work provides new insights into how ?-amylase activity may be regulated in the chloroplast. PMID:24089528

  9. Management of Salivary Gland Cancer

    Microsoft Academic Search

    Laura D. Locati; Marco Guzzo; Patrizia Olmi; Lisa Licitra

    \\u000a Carcinomas of the salivary glands are uncommon representing only 2–6.5% of all head and neck cancer and less than 1% of all\\u000a cancers. About 85% of salivary gland tumors arise in the parotid glands and approximately 75% of these are benign while about\\u000a 75% of tumors arising from minor salivary glands are malignant. The latest WHO’s histological classification (2005) includes

  10. The receptor tyrosine kinase inhibitor vandetanib activates Akt and increases side population in a salivary gland tumor cell line (A253).

    PubMed

    Fujishiro, Yuka; Tonogi, Morio; Ochiai, Hiromi; Matsuzaka, Kenichi; Yamane, Gen-Yuki; Azuma, Toshifumi

    2012-07-01

    We and others have reported that cancer side population (SP) cells have self-renewal and multidrug resistance capabilities. These phenotypes are similar to those of cancer stem cells (CSCs), cancer stem-like cells and tumor-initiating cells (TICs). It has also been reported that upregulation of the epidermal growth factor receptor (EGFR) significantly increases the number of cancer SP cells, conversely, molecular targeting of EGFR tyrosine kinases using specific kinase inhibitors downregulates CSCs. Thus, we used flow cytometric analysis and cell sorting to examine cancer SP cells in the SCA9.cl-15, WR21 and A253 cell lines that originate from a salivary gland tumor (SGT). We successfully isolated cancer SP cells from all of these cell lines. SP cells were detected following treatment of these cell lines with the receptor tyrosine kinase inhibitors (RTKIs) lapatinib, erlotinib and vandetanib. Several studies reported that RTKIs mostly reduced the SP population in cancer cells. We did not observe any detectable morphological differences between SP cells and non-SP cells. We found that the EGF RTKI lapatinib decreased the number of cancer SP cells in all cell lines investigated; however, the EGF RTKI erlotinib did not cause significant differences in the frequency of cancer SP cells in these cell lines. Addition of vandetanib significantly increased the number of cancer SP cells and upregulated the phosphorylated Akt. As far as we know, this is the first report to show that one of the RTKIs, vandetanib, can activate Akt and increase the number of cancer SP cells. It has been reported that RTKIs could competitively inhibit ABC transporters and subsequently reduced the number of SP cells. However, our observation indicated that signaling changes induced by RTKIs could even activate Akt and induce the SP population. Investigation of the SP phenotype of SGTs is important for the establishment of optimal cancer therapy. PMID:22576692

  11. A study into salivary-based measurement of human stress subjected to Ellestad stress test protocol.

    PubMed

    Lee, Y K; Za'aba, A; Madzhi, N K; Ahmad, A

    2009-01-01

    Previous works on the effects of salivary alpha amylase in respond to various stressors report encouraging findings on it being a good indicator of stress. Ellestad protocol is a clinical procedure to screen for coronary artery disease by introducing exercise induced physical stress. If a salivary based biomarker profile in accordance to a stress test protocol could be established, the critical stress state which disable rational decision making could be ascertained in a standardized procedure. This technique would serve to aid human resource management in times of critical events such as rescue, firefighting or even military, that would potentially prevent unnecessary sacrifice of human lives. In this pilot study with five healthy volunteers performing the Ellestad protocol treadmill, a measurement profile with physiologic and salivary based biomarker is obtained. It is found that the alpha amylase levels or the changes in it as workload changes from resting-walking-running at ease-exhaustive running, is relatively more significant in reflecting the stress state than heart rate and blood pressure. Moreover, it is strongly associated with mood state with correlation coefficient of 0.8 and significance of 0.01. PMID:19964239

  12. C-terminal truncations of a thermostable Bacillus stearothermophilus alpha-amylase.

    PubMed

    Vihinen, M; Peltonen, T; Iitiä, A; Suominen, I; Mäntsälä, P

    1994-10-01

    A series of truncated proteins from a thermostable Bacillus stearothermophilus alpha-amylase was prepared to study the importance of the extension in the C-terminus compared with other liquefying Bacillus alpha-amylases. The mutations introducing new translation termination sites shortened the 515 amino acid residue-long wild type enzyme by 17, 32, 47, 73 or 93 residues. The longer the truncation, the lower the specific activity of the enzyme. Only the two longest mutant proteins were active: the specific activity of the 498 residue variant was 97% and protein 483 was 36% that of the parental enzyme. The Km values of starch hydrolysis changed from 1.09 for wild type enzyme to 0.35 and 0.21 for mutants 498 and 483, respectively, indicating altered substrate binding. The mutant enzymes had almost identical pH and temperature optima with the wild type amylase, but enhanced thermal stability and altered end product profile. The consequences of the truncation to the structure and function of the enzymes were explored with molecular modeling. The liquefying amylases seem to require approximately 480 residues to be active, whereas the C-terminal end of B.stearothermophilus amylase is required for increased activity. PMID:7855141

  13. Pre-therapeutic 124I PET(/CT) dosimetry confirms low average absorbed doses per administered 131I activity to the salivary glands in radioiodine therapy of differentiated thyroid cancer

    PubMed Central

    Hobbs, Robert F.; Stahl, Alexander; Knust, Jochen; Sgouros, George; Bockisch, Andreas

    2010-01-01

    Purpose Salivary gland impairment following high activity radioiodine therapy of differentiated thyroid cancer (DTC) is a severe side effect. Dosimetric calculations using planar gamma camera scintigraphy (GCS) with 131I and ultrasonography (US) provided evidence that the average organ dose per administered 131I activity (ODpA) is too low to account for observed radiation damages to the salivary glands. The objective of this work was to re-estimate the ODpA using 124I PET(/CT) as a more reliable approach than 131I GCS/US. Methods Ten DTC patients underwent a series of six (or seven) PET scans and one PET/CT scan after administration of ~23 MBq 124I-iodide. Volumes of interest (VOIs) drawn on the CT and serial PET images were used to determine the glandular volumes and the imaged 124I activities. To enable identical VOIs to be drawn on serial PET images, each PET was co-registered with the CT image. To correct for partial volume effect and for the artificial bias in the activity concentration due to cascading gamma coincidences occurring in 124I decay, the imaged activity was effectively corrected using isovolume recovery coefficients (RCs) based on recovery phantom measurements. A head-neck phantom, which contained 124I-filled spheres, was manufactured to validate the isovolume recovery correction method with a realistic patient-based phantom geometry and for a range of activity concentration regimes. The mean±standard deviation (range) ODpA projected for 131I was calculated using the absorbed dose fraction method. Results The ODpAs (in Gy/GBq) for the submandibular and parotid glands were 0.32±0.13 (0.18–0.55) and 0.31±0.10 (0.13–0.46), respectively. No significant differences (p>0.2) in the mean ODpA between 124I PET(/CT) and 131I GCS/US dosimetry was found. The validation experiment showed that the percentage deviations between RC-corrected and true activity concentrations were <10%. Conclusion 124I PET(/CT) dosimetry also corroborates the low ODpAs to the salivary glands. A voxel-based calculation taking into account the nonuniform activity distributions in the glands is necessary to possibly explain the radiation-induced salivary gland damage. PMID:20069293

  14. Isolation of a novel amylase and lipase-producing Pseudomonas luteola strain: study of amylase production conditions

    PubMed Central

    2014-01-01

    An amylase and lipase producing bacterium (strain C2) was enriched and isolated from soil regularly contaminated with olive washing wastewater in Sfax, Tunisia. Cell was aerobic, mesophilic, Gram-negative, motile, non-sporulating bacterium, capable of growing optimally at pH 7 and 30°C and tolerated maximally 10% (W/V) NaCl. The predominant fatty acids were found to be C18:1?7c (32.8%), C16:1?7c (27.3%) and C16:0 (23.1%). Phylogenetic analysis of the 16S rRNA gene revealed that this strain belonging to the genus Pseudomonas. Strain C2 was found to be closely related to Pseudomonas luteola with more than 99% of similarity. Amylase optimization extraction was carried out using Box Behnken Design (BBD). Its maximal activity was found when the pH and temperature ranged from 5.5 to 6.5 and from 33 to 37°C, respectively. Under these conditions, amylase activity was found to be about 9.48 U/ml. PMID:24405763

  15. Selection for low dormancy in annual ryegrass (Lolium rigidum) seeds results in high constitutive expression of a glucose-responsive ?-amylase isoform

    PubMed Central

    Goggin, Danica E.; Powles, Stephen B.

    2012-01-01

    Background and Aims ?-Amylase in grass caryopses (seeds) is usually expressed upon commencement of germination and is rarely seen in dry, mature seeds. A heat-stable ?-amylase activity was unexpectedly selected for expression in dry annual ryegrass (Lolium rigidum) seeds during targeted selection for low primary dormancy. The aim of this study was to characterize this constitutive activity biochemically and determine if its presence conferred insensitivity to the germination inhibitors abscisic acid and benzoxazolinone. Methods ?-Amylase activity in developing, mature and germinating seeds from the selected (low-dormancy) and a field-collected (dormant) population was characterized by native activity PAGE. The response of seed germination and ?-amylase activity to abscisic acid and benzoxazolinone was assessed. Using an alginate affinity matrix, ?-amylase was purified from dry and germinating seeds for analysis of its enzymatic properties. Key Results The constitutive ?-amylase activity appeared late during seed development and was mainly localized in the aleurone; in germinating seeds, this activity was responsive to both glucose and gibberellin. It migrated differently on native PAGE compared with the major activities in germinating seeds of the dormant population, but the enzymatic properties of ?-amylase purified from the low-dormancy and dormant seeds were largely indistinguishable. Seed imbibition on benzoxazolinone had little effect on the low-dormancy seeds but greatly inhibited germination and ?-amylase activity in the dormant population. Conclusions The constitutive ?-amylase activity in annual ryegrass seeds selected for low dormancy is electrophoretically different from that in germinating seeds and its presence confers insensitivity to benzoxazolinone. The concurrent selection of low dormancy and constitutive ?-amylase activity may help to enhance seedling establishment under competitive conditions. PMID:23002268

  16. The alpha-amylase gene amyH of the moderate halophile Halomonas meridiana: cloning and molecular characterization.

    PubMed

    Coronado, M J; Vargas, C; Mellado, E; Tegos, G; Drainas, C; Nieto, J J; Ventosa, A

    2000-04-01

    Two types of Tn1732-induced mutants defective in extracellular amylase activity were isolated from the moderate halophile Halomonas meridiana DSM 5425. Type I mutants displayed amylase activity in the periplasm, and were unable to use any of the carbon sources tested, including starch and its hydrolysis product maltose. The type II mutant was affected in the gene responsible for the synthesis of the extracellular alpha-amylase. This gene (amyH) was isolated by functional complementation of mutant II and sequenced. The deduced protein (AmyH) showed a high degree of homology to a proposed family of alpha-amylases consisting of enzymes from Alteromonas (Pseudoalteromonas) haloplanktis, Thermomonospora curvata, streptomycetes, insects and mammals. AmyH contained the four highly conserved regions in amylases, as well as a high content of acidic amino acids. The amyH gene was functional in the moderate halophile Halomonas elongata and, when cloned in a multicopy vector, in Escherichia coli. AmyH is believed to be the first extracellular-amylase-encoding gene isolated from a moderate halophile, a group of extremophiles of great biotechnological potential. In addition, H. meridiana and H. elongata were able to secrete the thermostable alpha-amylase from Bacillus licheniformis, indicating that members of the genus Halomonas are good candidates for use as cell factories to produce heterologous extracellular enzymes. PMID:10784044

  17. Gene cloning and characterization of a thermostable organic-tolerant ?-amylase from Bacillus subtilis DR8806.

    PubMed

    Emtenani, Shamsi; Asoodeh, Ahmad; Emtenani, Shirin

    2015-01-01

    The gene encoding an extracellular ?-amylase from Bacillus subtilis DR8806 was cloned into pET28a(+) vector and expressed in Escherichia coli BL21 (DE3). The recombinant enzyme with molecular mass of 76 kDa exhibited optimal activity at pH 5.0 and 70 °C with high stability in pH and temperature ranges of 4.0-9.0 and 45-75 °C. The enzyme showed a half-life of 125 min at 70 °C. The ?-amylase activity enhanced in the presence of Na(+), K(+), and Ca(2+) ions, while Zn(2+), Pb(2+), and Hg(2+) ions inhibited the activity. The recombinant ?-amylase exhibited high stability towards ioninc detergents sodium dodecyl sulfate (SDS) and cetyl trimethylammonium bromide (CTAB). Organic solvents in reaction media increased the ?-amylase activity. TLC analysis showed that maltoriose and maltose were the major end products of enzymatic starch hydrolysis. Presenting various properties of recombinant ?-amylase makes it well suited as a potential candidate for industrial usages. PMID:25168843

  18. Characterization of an organic solvent-tolerant ?-amylase from a halophilic isolate, Thalassobacillus sp. LY18.

    PubMed

    Li, Xin; Yu, Hui-Ying

    2012-09-01

    A halophilic isolate Thalassobacillus sp. LY18 producing extracellular amylase was isolated from the saline soil of Yuncheng Salt Lake, China. Production of the enzyme was synchronized with bacterial growth and reached a maximum level during the early stationary phase. The amylase was purified to homogeneity with a molecular mass of 31 kDa. Major products of soluble starch hydrolysis were maltose and maltotriose, indicating an ?-amylase activity. Optimal enzyme activity was found to be at 70°C, pH 9.0, and 10 % NaCl. The ?-amylase was highly stable over broad temperature (30-90°C), pH (6.0-12.0), and NaCl concentration (0-20 %) ranges, showing excellent thermostable, alkalistable, and halotolerant nature. The enzyme was stimulated by Ca(2+), but greatly inhibited by EDTA, indicating it was a metalloenzyme. Complete inhibition by diethyl pyrocarbonate and ?-mercaptoethanol revealed that histidine residue and disulfide bond were essential for enzyme catalysis. The surfactants tested had no significant effects on the amylase activity. Furthermore, it showed high activity and stability in the presence of water-insoluble organic solvents with log P (ow)???2.13. PMID:22581065

  19. Purification and Characterization of Midgut ?-Amylase in a Predatory Bug, Andralus spinidens

    PubMed Central

    Sorkhabi-Abdolmaleki, Sahar; Zibaee, Arash; Hoda, Hassan; Fazeli-Dinan, Mahmoud

    2014-01-01

    ?-Amylases are widespread enzymes that catalyze endohydrolysis of long ?-1,4-glucan chains such as starch and glycogen. The highest amylolytic activity was found in 5th instar nymphs and midgut of the predatory bug, Andrallus spinidens F. (Hemiptera: Pentatomidae). The ?-amylase was purified following a three-step procedure. The purified ?-amylase had a specific activity of 13.46 U/mg protein, recovery of 4.21, purification fold of 13.87, and molecular weight of 21.3 kDa. The enzyme had optimal pH and temperature of 7 and 45°C, respectively. Na+, Mn+, Mg2+, and Zn2+ significantly decreased activity of the purified ?-amylase, but some concentrations of K+, Ca2+, and Cu2+ had the opposite effect. EDTA, EGTA, and DTC significantly decreased enzymatic activity, showing the presence of metal ions in the catalytic site of the enzyme. Kinetic parameters of the purified ?-amylase showed a Km of 3.71% in starch and 4.96% for glycogen, suggesting that the enzyme had a higher affinity for starch. PMID:25373212

  20. Rhizopus microsporus var. rhizopodiformis: a thermotolerant fungus with potential for production of thermostable amylases.

    PubMed

    Peixoto, Simone C; Jorge, João A; Terenzi, Héctor F; Polizeli, Maria de Lourdes T M

    2003-12-01

    The effect of several nutritional and environmental parameters on growth and amylase production from Rhizopus microsporus var. rhizopodiformis was analysed. This fungus was isolated from soil of the Brazilian "cerrado" and produced high levels of amylolytic activity at 45 degrees C in liquid medium supplemented with starch, sugar cane bagasse, oat meal or cassava flour. Glucose in the culture medium drastically repressed the amylolytic activity. The products of hydrolysis were analysed by thin layer chromatography, and glucose was detected as the main component. The amylolytic activity hydrolysed several substrates, such as amylopectin, amylase, glycogen, pullulan, starch, and maltose. Glucose was always the main end product detected by high-pressure liquid chromatography analysis. These results indicated that the amylolytic activity studied is a glucoamylase, but there were also low levels of alpha-amylase. As compared to other fungi, R. microsporus var. rhizopodiformis can be considered an efficient producer of thermostable amylases, using raw residues of low cost as substrates. This information is of technological value, considering the importance of amylases for industrial hydrolysis. PMID:12920607

  1. Chronic activation of the epithelial immune system of the fruit fly's salivary glands has a negative effect on organismal growth and induces a peculiar set of target genes

    Microsoft Academic Search

    Ahmed Abdelsadik; Thomas Roeder

    2010-01-01

    BACKGROUND: Epithelial and especially mucosal immunity represents the first line of defence against the plethora of potential pathogens trying to invade via the gastrointestinal tract. The salivary glands of the fruit fly are an indispensable part of the gastrointestinal tract, but their contribution to the mucosal immunity has almost completely been neglected. Our major goal was to elucidate if the

  2. Stability and kinetics of a bifunctional amylase/trypsin inhibitor.

    PubMed

    Alagiri, S; Singh, T P

    1993-11-10

    The stability of the bifunctional amylase/trypsin inhibitor from ragi (Indian finger millet, Eleusine coracana) has been studied by methods of circular dichroism, UV absorption and intrinsic fluorescence. The inhibitor is stable in 8 M urea and 6 M guanidine-HCl. In 150 mM NaCl, thermal denaturation does not occur up to 90 degrees C. However, it is irreversibly denatured in 5 mM NaCl if heated over 73 degrees C. The acidic denaturation is reversible in both high and low salt conditions, but it shows different behavior below pH 1.65 under similar salt conditions. The helical content is about 2-4% in the pH range of 7-9 at which the inhibitor is active maximally. The NaCl concentration does not have a significant effect on the secondary structure elements. The beta-strand form does not show much variation under various conditions. Arg34-Leu35 is the reactive peptide bond in the trypsin-binding site. Trp and Tyr are involved in the binding with amylase. The bifunctional inhibitor represents the sum of individual inhibitors of trypsin and amylase. PMID:7692971

  3. Dasatinib in Treating Patients With Recurrent or Metastatic Malignant Salivary Gland Tumors

    ClinicalTrials.gov

    2015-04-02

    High-grade Salivary Gland Mucoepidermoid Carcinoma; Low-grade Salivary Gland Mucoepidermoid Carcinoma; Recurrent Salivary Gland Cancer; Salivary Gland Acinic Cell Tumor; Salivary Gland Adenocarcinoma; Salivary Gland Adenoid Cystic Carcinoma; Salivary Gland Anaplastic Carcinoma; Salivary Gland Malignant Mixed Cell Type Tumor; Salivary Gland Poorly Differentiated Carcinoma; Salivary Gland Squamous Cell Carcinoma; Stage IV Salivary Gland Cancer

  4. Salivary gland diseases in children

    PubMed Central

    Iro, Heinrich; Zenk, Johannes

    2014-01-01

    Salivary gland diseases in children are rare, apart from viral-induced diseases. Nevertheless, it is essential for the otolaryngologist to recognize these uncommon findings in children and adolescents and to diagnose and initiate the proper treatment. The present work provides an overview of the entire spectrum of congenital and acquired diseases of the salivary glands in childhood and adolescence. The current literature was reviewed and the results discussed and summarized. Besides congenital diseases of the salivary glands in children, the main etiologies of viral and bacterial infections, autoimmune diseases and tumors of the salivary glands were considered. In addition to the known facts, new developments in diagnostics, imaging and therapy, including sialendoscopy in obstructive diseases and chronic recurrent juvenile sialadenitis were taken into account. In addition, systemic causes of salivary gland swelling and the treatment of sialorrhoea were discussed. Although salivary gland diseases in children are usually included in the pathology of the adult, they differ in their incidence and some­times in their symptoms. Clinical diagnostics and especially the surgical treatment are influenced by a stringent indications and a less invasive strategy. Due to the rarity of tumors of the salivary glands in children, it is recommended to treat them in a specialized center with greater surgical experience. Altogether the knowledge of the differential diagnoses in salivary gland diseases in children is important for otolaryngologists, to indicate the proper therapeutic approach. PMID:25587366

  5. Isolation of a cDNA Clone for alpha-Amylase in Mung Bean Cotyledons : Analysis of alpha-Amylase mRNA Levels in Cotyledons during and following Germination of Mung Bean Seeds.

    PubMed

    Koizuka, N; Tanaka, Y; Morohashi, Y

    1990-11-01

    A cDNA was isolated that codes for alpha-amylase in mung bean (Vigna radiata) cotyledons, and the nucleotide sequence was determined. The deduced amino acid sequence (421 amino acid residues) is about 65% homologous with those of barley alpha-amylases. By comparing the deduced sequence with the sequence of the purified alpha-amylase, it was inferred that 23 N-terminal amino acids of a nascent polypeptide represent a signal peptide. Northern blot analysis showed that the levels of alpha-amylase mRNA are in parallel with the activities of alpha-amylase synthesis in cotyledons. Under the conditions where the solute leakage from cotyledons is accelerated during imbibition, a rapid increase in the amount of the alpha-amylase mRNA occurs. We postulate that a factor(s) which regulates in an inhibitory manner the alpha-amylase expression at the transcriptional level may be present in dry cotyledons and be removed by leakage. PMID:16667859

  6. Human Salivary Alpha-Amylase (EC.3.2.1.1) Activity and Periodic Acid and Schiff Reactive (PAS) Staining: A Useful Tool to Study Polysaccharides at an Undergraduate Level

    ERIC Educational Resources Information Center

    Fernandes, Ruben; Correia, Rossana; Fonte, Rosalia; Prudencio, Cristina

    2006-01-01

    Health science education is presently in discussion throughout Europe due to the Bologna Declaration. Teaching basic sciences such as biochemistry in a health sciences context, namely in allied heath education, can be a challenging task since the students of preclinical health sciences are not often convinced that basic sciences are clinically…

  7. Self-compassion training modulates alpha-amylase, heart rate variability, and subjective responses to social evaluative threat in women

    PubMed Central

    Arch, Joanna J.; Brown, Kirk Warren; Dean, Derek J.; Landy, Lauren N.; Brown, Kimberley; Laudenslager, Mark L.

    2014-01-01

    A growing body of research has revealed that social evaluative stressors trigger biological and psychological responses that in chronic forms have been linked to aging and disease. Recent research suggests that self-compassion may protect the self from typical defensive responses to evaluation. We investigated whether brief training in self-compassion moderated biopsychological responses to the Trier Social Stress Test (TSST) in women. Compared to attention (placebo) and no-training control conditions, brief self-compassion training diminished sympathetic (salivary alpha-amylase), cardiac parasympathetic, and subjective anxiety responses, though not HPA-axis (salivary cortisol) responses to the TSST. Self-compassion training also led to greater self-compassion under threat relative to the control groups. In that social stress pervades modern life, self-compassion represents a promising approach to diminishing its potentially negative psychological and biological effects. PMID:24636501

  8. Self-compassion training modulates alpha-amylase, heart rate variability, and subjective responses to social evaluative threat in women.

    PubMed

    Arch, Joanna J; Brown, Kirk Warren; Dean, Derek J; Landy, Lauren N; Brown, Kimberley D; Laudenslager, Mark L

    2014-04-01

    A growing body of research has revealed that social evaluative stressors trigger biological and psychological responses that in chronic forms have been linked to aging and disease. Recent research suggests that self-compassion may protect the self from typical defensive responses to evaluation. We investigated whether brief training in self-compassion moderated biopsychological responses to the Trier Social Stress Test (TSST) in women. Compared to attention (placebo) and no-training control conditions, brief self-compassion training diminished sympathetic (salivary alpha-amylase), cardiac parasympathetic, and subjective anxiety responses, though not HPA-axis (salivary cortisol) responses to the TSST. Self-compassion training also led to greater self-compassion under threat relative to the control groups. In that social stress pervades modern life, self-compassion represents a promising approach to diminishing its potentially negative psychological and biological effects. PMID:24636501

  9. Molecular basis for erythrocyte Le(a+b+) and salivary ABH partial-secretor phenotypes: expression of a FUT2 secretor allele with an A?T mutation at nucleotide 385 correlates with reduced ? (1,2) fucosyltransferase activity

    Microsoft Academic Search

    Stephen Henry; Rosella Mollicone; Pilar Fernandez; Bo Samuelsson; Rafael Oriol; Göran Larson

    1996-01-01

    TheSewA385T mutation of the FUT2 gene was found to correlate with both the erthrocyte Le(a+b+) and\\/or salivary ABH partial-secretor phenotypes of Polynesians. Constructs with FUT1 and FUT2 wild type genes, and the FUT2SewA385T,seG428A andseC571T mutated alleles, were cloned into pcDNAI, and expressed in COS-7 cells. COS-7 cells transfected with theSewA385T allele had weak, but detectable, ?(1,2)fucosyltransferase activity, with an acceptor

  10. Environmental and Genetic Contributors to Salivary Testosterone Levels in Infants

    PubMed Central

    Xia, Kai; Yu, Yang; Ahn, Mihye; Zhu, Hongtu; Zou, Fei; Gilmore, John H.; Knickmeyer, Rebecca C.

    2014-01-01

    Transient activation of the hypothalamic–pituitary–gonadal axis in early infancy plays an important role in male genital development and sexual differentiation of the brain, but factors contributing to individual variation in testosterone levels during this period are poorly understood. We measured salivary testosterone levels in 222 infants (119 males, 103 females, 108 singletons, 114 twins) between 2.70 and 4.80?months of age. We tested 16 major demographic and medical history variables for effects on inter-individual variation in salivary testosterone. Using the subset of twins, we estimated genetic and environmental contributions to salivary testosterone levels. Finally, we tested single nucleotide polymorphisms (SNPs) within ±5?kb of genes involved in testosterone synthesis, transport, signaling, and metabolism for associations with salivary testosterone using univariate tests and random forest (RF) analysis. We report an association between 5?min APGAR scores and salivary testosterone levels in males. Twin modeling indicated that individual variability in testosterone levels was primarily explained by environmental factors. Regarding genetic variation, univariate tests did not reveal any variants significantly associated with salivary testosterone after adjusting for false discovery rate. The top hit in males was rs10923844, an SNP of unknown function located downstream of HSD3B1 and HSD3B2. The top hits in females were two SNPs located upstream of ESR1 (rs3407085 and rs2295190). RF analysis, which reflects joint and conditional effects of multiple variants, indicated that genes involved in regulation of reproductive function, particularly LHCGR, are related to salivary testosterone levels in male infants, as are genes involved in cholesterol production, transport, and removal, while genes involved in estrogen signaling are related to salivary testosterone levels in female infants. PMID:25400620

  11. Characterization of two Acanthoscelides obtectus alpha-amylases and their inactivation by wheat inhibitors.

    PubMed

    Franco, Octávio L; Melo, Francislete R; Mendes, Paulo A; Paes, Norma S; Yokoyama, Massaru; Coutinho, Marise V; Bloch, Carlos; Grossi-de-Sá, Maria F

    2005-03-01

    Wheat alpha-amylase inhibitors represent an important tool in engineering crop plants against bean bruchids. Because Acanthoscelides obtectus is a devastating storage bean insect-pest, we attempted to purify and characterize its gut alpha-amylases, to study their interaction with active proteinaceous inhibitors. Two digestives alpha-amylases (AoA1 and AoA2) were purified from gut larvae, showing molecular masses of 30 and 45 kDa for each one, respectively. The stoichiometry interaction between these alpha-amylases with two wheat inhibitors (0.19 and 0.53) showed a binding complex of 1:1 enzyme:inhibitor. In vivo activities of these inhibitors against A. obtectus were also evaluated using a rich ammonium sulfate inhibitor fraction (F(20)(-)(40)) and purified inhibitors after reversed phase high-performance liquid chromatography columns. Incorporation of three different inhibitor concentrations (0.25, 0.5, and 1.0% w/w) into artificial seeds showed that addition of the purified 0.19 inhibitor at the highest concentration (1.0%) reduced the larval weight by 80%. Similar data were observed when 0.53 inhibitor was incorporated at 0.5%. When the concentration of purified 0.53 was enhanced to 1.0%, no larvae or adult emergence were observed. Our data suggest that these alpha-amylase inhibitors present great potential for use in Phaseolus genetic improvement programs. PMID:15740044

  12. Production and Biochemical Characterization of a High Maltotetraose (G4) Producing Amylase from Pseudomonas stutzeri AS22

    PubMed Central

    Maalej, Hana; Ben Ayed, Hanen; Ghorbel-Bellaaj, Olfa; Nasri, Moncef; Hmidet, Noomen

    2014-01-01

    Amylase production and biochemical characterization of the crude enzyme preparation from Pseudomonas stutzeri AS22 were evaluated. The highest ?-amylase production was achieved after 24 hours of incubation in a culture medium containing 10?g/L potato starch and 5?g/L yeast extract, with initial pH 8.0 at 30°C under continuous agitation at 200?rpm. The optimum temperature and pH for the crude ?-amylase activity were 60°C and 8.0, respectively. The effect of different salts was evaluated and it was found that both ?-amylase production and activity were Ca2+-dependent. The amylolytic preparation was found to catalyze exceptionally the formation of very high levels of maltotetraose from starch (98%, w/w) in the complete absence of glucose since the initial stages of starch hydrolysis (15?min) and hence would have a potential application in the manufacturing of maltotetraose syrups. PMID:24963472

  13. Bioactivity-Guided Separation of an ?-Amylase Inhibitor Flavonoid from Salvia virgata

    PubMed Central

    Nickavar, Bahman; Abolhasani, Leyla

    2013-01-01

    It is now believed that the inhibition of carbohydrate hydrolyzing enzymes (CHEs) in the digestive tract can significantly prolong the overall carbohydrate digestion time and decrease the postprandial hyperglycemia after a meal. Therefore, inhibitors of CHEs can be useful therapeutic approaches in the management of diabetes mellitus, especially in the type 2, and complications associated with the disease. In our previous study, the ethanol extract of the aerial parts of Salvia virgata showed an inhibitory effect on pancreatic ?-amylase in-vitro. Bioassay-guided fractionation of the extract using the ?-amylase inhibitory assay led to the isolation and identification of an active flavone compound, chrysoeriol. The compound concentration dependently inhibited the ?-amylase activity with an IC50 value of 1.27 (1.21-1.33) mM. PMID:24250572

  14. Host-mediated induction of alpha-amylases by larvae of the Mexican bean weevil Zabrotes subfasciatus (Coleoptera: Chrysomelidae: Bruchinae) is irreversible and observed from the initiation of the feeding period.

