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1

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

E-print Network

April 16, 2010 Caffeine and Stress Alter Salivary -Amylase Activity in Young Men *Laura Cousino of Information Sciences and Technology Pennsylvania State University Key words: salivary alpha-amylase, caffeine -amylase assays were conducted by Salimetrics, LLC, State College, PA, where DAG is founder and president

Ritter, Frank

2

Caffeine and stress alter salivary a-amylase activity in young men Laura C. Klein1*, Jeanette M. Bennett1  

E-print Network

Caffeine and stress alter salivary a-amylase activity in young men Laura C. Klein1*, Jeanette M and a psychological stressor on salivary a-amylase (sAA) in healthy young males (age 18­ 30 years) who consumed. Copyright # 2010 John Wiley & Sons, Ltd. key words -- blood pressure; caffeine; heart rate; salivary alpha-amylase

Ritter, Frank

3

Exercise upregulates salivary amylase in humans (Review).  

PubMed

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

Koibuchi, Eri; Suzuki, Yoshio

2014-04-01

4

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

ERIC Educational Resources Information Center

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…

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

2009-01-01

5

The relationship between the level of salivary alpha amylase activity and pain severity in patients with symptomatic irreversible pulpitis  

PubMed Central

Objectives Assessment of dental pain severity is very challenging in dentistry. Previous studies have suggested that elevated salivary alpha amylase may contribute to increased physical stresses. There is a close association between salivary alpha amylase and plasma norepinephrine under stressful physical conditions. The aim of this study was to evaluate the relationship between pain severity and salivary alpha amylase levels in patients with symptomatic irreversible pulpitis. Materials and Methods Thirty-six patients (20 females and 16 males) with severe tooth pain due to symptomatic irreversible pulpitis were selected. The visual analogue scale (VAS) score was used to assess the pain severity in each patient. Unstimulated whole saliva was collected, and the level of alpha amylase activity was assessed by the spectrophotometric method. Statistical analysis was performed using SPSS 13. Results The level of alpha amylase was significantly increased in the saliva in association with pain severity assessed by VAS. The salivary alpha amylase was also elevated with increased age and in males. Conclusions There was a significant correlation between the VAS pain scale and salivary alpha amylase level, which indicates this biomarker may be a good index for the objective assessment of pain intensity. PMID:24010080

Shahriari, Shahriar; Goodarzi, Mohammad Taghi; Moghimbeigi, Abbas; Jazaeri, Mina; Babaei, Parisa

2013-01-01

6

Purification of a novel ?-amylase inhibitor from local Himalayan bean (Phaseolus vulgaris) seeds with activity towards bruchid pests and human salivary amylase.  

PubMed

Six bean (Phaseolus vulgaris L.) cultivars of Himalayan region were analysed for ?- amylase inhibitor activity. The ?-amylase inhibitor from seeds of screened bean cultivar KR-9, showing maximum inhibitory activity was purified using ammonium sulfate precipitation, gel filtration chromatography (Sephadex G-100) and ion exchange chromatography (DEAE-Sephadex). The inhibitor was purified to homogeneity as judged by native-PAGE with 14.22 fold purification and 71.66% recovery. Purified inhibitor consisted of three subunits of molecular weight 15,488, 18,620 and 26,302 daltons, respectively as determined by SDS-PAGE. It was found to be heat stable up to 30 °C-40 °C and had two pH optima of 5.0 and 6.9. Nature of inhibition was found to be of non-competitive type. The purified inhibitor was found to be effective against ?-amylases extracted from larvae of Callosobruchus chinensis, Tribolium castaneum and gut enzyme of Spodoptera littoralis. Larvae of Tribolium castaneum fed on flour mixed with purified inhibitor for 5 days showed 100% larval mortality. Purified ?-amylase inhibitor was also found to inhibit human salivary ?-amylase, suggesting its potential in prevention and therapy of obesity and use as drug design targets for treatment of diabetes. The gene encoding the inhibitor may be used to develop transgenic plants resistant against insect pests. PMID:24966421

Gupta, Mridu; Sharma, Pratima; Nath, Amarjit K

2014-07-01

7

The roles of AMY1 copies and protein expression in human salivary ?-amylase activity.  

PubMed

Salivary ?-amylase (sAA) activity has been extensively investigated in nutrition and psychology. But few studies were performed to assess the role played by sAA gene (AMY1) copies and protein expression in basal and stimulus-induced sAA activity. The sAA activity, amount and AMY1 copy number were determined from 184 saliva samples pre- and post-citric acid stimulation. Our findings showed that citric acid could induce significant increase in sAA activity, total sAA amount, and glycosylated sAA amount, among which the glycosylated sAA amount had the largest response. The correlation analysis showed that AMY1 copy number, total sAA amount and AMY1 copy number×total sAA amount had significantly positive and successively increasing correlations with sAA activity in unstimulated and stimulated saliva, respectively, and furthermore, we observed higher correlations in unstimulated saliva when compared with the corresponding correlations in stimulated saliva. We also observed significant correlations between glycosylated sAA amount and sAA activity in unstimulated and stimulated saliva, respectively. Interestingly, the correlations were higher in stimulated saliva than in unstimulated saliva, and the correlations between glycosylated sAA amount and sAA activity were higher than that of between total sAA amount and sAA activity in stimulated saliva. Moreover, total sAA amount ratio and glycosylated sAA amount ratio showed significantly positive correlation with sAA activity ratio. AMY1 copy number had no correlation with sAA activity ratio. These findings suggested that AMY1 copy number and sAA amount played crucial roles in sAA activity; however, the roles were attenuated after stimulation due to fortified release of glycosylated sAA. PMID:25446200

Yang, Ze-Min; Lin, Jing; Chen, Long-Hui; Zhang, Min; Chen, Wei-Wen; Yang, Xiao-Rong

2015-01-01

8

Psychosocial Stress-Induced Activation of Salivary Alpha-Amylase: An Indicator of Sympathetic Activity?  

Microsoft Academic Search

Assessment of sympathoadrenal medullary system (SAM) activity is only possible to date via measurement of catecholamines in blood plasma or via electrophysiological methods. Both ways of measurement are restricted to endocrinological or psychophysiological laboratories, as both require either immediate freezing of blood samples or complex recording devices. Efforts have therefore been undertaken to find a method comparable to salivary cor-

NICOLAS ROHLEDER; URS M. NATER; JUTTA M. WOLF; ULRIKE EHLERT; CLEMENS KIRSCHBAUM

2004-01-01

9

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

PubMed

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

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

2014-08-01

10

Monitoring salivary amylase activity is useful for providing timely analgesia under sedation  

PubMed Central

AIM: To detect the criteria and cause of elevated salivary amylase activity (sAMY) in patients undergoing endoscopic submucosal dissection (ESD) under sedation. METHODS: A total of 41 patients with early gastric cancer removed via ESD under deep sedation (DS) were enrolled. The perioperative sAMY, which was shown as sympathetic excitements (SE), was measured. The time at which a patient exhibited a relatively increased rate of sAMY compared with the preoperative baseline level (IR, %) ? 100% (twice the actual value) was assumed as the moment when the patient received SE. Among the 41 patients, we focused on 14 patients who exhibited an IR ? 100% at any time that was associated with sAMY elevation during ESD (H-group) and examined whether any particular endoscopic procedures can cause SE by simultaneously monitoring the sAMY level. If a patient demonstrated an elevated sAMY level above twice the baseline level, the endoscopic procedure was immediately stopped. In the impossible case of discontinuance, analgesic medicines were administered. This study was performed prospectively. RESULTS: A total of 26 episodes of sAMY eruption were considered moments of SE in the H-group. The baseline level of sAMY significantly increased in association with an IR of > 100% at 5 min, with a significant difference (IR immediately before elevation/IR at elevation of sAMY = 8.72 ± 173/958 ± 1391%, P < 0.001). However, effective intervention decreased the elevated sAMY level immediately within only 5 min, with a significant difference (IR at sAMY elevation/immediately after intervention = 958 ± 1391/476 ± 1031, P < 0.001). The bispectral indices, systolic blood pressure and pulse rates, which were measured at the same time, remained stable throughout the ESD. Forceful endoscopic insertion or over insufflation was performed during 22 of the 26 episodes. Release of the gastric wall tension and/or the administration of analgesic medication resulted in the immediate recovery of the elevated sAMY level, independent of body movement. CONCLUSION: By detecting twice the actual sAMY based on the preoperative level, the release of the gastric wall tension or the administration of analgesic agents should be considered. PMID:24932376

Uesato, Masaya; Nabeya, Yoshihiro; Akai, Takashi; Inoue, Masahito; Watanabe, Yoshiyuki; Horibe, Daisuke; Kawahira, Hiroshi; Hayashi, Hideki; Matsubara, Hisahiro

2014-01-01

11

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

PubMed Central

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

Mandel, Abigail L.

2012-01-01

12

Klein, L. C., Whetzel, C. A., Bennett, J. M., Ritter, F. E., & Granger, D. A. (2006 March). Effects of caffeine and stress on salivary alpha-amylase in young men: A salivary biomarker  

E-print Network

of caffeine and stress on salivary alpha-amylase in young men: A salivary biomarker of sympathetic activity ON SALIVARY ALPHA-AMYLASE IN YOUNG MEN: A SALIVARY BIOMARKER OF SYMPATHETIC ACTIVITY Laura Cousino Klein, Courtney A. Whetzel, Jeanette M. Bennett, Frank E. Ritter, Douglas A. Granger, Penn State University Alpha-amylase

Ritter, Frank

13

Characterization of salivary alpha-amylase binding to Streptococcus sanguis  

SciTech Connect

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.

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

1989-09-01

14

Partial characterization of ?-amylase in the salivary glands of Lygus hesperus and L. lineolaris  

Microsoft Academic Search

The ?-amylases in the salivary glands of Lygus hesperus Knight and L. lineolaris (Palisot de Beauvois) were isolated and purified by ion exchange chromatography, and by isoelectric focusing, respectively. The ?-amylase from L. hesperus had an isoelectric point (pI) of 6.25, and a pH optimum of 6.5. The specific activity of ?-amylases in the salivary glands of L. hesperus was

Fanrong Zeng; Allen C Cohen

2000-01-01

15

Salivary amylase and stress during stressful environment: three Mars analog mission crews study.  

PubMed

After the establishment of the space age physicians, human factors engineers, neurologist and psychologists and their special attention to work on people's capability to meet up the physical, psychological, neuroscience and interpersonal strains of working in space, it has been regarded as an issue that seeks urgent consideration. Not study was conducted on effect of simulated Mars analog environment on stress and salivary amylase. So, this study aimed to confirm whether salivary amylase is act as stress biomarker in crew members who took part in Mars analog mission in an isolated and stressful environment. The 18 crew members were selected who took part in Mars Analog Research Station, Utah. Salivary amylase was measured using a biosensor of salivary amylase monitor and State-Trait Anxiety Inventory score at pre-extravehicular activity, post-extravehicular activity and on before mission. The state and trait anxiety scores at pre-extravehicular activity for each commander were elevated as compared to after extravehicular activity. There were significant differences in the state and trait anxiety scores between before extravehicular activity and after extravehicular activity of Commander and other members, also there were significant differences in values of before-extravehicular activity between commanders and other members. There were significant differences in values of salivary amylase at before extravehicular activity and after extravehicular activity between commander group and other members. There was significant correlation between salivary amylase and state and trait anxiety scores in all groups. Measuring salivary amylase level could be useful for stress assessment of crew members and population working in a stressful and isolated environment. PMID:22554904

Rai, Balwant; Kaur, Jasdeep; Foing, Bernard H

2012-06-14

16

Salivary gland enlargement and elevated serum amylase in bulimia nervosa  

Microsoft Academic Search

Background: Clinical reports have described salivary gland enlargement in bulimia nervosa, particularly in patients with elevated serum amylase concentration. The goal of the current study was to provide a controlled comparison of salivary gland size in patients with bulimia nervosa and healthy volunteers.Methods: Subjects included 17 women with bulimia nervosa and 21 healthy female control subjects. Dimensions of the parotid

Eran D Metzger; Jeffrey M Levine; Colin R McArdle; Barbara E Wolfe; David C Jimerson

1999-01-01

17

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

PubMed Central

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

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

2014-01-01

18

Words number: 25491 Assessment of salivary alpha-amylase -a stress biomarker -in2  

E-print Network

1 Words number: 25491 Assessment of salivary alpha-amylase - a stress biomarker - in2 pregnant patients.3 4 Running tittle: Salivary alpha-amylase: a stress biomarker in pregnant patients5 6 Jean, Psychological5 Saliva6 Salivary alpha-Amylases7 Caesarean Section8 9 10 inserm-00847842,version1-24Jul2013 #12

Boyer, Edmond

19

Salivary alpha amylase and cortisol responses to different stress tasks: Impact of sex  

Microsoft Academic Search

Neuro-endocrine markers such as salivary alpha amylase (sAA) and cortisol (CORT) play an important role in establishing human responses to stressful events. Whereas sAA levels reflect sympathetic system activity, salivary cortisol appears to be a valid measure for HPA axis activity. Although many studies looked at either sAA or CORT responses in reaction to stress, work still has to be

Anda H. van Stegeren; Oliver T. Wolf; Merel Kindt

2008-01-01

20

Salivary Amylase Promotes Adhesion of Oral Streptococci to Hydroxyapatite  

Microsoft Academic Search

Recent studies have demonstrated that several species of oral streptococci, such as Streptococcus gordonii, bind soluble salivary a-amylase. The goal of the present study was to determine if amylase immobilized onto a surface such as hydroxyapatite can serve as an adhesion receptor for S. gordonii. Initially, human parotid saliva was fractionated on Bio-Gel P60, and fractions were screened for their

F. A. Scannapieco; G. I. Torres; M. J. Levine

1995-01-01

21

Liquid chromatographic assay of the relative activities of serum pancreatic and salivary alpha-amylase using reductively pyridylaminated maltopentaose as a fluorescent substrate.  

PubMed

A method for the high-performance liquid chromatographic assay of the relative activities of serum pancreatic and salivary alpha-amylase has been developed, using maltopentaose reductively aminated with 2-aminopyridine as a fluorescent substrate. Both enzymes showed similar modes of action, cleaving the second and the third (from the non-reducing terminal) interglycosidic linkages of this substrate. However, the relative ease of cleavage of these two sites by the pancreatic enzyme was significantly different from that by the salivary enzyme. Therefore, determination of the molar ratio of the cleavage products by HPLC could lead to estimation of the activity ratio of these enzymes. The optimum chromatographic conditions for HPLC were as follows: column, LiChrosorb RP-18 (Merck, 7 microns, 250 X 4 mm I.D.); column temperature, ambient; eluent, 0.01% orthophosphoric acid-acetonitrile (4:1, v/v) containing 2.4 mM sodium laurylsulphate; flow-rate, 1.0 ml/min; wavelengths for fluorimetric detection, 320 nm (excitation)/400 nm (emission). The problem of interference by serum alpha-glucosidase was solved by specific inhibition with tris (hydroxymethyl) aminomethane and erythritol. The data obtained by the proposed method correlated well with those produced by the conventional method based on electrophoresis. PMID:3499444

Honda, S; Ohmura, T; Segawa, Y; Minematsu, T; Kakehi, K

1987-08-01

22

The Effect of a Brief Salivary ?-Amylase Exposure During Chewing on Subsequent in Vitro Starch Digestion Curve Profiles  

PubMed Central

There is inconsistency between current in vitro digestion methods with regard to accommodation of a (salivary) ?-amylase exposure during the oral phase. The effect of a salivary ?-amylase pre-exposure on subsequent in vitro starch digestion curve profiles for various foods was investigated. Foods were chewed, expectorated and the boluses left to rest for 0–15 min. During pancreatic digestion, aliquots were taken and hydrolysis curves constructed for comparison against those of the same foods comminuted with a manually-operated chopper, hence spared exposure to saliva. Hydrolysate aliquots taken at T0 (time zero) of the digestion of chewed samples contained higher levels of glucose and dextrins compared with chopped samples. Pancreatin activity immediately overwhelmed differences in sugar released due to salivary amylase activity. Within 10 min no differences were detectable between hydrolysis curves for chewed and chopped foods. Salivary amylase pretreatment does not contribute to the robustness or relative accuracy of in vitro methods. PMID:21152272

Woolnough, James W.; Bird, Anthony R.; Monro, John A.; Brennan, Charles S.

2010-01-01

23

Human salivary alpha-amylase reactivity in a psychosocial stress paradigm  

Microsoft Academic Search

Biological indicators for stress reactions are valuable markers in psychophysiological research and clinical practice. Since the release of salivary enzyme alpha-amylase was reported to react to physiological and psychological stressors, we set out to investigate human salivary alpha-amylase changes employing a reliable laboratory stress protocol to investigate the reactivity of salivary alpha-amylase to a brief period of psychosocial stress.In a

Urs M. Nater; Nicolas Rohleder; Jens Gaab; Simona Berger; Andreas Jud; Clemens Kirschbaum; Ulrike Ehlert

2005-01-01

24

Salivary ?-Amylase Reactivity to Infant Crying in Maltreating Mothers.  

PubMed

Deviant physiological reactivity to infant stimuli has been suggested to underlie maladaptive parenting behavior. Our study involved 44 maltreating and 42 non-maltreating mothers. During a standardized cry paradigm, mothers listened to nine cry sounds of varying pitches. Saliva was collected at baseline, after each cry sound, and after a recovery episode. Salivary ?-amylase (sAA) as a marker of autonomic nervous system (ANS) activity was assayed from saliva samples. Maltreating mothers showed lower overall sAA levels and an attenuated reactivity pattern to infant crying as compared to non-maltreating mothers. No effect of type of maltreatment (neglect only vs. neglect and abuse) was found. Furthermore, positive correlations between sAA and heart rate (HR) for non-maltreating mothers differed significantly from non-significant correlations between sAA and HR for maltreating mothers. This suggests anomalous asynchrony between different aspects of the ANS in maltreating mothers. Results indicate a lack of functional autonomic (re)activity as a contributing risk factor to child maltreatment. PMID:25257947

Reijman, Sophie; Alink, Lenneke R A; Compier-de Block, Laura H C G; Werner, Claudia D; Maras, Athanasios; Rijnberk, Corine; van IJzendoorn, Marinus H; Bakermans-Kranenburg, Marian J

2014-09-26

25

Interparental Aggression and Parent-Adolescent Salivary Alpha Amylase Symmetry  

PubMed Central

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

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

2010-01-01

26

Chronic stress, salivary cortisol, and ?-amylase in children with asthma and healthy children  

Microsoft Academic Search

The present study examined whether chronic stress is related to daily life levels of salivary ?-amylase (sAA), a marker for sympathetic activity, and cortisol in healthy children versus children with asthma.Children's sAA and cortisol levels were measured repeatedly over 2 days. Chronic stress measures included interviews with children about chronic home life stress and interviews with parents about one marker

Jutta M. Wolf; Erin Nicholls; Edith Chen

2008-01-01

27

Detection of pulmonary amylase activity in exhaled breath condensate.  

PubMed

Amylase activity in exhaled breath condensate (EBC) is usually interpreted as an indication of oropharyngeal contamination despite the fact that amylase can be found in pulmonary excretions. The aim of this study was to recruit and refine an amylase assay in order to detect amylase activity in any EBC sample and to develop a method to identify EBC samples containing amylase of pulmonary origin. EBC was collected from 40 volunteers with an EcoScreen condenser. Amylase assays and methods to discriminate between oropharyngeal and pulmonary proteins were tested and developed using matched EBC and saliva samples. Our refined 2-chloro-4-nitrophenyl-?-D-maltotriosid (CNP-G3) assay was 40-fold more sensitive than the most sensitive commercial assay and allowed detection of amylase activity in 30 µl of EBC. We developed a dot-blot assay which allowed detection of salivary protein in saliva diluted up to 150 000-fold. By plotting amylase activity against staining intensity we identified a few EBC samples with high amylase activity which were aligned with diluted saliva. We believe that EBC samples aligned with diluted saliva contain amylase activity introduced during EBC collection and that all other EBC samples contain amylase activity of pulmonary origin and are basically free of oropharyngeal protein contamination. PMID:24287549

Zweifel, M; Rechsteiner, T; Hofer, M; Boehler, A

2013-12-01

28

Metabolism of glycosylated human salivary amylase: in vivo plasma clearance by rat hepatic endothelial cells and in vitro receptor mediated pinocytosis by rat macrophages  

SciTech Connect

Salivary-type amylase normally comprises about 60% of the amylase activity in human serum, but only a small fraction is a glycosylated isoenzyme (amylase A). In contrast, 1/3 of amylase in human saliva is glycosylated. Since glycosylation can affect circulatory clearance, we studied the clearance of amylase A in rats and its uptake by rat alveolar macrophages. Following intravenous injection, /sup 125/I-labeled amylase A disappeared rapidly from plasma (t 1/2 . 9 min) and accumulated in the liver. Simultaneous injection of mannose-albumin slowed its clearance to a rate comparable to that of /sup 125/I-labeled nonglycosylated salivary amylase (t 1/2 . 45 min). In contrast, galactose-albumin had no effect. Electron microscope autoradiography of the liver following injection of /sup 125/I-labeled amylase A revealed a localization of grains over the hepatic endothelial cells. In vitro studies indicated that amylase A is taken up by alveolar macrophages via receptor-mediated pinocytosis. Uptake was linear over time, saturable, and inhibited by mannan and mannose-albumin, but not by galactose-albumin. We conclude that amylase A, which is a naturally occurring human glycoprotein with at most three terminal L-fucose residues per molecule, is recognized in rats by a mannose receptor located on hepatic endothelial cells. We speculate that this receptor, by rapidly clearing circulating amylase A, may be responsible for the low level of amylase A in human serum.

Niesen, T.E.; Alpers, D.H.; Stahl, P.D.; Rosenblum, J.L.

1984-09-01

29

Salivary alpha-amylase and cortisol responsiveness following electrically stimulated physical stress in bipolar disorder patients  

PubMed Central

Background Bipolar disorder (BP) is often associated with a change in hypothalamus– pituitary–adrenal axis function change due to chronic stress. Salivary ?-amylase (sAA) levels increase in response to psychosocial stress and thus function as a marker of sympathoadrenal medullary system activity. However, sAA has been studied less often than salivary cortisol in BP patients. Method We measured Profile of Mood States and State-Trait Anxiety Inventory scores, heart rate variability, and salivary cortisol levels during electrical stimulation stress in 25 BP patients and 22 healthy volunteers. Results Tension–anxiety, depression–dejection, anger–hostility, fatigue, and confusion scores in BP patients significantly increased compared with those of the healthy controls. In contrast, the vigor scores of BP patients significantly decreased compared with those of the healthy controls. Significant difference in the sAA levels was observed between BP patients and healthy controls. sAA of female patients was significantly higher than that of female healthy controls, and sAA in male patients tended to be higher than that of male healthy controls. No difference in salivary cortisol was observed between BP patients and the healthy controls. Only three time points were measured before and after the electrical stimulation stress. Furthermore, sAA secretion by BP patients increased before and after electrical stimulation. Conclusion These preliminary results suggest that sAA may be a useful biological marker for BP patients. PMID:24353422

Tanaka, Yoshihiro; Maruyama, Yoshihiro; Ishitobi, Yoshinobu; Kawano, Aimi; Ando, Tomoko; Ikeda, Rie; Inoue, Ayako; Imanaga, Junko; Okamoto, Shizuko; Kanehisa, Masayuki; Ninomiya, Taiga; Tsuru, Jusen; Akiyoshi, Jotaro

2013-01-01

30

Feeding activity, salivary amylase activity, and superficial damage to soybean seed by adult Edessa meditabunda (F.) and Euschistus heros (F.) (Hemiptera: Pentatomidae).  

PubMed

Greenhouse and laboratory studies were conducted to evaluate feeding activity and superficial damage to soybean seed by the brown-winged stink bug, Edessa meditabunda (F.), and the Neotropical brown stink bug, Euschistus heros (F.). Soybean plants (cv. BRS 282), at R6 stage of development were used. Thirty pairs of each species were used individually for 48 h. Two daily observations (9:00 AM and 3:00 PM) were taken to record the number of bugs (feeding/resting) on plant parts. Harvested seeds imbibed in tetrazolium solution were photographed for measurement of the damaged surface. Adult E. meditabunda significantly preferred soybean stems (19.7 bugs) to pods (2.7). Feeding/resting was similar at 9:00 AM (mean number of 28.0 bugs) and 3:00 PM (24.3). Euschistus heros equally fed/stayed on stems (7.3 bugs) and pods (6.9), although most bugs (12.3) remained on the cage net; feeding/resting on all plant structures amounted to 13.7 bugs at 9:00 AM and 17.7 bugs at 3:00 PM. Amylase activity was greater for E. heros (41.61 ± 0.89 U/mg) and almost none for E. meditabunda (2.35 ± 0.14 U/mg). The superficial damage to seeds was significantly greater for E. meditabunda (22. 9 mm(2)) compared to E. heros (12.5 mm(2)). However, E. meditabunda caused less shrinkage of the seed tegument, while E. heros damage was deeper and seeds showed reduction in size. PMID:23950088

Silva, F A C; da Silva, J J; Depieri, R A; Panizzi, Antônio Ricardo

2012-10-01

31

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

ERIC Educational Resources Information Center

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…

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

2012-01-01

32

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

ERIC Educational Resources Information Center

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…

Marini, Isabella

2005-01-01

33

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

ERIC Educational Resources Information Center

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…

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

2010-01-01

34

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

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…

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

35

Salivary nitric oxide and alpha-amylase as indexes of training intensity and load.  

PubMed

This study examined the variation in salivary nitric oxide (NO), alpha-amylase (sAA) and serum markers of muscle injury during 21 weeks of training in elite swimmers. Samples of saliva and blood were collected once a month during 5 months from 11 male professional athletes during their regular training season. The variation in each marker throughout the 21 weeks was compared with the dynamics of training volume, intensity and load. Unstimulated whole saliva was assessed for NO and sAA whereas venous blood was assessed for lactate dehydrogenase, creatine kinase, and ?-glutamyltransferase. Nitric oxide and sAA showed a proportional response to the intensity of training. However, whereas the concentration of NO increased across the 21 weeks, the activity of sAA decreased. Similar variations in the concentration of NO and the markers of muscle injury were also observed. The higher concentration of NO might be attributed to changes in haemodynamics and muscle regenerative processes. On the other hand, autonomic regulation towards parasympathetic predominance might have been responsible for the decrease in sAA activity. These findings provide appealing evidence for the utilization of salivary constituents in sports medicine to monitor training programmes. PMID:22960992

Diaz, M M; Bocanegra, O L; Teixeira, R R; Soares, S S; Espindola, F S

2013-01-01

36

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

PubMed Central

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

Bhandary, Rahul; Thomas, Biju; Kumari, Suchetha

2014-01-01

37

Salivary alpha-amylase and cortisol responsiveness following electrical stimulation stress in obsessive-compulsive disorder patients.  

PubMed

Salivary ?-amylase (sAA) serves as a marker of sympathoadrenal medullary system (SAM) activity. Salivary AA has not been extensively studied in obsessive-compulsive disorder (OCD) patients. In the current study, 45 OCD patients and 75 healthy volunteers were assessed with the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS), the Profile of Mood State (POMS), and the State-Trait Anxiety Inventory (STAI). Measures of heart rate variability (HRV), sAA, and salivary cortisol were also obtained following the application of electrical stimulation stress. The Y-BOCS and POMS Tension-Anxiety, Depression-Dejection, Anger-Hostility, Fatigue, and Confusion scores were significantly increased in patients with OCD compared with healthy controls. In contrast, Vigor scores were significantly decreased in patients with OCD relative to scores in healthy controls. There was no difference in HRV between the patients and the controls. Salivary AA levels in female and male OCD patients were significantly elevated relative to controls both before and after electrical stimulation. In contrast, there were no differences in salivary cortisol levels between OCD patients and controls. The elevated secretion of sAA before and after stimulation may suggest an increased responsiveness to novel and uncontrollable situations in patients with OCD. An increase in sAA might be a characteristic change of OCD. PMID:23266021

Kawano, Aimi; Tanaka, Yoshihiro; Ishitobi, Yoshinobu; Maruyama, Yoshihiro; Ando, Tomoko; Inoue, Ayako; Okamoto, Shizuko; Imanaga, Junko; Kanehisa, Masayuki; Higuma, Haruka; Ninomiya, Taiga; Tsuru, Jusen; Akiyoshi, Jotaro

2013-08-30

38

Measurements of Salivary Alpha Amylase and Salivary Cortisol in Hominoid Primates Reveal Within-Species Consistency and Between-Species Differences  

PubMed Central

Salivary alpha amylase (sAA) is the most abundant enzyme in saliva. Studies in humans found variation in enzymatic activity of sAA across populations that could be linked to the copy number of loci for salivary amylase (AMY1), which was seen as an adaptive response to the intake of dietary starch. In addition to diet dependent variation, differences in sAA activity have been related to social stress. In a previous study, we found evidence for stress-induced variation in sAA activity in the bonobos, a hominoid primate that is closely related to humans. In this study, we explored patterns of variation in sAA activity in bonobos and three other hominoid primates, chimpanzee, gorilla, and orangutan to (a) examine if within-species differences in sAA activity found in bonobos are characteristic for hominoids and (b) assess the extent of variation in sAA activity between different species. The results revealed species-differences in sAA activity with gorillas and orangutans having higher basal sAA activity when compared to Pan. To assess the impact of stress, sAA values were related to cortisol levels measured in the same saliva samples. Gorillas and orangutans had low salivary cortisol concentrations and the highest cortisol concentration was found in samples from male bonobos, the group that also showed the highest sAA activity. Considering published information, the differences in sAA activity correspond with differences in AMY1 copy numbers and match with general features of natural diet. Studies on sAA activity have the potential to complement molecular studies and may contribute to research on feeding ecology and nutrition. PMID:23613746

Behringer, Verena; Borchers, Claudia; Deschner, Tobias; Möstl, Erich; Selzer, Dieter; Hohmann, Gottfried

2013-01-01

39

Expression of the human amylase genes: Recent origin of a salivary amylase promoter from an actin pseudogene  

SciTech Connect

The human genes encoding salivary amylase (AMY1) and pancreatic amylase (AMY2) are nearly identical in structure and sequence. The authors have used ribonuclease protection studies to identify the functional gene copies in this multigene family. Riboprobes derived from each gene were hybridized to RNA from human pancreas, parotid and liver. The sizes of the protected fragments demonstrated that both pancreatic genes are expressed in pancreas. One of the pancreatic genes, AMY2B, is also transcribed at a low level in liver, but not from the promoter used in pancreas. AMY1 transcripts were detected in parotid, but not in pancreas or liver. Unexpected fragments protected by liver RNA led to the discovery that the 5{prime} regions of the five human amylase genes contain a processed {gamma}-actin pseudogene. The promoter and start site for transcription of AMY1 are recently derived from the 3{prime} untranslated region of {gamma}-actin. In addition, insertion of an endogenous retrovirus has interrupted the {gamma}-actin pseudogene in four of the five amylase genes.