    PubMed

    Bifano, Thaís D; Samuels, Richard I; Alexandre, Daniel; Silva, Carlos P

    2010-08-01

    Larvae of Zabrotes subfasciatus secrete alpha-amylases that are insensitive to the alpha-amylase inhibitor found in seeds of Phaseolus vulgaris. By analyzing amylase activities during larval development on P. vulgaris, we detected activity of the constitutive amylase and the two inducible amylase isoforms at all stages. When larvae were transferred from the non alpha-amylase inhibitor containing seeds of Vigna unguiculata to P. vulgaris, the inducible alpha-amylases were expressed at the same level as in control larvae fed on P. vulgaris. Interestingly, when larvae were transferred from seeds of P. vulgaris to those of V. unguiculata, inducible alpha-amylases continued to be expressed at a level similar to that found in control larvae fed P. vulgaris continuously. When 10-day-old larvae were removed from seeds of V. unguiculata and transferred into capsules containing flour of P. vulgaris cotyledons, and thus maintained until completing 17 days (age when the larvae stopped feeding), we could detect higher activity of the inducible alpha-amylases. However, when larvae of the same age were transferred from P. vulgaris into capsules containing flour of V. unguiculata, the inducible alpha-amylases remained up-regulated. These results suggest that the larvae of Z. subfasciatus have the ability to induce insensitive amylases early in their development. A short period of feeding on P. vulgaris cotyledon flour was sufficient to irreversibly induce the inducible alpha-amylase isoforms. Incubations of brush border membrane vesicles with the alpha-amylase inhibitor 1 from P. vulgaris suggest that the inhibitor is recognized by putative receptors found in the midgut microvillar membranes. PMID:20645418

  15. Seasonal variation in amylase, invertase, cellulase activity and carbon dioxide evolution in a tropical protected grassland of Orissa, India, sprayed with carbaryl insecticide.

    PubMed

    Mishra, P C; Pradhan, S C

    1987-01-01

    A distinct seasonal variation in the enzyme activities and carbon dioxide evolution in soil, with a maximum in summer, was observed in soil treated with carbaryl and in control soil. There was no significant difference in the rate of enzyme activity between 0 and 10 cm and 10 and 20 cm depth of soil. Carbaryl insecticide applied at a normal agricultural dose did not have any inhibitory effect on soil enzyme activity, or on the rate of CO(2) evolution. However, the cellulase activity was greater in the surface soil of the control plot than in the treated plot. PMID:15092792

  16. Purification and characterization of a truncated Bacillus subtilis ?-amylase produced by Escherichia coli

    Microsoft Academic Search

    J. L. Marco; L. A. Bataus; F. F. Valência; C. J. Ulhoa; S. Astolfi-Filho; C. R. Felix

    1996-01-01

    ABacillus subtilis amylase gene was inserted into a plasmid which transferred toEscherichia coli. During cloning, a 3' region encoding 171 carboxyterminal amino acids was replaced by a nucleotide sequence that encoded 33 amino acid residues not present in the indigenous protein. The transformed cells produced substantial amylolytic activity. The active protein was purified to apparent homogeneity. Its molecular mass (48

  17. Effect of cadmium on germination, amylases and rate of respiration of germinating pea seeds

    Microsoft Academic Search

    L. K. Chugh; S. K. Sawhney

    1996-01-01

    Growth of embryonic axis of germinating pea seeds (Pisum sativum cv. Bonneville) was significantly inhibited by as low as 0.25 mM cadmium and the elongation of the radicle was affected more severely than that of the plumule. Total amylolytic activity, as well as activities of ?- and ?-amylases, diminished progressively with increasing concentrations of the metal in the media. The

  18. Purification and some properties of a novel maltohexaose-producing exo-amylase from Aerobacter aerogenes.

    PubMed

    Kainuma, K; Wako, K; Kobayashi, S; Nogami, A; Suzuki, S

    1975-12-18

    Maltohexaose producing amylase (EC 3.2.1.-) is the fourth known exo-amylase, the three previously known being glucoamylase, beta-amylase and Pseudomonas stutzeri maltotetraose producing amylase. The enzyme after release from Aerobacter aerogenes cells by 0.1% sodium lauryl sulfate extraction was purified by ammonium sulfate precipitation, DEAE-Sephadex column chromatography and Sephadex G-100 gel filtration to 80-fold of the original sodium lauryl sulfate extract activity, It gave a single band on disc electrophoresis, and the molecular weight by gel filtration was 54 000. This amylase showed maximal activity at 50 degrees C and pH 6.80. The pH stability range was relatively wide, the enzyme retaining more than 90% of its initial activity in the range of 6.50-9.0. 80% of the activity was retained after 15 min at 50 degrees C. This enzyme produced maltohexaose from starch, amylose and amylopectin by exo-attack, but did not act on alpha- or beta-cyclodextrin, pullulan or maltohexaitol. Also the enzyme acted on beta-limit dextrins of amylopectin and glycogen to form branched oligosaccharides. The unusual reaction of this enzyme on beta-limit dextrin is discussed from the standpoint of the stereochemistry of 1,4-alpha- and 1,6-alpha-glucosidic bonds. This is the anomalous amylase for which it is recognized that 1,6-alpha-glucosidic linkages in the substrates can mimic the effect of 1,4-alpha-bonds, as previously observed in pseudo-priming reactions of E. coli phosphorylase. PMID:1094

  19. ?-Amylase from Starchless Seeds of Trigonella Foenum-Graecum and Its Localization in Germinating Seeds

    PubMed Central

    Srivastava, Garima; Kayastha, Arvind M.

    2014-01-01

    Fenugreek (Trigonella foenum-graecum) seeds do not contain starch as carbohydrate reserve. Synthesis of starch is initiated after germination. A ?-amylase from ungerminated fenugreek seeds was purified to apparent electrophoretic homogeneity. The enzyme was purified 210 fold with specific activity of 732.59 units/mg. Mr of the denatured enzyme as determined from SDS-PAGE was 58 kD while that of native enzyme calculated from size exclusion chromatography was 56 kD. Furthermore, its identity was confirmed to be ?-amylase from MALDI-TOF analysis. The optimum pH and temperature was found to be 5.0 and 50°C, respectively. Starch was hydrolyzed at highest rate and enzyme showed a Km of 1.58 mg/mL with it. Antibodies against purified Fenugreek ?-amylase were generated in rabbits. These antibodies were used for localization of enzyme in the cotyledon during different stages of germination using fluorescence and confocal microscopy. Fenugreek ?-amylase was found to be the major starch degrading enzyme depending on the high amount of enzyme present as compared to ?-amylase and also its localization at the periphery of amyloplasts. A new finding in terms of its association with protophloem was observed. Thus, this enzyme appears to be important for germination of seeds. PMID:24551136

  20. Co-culture system of human salivary gland epithelial cells and immune cells from primary Sjögren's syndrome patients: an in vitro approach to study the effects of Rituximab on the activation of the Raf-1/ERK1/2 pathway.

    PubMed

    Lisi, Sabrina; Sisto, Margherita; D'Amore, Massimo; Lofrumento, Dario Domenico

    2015-04-01

    Primary Sjögren's syndrome (pSS) is a chronic autoimmune disorder of the exocrine glands with associated lymphocytic infiltrates in the affected glands. Dryness of the mouth and eyes results from involvement of the salivary and lacrimal glands. The efficacy of Rituximab (RTX) in pSS is still open to debate. This study delineates the signaling pathway involved in RTX-mediated down-regulation of pro-inflammatory factors in a co-culture system of pSS salivary gland epithelial cells (SGEC) with syngeneic pSS B-lymphocytes. In addition, the effects of RTX on the activation of the Raf-1/ERK1/2 pathway in pSS SGEC co-cultured with syngeneic pSS T-lymphocytes were also investigated. This study demonstrated that RTX may interfere with the ERK1/2 pathway in a syngeneic co-culture of pSS SGEC with pSS B-lymphocytes, leading to decreased cytokine production by SGEC. These novel findings reveal that syngeneic co-culture of pSS SGEC with pSS B-lymphocytes leads to a down-regulation of Raf-1 in epithelial cells that adversely regulates the activity of the ERK1/2 pathway and determines a subsequent reduction of the release of pro-inflammatory factors. PMID:25381666

  1. Biological Sciences: Frequencies of Amylase Variants in Drosophila melanogaster

    Microsoft Academic Search

    G. de Jong; A. J. W. Hoorn; G. E. W. Thörig; W. Scharloo

    1972-01-01

    THE selective significance of enzyme polymorphisms remains unsettled1,2. The frequencies of some enzyme variants are correlated with environmental factors3, which might cause frequency changes by acting on closely linked genes. We therefore studied amylase variants in Drosophila melanogaster. A major function of amylase in Drosophila is in the digestion of starch. Genetical and physiological investigations on amylase variants were made

  2. Halophilic alkali- and thermostable amylase from a novel polyextremophilic Amphibacillus sp. NM-Ra2.

    PubMed

    Mesbah, Noha M; Wiegel, Juergen

    2014-09-01

    Extracellular gluco-amylo-pullulanase from Amphibacillus sp. NM-Ra2 was purified to homogeneity by ethanol precipitation, anion exchange chromatography and gel filtration chromatography. Molecular mass of the enzyme was 50kDa (SDS-PAGE). The enzyme showed maximal activity at 1.9 M NaCl, pH50°C 8.0 and 54°C and was active from 0 to 4.3 M NaCl and 37 to 65°C. The enzyme was inhibited by EDTA and was stable and active in the presence of PMSF, DTT, H2O2, Triton-X-100, Tween 20 and Tween 80. Ca2+ is inessential for activity. The amylase was stimulated with K+ and inhibited with Cu2+ and Mg2+. Hg2+, Zn2+ and Fe2+ had no effect on activity. Amylase was stable and active in the presence of ethanol, methanol and benzene (25%, v/v). The enzyme hydrolyzed linear and branched polysaccharides including pullulan, glycogen and amylopectin, and hydrolyzed raw wheat starch and raw corn starch (14.6% and 13.5% over 2 h). Amylase activity was inhibited by soluble starch concentrations greater than 0.3%. The major products of soluble starch hydrolysis were maltose and maltotriose. The amylase, being halophilic and alkali-thermostable, in addition to being resistant to surfactants, oxidizing agents and organic solvents, can find applications in the starch processing, pharmaceutical, food and paper/pulp industries. PMID:25008132

  3. Salivary Defense Proteins: Their Network and Role in Innate and Acquired Oral Immunity

    PubMed Central

    Fábián, Tibor Károly; Hermann, Péter; Beck, Anita; Fejérdy, Pál; Fábián, Gábor

    2012-01-01

    There are numerous defense proteins present in the saliva. Although some of these molecules are present in rather low concentrations, their effects are additive and/or synergistic, resulting in an efficient molecular defense network of the oral cavity. Moreover, local concentrations of these proteins near the mucosal surfaces (mucosal transudate), periodontal sulcus (gingival crevicular fluid) and oral wounds and ulcers (transudate) may be much greater, and in many cases reinforced by immune and/or inflammatory reactions of the oral mucosa. Some defense proteins, like salivary immunoglobulins and salivary chaperokine HSP70/HSPAs (70 kDa heat shock proteins), are involved in both innate and acquired immunity. Cationic peptides and other defense proteins like lysozyme, bactericidal/permeability increasing protein (BPI), BPI-like proteins, PLUNC (palate lung and nasal epithelial clone) proteins, salivary amylase, cystatins, prolin-rich proteins, mucins, peroxidases, statherin and others are primarily responsible for innate immunity. In this paper, this complex system and function of the salivary defense proteins will be reviewed. PMID:22605979

  4. Salivary Gland Cancer: Risk Factors

    MedlinePLUS

    ... Request Permissions Print to PDF Salivary Gland Cancer: Risk Factors Approved by the Cancer.Net Editorial Board , 04/ ... menu on the side of your screen. A risk factor is anything that increases a person’s chance of ...

  5. What Is Salivary Gland Cancer?

    MedlinePLUS

    ... its stage ) are important in predicting outcome. Other rare salivary gland cancers Several other types of cancer ... have a poorer outlook. Epithelial-myoepithelial carcinoma: This rare tumor tends to be low grade, but it ...

  6. Cloning and Expression Analysis of the Bombyx mori ?-amylase Gene (Amy) from the Indigenous Thai Silkworm Strain, Nanglai

    PubMed Central

    Ngernyuang, Nipaporn; Kobayashi, Isao; Promboon, Amornrat; Ratanapo, Sunanta; Tamura, Toshiki; Ngernsiri, Lertluk

    2011-01-01

    ?-Amylase is a common enzyme for hydrolyzing starch. In the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), ?-amylase is found in both digestive fluid and hemolymph. Here, the complete genomic sequence of the Amy gene encoding ?-amylase from a local Thai silkworm, the Nanglai strain, was obtained. This gene was 7981 bp long with 9 exons. The full length Amy cDNA sequence was 1749 bp containing a 1503 bp open reading frame. The ORF encoded 500 amino acid residues. The deduced protein showed 81–54% identity to other insect ?-amylases and more than 50% identity to mammalian enzymes. Southern blot analysis revealed that in the Nanglai strain Amy is a single-copy gene. RT- PCR showed that Amy was transcribed only in the foregut. Transgenic B. mori also showed that the Amy promoter activates expression of the transgene only in the foregut. PMID:21529256

  7. Salivary gland cancer stem cells.

    PubMed

    Adams, April; Warner, Kristy; Nör, Jacques E

    2013-09-01

    Emerging evidence suggests the existence of a tumorigenic population of cancer cells that demonstrate stem cell-like properties such as self-renewal and multipotency. These cells, termed cancer stem cells (CSC), are able to both initiate and maintain tumor formation and progression. Studies have shown that CSC are resistant to traditional chemotherapy treatments preventing complete eradication of the tumor cell population. Following treatment, CSC are able to re-initiate tumor growth leading to patient relapse. Salivary gland cancers are relatively rare but constitute a highly significant public health issue due to the lack of effective treatments. In particular, patients with mucoepidermoid carcinoma or adenoid cystic carcinoma, the two most common salivary malignancies, have low long-term survival rates due to the lack of response to current therapies. Considering the role of CSC in resistance to therapy in other tumor types, it is possible that this unique sub-population of cells is involved in resistance of salivary gland tumors to treatment. Characterization of CSC can lead to better understanding of the pathobiology of salivary gland malignancies as well as to the development of more effective therapies. Here, we make a brief overview of the state-of-the-science in salivary gland cancer, and discuss possible implications of the cancer stem cell hypothesis to the treatment of salivary gland malignancies. PMID:23810400

  8. A lectin from Spatholobus parviflorus inhibits Aspergillus flavus ?-amylase: enzyme kinetics and thermodynamic studies.

    PubMed

    Tintu, Ignatius; Abhilash, Joseph; Dileep, Kalarickal V; Augustine, Anu; Haridas, Madathilkovilakath; Sadasivan, Chittalakkottu

    2014-07-01

    Aspergillus flavus is a commonly found fungal pathogen which produces structurally related and highly toxic secondary metabolites, aflatoxins. It has been proposed that ?-amylase inhibitors may limit the ability of the fungus to produce aflatoxins. Hence, this enzyme is a potent target for the development of antifungal agents. In this study, it was found that Spatholobus parviflorus seed lectin (SPL) can inhibit the growth of A. flavus with a MIC value of 1.5 mg/mL. The enzyme kinetics, molecular modeling and isothermal titration calorimetric studies suggest that SPL can inhibit ?-amylase with Ki value of 0.0042 mm. Hence, it is suggested that the antifungal activity of SPL might be partly due to its ability to inhibit the enzyme ?-amylase. PMID:24460654

  9. Dy3+ and Nd3+ induced genetic mutation of bacillus alpha-amylase.

    PubMed

    Zhang, Dongyan; Hu, Jianhua; Siqin, Bater; Zhang, Tong; Chang, Ying

    2009-07-01

    After treatment with DyCl(3) and NdCl(3), two strains of mutated bacilli showing increased alpha-amylase activity were isolated. The alpha-amylase genes were amplified, cloned, and sequenced. Sequencing revealed that there were either 11 or 14 bases altered in the two genes. These alterations took the form of base substitutions, and transversion was more common than transition. Based on these results, it was concluded that the rare earth compounds DyCl(3) and NdCl(3) were mutagens. PMID:19497621

  10. Enhanced extracellular production of ?-amylase in Bacillus subtilis by optimization of regulatory elements and over-expression of PrsA lipoprotein.

    PubMed

    Chen, Jingqi; Gai, Yuanming; Fu, Gang; Zhou, Wenjuan; Zhang, Dawei; Wen, Jianping

    2015-04-01

    ?-Amylase was used as a heterologous model protein to investigate the effects of promoters, signal peptides and over-expression of an extra-cytoplasmic molecular chaperone, PrsA lipoprotein, on enhancing the secretion of ?-amylase in Bacillus subtilis. Four promoters and six signal peptides were compared, successively, and the highest yield of ?-amylase was achieved under the promotion mediated by PAprE, a strong constitutive promoter, and secretion by SPnprE, a signal peptide from B. subtilis. Moreover, under conditions of overexpressed PrsA lipoprotein, the secretion production and activity of ?-amylase increased to 2.5-fold. The performance of the recombinant B. subtilis 1A751PL31 was evaluated with a fed-batch fermentation in a 7.5 l fermentor. Optimization of regulatory elements and over-expression of PrsA lipoprotein had a significant effect on enhancing the production of ?-amylase in B. subtilis. PMID:25515799

  11. Isolation and characterization of a proteinaceous ?-amylase inhibitor AAI-CC5 from Streptomyces sp. CC5, and its gene cloning and expression.

    PubMed

    Sun, Zhibin; Lu, Weihao; Liu, Pingping; Wang, Hui; Huang, Yan; Zhao, Yuguo; Kong, Yi; Cui, Zhongli

    2015-02-01

    An ?-amylase inhibitor producing Streptomyces sp. strain CC5 was isolated from soil. A proteinaceous ?-amylase inhibitor AAI-CC5 was purified from strain CC5. AAI-CC5 specifically inhibited mammalian ?-amylases. The molecular weight of the inhibitor was determined to be 8,212 Da by MALDI-TOF Mass Spectrum. The N-terminal 15 amino acid residues of the purified AAI-CC5 were DTGSPAPECVEYFQS, which is dissimilar to other reported proteinaceous ?-amylase inhibitors. AAI-CC5 is a pH insensitive and heat-stable protein, and cannot be hydrolysed by trypsin. AAI-CC5 was cloned and expressed in Escherichia coli BL21 (DE3) with a hexa-histidine tag on the C terminal. AAI-CC5 shared 82 % identity with Parvulustat. The recombinant ?-amylase inhibitor was purified to homogeneity by one-step affinity chromatography using Ni(2+)-NTA resin with molecular mass of 9,404 Da. Steady state kinetics studies of ?-amylase and the inhibitor revealed an irreversible, non-competitive inhibition mechanism with IC50 and Ki value of 6.43 ×1 10(-11) and 4.45 × 10(-11) M respectively. These results suggest this novel ?-amylase inhibitor possessed powerful inhibitory activity for ?-amylase, and it may be a candidate in research of diabetes therapy and obesity treatment. PMID:25411086

  12. Autocrine/paracrine dopamine in the salivary glands of the blacklegged tick Ixodes scapularis.

    PubMed

    Ko?i, Juraj; Simo, Ladislav; Park, Yoonseong

    2014-03-01

    Dopamine (DA) is known to be the most potent activator of tick salivary secretion, which is an essential component of successful tick feeding. We examined the quantitative changes of catecholamines using a method coupling high-pressure liquid chromatography with electrochemical detection (HPLC-ECD). We also investigated the levels of catecholamines conjugated to other molecules utilising appropriate methods to hydrolyse the conjugates. Three different biological samples, salivary glands, synganglia, ovaries and haemolymph were compared, and the largest quantity of DA was detected in salivary gland extracts (up to ?100pg/tick), supporting the hypothesis that autocrine/paracrine dopamine activates salivary secretion. Quantitative changes of catecholamines in the salivary glands over the entire blood feeding duration were examined. The amount of dopamine in the salivary glands increased until the day 5 of feeding, at which the rapid engorgement phase began. We also detected a small but significant amount of norepinephrine in the salivary glands. Interestingly, saliva collected after induction of salivary secretion by the cholinergic agonist pilocarpine contained a large amount of DA sulphate with a trace amount of DA, suggesting a potential biological role of DA sulphate in tick saliva. PMID:24503219

  13. SYNERGISTIC ACTION OF BARLEY ALPHA-AMYLASE AND LENTINULA EDODES GLUCOAMYLASE ON RAW STARCH HYDROLYSIS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The genes encoding barley alpha-amylase (AMY1) and Lentinus edodes glucoamylase (GLA) were cloned into Saccharomyces cerevisiae, expressed and constitutively secreted in active forms. The two enzymes were purified by (NH4)2SO4 fractionation and affinity chromatography. AMY1 hydrolyzed starch granul...

  14. Modes of Inhibition of ?-Amylase and ?-Glucosidase by Aqueous Extract of Morinda lucida Benth Leaf

    PubMed Central

    Kazeem, M. I.; Adamson, J. O.; Ogunwande, I. A.

    2013-01-01

    Diabetes mellitus is a metabolic disorder of glucose metabolism. The management of blood glucose level is the hallmark in the treatment of this disease. This may be achieved through the use of oral hypoglycemic drugs such as biguanides, insulin secretagogues, and ?-glucosidase inhibitors. The purpose of the present study was to investigate the inhibitory effect of Morinda lucida leaf extracts on the activities of ?-amylase and ?-glucosidase. This was performed using ?-amylase from Aspergillus oryzae and ?-glucosidase from Saccharomyces cerevisiae. Aqueous extract of Morinda lucida gave the highest percentage yield (9.99%) of the plant out of the three extracts (compared to acetone and ethanolic extracts) and possesses the highest inhibitory activity against ?-amylase (IC50 value of 2.30?mg/mL) and ?-glucosidase (IC50 value of 2.00?mg/mL). Kinetic analysis revealed that the aqueous extract of this plant leaf inhibited the ?-amylase competitively but displayed mixed noncompetitive mode of inhibition towards ?-glucosidase. It can be concluded that aqueous extract of Morinda lucida exhibited the best inhibitory activity on the two enzymes studied and the presence of phytochemicals like flavonoids, saponins, and tannins may have contributed greatly to the inhibitory activity of the plant extract. PMID:24455701

  15. Kinetic Analysis of Amylase Using Quantitative Benedict's and Iodine Starch Reagents

    ERIC Educational Resources Information Center

    Cochran, Beverly; Lunday, Deborah; Miskevich, Frank

    2008-01-01

    Quantitative analysis of carbohydrates is a fundamental analytical tool used in many aspects of biology and chemistry. We have adapted a technique developed by Mathews et al. using an inexpensive scanner and open-source image analysis software to quantify amylase activity using both the breakdown of starch and the appearance of glucose. Breakdown…

  16. Supramolecular interactions mediated thermal stabilization for ?-amylase modified with a ?-cyclodextrin-carboxymethylcellulose polymer

    Microsoft Academic Search

    Rodolfo Darias; Ivelises Herrera; Alex Fragoso; Roberto Cao; Reynaldo Villalonga

    2002-01-01

    a-Amylase from Bacillus subtilis was modified with a ß-cyclodextrin-carboxymethylcellulose polymer and retained 90% of its initial activity. Its thermostability was enhanced from 68 °C to 82.5 °C over 10 min incubation and the resistance to inactivation at 75 °C was increased 5-fold. The influence of supramolecular associations polymer-protein on enzyme thermostabilization was demonstrated.

  17. Effects of alpha-amylase and its inhibitors on acid production from cooked starch by oral streptococci.

    PubMed

    Aizawa, S; Miyasawa-Hori, H; Nakajo, K; Washio, J; Mayanagi, H; Fukumoto, S; Takahashi, N

    2009-01-01

    This study evaluated acid production from cooked starch by Streptococcus mutans, Streptococcus sobrinus, Streptococcus sanguinis and Streptococcus mitis, and the effects of alpha-amylase inhibitors (maltotriitol and acarbose) and xylitol on acid production. Streptococcal cell suspensions were anaerobically incubated with various carbohydrates that included cooked potato starch in the presence or absence of alpha-amylase. Subsequently, the fall in pH and the acid production rate at pH 7.0 were measured. In addition, the effects of adding alpha-amylase inhibitors and xylitol to the reaction mixture were evaluated. In the absence of alpha-amylase, both the fall in pH and the acid production rate from cooked starch were small. On the other hand, in the presence of alpha-amylase, the pH fell to 3.9-4.4 and the acid production rate was 0.61-0.92 micromol per optical density unit per min. These values were comparable to those for maltose. When using cooked starch, the fall in pH by S. sanguinis and S. mitis was similar to that by S. mutans and S. sobrinus. For all streptococci, alpha-amylase inhibitors caused a decrease in acid production from cooked starch, although xylitol only decreased acid production by S. mutans and S. sobrinus. These results suggest that cooked starch is potentially acidogenic in the presence of alpha-amylase, which occurs in the oral cavity. In terms of the acidogenic potential of cooked starch, S. sanguinis and S. mitis were comparable to S. mutans and S. sobrinus. Alpha-amylase inhibitors and xylitol might moderate this activity. PMID:19136828

  18. Tobacco Plants Transformed with the Bean ?ai Gene Express an Inhibitor of Insect ?-Amylase in Their Seeds 1

    PubMed Central

    Altabella, Teresa; Chrispeels, Maarten J.

    1990-01-01

    Bean (Phaseolus vulgaris L.) seeds contain a putative plant defense protein that inhibits insect and mammalian but not plant ?-amylases. We recently (J Moreno, MJ Chrispeels [1989] Proc Natl Acad Sci USA 86:7885-7889) presented strong circumstantial evidence that this ?-amylase inhibitor (?AI) is encoded by an already-identified lectin gene whose product is referred to as lectin-like-protein (LLP). We have now made a chimeric gene consisting of the coding sequence of the lectin gene that encodes LLP and the 5? and 3? flanking sequences of the lectin gene that encodes phytohemagglutinin-L. When this chimeric gene was expressed in transgenic tobacco (Nicotiana tabacum), we observed in the seeds a series of polypeptides (Mr 10,000-18,000) that cross-react with antibodies to the bean ?-amylase inhibitor. Most of these polypeptides bind to a pig pancreas ?-amylase affinity column. An extract of the seeds of the transformed tobacco plants inhibits pig pancreas ?-amylase activity as well as the ?-amylase present in the midgut of Tenebrio molitor. We suggest that introduction of this lectin gene (to be called ?ai) into other leguminous plants may be a strategy to protect the seeds from the seed-eating larvae of Coleoptera. Images Figure 2 Figure 3 Figure 5 PMID:16667540

  19. Gene cloning and characterization of an ?-amylase from Alteromonas macleodii B7 for Enteromorpha polysaccharide degradation.

    PubMed

    Han, Xuefeng; Lin, Bokun; Ru, Ganji; Zhang, Zhibiao; Liu, Yan; Hu, Zhong

    2014-02-28

    Enteromorpha polysaccharides (EP) extracted from green algae have displayed a wide variety of biological activities. However, their high molecular weight leads to a high viscosity and low solubility, and therefore, greatly restrains their application. To solve this problem, bacteria from the surface of Enteromorpha were screened, and an Alteromonas macleodii strain B7 was found to be able to decrease the molecular weight of EP in culture media. Proteins harvested from the supernatant of the A. macleodii B7 culture were subjected to native gel electrophoresis, and a band corresponding to the Enteromorpha polysaccharide lyase (EPL) was detected by activity staining. The enzyme identity was subsequently confirmed by MALDI-TOF/TOF mass spectrometry as the putative ?-amylase reported in A. macleodii ATCC 27126. The amylase gene (amySTU) from A. macleodii B7 was cloned into Escherichia coli, resulting in highlevel expression of the recombinant enzyme with EP-degrading activity. AmySTU was found to be cold-adapted; however, its optimal enzyme activity was detected at 40°C. The ?-amylase was highly stable over a broad pH range (5.5-10) with the optimal pH at 7.5-8.0. The highest enzyme activity was detected when NaCl concentration was 2%, which dropped by 50% when the NaCl concentration was increased to 16%, showing an excellent nature of halotolerance. Furthermore, the amylase activity was not significantly affected by tested surfactants or the presence of some organic solvents. Therefore, the A. macleodii strain B7 and its ?-amylase can be useful in lowering EP molecular weight and in starch processing. PMID:24262656

  20. Antidiabetic Indian Plants: A Good Source of Potent Amylase Inhibitors

    PubMed Central

    Bhat, Menakshi; Zinjarde, Smita S.; Bhargava, Shobha Y.; Kumar, Ameeta Ravi; Joshi, Bimba N.

    2011-01-01

    Diabetes is known as a multifactorial disease. The treatment of diabetes (Type II) is complicated due to the inherent patho-physiological factors related to this disease. One of the complications of diabetes is post-prandial hyperglycemia (PPHG). Glucosidase inhibitors, particularly ?-amylase inhibitors are a class of compounds that helps in managing PPHG. Six ethno-botanically known plants having antidiabetic property namely, Azadirachta indica Adr. Juss.; Murraya koenigii (L.) Sprengel; Ocimum tenuflorum (L.) (syn: Sanctum); Syzygium cumini (L.) Skeels (syn: Eugenia jambolana); Linum usitatissimum (L.) and Bougainvillea spectabilis were tested for their ability to inhibit glucosidase activity. The chloroform, methanol and aqueous extracts were prepared sequentially from either leaves or seeds of these plants. It was observed that the chloroform extract of O. tenuflorum; B. spectabilis; M. koenigii and S. cumini have significant ?-amylase inhibitory property. Plants extracts were further tested against murine pancreatic, liver and small intestinal crude enzyme preparations for glucosidase inhibitory activity. The three extracts of O. tenuflorum and chloroform extract of M. koenigi showed good inhibition of murine pancreatic and intestinal glucosidases as compared with acarbose, a known glucosidase inhibitor. PMID:18955350

  1. ?-Amylase-assisted extraction of polysaccharides from Panax ginseng.

    PubMed

    Sun, Lin; Wu, Di; Ning, Xin; Yang, Guang; Lin, Ziheng; Tian, Meihong; Zhou, Yifa

    2015-04-01

    In this paper, ?-amylase-assisted extraction was used to isolate the polysaccharide that remained in hot water-extracted ginseng. The yield of the polysaccharide was 9.0%, almost equal to that of the hot water-extracted polysaccharide. Using anion exchange and gel permeation chromatography, the polysaccharide was fractionated into a neutral polysaccharide fraction and six pectic fractions. The neutral fraction accounted for 76% of the polysaccharide and contained both amylopectin and amylose. The pectic polysaccharide fractions were identified to be arabinogalactan, type-I rhamnogalacturonan and homogalacturonan-type pectin by high-performance liquid chromatography, Fourier transform-infrared and nuclear magnetic resonance analysis. Structural and lymphocyte proliferation activity results showed that these polysaccharides were different from those extracted by hot water, indicating that ginseng contains complex polysaccharides with diverse structures, which results in its diverse pharmacological activities. The ?-amylase-assisted extraction is a novel method for preparing ginseng polysaccharides and could be applied toward the further study and exploration of ginseng. These findings provide technical and theoretical support for ginseng pharmacology. PMID:25616118

  2. ?-Amylase inhibitor-1 gene from Phaseolus vulgaris expressed in Coffea arabica plants inhibits ?-amylases from the coffee berry borer pest

    PubMed Central

    2010-01-01

    Background Coffee is an important crop and is crucial to the economy of many developing countries, generating around US$70 billion per year. There are 115 species in the Coffea genus, but only two, C. arabica and C. canephora, are commercially cultivated. Coffee plants are attacked by many pathogens and insect-pests, which affect not only the production of coffee but also its grain quality, reducing the commercial value of the product. The main insect-pest, the coffee berry borer (Hypotheneumus hampei), is responsible for worldwide annual losses of around US$500 million. The coffee berry borer exclusively damages the coffee berries, and it is mainly controlled by organochlorine insecticides that are both toxic and carcinogenic. Unfortunately, natural resistance in the genus Coffea to H. hampei has not been documented. To overcome these problems, biotechnological strategies can be used to introduce an ?-amylase inhibitor gene (?-AI1), which confers resistance against the coffee berry borer insect-pest, into C. arabica plants. Results We transformed C. arabica with the ?-amylase inhibitor-1 gene (?-AI1) from the common bean, Phaseolus vulgaris, under control of the seed-specific phytohemagglutinin promoter (PHA-L). The presence of the ?-AI1 gene in six regenerated transgenic T1 coffee plants was identified by PCR and Southern blotting. Immunoblotting and ELISA experiments using antibodies against ?-AI1 inhibitor showed a maximum ?-AI1 concentration of 0.29% in crude seed extracts. Inhibitory in vitro assays of the ?-AI1 protein against H. hampei ?-amylases in transgenic seed extracts showed up to 88% inhibition of enzyme activity. Conclusions This is the first report showing the production of transgenic coffee plants with the biotechnological potential to control the coffee berry borer, the most important insect-pest of crop coffee. PMID:20565807

  3. Identification of Regulatory Factors for Mesenchymal Stem Cell-Derived Salivary Epithelial Cells in a Co-Culture System

    PubMed Central

    Park, Yun-Jong; Koh, Jin; Gauna, Adrienne E.; Chen, Sixue; Cha, Seunghee

    2014-01-01

    Patients with Sjögren’s syndrome or head and neck cancer patients who have undergone radiation therapy suffer from severe dry mouth (xerostomia) due to salivary exocrine cell death. Regeneration of the salivary glands requires a better understanding of regulatory mechanisms by which stem cells differentiate into exocrine cells. In our study, bone marrow-derived mesenchymal stem cells were co-cultured with primary salivary epithelial cells from C57BL/6 mice. Co-cultured bone marrow-derived mesenchymal stem cells clearly resembled salivary epithelial cells, as confirmed by strong expression of salivary gland epithelial cell-specific markers, such as alpha-amylase, muscarinic type 3 receptor, aquaporin-5, and cytokeratin 19. To identify regulatory factors involved in this differentiation, transdifferentiated mesenchymal stem cells were analyzed temporarily by two-dimensional-gel-electrophoresis, which detected 58 protein spots (>1.5 fold change, p<0.05) that were further categorized into 12 temporal expression patterns. Of those proteins only induced in differentiated mesenchymal stem cells, ankryin-repeat-domain-containing-protein 56, high-mobility-group-protein 20B, and transcription factor E2a were selected as putative regulatory factors for mesenchymal stem cell transdifferentiation based on putative roles in salivary gland development. Induction of these molecules was confirmed by RT-PCR and western blotting on separate sets of co-cultured mesenchymal stem cells. In conclusion, our study is the first to identify differentially expressed proteins that are implicated in mesenchymal stem cell differentiation into salivary gland epithelial cells. Further investigation to elucidate regulatory roles of these three transcription factors in mesenchymal stem cell reprogramming will provide a critical foundation for a novel cell-based regenerative therapy for patients with xerostomia. PMID:25402494

  4. Identification of regulatory factors for mesenchymal stem cell-derived salivary epithelial cells in a co-culture system.

    PubMed

    Park, Yun-Jong; Koh, Jin; Gauna, Adrienne E; Chen, Sixue; Cha, Seunghee

    2014-01-01

    Patients with Sjögren's syndrome or head and neck cancer patients who have undergone radiation therapy suffer from severe dry mouth (xerostomia) due to salivary exocrine cell death. Regeneration of the salivary glands requires a better understanding of regulatory mechanisms by which stem cells differentiate into exocrine cells. In our study, bone marrow-derived mesenchymal stem cells were co-cultured with primary salivary epithelial cells from C57BL/6 mice. Co-cultured bone marrow-derived mesenchymal stem cells clearly resembled salivary epithelial cells, as confirmed by strong expression of salivary gland epithelial cell-specific markers, such as alpha-amylase, muscarinic type 3 receptor, aquaporin-5, and cytokeratin 19. To identify regulatory factors involved in this differentiation, transdifferentiated mesenchymal stem cells were analyzed temporarily by two-dimensional-gel-electrophoresis, which detected 58 protein spots (>1.5 fold change, p<0.05) that were further categorized into 12 temporal expression patterns. Of those proteins only induced in differentiated mesenchymal stem cells, ankryin-repeat-domain-containing-protein 56, high-mobility-group-protein 20B, and transcription factor E2a were selected as putative regulatory factors for mesenchymal stem cell transdifferentiation based on putative roles in salivary gland development. Induction of these molecules was confirmed by RT-PCR and western blotting on separate sets of co-cultured mesenchymal stem cells. In conclusion, our study is the first to identify differentially expressed proteins that are implicated in mesenchymal stem cell differentiation into salivary gland epithelial cells. Further investigation to elucidate regulatory roles of these three transcription factors in mesenchymal stem cell reprogramming will provide a critical foundation for a novel cell-based regenerative therapy for patients with xerostomia. PMID:25402494

  5. Extraction and purification of beta-amylase from stems of Abrus precatorius by three phase partitioning.

    PubMed

    Sagu, Sorel Tchewonpi; Nso, Emmanuel Jong; Homann, Thomas; Kapseu, César; Rawel, Harshadrai M

    2015-09-15

    The stems of Abrus precatorius were used to extract a beta-amylase enriched fraction. A three phase partitioning method and a Doehlert design with 3 variables (ratio of crude extract/t-butanol, the ammonium sulphate saturation and pH) were used. The data was fitted in a second-order polynomial model and the parameters were optimized to enrich beta-amylase. Experimental responses for the modulation were recovery of activity and the purification factor. The optimal conditions were: a ratio of crude extract/t-butanol of 0.87 (v/v), saturation in ammonium sulphate of 49.46% (w/v) and a pH of 5.2. An activity recovery of 156.2% and a purification factor of 10.17 were found. The enriched enzyme was identified as a beta-amylase and its molecular weight was 60.1kDa. Km and Vmax values were 79.37mg/ml and 5.13U/ml, respectively and the highest activity was registered at a temperature of 70°C and a pH between 6 and 6.5. A significant stabilization of the beta-amylase was observed up to 65°C. PMID:25863622

  6. Structural genes encoding the thermophilic alpha-amylases of Bacillus stearothermophilus and Bacillus licheniformis.

    PubMed Central

    Gray, G L; Mainzer, S E; Rey, M W; Lamsa, M H; Kindle, K L; Carmona, C; Requadt, C

    1986-01-01

    The genes encoding the thermostable alpha-amylases of Bacillus stearothermophilus and B. licheniformis were cloned in Escherichia coli, and their DNA sequences were determined. The coding and deduced polypeptide sequences are 59 and 62% homologous to each other, respectively. The B. stearothermophilus protein differs most significantly from that of B. licheniformis in that it possesses a 32-residue COOH-terminal tail. Transformation of E. coli with vectors containing either gene resulted in the synthesis and secretion of active enzymes similar to those produced by the parental organisms. A plasmid was constructed in which the promoter and the NH2-terminal two-thirds of the B. stearothermophilus coding sequence was fused out of frame to the entire mature coding sequence of the B. licheniformis gene. Approximately 1 in 5,000 colonies transformed with this plasmid was found to secrete an active amylase. Hybridization analysis of plasmids isolated from these amylase-positive colonies indicated that the parental coding sequences had recombined by homologous recombination. DNA sequence analysis of selected hybrid genes revealed symmetrical, nonrandom distribution of loci at which the crossovers had resolved. Several purified hybrid alpha-amylases were characterized and found to differ with respect to thermostability and specific activity. PMID:3009417

  7. Effects of radiation on parotid salivary function

    Microsoft Academic Search

    James E. Marks; Catherine C. Davis; Vikki L. Gottsman; James E. Purdy; Fransiska Lee

    1981-01-01

    Postoperative electron beam irradiation of patients with parotid cancer has been used regularly at the Mallinckrodt Institute of Radiology to spare the opposite parotid and to preserve salivary function. Only anecdotal reports of amount of radiation required to ablate salivary function exist. To establish a dose-response curve for the human parotid, selective measurements of right and left parotid salivary flow

  8. Salivary gland calculi – contemporary methods of imaging

    PubMed Central

    Rzymska-Grala, Iwona; Stopa, Zygmunt; Grala, Bart?omiej; Go??biowski, Marek; Wanyura, Hubert; Zuchowska, Anna; Sawicka, Monika; Zmorzy?ski, Micha?

    2010-01-01

    Summary Sialolithiasis is the most common disorder of major salivary glands. The main site of salivary stones’ formation is submandibular gland, followed by parotid and sublingual gland. The aim of this article was to present current diagnostic imaging modalities carried out in patients suspected with salivary stones on the basis of own material and review of literature. Current diagnostic imaging tools used in the imaging of salivary stones were described and illustrated in this paper. These are: conventional radiography, sialography, ultrasonography, computed tomography, magnetic resonance sialography and sialoendoscopy. Digital subtraction sialography and ultrasonography are the methods of choice in the imaging of salivary gland calculi. Although sialography is a very old diagnostic method, still it is the best diagnostic tool in the imaging of subtle anatomy of salivary gland duct system. Digital subtraction sialography can show the exact location of salivary stone and enables imaging of salivary ducts’ pathology (e.g. stenoses), which is especially important when sialoendoscopy is planned. Sialography is also used as the treatment method, i.e. interventional sialography. Nonenhanced computed tomography is recommended when multiple and tiny salivary stones are suspected. Magnetic resonance imaging is the evolving alternative diagnostic method. In this diagnostic modality there is no need for salivary ducts’ cannulation and administration of contrast material. Thus magnetic resonance sialography can also be carried out in the acute sialoadenitis. In the future, sialoendoscopy may become one of the main diagnostic and treatment procedures for salivary duct disorders, especially in salivary stone cases. PMID:22802788

  9. Purification, Characterization, and Potential of Saline Waste Water Remediation of a Polyextremophilic ?-Amylase from an Obligate Halophilic Aspergillus gracilis

    PubMed Central

    Ali, Imran; Akbar, Ali; Yanwisetpakdee, Benjawan; Prasongsuk, Sehanat; Lotrakul, Pongtharin; Punnapayak, Hunsa

    2014-01-01

    An obligate halophilic Aspergillus gracilis which was isolated from a hypersaline man-made saltern from Thailand was screened for its potential of producing extracellular ?-amylase in the previous studies. In this study the ?-amylase was extracted and purified by the help of column chromatography using Sephadex G-100 column. Presence of amylase was verified by SDS-PAGE analysis, showing a single band of approximately 35?kDa. The specific activity of the enzyme was found to be 131.02?U/mg. The Lineweaver-Burk plot showed the Vmax and Km values of 8.36?U/mg and 6.33?mg/mL, respectively. The enzyme was found to have the best activity at 5 pH, 60°C, and 30% of NaCl concentration, showing its polyextremophilic nature. The use of various additives did not show much variation in the activity of enzyme, showing its resilience against inhibitors. The enzyme, when tested for its use for synthetic waste water remediation by comparing its activity with commercial amylase in different salt concentrations showed that the ?-amylase from A. gracilis was having better performance at increasing salt concentrations than the commercial one. This shows its potential to be applied in saline waste water and other low water activity effluents for bioremediation. PMID:24949415

  10. Radiation-Induced Salivary Gland Dysfunction Results From p53-Dependent Apoptosis

    SciTech Connect

    Avila, Jennifer L. [Department of Physiological Sciences, University of Arizona, Tucson, AZ (United States); Grundmann, Oliver; Burd, Randy [Department of Nutritional Sciences, University of Arizona, Tucson, AZ (United States); Limesand, Kirsten H. [Department of Physiological Sciences, University of Arizona, Tucson, AZ (United States); Department of Nutritional Sciences, University of Arizona, Tucson, AZ (United States)], E-mail: limesank@u.arizona.edu

    2009-02-01

    Purpose: Radiotherapy for head-and-neck cancer causes adverse secondary side effects in the salivary glands and results in diminished quality of life for the patient. A previous in vivo study in parotid salivary glands demonstrated that targeted head-and-neck irradiation resulted in marked increases in phosphorylated p53 (serine{sup 18}) and apoptosis, which was suppressed in transgenic mice expressing a constitutively active mutant of Akt1 (myr-Akt1). Methods and Materials: Transgenic and knockout mouse models were exposed to irradiation, and p53-mediated transcription, apoptosis, and salivary gland dysfunction were analyzed. Results: The proapoptotic p53 target genes PUMA and Bax were induced in parotid salivary glands of mice at early time points after therapeutic radiation. This dose-dependent induction requires expression of p53 because no radiation-induced expression of PUMA and Bax was observed in p53-/- mice. Radiation also induced apoptosis in the parotid gland in a dose-dependent manner, which was p53 dependent. Furthermore, expression of p53 was required for the acute and chronic loss of salivary function after irradiation. In contrast, apoptosis was not induced in p53-/- mice, and their salivary function was preserved after radiation exposure. Conclusions: Apoptosis in the salivary glands after therapeutic head-and-neck irradiation is mediated by p53 and corresponds to salivary gland dysfunction in vivo.

  11. [Salivary gland tumors in children].

    PubMed

    Thariat, Juliette; Vedrine, Pierre-Olivier; Orbach, Daniel; Marcy, Pierre-Yves; Badoual, Cécile; Butori, Catherine; Teissier, Natacha; Toussaint, Bruno; Castillo, Laurent

    2011-07-01

    Salivary gland tumors in children are rare: they correspond to 8-10% of head and neck pediatric tumors. Clinicians of all disciplines should be aware of this diagnosis in front of non-inflammatory mass of the parotid or in the territory of other salivary glands. In children, 50% of salivary gland tumors are malignant which contrasts with a 10-25% risk in adults. Epithelial tumors are the most common, mucoepidermoïd carcinomas of the parotid in particular. Surgery is the treatment of choice in epithelial tumors. Adjuvant radiotherapy may be indicated in case of unfavorable prognostic factors but must be balanced with the risk of radiation-induced growth defects and secondary cancer. The role of chemotherapy is limited in these tumors, but should be discussed in case of an inoperable or metastatic lesion. PMID:21690035

  12. Human mesenchymal stem cells cultured with salivary gland biopsies adopt an epithelial phenotype.

    PubMed

    Maria, Ola M; Tran, Simon D

    2011-06-01

    Sjogren's syndrome and radiotherapy for head and neck cancer result in severe xerostomia and irreversible salivary gland damage for which no effective treatment is currently available. Cell culture methods of primary human salivary gland epithelial cells (huSGs) are slow and cannot provide a sufficient number of cells. In addition, the majority of cultured huSGs are of a ductal phenotype and thus not fluid/saliva secretory cells. Some reports indicated that mesenchymal stem cells (MSCs) possessed the potential to differentiate into epithelial cells. To test this hypothesis with huSGs, a coculture system containing 2 chambers separated by a polyester membrane was used to study the capacity of human MSCs to adopt an epithelial phenotype when cocultured with human salivary gland biopsies. Results were that 20%-40% of cocultured MSCs expressed tight junction proteins [claudin-1 (CLDN-1), -2, -3, and -4; occludin; junctional adhesion molecule-A; and zonula occludens-1] as well as other epithelial markers [aquaporin-5, ?-amylase (?-AMY), and E-cadherin], and generated a higher transepithelial electrical resistance. Electron microscopy demonstrated that these MSCs had comparable cellular structures to huSGs, such as tight junction structures and numerous secretory granules. Quantitative real time (RT)-polymerase chain reaction revealed an upregulation of several salivary genes (aquaporin-5, AMY, and CLDN-2). Moreover, the amounts of ?-AMY detected in cocultured MSCs were comparable to those detected in huSGs control cultures. These data suggest that cocultured MSCs can demonstrate a temporary change into a salivary gland acinar phenotype. PMID:21187001

  13. Purification and Characterization of a Highly Efficient Calcium-Independent ?-Amylase from Talaromyces pinophilus 1-95

    PubMed Central

    Xian, Liang; Wang, Fei; Luo, Xiang; Feng, Yu-Liang; Feng, Jia-Xun

    2015-01-01

    Alpha-amylase is a very important enzyme in the starch conversion process. Most of the ?-amylases are calcium-dependent and exhibit poor performance in the simultaneous saccharification and fermentation process of industrial bioethanol production that uses starch as feedstock. In this study, an extracellular amylolytic enzyme was purified from the culture broth of newly isolated Talaromyces pinophilus strain 1-95. The purified amylolytic enzyme, with an apparent molecular weight of 58 kDa on SDS-PAGE, hydrolyzed maltopentaose, maltohexaose, and maltoheptaose into mainly maltose and maltotriose and minor amount of glucose, confirming the endo-acting mode of the enzyme, and hence, was named Talaromyces pinophilus ?-amylase (TpAA). TpAA was most active at pH 4.0–5.0 (with the temperature held at 37°C) and 55°C (at pH 5.0), and stable within the pH range of 5.0–9.5 (at 4°C) and below 45°C (at pH 5.0). Interestingly, the Ca2+ did not improve its enzymatic activity, optimal temperature, or thermostability of the enzyme, indicating that the TpAA was Ca2+-independent. TpAA displayed higher enzyme activity toward malto-oligosaccharides and dextrin than other previously reported ?-amylases. This highly active Ca2+-independent ?-amylase may have potential applications in starch-to-ethanol conversion process. PMID:25811759

  14. Crystal structures of the psychrophilic alpha-amylase from Alteromonas haloplanctis in its native form and complexed with an inhibitor.

    PubMed Central

    Aghajari, N.; Feller, G.; Gerday, C.; Haser, R.

    1998-01-01

    Alteromonas haloplanctis is a bacterium that flourishes in Antarctic sea-water and it is considered as an extreme psychrophile. We have determined the crystal structures of the alpha-amylase (AHA) secreted by this bacterium, in its native state to 2.0 angstroms resolution as well as in complex with Tris to 1.85 angstroms resolution. The structure of AHA, which is the first experimentally determined three-dimensional structure of a psychrophilic enzyme, resembles those of other known alpha-amylases of various origins with a surprisingly greatest similarity to mammalian alpha-amylases. AHA contains a chloride ion which activates the hydrolytic cleavage of substrate alpha-1,4-glycosidic bonds. The chloride binding site is situated approximately 5 angstroms from the active site which is characterized by a triad of acid residues (Asp 174, Glu 200, Asp 264). These are all involved in firm binding of the Tris moiety. A reaction mechanism for substrate hydrolysis is proposed on the basis of the Tris inhibitor binding and the chloride activation. A trio of residues (Ser 303, His 337, Glu 19) having a striking spatial resemblance with serine-protease like catalytic triads was found approximately 22 angstroms from the active site. We found that this triad is equally present in other chloride dependent alpha-amylases, and suggest that it could be responsible for autoproteolytic events observed in solution for this cold adapted alpha-amylase. PMID:9541387

  15. Purification and Characterization of a Highly Efficient Calcium-Independent ?-Amylase from Talaromyces pinophilus 1-95.

    PubMed

    Xian, Liang; Wang, Fei; Luo, Xiang; Feng, Yu-Liang; Feng, Jia-Xun

    2015-01-01

    Alpha-amylase is a very important enzyme in the starch conversion process. Most of the ?-amylases are calcium-dependent and exhibit poor performance in the simultaneous saccharification and fermentation process of industrial bioethanol production that uses starch as feedstock. In this study, an extracellular amylolytic enzyme was purified from the culture broth of newly isolated Talaromyces pinophilus strain 1-95. The purified amylolytic enzyme, with an apparent molecular weight of 58 kDa on SDS-PAGE, hydrolyzed maltopentaose, maltohexaose, and maltoheptaose into mainly maltose and maltotriose and minor amount of glucose, confirming the endo-acting mode of the enzyme, and hence, was named Talaromyces pinophilus ?-amylase (TpAA). TpAA was most active at pH 4.0-5.0 (with the temperature held at 37°C) and 55°C (at pH 5.0), and stable within the pH range of 5.0-9.5 (at 4°C) and below 45°C (at pH 5.0). Interestingly, the Ca2+ did not improve its enzymatic activity, optimal temperature, or thermostability of the enzyme, indicating that the TpAA was Ca2+-independent. TpAA displayed higher enzyme activity toward malto-oligosaccharides and dextrin than other previously reported ?-amylases. This highly active Ca2+-independent ?-amylase may have potential applications in starch-to-ethanol conversion process. PMID:25811759

  16. Expression of ?-amylase inhibitors in diploid Triticum species.

    PubMed

    Zoccatelli, Gianni; Sega, Michela; Bolla, Michela; Cecconi, Daniela; Vaccino, Patrizia; Rizzi, Corrado; Chignola, Roberto; Brandolini, Andrea

    2012-12-15

    The aim of the work was to characterize the expression of various ?-amylase inhibitors (?AIs), well known anti-nutritional compounds, for the development of healthier diploid wheat-based functional foods. The salt-soluble protein fractions from the seeds of 53 accessions among Triticum monococcum subsp. monococcum (T.m.), T. monococcum subsp. boeoticum (T.b.) and Triticum urartu (T.u.) were analyzed by immunoblotting after SDS-PAGE and Urea-PAGE using polyclonal antibodies (PABs) raised against 0.19 and 0.28 ?AIs expressed in bread-wheat. Reverse zymography with human saliva and Tenebrio molitor ?-amylases was used to assay inhibition activity. A great variability of the expression of ?AI-related proteins was observed among T.b. and T.u. PABs, and reverse zymography revealed different bands, often not correlating with those present in bread-wheat. Two-dimensional electrophoresis followed by immunoblotting and mass spectrometric analysis identified these proteins as ?AIs. Interestingly, no signal was observed within T.m. accessions. This makes T.m. an important candidate for the production of novel functional foods. PMID:22980853

  17. Salivary cortisol determination in patients from the Basque Country with recurrent aphthous stomatitis. A pilot study

    PubMed Central

    Martínez-Conde-Llamosas, Rafael; López-Vicente, José; Uribarri-Etxebarria, Agurne; Aguirre-Urizar, José M.

    2013-01-01

    Objectives: Stress and anxiety are controversial factors involved in the complex pathogenesis of Recurrent Aphthous Stomatitis (RAS). The determination of salivary cortisol is a useful, simple and safe test to detect states of high stress or anxiety. The aim of this study is to check for changes in salivary cortisol levels in patients with RAS during periods of active disease. Study design: A measurement of cortisol employing Enzyme-Linked Immuno Sorbent Assay (ELISA) was carried out in samples of unstimulated saliva from 20 patients with active lesions of RAS and 10 healthy individuals used as controls. Results: Increased levels of salivary cortisol were detected in 3 cases, all of them within the group of patients with RAS. In none of the control group patients the level of salivary cortisol was increased. The mean level of salivary cortisol was 0.64 mg / dl (range 0.2 to 1.62) for patients with RAS and 0.57 mg / dl (range 0.25 to 1.09) for controls. Conclusion: Salivary cortisol levels are not statistically higher in patients with active lesions of RAS. Key words:Recurrent aphthous stomatitis, cortisol, oral ulcers, canker sores, salivary cortisol. PMID:23385495

  18. 21 CFR 184.1012 - ?-Amylase enzyme preparation from Bacillus stearothermophilus.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...2009-04-01 true α-Amylase enzyme preparation from Bacillus stearothermophilus...as GRAS § 184.1012 ?-Amylase enzyme preparation from Bacillus stearothermophilus. (a) ?-Amylase enzyme preparation is obtained from...

  19. Solid State Fermentation of a Raw Starch Digesting Alkaline Alpha-Amylase from Bacillus licheniformis RT7PE1 and Its Characteristics

    PubMed Central

    Tabassum, Romana; Khaliq, Shazia; Rajoka, Muhammad Ibrahim; Agblevor, Foster

    2014-01-01

    The thermodynamic and kinetic properties of solids state raw starch digesting alpha amylase from newly isolated Bacillus licheniformis RT7PE1 strain were studied. The kinetic values Qp, Yp/s, Yp/X, and qp were proved to be best with 15% wheat bran. The molecular weight of purified enzyme was 112?kDa. The apparent Km and Vmax values for starch were 3.4?mg mL?1 and 19.5?IU?mg?1 protein, respectively. The optimum temperature and pH for ?-amylase were 55°C, 9.8. The half-life of enzyme at 95°C was 17h. The activation and denaturation activation energies were 45.2 and 41.2?kJ mol?1, respectively. Both enthalpies (?H?) and entropies of activation (?S?) for denaturation of ?-amylase were lower than those reported for other thermostable ?-amylases. PMID:24587909

  20. Antibacterial Activity of the Purified Peroxidase from Human Parotid Saliva

    PubMed Central

    Slowey, R. R.; Eidelman, S.; Klebanoff, S. J.

    1968-01-01

    The peroxidase of human parotid saliva has been purified by concentration, gel filtration on Sephadex G-200, and ion exchange chromatography on Amberlite CG-50. The purified product was devoid of amylase activity, lysozyme activity, and immunoglobulin A (IgA). However, it had an inhibitory effect on the growth of Lactobacillus acidophilus in complete growth medium and on lysine accumulation by L. acidophilus in a buffer-glucose medium, when combined with thiocyanate ions. The concentrations of peroxidase and thiocyanate ions employed were within the range found in saliva. The fractions which contained IgA did not have an anti-bacterial effect on L. acidophilus under the conditions employed. Parotid saliva also contained low molecular weight inhibitors of peroxidase activity. These studies support the involvement of the salivary peroxidase in an antibacterial system in saliva. PMID:4183966

  1. Segments of amino acid sequence similarity in beta-amylases.

    PubMed

    Friedberg, F; Rhodes, C

    1988-01-01

    In alpha-amylases from animals, plants and bacteria and in beta-amylases from plants and bacteria a number of segments exhibit amino acid sequence similarity specific to the alpha or to the beta type, respectively. In the case of the beta-amylases the similar sequence regions are extensive and they are disrupted only by short interspersed dissimilar regions. Close to the C terminus, however, no such sequence similarity exist. PMID:2464171

  2. Does Drainage Fluid Amylase Reflect Pancreatic Leakage after Pancreaticoduodenectomy?

    Microsoft Academic Search

    Yi-Ming Shyr; Cheng-Hsi Su; Chew-Wun Wu; Wing-Yiu Lui

    2003-01-01

      Abstract\\u000a \\u000a This study tried to determine if drainage fluid amylase reflects pancreatic leakage after pancreaticoduodenectomy and to determine\\u000a the factors affecting the drainage amylase level. Patients undergoing pancreaticoduodenectomy were recruited. The drainage\\u000a amylase was measured from postoperative day (POD) 1 to POD 7. Direct evidence of pancreatic leakage was provided by upper\\u000a gastrointestinal studies using a water-soluble contrast medium and

  3. Effects of cigarette smoke on salivary protein tyrosine nitration

    PubMed Central

    2010-01-01

    Introduction Nitration of tyrosine and tyrosine-containing proteins and their roles in pathophysiology have just recently been reviewed. Despite low yields of tyrosine modifications, nitration of tyrosine residues may inactivate important proteins. Nitrotyrosine can be formed by various nitrating agents, including peroxynitrite. Thus, the occurrence of nitrotyrosine-containing proteins in vivo should be regarded as a general indication of tissue damage induced by reactive nitrogen species such as peroxynitrite. This strongly suggests that peroxynitrite could be formed in vivo under certain pathophysiological conditions. Objective Our aim in this study was to elucidate the effect of cigarette smoke (CS) on nitrotyrosine formation in saliva proteins. Methods We exposed saliva to CS, in vitro, and used Western Blotting (WB) and monoclonal anti-nitrotyrosine antibody to assess the level of saliva protein nitration. Results As saliva contains extensive amounts of nitrites, it was no surprise that at basal levels, saliva proteins, albumin, and ?-amylase all were already nitrated. The WB also revealed that with continuous exposure to CS the tyrosine nitration of both albumin and ?-amylase is declining significantly after 3 h. A quite similar effect was seen after exposure to aldehydes, but to a less extent as compared to CS. Exposure of nitrotyrosine-modified bovine serum albumin (BSA-N) to aldehydes, produced a similar effect, meaning a decrease in tyrosine nitration. Conclusions These findings might be explained by the possible ability of CS aldehydes to reduce protein-bound nitro group to an amine. Another proposed mechanism is that CS unsaturated aldehydes react with proteins mainly through Michael addition reaction; leading to the generation of stable aldehyde-protein adducts (APA). Thus, it may react with nitro groups of saliva proteins, like albumin or ?-amylase, to generate APA, which ultimately, may not be recognized by our antibody. Another possible mechanism, is interaction between the aldehyde group with the hydroxyl group of the 3-nitrotyrosine, forming a hemiacetal, which is not recognized by the antibody. This mechanism might explain the difference in the 'denitration' effects caused by the saturated aldehyde acetaldehyde, which exists in large amounts in CS, and unsaturated aldehydes. Therefore, it is possible that the main player in the CS smoke "denitration" effect on salivary proteins is the aldehyde group and not the double bond of unsaturated aldehydes. PMID:21147654

  4. Measuring salivary analytes from free-ranging monkeys.

    PubMed

    Higham, James P; Vitale, Alison B; Rivera, Adaris Mas; Ayala, James E; Maestripieri, Dario

    2010-12-01

    Studies of large free-ranging mammals have been revolutionized by non-invasive methods for assessing physiology, which usually involve the measurement of fecal or urinary biomarkers. However, such techniques are limited by numerous factors. To expand the range of physiological variables measurable non-invasively from free-ranging primates, we developed techniques for sampling monkey saliva by offering monkeys ropes with oral swabs sewn on the ends. We evaluated different attractants for encouraging individuals to offer samples, and proportions of individuals in different age/sex categories willing to give samples. We tested the saliva samples we obtained in three commercially available assays: cortisol, salivary alpha amylase, and secretory immunoglobulin A. We show that habituated free-ranging rhesus macaques will give saliva samples voluntarily without training, with 100% of infants, and over 50% of adults willing to chew on collection devices. Our field methods are robust even for analytes that show poor recovery from cotton, and/or that have concentrations dependent on salivary flow rate. We validated the cortisol and SAA assays for use in rhesus macaques by showing aspects of analytical validation, such as that samples dilute linearly and in parallel to assay standards. We also found that values measured correlated with biologically meaningful characteristics of sampled individuals (age and dominance rank). The SIgA assay tested did not react to samples. Given the wide range of analytes measurable in saliva but not in feces or urine, our methods considerably improve our ability to study physiological aspects of the behavior and ecology of free-ranging primates, and are also potentially adaptable to other mammalian taxa. PMID:20837036

  5. Modulation of Sodium/Iodide Symporter Expression in the Salivary Gland

    PubMed Central

    La Perle, Krista M.D.; Kim, Dong Chul; Hall, Nathan C.; Bobbey, Adam; Shen, Daniel H.; Nagy, Rebecca S.; Wakely, Paul E.; Lehman, Amy; Jarjoura, David

    2013-01-01

    Background Physiologic iodide-uptake, mediated by the sodium/iodide symporter (NIS), in the salivary gland confers its susceptibility to radioactive iodine–induced damage following 131I treatment of thyroid cancer. Subsequent quality of life for thyroid cancer survivors can be decreased due to recurrent sialoadenitis and persistent xerostomia. NIS expression at the three principal salivary duct components in various pathological conditions was examined to better our understanding of NIS modulation in the salivary gland. Methods NIS expression was evaluated by immunohistochemistry in human salivary gland tissue microarrays constructed of normal, inflamed, and neoplastic salivary tissue cores. Cumulative 123I radioactivity reflecting the combination of NIS activity with clearance of saliva secretion in submandibular and parotid salivary glands was evaluated by single-photon emission computed tomography/computed tomography imaging 24 hours after 123I administration in 50 thyroid cancer patients. Results NIS is highly expressed in the basolateral membranes of the majority of striated ducts, yet weakly expressed in few intercalated and excretory duct cells. The ratio of 123I accumulation between parotid and submandibular glands is 2.38±0.19. However, the corresponding ratio of 123I accumulation normalized by volume of interest is 1.19±0.06. The percentage of NIS-positive striated duct cells in submandibular salivary glands was statistically greater than in parotid salivary glands, suggesting a higher clearance rate of saliva secretion in submandibular salivary glands. NIS expression in striated ducts was heterogeneously decreased or absent in sialoadenitis. Most ductal salivary gland tumors did not express NIS. However, Warthin's tumors of striated duct origin exhibited consistent and intense NIS staining, corresponding with radioactive iodine uptake. Conclusions NIS expression is tightly modulated during the transition of intercalated to striated ducts and striated to excretory ducts in salivary ductal cells. NIS expression in salivary glands is decreased during inflammation and tumor formation. Further investigation may identify molecular targets and/or pharmacologic agents that allow selective inhibition of NIS expression/activity in salivary glands during radioactive iodine treatment. PMID:23441638