Samuelson, L.C.; Gumucio, D.L.; Meisler, M.H. (Univ. of Michigan, Ann Arbor (USA)); Wiebauer, K. (Friedrich Miescher Institut, Basel (Switzerland))

1988-09-12

40

Differences in Salivary Alpha-Amylase and Cortisol Responsiveness following Exposure to Electrical Stimulation versus the Trier Social Stress Tests  

PubMed Central

Background Cortisol is an essential hormone in the regulation of the stress response along the HPA axis, and salivary cortisol has been used as a measure of free circulating cortisol levels. Recently, salivary alpha-amylase (sAA) has also emerged as a novel biomarker for psychosocial stress responsiveness within the sympathetic adrenomedullary (SAM) system. Principal Findings We measured sAA and salivary cortisol in healthy volunteers after exposure to the Trier Social Stress Test (TSST) and electric stimulation stress. One hundred forty-nine healthy volunteers participated in this study. All subjects were exposed to both the TSST and electric stimulation stress on separate days. We measured sAA and salivary cortisol levels three times immediately before, immediately after, and 20 min after the stress challenge. The State (STAI-S) and Trait (STAI-T) versions of the Spielberger Anxiety Inventory test and the Profile of Mood State (POMS) tests were administered to participants before the electrical stimulation and TSST protocols. We also measured HF, LF and LF/HF Heart Rate Variability ratio immediately after electrical stimulation and TSST exposure. Following TSST exposure or electrical stimulation, sAA levels displayed a rapid increase and recovery, returning to baseline levels 20 min after the stress challenge. Salivary cortisol responses showed a delayed increase, which remained significantly elevated from baseline levels 20 min after the stress challenge. Analyses revealed no differences between men and women with regard to their sAA response to the challenges (TSST or electric stimulations), while we found significantly higher salivary cortisol responses to the TSST in females. We also found that younger subjects tended to display higher sAA activity. Salivary cortisol levels were significantly correlated with the strength of the applied electrical stimulation. Conclusions These preliminary results suggest that the HPA axis (but not the SAM system) may show differential response patterns to distinct kinds of stressors. PMID:22859941

Maruyama, Yoshihiro; Kawano, Aimi; Okamoto, Shizuko; Ando, Tomoko; Ishitobi, Yoshinobu; Tanaka, Yoshihiro; Inoue, Ayako; Imanaga, Junko; Kanehisa, Masayuki; Higuma, Haruka; Ninomiya, Taiga; Tsuru, Jusen; Hanada, Hiroaki; Akiyoshi, Jotaro

2012-01-01

41

Zeamatin Inhibits Trypsin and ?-Amylase Activities  

PubMed Central

Zeamatin is a 22-kDa protein isolated from Zea mays that has antifungal activity against human and plant pathogens. Unlike other pathogenesis-related group 5 proteins, zeamatin inhibits insect ?-amylase and mammalian trypsin activities. It is of clinical significance that zeamatin did not inhibit human ?-amylase activity and inhibited mammalian trypsin activity only at high molar concentrations. PMID:11319124

Schimoler-O'Rourke, Rebecca; Richardson, Michael; Selitrennikoff, Claude P.

2001-01-01

42

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

PubMed

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

Koh, David; Ng, Vivian; Naing, Lin

2014-01-01

43

Associations between salivary alpha-amylase and catecholamines - A multilevel modeling approach.  

PubMed

Salivary alpha-amylase (sAA) serves as indicator for sympathetic activity. However, previous findings on the association between aggregated sAA and other sympathetic markers, namely norepinephrine and epinephrine, were mixed. We therefore assumed that time-sensitive statistical analyses might help identifying possible associations of sAA and catecholamines. Data from two studies were analyzed. In Study 1, 13 men were examined in a randomized repeated within-subjects double-blind study with yohimbine/placebo. In Study 2, 30 men were randomized in a repeated within-subjects design to psychosocial stress/rest. Associations of repeatedly assessed sAA, norepinephrine, and epinephrine in blood were analyzed using multilevel modeling. Over the time course, sAA was significantly associated with the catecholamines (Study 1: R(2)=.43, Study 2: R(2)=.09) and both served as mediators of sAA increases. Additional exploratory analyses suggest stronger associations during challenge/stress than during placebo/rest. These findings further support sAA as marker of sympathetic activity. PMID:25132576

Ditzen, Beate; Ehlert, Ulrike; Nater, Urs M

2014-12-01

44

Altered salivary alpha-amylase awakening response in Bosnian War refugees with posttraumatic stress disorder.  

PubMed

In posttraumatic stress disorder (PTSD), chronic activation of the sympathetic nervous system (SNS) has been suggested. No study so far has investigated diurnal secretion patterns of salivary alpha-amylase (sAA) in PTSD, a promising candidate for non-invasive assessment of SNS activity. We compared sAA diurnal profiles between a group of Bosnian War refugees with PTSD and a healthy control group, and further analyzed for associations with psychiatric symptoms and glucocorticoid (GC) sensitivity of inflammatory regulation. PTSD patients showed a sAA awakening response profile that was opposite to those seen in healthy controls, i.e. an increase instead of a sharp decrease. Patterns of sAA secretion were further positively associated with psychiatric symptoms of PTSD. Finally, higher sAA awakening responses were associated with higher GC sensitivity of inflammatory cytokine production. These findings are in line with altered SNS function in PTSD, and lend further support for employing assessment of diurnal sAA profiles as non-invasive biomarkers in stress-related disease. PMID:22001009

Thoma, Myriam Verena; Joksimovic, Ljiljana; Kirschbaum, Clemens; Wolf, Jutta Manuela; Rohleder, Nicolas

2012-06-01

45

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

PubMed Central

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

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

2012-01-01

46

Diurnal profiles of salivary cortisol and alpha-amylase change across the adult lifespan: Evidence from repeated daily life assessments  

PubMed Central

Summary Salivary cortisol and alpha-amylase are known to have distinctive diurnal profiles. However, little is known about systematic changes in these biomarkers across the adult lifespan. In a study of 185 participants (aged 20–81 years), time-stamped salivary cortisol and alpha-amylase were collected 7 times/day over 10 days. Samples were taken upon waking, 30 minutes later, and then approximately every 3 hours until 9pm. Multilevel models showed that older age was associated with increased daily cortisol secretion as indicated by greater area under the curve, attenuated wake-evening slopes, and more pronounced cortisol awakening responses. Further, older age was related to greater daily alpha-amylase output and attenuated wake-evening slopes. No age differences were observed regarding the alpha-amylase awakening response. Our findings may contribute to a better understanding of age-related differences in functioning of stress-related systems. PMID:24099860

Nater, Urs M.; Hoppmann, Christiane A.; Scott, Stacey B.

2013-01-01

47

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

ERIC Educational Resources Information Center

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

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

2010-01-01

48

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

PubMed Central

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

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

2013-01-01

49

Low copy number of the salivary amylase gene predisposes to obesity.  

PubMed

Common multi-allelic copy number variants (CNVs) appear enriched for phenotypic associations compared to their biallelic counterparts. Here we investigated the influence of gene dosage effects on adiposity through a CNV association study of gene expression levels in adipose tissue. We identified significant association of a multi-allelic CNV encompassing the salivary amylase gene (AMY1) with body mass index (BMI) and obesity, and we replicated this finding in 6,200 subjects. Increased AMY1 copy number was positively associated with both amylase gene expression (P = 2.31 × 10(-14)) and serum enzyme levels (P < 2.20 × 10(-16)), whereas reduced AMY1 copy number was associated with increased BMI (change in BMI per estimated copy = -0.15 (0.02) kg/m(2); P = 6.93 × 10(-10)) and obesity risk (odds ratio (OR) per estimated copy = 1.19, 95% confidence interval (CI) = 1.13-1.26; P = 1.46 × 10(-10)). The OR value of 1.19 per copy of AMY1 translates into about an eightfold difference in risk of obesity between subjects in the top (copy number > 9) and bottom (copy number < 4) 10% of the copy number distribution. Our study provides a first genetic link between carbohydrate metabolism and BMI and demonstrates the power of integrated genomic approaches beyond genome-wide association studies. PMID:24686848

Falchi, Mario; El-Sayed Moustafa, Julia Sarah; Takousis, Petros; Pesce, Francesco; Bonnefond, Amélie; Andersson-Assarsson, Johanna C; Sudmant, Peter H; Dorajoo, Rajkumar; Al-Shafai, Mashael Nedham; Bottolo, Leonardo; Ozdemir, Erdal; So, Hon-Cheong; Davies, Robert W; Patrice, Alexandre; Dent, Robert; Mangino, Massimo; Hysi, Pirro G; Dechaume, Aurélie; Huyvaert, Marlène; Skinner, Jane; Pigeyre, Marie; Caiazzo, Robert; Raverdy, Violeta; Vaillant, Emmanuel; Field, Sarah; Balkau, Beverley; Marre, Michel; Visvikis-Siest, Sophie; Weill, Jacques; Poulain-Godefroy, Odile; Jacobson, Peter; Sjostrom, Lars; Hammond, Christopher J; Deloukas, Panos; Sham, Pak Chung; McPherson, Ruth; Lee, Jeannette; Tai, E Shyong; Sladek, Robert; Carlsson, Lena M S; Walley, Andrew; Eichler, Evan E; Pattou, Francois; Spector, Timothy D; Froguel, Philippe

2014-05-01

50

Amylase activity is associated with AMY2B copy numbers in dog: implications for dog domestication, diet and diabetes.  

PubMed

High amylase activity in dogs is associated with a drastic increase in copy numbers of the gene coding for pancreatic amylase, AMY2B, that likely allowed dogs to thrive on a relatively starch-rich diet during early dog domestication. Although most dogs thus probably digest starch more efficiently than do wolves, AMY2B copy numbers vary widely within the dog population, and it is not clear how this variation affects the individual ability to handle starch nor how it affects dog health. In humans, copy numbers of the gene coding for salivary amylase, AMY1, correlate with both salivary amylase levels and enzyme activity, and high amylase activity is related to improved glycemic homeostasis and lower frequencies of metabolic syndrome. Here, we investigate the relationship between AMY2B copy numbers and serum amylase activity in dogs and show that amylase activity correlates with AMY2B copy numbers. We then describe how AMY2B copy numbers vary in individuals from 20 dog breeds and find strong breed-dependent patterns, indicating that the ability to digest starch varies both at the breed and individual level. Finally, to test whether AMY2B copy number is strongly associated with the risk of developing diabetes mellitus, we compare copy numbers in cases and controls as well as in breeds with varying diabetes susceptibility. Although we see no such association here, future studies using larger cohorts are needed before excluding a possible link between AMY2B and diabetes mellitus. PMID:24975239

Arendt, Maja; Fall, Tove; Lindblad-Toh, Kerstin; Axelsson, Erik

2014-10-01

51

Amylase activity and growth in internodes of deepwater rice.  

PubMed

Isoelectrofocusing, product analysis, thermal denaturation studies and affinity chromatography on cycloheptaamylose-Sephadex were used to identify the amylolytic enzymes in internodes of deepwater rice (Oryza sativa L.). Amylolytic activity in internodes of deepwater rice consists of ?-amylase (sometimes separated into two isoforms) and of ?-amylase. During submergence of whole plants, ?-amylase activity increases in young, growing internodes, but ?-amylase activity declines. Although non-growing, mature internodes contain higher levels of ?-amylase than do the elongating younger internodes, the effect of submergence on amylase activities in both tissues follows the same trend. Submergence, gibberellic acid (GA3) and ethylene all promote ?-amylase activity in growing and non-growing internodes of excised deepwater-rice stem sections. Inhibitor studies showed that submergence and ethylene promote ?-amylase activity in the absence of endogenous gibberellin (GA), and GA3 enhances ?-amylase activity when ethylene action is inhibited. Therefore, ethylene and GA appear to increase ?-amylase activity independently of each other. Enhanced ?-amylase activities are probably responsible for the mobilization of carbohydrates which are needed to support internode elongation during submergence of deepwater rice. PMID:24225795

Smith, M A; Jacobsen, J V; Kende, H

1987-09-01

52

Lactase persistence and augmented salivary alpha-amylase gene copy numbers might have been selected by the combined toxic effects of gluten and (food born) pathogens.  

PubMed

Various positively selected adaptations to new nutrients have been identified. Lactase persistence is among the best known, conferring the ability for drinking milk at post weaning age. An augmented number of amylase gene (AMY1) copies, giving rise to higher salivary amylase activity, has been implicated in the consumption of starch-rich foods. Higher AMY1 copy numbers have been demonstrated in populations with recent histories of starchy-rich diets. It is however questionable whether the resulting polymorphisms have exerted positive selection only by providing easily available sources of macro and micronutrients. Humans have explored new environments more than any other animal. Novel environments challenge the host, but especially its immune system with new climatic conditions, food and especially pathogens. With the advent of the agricultural revolution and the concurrent domestication of cattle came new pathogens. We contend that specific new food ingredients (e.g., gluten) and novel pathogens drove selection for lactase persistence and higher AMY gene copy numbers. Both adaptations provide ample glucose for activating the sodium glucose-dependent co-transporter 1 (SGLT1), which is the principal glucose, sodium and water transporter in the gastro-intestinal tract. Their rapid uptake confers protection against potentially lethal dehydration, hyponatremia and ultimately multiple organ failure. Oral rehydration therapy aims at SGLT1 activity and is the current treatment of choice for chronic diarrhoea and vomiting. We hypothesize that lifelong lactase activity and rapid starch digestion should be looked at as the evolutionary covalent of oral rehydration therapy. PMID:24472865

Pruimboom, Leo; Fox, Tom; Muskiet, Frits A J

2014-03-01

53

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...

54

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

PubMed Central

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

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

2013-01-01

55

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

PubMed

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

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

2014-05-01

56

[Alpha-amylase isoenzymes in serum and saliva of patients with anorexia and bulimia nervosa].  

PubMed

In the serum and saliva of 45 patients with eating disorders and in 30 normal controls, alpha-amylase activity and isoamylase levels were measured. Of the 45 patients evaluated, 12 had restrictive anorexia nervosa, 13 were bulimic anorectics and 20 had bulimia nervosa. In all these groups, the mean alpha-amylase values in serum and saliva were higher than that of the control group. The proportion of pancreatic (P)- and salivary (S)-alpha-amylase isoenzymes in serum were within the normal range for the patient group with restrictive anorexia nervosa, whereas the bulimic anorexia nervosa and bulimia nervosa patients showed significantly greater increases in S- than P-isoamylase activity. The correlation of the salivary alpha-Amylase isoenzym pattern in serum and saliva pointed to the salivary glands as origin of the elevated salivary isoamylase levels in serum. Hyperamylasemia was found in 10 (25%) of the 45 patients with eating disorders. Three of these patients showed besides an increased S-alpha-amylase activity also pathologically elevated P-alpha-amylase and lipase activity in serum; however there were no abdominal symptoms, laboratory data or ultrasonic signs of pancreatitis. In all patients with eating disorders, the mean concentration and secretion of alpha-amylase in saliva were increased. Swelling of the salivary glands was present in 14 patients. In these cases the percentage of salivary-isoamylase activity in total serum alpha-amylase activity was increased significantly, whereas the alpha-amylase secretion in the resting saliva was decreased. PMID:1950041

Scheutzel, P; Gerlach, U

1991-07-01

57

Fractionated irradiation and early changes in salivary glands. Different effects on potassium efflux, exocytotic amylase release and gland morphology  

SciTech Connect

Irradiation is a potent treatment modality of head and neck cancer. However, the irradiation is usually associated with an influence on salivary glands with ensuing dryness and discomfort for the patients. In the present study we used different in vitro secretory models and morphologic characterization of rat parotid gland. Radiation was given to one gland on a 5-day schedule with 6 MV photons (total dose 20, 30, 35, 40, 45 Gy). The contralateral gland served as control, and the analysis of glands were performed 10 days after the last irradiation treatment. The noradrenaline stimulated electrolyte secretion (86rubidium tracer for potassium) was decreased in relation to the irradiation dose and in comparison to contralateral control glands. Noradrenaline stimulated exocytotic amylase release was not affected by irradiation and, there were no signs of obvious quantitative morphologic alterations after irradiation compared with controls. The results suggest that there are differences in the sensitivity to radiation for the two different secretory processes in salivary glands, and, thus, the structures regulating electrolyte and fluid secretion seem to be more vulnerable to irradiation than the process of exocytosis. The results, however, do not allow discrimination between temporary cellular impairment and irreversible damage leading to cell death.

Franzen, L.; Funegard, U.S.; Sundstroem, S.G.; Gustafsson, H.; Danielsson, A.; Henriksson, R. (University Hospital, Umea (Sweden))

1991-02-01

58

Thermodynamic stability of a cold-active alpha-amylase from the Antarctic bacterium Alteromonas haloplanctis.  

PubMed

The thermal stability of the cold-active alpha-amylase (AHA) secreted by the Antarctic bacterium Alteromonas haloplanctis has been investigated by intrinsic fluorescence, circular dichroism, and differential scanning calorimetry. It was found that this heat-labile enzyme is the largest known multidomain protein exhibiting a reversible two-state unfolding, as demonstrated by the recovery of DeltaHcal values after consecutive calorimetric transitions, a DeltaHcal/DeltaHeff ratio close to unity, and the independence of unfolding thermodynamic parameters of scan rates. By contrast, the mesophilic alpha-amylases investigated here (from porcine pancreas, human salivary glands, yellow meal beetle, Bacillus amyloliquefaciens, and Bacillus licheniformis) unfold irreversibly according to a non-two-state mechanism. Unlike mesophilic alpha-amylases, the melting point of AHA is independent of calcium and chloride binding while the allosteric and structural functions of these ions are conserved. The thermostability of AHA at optimal conditions is characterized by a Tm of 43.7 degrees C, a DeltaHcal of 238 kcal mol-1, and a DeltaCp of 8.47 kcal mol-1 K-1. These values were used to calculate the Gibbs free energy of unfolding over a wide range of temperatures. This stability curve shows that (a) the specific DeltaGmax of AHA [22 cal (mol of residue)-1] is 4 times lower than that of mesophilic alpha-amylases, (b) group hydration plays a crucial role in the enzyme flexibility at low temperatures, (c) the temperature of cold unfolding closely corresponds to the lower limit of bacterial growth, and (d) the recombinant heat-labile enzyme can be expressed in mesophilic hosts at moderate temperatures. It is also argued that the cold-active alpha-amylase has evolved toward the lowest possible conformational stability of its native state. PMID:10194383

Feller, G; d'Amico, D; Gerday, C

1999-04-01

59

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

PubMed

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

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

2014-09-01

60

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

PubMed Central

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.

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

2014-01-01

61

The functional significance of amylase polymorphism in Drosophila melanogaster VI. Duration of development and amylase activity in larvae when starch is a limiting factor  

Microsoft Academic Search

Two stocks homozygous for the amylase alleles Amy1 and Amy4, 6 were compared for amylase activity during larval development and duration of larval development on a food medium on which addition of starch promotes survival. The two stocks have the same development time when reared under optimal food conditions. Then, the higher amylase activity which characterizes the Amy4, 6 stock

A. J. W. Hoorn; W. Scharloo

1981-01-01

62

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

PubMed

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

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

2013-01-01

63

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

PubMed Central

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

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

2013-01-01

64

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

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…

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

2012-01-01

65

The ram1 Mutant of Arabidopsis Exhibits Severely Decreased ?-Amylase Activity1  

PubMed Central

Despite extensive biochemical analyses, the biological function(s) of plant ?-amylases remains unclear. The fact that ?-amylases degrade starch in vitro suggests that they may play a role in starch metabolism in vivo. ?-Amylases have also been suggested to prevent the accumulation of highly polymerized polysaccharides that might otherwise impede flux through phloem sieve pores. The identification and characterization of a mutant of Arabidopsis var. Columbia with greatly reduced levels of ?-amylase activity is reported here. The reduced ?-amylase 1 (ram1) mutation lies in the gene encoding the major form of ?-amylase in Arabidopsis. Although the Arabidopsis genome contains nine known or putative ?-amylase genes, the fact that the ram1 mutation results in almost complete loss of ?-amylase activity in rosette leaves and inflorescences (stems) indicates that the gene affected by the ram1 mutation is responsible for most of the ?-amylase activity present in these tissues. The leaves of ram1 plants accumulate wild-type levels of starch, soluble sugars, anthocyanin, and chlorophyll. Plants carrying the ram1 mutation also exhibit wild-type rates of phloem exudation and of overall growth. These results suggest that little to no ?-amylase activity is required to maintain normal starch levels, rates of phloem exudation, and overall plant growth. PMID:11743123

Laby, Ron J.; Kim, Donggiun; Gibson, Susan I.

2001-01-01

66

Where does serum amylase come from and where does it go?  

PubMed

The serum amylase concentration reflects the balance between the rates of amylase entry into and removal from the blood. Hyperamylasemia can result either from an increased rate of entry of amylase into the circulation and/or a decreased metabolic clearance of this enzyme. The pancreas and salivary glands have amylase concentrations that are several orders of magnitude greater than that of any other normal tissue, and these two organs probably account for almost all of the serum amylase activity in normal persons. A variety of techniques are now available to distinguish pancreatic from salivary-type isoamylase. Pancreatic hyperamylasemia results from an insult to the pancreas, ranging from trivial (cannulation of the pancreatic duct) to severe (pancreatitis). In addition, loss of bowel integrity (infarction or perforation) causes pancreatic hyperamylasemia due to absorption of amylase from the intestinal lumen. Hyperamylasemia due to salivary-type isoamylase is observed in conditions involving the salivary glands. In addition, this type of hyperamylasemia occurs in conditions in which there is no clinical evidence of salivary gland disease, such as chronic alcoholism, postoperative states (particularly postcoronary bypass), lactic acidosis, anorexia nervosa or bulimia, and malignant neoplasms that secrete amylase. Hyperamylasemia can also result from decreased metabolic clearance of amylase due to renal failure or macroamylasemia (a condition in which an abnormally high-molecular-weight amylase is present in the serum). Patients with abdominal pain and a markedly elevated serum amylase (more than three times the upper limit of normal) usually have acute pancreatitis, and additional serum enzyme testing is not helpful. Patients with smaller elevations of serum amylase often have conditions other than pancreatitis, and measurement of a serum enzyme more specific for the pancreas (pancreatitic isoamylase, lipase or trypsin) is frequently of diagnostic value in such patients. PMID:1702756

Pieper-Bigelow, C; Strocchi, A; Levitt, M D

1990-12-01

67

The effect of sodium hypochlorite treatment on the development of ?-amylase activity in mung bean cotyledons  

Microsoft Academic Search

The development of ?-amylase activity in mung bean cotyledons was severely retarded, when they were detached from the axis. However, when detached cotyledons were treated with sodium hypochlorite (NaOCl), ?-amylase activity was markedly enhanced. The enhancement of the activity was due to a raise in the level of the enzyme protein. The activities of protease and malate dehydrogenase were not

Yasuko Kaneko; Yukio Morohashi

2003-01-01

68

Retroviral and psuedogene insertion sites reveal the lineage of human salivary and pancreatic amylase genes from a single gene during primate evolution  

SciTech Connect

The authors have analyzed the junction regions of inserted elements within the human amylase gene complex. This complex contains five genes which are expressed at high levels either in the pancreas or in the parotid gland. The proximal 5{prime}-flanking regions of these genes contain two inserted elements. A {gamma}-actin pseudogene is located at a position 20 base pairs upstream of the first coding exon. All of the amylase genes contain this insert. The subsequent insertion of an endogenous retrovirus interrupted the {gamma}-actin pseudogene within its 3{prime}-untranslated region. Nucleotide sequence analysis of the inserted elements associated with each of the five human amylase genes has revealed a series of molecular events during the recent history of this gene family. The data indicate that the entire gene family was generated during primate evolution from one ancestral gene copy and that the retroviral insertion activated a cryptic promoter.

Samuelson, L.C.; Snow, C.M.; Meisler, M.H. (Michigan Univ., Ann Arbor, MI (USA). Dept. of Human Genetics); Wiebauer, K. (Friedrich Meischer Inst., CH-4002 Basel (CH))

1990-06-01

69

Potent ?-amylase inhibitory activity of Indian Ayurvedic medicinal plants  

PubMed Central

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

2011-01-01

70

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...

71

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...

72

Production of alpha-amylase by yeast  

SciTech Connect

The enzyme alpha-amylase confers to an organism the enzymatic activity for the degradation of polyglucosides with alpha-1,4 glycosidic bonds such as starch and glycogen which are among the major storage compounds in plants and animals. Most alpha-amylases are single polypeptides of molecular weights around 50,000 dalton. They are generally found in the digestive tract of animals and in germinating seeds. Among the products released upon enzymatic degradation of polyglucosides maltose, a sugar that can be utilized as carbon source by yeast, is a major constituent. A cDNA segment complementary to mouse salivary amylase messenger RNA has been inserted into the yeast expression vector pMA56 behind the promoter of the gene encoding alcohol dehydrogenase I of yeast. Yeast transformants harboring plasmids with the normal orientation of the promoter and the mouse amylase cDNA gene produce amylase and release the enzyme in free form into the culture medium. Approximately 90% of the amylase activity is found in the medium. Yeast strains carrying MAL allele and transformed with a plasmid which directed the synthesis of mouse alpha-amylase were tested on plates containing starch and in batch fermentations using different high molecular weight sugars and oligosaccharides as carbon source. The results of these experiments will be discussed. (Refs. 21).

Thomse, K.K.

1987-01-01

73

Smoking increases salivary arginase activity in patients with dental implants  

Microsoft Academic Search

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

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

2009-01-01

74

Human Salivary Mucin MG1 Selectively Forms Heterotypic Complexes with Amylase, Proline-rich Proteins, Statherin, and Histatins  

Microsoft Academic Search

Heterotypic complexes between the high-molecular-weight mucin MG1 and other salivary proteins in human submandibular\\/sublingual secretion (HSMSL) could have a significant impact on the biological properties of these proteins in oral fluids in both health and disease. We describe a mild procedure for isolation and purification of native MG1 by gel filtration chromatography on Sepharose CL-2B which does not involve dialysis,

I. Iontcheva; F. G. Oppenheim; R. F. Troxler

1997-01-01

75

A 1.2 kb deletion in the 5' region of the beta-amylase gene is responsible for the lack of beta-amylase activity in soybean cultivar Altona sp 1  

Technology Transfer Automated Retrieval System (TEKTRAN)

Previous studies have identified near-isogenic soybean lines, one containing normal beta-amylase activity (Altona Sp 1b) and the other with undetectable beta-amylase activity (Altona sp 1). The molecular basis for the absence of beta-amylase activity in the mutant has not been investigated. In thi...

76

beta-amylase in developing apple fruits: activities, amounts and subcellular localization.  

PubMed

Starch degradation in cells is closely associated with cereal seed germination, photosynthesis in leaves, carbohydrate storage in tuberous roots, and fleshy fruit development. Based on previously reported in vitro assays, beta-amylase is considered one of the key enzymes catalyzing starch breakdown, but up to date its role in starch breakdown in living cells remains unclear because the enzyme was shown often extrachloroplastic in living cells. The present experiment showed that beta-amylase activity was progressively increasing concomitantly with decreasing starch concentrations during apple (Malus domestica Borkh cv. Starkrimson) fruit development. The apparent amount of beta-amylase assessed by Western blotting also increased during the fruit development, which is consistent with the seasonal changes in the enzyme activity. The subcellular-localization studies via immunogold electron-microscopy technique showed that beta-amylase visualized by gold particles was predominantly located in plastids especially at periphery of starch granules, but the gold particles were scarcely found in other subcellular compartments. These data proved for the first time that the enzyme is compartmented in its functional sites in plant living cells. The predominantly plastid-distributed pattern of beta-amylase in cells was shown unchanged throughout the fruit development. The density of gold particles (beta-amylase) in plastids was increasing during the fruit development, which is consistent with the results of Western blotting. So it is considered that beta-amylase is involved in starch hydrolysis in plastids of the fruit cells. PMID:18759031

Zhang, Dapeng; Wang, Yongzhang

2002-08-01

77

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

PubMed Central

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

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

78

Complement activation by salivary agglutinin is secretor status dependent.  

PubMed

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

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

2015-01-01

79

Cholecystokinin receptors: disparity between phosphoinositide breakdown and amylase releasing activity of CCK analogues in pancreas  

SciTech Connect

Cholecystokinin (CCK) peptides are a family of hormones which also occur in brain. In pancreas CCK stimulates the release of amylase, a process that is dependent on the mobilization of intracellular Ca/sup 2 +/. Recent evidence suggests that inositol 1,4,5-trisphosphate, the breakdown product of phosphatidylinositol 4,5-bisphosphate, is responsible for the rise in intracellular Ca/sup 2 +/. Their laboratory has developed assays to study synthetic CCK analogues using radioligand binding, PI breakdown and amylase release. They have shown that there are good correlations among these three assay systems for the carboxy terminal fragments of CCK/sub 8/. Recently, they have discovered synthetic analogues of CCK/sub 4/ that are full agonists in amylase release but are ineffective in causing PI breakdown. In particular, A-61576, Boc-5-amino-2-indolemethylene-pent-2-ene-1-oyl-Leu-Asp-Phe-NH/sub 2/, is a full agonist in the amylase releasing assay, but is devoid of PI stimulating activity. A-61576 completely reverses the stimulation of PI response induced by CCK/sub 8/, indicative of an antagonist. Since a mechanism other than the PI breakdown is responsible for amylase release by A-61576, they suggest that separate receptors are responsible for PI breakdown and amylase release.

Lin, C.W.; Grant, D.; Bianchi, B.; Miller, T.; Witte, D.; Shue, Y.K.; Nadzan, A.