  6. Organic solvent tolerance of halophilic alpha-amylase from a Haloarchaeon, Haloarcula sp. strain S-1.

    PubMed

    Fukushima, Tadamasa; Mizuki, Toru; Echigo, Akinobu; Inoue, Akira; Usami, Ron

    2005-02-01

    A halophilic archaeon, Haloarcula sp. strain S-1, produced extracellular organic solvent-tolerant alpha-amylase. Molecular mass of the enzyme was estimated to be 70 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This amylase exhibited maximal activity at 50 degrees C in buffer containing 4.3 M NaCl, pH 7.0. Moreover, the enzyme was active and stable in various organic solvents (benzene, toluene, and chloroform, etc.). Activity was not detected at low ionic strengths, but it was detected in the presence of chloroform at low salt concentrations. On the other hand, no activity was detected in the presence of ethyl alcohol and acetone. PMID:15378403

  7. Enhanced Production and Characterization of a Solvent Stable Amylase from Solvent Tolerant Bacillus tequilensis RG-01: Thermostable and Surfactant Resistant

    PubMed Central

    Tiwari, Soni; Shukla, Neha; Mishra, Pooja; Gaur, Rajeeva

    2014-01-01

    Ten bacterial strains isolated from the soil samples in the presence of cyclohexane were screened for amylase production. Among them, culture RG-01 was adjudged as the best amylase producer and was identified as Bacillus tequilensis from MTCC, Chandigarh. The isolate showed maximum amylase production (8100?U/mL) in the presence of starch, peptone, and Ca2+ ions at 55°C pH 7.0 within 24?h of incubation. The enzyme was stable in the presence of n-dodecane, isooctane, n-decane, xylene, toluene, n-hexane, n-butanol, and cyclohexane, respectively. The presence of benzene, methanol, and ethanol marginally reduced the amylase stability, respectively. The enzyme was showed it 100% activity at 55°C and pH 7.0 with 119% and 127% stability at 55°C and pH 7.0, respectively. The enzyme was also stable in the presence of SDS, Tween-40, Tween-60, and Tween-80 (1%) and was found stimulatory effect, respectively. Only Triton-X-100 showed a moderate inhibitory effect (5%) on amylase activity. This isolate (Bacillus tequilensis RG-01) may be useful in several industrial applications owing to its thermotolerant and organic solvents and surfactants resistance characteristics. PMID:25401163

  8. Comprehensive enzymatic analysis of the amylolytic system in the digestive fluid of the sea hare, Aplysia kurodai: Unique properties of two ?-amylases and two ?-glucosidases

    PubMed Central

    Tsuji, Akihiko; Nishiyama, Nami; Ohshima, Miki; Maniwa, Saori; Kuwamura, Shuji; Shiraishi, Masataka; Yuasa, Keizo

    2014-01-01

    Sea lettuce (Ulva pertusa) is a nuisance species of green algae that is found all over the world. East-Asian species of the marine gastropod, the sea hare Aplysia kurodai, shows a clear feeding preference for sea lettuce. Compared with cellulose, sea lettuce contains a higher amount of starch as a storage polysaccharide. However, the entire amylolytic system in the digestive fluid of A. kurodai has not been studied in detail. We purified ?-amylases and ?-glucosidases from the digestive fluid of A. kurodai and investigated the synergistic action of these enzymes on sea lettuce. A. kurodai contain two ?-amylases (59 and 80 kDa) and two ?-glucosidases (74 and 86 kDa). The 59-kDa ?-amylase, but not the 80-kDa ?-amylase, was markedly activated by Ca2+ or Cl?. Both ?-amylases degraded starch and maltoheptaose, producing maltotriose, maltose, and glucose. Glucose production from starch was higher with 80-kDa ?-amylase than with 59-kDa ?-amylase. Kinetic analysis indicated that 74-kDa ?-glucosidase prefers short ?-1,4-linked oligosaccharide, whereas 86-kDa ?-glucosidase prefers large ?-1,6 and ?-1,4-linked polysaccharides such as glycogen. When sea lettuce was used as a substrate, a 2-fold greater amount of glucose was released by treatment with 59-kDa ?-amylase and 74-kDa ?-glucosidase than by treatment with 45-kDa cellulase and 210-kDa ?-glucosidase of A. kurodai. Unlike mammals, sea hares efficiently digest sea lettuce to glucose by a combination of two ?-amylases and two ?-glucosidases in the digestive fluids without membrane-bound maltase–glucoamylase and sucrase–isomaltase complexes. PMID:25161866

  9. Comprehensive enzymatic analysis of the amylolytic system in the digestive fluid of the sea hare, Aplysia kurodai: Unique properties of two ?-amylases and two ?-glucosidases.

    PubMed

    Tsuji, Akihiko; Nishiyama, Nami; Ohshima, Miki; Maniwa, Saori; Kuwamura, Shuji; Shiraishi, Masataka; Yuasa, Keizo

    2014-01-01

    Sea lettuce (Ulva pertusa) is a nuisance species of green algae that is found all over the world. East-Asian species of the marine gastropod, the sea hare Aplysia kurodai, shows a clear feeding preference for sea lettuce. Compared with cellulose, sea lettuce contains a higher amount of starch as a storage polysaccharide. However, the entire amylolytic system in the digestive fluid of A. kurodai has not been studied in detail. We purified ?-amylases and ?-glucosidases from the digestive fluid of A. kurodai and investigated the synergistic action of these enzymes on sea lettuce. A. kurodai contain two ?-amylases (59 and 80 kDa) and two ?-glucosidases (74 and 86 kDa). The 59-kDa ?-amylase, but not the 80-kDa ?-amylase, was markedly activated by Ca(2+) or Cl(-). Both ?-amylases degraded starch and maltoheptaose, producing maltotriose, maltose, and glucose. Glucose production from starch was higher with 80-kDa ?-amylase than with 59-kDa ?-amylase. Kinetic analysis indicated that 74-kDa ?-glucosidase prefers short ?-1,4-linked oligosaccharide, whereas 86-kDa ?-glucosidase prefers large ?-1,6 and ?-1,4-linked polysaccharides such as glycogen. When sea lettuce was used as a substrate, a 2-fold greater amount of glucose was released by treatment with 59-kDa ?-amylase and 74-kDa ?-glucosidase than by treatment with 45-kDa cellulase and 210-kDa ?-glucosidase of A. kurodai. Unlike mammals, sea hares efficiently digest sea lettuce to glucose by a combination of two ?-amylases and two ?-glucosidases in the digestive fluids without membrane-bound maltase-glucoamylase and sucrase-isomaltase complexes. PMID:25161866

  10. Molecular cloning and characterization of an ?-amylase cDNA highly expressed in major feeding stages of the coffee berry borer, Hypothenemus hampei.

    PubMed

    Bezerra, C A; Macedo, L L P; Amorim, T M L; Santos, V O; Fragoso, R R; Lucena, W A; Meneguim, A M; Valencia-Jimenez, A; Engler, G; Silva, M C M; Albuquerque, E V S; Grossi-de-Sa, M F

    2014-12-10

    ?-Amylases are common enzymes responsible for hydrolyzing starch. Insect-pests, whose larvae develop in seeds, rely obligatorily on ?-amylase activity to digest starch, as their major food source. Considering the relevance of insect ?-amylases and the natural ?-amylase inhibitors present in seeds to protect from insect damage, we report here the molecular cloning and nucleotide sequence of the full-length AmyHha cDNA of the coffee berry borer, Hypothenemus hampei, a major insect-pest of coffee crops. The AmyHha sequence has 1879 bp, containing a 1458 bp open reading frame, which encodes a predicted protein with 485 amino acid residues, with a predicted molecular mass of 51.2 kDa. The deduced protein showed 55-79% identity to other insect ?-amylases, including Anthonomus grandis, Ips typographus and Sitophilus oryzae ?-amylases. In depth analysis revealed that the highly conserved three amino acid residues (Asp184, Glu220, and Asp285), which compose the catalytic site are also presented in AmyHha amylase. The AmyHha gene seems to be a single copy in the haploid genome and AmyHha transcription levels were found higher in L2 larvae and adult insects, both corresponding to major feeding phases. Modeling of the AmyHha predicted protein uncovered striking structural similarities to the Tenebrio molitor ?-amylase also displaying the same amino acid residues involved in enzyme catalysis (Asp184, Glu220 and Asp285). Since AmyHha gene was mostly transcribed in the intestinal tract of H. hampei larvae, the cognate ?-amylase could be considered a high valuable target to coffee bean insect control by biotechnological strategies. PMID:25264343

  11. Anatomy, biogenesis and regeneration of salivary glands.

    PubMed

    Holmberg, Kyle V; Hoffman, Matthew P

    2014-01-01

    An overview of the anatomy and biogenesis of salivary glands is important in order to understand the physiology, functions and disorders associated with saliva. A major disorder of salivary glands is salivary hypofunction and resulting xerostomia, or dry mouth, which affects hundreds of thousands of patients each year who suffer from salivary gland diseases or undergo head and neck cancer treatment. There is currently no curative therapy for these patients. To improve these patients' quality of life, new therapies are being developed based on findings in salivary gland cell and developmental biology. Here we discuss the anatomy and biogenesis of the major human salivary glands and the rodent submandibular gland, which has been used extensively as a research model. We also include a review of recent research on the identification and function of stem cells in salivary glands, and the emerging field of research suggesting that nerves play an instructive role during development and may be essential for adult gland repair and regeneration. Understanding the molecular mechanisms involved in gland biogenesis provides a template for regenerating, repairing or reengineering diseased or damaged adult human salivary glands. We provide an overview of 3 general approaches currently being developed to regenerate damaged salivary tissue, including gene therapy, stem cell-based therapy and tissue engineering. In the future, it may be that a combination of all three will be used to repair, regenerate and reengineer functional salivary glands in patients to increase the secretion of their saliva, the focus of this monograph. PMID:24862590

  12. Single-step purification of a recombinant thermostable alpha-amylase after solubilization of the enzyme from insoluble aggregates.

    PubMed

    Linden, A; Niehaus, F; Antranikian, G

    2000-01-14

    The expression of the gene encoding a thermostable alpha-amylase (EC 3.2.1.1) (optimal activity at 100 degrees C) from the hyperthermophilic archaeon Pyrococcus woesei in the mesophilic hosts Escherichia coli and Halomonas elongata resulted in the formation of insoluble aggregates. More than 85% of the recombinant enzyme was present within the cells as insoluble but catalytically active aggregates. The recombinant alpha-amylase was purified to homogeneity in a single step by hydrophobic interaction chromatography on a phenyl superose column after solubilization of the enzyme under nondenaturing conditions. The enzyme was purified 258-fold with a final yield of 54%. PMID:10681062

  13. Facial Vibrotactile Stimulation Activates the Parasympathetic Nervous System: Study of Salivary Secretion, Heart Rate, Pupillary Reflex, and Functional Near-Infrared Spectroscopy Activity

    PubMed Central

    Hiraba, Hisao; Inoue, Motoharu; Gora, Kanako; Sato, Takako; Nishimura, Satoshi; Yamaoka, Masaru; Kumakura, Ayano; Ono, Shinya; Wakasa, Hirotugu; Nakayama, Enri; Abe, Kimiko; Ueda, Koichiro

    2014-01-01

    We previously found that the greatest salivation response in healthy human subjects is produced by facial vibrotactile stimulation of 89?Hz frequency with 1.9??m amplitude (89?Hz-S), as reported by Hiraba et al. (2012, 20011, and 2008). We assessed relationships between the blood flow to brain via functional near-infrared spectroscopy (fNIRS) in the frontal cortex and autonomic parameters. We used the heart rate (HRV: heart rate variability analysis in RR intervals), pupil reflex, and salivation as parameters, but the interrelation between each parameter and fNIRS measures remains unknown. We were to investigate the relationship in response to established paradigms using simultaneously each parameter-fNIRS recording in healthy human subjects. Analysis of fNIRS was examined by a comparison of various values between before and after various stimuli (89?Hz-S, 114?Hz-S, listen to classic music, and “Ahh” vocalization). We confirmed that vibrotactile stimulation (89?Hz) of the parotid glands led to the greatest salivation, greatest increase in heart rate variability, and the most constricted pupils. Furthermore, there were almost no detectable differences between fNIRS during 89?Hz-S and fNIRS during listening to classical music of fans. Thus, vibrotactile stimulation of 89?Hz seems to evoke parasympathetic activity. PMID:24511550

  14. Facial vibrotactile stimulation activates the parasympathetic nervous system: study of salivary secretion, heart rate, pupillary reflex, and functional near-infrared spectroscopy activity.

    PubMed

    Hiraba, Hisao; Inoue, Motoharu; Gora, Kanako; Sato, Takako; Nishimura, Satoshi; Yamaoka, Masaru; Kumakura, Ayano; Ono, Shinya; Wakasa, Hirotugu; Nakayama, Enri; Abe, Kimiko; Ueda, Koichiro

    2014-01-01

    We previously found that the greatest salivation response in healthy human subjects is produced by facial vibrotactile stimulation of 89 Hz frequency with 1.9 ? m amplitude (89 Hz-S), as reported by Hiraba et al. (2012, 20011, and 2008). We assessed relationships between the blood flow to brain via functional near-infrared spectroscopy (fNIRS) in the frontal cortex and autonomic parameters. We used the heart rate (HRV: heart rate variability analysis in RR intervals), pupil reflex, and salivation as parameters, but the interrelation between each parameter and fNIRS measures remains unknown. We were to investigate the relationship in response to established paradigms using simultaneously each parameter-fNIRS recording in healthy human subjects. Analysis of fNIRS was examined by a comparison of various values between before and after various stimuli (89 Hz-S, 114 Hz-S, listen to classic music, and "Ahh" vocalization). We confirmed that vibrotactile stimulation (89 Hz) of the parotid glands led to the greatest salivation, greatest increase in heart rate variability, and the most constricted pupils. Furthermore, there were almost no detectable differences between fNIRS during 89 Hz-S and fNIRS during listening to classical music of fans. Thus, vibrotactile stimulation of 89 Hz seems to evoke parasympathetic activity. PMID:24511550

  15. Cloning and expression of a Bacillus coagulans amylase gene in Escherichia coli

    Microsoft Academic Search

    Pierre Cornelis; Colette Digneffe; Karine Willemot

    1982-01-01

    A partial EcoRI fragment of Bacillus coagulans DNA cloned in an Escherichia coli K12 bacteriophage ? host-vector system was shown to direct the synthesis of a thermostable a-amylase whose activity could be detected in situ on petri plates using the iodine staining method. A 3.31 kb EcoRI fragment containing the active gene with its own promoter was subcloned in pBR322;

  16. Inter organ comparison of amylases and starch content in mungbean seedlings

    Microsoft Academic Search

    Narinder Kaur; Prabhdeep Kaur; Anil K. Gupta

    2001-01-01

    During seedling growth of mungbean in dark, depletion of cotyledonary starch is reflected by an increase in starch content\\u000a of root and shoot. With progress of seedling growth, amylolytic activity increases in all organs i.e. cotyledons, shoots and roots. A rapid turnover of starch in shoots and roots has been proposed. Amylase activity of seedlings\\u000a was in the order of

  17. Halophilic characterization of starch-binding domain from Kocuria varians ?-amylase

    Microsoft Academic Search

    Rui Yamaguchi; Yasuhiro Inoue; Hiroko Tokunaga; Matsujiro Ishibashi; Tsutomu Arakawa; Jun-ichi Sumitani; Takashi Kawaguchi; Masao Tokunaga

    The tandem starch-binding domains (KvSBD) located at carboxy-terminal region of halophilic ?-amylase from moderate halophile, Kocuria varians, were expressed in E. coli with amino-terminal hexa-His-tag and purified to homogeneity. The recombinant KvSBD showed binding activity to raw starch granules at low to high salt concentrations. The binding activity of KvSBD to starch was fully reversible after heat-treatment at 85°C. Circular

  18. Tactics for modeling multiple salivary analyte data in relation to behavior problems: Additive, ratio, and interaction effects.

    PubMed

    Chen, Frances R; Raine, Adrian; Granger, Douglas A

    2015-01-01

    Individual differences in the psychobiology of the stress response have been linked to behavior problems in youth yet most research has focused on single signaling molecules released by either the hypothalamic-pituitary-adrenal axis or the autonomic nervous system. As our understanding about biobehavioral relationships develops it is clear that multiple signals from the biological stress systems work in coordination to affect behavior problems. Questions are raised as to whether coordinated effects should be statistically represented as ratio or interactive terms. We address this knowledge gap by providing a theoretical overview of the concepts and rationales, and illustrating the analytical tactics. Salivary samples collected from 446 youth aged 11-12 were assayed for salivary alpha-amylase (sAA), dehydroepiandrosterone-sulfate (DHEA-s) and cortisol. Coordinated effect of DHEA-s and cortisol, and coordinated effect of sAA and cortisol on externalizing and internalizing problems (Child Behavior Checklist) were tested with the ratio and the interaction approaches using multi-group path analysis. Findings consistent with previous studies include a positive association between cortisol/DHEA-s ratio and internalizing problems; and a negative association between cortisol and externalizing problems conditional on low levels of sAA. This study highlights the importance of matching analytical strategy with research hypothesis when integrating salivary bioscience into research in behavior problems. Recommendations are made for investigating multiple salivary analytes in relation to behavior problems. PMID:25462892

  19. Two Strategies for Microbial Production of an Industrial Enzyme-Alpha-Amylase

    NASA Technical Reports Server (NTRS)

    Bernhardsdotter, Eva C. M. J.; Garriott, Owen; Pusey, Marc L.; Ng, Joseph D.

    2003-01-01

    Extremophiles are microorganisms that thrive in, from an anthropocentric view, extreme environments including hot springs, soda lakes and arctic water. This ability of survival at extreme conditions has rendered extremophiles to be of interest in astrobiology, evolutionary biology as well as in industrial applications. Of particular interest to the biotechnology industry are the biological catalysts of the extremophiles, the extremozymes, whose unique stabilities at extreme conditions make them potential sources of novel enzymes in industrial applications. There are two major approaches to microbial enzyme production. This entails enzyme isolation directly from the natural host or creating a recombinant expression system whereby the targeted enzyme can be overexpressed in a mesophilic host. We are employing both methods in the effort to produce alpha-amylases from a hyperthermophilic archaeon (Thermococcus) isolated from a hydrothermal vent in the Atlantic Ocean, as well as from alkaliphilic bacteria (Bacillus) isolated from a soda lake in Tanzania. Alpha-amylases catalyze the hydrolysis of internal alpha-1,4-glycosidic linkages in starch to produce smaller sugars. Thermostable alpha-amylases are used in the liquefaction of starch for production of fructose and glucose syrups, whereas alpha-amylases stable at high pH have potential as detergent additives. The alpha-amylase encoding gene from Thermococcus was PCR amplified using carefully designed primers and analyzed using bioinformatics tools such as BLAST and Multiple Sequence Alignment for cloning and expression in E.coli. Four strains of Bacillus were grown in alkaline starch-enriched medium of which the culture supernatant was used as enzyme source. Amylolytic activity was detected using the starch-iodine method.

  20. Development of an amylolytic Lactobacillus plantarum silage strain expressing the Lactobacillus amylovorus alpha-amylase gene.

    PubMed Central

    Fitzsimons, A; Hols, P; Jore, J; Leer, R J; O'Connell, M; Delcour, J

    1994-01-01

    An amylolytic Lactobacillus plantarum silage strain with the starch-degrading ability displayed by Lactobacillus amylovorus was developed. An active fragment of the gene coding for alpha-amylase production in L. amylovorus was cloned and integrated into the chromosome of the competitive inoculant strain L. plantarum Lp80 at the cbh locus. The alpha-amylase gene fragment was also introduced into L. plantarum Lp80 on an autoreplicative plasmid. Both constructions were also performed in the laboratory strain L. plantarum NCIB8826. All four recombinant strains secreted levels of amylase ranging from 23 to 69 U/liter, compared with 47 U/liter for L. amylovorus. Secretion levels were higher in L. plantarum NCIB8826 than in L. plantarum Lp80 derivatives and were higher in recombinant strains containing autoreplicative plasmids than in the corresponding integrants. The L. plantarum Lp80 derivative containing the L. amylovorus alpha-amylase gene fragment integrated into the host chromosome secreted alpha-amylase to a level comparable to that of L. amylovorus and was stable over 50 generations of growth under nonselective conditions. It grew to a higher cell density than either the parent strain or L. amylovorus in MRS medium containing a mixture of starch and glucose as the fermentable carbohydrate source. This recombinant alpha-amylolytic L. plantarum strain would therefore seem to have considerable potential as a silage inoculant for crops such as alfalfa, in which water-soluble carbohydrate levels are frequently low but starch is present as an alternative carbohydrate source. Images PMID:7986030

  1. Development of an amylolytic Lactobacillus plantarum silage strain expressing the Lactobacillus amylovorus alpha-amylase gene.

    PubMed

    Fitzsimons, A; Hols, P; Jore, J; Leer, R J; O'Connell, M; Delcour, J

    1994-10-01

    An amylolytic Lactobacillus plantarum silage strain with the starch-degrading ability displayed by Lactobacillus amylovorus was developed. An active fragment of the gene coding for alpha-amylase production in L. amylovorus was cloned and integrated into the chromosome of the competitive inoculant strain L. plantarum Lp80 at the cbh locus. The alpha-amylase gene fragment was also introduced into L. plantarum Lp80 on an autoreplicative plasmid. Both constructions were also performed in the laboratory strain L. plantarum NCIB8826. All four recombinant strains secreted levels of amylase ranging from 23 to 69 U/liter, compared with 47 U/liter for L. amylovorus. Secretion levels were higher in L. plantarum NCIB8826 than in L. plantarum Lp80 derivatives and were higher in recombinant strains containing autoreplicative plasmids than in the corresponding integrants. The L. plantarum Lp80 derivative containing the L. amylovorus alpha-amylase gene fragment integrated into the host chromosome secreted alpha-amylase to a level comparable to that of L. amylovorus and was stable over 50 generations of growth under nonselective conditions. It grew to a higher cell density than either the parent strain or L. amylovorus in MRS medium containing a mixture of starch and glucose as the fermentable carbohydrate source. This recombinant alpha-amylolytic L. plantarum strain would therefore seem to have considerable potential as a silage inoculant for crops such as alfalfa, in which water-soluble carbohydrate levels are frequently low but starch is present as an alternative carbohydrate source. PMID:7986030

  2. Optimised amylases extraction from oat seeds and its impact on bread properties.

    PubMed

    Ben Halima, Nihed; Borchani, Maha; Fendri, Imen; Khemakhem, Bassem; Gosset, David; Baril, Patrick; Pichon, Chantal; Ayadi, Mohamed-Ali; Abdelkafi, Slim

    2015-01-01

    Statistical approaches were employed for the optimisation of the extraction of amylolytic activity from oat (Avena sativa) seeds. The application of the response surface methodology allows us to determine a set of optimal conditions (ratio seed weight/buffer volume 0.1, germination days 10 days, temperature 20 °C and pH 5.6). Experiments carried out under these conditions led to amylase production yield of 91 U/g. Its maximal activity was in the pH 5.6 and at 55 °C. Study of the incorporation of the optimised oat extract into the bread formulation revealed an improvement of the sensory quality and the textural properties of fresh and stored bread. Three-dimensional elaborations of Confocal Laser Scanning Microscopy (CLSM) images were performed on crumb of the different breads to evaluate the influence of amylase activity on microstructure. The result showed improved baking characteristics as well as overall microscopic and macroscopic appearance. PMID:25453287

  3. Enzyme Assay Guided Isolation of an ?-Amylase Inhibitor Flavonoid from Vaccinium arctostaphylos Leaves

    PubMed Central

    Nickavar, Bahman; Amin, Gholamreza

    2011-01-01

    The management of postprandial hyperglycemia is an important strategy in the control of diabetes mellitus and complications associated with the disease, especially in the diabetes type 2. Therefore, inhibitors of carbohydrate hydrolyzing enzymes can be useful in the treatment of diabetes and medicinal plants can offer an attractive strategy for the purpose. Vaccinium arctostaphylos leaves are considered useful for the treatment of diabetes mellitus in some countries. In our research for antidiabetic compounds from natural sources, we found that the methanol extract of the leaves of V. arctostaphylos displayed a potent inhibitory activity on pancreatic ?-amylase activity (IC50 = 0.53 (0.53 – 0.54) mg/mL). The bioassay-guided fractionation of the extract resulted in the isolation of quercetin as an active ?-amylase inhibitor. Quercetin showed a dose-dependent inhibitory effect with IC50 value 0.17 (0.16 – 0.17) mM. PMID:24250422

  4. Spectral effect: each population must have its own normal midnight salivary cortisol reference values determined

    PubMed Central

    Tanakol, Refik; Karpuzoglu, Hande; Abbasoglu, Semra; Yarman, Sema; Boztepe, Harika; Alagol, Faruk

    2013-01-01

    Introduction The mesurement of midnight salivary cortisol provides the most sensitive method for screening of Cushing's sendrome. However the clinical significance of spectral error is the requirement for determination of normal reference values in each population for each test, which will be used as the diagnostic method. Salivary cortisol levels may be affected by individual factors such as nutrition, sleep, medication, activity, and gender. Being a non-invasive method, midnight salivary cortisol (MSC) has been used as a valuable indicator of free plasma cortisol. Material and methods Midnight salivary cortisol was assessed in randomly selected 100 Turkish patents who underwent to a detailed physical examination. Saliva samples were collected at 00:00 to plastic tubes with the help of plastic pipettes, without brushing their teeth, but after rinsing their mouth. Salivary cortisol was measured with luminescense immunoassay kit. Differences and correlations were analysed. Results The mean midnight salivary cortisol of the healthy population was 0.21 ±0.03 µg/dl. Body mass index, age, sex, smoking, exercise, educational status alcohol, had no effect on the MSC. Conclusions Consequently, normal salivary cortisol reference ranges must be used for different assays and different populations in order to evaluate more accurately pituitary-adrenal axis pathology in clinical practice. PMID:24273572

  5. Direct radioimmunoassay (RIA) of salivary testosterone: correlation with free and total serium testosterone

    SciTech Connect

    Vittek, J.; L'Hommedieu, D.G.; Gordon, G.G.; Rappaport, S.C.; Southren, A.L.

    1985-08-26

    Simple and sensitive direct RIA for determination of salivary testosterone was developed by using RSL NOSOLVEX TM (125 1) kit produced by Radioassay System Laboratories (Carcon, California). In addition, a relationship between salivary and serum free and total testosterone concentrations was studied in randomly selected 45 healthy subjects, 5 females on oral contraceptive pills and 28 hypertensive patients on various treatment regimens. The lowest weight of testosterone detectable by the modified method was equivalent to 1 pg/ml of saliva, taking into account analytical variability. Intra- and interassay coefficients of variation were 5.09 +/- 2.7% and 8.2 +/- 5.9% respectively. Statistically significant correlations were found between salivary and serum free testosterone (r = 0.97) and salivary and serum total testosterone concentrations (r = 0.70 - 0.87). The exception to this was a group of hypertensive females in which no correlation (r = 0.14) between salivary and total serum testosterone was found. It is also of interest that, while salivary testosterone was significantly increased in subjects taking oral contraceptives and most of the hypertensive patients, the total serum testosterone concentration was in normal range. These findings suggest that the determination of salivary testosterone is a reliable method to detect changes in the concentration of available biologically active hormone in the circulation. 21 references, 4 figures, 1 table.

  6. What Are the Key Statistics about Salivary Gland Cancer?

    MedlinePLUS

    ... for salivary gland cancer? What are the key statistics about salivary gland cancer? Salivary gland cancers are ... be better or worse than this.) For more statistics related to survival, see the section “ Survival rates ...

  7. Comparisons of salivary proteins from five aphid (Hemiptera: Aphididae) species.

    PubMed

    Cooper, W Rodney; Dillwith, Jack W; Puterka, Gary J

    2011-02-01

    Aphid (Hemiptera: Aphididae) saliva, when injected into host plants during feeding, causes physiological changes in hosts that facilitate aphid feeding and cause injury to plants. Comparing salivary constituents among aphid species could help identify which salivary products are universally important for general aphid feeding processes, which products are involved with specific host associations, or which products elicit visible injury to hosts. We compared the salivary proteins from five aphid species, namely, Diuraphis noxia (Kurdjumov), D. tritici (Gillette), D. mexicana (Baker), Schizaphis graminum (Rondani), and Acyrthosiphon pisum (Harris). A 132-kDa protein band was detected from the saliva of all five species using sodium dodecyl sulfate polyacrylamide gel electrophoresis. Alkaline phosphatase activity was detected from the saliva of all five species and may have a universal role in the feeding process of aphids. The Diuraphis species cause similar visible injury to grass hosts, and nine electrophoretic bands were unique to the saliva of these three species. S. graminum shares mutual hosts with the Diuraphis species, but visible injury to hosts caused by S. graminum feeding differs from that of Diuraphis feeding. Only two mutual electrophoretic bands were visualized in the saliva of Diuraphis and S. graminum. Ten unique products were detected from the saliva of A. pisum, which feeds on dicotyledonous hosts. Our comparisons of aphid salivary proteins revealed similarities among species which cause similar injury on mutual hosts, fewer similarities among species that cause different injury on mutual hosts, and little similarity among species which feed on unrelated hosts. PMID:22182624

  8. Optimization of Amylase Application in Raw Sugar Manufacture. Part I: Characterization of Commercial Alpha-Amylases

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In recent years there have been warnings by some U.S. refineries that there may be a penalty for high starch concentrations in raw sugar if starch control is not improved. Most commercial amylases used by the U.S. sugar industry to control starch have intermediate temperature stability (up to 85 de...

  9. Method for using a yeast alpha-amylase promoter

    DOEpatents

    Gao, Johnway (Richland, WA); Skeen, Rodney S. (Pendleton, OR); Hooker, Brian S. (Kennewick, WA); Anderson, Daniel B. (Pasco, WA)

    2003-04-22

    The present invention provides the promoter clone discovery of an alpha-amylase gene of a starch utilizing yeast strain Schwanniomyces castellii. The isolated alpha-amylase promoter is an inducible promoter, which can regulate strong gene expression in starch culture medium.

  10. Application of microbial ?-amylase in industry – A review

    PubMed Central

    de Souza, Paula Monteiro; de Oliveira Magalhães, Pérola

    2010-01-01

    Amylases are one of the main enzymes used in industry. Such enzymes hydrolyze the starch molecules into polymers composed of glucose units. Amylases have potential application in a wide number of industrial processes such as food, fermentation and pharmaceutical industries. ?-Amylases can be obtained from plants, animals and microorganisms. However, enzymes from fungal and bacterial sources have dominated applications in industrial sectors. The production of ?-amylase is essential for conversion of starches into oligosaccharides. Starch is an important constituent of the human diet and is a major storage product of many economically important crops such as wheat, rice, maize, tapioca, and potato. Starch-converting enzymes are used in the production of maltodextrin, modified starches, or glucose and fructose syrups. A large number of microbial ?-amylases has applications in different industrial sectors such as food, textile, paper and detergent industries. The production of ?-amylases has generally been carried out using submerged fermentation, but solid state fermentation systems appear as a promising technology. The properties of each ?-amylase such as thermostability, pH profile, pH stability, and Ca-independency are important in the development of fermentation process. This review focuses on the production of bacterial and fungal ?-amylases, their distribution, structural-functional aspects, physical and chemical parameters, and the use of these enzymes in industrial applications. PMID:24031565

  11. Amlioration des plantes Polymorphisme de l'? amylase chez le pcher.

    E-print Network

    Paris-Sud XI, Université de

    , the loci which determined the type of flower and the color of the flesh are also independent (table V / groupe de liaison Summary — ?-Amylase polymorphism in peach: a genetic study. &alpha the locus of ?-amylase and that which determines the type of flower (fig 2; table III), the locus

  12. The Amylase Project: Creating a Classroom of Biotechnologists.

    ERIC Educational Resources Information Center

    Sweeney, Diane

    1998-01-01

    A biotechnologist-turned-teacher introduces a series of laboratory modules incorporating concepts from microbiology, cellular biology, molecular biology, biochemistry, and evolution. The Amylase Project aims to distill the biotechnology process into a few short steps using amylase, the easiest enzyme to detect of those commonly produced by…

  13. Influence of substratum hydrophobicity on salivary pellicles: organization or composition?

    PubMed

    Aroonsang, Watcharapong; Sotres, Javier; El-Schich, Zahra; Arnebrant, Thomas; Lindh, Liselott

    2014-10-01

    Different physico-chemical properties (eg adsorption kinetics, thickness, viscoelasticity, and mechanical stability) of adsorbed salivary pellicles depend on different factors, including the properties (eg charge, roughness, wettability, and surface chemistry) of the substratum. Whether these differences in the physico-chemical properties are a result of differences in the composition or in the organization of the pellicles is not known. In this work, the influence of substratum wettability on the composition of the pellicle was studied. For this purpose, pellicles eluted from substrata of different but well-characterized wettabilities were examined by means of sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that substratum hydrophobicity did not have a major impact on pellicle composition. In all substrata, the major pellicle components were found to be cystatins, amylases and large glycoproteins, presumably mucins. In turn, interpretation of previously reported data based on the present results suggests that variations in substratum wettability mostly affect the organization of the pellicle components. PMID:25377485

  14. Review of Salivary Gland Neoplasms

    PubMed Central

    To, Victor Shing Howe; Chan, Jimmy Yu Wai; Tsang, Raymond K. Y.; Wei, William I.

    2012-01-01

    Salivary gland tumours most often present as painless enlarging masses. Most are located in the parotid glands and most are benign. The principal hurdle in their management lies in the difficulty in distinguishing benign from malignant tumours. Investigations such as fine needle aspiration cytology and MRI scans provide some useful information, but most cases will require surgical excision as a means of coming to a definitive diagnosis. Benign tumours and early low-grade malignancies can be adequately treated with surgery alone, while more advanced and high-grade tumours with regional lymph node metastasis will require postoperative radiotherapy. The role of chemotherapy remains largely palliative. This paper highlights some of the more important aspects in the management of salivary gland tumours. PMID:23724273

  15. [The value of ultrasonography in salivary pathology].

    PubMed

    Katz, P

    1990-01-01

    Ultrasound has been increasingly used in recent years and thanks to high performance, easy to use apparatus, it can now be used for exploration of the salivary glands. This non invasive, painless and relatively inexpensive examination provides rapid visualisation of the salivary glands and is a useful adjunct to radio-xero-sialographic examination, particularly in tumour pathology. Following a review of normal appearances, the author briefly describes the various salivary gland lesions. PMID:2130478

  16. Salivary Diagnostics: A Brief Review

    PubMed Central

    Malathi, Narasimhan; Mythili, Sabesan; Vasanthi, Hannah R.

    2014-01-01

    Early detection of disease plays a crucial role for treatment planning and prognosis. Saliva has great potential as a diagnostic fluid and offers advantage over serum and other biological fluids by an economic and noninvasive collection method for monitoring of systemic health and disease progression. The plethora of components in this fluid can act as biomarkers for diagnosis of various systemic and local diseases. In this review paper, we have emphasized the role of salivary biomarkers as diagnostic tools. PMID:24616813

  17. Classification of microbial ?-amylases for food manufacturing using proteinase digestion.

    PubMed

    Akiyama, Takumi; Yamazaki, Takeshi; Tada, Atsuko; Ito, Yusai; Otsuki, Noriko; Akiyama, Hiroshi

    2014-09-01

    Enzymes produced by microorganisms and plants are used as food additives to aid the processing of foods. Identification of the origin of these enzyme products is important for their proper use. Proteinase digestion of ?-amylase products, followed by high performance liquid chromatography (HPLC) analysis, was applied to ?-amylase from the mold Aspergillus species, the bacteria Bacillus species, and the actinomycetes Saccharomonospora species. Eighteen commercial products of ?-amylase were digested with trypsin and endoproteinase Lys-C and HPLC analyzed. For some proteinase/sample combinations, the area of the intact ?-amylase peak decreased and new peaks were detected after digestion. The presence and retention times of the novel peaks were used to group the products. The results from this method, called the proteinase digestion-HPLC method, allowed the classification of the ?-amylase products into 10 groups, whereas the results from sodium dodecyl sulfate polyacrylamide gel electrophoresis allowed their classification into seven groups. PMID:25473515

  18. Classification of microbial ?-amylases for food manufacturing using proteinase digestion

    PubMed Central

    Akiyama, Takumi; Yamazaki, Takeshi; Tada, Atsuko; Ito, Yusai; Otsuki, Noriko; Akiyama, Hiroshi

    2014-01-01

    Enzymes produced by microorganisms and plants are used as food additives to aid the processing of foods. Identification of the origin of these enzyme products is important for their proper use. Proteinase digestion of ?-amylase products, followed by high performance liquid chromatography (HPLC) analysis, was applied to ?-amylase from the mold Aspergillus species, the bacteria Bacillus species, and the actinomycetes Saccharomonospora species. Eighteen commercial products of ?-amylase were digested with trypsin and endoproteinase Lys-C and HPLC analyzed. For some proteinase/sample combinations, the area of the intact ?-amylase peak decreased and new peaks were detected after digestion. The presence and retention times of the novel peaks were used to group the products. The results from this method, called the proteinase digestion–HPLC method, allowed the classification of the ?-amylase products into 10 groups, whereas the results from sodium dodecyl sulfate polyacrylamide gel electrophoresis allowed their classification into seven groups. PMID:25473515

  19. Salivary Gland Development: A Template for Regeneration

    PubMed Central

    Patel, Vaishali N.; Hoffman, Matthew P.

    2014-01-01

    The mammalian salivary gland develops as a highly branched structure designed to produce and secrete saliva. This review will focus on research on mouse submandibular gland development and the translation of this basic research towards therapy for patients suffering from salivary hypofunction. Here we review the most recent literature that has enabled a better understanding of the mechanisms of salivary gland development. Additionally, we discuss approaches proposed to restore salivary function using gene and cell-based therapy. Increasing our understanding of the developmental mechanisms involved during development is critical to design effective therapies for regeneration and repair of damaged glands. PMID:24333774

  20. Measurement of salivary cortisol in 2012 - laboratory techniques and clinical indications.

    PubMed

    Inder, Warrick J; Dimeski, Goce; Russell, Anthony

    2012-11-01

    The utility of measuring salivary cortisol has become increasingly appreciated since the early 1980s. Salivary cortisol is a measure of active free cortisol and follows the diurnal rhythm of serum or plasma cortisol. The saliva sample may be collected by drooling or through the use of absorbent swabs which are placed into the mouth until saturated. Salivary cortisol is therefore convenient for patients and research participants to collect noninvasively on an outpatient basis. Several assay techniques have been used to measure salivary cortisol, including radioimmunoassay and more recently liquid chromatography-tandem mass spectrometry. The analytical sensitivity varies between these assay methods, as does the potential for cross-reactivity with other steroids. The interpretation of salivary cortisol levels relies on rigorous standardization of sampling equipment, sampling protocols and assay technology with establishment of a local reference range. Clinically, the commonest use for salivary cortisol is measuring late-night salivary cortisol as a screening test for Cushing's syndrome. Several studies have shown diagnostic sensitivities and specificities of over 90%, which compares very favourably with other screening tests for Cushing's syndrome such as the 24-h urinary-free cortisol and the 1-mg overnight dexamethasone suppression test. There are emerging roles for the use of salivary cortisol in diagnosing adrenal insufficiency, particularly in conditions associated with low cortisol-binding globulin levels, and in the monitoring of glucocorticoid replacement. Finally, salivary cortisol has been used extensively as a biomarker of stress in a research setting, especially in studies examining psychological stress with repeated measurements. PMID:22812714

  1. Purification and characterization of an extracellular alpha-amylase from Clostridium perfringens type A.

    PubMed Central

    Shih, N J; Labbé, R G

    1995-01-01

    An alpha-amylase (EC 3.2.1.1) secreted by Clostridium perfringens NCTC 8679 type A was purified to homogeneity and characterized. It was isolated from concentrated cell-free culture medium by ion-exchange and gel permeation chromatography. The enzyme exhibited maximal activity at pH 6.5 and 30 degrees C without the presence of calcium. The pI of the enzyme was 4.75. The estimated molecular weight of the purified enzyme was 76 kDa. The purified enzyme was inactivated between 35 and 40 degrees C, which increased to between 45 and 50 degrees C in the presence of calcium (5 mM). The purified enzyme produced a mixture of oligosaccharides as major end products of starch hydrolysis, indicating alpha-amylase activity. PMID:7646015

  2. Purification and characterisation of ?-amylase produced by mutant strain of Aspergillus oryzae EMS-18.

    PubMed

    Abdullah, Roheena; Ikram-Ul-Haq

    2015-04-01

    ?-Amylase produced by a mutant strain of Aspergillus oryzae EMS-18 has been purified to homogeneity as judged by sodium dodecyle sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was purified by using 70% ammonium sulphate precipitation followed by anion exchange chromatography on DEAE-Sephadex column and gel filtration on Sephadex G-100. An enzyme purification factor of 9.5-fold was achieved with a final specific activity of 1987.7 U/mg protein and overall yield of 23.8%. The molecular weight of purified ?-amylase was estimated to be 48 kDa by SDS-PAGE. The purified enzyme revealed an optimum assay temperature and pH 40°C and 5.0, respectively. Except Ca(++) all other metal ions such as Mg, Mn, Na, Zn, Ni, Fe, Cu, Co and Ba were found to be inhibitory to enzyme activity. PMID:25424747

  3. Children's Context Inappropriate Anger and Salivary Cortisol

    PubMed Central

    Locke, Robin L.; Davidson, Richard J.; Kalin, Ned H.; Goldsmith, H. Hill

    2009-01-01

    Some children show emotion that is not consistent with normative appraisal of the context and can therefore be defined as context inappropriate (CI). The authors used individual growth curve modeling and hierarchical multiple regression analyses to examine whether CI anger predicts differences in hypothalamic-pituitary-adrenal axis activity, as manifest in salivary cortisol measures. About 23% of the 360 children (ages 6–10 years, primarily 7–8) showed at least 1 expression of CI anger in situations designed to elicit positive affect. Expression of anger across 2 positive assessments was less common (around 4%). CI anger predicted the hypothesized lower levels of cortisol beyond that attributed to context appropriate anger. Boys' CI anger predicted lower morning cortisol and flatter slopes. Results suggest that this novel approach to studying children's emotion across varying contexts can provide insight into affective style. PMID:19702392

  4. Effect of pilocarpine mouthwash on salivary flow

    Microsoft Academic Search

    R. Bernardi; C. Perin; F. L. Becker; G. Z. Ramos; G. Z. Gheno; L. R. Lopes; M. Pires; H. M. T. Barros

    2002-01-01

    Pilocarpine is a cholinergic agonist that increases salivary flow and has been used to treat xerostomia. Oral intake is the most frequent route of administration. Adverse effects are dose-dependent and in- clude sudoresis, facial blushing and increased urinary frequency. The objective of the present study was to evaluate the effects of topical pilocarpine solutions as mouthwashes on salivary flow and

  5. Salivary gland derived peptides as a new class of anti-inflammatory agents: review of preclinical pharmacology of C-terminal peptides of SMR1 protein

    Microsoft Academic Search

    Ronald D Mathison; Joseph S Davison; A Dean Befus; Daniel A Gingerich

    2010-01-01

    The limitations of steroidal and non steroidal anti-inflammatory drugs have prompted investigation into other biologically based therapeutics, and identification of immune selective anti-inflammatory agents of salivary origin. The traditional view of salivary glands as accessory digestive structures is changing as their importance as sources of systemically active immunoregulatory and anti-inflammatory factors is recognized. Salivary gland involvement in maintenance of whole

  6. A model study on Eudragit and polyethyleneimine as soluble carriers of ?-amylase for repeated hydrolysis of starch

    Microsoft Academic Search

    Lina Cong; Rajni Kaul; Ulla Dissing; Bo Mattiasson

    1995-01-01

    Eudragit and polyethyleneimine have been evaluated for their potential as soluble carriers of enzymes acting on macromolecular substrates. The present study is based on coupling of ?-amylase (Termamyl) to these polymers for the degradation of starch. The enzyme was bound to both the polymers by carbodiimide coupling. Using soluble starch as substrate, about 96% of the enzyme activity was found

  7. Biochemical and Structural Characterization of Amy1: An Alpha-Amylase from Cryptococcus flavus Expressed in Saccharomyces cerevisiae

    PubMed Central

    Galdino, Alexsandro Sobreira; Silva, Roberto Nascimento; Lottermann, Muriele Taborda; Álvares, Alice Cunha Morales; de Moraes, Lídia Maria Pepe; Torres, Fernando Araripe Gonçalves; de Freitas, Sonia Maria; Ulhoa, Cirano José

    2011-01-01

    An extracellular alpha-amylase (Amy1) whose gene from Cryptococcus flavus was previously expressed in Saccharomyces cerevisiae was purified to homogeneity (67?kDa) by ion-exchange and molecular exclusion chromatography. The enzyme was activated by NH4+ and inhibited by Cu+2 and Hg+2. Significant biochemical and structural discrepancies between wild-type and recombinant ?-amylase with respect to Km values, enzyme specificity, and secondary structure content were found. Far-UV CD spectra analysis at pH 7.0 revealed the high thermal stability of both proteins and the difference in folding pattern of Amy1 compared with wild-type amylase from C. flavus, which reflected in decrease (10-fold) of enzymatic activity of recombinant protein. Despite the differences, the highest activity of Amy1 towards soluble starch, amylopectin, and amylase, in contrast with the lowest activity of Amy1w, points to this protein as being of paramount biotechnological importance with many applications ranging from food industry to the production of biofuels. PMID:21490699

  8. Tobacco plants transformed with the bean. alpha. ai gene express an inhibitor of insect. alpha. -amylase in their seeds. [Nicotiana tabacum; Tenebrio molitor

    SciTech Connect

    Altabella, T.; Chrispeels, M.J. (Univ. of California, San Diego, La Jolla (USA))

    1990-06-01

    Bean (Phaseolus vulgaris L.) seeds contain a putative plant defense protein that inhibits insect and mammalian but not plant {alpha}-amylases. We recently presented strong circumstantial evidence that this {alpha}-amylase inhibitor ({alpha}Al) is encoded by an already-identified lectin gene whose product is referred to as lectin-like-protein (LLP). We have now made a chimeric gene consisting of the coding sequence of the lectin gene that encodes LLP and the 5{prime} and 3{prime} flanking sequences of the lectin gene that encodes phytohemagglutinin-L. When this chimeric gene was expressed in transgenic tobacco (Nicotiana tabacum), we observed in the seeds a series of polypeptides (M{sub r} 10,000-18,000) that cross-react with antibodies to the bean {alpha}-amylase inhibitor. Most of these polypeptides bind to a pig pancreas {alpha}-amylase affinity column. An extract of the seeds of the transformed tobacco plants inhibits pig pancreas {alpha}-amylase activity as well as the {alpha}-amylase present in the midgut of Tenebrio molitor. We suggest that introduction of this lectin gene (to be called {alpha}ai) into other leguminous plants may be a strategy to protect the seeds from the seed-eating larvae of Coleoptera.

  9. Characterization of a stable intermediate trapped during reversible refolding of Bacillus subtilis alpha-amylase.

    PubMed

    Haddaoui, E A; Leloup, L; Petit-Glatron, M F; Chambert, R

    1997-10-15

    Bacillus subtilis exocellular alpha-amylase is reversibly refolded after denaturation by guanidine hydrochloride at pH 7 and 37 degrees C. The unfolding-folding transition monitored by intrinsic fluorescence changes and resistance to proteolysis was resolved into a two-state transition. The first step (t1/2 < 1 s) led from D, the totally unfolded state, to C, a stable partially structured state of the protein. This folding intermediate was devoid of any enzyme activity and partially resistant to protease degradation. Calcium was required for the transition from C to N, the native state. This metal did not remain associated with the native form and could be replaced by barium or strontium, but not by magnesium. We discuss the hypothesis that C, the folding intermediate whose further transformation is under kinetic control, is the competent state involved in the secretion process of alpha-amylase. PMID:9370360

  10. Purification and characterisation of a novel amylase enzyme from red pitaya (Hylocereus polyrhizus) peel.

    PubMed

    Amid, Mehrnoush; Abd Manap, Mohd Yazid

    2014-12-15

    An amylase enzyme from pitaya peel was purified 234.2-folds with 72.1% recovery using ammonium sulphate precipitation, gel filtration and ion exchange chromatography. Gel filtration chromatography and SDS-PAGE revealed that the enzyme is monomeric with a molecular weight of 42.1kDa. The apparent Km and Vmax of the amylase were 2.7 mg/ml and 34.30 u/min/mg of protein, respectively. The enzyme was highly active and stable over a wide pH range from pH 3 to pH 11.0, with optimum activity being observed at pH 5.0. The enzyme was highly selective for soluble starch, amylopectin, glycogen and pulullan. The purified amylase did not require calcium and displayed extreme stability with regard to surfactants and oxidising agents. EDTA, a powerful chelating agent, did not have any significant effect on the stability of the enzyme. Such characteristics have not been previously reported for this type of enzyme from fruit peel. This enzyme, which possesses unique properties, could be widely used in different types of industries, especially in food and biotechnological applications. PMID:25038694

  11. Cloning and Characterization of an alpha-amylase Gene from the Hyperthermophilic Archaeon Thermococcus Thioreducens

    NASA Technical Reports Server (NTRS)

    Bernhardsdotter, Eva C. M. J.; Pusey, Mark L.; Ng, Joseph D.; Garriott, Owen K.

    2004-01-01

    The gene encoding an extracellular alpha-amylase, TTA, from the hyperthermophilic archaeon Thermococcus thioreducens was cloned and expressed in Escherichia coli. Primary structural analysis revealed high similarity with other a-amylases from the Thermococcus and Pyrococcus genera, as well as the four highly conserved regions typical for a-amylases. The 1374 bp gene encodes a protein of 457 amino acids, of which 435 constitute the mature protein preceded by a 22 amino acid signal peptide. The molecular weight of the purified recombinant enzyme was estimated to be 43 kDa by denaturing gel electrophoresis. Maximal enzymatic activity of recombinant TTA was observed at 90 C and pH 5.5 in the absence of exogenous Ca(2+), and the enzyme was considerably stable even after incubation at 90 C for 2 hours. The thermostability at 90 and 102 C was enhanced in the presence of 5 mM Ca(2+). The extraordinarily high specific activity (about 7.4 x 10(exp 3) U/mg protein at 90 C, pH 5.5 with soluble starch as substrate) together with its low pH optimum makes this enzyme an interesting candidate for starch processing applications.

  12. Cloning and Characterization of an Alpha-amylase Gene from the Hyperthermophilic Archaeon Thermococcus Thioreducens

    NASA Technical Reports Server (NTRS)

    Bernhardsdotter, Eva C. M. J.; Pusey, Marc L.; Ng, Joseph D.; Garriott, Owen K.

    2004-01-01

    The gene encoding an extracellular a-amylase, TTA, from the hyperthermophilic archaeon Thermococcus thioreducens was cloned and expressed in Escherichia coli. Primary structural analysis revealed high similarity with other a-amylases from the Thermococcus and Pyrococcus genera, as well as the four highly conserved regions typical for a-amylases. The 1374 bp gene encodes a protein of 457 amino acids, of which 435 constitute the mature protein preceded by a 22 amino acid signal peptide. The molecular weight of the purified recombinant enzyme was estimated to be 43 kDa by denaturing gel electrophoresis. Maximal enzymatic activity of recombinant TTA was observed at 90 C and pH 5.5 in the absence of exogenous Ca(2+), and the enzyme was considerably stable even after incubation at 90 C for 2 hours. The thermostability at 90 and 102 C was enhanced in the presence of 5 mM Ca(2+). The extraordinarily high specific activity (about 7.4 x 10(exp 3) U/mg protein at 90 C, pH 5.5 with soluble starch as substrate) together with its low pH optimum makes this enzyme an interesting candidate for starch processing applications.

  13. Iron binding modulates candidacidal properties of salivary histatin 5.

    PubMed

    Puri, S; Li, R; Ruszaj, D; Tati, S; Edgerton, M

    2015-01-01

    Salivary protein histatin 5 (Hst 5) is fungicidal toward Candida albicans, the causative agent of oropharyngeal candidiasis. However, its activity in saliva is compromised by salivary protease-mediated degradation and interaction with salivary salts. Hst 5 has also been shown to bind various metals in saliva-namely, Zn, Cu, and Ni. Surprisingly, interactions of Hst 5 with Fe have not been studied, although iron is one of the most abundant metals present in saliva. Using circular dichroism, we show that Hst 5 can bind up to 10 equivalents of iron as measured by loss of its alpha-helical secondary structure that is normally observed for it in trifluoroethylene. A significant decrease in the candidacidal ability of Hst 5 was observed upon iron binding, with increasing iron concentrations being inversely proportional to Hst 5 killing activity. Binding assays showed that the decrease in killing was likely a result of reduced binding (10-fold reduction) of Fe-Hst 5 to C. albicans cells. Protease stability analysis showed that Fe-Hst 5 was completely resistant to trypsin digestion. In contrast, zinc binding had limited effects on Hst 5 fungicidal activity or protease susceptibility. RNA sequencing results identified changes in iron uptake genes in Hst 5-treated C. albicans cells. Our findings thus suggest that consequences of Hst 5 binding iron not only affect candidacidal ability and proteolyic stability of Hst 5, but may also contribute to a novel killing mechanism involving interference with cellular iron metabolism. PMID:25365968

  14. Characterisation of three starch degrading enzymes: thermostable ?-amylase, maltotetraogenic and maltogenic ?-amylases.

    PubMed

    Derde, L J; Gomand, S V; Courtin, C M; Delcour, J A

    2012-11-15

    Maltogenic ?-amylase from Bacillus stearothermophilus (BStA) is widely used as bread crumb anti-firming enzyme. A maltotetraose-forming ?-amylase from Pseudomonas saccharophila (PSA) was recently proposed as alternative, hence the need to compare both exo-acting enzymes with some endo-action component. A purely exo-acting thermostable ?-amylase from Clostridium thermosulfurogenes (CTB) was included for reference purposes. Under the experimental conditions used, temperature optima of the enzymes are rather similar (60-65 °C), but temperature stability decreased in the order BStA, PSA and CTB. The action of the enzymes on different substrates and their impact on the rheological behaviour of maize starch suspensions demonstrated that, while CTB acts exclusively through an exo-action mechanism, BStA displayed limited endo-action which became more pronounced at higher temperatures. PSA has more substantial endo-action than BStA, which is rather temperature independent. This is important for their impact in processes such as breadmaking, where temperature is gradually increased. PMID:22868150

  15. Ornithodoros brasiliensis (mouro tick) salivary gland homogenates inhibit in vivo wound healing and in vitro endothelial cell proliferation.

    PubMed

    Reck, José; Marks, Fernanda S; Termignoni, Carlos; Guimarães, Jorge A; Martins, João Ricardo

    2013-04-01

    Ornithodoros brasiliensis is a nidicolous tick only found in the southern Brazilian highlands region. O. brasiliensis parasitism is frequently associated with toxicosis syndrome, which can lead to severe reactions, ranging from local pruritus and pain to systemic disturbances both in humans and dogs. One of the most frequent findings associated with an O. brasiliensis bite is a slow healing lesion at the site of tick attachment, which can take several weeks to heal. This work tested the hypothesis that an O. brasiliensis salivary gland homogenate is able to modulate the skin wound-healing process in vivo, using a model of excisional skin lesion in rats, which are divided into two groups: (1) control group and (2) treated group, which topically received salivary gland homogenate equivalent to the protein amount of one whole salivary gland (?5 ?g protein). The hypothesis that O. brasiliensis salivary gland homogenates interfere with endothelial cell proliferation, a key role phenomenon in wound healing, was also tested. O. brasiliensis salivary gland homogenates significantly delay skin wound healing. The time to full healing of skin lesions in control rats was 15 days, contrasting with 24 days in rats topically treated with O. brasiliensis salivary gland homogenates. The calculated HT50 (healing time to recover 50% of the wound area) for control groups was 3.6 days (95% CI, 3.2-3.9) and for salivary gland treated rats was 7.7 days (95% CI, 7.0-8.4). Salivary gland homogenates have a strong cytotoxic activity on cultured endothelial cells (LC50, 13.6 mg/ml). Also, at sublethal concentrations (?3 mg/ml), salivary gland homogenates have a remarkable anti-proliferative activity (IC50 0.7 mg/ml) on endothelial cells, equivalent to ?0.03 salivary gland pairs, an activity which seems to be much greater than reported for any other tick species. This is the first report about the biological activities of O. brasiliensis salivary compounds and provides the first in vivo evidence to support the concept of wound-healing modulation by tick salivary secretions. Results shown here contribute to an understanding of O. brasiliensis tick toxicosis syndrome, and also increase our knowledge of tick salivary bioactive compounds. PMID:23397378

  16. The Association between Midnight Salivary Cortisol and Metabolic Syndrome in Korean Adults

    PubMed Central

    Jang, Yun-Mi; Lee, Eun Jung; Kim, Dong Lim; Kim, Suk Kyeong

    2012-01-01

    Background The common characteristics of metabolic syndrome (MetS) and Cushing's syndrome suggest that excess cortisol may be involved in the pathogenesis of MetS. Salivary cortisol measurements are simple and can be surrogates for plasma free cortisol, which is the most biologically active form. We evaluated the association between levels of midnight salivary cortisol and MetS in Korean adults. Methods A total of 46 subjects, aged 20 to 70 years, who visited the Health Care Center at Konkuk University Hospital from August 2008 to August 2009 were enrolled. We compared the levels of midnight salivary cortisol in subjects with MetS with those in subjects without MetS. We analyzed the associations between midnight salivary cortisol levels and components of MetS. Results Midnight salivary cortisol levels were higher in the MetS group (70±42.4 ng/dL, n=12) than that in the group without MetS (48.1±36.8 ng/dL, n=34) (P=0.001). Positive correlations were observed between midnight salivary cortisol levels and waist circumference, fasting blood glucose, and homeostasis model assessment of insulin resistance. The risk for MetS was significantly higher in subjects with midnight salivary cortisol levels ?100 ng/dL than in those with levels <50 ng/dL (odds ratio, 5.9; 95% confidence interval, 2.35 to 36.4). Conclusion The results showed a positive correlation between midnight salivary cortisol levels and MetS, suggesting that hypercortisolism may be related to MetS. PMID:22737665

  17. Rapid preparative isolation of erythrocentaurin from Enicostemma littorale by medium-pressure liquid chromatography, its estimation by high-pressure thin-layer chromatography, and its ?-amylase inhibitory activity.

    PubMed

    Hassan, Naila; Ahamad, Javed; Amin, Saima; Mujeeb, Mohd; Mir, Showkat R

    2015-02-01

    Erythrocentaurin is a relatively simple natural product present among the members of Gentianaceae. A preparative method for the isolation of erythrocentaurin from the ethyl acetate fraction of Enicostemma littorale using medium-pressure liquid chromatography has been reported. The method consisted of a simple step gradient from 10 to 20% ethyl acetate in n-hexane. Using a 70 × 460 mm Si60 column, this method is capable of processing 20 g of material in <3 h (purity ? 97%). The recovery of erythrocentaurin was 87.77%. Estimation of erythrocentaurin in extracts and fractions based on high-pressure thin-layer chromatography was carried out on silica gel 60 F(254) plates with toluene/ethyl acetate/formic acid (80:18:2 v/v/v) as the mobile phase. The densitometric analysis was performed at 230 nm. A well-separated compact band of erythrocentaurin appeared at R(f )0.54 ± 0.04. The analytical method showed good linearity in the concentration range of 200-1500 ng/band with a correlation coefficient of 0.99417. The limits of detection and quantification were found to be ?60 and ?180 ng/band, respectively. Erythrocentaurin exhibited a concentration-dependent ?-amylase inhibition (IC(50) 1.67 ± 0.28 mg/mL). The outcome of the study should be considered for pharmacokinetic and biotransformation studies involving E. littorale. PMID:25504557

  18. SOURCES OF LARVAL SALIVARY GLAND SECRETION IN THE DIPTERAN CHIRONOMUS TENTANS

    PubMed Central

    Doyle, D.; Laufer, H.

    1969-01-01

    The soluble proteins in the hemolymph, the salivary gland, and the salivary secretion of fourth instar Chironomus tentans were examined by disc electrophoresis in acrylamide gels. Of the 11 protein fractions detected in buffered saline extracts of the gland, 10 are present also in the hemolymph. Amino acid isotope incorporation experiments indicate that the protein fractions shared by the salivary gland and the hemolymph are not synthesized in the gland but are synthesized in other larval tissues. Immunochemical studies show that most of these proteins eventually are secreted from the gland. The salivary gland in vivo and in vitro is active in de novo protein synthesis. The protein synthesized tends to form large molecular weight aggregates. As demonstrated by radioautography, at least 80% of this protein is secreted from the 30 large cells forming most of the gland. The proteins synthesized in the salivary gland cannot be detected in the hemolymph. The results of this investigation are consistent with a mechanism of secretion formation involving both de novo synthesis of some secretion proteins and the selective uptake, transport, and secretion of hemal proteins by the salivary gland. PMID:5782452

  19. A Role for Notch Signaling in Salivary Acinar Cell Growth and Differentiation

    PubMed Central

    Dang, Howard; Lin, Alan L; Zhang, Binxian; Zhang, Hong-Mei; Katz, Michael S; Yeh, Chih-Ko

    2009-01-01

    The Notch pathway is crucial for stem/progenitor cell maintenance, growth and differentiation in a variety of tissues. The Notch signaling is essential for Drosophila salivary gland development but its role in mammalian salivary gland remains unclear. The human salivary epithelial cell line, HSG, was studied to determine the role of Notch signaling in salivary epithelial cell differentiation. HSG expressed Notch 1 to 4, and the Notch ligands Jagged 1 and 2 and Delta 1. Treatment of HSG cells with inhibitors of ?-secretase, which is required for Notch cleavage and activation, blocked vimentin expression, an indicator of HSG differentiation. HSG differentiation was also associated with Notch downsteam signal Hes-1 expression, and Hes-1 expression was inhibited by ?-secretase inhibitors. siRNA corresponding to Notch 1 to 4 was used to show that silencing of all four Notch receptors was required to inhibit HSG differentiation. Normal human submandibular gland expressed Notch 1 to 4, Jagged 1 and 2, and Delta 1, with nuclear localization indicating Notch signaling in vivo. Hes-1 was also expressed in the human tissue, with staining predominantly in the ductal cells. In salivary tissue from rats undergoing and recovering from ductal obstruction, we found that Notch receptors and ligands were expressed in the nucleus of the regenerating epithelial cells. Taken together, these data suggest that Notch signaling is critical for normal salivary gland cell growth and differentiation. PMID:19235730

  20. Salivary defense factors in herpes simplex virus infection.

    PubMed

    Välimaa, H; Waris, M; Hukkanen, V; Blankenvoorde, M F J; Nieuw Amerongen, A V; Tenovuo, J

    2002-06-01

    Saliva may contribute to a lowering of the infectious herpes simplex virus (HSV) dose during transmission and consequently abrogate infection or lead to decreased reactivation. To test this hypothesis, we assayed saliva for innate defense factors, immunoglobulin content, and the capacity to interfere with HSV infection. Serum or salivary anti-HSV IgG levels did not correlate with control of recurrent labial herpes (RLH) and were significantly higher in subjects with RLH compared with asymptomatic seropositive subjects. Although no differences in levels or output rate of innate defense factors between the groups were observed, the salivary neutralizing activity correlated with lactoferrin and hypothiocyanite concentrations in the asymptomatic seropositive group. Our results suggest that saliva contains factors, in addition to anti-HSV immunoglobulins, that neutralize HSV and may indirectly contribute to the control of RLH. PMID:12097435

  1. 21 CFR 862.1070 - Amylase test system.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Chemistry Test Systems § 862.1070 Amylase test system. (a) Identification. An...