1986-03-05

80

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

PubMed

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

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

2014-05-01

81

Comparative study of digestive enzymes in fish with different nutritional habits. Proteolytic and amylase activities  

Microsoft Academic Search

This work provides a comparative study of the proteolytic and amylase activities in six species of fish with different nutritional habits: rainbow trout (Oncorhynchus mykiss), gilthead seabream (Sparus aurata), European eel (Anguilla anguilla), common carp (Cyprinus carpio), goldfish (Carassius auratus), and tench (Tinca tinca). Trout and carp showed the highest digestive proteolytic activity. When proteolytic activity was determined in a

M. C Hidalgo; E Urea; A Sanz

1999-01-01

82

Amylase Test  

MedlinePLUS

... of this website will be limited. Search Help? Amylase Share this page: Was this page helpful? Also ... between P-amylase and S-amylase? 1. Do elevated amylase levels always mean that I have a pancreatic ...

83

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...

84

Preparation, characterization and biocatalytic activity of a nanoconjugate of alpha amylase and silver nanoparticles.  

PubMed

The primary challenge in developing nanoparticle based enzymatic devices is to be able to chemically immobilize an enzyme, which will retain its activity or improve its function while being attached to the nanoparticle. This would be of even greater significance if the whole process could be performed under benign conditions without having to resort to functionalization of key molecules at various steps. In the present study the conjugates of amylase and silver nanoparticles were synthesized using neem leaf extract as the reducing and stabilizing agent. The silver nanoparticles were characterized using Surface Plasmon Resonance Spectra, Dynamic Light Spectroscopy (DLS), Fourier Transform Infrared Spectroscopy (FTIR), Circular Dichroism (CD) and Surface Tunneling Microscopy (STM). The silver nanoparticles retained 85% amylase activity. The nanobiocatalyst was further characterized in terms of kinetic parameters and thermal stability. It was thermally more stable as compared to the free alpha amylase enzyme. PMID:23901526

Mishra, Abhijeet; Ahmad, Razi; Singh, Veena; Gupta, Munishwar Nath; Sardar, Meryam

2013-07-01

85

Screening of Mongolian plants for influence on amylase activity in mouse plasma and gastrointestinal tube.  

PubMed

Mongolian plants were screened for their influence on alpha-amylase activity in mouse plasma. Methanolic extracts of Geranium pratense, Rhodiola rosea, Ribes pullchelum and Vaccinium uliginosum inhibited the enzyme activity in isolated mouse plasma by greater than 40% and the effect was concentration dependent. Vaccinium uliginosum also showed a depressive effect on elevation of postprandial blood glucose to some extent. PMID:12843638

Kobayashi, Kyoko; Baba, Eriko; Fushiya, Shinji; Takano, Fumihide; Batkhuu, Javzan; Dash, Tzezengiin; Sanchir, Chinbat; Yoshizaki, Fumihiko

2003-07-01

86

In Vitro ?-Amylase Inhibition and Antioxidant Activities of Methanolic Extract of Amaranthus Caudatus Linn  

PubMed Central

Objectives The present study was aimed to investigate the ?-amylase inhibition and antioxidant activities of methanolic extract of Amaranthus caudatus Linn (MeAc). Methods Methanolic extract of Amaranthus caudatus was screened for ?-amylase inhibition activity by CNPG3 method (2-chloro-p-nitrophenyl-?-D-maltotrioside) and antioxidant activity was evaluated by 1,1-diphenyl-2-picryl-hydrazile (DPPH) free radical scavenging, superoxide dismutase (SOD) scavenging, hydroxyl free radical scavenging, nitric oxide (NO) radical scavenging, and 2.2’-azinobis-3-ethylbenzothiazole-6-sulfonic acid (ABTS) radical scavenging assays. MeAc was also screened for non enzymatic hemoglycosylation. Results The methanolic extract of Amaranthus caudatus showed potent ?-amylase inhibition activity (IC50 19.233 µg/ml). MeAc showed significant antioxidant activity in all the in vitro antioxidant models. Furthermore, the MeAc was found to be extremely effective in scavenging ABTS radical activity (IC50 48.75±1.1 µg/ml) when compared to DPPH (IC50 77.5±0.4 µg/ml), SOD (IC50 62.5±2.1 µg/ml), hydroxyl (IC50 88.50±1.8 µg/ml) and NO (IC50 67.5±2.2 µg/ml) scavenging activity. Conclusions The methanolic extract of A. caudatus showed potent ?-amylase inhibition and antioxidant activities. PMID:22043408

Kumar, Ashok; Khan, Saleemulla

2011-01-01

87

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

PubMed

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

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

2015-03-01

88

A novel method to estimate changes in stress-induced salivary ?-amylase using heart rate variability and respiratory rate, as measured in a non-contact manner using a single radar attached to the back of a chair.  

PubMed

The authors have developed a non-contact system which estimates changes in salivary ?-amylase (sAA ratio) induced by stress. Before and after stressful sound exposure, a single 24?GHz compact radar which is attached to the back of a chair measures the low frequency (LF) component of heart rate variability and respiratory rate, ?-amylase in the subjects' buccal secretions was measured by using an ?-amylase assay kit. Using multiple regression analysis, sAA ratio was estimated using stress-induced LF change (LF ratio) and stress-induced respiratory rate change (respiratory rate ratio). Twelve healthy subjects were tested (12 males, 22?±?2 years), who were exposed to audio stimuli with a composite tone of 2120?Hz and 2130?Hz sine waves at a sound pressure level of 95?dB after a silent period through a headphone. The result showed that sAA ratio estimated using multiple regression analysis significantly correlated with measured sAA ratio (R?=?0.76, p?

Matsui, Takemi; Katayose, Satoshi

2014-08-01

89

Development of an industrial method to quantitatively measure carry-over amylase activity in raw and refined sugars  

Technology Transfer Automated Retrieval System (TEKTRAN)

In recent years, there has been increased concern over carry-over activity of mostly high temperature (HT) and very high temperature (VHT) stable amylases in white, refined sugars from refineries to various food manufacturing industries and other end-users. HT and VHT stable amylases were developed...

90

The need for and development of a method to measure carry-over amylase activity in raw and refined sugars  

Technology Transfer Automated Retrieval System (TEKTRAN)

In recent years, there has been increased world-wide concern over carry-over activity of mostly high temperature (HT) and very high temperature (VHT) stable amylases in refined sugars to various food and end-user industries. HT and VHT stable amylases were developed for much larger markets than the...

91

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...

92

Structure-based protein engineering for alpha-amylase inhibitory activity of plant defensin.  

PubMed

The structure of a novel plant defensin isolated from the seeds of the mung bean, Vigna radiate, has been determined by (1)H nuclear magnetic resonance spectroscopy. The three-dimensional structure of VrD2, the V. radiate plant defensin 2 protein, comprises an alpha-helix and one triple-stranded anti-parallel beta-sheet stabilized by four disulfide bonds. This protein exhibits neither insecticidal activity nor alpha-amylase inhibitory activity in spite of showing a similar global fold to that of VrD1, an insecticidal plant defensin that has been suggested to function by inhibiting insect alpha-amylase. Our previous study proposed that loop L3 of plant defensins is important for this inhibition. Structural analyses and surface charge comparisons of VrD1 and VrD2 revealed that the charged residues of L3 correlate with the observed difference in inhibitory activities of these proteins. A VrD2 chimera that was produced by transferring the proposed functional loop of VrD1 onto the structurally equivalent loop of VrD2 supported this hypothesis. The VrD2 chimera, which differs by only five residues compared with VrD2, showed obvious activity against Tenebrio molitor alpha-amylase. These results clarify the mode of alpha-amylase inhibition of plant defensins and also represent a possible approach for engineering novel alpha-amylase inhibitors. Plant defensins are important constituents of the innate immune system of plants, and thus the application of protein engineering to this protein family may provide an efficient method for protecting against crop losses. PMID:17444520

Lin, Ku-Feng; Lee, Tian-Ren; Tsai, Ping-Hsing; Hsu, Ming-Pin; Chen, Ching-San; Lyu, Ping-Chiang

2007-08-01

93

The Effects of MMP Inhibitors on Human Salivary MMP Activity and Caries Progression in Rats  

Microsoft Academic Search

Previous studies suggest that salivary and pulp-derived host enzymes, matrix metalloproteinases (MMPs), may be involved in dentin caries pathogenesis. To study the inhibition of acid-activated human salivary MMPs by non-antimicrobial chemically modified tetracyclines (CMTs), we used a functional activity assay with 125I-labeled gelatin as a substrate. To address the role of MMPs in the progression of fissure caries in vivo,

M. Sulkala; J. Wahlgren; M. Larmas; T. Sorsa; O. Teronen; T. Salo; L. Tjäderhane

2001-01-01

94

Amylase - urine  

MedlinePLUS

... is a test that measures the amount of amylase in urine. Amylase is an enzyme that helps digest carbohydrates. It ... the pancreas and the glands that make saliva. Amylase may also be measured with a blood test .

95

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

PubMed Central

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

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

2011-01-01

96

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

PubMed

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

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

2012-09-12

97

False-positive results with amylase testing of citrus fruits.  

PubMed

In a case of robbery in which the criminals passed through the garden adorned with calamondin trees (Citrus madurensis), the investigators found in the grass six calamondin fruits, some undamaged, while others apparently bitten. The fruits were collected and sent to the laboratory for DNA analysis to verify the presence of saliva and robbers' DNA profile. A specific immunochromatographic strip test for saliva confirmed the presence of human salivary ?-amylase, but similar positive results were also observed for intact calamondin and other citrus fruits. Further analysis with a specific automated amylase test confirmed the absence of amylase activity. DNA quantification and typing using a specific forensic kit revealed no human DNA presence in any fruits. This case report demonstrates for the first time the occurrence of false positives when human saliva is sought on citrus fruits. PMID:24502328

Ricci, Ugo; Carboni, Ilaria; Torricelli, Francesca

2014-09-01

98

Estimation of salivary ?-glucuronidase activity as a marker of periodontal disease: A case control study  

PubMed Central

Aim: The aim of the present study was to estimate the salivary ?-glucuronidase level in healthy and diseased periodontium and to correlate the level with clinical measurement. Materials and Methods: 70 patients were included in this study with the age ranging from 30 to 65 years. Both males and females were included. They were divided into two groups: Control having healthy periodontium (n = 20) and experimental having diseased periodontium (n = 50). The parameters recorded were probing pocket depth, probing attachment level, gingival index, ?-glucuronidase activity in the saliva, number of white blood cells, neutrophils, lymphocytes count, and platelet count. Results: It was observed that there was an increase in the level of salivary ?-glucuronidase in the experimental subjects than in the control patients, and a significant positive linear relationship existed between salivary ?-glucuronidase level and probing pocket depth in the experimental group. Conclusion: Level of salivary ?-glucuronidase increases during inflammation in the periodontium.

Prabhahar, Chandra Sekhara; Niazi, K. Thanvir Mohamed; Prakash, R.; Yuvaraj, A.; Goud, Somasekhar; Ravishekar, P.

2014-01-01

99

Effects of metals on {alpha}-amylase activity in the digestive gland of the green mussel, Perna viridis L.  

SciTech Connect

A number of digestive enzymes in the green mussel, Perna viridis L., have been reported, and {alpha}-amylase is believed to have a higher activity than the others. Small plankton, on which the green mussel feeds, may supply plenty of starch and glycogen. They may be an important source of nutrients for the green mussel and the ability of the latter to make good use of them depends mainly on the activities of amylase. The effect of heavy metals on amylase activity is also important as the ability of the mussel`s digestive gland to accumulate these metals is well known. High concentrations of heavy metals, especially lead, have been observed in the water around Singapore. The in vitro inhibition of some metals on the activities of digestive enzymes from the green mussel has been observed, but kinetic properties of the inhibition and the in vivo inhibition of the heavy metals on digestive enzymes are little understood. In the present study, in vitro inhibition of four metals (Pb, Cd, Zn and Hg) on the activity of {alpha}-amylase from the digestive gland of the green mussel will be compared. Their effects on the K{sub M} and V{sub max} values of {alpha}-amylase will also be compared. Finally, lead is either added to the food or water, to see how it affects the activity of {alpha}-amylase and how this effect acts in combination with starvation. 12 refs., 3 figs., 3 tabs.

Yan, T.; Teo, L.H.; Sin, Y.M. [National Univ. of Singapore (Singapore)] [National Univ. of Singapore (Singapore)

1996-04-01

100

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

PubMed

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

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

101

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.  

PubMed

This article presents two integrated laboratory exercises intended to show students the role of ?-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 (qualitative) under different conditions (e.g. variations in temperature and alkalinity). This work also proposes the study of enzyme stability in the presence of several surfactants and oxidizing agents using the same technical approach. The proposed laboratory exercises promote the understanding of the physiological function of this enzyme and the biotechnological applications of AAMYs in the detergent industry. The exercises also promote the understanding that the enzymatic stability and performance are dependent on the organism of origin, and if necessary, these properties could be modified by genetic engineering. In addition, this article reinforces the development of laboratory skills, problem-solving capabilities, and the ability to write a laboratory report. The exercises are proposed primarily as an undergraduate project for advanced students in the biochemical and biotechnological sciences. These laboratory practicals are complementary to the previously published BAMBED article (Biochemistry and Molecular Biology Education Vol. 39, No. 4, pp. 280-290, 2011) on detergent proteases. PMID:22807429

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

2012-07-01

102

Using a Short Wavelength Infrared (SWIR) hyperspectral imaging system to predict alpha amylase activity in individual Canadian western wheat kernels  

Microsoft Academic Search

Sprout damage (pre-harvest germination) in wheat results in highly deleterious effects on end-product quality. Alpha-amylase,\\u000a the pre-dominant enzyme in the early stage of sprouting has the most damaging effect. This paper introduces a new method using\\u000a a SWIR hyperspectral imaging system (1000–2500 nm) to predict the ?-amylase activity of individual wheat kernels. Two classes\\u000a of Canadian wheat, Canada Western Red Spring

Juan Xing; Pham Van Hung; Stephen Symons; Muhammad Shahin; David Hatcher

2009-01-01

103

Concerted evolution of human amylase genes  

SciTech Connect

Cosmid clones containing 250 kilobases of genomic DNA from the human amylase gene cluster have been isolated. These clones contain seven distinct amylase genes which appear to comprise the complete multigene family. By sequence comparison with the cDNAs, the authors have identified two pancreatic amylase gene and three salivary amylase genes. Two truncated pseudogenes were also recovered. Intergenic distances of 17 to 22 kilobases separate the amylase gene copies. Within the past 10 million years, duplications, gene conversion, and unequal crossover events have resulted in a very high level of sequence similarity among human amylase gene copies. To identify sequence elements involved in tissue-specific expression and hormonal regulation, the promoter regions of the human amylase genes were sequenced and compared with those of the corresponding mouse genes. The promoters of the human and mouse pancreatic amylase genes are highly homologous between nucleotide - 160 and the cap site. Two sequence elements througth to influence pancreas-specific expression of the rodent genes are present in the human genes. In contrast, similarity in the 5' lanking sequences of the salivary amylase genes is limited to several short sequence elements whose positions and orientations differ in the two species. Some of these sequence elements are also associated with other parotid-specific genes and may be involved in their tissue-specific expression. A glucocorticoid response element and a general enhancer element are closely associated in several of the amylase promoters.

Gumucio, D.L.; Wiebauer, K.; Caldwell, R.M.; Samuelson, L.C.; Meisler, M.H.

1988-03-01

104

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

PubMed

?-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 10MHz. 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 (0.08-8U/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

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

2014-09-21

105

Activation of bean (Phaseolus vulgaris) [alpha]-amylase inhibitor requires proteolytic processing of the proprotein  

SciTech Connect

Seeds of the common bean (Phaseolus vulgaris) contain a plant defense protein that inhibits the [alpha]-amylases of mammals and insects. This [alpha]-amylase inhibitor ([alpha]Al) is synthesized as a proprotein on the endoplasmic reticulum and is proteolytically processed after arrival in the protein storage vacuoles to polypeptides of relative molecular weight (M[sub r]) 15,000 to 18,000. The authors report two types of evidence that proteolytic processing is linked to activation of the inhibitory activity. First, by surveying seed extracts of wild accessions of P. vulgaris and other species in the genus Phaseolus, they found that antibodies to [alpha]Al recognize large (M[sub r] 30,000-35,000) polypeptides as well as typical [alpha]Al processing products (M[sub r] 15,000-18,000). [alpha]Al activity was found in all extracts that had the typical [alpha]Al processed polypeptides, but was absent from seed extracts that lacked such polypeptides. Second, they made a mutant [alpha]Al in which asparagine-77 is changed to aspartic acid-77. This mutation slows down the proteolytic processing of pro-[alpha]Al when the gene is expressed in tobacco. When pro-[alpha]Al was separated from mature [alpha]Al by gel filtration, pro-[alpha]Al was found not to have [alpha]-amylase inhibitory activity. The authors interpret these results to mean that formation of the active inhibitor is causally related to proteolytic processing of the proprotein. They suggest that the polypeptide cleavage removes a conformation constraint on the precursor to produce the biochemically active molecule. 43 refs., 5 figs., 1 tab.

Pueyo, J.J.; Hunt, D.C.; Chrispeels, M.J. (Univ. of California, San Diego, La Jolla (United States))

1993-04-01

106

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

PubMed Central

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

Kumar, Sumit; Khare, S. K.

2015-01-01

107

Determination of antioxidant capacity, ?-amylase and lipase inhibitory activity of Crotalaria juncea Linn in vitro inhibitory activity of Crotalaria Juncea Linn.  

PubMed

The present study involves the determination of antioxidant capacity and in vitro ?-amylase and lipase inhibitory activity of the Crotalaria juncea Linn extract. The content of polyphenols, flavonoids, and tannins in the extracts was estimated by spectrophotometry. Antioxidant activity on goat liver lipid peroxidation and linoleic acid emulsion were determined and ?-amylase and lipase inhibitory activity was also evaluated. All the extracts had shown antioxidant property, ?-amylase, and lipase inhibitory properties. Aqueous extract was found to show maximum antioxidant activity on goat liver. Antilipid peroxidation and antioxidant activity were determined to be 66.94 ± 0.616 (p < .01) and 59.54 ± 0.2 (p < .01), respectively. Maximum ?-amylase and lipase inhibitory activities of 71.42 ± 1.37 (p < .01) and 57.14 ± 2.74% (p < .01), respectively, were exhibited by macerated methanol extract. The results had shown that all the extracts exhibited low inhibition and antioxidant activity as compared to standard. PMID:24670121

Dinakaran, Sathis Kumar; Banji, David; Avasarala, Harani; Banji, Otilia

2014-06-01

108

?-amylase from wheat (Triticum aestivum) seeds: its purification, biochemical attributes and active site studies.  

PubMed

Glycosylated ?-amylase from germinated wheat seeds (Triticum aestivum) has been purified to apparent electrophoretic homogeneity with a final specific activity of 1,372 U/mg. The enzyme preparation when analysed on SDS-PAGE, displayed a single protein band with Mr 33 kDa; Superdex 200 column showed Mr of 32 kDa and MS/MS analysis further provided support for these values. The enzyme displayed its optimum catalytic activity at pH 5.0 and 68 °C with an activation energy of 6.66 kcal/mol and Q10 1.42. The primary substrate for this hydrolase appears to be starch with Km 1.56 mg/mL, Vmax 1666.67 U/mg and kcat 485 s(-1) and hence is suitable for application in starch based industries. Thermal inactivation of ?-amylase at 67 °C resulted in first-order kinetics with rate constant (k) 0.0086 min(-1) and t1/2 80 min. The enzyme was susceptible to EDTA (10mM) with irreversible loss of hydrolytic power. In the presence of 1.0mM SDS, the enzyme lost only 14% and 23% activity in 24 and 48 h, respectively. Chemical modification studies showed that the enzyme contains histidine and carboxylic residues at its active site for its catalytic activity and possibly conserved areas. PMID:24874349

Singh, Kritika; Kayastha, Arvind M

2014-11-01

109

Amylase - blood  

MedlinePLUS

Amylase is an enzyme that helps digest carbohydrates. It is produced in the pancreas and the glands ... saliva. When the pancreas is diseased or inflamed, amylase releases into the blood. A test can be ...

110

Anticoagulation activity of salivary gland extract of oriental blackfly Simulium indicum  

PubMed Central

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

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

2014-01-01

111

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

PubMed

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

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

2014-07-01

112

Comparison of Salivary Beta Glucuronidase Activity in Chronic Periodontitis Patients with and without Diabetes Mellitus  

PubMed Central

Aim of the study: The aim of the study was to estimate the salivary beta glucuronidase (?) activity in patients with chronic periodontitis with and without diabetes mellitus and to evaluate the relationship between Beta Glucuronidase activity and Periodontal clinical parameters. Materials and Methods: The study consisted of 80 patients of both sexes with age ranging from 20-60 years and they were divided into four groups. Clinical parameters such as Gingival index, Probing depth and Clinical attachment loss were measured. Salivary Beta Glucuronidase activity was measured using spectrophotometer with reagents like phenolphthalein glucuronic acid, phosphate and glycine buffer. Results: The mean BG activity of Group IV (1.17 ± 0.27) was significantly higher than mean BGA levels of Group I, II, III. The p-value was < 0.05. The mean BGA levels of Group III (0.78 ± 0.17) was significantly higher than mean BGA levels of Group I, Group II at 5 % level. There was a significant positive linear relationship between salivary ? Glucuronidase level and Probing Depth, clinical attachment level in the experimental Groups. Conclusion: The salivary ? Glucuronidase level was higher in Diabetic patients with periodontitis than nondiabetic periodontitis patients. PMID:25121058

ND, Jayakumar; Varghese, Sheeja

2014-01-01

113

Vampire Bat Salivary Plasminogen Activator (Desmoteplase) A Unique Fibrinolytic Enzyme That Does Not Promote Neurodegeneration  

Microsoft Academic Search

Background and Purpose—Tissue-type plasminogen activator (tPA) promotes excitotoxic and ischemic injury within the brain. These findings have implications for the use of tPA in the treatment of acute ischemic stroke. The plasminogen activator from vampire bat (Desmodus rotundus) saliva (D rotundus salivary plasminogen activator (DSPA); desmoteplase) is an effective plasminogen activator but, in contrast to tPA, is nearly inactive in

Gabriel T. Liberatore; André Samson; Christopher Bladin; Wolf-Dieter Schleuning; Robert L. Medcalf

114

INVITRO EVALUATION OF ?-AMYLASE INHIBITORY ACTIVITY & ANTIOXIDANT POTENTIAL OF PTERIS VITTATA L. WITH SPECIAL REFERENCE TO ITS HPTLC PROFILE.  

E-print Network

Pteris vittata L., a fern was evaluated for its alpha amylase inhibitory potential owing to its previously reported antihyperglycemic activity. This Pteridophyte was also assayed for its free radical inhibition ability in ABTS decolourisation assay followed by quantification and HPTLC of polyphenols. The Porcine pancreatic alpha amylase inhibition was studied invitro and compared with standard drug Acarbose and their IC50 values were determined. For Antioxidant Potential, the ABTS free Radical Scavenging activity was performed and concentration of the test extracts equivalent to ascorbic acid was determined. The phenols and flavonoids being held responsible for antioxidant propertiesof plants lead us to quantify them which were followed by HPTLC fingerprinting of flavonoids to facilitate its identification. Standard Quercetin was used as standard along with the two extracts of Pteris vittata L. As compared to aqueous extract, the ethanolic extract showed better results in the parameters undertaken to study. KEYWORDS:Pteris vittata L., Alpha amylase inhibition, ABTS,Phenol, Flavonoid,HPTLC.

Tania Paul; Suchitra Banerjee; Tania Paul

115

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

PubMed Central

?-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

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

2014-01-01

116

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

PubMed Central

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

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

2014-01-01

117

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

PubMed

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

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

2014-09-01

118

Immunohistochemistry of Active Gibberellins and Gibberellin-Inducible ?-Amylase in Developing Seeds of Morning Glory1  

PubMed Central

Gibberellins (GAs) in developing seeds of morning glory (Pharbitis nil) were quantified and localized by immunostaining. The starch grains began to be digested after the GA contents had increased and reached a plateau. Immunohistochemical staining with the antigibberellin A1-methyl ester-antiserum, which has high affinity to biologically active GAs, showed that GA1 and/or GA3 were localized around starch grains in the integument of developing young seeds, suggesting the participation of GA-inducible ?-amylase in this digestion. We isolated an ?-amylase cDNA (PnAmy1) that was expressed in the immature seeds, and using an antibody raised against recombinant protein, it was shown that PnAmy1 was expressed in the immature seeds. GA responsiveness of PnAmy1 was shown by treating the young fruits 9 d after anthesis with GA3. RNA-blot and immunoblot analyses showed that PnAmy1 emerged soon after the rapid increase of GA1/3. An immunohistochemical analysis of PnAmy1 showed that it, like the seed GA1/3, was also localized around starch grains in the integument of developing young seeds. The localization of GA1/3 in the integument coincident with the expression of PnAmy1 suggests that both function as part of a process to release sugars for translocation or for the further development of the seeds. PMID:12114559

Nakayama, Akira; Park, Seijin; Zheng-Jun, Xu; Nakajima, Masatoshi; Yamaguchi, Isomaro

2002-01-01

119

Insecticidal activity of an alpha-amylase inhibitor-like protein resembling a putative precursor of alpha-amylase inhibitor in the common bean, Phaseolus vulgaris L.  

PubMed

alpha-Amylase inhibitor (alphaAI) in the common bean, Phaseolus vulgaris L., protects seeds from insect pests such as the cowpea weevil (Callosobruchus maculatus) and the azuki bean weevil (C. chinensis). Cultivars which lack alphaAI still show resistance to both bruchids. These cultivars have a glycoprotein that reacts with anti-alphaAI-1 antibodies. The glycoprotein with a molecular mass of 29 kDa (Gp29) was purified and the encoding gene was isolated. The primary structure of Gp29 is the same as alpha-amylase inhibitor-like protein (AIL) from which the encoding gene has already been isolated. AIL resembles a putative precursor of alphaAI, even though it does not form the active inhibitor. However, AIL has some inhibitory effect on the growth of C. maculatus but not C. chinensis. The presence of AIL alone is insufficient to explain the bruchid resistance of common bean cultivars lacking alpha-AI. Common bean seeds appear to contain several factors responsible for the bruchid resistance. PMID:10366733

Ishimoto, M; Yamada, T; Kaga, A

1999-06-15

120

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 ...

121

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...

122

Effects of different food commodities on larval development and ?-amylase activity of Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae)  

Microsoft Academic Search

The present work was undertaken to study the influence of four commodities (wheat flour, dates, sorghum and barley) on Plodia interpunctella post-embryonic development. Larval weight, larval mortality, pupation and adult emergence were recorded. The study also aimed to find out the effect of these commodities on protein and glycogen production as well as on ?-amylase activity. Results indicated that the

Noureddin Bouayad; Kacem Rharrabe; Naima Ghailani; Fouad Sayah

2008-01-01

123

Relationship between malt qualities and ?-amylase activity and protein content as affected by timing of nitrogen fertilizer application*  

PubMed Central

The effects of different timing of N fertilizer application at the same rate on grain ?-amylase activity, protein concentration, weight and malt quality of barley were studied. Grain ?-amylase activity and protein concentration were significantly higher in treatments where all top-dressed N fertilizer was applied at booting stage only or equally applied at two-leaf stage and booting stage than in the treatment where all top-dressed N fertilizer was applied at two-leaf age stage only. On the other hand, grain weight and malt extract decreased with increased N application at booting stage. There were obvious differences between barley varieties and experimental years in the grain and malt quality response to the timing of N fertilizer application. It was found that grain protein concentration was significantly and positively correlated with ?-amylase activity, but significantly and negatively correlated with malt extract and Kolbach index. The effect of grain protein concentration on malt quality was predominant over the effect of grain ?-amylase activity. PMID:16365930

Chen, Jin-xin; Dai, Fei; Wei, Kang; Zhang, Guo-ping

2006-01-01

124

Diet and the evolution of human amylase gene copy number variation  

E-print Network

Diet and the evolution of human amylase gene copy number variation George H Perry1,2, Nathaniel J­3. This behavioral variation raises the possibility that different selective pressures have acted on amylase, the enzyme responsible for starch hydrolysis4. We found that copy number of the salivary amylase gene (AMY1

Sorenson, Michael

125

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

PubMed

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

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

2014-08-01

126

An experimental study of the job demand-control model with measures of heart rate variability and salivary alpha-amylase: Evidence of increased stress responses to increased break autonomy.  