  2. Iatrogenic causes of salivary gland dysfunction

    SciTech Connect

    Schubert, M.M.; Izutsu, K.T.

    1987-02-01

    Saliva is important for maintaining oral health and function. There are instances when medical therapy is intended to decrease salivary flow, such as during general anesthesia, but most instances of iatrogenic salivary gland dysfunction represent untoward or unavoidable side-effects. The clinical expression of the salivary dysfunction can range from very minor transient alteration in saliva flow to a total loss of salivary function. The most common forms of therapy that interfere with salivation are drug therapies, cancer therapies (radiation or chemotherapy), and surgical therapy. These therapies can affect salivation by a number of different mechanisms that include: disruption of autonomic nerve function related to salivation, interference with acinar or ductal cell functions related to salivation, cytotoxicity, indirect effects (vasoconstriction/dilation, fluid and electrolyte balance, etc.), and physical trauma to salivary glands and nerves. A wide variety of drugs is capable of increasing or decreasing salivary flow by mimicking autonomic nervous system actions or by directly acting on cellular processes necessary for salivation: drugs can also indirectly affect salivation by altering fluid and electrolyte balance or by affecting blood flow to the glands. Ionizing radiation can cause permanent damage to salivary glands, damage that is manifest as acinar cell destruction with subsequent atrophy and fibrosis of the glands. Cancer chemotherapy can cause changes in salivation, but the changes are usually much less severe and only transient. Finally, surgical and traumatic injuries interfere with salivation because of either disruption of gland innervation or gross physical damage (or removal) of glandular tissue (including ducts).

  3. Cloning, expression, and purification of insect (Sitophilus oryzae) alpha-amylase, able to digest granular starch, in Yarrowia lipolytica host.

    PubMed

    Celi?ska, Ewelina; Bia?as, Wojciech; Borkowska, Monika; Grajek, W?odzimierz

    2015-03-01

    Raw-starch-digesting enzymes (RSDE) are of major importance for industrial applications, as their usage greatly simplifies the starch processing pipeline. To date, only microbial RSDE have gained considerable attention, since only microbial production of enzymes meets industrial demands. In this study, ?-amylase from rice weevil (Sitophilus oryzae), the major rice pest, was cloned and expressed in Yarrowia lipolytica Po1g strain. The enzyme was secreted into the culture medium, and the peak activity (81 AU/L) was reached after only 29 h of culturing in 5-L bioreactors. Through simple purification procedure of ammonium sulfate precipitation and affinity chromatography, it was possible to purify the enzyme to apparent homogeneity (25-fold purification factor, at 5 % yield). The optimal conditions for the ?-amylase activity were pH 5.0 and a temperature of 40 °C. The ?-amylase studied here did not show any obligate requirement for Ca(2+) ions. The recombinant ?-amylase appeared to efficiently digest granular starch from pea, amaranth, waxy corn, and waxy rice. PMID:25547839

  4. Studies of the effect of maltose on the direct binding of porcine pancreatic ?-amylase to maize starch.

    PubMed

    Warren, Frederick J; Butterworth, Peter J; Ellis, Peter R

    2012-09-01

    For a two phase system comprising an enzyme in solution acting on an insoluble substrate such as starch, adsorption of the enzyme is a key initial step in the reaction but few studies of agents affecting direct binding have been performed. The effect of maltose on the interaction of maize starch with porcine pancreatic ?-amylase was studied by using a method in which the direct binding of starch to amylase is measured under conditions of negligible catalytic activity. The dissociation constant for starch binding increased with maltose concentration and analysis of the binding showed that the kinetic action of maltose was entirely competitive. This result accords with results described in the literature in which maltose was shown to be a competitive inhibitor of amylase action. If the maltose concentration is sufficiently high, a second molecule may bind at the active site but the affinity of the second binding step is approximately 6.5-fold weaker. Because of the relatively low affinity for maltose, it seems unlikely that inhibition by maltose of the initial stage of starch-amylase interaction normally plays any role in regulating intestinal digestion of starch. PMID:22867906

  5. Molecular cloning and expression of an alpha-amylase inhibitor from rye with potential for controlling insect pests.

    PubMed

    Dias, Simoni C; Franco, Octávio L; Magalhães, Cláudio P; de Oliveira-Neto, Osmundo B; Laumann, Raúl A; Figueira, Edson L Z; Melo, Francislete R; Grossi-De-Sá, Maria F

    2005-02-01

    Alpha-amylase inhibitors have important roles in plant defense mechanisms, particularly against insects, and several of these inhibitors have been expressed in different crops to increase their resistance to particular insects. In this work, we report the cloning and expression of a gene encoding for a new alpha-amylase inhibitor (BIII) from rye (Secale cereale) seeds. The BIII gene contains 354 nucleotides that encode for 118 amino acids sequence. A 313 bp fragment of the gene was expressed in Escherichia coli and resulted in a functional inhibitor that reduced the activity of alpha-amylases of larvae of the coleopteran pests Acanthoscelides obtectus, Zabrotess subfasciatus and Anthonomus grandis. In contrast, the inhibitor did not inhibit the activity of porcine pancreatic alpha-amylase. Although the amino acid sequence of BIII showed high identity with those of bifunctional inhibitors, the recombinant protein was unable to inhibit trypsin-like serine proteinases. The effects of recombinant BIII were evaluated in vivo against A. grandis. When first instar larvae were reared on an artificial diet containing four different concentrations of BIII, a reduction in larval weight and a mortality of 83% were observed at the highest concentration. PMID:16003953

  6. Engineering of the alpha-amylase from Geobacillus stearothermophilus US100 for detergent incorporation.

    PubMed

    Khemakhem, Bassem; Ali, Mamdouh Ben; Aghajari, Nushin; Juy, Michel; Haser, Richard; Bejar, Samir

    2009-02-01

    AmyUS100DeltaIG is a variant of the most thermoactive and thermostable maltohexaose forming alpha-amylase produced by Geobacillus stearothermophilus sp.US100. This enzyme which was designed to improve the thermostability of the wild-type enzyme has acquired a very high resistance to chelator agents. According to modeling structural studies and with the aim of enhancing its resistance towards chemical oxidation, a mutant (AmyUS100DeltaIG/M197A) was created by substituting methionine 197 to alanine. The catalytic proprieties of the resulting mutant show alterations in the specific activity and the profile of starch hydrolysis. Interestingly, AmyUS100DeltaIG/M197A displayed the highest resistance to oxidation compared to the AmyUS100DeltaIG and to Termamyl300, the well-known commercial amylase used in detergent. Further, performance of the engineered alpha-amylase was estimated in the presence of commonly used detergent compounds and a wide range of commercial detergent (liquid and solid). These studies indicated a high compatibility and performance of AmyUS100DeltaIG/M197A, suggesting its potential application in detergent industry. PMID:18951544

  7. [An amylase from fresh fruiting bodies of the monkey head mushroom Hericium erinaceum].

    PubMed

    Du, F; Wang, H X; Ng, T B

    2013-01-01

    An amylase with a molecular mass of 55 kDa and an N-terminal sequence exhibiting similarity to enzyme from Bacteroides thetaitaomicron was isolated from fruiting bodies of the monkey head mushroom Hericium erinaceum. The purification scheme included extraction with distilled water, ion exchange chromatography on DEAE-cellulose and SP-sepharose, and gel filtration by FPLC on Superdex 75. The amylase of H. erinaceum was adsorbed on DEAE-cellulose in 10 mM Tris-HCl buffer (pH 7.4) and eluted with 0.2 M NaCl in the same buffer. The enzyme was subsequently adsorbed on SP-Sepharose in 10 mM ammonium acetate buffer (pH 4.5) and eluted with 0.3 M NaCl in the same buffer. This fraction was subsequently subjected to gel filtration on Superdex 75. The first peak eluted had a molecular mass of 55 kDa in SDS-PAGE. The amylase of H. erinaceum exhibited a pH optimum of 4.6 and a temperature optimum of 40 degrees C. The enzyme activity was enhanced by Mn2+ and Fe3+ ions, but inhibited by Hg2+ ions. PMID:23662447

  8. Alpha amylase enzyme inhibitory and anti-inflammatory effect of Lawsonia inermis.

    PubMed

    Imam, Hasan; Mahbub, Nasir Uddin; Khan, Md Forhad; Hana, Humayera Kabir; Sarker, Md Moklesur Rahman

    2013-12-01

    Previously it was reported elsewhere that Lawsonia inermis have anti-inflammatory and analgesic effect in experimental animals. The in vitro porcine alpha amylase inhibitory effect was investigated of this plant methanolic extracts and consequently hypoglycemic effect by quantitatively determining the maltose from the maltose standard curve while the anti-inflammatory effect by acetic acid induced writhing test in mice. Acarbose (10 microg mL(-1)) and Diclofenac sodium (20 mg kg(-1)) were used as reference hypoglycemic and anti-inflammatory drugs, respectively, for this study. The methanolic leaves extract of the plant significantly inhibited (60.97% compared to untreated) enzymatic activity of the amylase at 10 microg mL(-1) dose (p < 0.05) also reduced the chemically induced nociceptive pain stimuli significantly at all doses (p < 0.01). Carbohydrates, glycosides, flavonoids, saponins and tannins were found to have in phytochemical screening of the extract which are thought to bring these effects. For the conclusive purpose, it is suggesting from the result that the pharmacological properties of this Lawsonia inermis can elicit hypoglycemic effect by inhibiting alpha-amylase enzyme and can reduce neurogenic pain stimulus. It gives the notion that how this group of patient would be therapeutically benefitted by decreasing both these effects by the same agent which is easy available. PMID:24506051

  9. Application of decolourized and partially purified polygalacturonase and ?-amylase in apple juice clarification

    PubMed Central

    Dey, Tapati Bhanja; Banerjee, Rintu

    2014-01-01

    Polygalacturonase and ?-amylase play vital role in fruit juice industry. In the present study, polygalacturonase was produced by Aspergillus awamori Nakazawa MTCC 6652 utilizing apple pomace and mosambi orange (Citrus sinensis var mosambi) peels as solid substrate whereas, ?-amylase was produced from A. oryzae (IFO-30103) using wheat bran by solid state fermentation (SSF) process. These carbohydrases were decolourized and purified 8.6-fold, 34.8-fold and 3.5-fold, respectively by activated charcoal powder in a single step with 65.1%, 69.8% and 60% recoveries, respectively. Apple juice was clarified by these decolourized and partially purified enzymes. In presence of 1% polygalacturonase from mosambi peels (9.87 U/mL) and 0.4% ?-amylase (899 U/mL), maximum clarity (%T660nm = 97.0%) of juice was attained after 2 h of incubation at 50 °C in presence of 10 mM CaCl2. Total phenolic content of juice was reduced by 19.8% after clarification, yet with slightly higher %DPPH radical scavenging property. PMID:24948919

  10. Gingival salivary gland choristoma. A case report.

    PubMed

    Ledesma-Montes, C; Fernández-López, R; Garcés-Ortíz, M; Portilla-Robertson, J; Hernández-Guerrero, J C

    1998-10-01

    Gingival salivary gland choristoma is an extremely rare disturbance of glandular development. A review of the literature disclosed only 5 reported cases of this entity and 7 gingival salivary gland tumors or alterations. We present a case of this condition present in a 43-year-old female patient, which was found while reviewing casts for the design of a prosthetic appliance. This case suggests that embryonal pluripotentiality of gingival epithelial cells is retained and that development of salivary glands in gingival tissue is feasible. An additional discussion about its histogenesis is presented. PMID:9802717

  11. Receptors for the neuropeptides, myoinhibitory peptide and SIFamide, in control of the salivary glands of the blacklegged tick Ixodes scapularis.

    PubMed

    Simo, Ladislav; Ko?i, Juraj; Park, Yoonseong

    2013-04-01

    Tick salivary glands are important organs that enable the hematophagous feeding of the tick. We previously described the innervation of the salivary gland acini types II and III by a pair of protocerebral salivary gland neurons that produce both myoinhibitory peptide (MIP) and SIFamide (Šimo et al., 2009b). In this study we identified authentic receptors expressed in the salivary glands for these neuropeptides. Homology-based searches for these receptors in the Ixodes scapularis genome sequence were followed by gene cloning and functional expression of the receptors. Both receptors were activated by low nanomolar concentrations of their respective ligands. The temporal expression patterns of the two ligands and their respective receptors suggest that the SIFamide signaling system pre-exists in unfed salivary glands, while the MIP system is activated upon initiation of feeding. Immunoreactivity for the SIFamide receptor in the salivary gland was detected in acini types II and III, surrounding the acinar valve and extending to the basal region of the acinar lumen. The location of the SIFamide receptor in the salivary glands suggests three potential target cell types and their probable functions: myoepithelial cell that may function in the contraction of the acini and/or the control of the valve; large, basally located dopaminergic granular cells for regulation of paracrine dopamine; and neck cells that may be involved in the control of the acinar duct and its valve. PMID:23357681

  12. Analysis of salivary gland transcripts of the sand fly Lutzomyia ayacuchensis, a vector of Andean-type cutaneous leishmaniasis.

    PubMed

    Kato, Hirotomo; Jochim, Ryan C; Gomez, Eduardo A; Uezato, Hiroshi; Mimori, Tatsuyuki; Korenaga, Masataka; Sakurai, Tatsuya; Katakura, Ken; Valenzuela, Jesus G; Hashiguchi, Yoshihisa

    2013-01-01

    The saliva of blood sucking insects contains potent pharmacologically active components that assist them in counteracting the host hemostatic and inflammatory systems during blood feeding. In addition, sand fly salivary proteins affect host immunity and have the potential to be a vaccine against Leishmania infection. In the present study, the salivary gland transcripts of Lutzomyia ayacuchensis, a vector of cutaneous leishmaniasis in Ecuadorian and Peruvian Andes, were analyzed by sequencing randomly selected clones of the salivary gland cDNA library of this sand fly. This resulted in the identification of the most abundant transcripts coding for secreted proteins. These proteins were homologous to the salivary molecules present in other sand flies including the RGD-containing peptide, PpSP15/SL1 family protein, yellow-related protein, putative apyrase, antigen 5-related protein, D7 family protein, and 27 kDa salivary protein. Of note, homologues of maxadilan, an active vasodilator abundantly present in saliva of Lutzomyia longipalpis, were not identified. This analysis is the first description of salivary proteins from a sand fly of the subgenus Helcocyrtomyia and from vector of cutaneous leishmaniasis in the New World. The present analysis will provide further insights into the evolution of salivary components in blood sucking arthropods. PMID:23000112

  13. Receptors for the Neuropeptides, Myoinhibitory Peptide and SIFamide, in Control of the Salivary Glands of the Blacklegged Tick Ixodes scapularis

    PubMed Central

    Šimo, Ladislav; Ko?i, Juraj; Park, Yoonseong

    2013-01-01

    Tick salivary glands are important organs that enable the hematophagous feeding of the tick. We previously described the innervation of the salivary gland acini types II and III by a pair of protocerebral salivary gland neurons that produce both myoinhibitory peptide (MIP) and SIFamide (Šimo et al., 2009b). In this study we identified authentic receptors expressed in the salivary glands for these neuropeptides. Homology-based searches for these receptors in the Ixodes scapularis genome sequence were followed by gene cloning and functional expression of the receptors. Both receptors were activated by low nanomolar concentrations of their respective ligands. The temporal expression patterns of the two ligands and their respective receptors suggest that the SIFamide signaling system pre-exists in unfed salivary glands, while the MIP system is activated upon initiation of feeding. Immunoreactivity for the SIFamide receptor in the salivary gland was detected in acini types II and III, surrounding the acinar valve and extending to the basal region of the acinar lumen. The location of the SIFamide receptor in the salivary glands suggests three potential target cell types and their probable functions: myoepithelial cells that may function in the contraction of the acini and/or the control of the valve; large, basally located dopaminergic granular cells for regulation of paracrine dopamine; and neck cells that may be involved in the control of the acinar duct and its valve. PMID:23357681

  14. C-type natriuretic peptide enhances amylase release through NPR-C receptors in the exocrine pancreas.

    PubMed

    Sabbatini, María E; Rodríguez, Myrian; di Carlo, María B; Davio, Carlos A; Vatta, Marcelo S; Bianciotti, Liliana G

    2007-11-01

    Several studies show that C-type natriuretic peptide (CNP) has a modulatory role in the digestive system. CNP administration reduces both jejunal fluid and bile secretion in the rat. In the present study we evaluated the effect of CNP on amylase release in isolated pancreatic acini as well as the receptors and intracellular pathways involved. Results showed that all natriuretic peptide receptors were expressed not only in the whole pancreas but also in isolated pancreatic acini. CNP stimulated amylase secretion with a concentration-dependent biphasic response; maximum release was observed at 1 pM CNP, whereas higher concentrations gradually attenuated it. The response was mimicked by a selective natriuretic peptide receptor (NPR-C) agonist and inhibited by pertussis toxin, strongly supporting NPR-C receptor activation. CNP-evoked amylase release was abolished by U-73122 (PLC inhibitor) and 2-aminoethoxydiphenyl borate (2-APB) [an inositol 1,4,5-triphosphate (IP(3)) receptor antagonist], partially inhibited by GF-109203X (PKC inhibitor), and unaltered by ryanodine or protein kinase A (PKA) and protein kinase G (PKG) inhibitors. Phosphoinositide hydrolysis was enhanced by CNP at all concentrations and abolished by U-73122. At 1 and 10 pM, CNP did not affect cAMP or guanosine 3',5'-cyclic monophosphate (cGMP) levels, but at higher concentrations it increased cGMP and diminished cAMP content. Present findings show that CNP stimulated amylase release through the activation of NPR-C receptors coupled to the PLC pathway and downstream effectors involved in exocytosis. The attenuation of amylase release was likely related to cAMP reduction. The augmentation in cGMP supports activation of NPR-A/NPR-B receptors probably involved in calcium influx. Present findings give evidence that CNP is a potential direct regulator of pancreatic function. PMID:17702953

  15. Comparative analyses of salivary proteins from three aphid species.

    PubMed

    Vandermoten, S; Harmel, N; Mazzucchelli, G; De Pauw, E; Haubruge, E; Francis, F

    2014-02-01

    Saliva is a critical biochemical interface between aphids and their host plants; however, the biochemical nature and physiological functions of aphid saliva proteins are not fully elucidated. In this study we used a multidisciplinary proteomics approach combining liquid chromatography-electrospray ionization tandem mass spectrometry and two-dimensional differential in-gel electrophoresis/matrix-assisted laser desorption/ionization time-of-flight/mass spectrometry to compare the salivary proteins from three aphid species including Acyrthosiphon pisum, Megoura viciae and Myzus persicae. Comparative analyses revealed variability among aphid salivary proteomes. Among the proteins that varied, 22% were related to DNA-binding, 19% were related to GTP-binding, and 19% had oxidoreductase activity. In addition, we identified a peroxiredoxin enzyme and an ATP-binding protein that may be involved in the modulation of plant defences. Knowledge of salivary components and how they vary among aphid species may reveal how aphids target plant processes and how the aphid and host plant interact. PMID:24382153

  16. Protective mechanism of the Mexican bean weevil against high levels of alpha-amylase inhibitor in the common bean.

    PubMed Central

    Ishimoto, M; Chrispeels, M J

    1996-01-01

    Alpha-amylase inhibitor (alpha AI) protects seeds of the common bean (Phaseolus vulgaris) against predation by certain species of bruchids such as the cowpea weevil (Callosobruchus maculatus) and the azuki bean weevil (Callosobruchus chinensis), but not against predation by the bean weevil (Acanthoscelides obtectus) or the Mexican bean weevil (Zabrotes subfasciatus), insects that are common in the Americas. We characterized the interaction of alpha AI-1 present in seeds of the common bean, of a different isoform, alpha AI-2, present in seeds of wild common bean accessions, and of two homologs, alpha AI-Pa present in seeds of the tepary bean (Phaseolus acutifolius) and alpha AI-Pc in seeds of the scarlet runner bean (Phaseolus coccineus), with the midgut extracts of several bruchids. The extract of the Z. subfasciatus larvae rapidly digests and inactivates alpha AI-1 and alpha AI-Pc, but not alpha AI-2 or alpha AI-Pa. The digestion is caused by a serine protease. A single proteolytic cleavage in the beta subunit of alpha AI-1 occurs at the active site of the protein. When degradation is prevented, alpha AI-1 and alpha AI-Pc do not inhibit the alpha-amylase of Z. subfasciatus, although they are effective against the alpha-amylase of C. chinensis. Alpha AI-2 and alpha AI-Pa, on the other hand, do inhibit the alpha-amylase of Z. subfasciatus, suggesting that they are good candidates for genetic engineering to achieve resistance to Z. subfasciatus. PMID:8787024

  17. Protective mechanism of the Mexican bean weevil against high levels of alpha-amylase inhibitor in the common bean.

    PubMed

    Ishimoto, M; Chrispeels, M J

    1996-06-01

    Alpha-amylase inhibitor (alpha AI) protects seeds of the common bean (Phaseolus vulgaris) against predation by certain species of bruchids such as the cowpea weevil (Callosobruchus maculatus) and the azuki bean weevil (Callosobruchus chinensis), but not against predation by the bean weevil (Acanthoscelides obtectus) or the Mexican bean weevil (Zabrotes subfasciatus), insects that are common in the Americas. We characterized the interaction of alpha AI-1 present in seeds of the common bean, of a different isoform, alpha AI-2, present in seeds of wild common bean accessions, and of two homologs, alpha AI-Pa present in seeds of the tepary bean (Phaseolus acutifolius) and alpha AI-Pc in seeds of the scarlet runner bean (Phaseolus coccineus), with the midgut extracts of several bruchids. The extract of the Z. subfasciatus larvae rapidly digests and inactivates alpha AI-1 and alpha AI-Pc, but not alpha AI-2 or alpha AI-Pa. The digestion is caused by a serine protease. A single proteolytic cleavage in the beta subunit of alpha AI-1 occurs at the active site of the protein. When degradation is prevented, alpha AI-1 and alpha AI-Pc do not inhibit the alpha-amylase of Z. subfasciatus, although they are effective against the alpha-amylase of C. chinensis. Alpha AI-2 and alpha AI-Pa, on the other hand, do inhibit the alpha-amylase of Z. subfasciatus, suggesting that they are good candidates for genetic engineering to achieve resistance to Z. subfasciatus. PMID:8787024

  18. Cytotoxic properties of salivary oxidants.

    PubMed

    Grisham, M B; Ryan, E M

    1990-01-01

    Salivary peroxidase and to a lesser extent myeloperoxidase are present in significant concentrations in saliva and catalyze the oxidation of thiocyanate anion (SCN-) by H2O2 to yield the potent oxidants hypothiocyanous acid (HOSCN) and its conjugate base hypothiocyanite anion (OSCN-). The objective of this study was to characterize the cytotoxic potential of peroxidase-generated HOSCN/OSCN- toward human erythrocytes. We found that HOSCN/OSCN- (0.25 mM) generated by the peroxidase-H2O2-SCN- system caused significant hemolysis at pH 6.0 but not at pH 6.5, 7.0, or 7.4. Erythrocyte hemoglobin (OxyHb) was oxidized to methemoglobin (MetHb) at all pH values tested; however, the rate of MetHb formation was dramatically increased at low pH and was not affected by inosine hexaphosphate, suggesting that hemoglobin was oxidized primarily by HOSCN. Concurrent with oxidation of hemoglobin (Hb), there was a pH-dependent consumption of HOSCN/OSCN- with more of the oxidant consumed at pH 6.0 compared with pH 6.5, 7.0, or 7.4. The enhanced oxidation of Hb at acidic pH was not due simply to increased membrane permeability by the uncharged species (HOSCN), since both erythrocyte lysate Hb and purified Hb were oxidized to the same extent at low pH as were intact erythrocytes. It is concluded that both OSCN- and HOSCN enter human erythrocytes where the protonated oxidant (HOSCN) mediates hemolysis and oxidizes OxyHb to MetHb, whereas both HOSCN and OSCN- oxidize glutathione (GSH). These data suggest that the extracellular pH may play an important role in modulating the cytotoxic properties of salivary oxidants. PMID:2154109

  19. Restoring the function of salivary glands.

    PubMed

    Kagami, H; Wang, S; Hai, B

    2008-01-01

    Salivary gland destruction occurs as a result of various pathological conditions such as radiation therapy for head and neck cancer and Sjögren's syndrome. As saliva possesses self-cleaning and antibacterial capability, hyposalivation is known to deteriorate dental caries and periodontal disease. Furthermore, hyposalivation causes mastication and swallowing problems, burning sensation of the mouth and dysgeusia. Currently available treatments for dry mouth are prescription for artificial saliva, moisturizers and medications which induce salivation from the residual tissue. Unfortunately, these treatments cannot restore the acini functions. This review focuses on various efforts to restore the function of damaged salivary gland. First, the possibility of salivary gland regeneration and tissue engineering is discussed with reference to stem cells, growth factors and scaffold materials. Second, the current status of gene transfer to salivary glands is discussed. PMID:18173444

  20. Toward salivary gland stem cell regeneration.

    PubMed

    Stiubea-Cohen, Raluca; David, Ran; Neumann, Yoav; Palmon, Aaron; Aframian, Doron

    2013-07-01

    Head and neck cancer is the sixth most common cancer worldwide, resulting in ~ 640,000 cases. Most of these patients have irreversible damage to their salivary glands due to irradiation therapy, which typically leads to significant decrease in quality of life. In the last 2 decades, several strategies have been suggested to overcome this problem; however, no biologically based treatments are available. In the past few years, the authors of the present article and other researchers have focused on a new strategy of re-implantation of autologous salivary gland cells into the residual irradiated salivary glands. This article reviews the current prospective of the irradiation-induced salivary gland impairment mechanisms and the envisioned therapeutic modalities based on stem cell therapy. PMID:24568246

  1. Treatment Option Overview (Salivary Gland Cancer)

    MedlinePLUS

    ... treat some salivary gland tumors . These include: Fast neutron radiation therapy : Fast neutron radiation therapy is a type of high-energy ... radiation therapy machine aims tiny, invisible particles, called neutrons, at the cancer cells to kill them. Fast ...

  2. [Value of echography in salivary pathology].

    PubMed

    Katz, P

    1991-05-01

    Major advances in echography techniques over the past few years include the development of simple high performance units adapted to investigating the salivary glands. This non-traumatic, painless and inexpensive technique offers a means of rapid examination and is fully complementary to X-ray and xero and sialographic imagery, particularly in tumor pathology. The author gives a review of normal images then presents different lesions of the salivary gland. PMID:1880766

  3. [Are amylases in bakery products and flour potential food allergens?].

    PubMed

    Baur, X; Sander, I; Jansen, A; Czuppon, A B

    1994-05-21

    The enzyme alpha-amylase from the mould Aspergillus oryzae (Asp o II) routinely used for the production of bread, cakes and pastries has in recent years been identified as an inhalative allergen for occupational diseases (bakers' asthma). It is doubtful whether this amylase in the final product, i.e. after the baking procedure, can still be regarded as an allergen. To clarify this question, detailed case histories on 138 subjects were recorded (98 allergics, 20 patients suffering form chronic intestinal diseases, 20 healthy controls). The clinical examinations included prick skin test and IgE antibody determination using one of the customary enzyme preparations. EAST showed a few of these 138 bread consumers to be weakly sensitized to the enzyme. One of the subjects displayed a significant reaction to alpha-amylase heated to 200 degrees C. As expected, eleven bakers sensitized to alpha-amylase by inhaling it in the workplace (positive prick test, positive case history) predominantly exhibited specific IgE antibodies to the native enzyme. Apart from one weakly positive finding, heated alpha-amylase yielded negative results in this collective. Baking conditions vary widely, especially with regard to single components, temperature and duration. Thus, further investigations as to residual allergenicity or the feasible occurrence of new antigenic determinants during the production of bread, cake and pastries are required. 27% of bakers examined and 9% of atopics showed antibodies to a flour inherent enzyme, a beta-amylase. On the whole, the selected conditions hinted at a weakly sensitizing potential inherent in baking flour and in added amylase. PMID:8209207

  4. Increased SGLT1 expression in salivary gland ductal cells correlates with hyposalivation in diabetic and hypertensive rats

    PubMed Central

    2013-01-01

    Background Oral health complications in diabetes and hypertension include decreased salivary secretion. The sodium-glucose cotransporter 1 (SGLT1) protein, which transports 1 glucose/2 Na+/264 H2O molecules, is described in salivary glands. We hypothesized that changes in SGLT1 expression in the luminal membrane of ductal cell may be related to an altered salivary flow. Findings By immunohistochemistry, we investigated SGLT1 expression in ductal cells of parotid and submandibular glands from Wistar Kyoto rats (WKY), diabetic WKY (WKY-D), spontaneously hypertensive rats (SHR) and diabetic SHR (SHR-D), as well as in parotid glands from WKY subjected to sympathetic stimulation, with or without previous propranolol blockade. Diabetes and hypertension decreased the salivary secretion and increased SGLT1 expression in the luminal membrane of ductal cells, and their association exacerbated the regulations observed. After 30 min of sympathetic stimulation, SGLT1 increased in the luminal membrane of ductal cells, and that was blocked by previous injection of propranolol. Conclusions SGLT1 expression increases in the luminal membrane of salivary gland ductal cells and the salivary flow decreases in diabetic and hypertensive rats, which may be related to sympathetic activity. This study highlights the water transporter role of SGLT1 in salivary glands, which, by increasing ductal water reabsorption, may explain the hyposalivation of diabetic and hypertensive subjects. PMID:24499577

  5. Amylase production by Preussia minima, a fungus of endophytic origin: optimization of fermentation conditions and analysis of fungal secretome by LC-MS

    PubMed Central

    2014-01-01

    Background Environmental screening programs are used to find new enzymes that may be utilized in large-scale industrial processes. Among microbial sources of new enzymes, the rationale for screening fungal endophytes as a potential source of such enzymes relates to the hypothesised mutualistic relationship between the endophyte and its host plant. There is a need for new microbial amylases that are active at low temperature and alkaline conditions as these would find industrial applications as detergents. Results An ?-amylase produced by Preussia minima, isolated from the Australian native plant, Eremophilia longifolia, was purified to homogeneity through fractional acetone precipitation and Sephadex G-200 gel filtration, followed by DEAE-Sepharose ion exchange chromatography. The purified ?-amylase showed a molecular mass of 70 kDa which was confirmed by zymography. Temperature and pH optima were 25°C and pH 9, respectively. The enzyme was activated and stabilized mainly by the metal ions manganese and calcium. Enzyme activity was also studied using different carbon and nitrogen sources. It was observed that enzyme activity was highest (138 U/mg) with starch as the carbon source and L-asparagine as the nitrogen source. Bioreactor studies showed that enzyme activity was comparable to that obtained in shaker cultures, which encourages scale-up fermentation for enzyme production. Following in-gel digestion of the purified protein by trypsin, a 9-mer peptide was sequenced and analysed by LC-ESI-MS/MS. The partial amino acid sequence of the purified enzyme presented similarity to ?-amylase from Magnaporthe oryzae. Conclusions The findings of the present study indicate that the purified ?-amylase exhibits a number of promising properties that make it a strong candidate for application in the detergent industry. To our knowledge, this is the first amylase isolated from a Preussia minima strain of endophytic origin. PMID:24602289

  6. Extracellular amylases of starch-fermenting yeast: pH effect on export and residence time in the periplasm

    Microsoft Academic Search

    G. B. Calleja; Susan R. Levy-Rick; Anwar Nasim; C. V. Lusena

    1987-01-01

    Aerobic cultures of S. alluvius in Wickerham's yeast-nitrogen-base medium with starch as sole carbon source become strongly acidic and contain no detectable extra-cellular amylolytic activity during stationary phase, when the activity in buffered cultures is maximal. The extracellular amylases are irreversibly inactivated at the low pH value (less than 3.5) attained by the cultures. When adequately buffered, the medium yields

  7. Group A Streptococcus Adheres to Pharyngeal Epithelial Cells with Salivary Proline-rich Proteins via GrpE Chaperone Protein*

    PubMed Central

    Murakami, Jumpei; Terao, Yutaka; Morisaki, Ichijiro; Hamada, Shigeyuki; Kawabata, Shigetada

    2012-01-01

    Group A Streptococcus pyogenes (GAS) is an important human pathogen that frequently causes pharyngitis. GAS organisms can adhere to and invade pharyngeal epithelial cells, which are overlaid by salivary components. However, the role of salivary components in GAS adhesion to pharyngeal cells has not been reported precisely. We collected human saliva and purified various salivary components, including proline-rich protein (PRP), statherin, and amylase, and performed invasion assays. The GAS-HEp-2 association ratio (invasion/adhesion ratio) and invasion ratio of GAS were increased significantly with whole human saliva and PRP, while the anti-PRP antibody inhibited the latter. GAS strain NY-5, which lacks M and F proteins on the cell surface, was promoted to cohere with HEp-2 cells by whole human saliva and PRP. The 28-kDa protein of GAS bound to PRP and was identified as GrpE, a chaperone protein, whereas the N-terminal of GrpE was found to bind to PRP. A GrpE-deficient mutant of GAS strain B514Sm, TR-45, exhibited a reduced ability to adhere to and invade HEp-2 cells. Microscopic observations showed the GrpE was mainly expressed on the surface of the cell division site of GAS. Furthermore, GrpE-deficient mutants of GAS and Streptococcus pneumoniae showed an elongated morphology as compared with the wild type. Taken together, this is the first study to show an interaction between salivary PRP and GAS GrpE, which plays an important role in GAS infection on the pharynx, whereas the expression of GrpE on the surface of GAS helps to maintain morphology. PMID:22566698

  8. Variability in the production of amylase by three isolates of Myrothecium roridum

    Microsoft Academic Search

    P. B. Reddy; S. M. Reddy

    1989-01-01

    Amylase production by three isolates ofMyrothecium roridum under different cultural conditions was studied. Starch followed by dextrin induced maximum amylase production (dextrinizing\\u000a and saccharifying) by all the three isolates. Glucose was a poor substrate for the production of amylases. Bitter gourd isolate\\u000a was a comparatively more efficient producer of amylases than the other two isolates. Addition of glucose to the

  9. Nucleotide sequence of the alpha-amylase gene (ALP1) in the yeast Saccharomycopsis fibuligera.

    PubMed

    Itoh, T; Yamashita, I; Fukui, S

    1987-07-27

    The complete nucleotide sequence of the secretable alpha-amylase gene ALP1 from the yeast Saccharomycopsis fibuligera has been determined. The ALP1 DNA hybridized to a polyadenylated RNA of 2.0 kilobases. A single open reading frame encodes a 494-amino acid protein which is highly homologous with alpha-amylase (Taka-amylase) of a fungus Aspergillus oryzae. PMID:3497057

  10. Functional conservation of a glucose-repressible amylase gene promoter from Drosophila virilis in Drosophila melanogaster

    Microsoft Academic Search

    Charalambos Magoulas; Ada Loverre-Chyurlia; Sumaia Abukashawa; Laure Bally-Cuff; Donal A. Hickey

    1993-01-01

    Previous studies have demonstrated that the expression of the a-amylase gene is repressed by dietary glucose in Drosophila melanogaster. Here, we show that the a-amylase gene of a distantly related species, D. virilis, is also subject to glucose repression. Moreover, the cloned amylase gene of D. virilis is shown to be glucose repressible when it is transiently expressed in D.

  11. AmyM, a Novel Maltohexaose-Forming ?-Amylase from Corallococcus sp. Strain EGB.

    PubMed

    Li, Zhoukun; Wu, Jiale; Zhang, Biying; Wang, Fei; Ye, Xianfeng; Huang, Yan; Huang, Qiang; Cui, Zhongli

    2015-03-15

    A novel ?-amylase, AmyM, was purified from the culture supernatant of Corallococcus sp. strain EGB. AmyM is a maltohexaose-forming exoamylase with an apparent molecular mass of 43 kDa. Based on the results of matrix-assisted laser desorption ionization-time of flight mass spectrometry and peptide mass fingerprinting of AmyM and by comparison to the genome sequence of Corallococcus coralloides DSM 2259, the AmyM gene was identified and cloned into Escherichia coli. amyM encodes a secretory amylase with a predicted signal peptide of 23 amino acid residues, which showed no significant identity with known and functionally verified amylases. amyM was expressed in E. coli BL21(DE3) cells with a hexahistidine tag. The signal peptide efficiently induced the secretion of mature AmyM in E. coli. Recombinant AmyM (rAmyM) was purified by Ni-nitrilotriacetic acid (NTA) affinity chromatography, with a specific activity of up to 14,000 U/mg. rAmyM was optimally active at 50°C in Tris-HCl buffer (50 mM; pH 7.0) and stable at temperatures of <50°C. rAmyM was stable over a wide range of pH values (from pH 5.0 to 10.0) and highly tolerant to high concentrations of salts, detergents, and various organic solvents. Its activity toward starch was independent of calcium ions. The Km and Vmax of recombinant AmyM for soluble starch were 6.61 mg ml(-1) and 44,301.5 ?mol min(-1) mg(-1), respectively. End product analysis showed that maltohexaose accounted for 59.4% of the maltooligosaccharides produced. These characteristics indicate that AmyM has great potential in industrial applications. PMID:25576603

  12. What's New in Salivary Gland Cancer Research and Treatment?

    MedlinePLUS

    ... Topic Additional resources for salivary gland cancer What’s new in salivary gland cancer research and treatment? Medical ... they hope to use this information to develop new treatments that work better and cause fewer side ...

  13. Original article Quantitation of histochemical staining of salivary

    E-print Network

    Paris-Sud XI, Université de

    alone. Quantitative assessment of normal salivary gland histochemistry allows com- parison with stainingOriginal article Quantitation of histochemical staining of salivary gland mucin using image were employed to complement subjective assessment of patterns of mucin staining (by periodic acid

  14. Saliva, salivary gland, and hemolymph collection from Ixodes scapularis ticks.

    PubMed

    Patton, Toni G; Dietrich, Gabrielle; Brandt, Kevin; Dolan, Marc C; Piesman, Joseph; Gilmore, Robert D

    2012-01-01

    Ticks are found worldwide and afflict humans with many tick-borne illnesses. Ticks are vectors for pathogens that cause Lyme disease and tick-borne relapsing fever (Borrelia spp.), Rocky Mountain Spotted fever (Rickettsia rickettsii), ehrlichiosis (Ehrlichia chaffeensis and E. equi), anaplasmosis (Anaplasma phagocytophilum), encephalitis (tick-borne encephalitis virus), babesiosis (Babesia spp.), Colorado tick fever (Coltivirus), and tularemia (Francisella tularensis) (1-8). To be properly transmitted into the host these infectious agents differentially regulate gene expression, interact with tick proteins, and migrate through the tick (3,9-13). For example, the Lyme disease agent, Borrelia burgdorferi, adapts through differential gene expression to the feast and famine stages of the tick's enzootic cycle (14,15). Furthermore, as an Ixodes tick consumes a bloodmeal Borrelia replicate and migrate from the midgut into the hemocoel, where they travel to the salivary glands and are transmitted into the host with the expelled saliva (9,16-19). As a tick feeds the host typically responds with a strong hemostatic and innate immune response (11,13,20-22). Despite these host responses, I. scapularis can feed for several days because tick saliva contains proteins that are immunomodulatory, lytic agents, anticoagulants, and fibrinolysins to aid the tick feeding (3,11,20,21,23). The immunomodulatory activities possessed by tick saliva or salivary gland extract (SGE) facilitate transmission, proliferation, and dissemination of numerous tick-borne pathogens (3,20,24-27). To further understand how tick-borne infectious agents cause disease it is essential to dissect actively feeding ticks and collect tick saliva. This video protocol demonstrates dissection techniques for the collection of hemolymph and the removal of salivary glands from actively feeding I. scapularis nymphs after 48 and 72 hours post mouse placement. We also demonstrate saliva collection from an adult female I. scapularis tick. PMID:22371172

  15. Salivary pH: A diagnostic biomarker

    PubMed Central

    Baliga, Sharmila; Muglikar, Sangeeta; Kale, Rahul

    2013-01-01

    Objectives: Saliva contains a variety of host defense factors. It influences calculus formation and periodontal disease. Different studies have been done to find exact correlation of salivary biomarkers with periodontal disease. With a multitude of biomarkers and complexities in their determination, the salivary pH may be tried to be used as a quick chairside test. The aim of this study was to analyze the pH of saliva and determine its relevance to the severity of periodontal disease. Study Design: The study population consisted of 300 patients. They were divided into three groups of 100 patients each: Group A had clinically healthy gingiva, Group B who had generalized chronic gingivitis and Group C who had generalized chronic periodontitis. The randomized unstimulated saliva from each patient was collected and pH was tested. Data was analyzed statistically using analysis of variance technique. Results: The salivary pH was more alkaline for patients with generalized chronic gingivitis as compared with the control group (P = 0.001) whereas patients with generalized chronic periodontitis had more acidic pH as compared with the control group (P = 0.001). Conclusion: These results indicate a significant change in the pH depending on the severity of the periodontal condition. The salivary pH shows significant changes and thus relevance to the severity of periodontal disease. Salivary pH may thus be used as a quick chairside diagnostic biomarker. PMID:24174725

  16. Immobilization of ?-amylase onto poly(glycidyl methacrylate) grafted electrospun fibers by ATRP.

    PubMed

    Oktay, Burcu; Demir, Serap; Kayaman-Apohan, Nilhan

    2015-05-01

    In this study, novel ?-amylase immobilized poly(vinyl alcohol) (PVA) nanofibers were prepared. The PVA nanofiber surfaces were functionalized with 2-bromoisobutyryl bromide (BiBBr) and followed by surface initiated atom transfer radical polymerization (SI-ATRP) of glycidyl methacrylate (GMA). The morphology of the poly(glycidyl methacrylate) (PGMA) grafted PVA nanofibers was characterized by scanning electron microscopy (SEM). Also PGMA brushes were confirmed by X-ray photo electron microscopy (XPS). ?-Amylase was immobilized in a one step process onto the PGMA grafted PVA nanofiber. The characteristic properties of the immobilized and free enzymes were examined. The thermal stability of the enzyme was improved and showed maximum activity at 37°C by immobilization. pH values of the maximum activity of the free and immobilized enzymes were also found at 6.0 and 6.5, respectively. Free enzyme lost its activity completely within 15days. The immobilized enzyme lost only 23.8% of its activity within 30days. PMID:25746284

  17. Establishment of immortal multipotent rat salivary progenitor cell line toward salivary gland regeneration.

    PubMed

    Yaniv, Adi; Neumann, Yoav; David, Ran; Stiubea-Cohen, Raluca; Orbach, Yoav; Lang, Stephan; Rotter, Nicole; Dvir-Ginzberg, Mona; Aframian, Doron J; Palmon, Aaron

    2011-01-01

    Adult salivary gland stem cells are promising candidates for cell therapy and tissue regeneration in cases of irreversible damage to salivary glands in head and neck cancer patients undergoing irradiation therapy. At present, the major restriction in handling such cells is their relatively limited life span during in vitro cultivation, resulting in an inadequate experimental platform to explore the salivary gland-originated stem cells as candidates for future clinical application in therapy. We established a spontaneous immortal integrin ?6?1-expressing cell line of adult salivary progenitor cells from rats (rat salivary clone [RSC]) and investigated their ability to sustain cellular properties. This line was able to propagate for more than 400 doublings without loss of differentiation potential. RSC could differentiate in vitro to both acinar- and ductal-like structures and could be further manipulated upon culturing on a 3D scaffolds with different media supplements. Moreover, RSC expressed salivary-specific mRNAs and proteins as well as epithelial stem cell markers, and upon differentiation process their expression was changed. These results suggest RSC as a good model for further studies exploring cellular senescence, differentiation, and in vitro tissue engineering features as a crucial step toward reengineering irradiation-impaired salivary glands. PMID:20673137

  18. Cloning and expression of human salivary-gland kallikrein in Escherichia coli.

    PubMed Central

    Angermann, A; Bergmann, C; Appelhans, H

    1989-01-01

    CDNA clones for human kallikrein have been identified in a cDNA library constructed from mRNA of human salivary gland. The entire coding sequence for preprokallikrein and for the 5'- and 3'-untranslated regions were isolated by using a mixture of oligonucleotides corresponding to amino acids 51-56 of human urinary kallikrein and one oligonucleotide corresponding to amino acids 233-238 of human pancreatic kallikrein. The DNA sequence proved that, with the exception of two amino acid exchanges, kallikrein of the human salivary gland is identical with pancreatic kallikrein. Salivary gland and renal kallikrein was expressed in Escherichia coli from plasmid pKK223-3 under the control of the tac promoter. The protein was identified by Western-blot analysis and by demonstration of its specific proteolytic activity. Images Fig. 1. Fig. 4. PMID:2686621

  19. Salivary proteins of Russian wheat aphid (Hempitera: Aphididae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salivary secretions play critical roles in aphid – host plant interactions and are responsible for damage associated with aphid feeding. The objectives of this study were to evaluate aspects of salivation and the salivary constituents of Diuraphis noxia (Hemiptera: Aphididae). Salivary proteins we...

  20. Optical recognition of salivary proteins by use of molecularly imprinted poly(ethylene-co-vinyl alcohol)/quantum dot composite nanoparticles.

    PubMed

    Lee, Mei-Hwa; Chen, Yun-Chao; Ho, Min-Hsien; Lin, Hung-Yin

    2010-06-01

    Molecularly imprinted polymers (MIPs) have long been studied for applications in biomolecule recognition and binding; compared with natural antibodies, they may offer advantages in cost and stability. We report on the development of MIPs that "self-report" concentrations of bound analytes via fluorescence changes in embedded quantum dots (QDots). Composite QDot/MIPs were prepared using phase inversion of poly(ethylene-co-vinyl alcohol) (EVAL) solutions with various ethylene mole ratios in the presence of salivary target molecules (e.g. amylase, lipase, and lysozyme). These major protein components of saliva have been implicated as possible biomarkers for pancreatic cancer. The optimum (highest imprinting effectiveness) ethylene mole ratios of the commercially available EVALs were found to be 32, 38, and 44 mol% for the imprinting of amylase, lipase, and lysozyme, respectively. QD fluorescence quenching was observed on binding of analytes to composite MIPs in a concentration-dependent manner, and was used to construct calibration curves. Finally, the composite MIP particles were used for the quantitative detection of amylase, lipase, and lysozyme in real samples (saliva) and compared with a commercial Architect ci 8200 chemical analysis system. PMID:20349227

  1. Secretion to the growth medium of an ?-amylase by Escherichia coli clones carrying a Bacillus laterosporus gene

    Microsoft Academic Search

    H. K. Manonmani; A. A. M. Kunhi

    1999-01-01

    a-Amylase gene from Bacillus laterosporus P3 was cloned and expressed in Escherichia coli HB101 and DH5a. Up to 92% of the cloned gene product was secreted into the medium by the recombinant E. coli. The recombinant crude enzyme showed improved functionality in terms of activity at a wider pH range and at higher temperature, as compared to the crude enzyme

  2. NETRIN and SLIT guide salivary gland migration.

    PubMed

    Kolesnikov, Tereza; Beckendorf, Steven K

    2005-08-01

    Directed migration is pivotal for the proper placement and function of nearly all organs. The majority of known guidance molecules involved in directed migration have been identified from studies of migrating axons during nervous system development. Here, we show that at least two of these axon guidance molecules, NETRIN and SLIT, act through their canonical receptors, to guide Drosophila embryonic salivary glands. NETRIN serves as a chemo-attractant while SLIT functions antagonistically to NETRIN as a chemo-repellent during salivary gland migration. CNS midline expression of both NETRIN and SLIT directs the glands to move unswervingly parallel to the CNS. NETRIN expression is also required in the visceral mesoderm, along which the glands move during their migration. We propose that analogous to axon guidance, a balance between chemo-attractants and chemo-repellents is required for the proper migratory path of the developing salivary glands. PMID:15950216

  3. On the mechanism of salivary gland radiosensitivity

    SciTech Connect

    Konings, Antonius W.T. [Department of Radiation and Stress Cell Biology, University of Groningen, Groningen (Netherlands)]. E-mail: a.w.t.konings@med.rug.nl; Coppes, Rob P. [Department of Radiation and Stress Cell Biology, University of Groningen, Groningen (Netherlands); Department of Radiation Oncology, University Hospital Groningen, Groningen (Netherlands); Vissink, Arjan [Department of Oral and Maxillofacial Surgery, University Hospital, Groningen (Netherlands)

    2005-07-15

    Purpose: To contribute to the understanding of the enigmatic radiosensitivity of the salivary glands by analysis of appropriate literature, especially with respect to mechanisms of action of early radiation damage, and to supply information on the possibilities of amelioration of radiation damage to the salivary glands after radiotherapy of head-and-neck cancer. Methods and Materials: Selected published data on the mechanism of salivary gland radiosensitivity and radioprotection were studied and analyzed. Results: From a classical point of view, the salivary glands should not respond as rapidly to radiation as they appear to do. Next to the suggestion of massive apoptosis, the leakage of granules and subsequent lysis of acinar cells was suggested to be responsible for the acute radiation-induced function loss of the salivary glands. The main problem with these hypotheses is that recently performed assays show no cell loss during the first days after irradiation, while saliva flow is dramatically diminished. The water secretion is selectively hampered during the first days after single-dose irradiation. Literature is discussed that shows that the compromised cells suffer selective radiation damage to the plasma membrane, disturbing signal transduction primarily affecting watery secretion. Although the cellular composition of the submandibular gland and the parotid gland are different, the damage response is very alike. The acute radiation-induced function loss in both salivary glands can be ameliorated by prophylactic treatment with specific receptor agonists. Conclusions: The most probable mechanism of action, explaining the enigmatic high radiosensitivity for early effects, is selective radiation damage to the plasma membrane of the secretory cells, disturbing muscarinic receptor stimulated watery secretion. Later damage is mainly due to classical mitotic cell death of progenitor cells, leading to a hampered replacement capacity of the gland for secretory cells, but is also caused by damage to the extracellular environment, preventing proper cell functioning.

  4. Clinical and immunological responses to occupational exposure to alpha-amylase in the baking industry.

    PubMed Central

    Brisman, J; Belin, L

    1991-01-01

    alpha-Amylase is a starch cleaving enzyme often used in the baking industry as a flour additive. It is usually of fungal origin, produced by Aspergillus oryzae. One previous report has shown IgE antibodies and positive skin prick test against alpha-amylase in asthmatic bakers. This paper describes four alpha-amylase sensitised index cases with occupational asthma or rhinitis and the results of a cross sectional study of 20 workers from the same factory who were also exposed to alpha-amylase powder. Air sampling detected airborne alpha-amylase at a concentration of 0.03 mg/m3. Significantly more work related symptoms such as rhinitis and dermatitis were found among the alpha-amylase exposed workers compared with referents. A skin prick test to alpha-amylase was positive in 30% (6/20) of the exposed workers. Most of the persons showing a positive skin prick test had work related symptoms and were also skin prick test positive to common allergens. Nasal challenge tests with amylase were performed in selected cases and validated three cases of alpha-amylase induced rhinitis. Two non-symptomatic workers had precipitins to alpha-amylase. Specific IgG antibodies were shown by two further serological techniques. The nature and relevance of these antibodies are currently being studied. It is concluded that alpha-amylase powder is a potent occupational sensitiser. Precautions should be taken when handling this allergenic enzyme. PMID:1832939

  5. Biochemical characterization of alpha-amylase from the yeast Cryptococcus flavus.

    PubMed

    Wanderley, Kenya J; Torres, Fernando A G; Moraes, Lídia M P; Ulhoa, Cirano J

    2004-02-16

    During our screening of amylolytic microorganisms from Brazilian fruits, we isolated a yeast strain classified as Cryptococcus flavus. When grown on starch-containing medium this strain exhibited the highest amylase production after 24 h of cultivation. The extracellular amylase from C. flavus was purified from the culture broth by a single step using chromatography on a Sephacryl S-100 column. The enzyme was purified 16.14-fold with a yield of 50.21% of the total activity. The purified enzyme was a glycoprotein with an apparent molecular mass of 75 and 84.5 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration, respectively. The enzyme lost approximately 50% of the molecular mass after treatment with glycosidases. The major end products of starch, amylose, amylopectin, pullulan and glycogen were maltose and maltotriose. The K(m) value for the pure enzyme was 0.056 mg ml(-1) with soluble starch as the substrate. Enzyme activity was optimal at pH 5.5 and 50 degrees C. The enzyme retained 90% of the activity after incubation at 50 degrees C for 60 min and was inhibited by Cu(2+), Fe(2+) and Hg(2+). PMID:14987760

  6. Cephalotoxin: the Crab-paralysing Agent of the Posterior Salivary Glands of Cephalopods

    Microsoft Academic Search

    F. Ghiretti

    1959-01-01

    THE presence of a substance, toxic to crabs, in the posterior salivary glands of cephalopods (Giftdrusen of the German authors) has been known for a long time1. When a drop of saliva collected from Octopus is injected into a crab, a sequence of reactions is observed followed by complete paralysis of the animal2. Active substances have been extracted from the

  7. Highly thermostable and alkaline ?-amylase from a halotolerant-alkaliphilic Bacillus sp. AB68

    PubMed Central

    Aygan, Ashabil; Arikan, Burhan; Korkmaz, Hatice; Dinçer, Sadik; Çolak, Ömer

    2008-01-01

    An alkaliphilic and highly thermostable ?-amylase producing Bacillus sp. was isolated from Van soda lake. Enzyme synthesis occurred at temperatures between 25°C and 40°C. Analysis of the enzyme by SDS-PAGE revealed a single band which was estimated to be 66 kDa. The enzyme was active in a broad temperature range, between 20°C and 90°C, with an optimum at 50°C; and maximum activity was at pH 10.5. The enzyme was almost completely stable up to 80°C with a remaining activity over 90% after 30 min pre-incubation. Thermostability was not increased in the presence of Ca2+. An average of 75% and 60°C of remaining activity was observed when the enzyme was incubated between pH 5 and 9 for 1 h and for 2 h, respectively. The activity of the enzyme was inhibited by SDS and EDTA by 38% and 34%, respectively. PMID:24031264

  8. Regulation of amylase release from dispersed pancreatic acinar cells.

    PubMed Central

    Gardner, J D; Jackson, M J

    1977-01-01

    1. A study has been made of factors influencing release of amylase from dispersed pancreatic acinar cells. 2. In the basal, unstimulated, condition cells released 2-3% of the total amylase present in 30 min. 3. The rate of amylase release was stimulated 50-70% by C-terminal octapeptide of cholecystokinin (CCK-OP, maximally effective concentration, 3 X 10(-10) M); carbamylcholine (maximally effective concentration, 10(-5 M); secretin (maximally effective concentration greater than 10(-6) M); vasoactive intestinal peptide (VIP, maximally effective concentration, 10(-8) M); and adenosine 3':5' monophosphate (cyclic AMP) and guanosine 3':5' monophosphate (cyclic GMP) as well as their dibutyryl derivatives (maximally effective concentrations, 10(-3) M). 4. The responses to CCK-OP or carbamylcholine were potentiated by secretin, VIP or dibutyryl cyclic AMP. 5. The responses to secretin or VIP were potentiated by CCK-OP, carbamylcholine, or dibutyryl cyclic GMP. 6. There appear to be two pathways for the regulation of amylase release from pancreatic acinar cells: one pathway can be stimulated by cholecystokinin or cholinergic agonists, and the response to these stimuli is mediated by cyclic GMP; the other pathway can be stimulated by secretin or VIP, and the response to these stimuli is mediated by cyclic AMP. PMID:198531

  9. Enzymatic properties and primary structures of two ?-amylase isozymes from the Pacific abalone Haliotis discus hannai.

    PubMed

    Kumagai, Yuya; Satoh, Takuya; Inoue, Akira; Ojima, Takao

    2013-02-01

    Two ?-amylase (EC 3.2.1.1) isozymes, HdAmy58 and HdAmy82, with approximate molecular masses of 58 kDa and 82 kDa, respectively, were isolated from the digestive fluid of the Pacific abalone Haliotis discus hannai. Optimal temperatures and pHs for HdAmy58 and HdAmy82 were at 30 °C and 6.7, and 30 °C and 6.1, respectively. Both enzymes similarly degraded starch, glycogen, and maltooligosaccharides larger than maltotriose producing maltose and maltotriose as the major degradation products. However, the activity toward maltotetraose was appreciably higher in HdAmy82 than HdAmy58. cDNAs encoding HdAmy58 and HdAmy82 were cloned and the amino-acid sequences of 511 and 694 residues for HdAmy58 and HdAmy82, respectively, were deduced. The putative catalytic domains of HdAmy58 and HdAmy82 were located in the 17-511th and 19-500th amino-acid regions, respectively, and they showed approximately 50% amino-acid identity to each other. These sequences also showed 62-99% amino-acid identity to the catalytic domains of known ?-amylases that belong to glycoside-hydrolase-family 13. The difference in the molecular masses between HdAmy58 and HdAmy82 was ascribed to the extension of approximately 190 residues in the C-terminus of HdAmy82. This extended region showed 41-63% amino-acid identity with the ancillary domains of several ?-amylases previously reported. PMID:23142215

  10. Maltose-forming ?-amylase from the hyperthermophilic archaeon Pyrococcus sp. ST04.

    PubMed

    Jung, Jong-Hyun; Seo, Dong-Ho; Holden, James F; Park, Cheon-Seok

    2014-03-01

    The deduced amino acid sequence from a gene of the hyperthermophilic archaeon Pyrococcus sp. ST04 (Py04_0872) contained a conserved glycoside hydrolase family 57 (GH57) motif, but showed <13% sequence identity with other known Pyrococcus GH57 enzymes, such as 4-?-glucanotransferase (EC 2.4.1.25), amylopullulanase (EC 3.2.1.41), and branching enzyme (EC 2.4.1.18). This gene was cloned and expressed in Escherichia coli, and the recombinant product (Pyrococcus sp. ST04 maltose-forming ?-amylase, PSMA) was a novel 70-kDa maltose-forming ?-amylase. PSMA only recognized maltose (G2) units with ?-1,4 and ?-1,6 linkages in polysaccharides (e.g., starch, amylopectin, and glycogen) and hydrolyzed pullulan very poorly. G2 was the primary end product of hydrolysis. Branched cyclodextrin (CD) was only hydrolyzed along its branched maltooligosaccharides. 6-O-glucosyl-?-cyclodextrin (G1-?-CD) and ?-cyclodextrin (?-CD) were resistant to PSMA suggesting that PSMA is an exo-type glucan hydrolase with ?-1,4- and ?-1,6-glucan hydrolytic activities. The half-saturation value (Km) for the ?-1,4 linkage of maltotriose (G3) was 8.4 mM while that of the ?-1,6 linkage of 6-O-maltosyl-?-cyclodextrin (G2-?-CD) was 0.3 mM. The kcat values were 381.0 min(-1) for G3 and 1,545.0 min(-1) for G2-?-CD. The enzyme was inhibited competitively by the reaction product G2, and the Ki constant was 0.7 mM. PSMA bridges the gap between amylases that hydrolyze larger maltodextrins and ?-glucosidase that feeds G2 into glycolysis by hydrolyzing smaller glucans into G2 units. PMID:23884203

  11. A review: Immunological markers for malignant salivary gland tumors

    PubMed Central

    Namboodiripad, P.C. Anila

    2014-01-01

    Salivary gland cancers are rare. Around 8 out of 10 salivary gland tumors (80%) are in the parotid. Just fewer than 2 out of 10 salivary gland cancers develop in the other two salivary glands – the submandibular or sublingual glands. Fewer than 1 in 10 cancers start in the minor salivary glands. There are many different types of salivary gland cancers. The most common is mucoepidermoid carcinoma (MEC). Just over 3 out of 10 (25–35%) salivary gland cancers (SGT, SGC) are of this type. The others include adenoid cystic carcinoma (ACC), acinic cell carcinoma, carcinoma ex-pleomorphic adenoma (Ca-PA), polymorphous low grade adenocarcinoma (PLGA) and some newly discovered salivary gland tumors. Because of the infrequency of salivary gland tumors and their complex histopathological diagnosis, it is difficult to exactly predict their clinical course by means of its recurrence, malignant progression or metastasis. Salivary gland tumors always pose problems in diagnosis. This review provides an insight into the recent concepts and immunohistochemical markers to diagnose the malignant salivary gland tumors (SGT), thus guiding the Ear, Nose and Throat specialists, Oral and Maxillofacial Surgeons, General Pathologists and other medical and dental specialists thereby enabling them to make correct diagnosis and provide the appropriate treatment. PMID:25737930

  12. Identification of the three major coeliac immunoreactive proteins and one alpha-amylase inhibitor from oat endosperm.

    PubMed

    Rocher, A; Colilla, F; Ortiz, M L; Mendez, E

    1992-09-21

    Six chloroform/methanol-soluble proteins from oat endosperm (Avena sativa) have been isolated and characterized by a purification procedure based on extraction with volatile solvents, followed by reversed-phase high performance liquid chromatography. Three of these proteins, with an assessed molecular weight of 25,000, 27,000 and 32,000 Da, respectively, have been identified by immunoblotting using coeliac sera, as the major coeliac serum IgA-binding components of oat endosperm. The N-terminal amino acid sequence of these proteins indicates that they correspond to alpha 2, gamma 4, and gamma 3 avenins, respectively. We have tentatively named them 'coeliac immunoreactive proteins'. Another chloroform/methanol oat component shows weak alpha-amylase inhibitory activity and exhibits strong homology (60% identity) at the N-terminus with the alpha-amylase inhibitor from ragi (Eleusine coracana). PMID:1526282

  13. A comparative study on the inhibitory effects of different parts and chemical constituents of pomegranate on ?-amylase and ?-glucosidase.