PubMed

We assessed in an experimental design whether the stress response towards a work task was moderated by the autonomy to choose a break during the assigned time to complete the task. This setting is defined in accordance with the theoretical framework of the job-demand-control (JDC) model of work related stress. The findings from naturalistic investigations of a stress-buffering effect of autonomy (or 'buffer hypothesis') are equivocal and the experimental evidence is limited, especially with relation to physiological indices of stress. Our objective was to investigate if increased autonomy in a particular domain (break time control) was related with adaptive physiology using objective physiological markers of stress; heart rate variability (HRV) and salivary alpha amylase (sAA). We used a within-subject design and the 60 female participants were randomly assigned to an autonomy (free timing of break) and standard conditions (fixed timing of break) of a word processing task in a simulated office environment in a random order. Participants reported increased perceptions of autonomy, no difference in demand and performed worse in the task in the break-time autonomy versus the standard condition. The results revealed support for the manipulation of increased autonomy, but in the opposing direction. Increased autonomy was related with dysregulated physiological reactivity, synonymous with typical increased stress responses. Potentially, our findings may indicate that autonomy is not necessary a resource but could become an additional stressor when it adds additional complexity while the amount of work (demands) remains unchanged. Further, our findings underscore the need to collect objective physiological evidence of stress to supplement self-reported information. Self-report biases may partially explain the inconsistent findings with the buffer hypothesis. PMID:25290345

O'Donnell, Emma; Landolt, Kathleen; Hazi, Agnes; Dragano, Nico; Wright, Bradley J

2015-01-01

127

Impaired Histatin-5 Levels and Salivary Antimicrobial Activity against C. albicans in HIV Infected Individuals  

PubMed Central

HIV-infected individuals constitute a population highly susceptible to opportunistic infections, particularly oral candidiasis caused by the most pathogenic human fungal species Candida albicans. Host-produced salivary antimicrobial peptides are considered to be an important part of the host innate immune system involved in protection of the oral cavity against colonization and infection by microbial species. Histatin-5 (Hst-5) specifically has exhibited potent anti-candidal properties in vitro. However, its importance in protecting the oral mucosa against candidal colonization and importantly, its contribution to the observed enhanced susceptibility of HIV-infected individuals to candidiasis has not been previously investigated. To that end, a novel immunoassay was used to demonstrate significant decrease in salivary Hst-5 levels in HIV+ individuals concomitant with enhanced candidal prevalence. Further, saliva’s anti-candidal potency was found to be proportional to Hst-5 concentration and significantly compromised in HIV+ subjects compared to controls. The key role for Hst-5 was further confirmed upon exposure to the Hst-5-specific antibody where saliva’s initial killing activity was substantially compromised. Combined, these findings identify Hst-5 as a key anti-candidal salivary component and demonstrate its decreased levels in HIV infection providing new insights into oral Innate immune defense mechanisms and the enhanced susceptibility of HIV+ individuals to oral candidiasis. PMID:23730535

Khan, Shariq A; Fidel, Paul L; Thunayyan, Awdah Al; Varlotta, Sharon; Meiller, Timothy F; Jabra-Rizk, Mary Ann

2013-01-01

128

Salivary soluble receptor activator of nuclear factor kappa B ligand/osteoprotegerin ratio in periodontal disease and health  

PubMed Central

Purpose The receptor activator of nuclear factor kappa B (RANK)/RANK ligand (RANKL)/osteoprotegerin (OPG) system plays a significant role in osteoclastogenesis, activation of osteoclasts, and regulation of bone resorption. This study aimed to evaluate the use of the salivary soluble RANKL (sRANKL)/OPG ratio as a diagnostic marker for periodontitis in nonsmokers. Methods Twenty-five patients with chronic periodontitis and 25 individuals with a healthy periodontium were enrolled in this study. Samples containing 5 mL of unstimulated saliva were obtained from each subject. Salivary sRANKL and OPG concentrations were determined using a standard enzyme-linked immunosorbent assay. Statistical analysis was performed using SPSS ver. 18.0. Results The levels of sRANKL and OPG were detectable in all of the samples. Positive relationships were found between the plaque index and clinical attachment level and both the salivary concentration of sRANKL and the salivary sRANKL/OPG ratio (P<0.05). The salivary concentration of sRANKL and the sRANKL/OPG ratio were significantly higher in the periodontitis group than in the healthy group (P=0.004 and P=0.001, respectively). In contrast, the OPG concentration showed no significant differences between the groups (P=0.455). Conclusions These findings suggest that the salivary sRANKL/OPG ratio may be helpful in the screening and diagnosis of periodontitis. However, longitudinal studies with larger populations are needed to confirm these results. PMID:24236245

Tabari, Zahra Alizadeh; Azadmehr, Abbas; Tabrizi, Mohammad Amir Alizadeh; Hamissi, Jalaloddin

2013-01-01

129

In Vitro???-Amylase Inhibitory Activity of the Leaves of Thespesia populnea  

PubMed Central

Postprandial hyperglycemia is a prime characteristic of diabetes mellitus and has been a focus in the therapy for diabetes. One of the therapeutic approaches which involve decreasing hyperglycemia aims at inhibiting the enzyme ?-amylase. The leaves of T. populnea were studied for the presence of amylase inhibitors. The fractions obtained by successive fractionation using solvents of varying polarity were studied for the presence of primary and secondary metabolites. The total phenolic content of the different fractions was determined by HPLC and was correlated with their amylase inhibitory potential. Similarly, the protein content of the extracts was also estimated to understand the nature of the inhibitor present. This study shows that the leaves of T. populnea were effective in inhibiting ?-amylase, thereby proving to be potential hyperglycemic agents. PMID:22550597

Sangeetha, R.; Vedasree, N.

2012-01-01

130

Site-directed mutagenesis of putative active-site residues of Bacillus stearothermophilus ?-amylase  

Microsoft Academic Search

Putative catalytic residues of the thermostable Bacillus stearothermophilus a-amylase derived by sequence analysis and computer modeling were tested by site-directed mutagenesis. The conservative mutations produced were Asp-234-Glu, Glu-264-Asp, and Asp-331-Asn. The corresponding amino acids have been proposed to act in acid-base catalysis in the Aspergillus oryzae and porcine pancreatic a-amylase. Isoelectric focusing and immunodiffusion studies showed that, although inactive, the

Mauno Vihinen; Sari Helin; Pekka Mäntsälä

1991-01-01

131

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

PubMed

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

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

2015-03-01

132

Vampire Bat Salivary Plasminogen Activator (Desmoteplase) Inhibits Tissue-Type Plasminogen Activator-Induced Potentiation of Excitotoxic Injury  

Microsoft Academic Search

Background and Purpose—In contrast to tissue-type plasminogen activator (tPA), vampire bat (Desmodus rotundus) salivary plasminogen activator (desmoteplase (DSPA)) does not promote excitotoxic injury when injected directly into the brain. We have compared the excitotoxic effects of intravenously delivered tPA and DSPA and determined whether DSPA can antagonize the neurotoxic and calcium enhancing effects of tPA. Methods—The brain striatal region of

Courtney Reddrop; Randal X. Moldrich; Philip M. Beart; Mark Farso; Gabriel T. Liberatore; David W. Howells; Karl-Uwe Petersen; Wolf-Dieter Schleuning; Robert L. Medcalf

2010-01-01

133

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

PubMed Central

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

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

2014-01-01

134

Detergent-compatible bacterial amylases.  

PubMed

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

Niyonzima, Francois N; More, Sunil S

2014-10-01

135

Enzymatic Activities in Different Strains Isolated from Healthy and Brittle Leaf Disease Affected Date Palm Leaves: Study of Amylase Production Conditions.  

PubMed

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

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

2014-11-30

136

A highly active alpha amylase from Bacillus licheniformis: directed evolution, enzyme characterization and structural analysis.  

PubMed

The stability of Bacillus licheniformis alpha-amylase (BLA) under acid condition was enhanced through direct evolution using the error-prone polymerase chain reaction. One beneficial mutation site, H281I, was obtained in BLA. The specific activity of H281I was 161/352 U/mg, which was 62.6/27.5% higher than that of the wild-type (WT) (99/276 U/mg) at pH 4.5/6.5 and 95°C. The pH optimum for H281I was decreased about 1 unit, whereas no significant changes of optimum temperature and thermostability were observed compared with the wild type (WT). The kcat/Km value of H281I was 1.7-/1.4-fold higher at pH 4.5/6.5, respectively, than that of WT. The structure model analysis indicated that the H281I mutation altered the predicted interaction between the amino acid residues at 281 and 273, thus creating a conducive local environment for substrate binding, as reflected by its decreased Km, and consequently increased the specific activity. PMID:24722373

Liu, Yihan; Fan, Shuai; Liu, Xiaoguang; Zhang, Zhimeng; Wang, Jianling; Wang, Zhengxiang; Lu, Fuping

2014-07-01

137

EFFECT OF BA AND CPPU ON PROTEASE AND ? ?-AMYLASE ACTIVITY OF IN VITRO CULTURED EXPLANTS OF ROSA HYBRIDA L  

Microsoft Academic Search

Summary. The present study was focused on the effect of N6-benzyladenine (BA) and N1-(2-chloro-4-pyridyl)-N2-phenylurea (CPPU) on protease and ?-amylase activity regarding the in vitro break and growth of lateral buds of rose (Rosa hybrida L.), cvs. Madelon and Motrea. Cv. Madelon shows strong apical growth and suppressed branching, whereas cv. Motrea is an easy branching variety. The explants were subcultured

Elena Yakimova; Veneta Kapchina-Toteva; Ivan Groshkoff; Galia Ivanov

2000-01-01

138

Adaptive significance of amylase polymorphism in Drosophila.  

E-print Network

Note Adaptive significance of amylase polymorphism in Drosophila. Analysis of the association variability at the level of the specific activity of a-amylases and their tissue-specific expression- sults indicate a homogeneous distribution of the phenotypes with a different numbers of a-amylase

Paris-Sud XI, Université de

139

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....

140

Activity of abiraterone in rechallenging two AR-expressing salivary gland adenocarcinomas, resistant to androgen-deprivation therapy.  

PubMed

Androgen-deprivation therapy (ADT) has been reported to be active in androgen receptor (AR)-expressing, relapsed/metastatic (RM), salivary gland cancers (SGCs). Abiraterone, an inhibitor of androgen synthesis, has recently been approved as a second-line treatment in hormone-resistant (HR) prostate cancer (PCa) patients. Two patients with AR-positive HR-RM adenocarcinoma, NOS of the salivary glands have been treated with abiraterone. This is the first time that this agent has been reported to be active in tumors other than HRPCa. Immunohistochemical analysis showed overexpression of EGFR, HER2, and HER3 in both untreated primary tumors. Sequencing analysis revealed a TP53 non-functional mutation in one case and a PIK3CA-activating mutation in the other. In conclusion, second line activity of ADT in AR-expressing, adenocarcinoma, NOS of salivary glands further strengthens the pathogenic and therapeutic role of AR signaling in AR-positive SGCs. PMID:24618694

Locati, Laura D; Perrone, Federica; Cortelazzi, Barbara; Imbimbo, Martina; Bossi, Paolo; Potepan, Paolo; Civelli, Enrico; Rinaldi, Gaetana; Quattrone, Pasquale; Licitra, Lisa; Pilotti, Silvana

2014-06-01

141

Identification of endo- and exo-polygalacturonase activity in Lygus hesperus (Knight) salivary glands.  

PubMed

Polygalacturonase (PG) activity found in the salivary gland apparatus of the western tarnished plant bug (WTPB, Lygus hesperus Knight) has been thought to be the main chemical cause of the damage inflicted by this mirid when feeding on its plant hosts. Early viscosity and thermal stability studies of the PG activity in L. hesperus protein extracts were difficult to interpret. Thus, it has been suggested that one or more PG protein(s) with different hydrolytic modes of action are produced by this mirid. In order to understand the quantitative complexity of the WTPB salivary PG activity, PG purification from a protein extract from salivary glands excised from L. hesperus insects was performed using affinity and ion exchange chromatography. To elucidate the qualitative complexity of the purified PGs, the digestion products generated by the PGs were separated using high performance anion exchange chromatography with pulsed amperometric detection. At least five PG proteins were detected; these differing in terms of their glycosylation, mass-to-charge ratios, and/or molecular mass. The characterization of the products generated by these PGs showed that endo- and exo-acting PGs are produced by WTPB. Although none of the PGs was purified to homogeneity, the present work provides biochemical evidence of a multiplicity of PGs that degrade the pectin component of the plant tissue in different fashions. The implications of these findings affect the understanding of WTPB feeding damage and, potentially, help identify ways to control this important crop pest. Arch. Insect Biochem. Physiol. 2008. (c) 2008 Wiley-Liss, Inc. PMID:19085947

Celorio-Mancera, Maria de la Paz; Carl Greve, L; Teuber, Larry R; Labavitch, John M

2009-02-01

142

Stability and catalytic activity of alpha-amylase from barley malt at different pressure-temperature conditions.  

PubMed

The impact of high hydrostatic pressure and temperature on the stability and catalytic activity of alpha-amylase from barley malt has been investigated. Inactivation experiments with alpha-amylase in the presence and absence of calcium ions have been carried out under combined pressure-temperature treatments in the range of 0.1-800 MPa and 30-75 degrees C. A stabilizing effect of Ca(2+) ions on the enzyme was found at all pressure-temperature combinations investigated. Kinetic analysis showed deviations of simple first-order reactions which were attributed to the presence of isoenzyme fractions. Polynomial models were used to describe the pressure-temperature dependence of the inactivation rate constants. Derived from that, pressure-temperature isokinetic diagrams were constructed, indicating synergistic and antagonistic effects of pressure and temperature on the inactivation of alpha-amylase. Pressure up to 200 MPa significantly stabilized the enzyme against temperature-induced inactivation. On the other hand, pressure also hampers the catalytic activity of alpha-amylase and a progressive deceleration of the conversion rate was detected at all temperatures investigated. However, for the overall reaction of blocked p-nitrophenyl maltoheptaoside cleavage and simultaneous occurring enzyme inactivation in ACES buffer (0.1 M, pH 5.6, 3.8 mM CaCl(2)), a maximum of substrate cleavage was identified at 152 MPa and 64 degrees C, yielding approximately 25% higher substrate conversion after 30 min, as compared to the maximum at ambient pressure and 59 degrees C. PMID:17013936

Buckow, Roman; Weiss, Ulrike; Heinz, Volker; Knorr, Dietrich

2007-05-01

143

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

PubMed Central

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

Olennikov, Daniil N.; Kashchenko, Nina I.

2014-01-01

144

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

PubMed Central

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

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

145

Periodontal status, salivary immunoglobulin, and microbial counts after short exposure to an isolated environment.  

PubMed

Salivary flow rate, immunoglobulin, and periodontal status were affected during a simulated Skylab mission. The effect is more prominent after long-duration space flights and can persist for several weeks after landing. The objective of this study was to determine the effect of a simulated Mars environment on periodontal status and levels of salivary microorganisms and immunoglobulins in the human oral cavity. Twelve healthy male volunteers were studied before, at 1 and 2 weeks, and after completion of a mission in an isolated, confined simulated Mars environment at the Mars Desert Research Station, USA. We conducted a current stress test, measured salivary immunoglobulin, cortisol, ?-amylase, salivary flow rate, and levels of plaque and salivary microbes, and assessed clinical periodontal parameters (probing depth, bleeding on probing, and clinical loss of attachment). Salivary IgG levels and Streptococcus mutans activity were significantly higher at 1 week. Values for clinical periodontal parameters (probing depth, bleeding on probing, and clinical loss of attachment) significantly differed at 1 week. Stress might be caused by the difficulty of the mission rather than the isolated environment, as mission duration was quite short. Periodontal condition might worsen due to poor oral hygiene during the mission. The present findings show that all periodontal conditions and levels of oral bacteria and stress after completion of the simulated Mars mission differed from those at baseline. To verify the relationship between stress status and periodontal health in simulated Mars missions, future studies using larger patient samples and longer follow-up will be required. PMID:23748453

Rai, Balwant; Kaur, Jasdeep

2013-01-01

146

ALPHA-AMYLASE ACTIVITY IN VARIOUS CONCENTRATIONS OF THE IONIC LIQUID, 1-BUTYL-3-METHYLIMIDAZOLIUM CHLORIDE  

Technology Transfer Automated Retrieval System (TEKTRAN)

Starch is an extremely abundant, economical and versatile industrial commodity. Many industrial uses of starch depend on hydrolyzing the polymer for the conversion of glucose and maltodextrins. Starch hydrolysis is frequently achieved by utilizing alpha-amylase, which is an endo-acting enzyme that...

147

AnAutomated, Saccharogenic Method for Determining SerumAmylase Activity  

Microsoft Academic Search

An automated adaptationof the Somogyl saccharogenic determination of serum amylase is described in which conventional AutoAnalyzer modules are used. Adequate sensitivity with short incubation is achieved by incor- porating glucose oxidase and catalase in the substrate to destroy serum glucose during incubation. Maltose and other dialyzable oligosaccharides are measured with the alkaline copper-neocuproine reaction. A simulta- neous blank run is

Wendell R. O'Neal

148

Purification of a defensin isolated from Vigna unguiculata seeds, its functional expression in Escherichia coli, and assessment of its insect alpha-amylase inhibitory activity.  

PubMed

Plant defensins make up a family of cationic antimicrobial peptides with a characteristic three-dimensional folding pattern stabilized by four disulfide bridges. The aim of this work was the purification and functional expression of a defensin from cowpea seeds and the assessment of its alpha-amylase inhibitory activity. The cDNA encoding the cowpea defensin was cloned into the pET-32 EK/LIC vector, and the resulting construct was used to transform Escherichia coli cells. The recombinant peptide was purified via affinity chromatography on a Ni Sepharose column and by reverse-phase chromatography on a C2/C18 column using HPLC. N-terminal amino acid sequencing revealed that the recombinant peptide had a similar sequence to that of the defensin isolated from seeds. The natural and recombinant defensins were submitted to the alpha-amylase inhibition assay. The cowpea seed defensin was found to inhibit alpha-amylases from the weevils Callosobruchus maculatus and Zabrotes subfasciatus. alpha-Amylase inhibition assays also showed that the recombinant defensin inhibited alpha-amylase from the weevil C. maculatus. The cowpea seed defensin and its recombinant form were unable to inhibit mammalian alpha-amylases. The three-dimensional structure of the recombinant defensin was modeled, and the resulting structure was found to be similar to those of other plant defensins. PMID:19948221

Dos Santos, Izabela S; Carvalho, André de O; de Souza-Filho, Gonçalo A; do Nascimento, Viviane V; Machado, Olga L T; Gomes, Valdirene M

2010-05-01

149

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

PubMed

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

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

150

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

PubMed Central

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

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

151

Expression of b-Amylase from Alfalfa Taproots1  

Microsoft Academic Search

Alfalfa (Medicago sativa L.) roots contain large quantities of b-amylase, but little is known about its role in vivo. We studied this by isolating a b-amylase cDNA and by examining signals that affect its expression. The b-amylase cDNA encoded a 55.95-kD polypep- tide with a deduced amino acid sequence showing high similarity to other plant b-amylases. Starch concentrations, b-amylase activities,

Joyce A. Gana; Newton E. Kalengamaliro; Suzanne M. Cunningham; Jeffrey J. Volenec

1998-01-01

152

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

PubMed Central

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

Namavar, Ferry; Sparrius, Marion; Veerman, Enno C. I.; Appelmelk, Ben J.; Vandenbroucke-Grauls, Christina M. J. E.

1998-01-01

153

Heat shock inhibits. alpha. -amylase synthesis in barley aleurone without inhibiting the activity of endoplasmic reticulum marker enzymes  

SciTech Connect

The effects of heat shock on the synthesis of {alpha}-amylase and on the membranes of the endoplasmic reticulum (ER) of barley (Hordeum vulgare) aleurone were studied. Heat shock, imposed by raising the temperature of incubation from 25{degree}C to 40{degree}C for 3 hours, inhibits the accumulation of {alpha}-amylase and other proteins in the incubation medium of barley aleurone layers treated with gibberellic acid and Ca{sup 2+}. When ER is isolated from heat-shocked aleurone layers, less newly synthesized {alpha}-amylase is found associated with this membrane system. ER membranes, as indicated by the activities of NADH cytochrome c reductase and ATP-dependent Ca{sup 2+} transport, are not destroyed by heat stress, however. Although heat shock did not reduce the activity of ER membrane marker enzymes, it altered the buoyant density of these membranes. Whereas ER from control tissue showed a peak of marker enzyme activity at 27% to 28% sucrose (1.113-1.120 grams per cubic centimeter), ER from heat-shocked tissue peaked at 30% to 32% sucrose (1.127-1.137 grams per cubic centimeter). The synthesis of a group of proteins designated as heat-shock proteins (HSPs) was stimulated by heat shock. These HSPs were localized to different compartments of the aleurone cell. Several proteins ranging from 15 to 30 kilodaltons were found in the ER and the mitochondrial/plasma membrane fractions of heat-shocked cells, but none of the HSPs accumulated in the incubation medium of heat-shocked aleurone layers.

Sticher, L.; Biswas, A.K.; Bush, D.S.; Jones, R.L. (Univ. of California, Berkeley (USA))

1990-02-01

154

Expression of ?-Amylase from Alfalfa Taproots1  

PubMed Central

Alfalfa (Medicago sativa L.) roots contain large quantities of ?-amylase, but little is known about its role in vivo. We studied this by isolating a ?-amylase cDNA and by examining signals that affect its expression. The ?-amylase cDNA encoded a 55.95-kD polypeptide with a deduced amino acid sequence showing high similarity to other plant ?-amylases. Starch concentrations, ?-amylase activities, and ?-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, ?-amylase activities, and ?-amylase transcripts were reduced significantly in roots of defoliated plants and in sucrose-deprived cell cultures. ?-Amylase transcript was high in roots of intact plants but could not be detected 2 to 8 d after defoliation. ?-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 ?-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 ?-amylase is present as a multigene family in alfalfa. Our results show no clear association between ?-amylase activity or transcript abundance and starch hydrolysis in alfalfa roots. The great abundance of ?-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

Gana, Joyce A.; Kalengamaliro, Newton E.; Cunningham, Suzanne M.; Volenec, Jeffrey J.

1998-01-01

155

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

PubMed Central

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

2012-01-01

156

Original article Amylase polymorphism  

E-print Network

Original article Amylase polymorphism in Drosophila melanogaster: haplotype frequencies in tropical December 1992) Summary - The frequencies of phenotypic haplotypes at the Amylase loci of D melanogaster by demographic bottlenecks related to recent colonizations. amylase / polymorphism / tropical populations

Boyer, Edmond

157

Inhibition of beta-amylase activity, starch degradation and sucrose formation by indole-3-acetic acid during banana ripening.  

PubMed

In order to observe the effect of indole-3-acetic acid (IAA) on carbohydrate metabolism, unripe banana (Musa acuminata AAA, cv. Nanicão) slices were infiltrated with the hormone and left to ripen under controlled conditions. The climacteric respiration burst was reduced by the action of IAA, and starch degradation and sucrose formation were delayed. Sucrose synthase (SuSy; EC 2.4.1.13) and sucrose-phosphate synthase (SPS; EC 2.4.1.14) activities and transcript levels were not affected, indicating that prevention of sucrose accumulation was not related to sucrose-metabolizing enzymes. Impairment of sucrose synthesis could be a consequence of lack of substrate, since starch degradation was inhibited. The increase in activity and transcript level of beta-amylase was delayed, indicating that this enzyme could be important in starch-to-sucrose metabolism in bananas and that it might be, at least partially, controlled at the transcriptional level. This is the first report showing that IAA can delay starch degradation, possibly affecting the activity of hydrolytic enzymes such as beta-amylase (EC 3.2.1.2). PMID:11346957

Purgatto, E; Lajolo, F M; do Nascimento, J R; Cordenunsi, B R

2001-04-01

158

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

PubMed

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

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

2014-09-01

159

Effect of tin oxide nanoparticle binding on the structure and activity of ?-amylase from Bacillus amyloliquefaciens  

NASA Astrophysics Data System (ADS)

Proteins adsorbed on nanoparticles (NPs) are being used in biotechnology, biosensors and drug delivery. However, understanding the effect of NPs on the structure of proteins is still in a nascent state. In the present paper tin oxide (SnO2) NPs were synthesized by the reaction of SnCl4·5H2O in methanol via the sol-gel method and characterized by x-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR) and transmission electron microscopy (TEM). The binding of these SnO2-NPs with ?-amylase was investigated by using UV-vis, fluorescence and circular dichroism (CD) spectroscopic techniques. A strong quenching of tryptophan fluorescence intensity in ?-amylase was observed due to formation of a ground state complex with SnO2-NPs. Far-UV CD spectra showed that the secondary structure of ?-amylase was changed in the presence of NPs. The Michaelis-Menten constant (Km), was found to be 26.96 and 28.45 mg ml - 1, while Vmax was 4.173 and 3.116 mg ml - 1 min - 1 for free and NP-bound enzyme, respectively.

Jahir Khan, Mohammad; Qayyum, Shariq; Alam, Fahad; Husain, Qayyum

2011-11-01

160

Optimizing of the formation of active BMW-amylase after in vitro refolding.  

PubMed

This study was carried out to determine the optimal folding condition of ?-amylase from Bacillus megaterium WHO using response surface methodology (RSM). A first-order model showed that three factors namely glycerol, Ca(2+) and protein concentration had the most significant effect on refolding. Analysis of the results showed that glycerol was better than the other polyols due to its effect on protein stability. Since ?-amylases are known to contain calcium ions in their structure, the presence of calcium in the refolding buffer was compulsory. The concentration of protein had the most significant quadratic effect on the response studied. A second-order polynomial model was developed to quantify the relationships between variables. It was shown that the combination of 20%(v/v) glycerol, 25 mM Ca(2+) and 0.3 (mg/ml) protein was the most efficient condition for in vitro refolding of ?-amylase. Under the optimal condition the yield of refolding was enhanced up to 7-fold. In order to analysis the size distribution in optimized and basic medium, dynamic light scattering (DLS) was fulfilled. The information gathered in this study showed that the use of solvent engineering and optimization procedure can be a general method for protein refolding. PMID:22750395

Nasrollahi, Parisa; Khajeh, Khosro; Akbari, Neda

2012-09-01

161

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

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

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

2013-05-01

162

Salivary digestive enzymes of the wheat bug, Eurygaster integriceps (Insecta: Hemiptera: Scutelleridae).  

PubMed

The digestive enzymes from salivary gland complexes (SGC) of Eurygaster integriceps, and their response to starvation and feeding were studied. Moreover, digestive amylases were partially purified and characterized by ammonium sulfate precipitation and gel filtration chromatography. The SGC are composed of two sections, the principal glands and accessory glands. The principal glands are further divided into the anterior lobes and posterior lobes. The SGC main enzyme was ?-amylase, which hydrolyzed starch better than glycogen. The other carbohydrases were also present in the SGC complexes. Enzymatic activities toward mannose (?/?-mannosidases) were little in comparison to activities against glucose (?/?-glucosidases) and galactose (?/?-galactosidases), the latter being the greatest. Acid phosphatase showed higher activity than alkaline phosphatase. There was no measurable activity for lipase and aminopeptidase. Proteolytic activity was detected against general and specific protease substrates. Activities of all enzymes were increased in response to feeding in comparison to starved insects, revealing their induction and secretion in response to feeding pulse. The SGC amylases eluted in four major peaks and post-electrophoretic detection of the ?-amylases demonstrated the existence of at least five isoamylases in the SGC. The physiological implication of these findings in pre-oral digestion of E. integriceps is discussed. PMID:24961557

Mehrabadi, Mohammad; Bandani, Ali Reza; Dastranj, Mehdi

2014-06-01

163

Molecular cloning of trypsin-like cDNAs and comparison of proteinase activities in the salivary glands and gut of the tarnished plant bug Lygus lineolaris (Heteroptera: Miridae)  

Microsoft Academic Search

Using specific proteinase inhibitors, we demonstrated that serine proteinases in the tarnished plant bug, Lyguslineolaris, 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% azocaseinase and 90% of BApNAse activities in the salivary glands were inhibited by aprotinin, benzamidine, and PMSF, whereas only

Yu Cheng Zhu; Fanrong Zeng; Brenda Oppert

2003-01-01

164

Tmem16A Encodes the Ca2+-activated Cl? Channel in Mouse Submandibular Salivary Gland Acinar Cells*  

PubMed Central

Activation of an apical Ca2+-dependent Cl? channel (CaCC) is the rate-limiting step for fluid secretion in many exocrine tissues. Here, we compared the properties of native CaCC in mouse submandibular salivary gland acinar cells to the Ca2+-gated Cl? currents generated by Tmem16A and Best2, members from two distinct families of Ca2+-activated Cl? channels found in salivary glands. Heterologous expression of Tmem16A and Best2 transcripts in HEK293 cells produced Ca2+-activated Cl? currents with time and voltage dependence and inhibitor sensitivity that resembled the Ca2+-activated Cl? current found in native salivary acinar cells. Best2?/? and Tmem16A?/? mice were used to further characterize the role of these channels in the exocrine salivary gland. The amplitude and the biophysical footprint of the Ca2+-activated Cl? current in submandibular gland acinar cells from Best2-deficient mice were the same as in wild type cells. Consistent with this observation, the fluid secretion rate in Best2 null mice was comparable with that in wild type mice. In contrast, submandibular gland acinar cells from Tmem16A?/? mice lacked a Ca2+-activated Cl? current and a Ca2+-mobilizing agonist failed to stimulate Cl? efflux, requirements for fluid secretion. Furthermore, saliva secretion was abolished by the CaCC inhibitor niflumic acid in wild type and Best2?/? mice. Our results demonstrate that both Tmem16A and Best2 generate Ca2+-activated Cl? current in vitro with similar properties to those expressed in native cells, yet only Tmem16A appears to be a critical component of the acinar Ca2+-activated Cl? channel complex that is essential for saliva production by the submandibular gland. PMID:20177062

Romanenko, Victor G.; Catalán, Marcelo A.; Brown, David A.; Putzier, Ilva; Hartzell, H. Criss; Marmorstein, Alan D.; Gonzalez-Begne, Mireya; Rock, Jason R.; Harfe, Brian D.; Melvin, James E.

2010-01-01

165

Shaddock peels (Citrus maxima) phenolic extracts inhibit ?-amylase, ?-glucosidase and angiotensin I-converting enzyme activities: a nutraceutical approach to diabetes management.  

PubMed

In this study, the interactions of free and bound phenolic-rich extracts from shaddock peels (popular in folklore for the management of diabetes and hypertension) with ?-amylase and ?-glucosidase (key enzymes linked to type-2 diabetes) and angiotensin I-converting enzyme (ACE) (key enzyme linked to hypertension) were assessed. The free phenolics of shaddock (Citrus maxima) peels were extracted with 80% acetone, while the bound phenolics were extracted from the alkaline and acid hydrolyzed residue with ethyl acetate; and their interaction with the enzymes were assessed. The phenolic extracts inhibited ?-amylase, ?-glucosidase and ACE enzyme activities in a dose-dependent manner; however, bound phenolics had significantly higher (P<0.05) ?-amylase inhibitory activities, than free phenolics, which had significantly higher (P<0.05) ACE inhibitory activities. There was no significant difference (P>0.05) in their ?-glucosidase inhibitory activities. The stronger inhibition of ?-glucosidase when compared to ?-amylase is of great pharmaceutical importance. The phenolic inhibited sodium nitroprusside induced lipid peroxidation in pancreas in a dose dependent manner. Therefore, free and bound phenolic extracts from shaddock peels could be used as nutraceutical for the management of hypertension and type-2 diabetes. PMID:22813568

Oboh, Ganiyu; Ademosun, Ayokunle O

2011-01-01

166

Potent ?-glucosidase and ?-amylase inhibitory activities of standardized 50% ethanolic extracts and sinensetin from Orthosiphon stamineus Benth as anti-diabetic mechanism  

PubMed Central

Background In the present study, we tested a 50% ethanolic extract of Orthosiphon stamineus plants and its isolated bioactive compound with respect to their ?-glucosidase and ?-amylase inhibitory activities. Methods Bioactive flavonoid sinensetin was isolated from 50% ethanolic extract of Orthosiphon stamineus. The structure of this pure compound was determined on the NMR data and the ?-glucosidase and ?-amylase inhibitory activities of isolated sinensetin and 50% ethanolic extract of Orthosiphon stamineus were evaluated. Results In vitro studies of a 50% ethanolic extract of O. stamineus and the isolated sinensetin compound showed inhibitory activity on ?-glucosidase (IC50: 4.63 and 0.66 mg/ml, respectively) and ?-amylase (IC50: 36.70 mg/ml and 1.13 mg/ml, respectively). Inhibition of these enzymes provides a strong biochemical basis for the management of type 2 diabetes via the control of glucose absorption. Conclusion Alpha-glucosidase and ?-amylase inhibition could the mechanisms through which the 50% ethanolic extract of O. stamineus and sinensetin exert their antidiabetic activity, indicating that it could have potential use in the management of non-insulin-dependent diabetes. PMID:23039079

2012-01-01

167

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

PubMed Central

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

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

2014-01-01

168

Thermal adaptation of ?-amylases: a review.  