    PubMed

    Kam, Antony; Li, Kong M; Razmovski-Naumovski, Valentina; Nammi, Srinivas; Shi, Jeffrey; Chan, Kelvin; Li, George Q

    2013-11-01

    Pomegranate has been documented for the management of diabetes in Unani and Chinese medicine. This study compared the effects of the extracts of different pomegranate parts, including juice, peels, seeds and flowers, on carbohydrate digestive enzymes (?-amylase and ?-glucosidase) in vitro. The methanolic flower extract inhibited ?-amylase and ?-glucosidase, while the methanolic peel extract inhibited ?-glucosidase selectively. The most active flower extract was subjected to water-ethyl acetate partition. The ethyl acetate fraction was more potent than the water fraction in inhibiting both enzymes. Gallic acid and ellagic acid also showed selective inhibition against ?-glucosidase, and their presence in the ethyl acetate fraction was confirmed by HPLC-DAD and HPLC-HESI-MS. Our findings suggest that the inhibition of carbohydrate digestive enzymes and their phenolic content may contribute to the anti-hyperglycaemic effects of pomegranate flower and peel, and support their claims in diabetes. PMID:23280757

  14. Molecular cues for development and regeneration of salivary glands.

    PubMed

    Liu, Fei; Wang, Songlin

    2014-03-01

    The hypofunction of salivary glands caused by Sjögren's Syndrome or radiotherapy for head and neck cancer significantly compromises the quality of life of millions patients. Currently no curative treatment is available for the irreversible hyposalivation, whereas regenerative strategies targeting salivary stem/progenitor cells are promising. However, the success of these strategies is constrained by the lack of insights on the molecular cues of salivary gland regeneration. Recent advances in the molecular controls of salivary gland morphogenesis provided valuable clues for identifying potential regenerative cues. A complicated network of signaling molecules between epithelia, mesenchyme, endothelia, extracellular matrix and innervating nerves orchestrate the salivary gland organogenesis. Here we discuss the roles of several cross-talking intercellular signaling pathways, i.e., FGF, Wnt, Hedgehog, Eda, Notch, Chrm1/HB-EGF and Laminin/Integrin pathways, in the development of salivary glands and their potentials to promote salivary regeneration. PMID:24189993

  15. Current therapies for xerostomia and salivary gland hypofunction associated with cancer therapies.

    PubMed

    Nieuw Amerongen, A V; Veerman, E C I

    2003-04-01

    In cancer patients, as in the general population, medication is the most common cause of xerostomia. In general, saliva flow in these patients can be stimulated by mechanical or pharmacological stimulation of the salivary glands. Painful damaged oral mucosa can be treated by softening, lubricating mouthwashes or gels. A specific group of patients are those receiving radiotherapy for malignant tumours in the head and neck region. This treatment is inevitably associated with damages to the oral tissues, including the salivary glands, resulting in salivary gland hypofunction. When residual secretory capacity is present, it is advisable to stimulate the salivary glands by mechanical or gustatory stimuli regularly in these patients as supportive oral care. Alternatively, salivary flow can be stimulated by the use of cholinergic pharmaceutical preparations, such as pilocarpine or cevimeline. After the radiation therapy is ended, a dental check-up should be done every 3 months to allow control of any incipient oral inflammation and dental decay. For daily use, a special dentifrice (e.g. children's toothpaste) is recommended, since the taste of a regular dentifrice may be too strong for these patients. Nocturnal oral dryness can be alleviated by spraying the oral surfaces with water, or by applying a small amount of dentifrice on the dental smooth surfaces. When stimulation of salivary secretion fails, patients can be given palliative oral care in the form of application of mouthwashes and saliva substitutes. The daily use of a mouthwash, e.g. Biotène, Oral Balance or Zendium, or one of the saliva substitutes is indicated. Different types of saliva substitutes are now commercially available, containing different polymers as thickening agents, e.g. carboxymethylcellulose (Oralube and Glandosane), polyacrylic acid, and xanthan gum (Xialine). Recent developments, which are, however, still in the experimental stage, are bio-active saliva substitutes and mouthwashes containing antimicrobial peptides to protect the oral tissues against microbial colonization and to suppress and to cure mucosal and gingival inflammation. PMID:12673460

  16. Salivary testosterone levels among Tamang and Kami males of central Nepal.

    PubMed

    Ellison, P T; Panter-Brick, C

    1996-12-01

    Salivary testosterone levels are reported for 65 Nepalese males between the ages of 15 and 48 years who were drawn from 2 different ethnic populations (Tamang and Kami) from the central highlands of Nepal. Subjects collected morning and evening saliva samples on five consecutive days in two contrasting seasons, the winter dry season and the summer monsoon season. Anthropometric indexes of acute and chronic nutritional status were also measured. Morning and evening salivary testosterone levels in the winter averaged 233 +/- 14 (SE) pmol/L and 166 +/- 8 pmol/L, respectively, for the Tamang and 249 +/- 14 pmol/L and 163 +/- 13 pmol/L, respectively, for the Kami. In the summer the corresponding values were 219 +/- 12 pmol/L and 156 +/- 8 pmol/L for the Tamang and 249 +/- 19 pmol/L and 147 +/- 12 pmol/L for the Kami. These levels are significantly lower than those reported for Western populations and close to those reported for other non-Western populations. The magnitude of diurnal variation in salivary testosterone levels and the absence of significant age variation are also comparable with observations made on other populations. Weak relationships were observed between testosterone levels and indexes of acute and chronic nutritional status in the winter only. The absence of pronounced variation in salivary testosterone levels between populations and the absence of strong associations between salivary testosterone levels and indexes of acute and chronic nutritional status contrast with the prominent ecological and interpopulation variation reported for salivary progesterone levels in women. Male gonadal function seems less sensitive to moderate energetic stress than female gonadal function, probably reflecting the fact that energy availability is less crucial to male reproductive success than to female reproductive success. Variation in testosterone level associated with chronic energetic stress may be an adaptive somatic response to avoid the maintenance costs of a large active metabolic mass with little direct impact on male fecundity. PMID:8979466

  17. Improving the thermostability of alpha-amylase by combinatorial coevolving-site saturation mutagenesis

    PubMed Central

    2012-01-01

    Background The generation of focused mutant libraries at hotspot residues is an important strategy in directed protein evolution. Existing methods, such as combinatorial active site testing and residual coupling analysis, depend primarily on the evolutionary conserved information to find the hotspot residues. Hardly any attention has been paid to another important functional and structural determinants, the functionally correlated variation information--coevolution. Results In this paper, we suggest a new method, named combinatorial coevolving-site saturation mutagenesis (CCSM), in which the functionally correlated variation sites of proteins are chosen as the hotspot sites to construct focused mutant libraries. The CCSM approach was used to improve the thermal stability of ?-amylase from Bacillus subtilis CN7 (Amy7C). The results indicate that the CCSM can identify novel beneficial mutation sites, and enhance the thermal stability of wild-type Amy7C by 8°C ( T5030), which could not be achieved with the ordinarily rational introduction of single or a double point mutation. Conclusions Our method is able to produce more thermostable mutant ?-amylases with novel beneficial mutations at new sites. It is also verified that the coevolving sites can be used as the hotspots to construct focused mutant libraries in protein engineering. This study throws new light on the active researches of the molecular coevolution. PMID:23057711

  18. ?-Amylase–Like Proteins Function as Transcription Factors in Arabidopsis, Controlling Shoot Growth and Development[C][W][OA

    PubMed Central

    Reinhold, Heike; Soyk, Sebastian; Šimková, Klára; Hostettler, Carmen; Marafino, John; Mainiero, Samantha; Vaughan, Cara K.; Monroe, Jonathan D.; Zeeman, Samuel C.

    2011-01-01

    Plants contain ?-amylase–like proteins (BAMs; enzymes usually associated with starch breakdown) present in the nucleus rather than targeted to the chloroplast. They possess BRASSINAZOLE RESISTANT1 (BZR1)-type DNA binding domains—also found in transcription factors mediating brassinosteroid (BR) responses. The two Arabidopsis thaliana BZR1-BAM proteins (BAM7 and BAM8) bind a cis-regulatory element that both contains a G box and resembles a BR-responsive element. In protoplast transactivation assays, these BZR1-BAMs activate gene expression. Structural modeling suggests that the BAM domain’s glucan binding cleft is intact, but the recombinant proteins are at least 1000 times less active than chloroplastic ?-amylases. Deregulation of BZR1-BAMs (the bam7bam8 double mutant and BAM8-overexpressing plants) causes altered leaf growth and development. Of the genes upregulated in plants overexpressing BAM8 and downregulated in bam7bam8 plants, many carry the cis-regulatory element in their promoters. Many genes that respond to BRs are inversely regulated by BZR1-BAMs. We propose a role for BZR1-BAMs in controlling plant growth and development through crosstalk with BR signaling. Furthermore, we speculate that BZR1-BAMs may transmit metabolic signals by binding a ligand in their BAM domain, although diurnal changes in the concentration of maltose, a candidate ligand produced by chloroplastic ?-amylases, do not influence their transcription factor function. PMID:21487098

  19. Enzymatic characterization of recombinant ?-amylase in the Drosophila melanogaster species subgroup: is there an effect of specialization on digestive enzyme?

    PubMed

    Commin, Céline; Aumont-Nicaise, Magali; Claisse, Gaëlle; Feller, Georges; Da Lage, Jean-Luc

    2013-01-01

    We performed a comparative study on the enzymological features of purified recombinant ?-amylase of three species belonging to the Drosophila melanogaster species subgroup: D. melanogaster, D. erecta and D. sechellia. D. erecta and D. sechellia are specialist species, with host plant Pandanus candelabrum (Pandanaceae) and Morinda citrifolia (Rubiaceae), respectively. The temperature optima were around 57-60? for the three species. The pH optima were 7.2 for D. melanogaster, 8.2 for D. erecta and 8.5 for D. sechellia. The kcat and Km were also estimated for each species with different substrates. The specialist species D. erecta and D. sechellia display a higher affinity for starch than D. melanogaster. ?-Amylase activity is higher on starch than on glycogen in all species. ?-Amylases of D. erecta and D. sechellia have a higher activity on maltooligosaccharides (G6 and G7) than on starch, contrary to D. melanogaster. Such differences in the enzymological features between the species might reflect adaptation to different ecological niches and feeding habits. PMID:24463528

  20. Inhibitory Potential of Five Traditionally Used Native Antidiabetic Medicinal Plants on ?-Amylase, ?-Glucosidase, Glucose Entrapment, and Amylolysis Kinetics In Vitro

    PubMed Central

    Picot, Carene M. N.; Subratty, A. Hussein; Mahomoodally, M. Fawzi

    2014-01-01

    Five traditionally used antidiabetic native medicinal plants of Mauritius, namely, Stillingia lineata (SL), Faujasiopsis flexuosa (FF), Erythroxylum laurifolium (EL), Elaeodendron orientale (EO), and Antidesma madagascariensis (AM), were studied for possible ?-amylase and ?-glucosidase inhibitory property, glucose entrapment, and amylolysis kinetics in vitro. Only methanolic extracts of EL, EO, and AM (7472.92 ± 5.99, 1745.58 ± 31.66, and 2222.96 ± 13.69??g/mL, resp.) were found to significantly (P < 0.05) inhibit ?-amylase and were comparable to acarbose. EL, EO, AM, and SL extracts (5000??g/mL) were found to significantly (P < 0.05) inhibit ?-glucosidase (between 87.41 ± 3.31 and 96.87 ± 1.37% inhibition). Enzyme kinetic studies showed an uncompetitive and mixed type of inhibition. Extracts showed significant (P < 0.05) glucose entrapment capacities (8 to 29% glucose diffusion retardation index (GDRI)), with SL being more active (29% GDRI) and showing concentration-dependent activity (29, 26, 21, 14, and 5%, resp.). Amylolysis kinetic studies showed that methanolic extracts were more potent inhibitors of ?-amylase compared to aqueous extracts and possessed glucose entrapment properties. Our findings tend to provide justification for the hypoglycaemic action of these medicinal plants which has opened novel avenues for the development of new phytopharmaceuticals geared towards diabetes management. PMID:24723945

  1. Novel insights on the mechanism of action of alpha-amylase inhibitors from the plant defensin family.

    PubMed

    Pelegrini, Patrícia B; Lay, Fung T; Murad, André M; Anderson, Marilyn A; Franco, Octavio L

    2008-11-15

    Plant defensins are small cysteine-rich proteins commonly synthesized in plants, encoded by large multigene families. Most plant defensins that have been characterized to date show potent antifungal and/or bactericidal activities. This report describes VuD1, an unusual defensin that is able to inhibit insect-pest alpha-amylases. VuD1 was cloned from cowpea (Vigna unguiculata) seeds and expressed in a heterologous system. Inhibitory enzyme assays showed that VuD1 efficiently inhibits alpha-amylases from the weevils Acanthoscelides obtectus and Zabrotes subfasciatus, caused low inhibition toward mammalian enzymes and was unable to inhibit the alpha-amylases from Callosobruchus maculatus and Aspergillus fumigatus. To shed some light over the mechanism of action of VuD1, molecular modeling analyses were performed, revealing that the N-terminus of the molecule is responsible for binding with the active site of weevil enzymes. Moreover, models of VuD1 and mammalian enzymes were also generated to elucidate the specificity mechanisms. The data presented herein suggests that this defensin has potential application in the development of transgenic plants for insect pest control. PMID:18498107

  2. Identification of the hydroxyapatite-binding domain of salivary agglutinin.

    PubMed

    Bikker, Floris J; Cukkemane, Nivedita; Nazmi, Kamran; Veerman, Enno C I

    2013-02-01

    The salivary agglutinin glycoprotein (SAG) is present in saliva but is also part of the salivary pellicle, playing a seemingly paradoxical role with regard to bacterial homeostasis. On the one hand, SAG aggregates bacteria in solution, thereby preventing bacterial colonization. On the other hand, when bound to the tooth surface, SAG facilitates bacterial colonization and microbial growth. The protein part of SAG is predominantly composed of conserved scavenger receptor cysteine-rich (SRCR) domains. Previously it was found that bacterial binding and aggregation is mediated via a single peptide loop, designated SRCRP2 (P2), within the SRCR domains of SAG. The current data suggest that the SRCR domains also harbour a hydroxyapatite (HA)-binding moiety, SRCRP3 (P3). The observation that P2 and P3 individually play unique roles in the function of SAGs contributes to our understanding of the dual role of SAGs in bacterial binding. Inspired by the bacterial-modulating capacity of SAGs, we created a P3-polyethylene glycol (PEG) conjugate. It was found that a P3 coating resulted in an increased antifouling activity of 20% compared with the uncoated surface in vitro. An additional PEG moiety resulted in an antifouling activity of up to 40% and 30% for Streptococcus mutans and Staphylococcus epidermidis, respectively. PMID:23331418

  3. Efficient co-displaying and artificial ratio control of ?-amylase and glucoamylase on the yeast cell surface by using combinations of different anchoring domains.

    PubMed

    Inokuma, Kentaro; Yoshida, Takanobu; Ishii, Jun; Hasunuma, Tomohisa; Kondo, Akihiko

    2015-02-01

    Recombinant yeast strains that display heterologous amylolytic enzymes on their cell surface via the glycosylphosphatidylinositol (GPI)-anchoring system are considered as promising biocatalysts for direct ethanol production from starchy materials. For the effective hydrolysis of these materials, the ratio optimization of multienzyme activity displayed on the cell surface is important. In this study, we have presented a ratio control system of multienzymes displayed on the yeast cell surface by using different GPI-anchoring domains. The novel gene cassettes for the cell-surface display of Streptococcus bovis ?-amylase and Rhizopus oryzae glucoamylase were constructed using the Saccharomyces cerevisiae SED1 promoter and two different GPI-anchoring regions derived from Saccharomyces cerevisiae SED1 or SAG1. These gene cassettes were integrated into the Saccharomyces cerevisiae genome in different combinations. Then, the cell-surface ?-amylase and glucoamylase activities and ethanol productivity of these recombinant strains were evaluated. The combinations of the gene cassettes of these enzymes affected the ratio of cell-surface ?-amylase and glucoamylase activities and ethanol productivity of the recombinant strains. The highest ethanol productivity from raw starch was achieved by the strain harboring one ?-amylase gene cassette carrying the SED1-anchoring region and two glucoamylase gene cassettes carrying the SED1-anchoring region (BY-AASS/GASS/GASS). This strain yielded 22.5?±?0.6 g/L of ethanol from 100 g/L of raw starch in 120 h of fermentation. PMID:25432675

  4. Mantle irradiation of the major salivary glands

    SciTech Connect

    Kaplan, P.

    1985-11-01

    Radiation given to the mantle field for treatment of Hodgkin's disease impinges on the submandibular and parotid glands at levels that have been both measured and calculated to be the complete tumor dose. This dosage is above the level of irradiation that has been shown to cause partial or complete loss of salivary gland function.

  5. Microgravity alters the expression of salivary proteins.

    PubMed

    Mednieks, Maija; Khatri, Aditi; Rubenstein, Renee; Burleson, Joseph A; Hand, Arthur R

    2014-06-01

    Spaceflight provides a unique opportunity to study how physiologic responses are influenced by the external environment. Microgravity has been shown to alter the function of a number of tissues and organ systems. Very little, however, is known about how microgravity affects the oral cavity. The rodent model is useful for study in that their salivary gland morphology and physiology is similar to that of humans. Useful also is the fact that saliva, a product of the salivary glands with a major role in maintaining oral health, can be easily collected in humans whereas the glands can be studied in experimental animals. Our working hypothesis is that expression of secretory proteins in saliva will respond to microgravity and will be indicative of the nature of physiologic reactions to travel in space. This study was designed to determine which components of the salivary proteome are altered in mice flown on the US space shuttle missions and to determine if a subset with predictive value can be identified using microscopy and biochemistry methods. The results showed that the expression of secretory proteins associated with beta-adrenergic hormone regulated responses and mediated via the cyclic AMP pathway was significantly altered, whereas that of a number of unrelated proteins was not. The findings are potentially applicable to designing a biochemical test system whereby specific salivary proteins can be biomarkers for stress associated with travel in space and eventually for monitoring responses to conditions on earth. PMID:24984624

  6. Effect of repeated applications of buprofezin and acephate on soil cellulases, amylase, and invertase.

    PubMed

    Raju, M Naga; Venkateswarlu, K

    2014-10-01

    The impact of repeated applications of buprofezin and acephate, at concentrations ranging from 0.25 to 1.0 kg ha(-1), on activities of cellulases, amylase, and invertase in unamended and nitrogen, phosphorous, and potassium (NPK) fertilizer-amended soil planted with cotton was studied. The nontarget effect of selected insecticides, when applied once, twice, or thrice on soil enzyme activities, was dose-dependent; the activities decreased with increasing concentrations of insecticides. However, there was a rapid decline in activities of enzymes after three repeated applications of insecticides in unamended or NPK-amended soil. Our data clearly suggest that insecticides must be applied judiciously in pest management in order to protect the enzymes largely implicated in soil fertility. PMID:24869954

  7. Salivary prions in sheep and deer

    PubMed Central

    Tamgüney, Gültekin; Richt, Jürgen A; Hamir, Amir N; Greenlee, Justin J; Miller, Michael W; Wolfe, Lisa L; Sirochman, Tracey M; Young, Alan J; Glidden, David V; Johnson, Natrina L; Giles, Kurt; DeArmond, Stephen J

    2012-01-01

    Scrapie of sheep and chronic wasting disease (CWD) of cervids are transmissible prion diseases. Milk and placenta have been identified as sources of scrapie prions but do not explain horizontal transmission. In contrast, CWD prions have been reported in saliva, urine and feces, which are thought to be responsible for horizontal transmission. While the titers of CWD prions have been measured in feces, levels in saliva or urine are unknown. Because sheep produce ?17 L/day of saliva and scrapie prions are present in tongue and salivary glands of infected sheep, we asked if scrapie prions are shed in saliva. We inoculated transgenic (Tg) mice expressing ovine prion protein, Tg(OvPrP) mice, with saliva from seven Cheviot sheep with scrapie. Six of seven samples transmitted prions to Tg(OvPrP) mice with titers of ?0.5 to 1.7 log ID50 U/ml. Similarly, inoculation of saliva samples from two mule deer with CWD transmitted prions to Tg(ElkPrP) mice with titers of ?1.1 to ?0.4 log ID50 U/ml. Assuming similar shedding kinetics for salivary prions as those for fecal prions of deer, we estimated the secreted salivary prion dose over a 10-mo period to be as high as 8.4 log ID50 units for sheep and 7.0 log ID50 units for deer. These estimates are similar to 7.9 log ID50 units of fecal CWD prions for deer. Because saliva is mostly swallowed, salivary prions may reinfect tissues of the gastrointestinal tract and contribute to fecal prion shedding. Salivary prions shed into the environment provide an additional mechanism for horizontal prion transmission. PMID:22453179

  8. Modern management of obstructive salivary diseases

    PubMed Central

    Capaccio, P; Torretta, S; Ottaviani, F; Sambataro, G; Pignataro, L

    2007-01-01

    Summary Over the last fifteen years, increasing public demand for minimally-invasive surgery and recent technological advances have led to the development of a number of conservative options for the therapeutic management of obstructive salivary disorders such as calculi and duct stenosis. These include extracorporeal shock-wave lithotripsy, sialoendoscopy, laser intra-corporeal lithotripsy, interventional radiology, the video-assisted conservative surgical removal of parotid and sub-mandibular calculi and botulinum toxin therapy. Each of these techniques may be used as a single therapeutic modality or in combination with one or more of the above-mentioned options, usually in day case or one-day case under local or general anaesthesia. The multi-modal approach is completely successful in about 80% of patients and reduces the need for gland removal in 3%, thus justifying the combination of, albeit, time-consuming and relatively expensive techniques as part of the modern and functional management of salivary calculi. With regard to the management of salivary duct anomalies, such as strictures and kinkings, interventional radiology with fluoroscopically controlled balloon ductoplasty seems to be the most suitable technique despite the use of radiation. Operative sialoendoscopy alone is the best therapeutic option for all mobile intra-luminal causes of obstruction, such as microliths, mucous plugs or foreign bodies, or for the local treatment of inflammatory conditions such as recurrent chronic parotitis or autoimmune salivary disorders. Finally, in the case of failure of one of the above techniques and regardless of the cause of obstruction, botulinum toxin injection into the parenchyma of the salivary glands using colour Doppler ultrasonographic monitoring should be considered before deciding on surgical gland removal. PMID:17957846

  9. Salivary Cortisol Can Replace Free Serum Cortisol Measurements in Patients With Septic Shock

    PubMed Central

    Orlander, Philip R.

    2011-01-01

    Background: There is a renewed interest in adrenal function during severe sepsis. Most studies have used total serum cortisol levels; however, only free serum cortisol is biologically active. The aim of this study was to determine the validity of salivary cortisol levels as a surrogate for free serum cortisol levels during septic shock. Methods: Fifty-seven patients with septic shock were studied to determine the correlation between total serum cortisol and salivary cortisol to free serum cortisol levels. Thirty-eight patients were included in the salivary to free serum cortisol correlation. Salivary cortisol level was tested by enzyme immunoassay. Serum total cortisol, free cortisol, and cortisol-binding globulin (CBG) levels were determined by liquid chromatography-mass spectrometry, equilibrium analysis, and radioimmunoassay, respectively. Results: The mean ± SD age was 56.6 ± 18.5 years. Fifty-seven percent were women. APACHE (Acute Physiology and Chronic Health Evaluation) II score median was 26, Simplified Acute Physiology Score II median was 61, and Sequential Organ Failure Assessment median was 13. The correlation between salivary and free serum cortisol levels was 0.79 (95% CI, 0.63-0.89; P < .0001). The correlation between free serum cortisol and total serum cortisol levels was 0.86 (95% CI, 0.78-0.92; P < .0001). The mean ± SD free serum cortisol level was 2.27 ± 1.64 ?g/dL. The mean ± SD salivary cortisol level was 2.60 ± 2.69 ?g/dL. The mean ± SD total serum cortisol level was 21.56 ± 8.71 ?g/dL. The mean ± SD CBG level was 23.54 ± 8.33 mg/dL. Conclusions: Salivary cortisol level can be used as a surrogate of free serum cortisol level in patients with septic shock with very good correlation. Salivary cortisol testing is noninvasive, easy to perform, and can be conducted daily. Trial registry: ClinicalTrials.gov; No.: NCT00523198; URL: www.clinicaltrials.gov PMID:21816912

  10. In vitro alpha-amylase inhibition and in vivo antioxidant potential of Amaranthus spinosus in alloxan-induced oxidative stress in diabetic rats

    PubMed Central

    Ashok Kumar, B.S.; Lakshman, K.; Nandeesh, R.; Arun Kumar, P.A.; Manoj, B.; Kumar, Vinod; Sheshadri Shekar, D.

    2010-01-01

    Amaranthus spinosus Linn. (Amaranthaceae), commonly known as “Mulluharivesoppu” in Kannada, is used in the Indian traditional system of medicine for the treatment of diabetes. The present study deals with the scientific evaluation of alpha amylase and the antioxidant potential of methanol extract of A. spinosus (MEAS). The aim of this study was to investigate in vitro alpha-amylase enzyme inhibition by CNPG3 (2-chloro-4-nitrophenol ?-d-maltotrioside) and in vivo antioxidant potential of malondialdehyde (MDA), glutathione (GSH), catalase (CAT) and total thiols (TT) in alloxan-induced diabetic rats of a methanolic extract of A. spinosus. Blood sugar was also determined in MEAS-treated alloxan-induced diabetic rats. MEAS showed significant inhibition of alpha-amylase activity and IC50 46.02 ?g/ml. Oral administration of MEAS (200 and 400 mg/kg) for 15 days showed significant reduction in the elevated blood glucose, MDA and restores GSH, CAT and TT levels as compared with a diabetic control. The present study provides evidence that the methanolic extract of A. spinosus has potent alpha amylase, anti-diabetic and antioxidant activities. PMID:23961097

  11. Salivary thyroxine as an estimate of free thyroxine: concise communication

    SciTech Connect

    Elson, M.K.; Morley, J.E.; Shafer, R.B.

    1983-08-01

    To test the hypothesis that the levels of salivary thyroxine (T/sub 4/) reflect those of circulating free T/sub 4/, we developed a radioimmunoassay (RIA) sensitive to low levels of T/sub 4/. Concurrent saliva and serum samples were obtained from 32 euthyroid volunteers, ages 19 to 64. Salivary and serum T/sub 4/ and cortisol levels were measured by RIA. Salivary albumin was measured by nephelometry. Salivary T/sub 4/ levels were higher than predicted. No correlation was found between salivary T/sub 4/ and serum levels of free T/sub 4/ and total T/sub 4/ but there was a significant correlation between salivary T/sub 4/ and albumin (r = 0.82). Salivary cortisol levels agreed with reported results and showed no correlation with salivary albumin. We conclude that salivary levels of drugs and hormones may be strongly affected by protein binding, and caution must be exercised in using salivary levels as an estimate of circulating free levels.

  12. Characterization of maltotriose production by hydrolyzing of soluble starch with ?-amylase from Microbulbifer thermotolerans DAU221.

    PubMed

    Lee, Yong-Suk; Park, Dong-Ju; Choi, Yong-Lark

    2015-05-01

    A maltotriose-producing ?-amylase, AmyA, from a newly isolated bacterial strain Microbulbifer thermotolerans DAU221 was purified and characterized in the heterologous host, Escherichia coli, using the pCold I vector. The amyA gene encoded a 761-residue protein composed of a 33 amino acid secretion signal peptide. The purified ?-amylase with a molecular mass of 80 kDa, approximately, shared a sequence motif characteristic of the glycoside hydrolase family 13. The enzyme was optimally active, at 50 °C in sodium phosphate buffer (pH 6.0), by the traditional one factor-at-a-time method. But the optimal conditions of time, temperature, and pH for production of maltotriose from soluble starch were 1.76 h, 44.95 °C, and pH 6.35 by response surface methodology, respectively. Maltotriose, as the major enzyme reaction product, was analyzed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). The enzyme was found to be inhibited by the addition of 10 mM Cu(2+), Fe(3+), Hg(2+), Zn(2+), and EDTA, but exhibited extreme stability toward hexane. The K m and V max values for the hydrolysis of soluble starch were 1.08 mg/mL and 1.736 mmol maltotriose/mg protein/min, respectively. PMID:25381490

  13. Alpha amylase assisted synthesis of TiO? nanoparticles: structural characterization and application as antibacterial agents.

    PubMed

    Ahmad, Razi; Mohsin, Mohd; Ahmad, Tokeer; Sardar, Meryam

    2015-02-11

    The enzyme alpha amylase was used as the sole reducing and capping agent for the synthesis of TiO2 nanoparticles. The biosynthesized nanoparticles were characterized by X-ray diffraction (XRD) and transmission electron microscopic (TEM) methods. The XRD data confirms the monophasic crystalline nature of the nanoparticles formed. TEM data shows that the morphology of nanoparticles depends upon the enzyme concentration used at the time of synthesis. The presence of alpha amylase on TiO2 nanoparticles was confirmed by FTIR. The nanoparticles were investigated for their antibacterial effect on Staphylococcus aureus and Escherichia coli. The minimum inhibitory concentration value of the TiO2 nanoparticles was found to be 62.50 ?g/ml for both the bacterial strains. The inhibition was further confirmed using disc diffusion assay. It is evident from the zone of inhibition that TiO2 nanoparticles possess potent bactericidal activity. Further, growth curve study shows effect of inhibitory concentration of TiO2 nanoparticles against S. aureus and E. coli. Confocal microscopy and TEM investigation confirm that nanoparticles were disrupting the bacterial cell wall. PMID:25270329

  14. Gibberellin-responsive elements in the promoter of a barley high-pI alpha-amylase gene.

    PubMed Central

    Gubler, F; Jacobsen, J V

    1992-01-01

    Deletion analysis has previously shown that the major gibberellic acid (GA)- and abscisic acid (ABA)-responsive elements in the promoter of a high-pI alpha-amylase gene of barley are located downstream of -174 (Jacobsen and Close, 1991). We have used transient expression assays in barley aleurone protoplasts to identify sequences between -174 and +53 that confer GA and ABA responsiveness on expression of a beta-glucuronidase reporter gene. Using alpha-amylase promoter fragments and synthetic oligonucleotides fused to minimal promoters, we have shown that the hormone-responsive region is located between -174 and -108. A single copy of this region fused to a minimal alpha-amylase promoter (-41) conferred both GA- and ABA-responsive expression on the reporter gene comparable to the positive control, Am(-174)IGN. Multiple copies of this region were able to activate even greater levels of expression. Site-directed mutagenesis was used to determine the functional importance of the conserved motifs (-169pyrimidine box, -143TAACAAA box, and -124TATCCAC box) and nonconserved intervening sequences within the region between -174 and -108. Our results showed that both the TAACAAA and TATCCAC boxes play an important role in GA-regulated expression. We propose that the TAACAAA box is a gibberellin response element, that the TATCCAC box acts cooperatively with the TAACAAA box to give a high level of GA-regulated expression, and that together these motifs form important components of a gibberellin response complex in high-pI alpha-amylase genes. The TAACAAA box also appears to be the site of action of ABA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1477556