PubMed

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

Hiteshi, Kalpana; Gupta, Reena

2014-11-01

169

Salivary alkaline phosphatase and calcium in caries-active type II diabetes mellitus patients: An in vivo study  

PubMed Central

Background: Diabetes Mellitus is a metabolic syndrome, affecting the oral health in various ways with dental caries being one of the most common problems encountered. Saliva is one of the most abundant secretions in the human body with a variety of natural protective and defence molecules bathing the oral cavity maintaining equilibrium. Its collection is easy and non-invasive. Aims: To compare and evaluate salivary alkaline phosphatase levels and calcium ion levels between caries active type II diabetes mellitus patients and non-diabetics. Materials and Methods: This study was carried out on caries-active age and gender matched 60 non-diabetic and 60 patients with known Type II diabetes mellitus subjects of age group 25-50 years with DMFT index >10. Saliva sample was collected to analyse for alkaline phosphatase enzyme and concentration of calcium ions using Agappe kits. Statistical Analysis: Student ‘t’ test was used to correlate the salivary electrolyte concentration in non- diabetic and diabetic patients with dental caries. A ‘P’ value of 0.05 or less was considered significant. Results are presented as mean ± standard deviation (X ± SD). Results: The alkaline phosphatase (ALP) activity in saliva was higher in diabetic patients when compared to that of non-diabetic patients with salivary calcium ions were significantly higher in non-diabetic individuals. Conclusion: Diabetes Mellitus patients are more prone to dental caries, hence require intervention to improve the quality of saliva. PMID:25395756

Hegde, Mithra N.; Tahiliani, Divya; Shetty, Shilpa; Devadiga, Darshana

2014-01-01

170

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

PubMed Central

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

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

2010-01-01

171

Oral digestion of a complex-carbohydrate cereal: effects of stress and relaxation on physiological and salivary measures.  

PubMed

A comprehensive study was undertaken with 12 dental hygiene students to ascertain whether the time of chewing or the degree of relaxation is more important in the oral digestion of complex carbohydrates. In addition, we studied whether the effects of stress and relaxation on salivary alpha-amylase activity was corroborated by physiologic measures. The dental hygiene students chewed an oat cereal for either 20 or 60 s while under two different orders of stress and relaxation conditions: 1) stress/20 s, stress/60 s, relax/20 s, relax/60 s; and 2) relax/20 s, relax/60 s, stress/20 s, stress/60 s. Galvanic skin resistance, pulse rate, and blood pressure (systolic and diastolic) were used to physiologically verify the effects of stress and relaxation on amylase activity. Amylase activity was judged by spectrophotometric analysis of maltose produced from a specific dilution of expectorated saliva. Results showed that the physiological measures significantly corroborated the salivary determinations of stress and relaxation and that deep relaxation was significantly more important than thorough chewing in the oral digestion of complex carbohydrates. PMID:2463752

Morse, D R; Schacterle, G R; Furst, L; Zaydenberg, M; Pollack, R L

1989-01-01

172

Original article Effect of age and exogenous amylase  

E-print Network

Original article Effect of age and exogenous amylase and protease on development of the digestive supplemented with 2 levels of enzyme preparations containing amylase and proteases up to 14 d of age. Enzyme of chymotrypsin in the pancreas and the'activity of amylase, trypsin and chymotrypsin in the intestinal contents

Paris-Sud XI, Université de

173

Salivary gland acinar cells regenerate functional glandular structures in modified hydrogels  

NASA Astrophysics Data System (ADS)

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

Pradhan, Swati

174

Diet and the evolution of human amylase gene copy number George H. Perry1,2,*, Nathaniel J. Dominy3,*, Katrina G. Claw1,4, Arthur S. Lee2, Heike  

E-print Network

Diet and the evolution of human amylase gene copy number variation George H. Perry1,2,*, Nathaniel-3. This behavioral variation raises the possibility that different selective pressures have acted on amylase, the enzyme responsible for starch hydrolysis4. We found that salivary amylase gene (AMY1) copy number

Cochran-Stafira, D. Liane

175

Daily timing of salivary cortisol responses and aerobic performance in lean and obese active females.  

PubMed

The main aim of the present study was to study the effects of morning and afternoon physical activities on cortisol responses in obese and lean women. Twenty women volunteered to participate in this study. Subjects were divided into an obese group (BMI =29.1 kg/m2) and a lean group (BMI =19 kg/m2). All subjects participated in an exercise program consisting of treadmill running at 65+/-2 % VO2max until exhaustion. In order to study effects of circadian rhythm, exercise was performed at a similar intensity and in similar environmental conditions at both 8:00 AM and 4:00 PM. Saliva specimens were collected at rest 20 minutes before activity and then immediately after the exercise in both morning and afternoon sessions. Morning and afternoon exercise resulted in a significant increase in salivary cortisol concentrations compared to basal levels in both lean and obese women (p<0.05), though the change in cortisol concentrations were higher in lean. The aerobic function of lean and obese women in the morning and afternoon showed a significant increase of 13.8 % and 5.9 %; respectively, with lean being consistently higher than obese. In conclusion, the stress response to exercise is related to circadian rhythm and individual's body weight. Based on the results of this study, it is suggested that overweight women perform exercises in the afternoon to minimize the stress response for the exercise volume performed (Tab. 1, Fig. 3, Ref. 39). Full Text in free PDF www.bmj.sk. PMID:21585131

Azarbayjani, M A; Vaezepor, F; Rasaee, M J; Tojaril, F; Pournemati, P; Jourkesh, M; Ostojic, S M; Stannard, S R

2011-01-01

176

Immobilization of Trichoderma harzianum ?-amylase on treated wool: optimization and characterization.  

PubMed

?-Amylase from Trichoderma harzianum was covalently immobilized on activated wool by cyanuric chloride. Immobilized ?-amylase exhibited 75% of its initial activity after 10 runs. The soluble and immobilized ?-amylases exhibited maximum activity at pH values 6.0 and 6.5, respectively. The immobilized enzyme was more thermally stable than the soluble one. Various substrates were hydrolyzed by immobilized ?-amylase with high efficiencies compared to those of soluble ?-amylase. The inhibition of the immobilized ?-amylase by metal ions was low as compared with soluble enzyme. On the basis of the results obtained, immobilized ?-amylase could be employed in the saccharification of starch processing. PMID:24932573

Mohamed, Saleh A; Khan, Jalaluddin A; Al-Bar, Omar A M; El-Shishtawy, Reda M

2014-01-01

177

Factors affecting antimicrobial activity of MUC7 12-mer, a human salivary mucin-derived peptide  

PubMed Central

Background MUC7 12-mer (RKSYKCLHKRCR), a cationic antimicrobial peptide derived from the human low-molecular-weight salivary mucin MUC7, possesses potent antimicrobial activity in vitro. In order to evaluate the potential therapeutic application of the MUC7 12-mer, we examined the effects of mono- and divalent cations, EDTA, pH, and temperature on its antimicrobial activity. Methods Minimal Inhibitory Concentrations (MICs) were determined using a liquid growth inhibition assay in 96-well microtiter plates. MUC7 12-mer was added at concentrations of 1.56–50 ?M. MICs were determined at three endpoints: MIC-0, MIC-1, and MIC-2 (the lowest drug concentration showing 10%, 25% and 50% of growth, respectively). To examine the effect of salts or EDTA, a checkerboard microdilution technique was used. Fractional inhibitory concentration index (FICi) was calculated on the basis of MIC-0. The viability of microbial cells treated with MUC7 12-mer in the presence of sodium or potassium was also determined by killing assay or flow cytometry. Results The MICs of MUC7 12-mer against organisms tested ranged from 6.25–50 ?M. For C. albicans, antagonism (FICi 4.5) was observed for the combination of MUC7 12-mer and calcium; however, there was synergism (FICi 0.22) between MUC7 12-mer and EDTA, and the synergism was retained in the presence of calcium at its physiological concentration (1–2 mM). No antagonism but additivity or indifference (FICi 0.55–2.5) was observed for the combination of MUC7 12-mer and each K+, Na+, Mg2+, or Zn2+. MUC7 12-mer peptide (at 25 ?M) also exerted killing activity in the presence of NaCl, (up to 25 mM for C. albicans and up to 150 mM for E. coli, a physiological concentration of sodium in the oral cavity and serum, respectively) and retained candidacidal activity in the presence of KCl (up to 40 mM). The peptide exhibited higher inhibitory activity against C. albicans at pH 7, 8, and 9 than at pH 5 and 6, and temperature up to 60°C did not affect the activity. Conclusion MUC7 12-mer peptide is effective anticandidal agent at physiological concentrations of variety of ions in the oral cavity. These results suggest that, especially in combination with EDTA, it could potentially be applied as an alternative therapeutic agent for the treatment of human oral candidiasis. PMID:17996119

Wei, Guo-Xian; Campagna, Alexander N; Bobek, Libuse A

2007-01-01

178

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

PubMed Central

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

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

2014-01-01

179

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

ERIC Educational Resources Information Center

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…

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

2008-01-01

180

THE SALIVARY CATECHOL OXIDASE\\/PEROXIDASE ACTIVITIES OF THE MOSQUITO ANOPHELES ALBIMANUS  

Microsoft Academic Search

Summary Salivary gland homogenates from adult female Anopheles albimanus mosquitoes relaxed aortic rings preconstricted with noradrenaline (NA). This relaxation is slow and is due to destruction of NA. Incubation of NA with the homogenate yielded a product with a spectrum consistent with the corresponding adrenochrome. Oxidation of NA was enhanced by a superoxide generation system and inhibited by the combined

JOSÉ M. C. RIBEIRO; ROBERTO H. NUSSENZVEIG

1993-01-01

181

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

PubMed

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

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

2013-01-01

182

Molecular identification of four different alpha-amylase inhibitors from baru (Dipteryx alata) seeds with activity toward insect enzymes.  

PubMed

The endophytic bruchid pest Callosobruchus maculatus causes severe damage to storage cowpea seeds, leading to economical losses. For this reason the use of alpha-amylase inhibitors to interfere with the pest digestion process has been an interesting alternative to control bruchids. With this aim, alpha-amylase inhibitors from baru seeds (Dipteryx alata) were isolated by affinity chromatographic procedures, causing enhanced inhibition of C. maculatus and Anthonomus grandis alpha-amylases. To attempt further purification, this fraction was applied onto a reversed-phase HPLC column, generating four peaks with remarkable inhibition toward C. maculatus alpha-amylases. SDS-PAGE and MALDI-ToF analysis identified major proteins of approximately 5.0, 11.0, 20.0 and 55 kDa that showed alpha-amylase inhibition. Results of in vivo bioassays using artificial seeds containing 1.0% (w/w) of baru crude extract revealed 40% cowpea weevil larvae mortality. These results provide evidence that several alpha-amylase inhibitors classes, with biotechnological potential, can be isolated from a single plant species. PMID:17669264

Bonavides, Krishna B; Pelegrini, Patrícia B; Laumann, Raúl A; Grossi-de-Sá, Maria F; Bloch, Carlos; Melo, Jorge A T; Quirino, Betania F; Noronha, Eliane F; Franco, Octávio L

2007-07-31

183

Salivary Surrogates of Plasma Nitrite and Catecholamines during a 21-Week Training Season in Swimmers  

PubMed Central

The collection of samples of saliva is noninvasive and straightforward, which turns saliva into an ideal fluid for monitoring the adaptive response to training. Here, we investigated the response of the salivary proteins alpha-amylase (sAA), chromogranin A (sCgA), and the concentration of total protein (sTP) as well as salivary nitrite (sNO2) in relation to plasma catecholamines and plasma nitrite (pNO2), respectively. The variation in these markers was compared to the intensity and load of training during a 21-week training season in 12 elite swimmers. Overall, the salivary proteins tracked the concentration of plasma adrenaline and were inversely correlated with the training outcomes. No correlations were observed between sNO2 and pNO2. However, sNO2 correlated positively with the intensity and load of training. We argue that the decrease in sympathetic activity is responsible for the decrease in the concentration of proteins throughout the training season. Furthermore, the increase in nitrite is likely to reflect changes in hemodynamics and regulation of vascular tone. The association of the salivary markers with the training outcomes underlines their potential as noninvasive markers of training status in professional athletes. PMID:23700456

Díaz Gómez, Miguel Mauricio; Bocanegra Jaramillo, Olga Lucia; Teixeira, Renata Roland; Espindola, Foued Salmen

2013-01-01

184

Covalent immobilization of ?-amylase onto poly(2-hydroxyethyl methacrylate) and poly(styrene -2-hydroxyethyl methacrylate) microspheres and the effect of Ca 2+ ions on the enzyme activity  

Microsoft Academic Search

?-Amylase (1,4-?-d-glucan-glucanohydrolase; EC 3.2.1.1, Type VI-B from porcine pancreas, extra pure 29 units mg?1) was covalently immobilized on poly (2-hydroxyethyl methacrylate), p(HEMA), and poly (styrene- 2- hydroxyethyl methacrylate), p(St-HEMA) microspheres, which were activated by using epichlorohydrin (ECH). The properties of the immobilized enzyme were investigated and compared with those of the free enzyme. For the assays carried out at 25°C

Hayrettin Tümtürk; Serpil Aksoy; Nesrin Has?rc?

2000-01-01

185

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

PubMed Central

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

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

186

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

PubMed Central

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

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

187

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

PubMed

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

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

2015-02-01

188

Expression of beta-amylase from alfalfa taproots.  

PubMed

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

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

1998-12-01

189

Is the difference in alpha-amylase activity in the strains of Drosophila melanogaster with different allozymes due to transcriptional or posttranscriptional control?  

PubMed

The alpha-amylase in Drosophila melanogaster is a highly polymorphic enzyme, at both the allozyme level and the specific activity level. This enzyme changes its specific activity drastically depending on both food conditions and developmental stages, and it has been suggested that the ability to change its activity depending on the source of food has positive correlation with fitness. But the cause of the difference of specific activity among strains and food compositions is not known. In order to investigate the cause of these differences, we measured both the specific activity of amylase and the relative amount of Amy mRNA in eight strains of D. melanogaster with different electromorphs, in two food environments and two developmental stages. We found the following. First, the food-dependent activity change is regulated at the transcription level. Second, there was a significant correlation between specific activity and mRNA level among strains. So 80 to 90% of the specific activity difference can be explained by differences in the level of mRNAs, but the remaining part cannot. Finally, there were significant differences in specific activity per mRNA both among strains and between developmental stages. This suggests that there are differences in the catalytic efficiency of each allozyme, strain- or stage-specific translation rate, enzyme stability, or differential use of two Amy loci. PMID:10690430

Yamate, N; Yamazaki, T

1999-12-01

190

Radionuclide salivary gland imaging  

SciTech Connect

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.

Mishkin, F.S.

1981-10-01

191

Water Stress Enhances Expression of an ?-Amylase Gene in Barley Leaves  

PubMed Central

The amylases of the second leaves of barley seedlings (Hordeum vulgare L. cv Betzes) were resolved into eight isozymes by isoelectric focusing, seven of which were ?-amylase and the other, ?-amylase. The ?-amylase had the same isoelectric point as one of the gibberellin-induced ?-amylase isozymes in the aleurone layer. This and other enzyme characteristics indicated that the leaf isozyme corresponded to the type A aleurone ?-amylase (low pI group). Crossing experiments indicated that leaf and type A aleurone isozymes resulted from expression of the same genes. In unwatered seedlings, leaf ?-amylase increased as leaf water potential decreased and ABA increased. Water stress had no effect on ?-amylase. ?-Amylase occurred uniformly along the length of the leaf but ?-amylase was concentrated in the basal half of the leaf. Cell fractionation studies indicated that none of the leaf ?-amylase occurred inside chloroplasts. Leaf radiolabeling experiments followed by extraction of ?-amylase by affinity chromatography and immunoprecipitation showed that increase of ?-amylase activity involved synthesis of the enzyme. However, water stress caused no major change in total protein synthesis. Hybridization of a radiolabeled ?-amylase-related cDNA clone to size fractionated RNA showed that water-stressed leaves contained much more ?-amylase mRNA than unstressed plants. The results of these and other studies indicate that regulation of gene expression may be a component in water-stress induced metabolic changes. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 6 Fig. 7 Fig. 8 Fig. 9 PMID:16664625

Jacobsen, John V.; Hanson, Andrew D.; Chandler, Peter C.

1986-01-01

192

Immunohistochemical expression of the oncogenic molecules active Stat3 and survivin in benign and malignant salivary gland tumors  

PubMed Central

Objective Signal transducer and activator of transcription 3 (Stat3) and survivin have been shown to exert oncogenic effects in various human neoplasms. The purpose of this study was to evaluate the expression of the tyrosine phosphorylated (active) Stat3 and survivin in various benign and malignant salivary gland tumors (SGTs). Study design Eighty-six SGTs (65 malignant and 21 benign tumors of various histopathologic subtypes) were immunohistochemically stained with anti-survivin or anti-phosphorylated tyrosine-705 (p-tyr) Stat3 antibodies. Immunohistochemical reactivity was graded in a semi-quantitative manner; a combined score of immunohistochemical positivity (0–6) was calculated for each tumor by adding the individual scores for percentage of tumor cells (0–3) and intensity of staining (0–3). Results Survivin was immunohistochemically detected in all studied benign and malignant SGTs; p-tyr Stat3 was also detected in the majority (91%) of SGTs. The average combined scores for survivin and p-tyr Stat3 immunohistochemical expression in the studied malignant SGTs was 4.40 and 3.35, respectively; the corresponding combined scores for survivin and p-tyr Stat3 in the studied benign SGTs were 4.37 and 3.22, respectively. No statistically significant differences (p>0.05) in p-tyr Stat3 or survivin expression were detected between the benign and malignant groups, or among the various examined histopathological subtypes of SGTs. In contrast, normal salivary gland elements in the vicinity of the studied tumors revealed only weak and focal survivin or p-tyr Stat3 immunoreactivity, mainly localized to ductal and mucous cells. Conclusions Our data indicate an almost universal expression of activated Stat3 and survivin in benign and malignant SGTs. Considering the well-established proliferative and anti-apoptotic properties of these molecules and their functional interrelationship, selective targeting techniques against Stat3 and/or survivin may represent promising therapeutic strategies against neoplasms of salivary gland origin. PMID:19272817

Nikitakis, Nikolaos G.; Scheper, Mark A.; Papanicolaou, Vasileios S.; Sklavounou, Alexandra; Sauk, John J.

2009-01-01

193

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

PubMed Central

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

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

2010-01-01

194

Effect of betulin-containing extract from birch tree bark on ?-amylase activity in vitro and on weight gain of broiler chickens in vivo.  

PubMed

In vitro effect of betulin-containing extract from Betula pendula Roth. bark on alpha-amylase activity was studied, the kinetic mechanism of interaction was proposed and in vivo effect of betulin-containing extract on weight gain and meat quality of broiler chickens was evaluated. The highest level of inhibitory activity (20%) was detected in extract concentration of 1,000 mg/L. Increased extract concentration did not lead to increased enzyme inhibition. Using Dixon and Cornish-Bowden coordinates, the competitive mechanism of inhibition was demonstrated. Calculated kinetic parameters were: Km equal to 0.6 mg/mL, Vmax equal to 2.6 and 2.1 mM/min from Lineweaver-Burk and Dixon coordinates, respectively and Ki equal to 3,670?±?230 mg/mL. The partial inhibition of enzyme indicates the existence of low concentration of active inhibitory form, which reaches saturation level with increased extract concentration in applied suspension. Therefore, Ki has an apparent constant character. This partial inhibition of amylase activity observed in in vitro assay did not affect weight gain and meat quality of broiler chickens during in vivo assay. Rather, the tendency to increase the weight of edible parts and muscles compared to diet without additive suggests that the extract may be a potential food additive in poultry farming. Additionally, it could be a source for further pharmaceutical and pharmacological research. PMID:24445672

Ilyina, Anna; Arredondo-Valdés, Roberto; Farkhutdinov, Salavat; Segura-Ceniceros, Elda Patricia; Martínez-Hernández, José Luis; Zaynullin, Radik; Kunakova, Rayhana

2014-03-01

195

Mode of alpha-amylase production by the shochu koji mold Aspergillus kawachii.  

PubMed

Aspergillus kawachii produces two kinds of alpha-amylase, one is an acid-unstable alpha-amylase and the other is an acid-stable alpha-amylase. Because the quality of the shochu depends strongly on the activities of the alpha-amylases, the culture conditions under which these alpha-amylases are produced were examined. In liquid culture, acid-unstable alpha-amylase was produced abundantly, but, acid-stable alpha-amylase was not produced. The acid-unstable alpha-amylase was produced significantly when glycerol or glucose was used as a carbon source, similarly to the use of inducers such as starch or maltose. In liquid culture, A. kawachii assimilated starch at pH 3.0, but no alpha-amylase activity was recognized in the medium. Instead, the alpha-amylase was found to be trapped in the cell wall. The trapped form was identified as acid-unstable alpha-amylase. Usually, acid-unstable alpha-amylase is unstable at pH 3.0, so its stability appeared to be due to its immobilization in the cell wall. In solid-state culture, both kinds of alpha-amylase were produced. The production of acid-stable alpha-amylase seems to be solid-state culture-specific and was affected by the moisture content in the solid medium. PMID:14586108

Nagamine, Kazuki; Murashima, Kenji; Kato, Taku; Shimoi, Hitoshi; Ito, Kiyoshi

2003-10-01

196

Salivary gland and peripheral blood T helper 1 and 2 cell activity in Sjo?gren’s syndrome compared with non-Sjo?gren’s sicca syndrome  

Microsoft Academic Search

Objectives: To investigate whether differences in T helper (Th) 1 and Th2 cell activity in salivary glands (“local”) or (“peripheral”) blood can discriminate between Sjo?gren’s syndrome (SS) and non-Sjo?gren’s sicca syndrome (nSS-sicca). Additionally, to study relationships of local and peripheral Th cell activities with each other and with disease activity measures.Methods: 62 sicca patients (32 with SS, 30 with nSS-sicca)

J M van Woerkom; A A Kruize; M J G Wenting-van Wijk; E Knol; I C Bihari; J W G Jacobs; J W J Bijlsma; F P J G Lafeber; J A G van Roon

2005-01-01

197

Effects of temperature on the endogenous activity and synaptic interactions of the salivary burster neurones in the terrestrial slug Limax maximus.  

PubMed

(1) The activity of the endogenously active salivary burster neurones (SBs) shows temperature acclimation and has characteristic cold and warm blockade temperatures. (2) Temperature acclimation affects the upper and lower limits of the temperature range over which SBs are active. The absolute range, in centigrade degrees, during warming, is unaffected by acclimation. (3) Acclimatization of burster activity is a slow response to the mean ambient temperature. (4) There is increased synchrony of activity between the right and left salivary bursters at low temperature which is correlated with an increased electrical coupling between the SBs and protractor motoneurones (B7s). There is a corresponding increase in the input resistance of B7 at low temperatures. PMID:7108437

Prior, D J; Grega, D S

1982-06-01

198

Effect of introducing a disulphide bond between the A and C domains on the activity and stability of Saccharomycopsis fibuligera R64 ?-amylase.  

PubMed

Native enzyme and a mutant containing an extra disulphide bridge of recombinant Saccharomycopsis fibuligera R64 ?-amylase, designated as Sfamy01 and Sfamy02, respectively, have successfully been overexpressed in the yeast Pichia pastoris KM71H. The purified ?-amylase variants demonstrated starch hydrolysis resulting in a mixture of maltose, maltotriose, and glucose, similar to the wild type enzyme. Introduction of the disulphide bridge shifted the melting temperature (TM) from 54.5 to 56°C and nearly tripled the enzyme half-life time at 65°C. The two variants have similar kcat/KM values. Similarly, inhibition by acarbose was only slightly affected, with the IC50 of Sfamy02 for acarbose being 40±3.4?M, while that of Sfamy01 was 31±3.9?M. On the other hand, the IC50 of Sfamy02 for EDTA was 0.45mM, nearly two times lower than that of Sfamy01 at 0.77mM. These results show that the introduction of a disulphide bridge had little effect on the enzyme activity, but made the enzyme more susceptible to calcium ion extraction. Altogether, the new disulphide bridge improved the enzyme stability without affecting its activity, although minor changes in the active site environment cannot be excluded. PMID:25533400

Natalia, Dessy; Vidilaseris, Keni; Ismaya, Wangsa T; Puspasari, Fernita; Prawira, Iman; Hasan, Khomaini; Fibriansah, Guntur; Permentier, Hjalmar P; Nurachman, Zeily; Subroto, Toto; Dijkstra, Bauke W; Soemitro, Soetijoso

2015-02-10

199

Effect of salivary gland adenocarcinoma cell-derived alpha-N-acetylgalactosaminidase on the bioactivity of macrophage activating factor.  

PubMed

The aim of this study was to clarify the effects of alpha-N-acetylgalactosaminidase (alpha-NaGalase) produced by human salivary gland adenocarcinoma (SGA) cells on the bioactivity of macrophage-activating factor (GcMAF). High exo-alpha-NaGalase activity was detected in the SGA cell line HSG. HSG alpha-NaGalase had both exo- and endo-enzyme activities, cleaving the Gal-GalNAc and GalNAc residues linked to Thr/Ser but not releasing the [NeuAc2-6]GalNac residue. Furthermore, GcMAF enzymatically prepared from the Gc protein enhanced the superoxide-generation capacity and phagocytic activity of monocytes/macrophages. However, GcMAF treated with purified alpha-NaGalase did not exhibit these effects. Thus, HSG possesses the capacity to produce larger quantities of alpha-NaGalase, which inactivates GcMAF produced from Gc protein, resulting in reduced phagocytic activity and superoxide-generation capacity of monocytes/macrophages. The present data strongly suggest that HSG alpha-NaGalase acts as an immunodeficiency factor in cancer patients. PMID:14767536

Matsuura, Takashi; Uematsu, Takashi; Yamaoka, Minoru; Furusawa, Kiyofumi

2004-03-01

200

Purification and characterisation of a malto-oligosaccharide-forming amylase active at high pH from Bacillus clausii BT-21.  

PubMed

Bacillus clausii BT-21 produced an extracellular malto-oligosaccharide-forming amylase active at high pH when grown on starch substrates. The enzyme was purified to homogeneity by affinity and anion-exchange chromatography. The molecular weight of the enzyme estimated by sodium dodecyl sulfate polyacrylamide electrophoresis was 101 kDa. The enzyme showed an optimum of activity at pH 9.5 and 55 degrees C. Maltohexaose was detected as the main initially formed starch hydrolysis product. Maltotetraose and maltose were the main products obtained after hydrolysis of starch by the enzyme for an extended period of time and were not further degraded. The enzyme readily hydrolysed soluble starch, amylopectin and amylose, while cyclodextrins, pullulan or dextran were not degraded. The mode of action during hydrolysis of starch indicated an exo-acting type of amylolytic enzyme mainly producing maltohexaose and maltotetraose. Amino acid sequencing of the enzyme revealed high homology with the maltohexaose-forming amylase from Bacillus sp. H-167. PMID:11086690

Duedahl-Olesen, L; Kragh, K M; Zimmermann, W

2000-10-20

201

Amylase and cyclic amp receptor protein expression in human diabetic parotid glands  

E-print Network

Amylase and cyclic amp receptor protein expression in human diabetic parotid glands Monica Piras1 BACKGROUND: Salivary dysfunction and oral disorders have been described in both type 1 and type 2 diabetes mellitus. However, the cellular and molecular conse- quences of diabetes on oral tissues remain to be ascer

Terasaki, Mark

202

Suppression of white light generation (supercontinuum) in biological media: a pilot study using human salivary proteins  

NASA Astrophysics Data System (ADS)

Propagation of ultrashort pulses of intense, infrared light through transparent medium gives rise to a visually spectacular phenomenon known as supercontinuum (white light) generation wherein the spectrum of transmitted light is very considerably broader than that of the incident light. We have studied the propagation of ultrafast (<45 fs) pulses of intense infrared light through biological media (water, and water doped with salivary proteins) which reveal that white light generation is severely suppressed in the presence of a major salivary protein, ?-amylase.

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

2007-02-01

203

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

PubMed Central

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

Singh, Sanamdeep; Bali, Vrinda; Mangla, Jyoti

2014-01-01

204

Induction of a Peptide with Activity against a Broad Spectrum of Pathogens in the Aedes aegypti Salivary Gland, following Infection with Dengue Virus  

PubMed Central

The ultimate stage of the transmission of Dengue Virus (DENV) to man is strongly dependent on crosstalk between the virus and the immune system of its vector Aedes aegypti (Ae. aegypti). Infection of the mosquito's salivary glands by DENV is the final step prior to viral transmission. Therefore, in the present study, we have determined the modulatory effects of DENV infection on the immune response in this organ by carrying out a functional genomic analysis of uninfected salivary glands and salivary glands of female Ae. aegypti mosquitoes infected with DENV. We have shown that DENV infection of salivary glands strongly up-regulates the expression of genes that encode proteins involved in the vector's innate immune response, including the immune deficiency (IMD) and Toll signalling pathways, and that it induces the expression of the gene encoding a putative anti-bacterial, cecropin-like, peptide (AAEL000598). Both the chemically synthesized non-cleaved, signal peptide-containing gene product of AAEL000598, and the cleaved, mature form, were found to exert, in addition to antibacterial activity, anti-DENV and anti-Chikungunya viral activity. However, in contrast to the mature form, the immature cecropin peptide was far more effective against Chikungunya virus (CHIKV) and, furthermore, had strong anti-parasite activity as shown by its ability to kill Leishmania spp. Results from circular dichroism analysis showed that the immature form more readily adopts a helical conformation which would help it to cause membrane permeabilization, thus permitting its transfer across hydrophobic cell surfaces, which may explain the difference in the anti-pathogenic activity between the two forms. The present study underscores not only the importance of DENV-induced cecropin in the innate immune response of Ae. aegypti, but also emphasizes the broad-spectrum anti-pathogenic activity of the immature, signal peptide-containing form of this peptide. PMID:21249175

Patramool, Sirilaksana; Dumas, Emilie; Wasinpiyamongkol, Ladawan; Saune, Laure; Hamel, Rodolphe; Bernard, Eric; Sereno, Denis; Thomas, Frédéric; Piquemal, David; Yssel, Hans; Briant, Laurence; Missé, Dorothée

2011-01-01

205

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

PubMed Central

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

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

2014-01-01

206

Organophosphate pesticide environmental exposure: analysis of salivary cholinesterase and carboxilesterase activities in preschool children and their mothers.  

PubMed

A pilot study was conducted to evaluate the usefulness of salivary cholinesterase and carboxylesterase as biomarkers of exposure to environmental organophosphate pesticides. Ninety samples were obtained from women and 62 samples from their preschool-aged children who live near an agricultural area of the Upper Valley of the Negro River (Patagonia, Argentina) where pesticides are applied 6 months a year. Each participant donated two samples under similar conditions: one in the pre-exposure period and another during the pulverization period. Demographic information, potential confounders, and risk behaviors were registered. Active or passive smoking had no effect on these enzyme activities in either group. During the pulverization period, cholinesterase activity was not detectable in 76% of the children's samples and 23% of the mothers' samples. Comparing samples collected during the pulverization period with respect to the pre-pulverization period, the average mother and child cholinesterase activity decreased by 65.7% (p?activity decreased by 27.5% (p?activity in the pulverization period was associated to the habit of eating dust outdoors (p?activity were between 70-100% and 0-29%, respectively, in both groups studied. This shows that at the same level of exposure, cholinesterase was more sensitive to inhibition than carboxylesterase. Therefore, carboxylesterase might more properly reflect the degree of environmental organophosphate exposure and may have potential as a novel tool for biomonitoring. PMID:21739279

Bulgaroni, Vanina; Rovedatti, María Gabriela; Sabino, Guillermo; Magnarelli, Gladis

2012-05-01

207

Purification and characterization of ?-Amylase from Miswak Salvadora persica  

PubMed Central

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

2014-01-01

208

Immobilization of ?-Amylase onto Luffa operculata Fibers  

PubMed Central

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

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

2013-01-01

209

Salivary gland infections  

MedlinePLUS

... Physiology of the salivary glands. In: Cummings CW, Flint PW, Haughey BH, et al, eds. Otolaryngology: Head & ... disorders of the salivary glands. In: Cummings CW, Flint PW, Haughey BH, et al, eds. Otolaryngology: Head & ...

210

Alpha-amylase inhibitor, CS-1036 binds to serum amylase in a concentration-dependent and saturable manner.  

PubMed

(2R,3R,4R)-4-hydroxy-2-(hydroxymethyl)pyrrolidin-3-yl 4-O-(6-deoxy-?-D-glucopyranosyl)-?-D-glucopyranoside (CS-1036), which is an ?-amylase inhibitor, exhibited biphasic and sustained elimination with a long t1/2 (18.4-30.0 hours) in rats and monkeys, but exhibited a short t1/2 (3.7-7.9 hours) in humans. To clarify the species differences in the t1/2, the plasma protein binding of CS-1036 was evaluated by ultrafiltration. A concentration-dependent and saturable plasma protein binding of CS-1036 was observed in rats and monkeys with the dissociation rate constant (KD) of 8.95 and 27.2 nM, and maximal binding capacity (Bmax) of 52.8 and 22.1 nM, respectively. By the assessments of the recombinant amylase and immunoprecipitation, the major binding protein of CS-1036 in rats was identified as salivary amylase (KD 5.64 nM). CS-1036 also showed concentration-dependent and saturable binding to human salivary and pancreatic amylase, with similar binding affinity in rats. However, the protein binding of CS-1036 was constant in human plasma (?10.2%) due to the lower serum amylase level compared with rats and monkeys. From the calculation of the unbound fraction (fu) in plasma based on in vitro KD and Bmax, the dose-dependent increase in fu after oral administration is speculated to lead to a dose-dependent increase in total body clearance and a high area under the curve/dose at lower doses, such as 0.3 mg/kg in rats. PMID:24319124

Honda, Tomohiro; Kaneno-Urasaki, Yoko; Ito, Takashi; Kimura, Takako; Matsushima, Nobuko; Okabe, Hiromi; Yamasaki, Atsushi; Izumi, Takashi

2014-03-01

211

Differential modulation of voltage-activated conductances by intracellular and extracellular cyclic nucleotides in leech salivary glands.  

PubMed Central

1. Two-electrode voltage clamp was used to study the effects of adenosine 3':5'-cyclic monophosphate (cyclic AMP) and guanosine 3':5'-cyclic monophosphate (cyclic GMP) on voltage-dependent ion channels in salivary gland cells of the leech, Haementeria ghilianii. 2. Intracellular cyclic AMP specifically blocked delayed rectifier K+ channels. This was shown by use of 3-isobutyl-1-methylxanthine (IBMX, a phosphodiesterase inhibitor), forskolin (an activator of adenylyl cyclase) and intracellular injection of cyclic AMP and its dibutyryl and 8-bromo analogues. Cyclic AMP appeared to be the second messenger for the putative neuroglandular transmitter, 5-hydroxytryptamine. 3. Intracellular injection of cyclic GMP specifically potentiated high-voltage-activated (HVA) Ca2+ current and the effect was mimicked by zaprinast, an inhibitor of cyclic GMP-dependent phosphodiesterase. 4. Extracellularly, cyclic GMP and cyclic AMP specifically decreased the amplitude and increased the rate of inactivation of HVA Ca2+ current. These effects of the cyclic nucleotides are identical to those known for extracellular ATP, which activates a presumed purinoceptor. The pyrimidine nucleotide, UTP, was almost equipotent to ATP (threshold dose < 10(-6) M), indicative of a vertebrate-type nucleotide receptor. However, suramin (5 x 10(-5) M), a non-specific P2-receptor antagonist, failed to block the effects of 5 x 10(-6) M ATP (higher suramin doses could not be reliably tested because of the depolarization and increase in membrane conductance produced by the drug). 5. Activation of the putative purinoceptor by ATP did not affect inward rectifier Na+/K+ current which is known to be potentiated by intracellular cyclic AMP and reduced by intracellular cyclic GMP. 6. The preparation may provide a useful model for study of nucleotide actions, and interactions, in channel modulation. It has technical advantages such as large cells (1200 microns in diameter) which lack intercellular coupling and may be individually dissected for biochemical studies. PMID:8528570

Everill, B.; Berry, M. S.

1995-01-01

212

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

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

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

2014-01-01

213

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

PubMed Central

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

Maddi, A; Haase, EM; Scannapieco, FA

2014-01-01

214

A 28-fold increase in secretory protein synthesis is associated with DNA puff activity in the salivary gland of Bradysia hygida (Diptera, Sciaridae).  

PubMed

When the first group of DNA puffs is active in the salivary gland regions S1 and S3 of Bradysia hygida larvae, there is a large increase in the production and secretion of new salivary proteins demonstrable by [3H]-Leu incorporation. The present study shows that protein separation by SDS-PAGE and detection by fluorography demonstrated that these polypeptides range in molecular mass from about 23 to 100 kDa. Furthermore, these proteins were synthesized mainly in the S1 and S3 salivary gland regions where the DNA puffs C7, C5, C4 and B10 are conspicuous, while in the S2 region protein synthesis was very low. Others have shown that the extent of amplification for DNA sequences that code for mRNA in the DNA puffs C4 and B10 was about 22 and 10 times, respectively. The present data for this group of DNA puffs are consistent with the proposition that gene amplification is necessary to provide some cells with additional gene copies for the production of massive amounts of proteins within a short period of time. PMID:9283627

de-Almeida, J C

1997-05-01

215

Rescue of Salivary Gland Function after Stem Cell Transplantation in Irradiated Glands  

PubMed Central

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

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

216

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

ERIC Educational Resources Information Center

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)

Freeland, P. W.

1972-01-01

217

What interactions drive the salivary mucosal pellicle formation?  

PubMed Central

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

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

2014-01-01

218

What interactions drive the salivary mucosal pellicle formation?  

PubMed

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

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

2014-08-01

219

Salivary proteins of plant-feeding hemipteroids - implication in phytophagy.  

PubMed

Many hemipteroids are major pests and vectors of microbial pathogens, infecting crops. Saliva of the hemipteroids is critical in enabling them to be voracious feeders on plants, including the economically important ones. A plethora of hemipteroid salivary enzymes is known to inflict stress in plants, either by degrading the plant tissue or by affecting their normal metabolism. Hemipteroids utilize one of the following three strategies of feeding behaviour: salivary sheath feeding, osmotic-pump feeding and cell-rupture feeding. The last strategy also includes several different tactics such as lacerate-and-flush, lacerate-and-sip and macerate-and-flush. Understanding hemipteroid feeding mechanisms is critical, since feeding behaviour directs salivary composition. Saliva of the Heteroptera that are specialized as fruit and seed feeders, includes cell-degrading enzymes, auchenorrhynchan salivary composition also predominantly consists of cell-degrading enzymes such as amylase and protease, whereas that of the Sternorhyncha includes a variety of allelochemical-detoxifying enzymes. Little is known about the salivary composition of the Thysanoptera. Cell-degrading proteins such as amylase, pectinase, cellulase and pectinesterase enable stylet entry into the plant tissue. In contrast, enzymes such as glutathione peroxidase, laccase and trehalase detoxify plant chemicals, enabling the circumvention of plant-defence mechanisms. Salivary enzymes such as M1-zinc metalloprotease and CLIP-domain serine protease as in Acyrthosiphon pisum (Aphididae), and non-enzymatic proteins such as apolipophorin, ficolin-3-like protein and 'lava-lamp' protein as in Diuraphis noxia (Aphididae) have the capacity to alter host-plant-defence mechanisms. A majority of the hemipteroids feed on phloem, hence Ca++-binding proteins such as C002 protein, calreticulin-like isoform 1 and calmodulin (critical for preventing sieve-plate occlusion) are increasingly being recognized in hemipteroid-plant interactions. Determination of a staggering variety of proteins shows the complexity of hemipteroid saliva: effector proteins localized in hemipteran saliva suggest a similarity to the physiology of pathogen-plant interactions. PMID:24280006

Sharma, A; Khan, A N; Subrahmanyam, S; Raman, A; Taylor, G S; Fletcher, M J

2014-04-01

220

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

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

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

2013-01-01

221

Production of ?-Amylase by the Ruminal Anaerobic Fungus Neocallimastix frontalis  

PubMed Central

?-Amylase production was examined in the ruminal anaerobic fungus Neocallimastix frontalis. The enzyme was released mainly into the culture fluid and had temperature and pH optima of 55°C and 5.5, respectively, and the apparent Km for starch was 0.8 mg ml?1. The products of ?-amylase action were mainly maltotriose, maltotetraose, and longer-chain oligosaccharides. No activity of the enzyme was observed towards these compounds or pullulan, but activity on amylose was similar to starch. Evidence for the endo action of ?-amylase was also obtained from experiments which showed that the reduction in iodine-staining capacity and release in reducing power by action on amylose was similar to that for commercial ?-amylase. Activities of ?-amylase up to 4.4 U ml?1 (1 U represents 1 ?mol of glucose equivalents released per min) were obtained for cultures grown on 2.5 mg of starch ml?1 in shaken cultures. No growth occurred in unshaken cultures. With elevated concentrations of starch (>2.5 mg ml?1), ?-amylase production declined and glucose accumulated in the cultures. Addition of glucose to cultures grown on low levels of starch, in which little glucose accumulated, suppressed ?-amylase production, and in bisubstrate growth studies, active production of the enzyme only occurred during growth on starch after glucose had been preferentially utilized. When cellulose, cellobiose, glucose, xylan, and xylose were tested as growth substrates for the production of ?-amylase (initial concentration, 2.5 mg ml?1), they were found to be less effective than starch, but maltose was almost as effective. The fungal ?-amylase was found to be stable at 60°C in the presence of low concentrations of starch (?5%), suggesting that it may be suitable for industrial application. PMID:16347742

Mountfort, Douglas O.; Asher, Rodney A.

1988-01-01

222

Potential role of lysozyme in bactericidal activity of in vitro-acquired salivary pellicle against Streptococcus faecium 9790.  

PubMed Central

The adherence of Streptococcus faecium 9790 to hydroxyapatite (HA) coated with whole saliva supernatant proteins (S-HA) or parotid fluid proteins was studied. The organism was labeled with [3H]thymidine, and adherence was estimated as the radioactivity remaining associated with the variously coated HA preparations after incubation and removal of unbound microbes by washing the adherence substratum. Adherence was time dependent and saturable, characteristics typical of oral streptococci in this in vitro adherence model system. However, adherence to S-HA, but not bare HA, was decreased 20-fold at 4 degrees C compared with room temperature. Furthermore, adherence at 4 degrees C to S-HA was decreased 20-fold relative to bare HA at 4 degrees C. Adherence to HA coated with parotid fluid proteins also was reduced at 4 degrees C. The magnitude of the temperature dependence and the inhibitory effect at 4 degrees C of whole saliva or parotid fluid pellicles on HA was unexpected. Of several sugars and amino sugars tested, the chitin saccharides, chitotriose, chitobiose, and N-acetylglucosamine caused greater than 90% inhibition of adherence to S-HA. These same saccharides were previously shown to inhibit lysozyme, polylysine, or autolytic lysis of the organism (N. J. Laible and G. R. Germaine, Infect. Immun. 48:720-728, 1985). Examination of unbound and adherent microbes revealed that lysis of the organism occurred during the adherence assays. A strong association (r = 0.83) between the extent of lysis and the extent of adherence was found under a variety of conditions. Depletion of lysozyme from saliva specimens used to coat HA resulted in a greater than 90% decrease in both cell lysis and adherence. Lysis of the microbe appeared dependent upon the presence of the saliva pellicle (coating) on HA, since solutions containing proteins desorbed from HA during mock-adherence incubations possessed lytic activity that was 2- to 10-fold too low to account for the extents of lysis observed with greater than or equal to 10(8) input cells. These results demonstrate the potential antibacterial activity of acquired salivary pellicle on enamel in vivo and the likely role of lysozyme in this activity. The data also serve to caution that this widely used in vitro adherence model will not distinguish whole-cell adherence from the adsorption of radiolabeled DNA released from lysing cells. Several additional controls are suggested that will indicate whether test microbes remain intact or lyse during adherence trials. PMID:3023239

Germaine, G R; Tellefson, L M

1986-01-01

223

Characterisation, immunolocalisation and antifungal activity of a lipid transfer protein from chili pepper (Capsicum annuum) seeds with novel ?-amylase inhibitory properties.  

PubMed

Lipid transfer proteins (LTPs) were thus named because they facilitate the transfer of lipids between membranes in vitro. This study was triggered by the characterization of a 9-kDa LTP from Capsicum annuum seeds that we call Ca-LTP(1) . Ca-LTP(1) was repurified, and in the last chromatographic purification step, propanol was used as the solvent in place of acetonitrile to maintain the protein's biological activity. Bidimensional electrophoresis of the 9-kDa band, which corresponds to the purified Ca-LTP(1) , showed the presence of three isoforms with isoelectric points (pIs) of 6.0, 8.5 and 9.5. Circular dichroism (CD) analysis suggested a predominance of ?-helices, as expected for the structure of an LTP family member. LTPs immunorelated to Ca-LTP(1) from C. annuum were also detected by western blotting in exudates released from C. annuum seeds and also in other Capsicum species. The tissue and subcellular localization of Ca-LTP(1) indicated that it was mainly localized within dense vesicles. In addition, isolated Ca-LTP(1) exhibited antifungal activity against Colletotrichum lindemunthianum, and especially against Candida tropicalis, causing several morphological changes to the cells including the formation of pseudohyphae. Ca-LTP(1) also caused the yeast plasma membrane to be permeable to the dye SYTOX green, as verified by fluorescence microscopy. We also found that Ca-LTP(1) is able to inhibit mammalian ?-amylase activity in vitro. PMID:21382036

Diz, Mariângela S; Carvalho, Andre O; Ribeiro, Suzanna F F; Da Cunha, Maura; Beltramini, Leila; Rodrigues, Rosana; Nascimento, Viviane V; Machado, Olga L T; Gomes, Valdirene M

2011-07-01

224

Effects of Physical Activity, Body Fat and Salivary Cortisol on Mucosal Immunity in Children  

Microsoft Academic Search

Abstract This study examined relationships among physical activity, body composition, and stress- and immunity-related variables in fifth grade children (10-11 yrs) in Southern Ontario. The 29 boys and 32 girls, who participated in the study, performed a 20m shuttle run for prediction of aerobic fitness. Bioelectrical impedance,was used to assess relative body fat (%BF). Standardized questionnaires were used to ,determine

Thomas J. Cieslak; Gail Frost; Panagiota Klentrou

2003-01-01

225

Thrombolytic Properties of Desmodus rotundus (vampire bat) Salivary Plasminogen Activator in Experimental Pulmonary Embolism in Rats  

Microsoft Academic Search

HE INITIAL anticipation that the safety of thrombo- T lytic therapy would improve with the introduction of human tissue-type plasminogen activator (t-PA) has not been fulfilled in clinical trials.' The expectation that bleed- ing episodes might be reduced with t-PA was based on early in vitro and experimental animal work which suggested that t-PA, unlike other clinically available thrombolytics, would

Werner Witt; Berthold Baldus; Peter Bringmann; Linda Cashion; Peter Donner; Wolf-Dieter Schleuning

1992-01-01

226

Salivary gland disorders.  

PubMed

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

Mandel, Louis

2014-11-01

227

Expression of ?-amylase in response to wounding in mung bean  

Microsoft Academic Search

The activity of a-amylase (EC 3.2.1.1) in mung bean (Vigna radiata (L.) Wilczek) cotyledons increased markedly in response to wounding. The changes in enzyme activity were in parallel with those in enzyme content. The level of a-amylase mRNA also notably increased in wounded cotyledons and attained its maximum level during the period between 1 and 2 d after wounding. The

Nobuya Koizuka; Yoshiyuki Tanaka; Yukio Morohashi

1995-01-01

228

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

PubMed Central

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

2014-01-01

229

A dual role of 20-hydroxyecdysone in the control of protein synthesis related to DNA puff activity in the anterior region of Bradysia hygida (Diptera, Sciaridae) salivary gland.  

PubMed

During the last 30 h of the larval stage, the salivary glands of Bradysia hygida show the amplification of some genes, resulting in the formation of two successive groups of DNA puffs, which direct the synthesis of two different sets of polypeptides. Incubation of anterior (S1) salivary gland regions, at age E7, beginning of first group of DNA puffs activity, in culture medium for 2 to 10 h results in a decrease in the synthesis of the polypeptides characteristic of this period. However, during subsequent incubation (from E7 to E7+12 h-24 h), when the second group of DNA puffs is active, S1 regions were able to synthesize some polypeptides characteristic of this period. The role of 20-OH ecdysone was studied, in vitro and in vivo, during these two periods of protein synthesis in S1 regions. The presence of the hormone was shown to be necessary to maintain, in vitro, the synthesis of the first set of polypeptides and was strongly inhibitory, in vitro and in vivo, to the synthesis of the second set of polypeptides. Thus, it is likely that the activity of the two distinct groups of DNA puffs is under opposite 20-OH-ecdysone control mechanisms. PMID:10844246

de Carvalho, D P; Coelho, P S; de Almeida, J C

2000-07-01

230

Modification of innervation pattern by fluoroquinolone treatment in the rat salivary glands.  

PubMed

Fluoroquinolone antibiotics (FQAs) are widely used in dental and medical therapy. Despite their known severe adverse actions on the central and peripheral nervous system, little attention has been directed toward the potential toxic side effects of these compounds on the oral tissues. As the saliva secretion is controlled by the nervous system and neuropeptides, the neurotoxic effect of pefloxacin (PEF), a representative member of FQAs, was studied in rats in the present work. Previously, we demonstrated a significant weight loss of parotid gland tissue, a marked decrease in 3H-thymidine incorporation, a decreased volume of saliva and amylase activity of the glandular tissue in response to PEF. Animals received intraperitoneal injection of PEF (20 mg/100 g body weight daily) for 3 and 7 days. Normal histology, and neurofilament 200, substance P (SP) and calcitonin gene-related polypeptide (CGRP) containing nerve fibers were detected with immunohistochemical methods. A marked decrease of the weights in salivary glands and the acinar diameters were measured. Similarly, a strong and significant decrease of the number of SP and CGRP containing nerve fibers were detected. These findings suggest that the impaired morphology and innervation pattern of salivary glands is related to the neurotoxic adverse effect of FQA treatment. PMID:19937634

Kelentey, Barna; Deak, Adam; Zelles, Tivadar; Matesz, Klara; Földes, Istvan; Veress, Gabor; Bacskai, Timea

2010-02-01

231

Transcriptome analysis of the salivary glands of potato leafhopper, Empoasca fabae.  

PubMed

The potato leafhopper, Empoasca fabae, is a pest of economic crops in the United States and Canada, where it causes damage known as hopperburn. Saliva, along with mechanical injury, leads to decreases in gas exchange rates, stunting and chlorosis. Although E. fabae saliva is known to induce plant responses, little knowledge exists of saliva composition at the molecular level. We subjected the salivary glands of E. fabae to Roche 454-pyrosequencing which resulted significant number (30,893) of expressed sequence tags including 2805 contigs and 28,088 singletons. A high number of sequences (78%) showed similarity to other insect species in GenBank, including Triboliumcastaneum, Drosophilamelanogaster and Acrythosiphonpisum. KEGG analysis predicted the presence of pathways for purine and thiamine metabolic, biosynthesis of secondary metabolites, drug metabolism, and lysine degradation. Pfam analysis showed a high number of cellulase and carboxylesterase protein domains. Expression analysis of candidate genes (alpha amylase, lipase, pectin lyase, etc.) among different tissues revealed tissue-specific expression of digestive enzymes in E. fabae. This is the first study to characterize the sialotranscriptome of E. fabae and the first for any species in the family of Cicadellidae. Due to the status of these insects as economic pests, knowledge of which genes are active in the salivary glands is important for understanding their impact on host plants. PMID:23063500

DeLay, Bridget; Mamidala, Praveen; Wijeratne, Asela; Wijeratne, Saranga; Mittapalli, Omprakash; Wang, Jian; Lamp, William

2012-12-01

232

Biochemical Characterization and Mass Spectrometric Disulfide Bond Mapping of Periplasmic -Amylase MalS of Escherichia coli*  

E-print Network

Biochemical Characterization and Mass Spectrometric Disulfide Bond Mapping of Periplasmic -Amylase Republic of Germany Periplasmic -amylase of Escherichia coli, the malS gene product, hydrolyzes linear-nitrophenylhexaoside hydrolyzed per min per mg of protein. Amylase activity was optimal at pH 8 and was dependent on divalent

Rippe, Karsten

233

Anopheles salivary gland proteomes from major malaria vectors  

PubMed Central

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

2012-01-01

234

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 ...

235

Stabilization of ?-amylase by using anionic surfactant during the immobilization process  

NASA Astrophysics Data System (ADS)

This work describes the entrapment of ?-amylase into butylacrylate-acrylic acid copolymer (BuA/AAc) using ? irradiation. The effect of an anionic surfactant (AOT), the reuse efficiency, and kinetic behavior of immobilized ?-amylase were studied. Covering of ?-amylase with bis-(2-ethylhexyl)sulfosuccinate sodium salt (AOT) made the enzyme more stable than the uncovered form. The hydrolytic activity of the pre-coated immobilized ?-amylase was increased below the critical micelle concentration (cmc) (10 mmol/L). The results showed an increase in the relative activity with increase in the degree of hydration. The pre-coated immobilized ?-amylase showed a higher k/K and lower activation energy compared to the free and uncoated-immobilized preparation, respectively. The results suggest that the immobilization of ?-amylase is a potentially useful approach for commercial starch hydrolysis in two-phase systems.

El-Batal, A. I.; Atia, K. S.; Eid, M.

2005-10-01

236

Evolutionary Studies on an ? -amylase Gene Segment in Bats and other Mammals  

Microsoft Academic Search

Comparative studies of salivary glands showed that they maybe related to the adaptive radiation of bats, especially in the\\u000a family Phylostomidae. In this study we have been searching for a likely relationship between different feeding habits found\\u000a in bats and possible adaptive changes in a coding segment of the ?-amylase enzyme. We have also tested some hypothesis about the phylogenetic

Rodrigo A. F. Redondo; Fabrício R. Santos

2006-01-01

237

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

PubMed

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

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

2001-04-10

238

An Endogenous ?-Amylase Inhibitor in Barley Kernels 1  

PubMed Central

Barley (Hordeum distichum cv Klages) kernels were shown to contain a factor that converted malted barley ?-amylase II to the ?-amylase III form. After purification by ammonium sulfate fractionation, ion exchange chromatography on DEAE-Sephacel, and gel-filtration on Bio Gel P60, the factor gave a single band of protein on isoelectric focusing. The purified factor inhibited hydrolysis of soluble starch by ?-amylase II from malted barley and germinated wheat (Triticum aestivum cv Neepawa). However, ?-amylase I from these cereals was not affected. The inhibitor was not dialyzable and was retained by a PM 10 ultrafiltration membrane suggesting a molecular weight greater than 10,000 daltons. Heat treatment of the inhibitor at 70°C for 15 minutes at pH 5.5 and 8.0 resulted in considerable loss of inhibitory activity. Images Fig. 1 Fig. 2 Fig. 3 PMID:16663089

Weselake, Randall J.; MacGregor, Alexander W.; Hill, Robert D.

1983-01-01

239

Original article Adaptative significance of amylase  

E-print Network

Original article Adaptative significance of amylase polymorphism in Drosophila. III. Geographic patterns in Drosophila subobscura tissue-specific expression of amylase in adult midgut M Andjelkovi controlled variation in tissue-specific expression of a-amylase enzyme. Polymorphism for amylase tissue

Paris-Sud XI, Université de

240

Immunohistochemical study of carbonic anhydrase in mixed tumours from major salivary glands and skin  

Microsoft Academic Search

Immunohistochemical distribution of carbonic anhydrase isoenzyme I and II was studied in mixed tumours of major salivary glands and skin. The normal salivary glands displayed strong carbonic anhydrase activity in both ductal epithelium and serous acinar cells and the serous demilune cells in the submandibular glands, including the eccrine ducts. Pleomorphic adenoma salivary gland origin exhibited positive staining in the

Yohko Noda; Yoshiaki Takai; Yoshimasa Iwai; Michael A. Meenaghan; Masahiko Mori

1986-01-01

241

Anti-HIV-1 activity of salivary MUC5B and MUC7 mucins from HIV patients with different CD4 counts  

PubMed Central

Background We have previously shown that MUC5B and MUC7 mucins from saliva of HIV negative individuals inhibit HIV-1 activity by 100% in an in vitro assay. The purpose of this subsequent study was to investigate whether MUC5B and MUC7 from saliva of HIV patients or with full blown AIDS had a similar inhibitory activity against the virus. Methods Salivary MUC5B and MUC7 from HIV patients with different CD4 counts (< 200, 200-400 and > 400) were incubated with HIV-1 prior to infection of the human T lymphoblastoid cell line (CEM SS cells). Cells were then cultured and viral replication was measured by a qualitative p24 antigen assay. The size, charge and immunoreactivity of mucins from HIV negative and positive individuals was also analysed by SDS-PAGE, Western blot and ELISA respectively. Results It was shown that irrespective of their CD4 counts both MUC5B and MUC7 from HIV patients, unlike the MUC5B and MUC7 from HIV negative individuals, did not inhibit HIV-1 activity. Size, charge and immunoreactivity differences between the mucins from HIV negative and positive individuals and among the mucins from HIV patients of different CD4 count was observed by SDS-PAGE, Western blot and ELISA. Conclusions Purified salivary mucins from HIV positive patients do not inhibit the AIDS virus in an in vitro assay. Although the reason for the inability of mucins from infected individuals to inhibit the virus is not known, it is likely that there is an alteration of the glycosylation pattern, and therefore of charge of mucin, in HIV positive patients. The ability to inhibit the virus by aggregation by sugar chains is thus diminished. PMID:20946627

2010-01-01

242

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

PubMed Central

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

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

2014-01-01

243

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

PubMed Central

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

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

2014-01-01

244

Purification and Characterization of Pea Epicotyl ?-Amylase 1  

PubMed Central

The most abundant ?-amylase (EC 3.2.1.2) in pea (Pisum sativum L.) was purified greater than 880-fold from epicotyls of etiolated germinating seedlings by anion exchange and gel filtration chromatography, glycogen precipitation, and preparative electrophoresis. The electrophoretic mobility and relative abundance of this ?-amylase are the same as that of an exoamylase previously reported to be primarily vacuolar. The enzyme was determined to be a ?-amylase by end product analysis and by its inability to hydrolyze ?-limit dextrin and to release dye from starch azure. Pea ?-amylase is an approximate 55 to 57 kilodalton monomer with a pl of 4.35, a pH optimum of 6.0 (soluble starch substrate), an Arrhenius energy of activation of 6.28 kilocalories per mole, and a Km of 1.67 milligrams per milliliter (soluble starch). The enzyme is strongly inhibited by heavy metals, p-chloromer-curiphenylsulfonic acid and N-ethylmaleimide, but much less strongly by iodoacetamide and iodoacetic acid, indicating cysteinyl sulfhydryls are not directly involved in catalysis. Pea ?-amylase is competitively inhibited by its end product, maltose, with a Ki of 11.5 millimolar. The enzyme is partially inhibited by Schardinger maltodextrins, with ?-cyclohexaamylose being a stronger inhibitor than ?-cycloheptaamylose. Moderately branched glucans (e.g. amylopectin) were better substrates for pea ?-amylase than less branched or non-branched (amyloses) or highly branched (glycogens) glucans. The enzyme failed to hydrolyze native starch grains from pea and glucans smaller than maltotetraose. The mechanism of pea ?-amylase is the multichain type. Possible roles of pea ?-amylase in cellular glucan metabolism are discussed. Images Figure 2 Figure 3 Figure 4 PMID:16667324

Lizotte, Pauline A.; Henson, Cynthia A.; Duke, Stanley H.

1990-01-01

245

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

Microsoft Academic Search

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

Yvonne R. Carter; Michael Ashburner

1972-01-01

246

Affinity column purification of amylases on protein inhibitors from wheat kernel.  

PubMed

An affinity column was devised for the purification of a large number of amylases inhibited by the albumin from wheat kernel. The procedure involved linking the protein inhibitors from wheat to Sepharose and then specifically eluting the amylase adsorbed to the gel with a high concentration of maltose. By this procedure, the amylases from Tenebrio molitor L. (yellow mealworm) larvae and chicken pancreas were purified to homogeneity with good yields for the first time, as shown by both alkaline and acidic electrophoresis. Human saliva alpha-amylase, purified by the same procedure, showed specific activity and electrophoretic patterns similar to those obtained by other workers with different techniques. PMID:241755

Buonocore, V; Poerio, E; Gramenzi, F; Silano, V

1975-11-12

247

Evaluation of Salivary Profile among Adult Type 2 Diabetes Mellitus Patients in South India  

PubMed Central

Background: A lack of consensus on the possible association between diabetes and salivary dysfunction motivated us to conduct this investigation on the salivary parameters in diabetic and non diabetic subjects. This could also make the use of saliva as an alternative to that of blood in the diagnosis/monitoring of diabetes mellitus. Objectives: To compare the salivary flow rates and the salivary physical and biochemical parameters of diabetic (D) and non diabetic (ND) subjects. Material and Methods: The participants in this study included 30 non diabetic subjects and 30 diabetic volunteers who had Type 2 Diabetes mellitus for a minimum of 2 years. Unstimulated whole saliva was collected in the fasting state. Salivary pH, flow rate and organic and inorganic constituents were evaluated. Data which was collected was statistically analysed and interpreted. Results: Salivary pH (ND=7.09±0.29, D=6.69±0.35), flow rate (ND=0.67±0.07, D=0.46±0.02) and salivary amylase (ND=92.51±13.74, D=19.20±1.8) were significantly lower in diabetics. They had significantly higher levels of salivary glucose (ND=4.33 ± 0.29, D=17.31±2.05), total proteins (ND=424.46±237.34, D=877.29±603.84), sodium (ND=4.31±0.65, D=14.42±1.83) and potassium (ND=20.84±0.71, D=25.95±1.56) and lower levels of calcium (ND=6.39±0.5, D=4.22±0.12) in comparison to those in the non-diabetic group. Conclusion: Significant variations were observed in salivary physical and biochemical parameters between diabetics and non diabetics. Evaluation of salivary parameters can be a cost effective and a non invasive alternative for screening, diagnosis and monitoring of diabetes, to blood. PMID:24086848

K.M, Prathibha; Johnson, Priscilla; Ganesh, Mathangi; Subhashini, Arcot S.

2013-01-01

248

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...

249

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

PubMed Central

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

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

250

Structural basis for the inhibition of mammalian and insect alpha-amylases by plant protein inhibitors.  

PubMed

Alpha-amylases are ubiquitous proteins which play an important role in the carbohydrate metabolism of microorganisms, animals and plants. Living organisms use protein inhibitors as a major tool to regulate the glycolytic activity of alpha-amylases. Most of the inhibitors for which three-dimensional (3-D) structures are available are directed against mammalian and insect alpha-amylases, interacting with the active sites in a substrate-like manner. In this review, we discuss the detailed inhibitory mechanism of these enzymes in light of the recent determination of the 3-D structures of pig pancreatic, human pancreatic, and yellow mealworm alpha-amylases in complex with plant protein inhibitors. In most cases, the mechanism of inhibition occurs through the direct blockage of the active center at several subsites of the enzyme. Inhibitors exhibiting "dual" activity against mammalian and insect alpha-amylases establish contacts of the same type in alternative ways. PMID:14871658

Payan, Françoise

2004-02-12

251

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

PubMed

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

Hyun, H H; Zeikus, J G

1985-12-01

252

Further Experiments on Gibberellin-Stimulated Amylase Production in Cereal Grains  

ERIC Educational Resources Information Center

Experiments conducted on wheat and barley grains to analyze activities of alpha- and beta-amylase enzymes. Gibberellins were used exogenously. Techniques are described in detail. Results on different cultivars revealed that beta-amylase was not an invariable result of imbibition. Techniques employed can be used by school students. (PS)

Coppage, Jo; Hill, T. A.

1973-01-01

253

Salivary Gland Changes in Disease  

Microsoft Academic Search

Ultrastructural alterations occurring in human salivary glands as a result of a variety of diseases are described. Major changes in these organs in cases of cystic fibrosis are probably the result of duct blockage, as indicated by study of chronically inflamed salivary glands. A new disease of salivary glands is reported in which parotid serous granules are distorted by bundles

B. Tandler

1987-01-01

254

The Heterochromatic Rolled Gene of Drosophila Melanogaster Is Extensively Polytenized and Transcriptionally Active in the Salivary Gland Chromocenter  

PubMed Central

This paper reports a cytogenetic and molecular study of the structural and functional organization of the Drosophila melanogaster chromocenter. The relations between mitotic (constitutive) heterochromatin and ?- and ?-heterochromatin are not fully understood. In the present work, we have studied the polytenization of the rolled (rl) locus, a 100-kb genomic region that maps to the proximal heterochromatin of chromosome 2 and has been previously thought to contribute to ?-heterochromatin. We show that rolled undergoes polytenization in salivary gland chromosomes to a degree comparable to that of euchromatic genes, despite its deep heterochromatic location. In contrast, both the Bari-1 sequences and the AAGAC satellite repeats, located respectively to the left and right of rl, are severely underrepresented and thus both appear to be ?-heterochromatic. In addition, we found that rl is transcribed in polytene tissues. Together, the results reported here indicate that functional sequences located within the proximal constitutive heterochromatin can undergo polytenization, contributing to the formation of ?-heterochromatin. The implications of this finding to chromocenter structure are discussed. PMID:8878678

Berghella, L.; Dimitri, P.

1996-01-01

255

Partial Purification and Characterization of a Thermostable Actinomycete ?-Amylase  

PubMed Central

A thermostable amylase, possibly a ?-amylase from Thermoactinomyces sp. no. 2 isolated from soil, is reported. The enzyme was purified 36-fold by acetone precipitation, ion-exchange chromatography, and Sephadex G-200 gel filtration, and the molecular weight was estimated at 31,600. The enzyme was characterized by demonstration of optimum activity at 60°C and pH 7 and by retention of 70% activity at 70°C (30 min). It was stimulated by Mn2+ and Fe2+ but strongly inhibited by Hg2+. Maltose was the only detectable product of hydrolysis of starches and was quantitatively highest in plantain starch hydrolysate. PMID:16346495

Obi, S. K. C.; Odibo, F. J. C.

1984-01-01

256

Amylase production by endophytic fungi Cylindrocephalumsp. isolated from medicinal plant Alpinia calcarata (Haw.) Roscoe  

PubMed Central

Amylases are among the most important enzymes used in modern biotechnology particularly in the process involving starch hydrolysis. Fungal amylase has large applications in food and pharmaceutical industries. Considering these facts, endophytic fungi isolated from the plant Alpinia calcarata (Haw.) Roscoe were screened for amylolytic activity on glucose yeast extract peptone agar (GYP) medium. Among thirty isolates of endophytic fungi, isolate number seven identified as Cylindrocephalum sp. (Ac-7) showed highest amylolytic activity and was taken for further study. Influence of various physical and chemical factors such as pH, temperature, carbon and nitrogen sources on amylase production in liquid media were studied. The maximal amylase production was found to be at 30ºC and at pH 7.0 of the growth medium. Among the various carbon and nitrogen sources tested, maltose at 1.5% and Sodium nitrate at 0.3% respectively gave optimum amylase production. PMID:24031946

Sunitha, V. H.; Ramesha, A.; Savitha, J.; Srinivas, C

2012-01-01

257

Full-fledged proteomic analysis of bioactive wheat amylase inhibitors by a 3-D analytical technique: Identification of new heterodimeric aggregation states.  

PubMed

Wheat proteinaceous alpha-amylase inhibitors (alpha-AIs) are increasingly investigated for their agronomical role as natural defence molecules of plants against the attack of insects and pests, but also for their effects on human health. The wheat genomes code for several bioactive alpha-AIs that share sequence homology, but differ in their specificity against alpha-amylases from different species and for their aggregation states. Wheat alpha-AIs are traditionally classified as belonging to the three classes of tetrameric, homodimeric and monomeric forms, each class being constituted by a number of polypeptides that display different electrophoretic mobilities. Here we describe a proteomic approach for the identification of bioactive alpha-AIs from wheat and, in particular, a 3-D technique that allows to best identify and characterize the dimeric fraction. The technique takes advantage of the thermal resistance of alpha-AIs (resistant to T > 70 degrees C) and consists in the separation of protein mixtures by 2-D polyacrylamide/starch electrophoresis under nondissociating PAGE (ND-PAGE, first dimension) and dissociating (urea-PAGE or U-PAGE second dimension) conditions, followed by in-gel spontaneous reaggregation of protein complexes and identification of the alpha-amylase inhibitory activity (antizymogram, third dimension) using enzymes from human salivary glands and from the larvae of Tenebrio molitor coleopter (yellow mealworm). Dimeric alpha-AIs from Triticum aestivum (bread wheat) were observed to exist as heterodimers. The formation of heterodimeric complexes was also confirmed by in vitro reaggregation assays carried out on RP-HPLC purified wheat dimeric alpha-AIs, and their bioactivity assayed by antizymogram analysis. The present 3-D analytical technique can be exploited for fast, full-fledged identification and characterization of wheat alpha-AIs. PMID:17203506

Zoccatelli, Gianni; Dalla Pellegrina, Chiara; Mosconi, Silvia; Consolini, Marica; Veneri, Gianluca; Chignola, Roberto; Peruffo, Angelo; Rizzi, Corrado

2007-02-01

258

Dose- and tissue-specific interaction of monoterpenes with the gibberellin-mediated release of potato tuber bud dormancy, sprout growth and induction of ?-amylases and ?-amylases.  

PubMed

Gibberellins (GA) are involved in bud dormancy release in several species. We show here that GA-treatment released bud dormancy, initiated bud sprouting and promoted sprout growth of excised potato tuber bud discs ('eyes'). Monoterpenes from peppermint oil (PMO) and S-(+)-carvone (CAR) interact with the GA-mediated bud dormancy release in a hormesis-type response: low monoterpene concentrations enhance dormancy release and the initiation of bud sprouting, whereas high concentrations inhibit it. PMO and CAR did, however, not affect sprout growth rate after its onset. We further show that GA-induced dormancy release is associated with tissue-specific regulation of ?- and ?-amylases. Molecular phylogenetic analysis shows that potato ?-amylases cluster into two distinct groups: ?-AMY1 and ?-AMY2. GA-treatment induced transcript accumulation of members of both ?-amylase groups, as well as ?- and ?-amylase enzyme activity in sprout and 'sub-eye' tissues. In sprouts, CAR interacts with the GA-mediated accumulation of ?-amylase transcripts in an ?-AMY2-specific and dose-dependent manner. Low CAR concentrations enhance the accumulation of ?-AMY2-type ?-amylase transcripts, but do not affect the ?-AMY1-type transcripts. Low CAR concentrations also enhance the accumulation of ?- and ?-amylase enzyme activity in sprouts, but not in 'sub-eye' tissues. In contrast, high CAR concentrations have no appreciable effect in sprouts on the enzyme activities and the ?-amylase transcript abundances of either group. The dose-dependent effects on the enzyme activities and the ?-AMY2-type ?-amylase transcripts in sprouts are specific for CAR but not for PMO. Different monoterpenes therefore may have specific targets for their interaction with hormone signalling pathways. PMID:21858448

Rentzsch, Sonja; Podzimska, Dagmara; Voegele, Antje; Imbeck, Madeleine; Müller, Kerstin; Linkies, Ada; Leubner-Metzger, Gerhard

2012-01-01

259

Purification and characterization of thermostable ?-amylase from thermophilic Anoxybacillus flavithermus.  

PubMed

This study reports on the purification and characterization of thermostable ?-amylase (?-1-4 D-glucan glucanohydrolase EC 3.2.1.1) from a newly isolated Anoxybacillus flavithermus. A. flavithermus was used, which was isolated from hot water springs of Ömer, Afyonkarahisar, Turkey. The gram-positive, spore-forming, motile, moderately thermophilic bacteria were found to be a strain of A. flavithermus analysed by 16S rRNA comparison. The optimal conditions for bacterial growth were determined to be at 20 thh, 55 °C and pH 6.0. Maximum ?-amylase activity was obtained at 55 °C at pH 7.0 after 24h of incubation. Thermostable ?-amylase from A. flavithermus was purified by 70% (NH4)2SO4 and ion-exchange chromatography (5.2-fold; 65.8% yield). The molecular weight of ?-amylase was 60 kDa, as estimated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The ?-amylase hydrolyzed soluble starch at 55 °C with Km: 0.005 mM and Vmax: 3.5 ?mol min(-1). PMID:24507266

Agülo?lu Fincan, S; Enez, B; Özdemir, S; Matpan Bekler, F

2014-02-15

260

Alpha-amylase from the Hyperthermophilic Archaeon Thermococcus thioreducens  

NASA Technical Reports Server (NTRS)

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.

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

2003-01-01

261

High-efficiency, one-step starch utilization by transformed Saccharomyces cells which secrete both yeast glucoamylase and mouse alpha-amylase.  

PubMed Central

Transformed, hybrid Saccharomyces strains capable of simultaneous secretion of glucoamylase and alpha-amylase have been produced. These strains could carry out direct, one-step assimilation of starch, with conversion efficiency greater than 93% during a 5-day growth period. One of the transformants converted 92.8% of available starch into reducing sugars in only 2 days. Glucoamylase secretion by these strains resulted from expression of one or more chromosomal STA genes derived from Saccharomyces diastaticus. The strains were transformed by a plasmid (pMS12) containing mouse salivary alpha-amylase cDNA in an expression vector containing yeast alcohol dehydrogenase promoter and a segment of yeast 2 micron plasmid. The major starch hydrolysis product produced by crude amylases found in culture broths was glucose, indicating that alpha-amylase and glucoamylase acted cooperatively. PMID:3132104

Kim, K; Park, C S; Mattoon, J R

1988-01-01

262

Properties of an amylase from thermophilic Bacillus SP  

PubMed Central

?-Amylase production by thermophilic Bacillus sp strain SMIA-2 cultivated in liquid cultures containing soluble starch as a carbon source and supplemented with 0.05% whey protein and 0.2% peptone reached a maximum activity at 32 h, with levels of 37 U/mL. Studies on the amylase characterization revealed that the optimum temperature of this enzyme was 90°C. The enzyme was stable for 1 h at temperatures ranging from 40-50°C while at 90°C, 66% of its maximum activity was lost. However, in the presence of 5 mM CaCl2, the enzyme was stable at 90°C for 30 min and retained about 58% residual activity after 1 h. The optimum pH of the enzyme was found to be 8.5. After incubation of enzyme for 2 h at pH 9.5 and 11.0 was observed a decrease of about 6.3% and 16.5% of its original activity. At pH 6.0 the enzyme lost about 36% of its original activity. The enzyme was strongly inhibited by Co2+, Cu2+ and Ba2+, but less affected by Mg2+, Na+ and K+. In the presence of 2.0 M NaCl, 63% of amylase activity was retained after 2 h incubation at 45°C. The amylase exhibited more than 70% activity when incubated for 1 h at 50°C with sodium dodecyl sulphate. However, very little residual activity was obtained with sodium hypochlorite and with hydrogen peroxide the enzyme was completely inhibited. The compatibility of Bacillus sp SMIA-2 amylase with certain commercial detergents was shown to be good as the enzyme retained 86%, 85% and 75% of its activity after 20 min incubation at 50°C in the presence of the detergent brands Omo®, Campeiro® and Tide®, respectively. PMID:24031188

de Carvalho, Raquel Vieira; Côrrea, Thamy Lívia Ribeiro; da Silva, Júlia Caroline Matos; de Oliveira Mansur, Luciana Ribeiro Coutinho; Martins, Meire Lelis Leal

2008-01-01

263

Establishment of Functional Acinar-like Cultures from Human Salivary Glands.  

PubMed

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

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

2015-02-01

264

Carbon nanotube-based electrochemical sensor for assay of salivary cholinesterase enzyme activity: an exposure biomarker of organophosphate pesticides and nerve agents.  

PubMed

Certain saliva enzymes may be useful biomarkers for detecting exposures to organophosphate pesticides and chemical nerve agents. In this regard, saliva biomonitoring offers a simple and noninvasive approach for rapidly evaluating those exposures in real time. An electrochemical sensor coupled with a microflow injection system was developed for a simple, rapid, and sensitive characterization of cholinesterase (ChE) enzyme activities in rat saliva. The electrochemical sensor is based on a carbon nanotube (CNT)-modified screen-printed carbon electrode (SPE), which is integrated into a flow cell. Because of the excellent electrocatalytic activity of the CNTs, the sensor can detect electroactive species that are produced from enzymatic reactions with extremely high sensitivity and at low potentials. The electrochemical properties of acetylcholinesterase (AChE) enzymatic products were studied using a CNT-modified SPE, and the operation parameters such as the applied potential and substrate concentration were optimized to achieve the best performance. The AChE enzyme activity was further investigated using the CNT-based electrochemical sensor with commercially available purified AChE and ChE in saliva obtained from nave rats. It is found that the calibration curve is linear over a wide range of AChE concentrations from 5 pM to 0.5 nM, and the sensor is very sensitive with the detection limit down to 2 pM. The dynamics of the ChE enzyme activity in saliva with organophosphate pesticides was further studied using this sensor. The results showthatthe senor can be used to characterize salivary enzyme activity and to detect the exposure to organophosphate compounds. This new CNT-based electrochemical sensor thus provides a sensitive and quantitative tool for noninvasive biomonitoring of the exposure to organophosphate pesticides and nerve agents. PMID:18505017

Wang, Jun; Timchalk, Charles; Lin, Yuehe

2008-04-01

265

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

PubMed

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

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

2014-01-01

266

Conformational stability and integrity of ?-amylase from mung beans: Evidence of kinetic intermediate in GdmCl-induced unfolding  

Microsoft Academic Search

?-Amylase from mung beans (Vigna radiata) being one of the few plant ?-amylases purified so far was studied with respect to its conformational stability by CD and fluorescence spectroscopy. The enzyme was shown to bind 3–4 Ca2+ ions, which all are important for its activity. In contrast to other ?-amylases no inhibition was observed at high Ca2+ concentrations (100 mM). Depletion

Pallavi Tripathi; Hagen Hofmann; Arvind M. Kayastha; Renate Ulbrich-Hofmann

2008-01-01

267

Rle de la muqueuse duodnale dans l'adaptation de l'?-amylase pancratique au rgime alimentaire chez le Porc  

E-print Network

Rôle de la muqueuse duodénale dans l'adaptation de l'?-amylase pancréatique au régime. The role of the duodenal mucosa in the adaptation of pancreatic a-amylase to the diet in the pig.001), total protein output (― 54.2 p. 100 ; P amylase activity (―15.8 p

Paris-Sud XI, Université de

268

A wireless magnetoelastic ?-amylase sensor  

Microsoft Academic Search

A remote query, inexpensive and hence readily disposable ?-amylase biosensor is described. The sensor is fabricated by immobilizing a layer of starch gel on a thick-film ribbon-like magnetoelastic sensor. In response to an ac magnetic field the magnetoelastic sensor replies with a return magnetic field, the characteristic resonance frequency of which is inversely dependent upon the attached mass load of

Shihui Wu; Yi Zhu; Qingyun Cai; Kefeng Zeng; Craig A. Grimes

2007-01-01

269

Management of Salivary Gland Cancer  

Microsoft Academic Search

\\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

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

270

Basic fibroblast growth factor in rat salivary glands  

Microsoft Academic Search

We studied the occurrence and localization of basic fibroblast growth factor (bFGF) in rat salivary glands using a specific monoclonal antibody. It was shown that the extract of rat salivary glands has a pronounced stimulatory activity on the growth of bovine capillary endothelial cells, which is blocked by the addition of an antibody against bFGF. The concentration of bFGF in

Osamu Amano; Yoshino Yoshitake; Katsuzo Nishikawa; Shoichi Iseki

1993-01-01

271

Taxonomy of Salivary Gland Neoplasm  

PubMed Central

Classification of neoplasms of any organ should be predicted on the patterns of differentiation that reflect the organization and cell types of the parental tissue. The ability to classify a neoplasm instills confidence in its predicted biologic behavior and the selection of treatment. There has not been a single universally used classification system for salivary gland tumor. Histogenetic and morphogenetic concepts and the developing information on various molecular parameters will have significant influence on the classification of salivary glands tumors. In this article we would highlight the histogenetic and morphogenetic concepts in salivary gland neoplasms and elaborate on the taxonomic system of classification of salivary gland neoplasms. PMID:24783163

Sreeja, C.; Shahela, Tanveer; Aesha, Syeda; Satish, Muthu Kumar

2014-01-01

272

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

PubMed

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

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

2014-10-01

273

59 FR- ]-Amylase Enzyme Preparation; Affirmation of GRAS Status as Direct Human Food Ingredient  

Federal Register 2010, 2011, 2012, 2013

...final reports and published articles of safety studies with -amylase...International Union of Biochemistry Enzyme Commission (E.C...CRC Critical Reviews in Biochemistry and Molecular Biology, 24...Active Sites,'' Journal of Biochemistry, 98:95-103,...

1994-12-05

274

Inhibition of alpha-glucosidase and amylase by bartogenic acid isolated from Barringtonia racemosa Roxb. seeds.  

PubMed

Barringtonia racemosa presents a wide range of therapeutic applications. In the course of identifying bioactives from Indian medicinal plants it was observed that the hexane, ethanol and methanol extracts of B. racemosa seeds displayed potent yeast and intestinal alpha-glucosidase inhibitory activities. The methanol extract was found to be superior among them. However, none of the extracts exhibited pancreatic alpha-amylase inhibitory activity, rather the ethanol and methanol extracts accelerated the alpha-amylase enzyme activity. Interestingly, however, bartogenic acid isolated from the methanol extract inhibited alpha-amylase also. This is the first report identifying alpha-glucosidase inhibitory activity in B. racemosa seed extracts and assigning to bartogenic acid an alpha-glucosidase and amylase inhibitory property. The presence of bartogenic acid in B. racemosa seeds as a major compound is also reported for the first time in this communication. PMID:17533638

Gowri, P Mangala; Tiwari, Ashok K; Ali, A Zehra; Rao, J Madhusudana

2007-08-01

275

Role of the non-catalytic triad in alpha-amylases.  

E-print Network

??La triade non-catalytique est un motif strictement conservé des alpha-amylases chlorure-dépendentes qui est parfaitement superposable avec la triade catalytique des protéases à sérine active. Le… (more)

Marx, Jean-Claude

2007-01-01

276

Purification of extrachloroplastic. beta. -amylase from leaves of starchless and wild type Arabidopsis  

SciTech Connect

Amylase activity in crude leaf extracts from starchless mutants of Arabidopsis thaliana is 5 to 10 fold higher than in the wild type (WT) when plants are grown under a 12 h photoperiod. Visualized on native PAGE, the increased activity is attributed primarily to a previously characterized extrachloroplastic {beta}-(exo)amylase. The {beta}-amylases from phosoglucomutase deficient (starchless) and WT leaves were purified to homogeneity in two steps utilizing polyethylene glycol fractionation, and cyclohexaamylose affinity chromatography. The enzyme from both mutant and WT leaves had negligible activity toward either {beta}-limit dextrin or pullulan. The specific activities of both purified enzymes were similar indicating that the protein is over-expressed in the mutant. Preliminary antibody neutralization experiments suggest that the two {beta}-amylases are not different.

Somerville, C.; Monroe, J.; Preiss, J. (Michigan Sate Univ., East Lansing (USA))

1989-04-01

277

Suppression of ?-amylase genes improves quality of rice grain ripened under high temperature.  

PubMed

High temperature impairs rice (Oryza sativa) grain filling by inhibiting the deposition of storage materials such as starch, resulting in mature grains with a chalky appearance, currently a major problem for rice farming in Asian countries. Such deterioration of grain quality is accompanied by the altered expression of starch metabolism-related genes. Here we report the involvement of a starch-hydrolyzing enzyme, ?-amylase, in high temperature-triggered grain chalkiness. In developing seeds, high temperature induced the expression of ?-amylase genes, namely Amy1A, Amy1C, Amy3A, Amy3D and Amy3E, as well as ?-amylase activity, while it decreased an ?-amylase-repressing plant hormone, ABA, suggesting starch to be degraded by ?-amylase in developing grains under elevated temperature. Furthermore, RNAi-mediated suppression of ?-amylase genes in ripening seeds resulted in fewer chalky grains under high-temperature conditions. As the extent of the decrease in chalky grains was highly correlated to decreases in the expression of Amy1A, Amy1C, Amy3A and Amy3B, these genes would be involved in the chalkiness through degradation of starch accumulating in the developing grains. The results show that activation of ?-amylase by high temperature is a crucial trigger for grain chalkiness and that its suppression is a potential strategy for ameliorating grain damage from global warming. PMID:22967050

Hakata, Makoto; Kuroda, Masaharu; Miyashita, Tomomi; Yamaguchi, Takeshi; Kojima, Mikiko; Sakakibara, Hitoshi; Mitsui, Toshiaki; Yamakawa, Hiromoto

2012-12-01

278

Characteristics of Two Forms of ?-Amylases and Structural Implication  

PubMed Central

Complete (Ba-L) and truncated (Ba-S) forms of ?-amylases from Bacillus subtilis X-23 were purified, and the amino- and carboxyl-terminal amino acid sequences of Ba-L and Ba-S were determined. The amino acid sequence deduced from the nucleotide sequence of the ?-amylase gene indicated that Ba-S was produced from Ba-L by truncation of the 186 amino acid residues at the carboxyl-terminal region. The results of genomic Southern analysis and Western analysis suggested that the two enzymes originated from the same ?-amylase gene and that truncation of Ba-L to Ba-S occurred during the cultivation of B. subtilis X-23 cells. Although the primary structure of Ba-S was approximately 28% shorter than that of Ba-L, the two enzyme forms had the same enzymatic characteristics (molar catalytic activity, amylolytic pattern, transglycosylation ability, effect of pH on stability and activity, optimum temperature, and raw starch-binding ability), except that the thermal stability of Ba-S was higher than that of Ba-L. An analysis of the secondary structure as well as the predicted three-dimensional structure of Ba-S showed that Ba-S retained all of the necessary domains (domains A, B, and C) which were most likely to be required for functionality as ?-amylase. PMID:10508102

Ohdan, Kohji; Kuriki, Takashi; Kaneko, Hiroki; Shimada, Jiro; Takada, Toshikazu; Fujimoto, Zui; Mizuno, Hiroshi; Okada, Shigetaka

1999-01-01

279

Beta-amylase in germinating millet seeds.  

PubMed

Beta-amylase (EC 3.2.1.2) was isolated from germinating millet (Panicum miliaceum L.) seeds by a procedure that included ammonium sulfate fractionation, chromatography on DEAE-cellulofine and CM-cellulofine, and preparative isoelectric focusing. The enzyme was homogeneous by SDS-PAGE. The M(r) of the enzyme was estimated to be 58,000 based on its mobility on SDS-PAGE and gel filtration with TSKgel G4000SW(XL), which showed that it is composed of a single unit. The isoelectric point of the enzyme was 4.62. The enzyme hydrolyzed malto-oligosaccharides more readily as their degree of polymerization increased, this being strongest for malto-oligosaccharides larger than 13 glucose residues and very weakly for maltotriose. Amylose, amylopectin and soluble starch were the most suitable substrates for the enzyme. While the enzyme showed some activity against native starch by itself, starch digestion was accelerated 2.5-fold using alpha-amylase, pullulanase and alpha-glucosidase. This enzyme appears to be very important for the germination of millet seeds. PMID:14561508

Yamasaki, Yoshiki

2003-11-01

280

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

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

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

2010-01-01

281

Salivary gland diseases in children  

PubMed Central

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

Iro, Heinrich; Zenk, Johannes

2014-01-01

282

What Is Salivary Gland Cancer?  

MedlinePLUS

... grow into or spread to the salivary glands. Non-Hodgkin lymphoma: Most non-Hodgkin lymphomas start in lymph nodes. Rarely, these cancers start ... For more information on lymphomas, see our document Non-Hodgkin Lymphoma . Sarcomas: The salivary glands contain blood vessels, muscle ...

283

Induction of a Peptide with Activity against a Broad Spectrum of Pathogens in the Aedes aegypti Salivary Gland, following Infection with Dengue Virus  

Microsoft Academic Search

The ultimate stage of the transmission of Dengue Virus (DENV) to man is strongly dependent on crosstalk between the virus and the immune system of its vector Aedes aegypti (Ae. aegypti). Infection of the mosquito's salivary glands by DENV is the final step prior to viral transmission. Therefore, in the present study, we have determined the modulatory effects of DENV

Natthanej Luplertlop; Pornapat Surasombatpattana; Sirilaksana Patramool; Emilie Dumas; Ladawan Wasinpiyamongkol; Laure Saune; Rodolphe Hamel; Eric Bernard; Denis Sereno; Frédéric Thomas; David Piquemal; Hans Yssel; Laurence Briant; Dorothée Missé

2011-01-01

284

21 CFR 862.1070 - Amylase test system.  

...2014-04-01 2014-04-01 false Amylase test system. 862.1070 Section 862...Clinical Chemistry Test Systems § 862.1070 Amylase test system. (a) Identification. An amylase test system is a device intended to...

2014-04-01

285

21 CFR 862.1070 - Amylase test system.  

Code of Federal Regulations, 2011 CFR

...2011-04-01 2011-04-01 false Amylase test system. 862.1070 Section 862...Clinical Chemistry Test Systems § 862.1070 Amylase test system. (a) Identification. An amylase test system is a device intended to...

2011-04-01

286

21 CFR 862.1070 - Amylase test system.  

Code of Federal Regulations, 2010 CFR

...2010-04-01 2010-04-01 false Amylase test system. 862.1070 Section 862...Clinical Chemistry Test Systems § 862.1070 Amylase test system. (a) Identification. An amylase test system is a device intended to...

2010-04-01

287

21 CFR 862.1070 - Amylase test system.  

Code of Federal Regulations, 2012 CFR

...2012-04-01 2012-04-01 false Amylase test system. 862.1070 Section 862...Clinical Chemistry Test Systems § 862.1070 Amylase test system. (a) Identification. An amylase test system is a device intended to...

2012-04-01

288

21 CFR 862.1070 - Amylase test system.  

Code of Federal Regulations, 2013 CFR

...2013-04-01 2013-04-01 false Amylase test system. 862.1070 Section 862...Clinical Chemistry Test Systems § 862.1070 Amylase test system. (a) Identification. An amylase test system is a device intended to...

2013-04-01

289

An exceptionally cold-adapted alpha-amylase from a metagenomic library of a cold and alkaline environment.  

PubMed

A cold-active ?-amylase, AmyI3C6, identified by a functional metagenomics approach was expressed in Escherichia coli and purified to homogeneity. Sequence analysis showed that the AmyI3C6 amylase was similar to ?-amylases from the class Clostridia and revealed classical characteristics of cold-adapted enzymes, as did comparison of the kinetic parameters K m and k cat to a mesophilic ?-amylase. AmyI3C6 was shown to be heat-labile. Temperature optimum was at 10-15 °C, and more than 70 % of the relative activity was retained at 1 °C. The pH optimum of AmyI3C6 was at pH 8-9, and the enzyme displayed activity in two commercial detergents tested, suggesting that the AmyI3C6 ?-amylase may be useful as a detergent enzyme in environmentally friendly, low-temperature laundry processes. PMID:25038927

Vester, Jan Kjølhede; Glaring, Mikkel Andreas; Stougaard, Peter

2014-07-20

290

Wheat grain preharvest sprouting and late maturity alpha-amylase.  

PubMed

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

Mares, Daryl J; Mrva, Kolumbina

2014-12-01

291

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

PubMed

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

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

2015-02-01

292

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

PubMed

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

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

293

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

PubMed Central

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

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

294

Molecular cloning and characterization of amylase from soil metagenomic library derived from Northwestern Himalayas  

Microsoft Academic Search

The increasing demand for novel biocatalysts stimulates exploration of resources from soil. Metagenomics, a culture independent\\u000a approach, represent a sheer unlimited resource for discovery of novel biocatalysts from uncultured microorganisms. In this\\u000a study, a soil-derived metagenomic library containing 90,700 recombinants was constructed and screened for lipase, cellulase,\\u000a protease and amylase activity. A gene (pAMY) of 909 bp encoding for amylase was

Sarika Sharma; Farrah Gul Khan; Ghulam Nabi Qazi

2010-01-01

295

Control of ?-Amylase Development in Cotyledons during and following Germination of Mung Bean Seeds 1  

PubMed Central

Developmental patterns of ?-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, ?-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, ?-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 ?-amylase. We propose that the regulation of the development of ?-amylase in cotyledons may involve some factor(s) inhibitory to accumulation of ?-amylase mRNA, which is present in dry cotyledons and can be removed from cotyledons by leakage or by the presence of the axis. Images Figure 3 Figure 5 Figure 6 PMID:16667006

Morohashi, Yukio; Katoh, Hisashi; Kaneko, Yasuko; Matsushima, Hisashi

1989-01-01

296

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

PubMed

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

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

2014-11-20

297

Salivary alterations in diabetes mellitus.  

PubMed

Separately collected parotid and submaxillary salivary samples from 20 diabetic and 20 matched control subjects were analyzed for flow rate, electrolyte content and immunoglobulin (IgA, IgG and IgM) levels. Flow rates did not vary significantly between the two groups of subjects; calcium ion content, however, was higher in the diabetic subjects for both salivary glands. The presence of salivary IgI in 6 of 10 patients was also a significant finding. Any attempt to draw a conclusion between the severity of periodontal disease and Diabetes Mellitus from the above findings is still speculative but does indicate further areas of research. PMID:1057651

Marder, M Z; Abelson, D C; Mandel, I D

1975-09-01

298

In vitro ? -amylase and ?-glucosidase inhibitory potential of Trigonella foenum-graecum leaves extract.  

PubMed

Trigonella foenum-graecum is one of the widely used herbs in food and medicine. The seeds of the plants are investigated for antidiabetic potential; however, no efforts have been done to explore the potential of leaves to modify carbohydrate metabolizing enzymes viz. ?-amylase and ?-glucosidase. The present work was designed to investigate the inhibitory potential of ethyl acetate and water extract of T. foenum-graecum on enzymes ?-amylase and ?-glucosidase. Different concentrations of extracts were used to study inhibition of enzymatic activity of ?-amylase and ?-glucosidase. A dose dependent inhibitory effect on enzymes was observed. The current study, for the first time, revealed ?-amylase and ?-glucosidase inhibitory potential of T. foenum-graecum and the study could be helpful to isolate and characterize compounds responsible for it. PMID:24049415

Ganeshpurkar, Aditya; Diwedi, Varsha; Bhardwaj, Yash

2013-01-01

299

Lysosomal and Free Acid Phosphatase in Salivary Glands of Chironomus tentans  

Microsoft Academic Search

In cells of salivary glands of last-instar larvae of Chironomus tentans, acid phosphatase activity is bound to (probable) lysosomes and a few other cell organelles. At the end of the pupal molt the salivary gland breaks down. While acid phosphatase in areas of nondegenerated cells is still restricted to the structures mentioned, in degenerated areas the enzyme is freely distributed

Ki Ssu Schin; Ulrich Clever

1965-01-01

300

Physical and catalytic properties of alpha-amylase from Tenebrio molitor L. larvae.  

PubMed

The amylase from Tenebrio molitor L. larvae (yellow mealworm) was characterized according to a number of its molecular and catalytic properties. The insect amylase is a single polypeptide chain with mol.wt. 68000, an isoelectric point of 4.0 and a very low content of sulphur-containing amino acids. The enzyme is a Ca2+-protein and behaves as an alpha-amylase. Removal of Ca2+ by exhaustive dialysis against water causes the irreversible inactivation of the enzyme. Moreover, the enzyme is activated by the presence in the assay mixture of Cl-, or some other inorganic anions that are less effective than Cl-, and is inhibited by F-. Optimal conditions of pH and temperature for the enzymic activity are 5.8 and 37 degrees C. The insect amylase exhibits an identical kinetic behaviour toward starch, amylose and amylopectin; the enzyme hydrolyses glycogen with a higher affinity constant. Compared with the non-insect alpha-amylases described in the literature, Tenebrio molitor amylase has a lower affinity for starch. PMID:942374

Buonocore, V; Poerio, E; Silano, V; Tomasi, M

1976-03-01

301

Optimization of alpha-amylase production for the green synthesis of gold nanoparticles.  

PubMed

Biocompatible gold nanoparticles have received considerable attention in recent years because of their promising applications in bioimaging, biosensors, biolabels, and biomedicine. The generation of gold nanoparticles using extra-cellular alpha-amylase for the reduction of AuCl(4) with the retention of enzymatic activity in the complex is being reported. The enhanced synthesis of particles has been brought about by optimizing the medium components for alpha-amylase. Response surface methodology and central composite rotary design (CCRD) were employed to optimize a fermentation medium for the production of alpha-amylase by Bacillus licheniformis at pH 8. The three variables involved in the study of alpha-amylase were fructose, peptone and soya meal. Only fructose had a significant effect on alpha-amylase production. The most optimum medium (medB) containing (%) fructose: 3, peptone: 1, soya meal: 2, resulted in a amylase activity of 201.381 U/ml which is same as that of the central level. The least optimum (medA) and most optimum (medB) media were compared for the synthesis of particles indicated by difference in color formation. Spectrophotometric analysis revealed that the particles exhibited a peak at 582 nm and the A(582) for the Med B was 8-fold greater than that of the Med A. The TEM analysis revealed that the particle size ranged from 10 to 50 nm. PMID:20189782

Kalishwaralal, Kalimuthu; Gopalram, Shubaash; Vaidyanathan, Ramanathan; Deepak, Venkatraman; Pandian, Sureshbabu Ram Kumar; Gurunathan, Sangiliyandi

2010-06-01

302

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

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

Lisi, Sabrina; Sisto, Margherita; D'Amore, Massimo; Lofrumento, Dario Domenico

2014-11-01

303

Production of raw-starch-digesting ?-amylase isoform from Bacillus sp. under solid-state fermentation and biochemical characterization.  

PubMed

?-Amylase production by solid-state fermentation of different Bacillus sp. was studied previously on different fermentation media. However, no study has been reported on the influence of selected media on expression of desired amylase isoforms such as raw-starch-digesting amylase (RSDA). In this paper, the influence of different inexpensive and available agro-resources as solid media (corn, wheat and triticale) on ?-amylase isoform induction from three wild-type Bacillus sp., selected among one hundred strains tested, namely 9B, 12B and 24A was investigated. For all three strains, tested amylases were detected in the multiple forms; however, number and intensity of each form differed depending on the solid media used for growth. To determine which isoform from Bacillus sp. 12B was RSDA, the suspected isoform was purified. The optimum pH for the purified ?-amylase isoform was 6.0-8.0, while the optimum temperature was 60-90 °C. Isoform was considerably thermostable and Ca(2+)-independent, and actually the only ?-amylase active towards raw starch. Purification and characterization of RSDA showed that not all of the solid media tested induced RSDA. From an economic point of view, it might be significant to obtain pure isoenzyme for potential use in the raw-starch hydrolysis, since it was 5 times more efficient in raw corn starch hydrolysis than the crude amylase preparation. PMID:24385152

Boži?, Nataša; Slavi?, Marinela Šokarda; Gavrilovi?, Anja; Vuj?i?, Zoran

2014-07-01

304

Purification of a. beta. -amylase that accumulates in Arabidopsis thaliana mutants defective in starch metabolism. [Arabidopsis thaliana  

SciTech Connect

Amylase activity is elevated 5- to 10-fold in leaves of several different Arabidopsis thaliana mutants defective in starch metabolism when they are grown under a 12-hour photoperiod. Activity is also increased when plants are grown under higher light intensity. It was previously determined that the elevated activity was an extrachloroplastic {beta}-(exo)amylase. Due to the location of this enzyme outside the chloroplast, its function is not known. The enzyme was purified to homogeneity from leaves of both a starchless mutant deficient in plastid phosphoglucomutase and from the wild type using polyethylene glycol fractionation and cyclohexaamylose affinity chromatography. The molecular mass of the {beta}-amylase from both sources was 55,000 daltons as determined by denaturing gel electrophoresis. Gel filtration studies indicated that the enzyme was a monomer. The specific activities of the purified protein from mutant and wild-type sources, their substrate specificities, and K{sub m} for amylopectin were identical. Based on these results it was concluded that the mutant contained an increased level of {beta}-amylase protein. Enzyme neutralization studies using a polyclonal antiserum raised to purified {beta}-amylase showed that in each of two starchless mutants, one starch deficient mutant and one starch overproducing mutant, the elevated amylase activity was due to elevated {beta}-amylase protein.

Monroe, J.D.; Preiss, J. (Michigan State Univ., East Lansing (USA))

1990-11-01

305

Cloning Sequencing and Expression of the Gene Encoding Extracellular a-Amylase from Pyrococcus furiosus and Biochemical Characterization of the Recombinant Enzyme  

Microsoft Academic Search

The gene encoding the hyperthermophilic extracellular a-amylase from Pyrococcus furiosus was cloned by activity screening in Escherichia coli. The gene encoded a single 460-residue polypeptide chain. The polypeptide contained a 26-residue signal peptide, indicating that this Pyrococcus a-amylase was an extracellular enzyme. Unlike the P. furiosus intracellular a-amylase, this extracellular enzyme showed 45 to 56% similarity and 20 to 35%

Guoqiang Dong; Claire Vieille; Alexei Savchenko; J. Gregory Zeikus

1997-01-01

306

The pathobiology of salivary gland  

Microsoft Academic Search

Summary In the transgenic TG.SH (mouse mammary tumour virus\\/v-Ha-ras) mouse, designed to develop mammary tumours, occasional spontaneous salivary gland tumours have been reported, predominantly in males. The incidence and histomorphology of salivary gland tumours in 73 TG.SH mice were surveyed and in total, 21.9% developed both overt and microscopic parotid tumours. The majority developed between 73 and 150 days of

Irving Dardick; Aileen P. Burford-Mason; David S. Garlick; Walter P. Carney

1992-01-01

307

Salivary Gland Cancer Stem Cells  

PubMed Central

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

Adams, April; Warner, Kristy; Nör, Jacques E.

2013-01-01

308

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

PubMed Central

?-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

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

309

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

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…

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

2006-01-01

310

Serum Amylase in Bulimia Nervosa and Purging Disorder: Differentiating the Association with Binge Eating versus Purging Behavior  

PubMed Central

Objective Elevated serum amylase levels in bulimia nervosa (BN), associated with increased salivary gland size and self-induced vomiting in some patients, provide a possible marker of symptom severity. The goal of this study was to assess whether serum hyperamylasemia in BN is more closely associated with binge eating episodes involving consumption of large amounts of food or with purging behavior. Method Participants included women with BN (n=26); women with “purging disorder” (PD), a subtype of EDNOS characterized by recurrent purging in the absence of objectively large binge eating episodes (n=14); and healthy non-eating disorder female controls (n=32). There were no significant differences in age or body mass index (BMI) across groups. The clinical groups reported similar frequency of self-induced vomiting behavior and were free of psychotropic medications. Serum samples were obtained after overnight fast and were assayed for alpha-amylase by enzymatic method. Results Serum amylase levels were significantly elevated in BN (60.7 ± 25.4 international units [IU]/liter, mean ± sd) in comparison to PD (44.7 ± 17.1 IU/L, p < 02) and to Controls (49.3 ± 15.8, p < .05). Conclusion These findings provide evidence to suggest that it is recurrent binge eating involving large amounts of food, rather than self-induced vomiting, which contributes to elevated serum amylase values in BN. PMID:21781981

Wolfe, Barbara E.; Jimerson, David C.; Smith, Adrian; Keel, Pamela K.

2011-01-01

311

Identification of alpha amylase inhibitors from Syzygium cumini Linn seeds.  

PubMed

The aqueous extract of S. cumini or Eugenia jambolana seeds and Psidium guajava leaves showed higher inhibition against the porcine pancreatic alpha-amylase among the medicinal plants studied. The alpha-amylase inhibitors from S. cumini seeds were separated from the extract by preparative thin layer chromatography into fractions with different Rf values. The fraction with Rf value between 0.285 and 0.43, which showed maximum inhibitory activity, was eluted and analyzed through LC-MS. The compounds identified from the seed extract ofS. cumini were betulinic acid and 3,5,7,4'-tetrahydroxy flavanone, which were reported earlier from S. formosanum and other plants. Dixon plot showed that the inhibition was noncompetitive in nature. PMID:18949899

Karthic, K; Kirthiram, K S; Sadasivam, S; Thayumanavan, B

2008-09-01

312

Purification and characterization of the extracellular. alpha. -amylase from Clostridium acetobutylicum ATCC 824  

SciTech Connect

The extracellular {alpha}-amylase (1,4-{alpha}-D-glucanglucanohydrolase; EC 3.2.1.1) from Clostridium acetobutylicum ATCC 824 was purified to homogeneity by anion-exchange chromatography (Mono Q) and gel filtration (Superose 12). The enzyme had an isoelectric point of 4.7 and a molecular weight of 84,000, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was a monomeric protein, the 19-amino-acid N terminus of which displayed 42% homology with the Bacillus subtilis saccharifying {alpha}-amylase. The amino acid composition of the enzyme showed a high number of acidic and hydrophobic residues and only one cysteine residue per mole. The activity of the {alpha}-amylase was not stimulated by calcium ions (or other metal ions) or inhibited by EDTA, although the enzyme contained seven calcium atoms per molecule. {alpha}-Amylase activity on soluble starch was optimal at pH 5.6 and 45{degree}C. The {alpha}-amylase was stable at an acidic pH but very sensitive to thermal inactivation. It hydrolyzed soluble starch, with a K{sub m} of 3.6 g {center dot} liter{sup {minus}1} and a K{sub cat} of 122 mol of reducing sugars {center dot} s{sup {minus}1} {center dot} mol{sup {minus}1}. The {alpha}-amylase showed greater activity with high-molecular-weight substrates than with low-molecular-weight maltooligosaccharides, hydrolyzed glycogen and pullulan slowly, but did not hydrolyze dextran or cyclodextrins. The major end products of maltohexaose degradation were glucose, maltose, and maltotriose; maltotetraose and maltopentaose were formed as intermediate products. Twenty seven percent of the glucoamylase activity generally detected in the culture supernatant of C. acetobutylicum can be attributed to the {alpha}-amylase.

Paquet, V.; Croux, C.; Goma, G.; Soucaille, P. (Centre National de la Recherche Scientifique Unite Associee, Toulouse (France))

1991-01-01

313

Purification and characterization of the extracellular alpha-amylase from Clostridium acetobutylicum ATCC 824.  

PubMed Central

The extracellular alpha-amylase (1,4-alpha-D-glucanglucanohydrolase; EC 3.2.1.1) from Clostridium acetobutylicum ATCC 824 was purified to homogeneity by anion-exchange chromatography (mono Q) and gel filtration (Superose 12). The enzyme had an isoelectric point of 4.7 and a molecular weight of 84,000, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was a monomeric protein, the 19-amino-acid N terminus of which displayed 42% homology with the Bacillus subtilis saccharifying alpha-amylase. The amino acid composition of the enzyme showed a high number of acidic and hydrophobic residues and only one cysteine residue per mole. The activity of the alpha-amylase was not stimulated by calcium ions (or other metal ions) or inhibited by EDTA, although the enzyme contained seven calcium atoms per molecule. alpha-Amylase activity on soluble starch was optimal at pH 5.6 and 45 degrees C. The alpha-amylase was stable at an acidic pH but very sensitive to thermal inactivation. It hydrolyzed soluble starch, with a Km of 3.6 g . liter-1 and a Kcat of 122 mol of reducing sugars . s-1 . mol-1. The alpha-amylase showed greater activity with high-molecular-weight substrates than with low-molecular-weight maltooligosaccharides, hydrolyzed glycogen and pullulan slowly, but did not hydrolyze dextran or cyclodextrins. The major end products of maltohexaose degradation were glucose, maltose, and maltotriose; maltotetraose and maltopentaose were formed as intermediate products. Twenty seven percent of the glucoamylase activity generally detected in the culture supernatant of C. acetobutylicum can be attributed to the alpha-amylase. Images PMID:8967771

Paquet, V; Croux, C; Goma, G; Soucaille, P

1991-01-01

314

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

PubMed

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

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

2014-04-01

315

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

PubMed Central

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

Goggin, Danica E.; Powles, Stephen B.

2012-01-01

316

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

PubMed Central

?-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

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

2014-01-01

317

Purification and characterization of midgut ?-amylase in a predatory bug, Andralus spinidens.  

PubMed

?-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

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

2014-01-01

318

Radiation-Induced Salivary Gland Dysfunction Results From p53-Dependent Apoptosis  

SciTech Connect

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.

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

319

Purification, characterization, and synergistic action of phytate-resistant ?-amylase and ?-glucosidase from Geobacillus thermodenitrificans HRO10  

Microsoft Academic Search

The ?-amylase (1, 4-?-d-glucanohydrolase; EC 3.2.1.1) and ?-glucosidase (?-d-glucoside glucohydrolase; EC 3.2.1.20) secreted by Geobacillus thermodenitrificans HRO10 were purified to homogeneity (13.6-fold; 11.5% yield and 25.4-fold; 32.0% yield, respectively) through a series of steps. The molecular weight of ?-amylase was 58kDa, as estimated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The ?-amylase activity on potato starch was optimal at pH

Thaddeus C. Ezeji; Hubert Bahl

2006-01-01

320

Nucleotide sequence of the beta-cyclodextrin glucanotransferase gene of alkalophilic Bacillus sp. strain 1011 and similarity of its amino acid sequence to those of alpha-amylases.  

PubMed Central

The nucleotide sequence of the gene for cyclodextrin glucanotransferase of alkalophilic Bacillus sp. strain 1011 was determined. The deduced amino acid sequence at the NH2-terminal side of the enzyme showed a high homology with the sequences of alpha-amylase in the three regions which constitutes the active centers of alpha-amylases. PMID:2957361

Kimura, K; Kataoka, S; Ishii, Y; Takano, T; Yamane, K

1987-01-01

321

Follicular lymphoma of the submandibular salivary gland  

PubMed Central

Lymphomas are neoplastic diseases of lymph nodes. Lymphoma of the salivary gland is rare accounting for less than 5% of lymphomas overall. Furthermore, lymphomas arising in the submandibular gland are reported to comprise 916% of all salivary gland lymphomas. Among lymphomas originating from salivary glands, the ratio of follicular lymphoma is very low. They can also be seen in the lymph nodes of the salivary glands which is an uncommon presentation. Here, we present a case follicular lymphoma which presented as a salivary gland tumour. PMID:25364171

Shashidara, R.; Prasad, Priyanka R.; Jaishankar; Joseph, Thomas

2014-01-01

322

Role of the lateral hypothalamus in submandibular salivary secretion during feeding in rats.  

PubMed

To evaluate the role of the lateral hypothalamic area (LH) in the masticatory-salivary reflex, we investigated submandibular salivary secretion and the electromyographic (EMG) activity of the jaw-closer masseter muscle in sham-operated rats and rats with unilateral LH lesions. One week prior to surgery and recording, the rats were given daily experience of eating pellets; powder; or hard, medium or soft mash, all of which were composed of laboratory chow. Salivary secretion was induced during eating and grooming behavior. During eating, the powdered food induced the highest salivary flow rate, and the soft (wet) mash induced the lowest salivary flow rate. Conversely, the amount of food consumed (dry weight) was greatest when soft mash was provided and lowest when the powder or pellets (a dry diet) were provided. The EMG activity of the masseter muscle during eating was greatest during consumption of the pellets and weakest during consumption of the powder. LH lesions that were ipsilateral to the examined submandibular gland reduced salivary secretion to about 20-30% of the control value, whereas contralateral LH lesions reduced it to about 40-50% of the control value. Neither masseter muscle EMG activity nor food consumption was markedly affected by the presence of an LH lesion. These results suggest that the texture of food, especially its water content, affects the flow rate of saliva and that the LH is heavily involved in the masticatory-salivary reflex. PMID:25446459

Matsuo, Ryuji; Kobashi, Motoi; Mitoh, Yoshihiro; Fujita, Masako

2015-01-30

323

Improvement of thermal stability of a mutagenised ?-amylase by manipulation of the calcium-binding site.  

PubMed

Site-directed mutagenesis of an ?-amylase isolated from Bacillus megaterium WHO has been performed to evaluate the roles of the calcium binding site residues in enzyme thermostability. The strategy used was to replace residues in the hypothetical calcium binding loops of B. megaterium WHO ?-amylase (BMW-amylase) by equivalent positions at Halothermothrix orenii ?-amylase (AmyA) as a thermophilic amylase by QuikChange site directed mutagenesis. Asn-75, Ser-76, and His-77 were mutated in the second calcium binding site which resulted in an increase in thermostability. All mutants retained their hydrolytic activity although their kcat parameter decreased in compare to the wild type and in the presence of calcium ions. In S76P and H77E, the Km for starch decreases while overall activity (kcat/Km) was increased. In the presence of calcium, conversion of His-77 to Glu resulted in a 4-fold enhancement in enzyme half life and a 9°C upward shift in T50, which was observed in compare to the wild type. Further analysis suggested the H77E mutant as the most stable which increased the affinity of the enzyme for calcium ion and its optimum temperature was 5°C higher than the wild type. PMID:24315644

Ghollasi, Marzieh; Ghanbari-Safari, Maryam; Khajeh, Khosro

2013-12-10

324

Production and biochemical characterization of a high maltotetraose (G4) producing amylase from Pseudomonas stutzeri AS22.  

PubMed

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 Ca(2+)-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

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

2014-01-01

325

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

PubMed Central

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

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

2014-01-01

326

Bioassay-guided separation of an alpha-amylase inhibitor anthocyanin from Vaccinium arctostaphylos berries.  

PubMed

Vaccinium arctostaphylos is a traditional medicinal plant in Iran used for the treatment of diabetes mellitus. In our search for antidiabetic compounds from natural sources, we found that the extract obtained from V. arctostaphylos berries showed an inhibitory effect on pancreatic alpha-amylase in vitro [IC50 = 1.91 (1.89-1.94) mg/mL]. The activity-guided purification of the extract led to the isolation of malvidin-3-O-beta-glucoside as an a-amylase inhibitor. The compound demonstrated a dose-dependent enzyme inihibitory activity [IC50 = 0.329 (0.316-0.342) mM]. PMID:21138057

Nickavar, Bahman; Amin, Gholamreza

2010-01-01

327

Purification and Characterization of a Hyperthermostable and High Maltogenic ?-Amylase of an Extreme Thermophile Geobacillus thermoleovorans  

Microsoft Academic Search

The purified ?-amylase of Geobacillus thermoleovorans had a molecular mass of 26 kDa with a pI of 5.4, and it was optimally active at 100 °C and pH 8.0. The T\\u000a 1\\/2 of ?-amylase at 100 °C increased from 3.6 to 5.6 h in the presence of cholic acid. The activation energy and temperature\\u000a quotient (Q\\u000a 10) of the enzyme were 84.10 kJ\\/mol and 1.31, respectively.

J. L. Uma Maheswar Rao; T. Satyanarayana

2007-01-01

328

Raw Starch Degrading Amylase Production by Various Fungal Cultures Grown on Cassava Waste  

PubMed Central

The solid waste of sago industry using cassava was fermented by Aspergillus niger, Aspergillus terreus and Rhizopus stolonifer in solid state fermentation. Cassava waste contained 52 per cent starch and 2.9 per cent protein by dry weight. The amylase activity was maintained at a high level and the highest amylase activity was observed on the 8th day in R. stolonifer mediated fermentation. R. stolonifer was more efficient than Aspergillus niger and Aspergillus terreus in bioconverting cassava waste into fungal protein (90.24 mg/g) by saccharifying 70% starch and releasing 44.5% reducing sugars in eight days of solid state fermentation. PMID:24039485

Balaji, P.; Eyini, M.

2006-01-01

329

Bioactivity-Guided Separation of an ?-Amylase Inhibitor Flavonoid from Salvia virgata  

PubMed Central

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

Nickavar, Bahman; Abolhasani, Leyla

2013-01-01

330

Alpha-amylase and alpha-glucosidase inhibition is differentially modulated by fucoidan obtained from Fucus vesiculosus and Ascophyllum nodosum.  

PubMed

Fucoidan is a water-soluble, negatively charged, biologically active polysaccharide found in great abundance in brown marine algae. However, the inhibition of ?-amylase and ?-glucosidase by fucoidan derived from two algal species (Ascophyllum nodosum and Fucus vesiculosus) harvested at different periods (accounting for seasonal and yearly variations) has never been investigated. It was found that fucoidans inhibited ?-glucosidase differently, depending on the algal species from which it was extracted and the algae's season of harvest. Fucoidan extracted from A. nodosum was a more potent inhibitor of ?-glucosidase, with an IC50 ranging from 0.013 to 0.047 mg/mL, than the inhibition by fucoidan extracted from F. vesiculosus (IC50=0.049 mg/mL). In contrast, fucoidan extracted from F. vesiculosus did not inhibit ?-amylase activity, while fucoidan from A. nodosum decreased ?-amylase activity by 7-100% at 5 mg/mL depending upon the algae harvest period. An IC50 of 0.12-4.64 mg/mL for fucoidan from A. nodosum was found for the ?-amylase inhibition. The ability of fucoidan to inhibit ?-amylase and ?-glucosidase thus varies according to the algae species and harvest period. A. nodosum is more suitable than F. vesiculosus as a source of fucoidan to inhibit ?-amylase and ?-glucosidase activities. Their potential benefits towards Type 2 diabetes management should be further investigated. PMID:24388677

Kim, Kyung-Tae; Rioux, Laurie-Eve; Turgeon, Sylvie L

2014-02-01

331

Salivary cortisol levels are associated with outcomes of weight reduction therapy in obese Japanese patients.  

PubMed

Dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis can increase the risk of cardiovascular disease (CVD). However, the detailed relationships of HPA axis activity with weight reduction and CVD risk factors in obese patients have not been examined. This study was designed to elucidate the associations of salivary cortisol levels with weight reduction and CVD risk factors in obese patients. As a marker of HPA axis activity, we measured the morning salivary cortisol levels of 83 obese Japanese outpatients. We also examined metabolic parameters, inflammatory markers, and indicators of arterial stiffness, that is, the pulse wave velocity and cardio-ankle vascular index. All 83 obese patients underwent 3-month weight reduction therapy with lifestyle modification. At the baseline, multivariate regression analysis revealed that only logarithmic transformation of C-reactive protein (? = 0.258, P < .05) and cardio-ankle vascular index (? = 0.233, P < .05) were independent determinants of the salivary cortisol levels. However, other metabolic parameters were not significantly associated with the salivary cortisol levels. In addition, lower salivary cortisol levels and higher body weight at the baseline were the only independent determinants of successful weight loss through the weight reduction therapy (P < .01). The present study demonstrates that the baseline morning salivary cortisol levels are significantly associated with the levels of an inflammatory marker, arterial stiffness, and successful weight reduction in obese patients. Therefore, salivary cortisol could be a useful marker for assessing and managing body weight and CVD risk factors in obese patients. PMID:21871641

Himeno, Akihiro; Satoh-Asahara, Noriko; Usui, Takeshi; Wada, Hiromichi; Tochiya, Mayu; Kono, Shigeo; Yamada-Goto, Nobuko; Katsuura, Goro; Hasegawa, Koji; Nakao, Kazuwa; Shimatsu, Akira

2012-02-01

332

Occurrence of toxicity among protease, amylase, and color mutants of a nontoxic soy sauce koji mold  

SciTech Connect

A soy sauce koji mold, Aspergillus flavus var. columnaris Raper and Fennel (ATCC 44310), was treated with UV irradiation to obtain mutant strains possessing high protease activities, high amylase activities, and light-colored conidia. Selected mutant strains were tested for toxicity, and some were found acutely toxic to weanling rats, although all were negative for aflatoxin production.

Kalayanamitr, A.; Bhumiratana, A.; Flegel, T.W.; Glinsukon, T.; Shinmyo, A.

1987-08-01

333

Occurrence of toxicity among protease, amylase, and color mutants of a nontoxic soy sauce koji mold.  

PubMed Central

A soy sauce koji mold, Aspergillus flavus var. columnaris Raper and Fennel (ATCC 44310), was treated with UV irradiation to obtain mutant strains possessing high protease activities, high amylase activities, and light-colored conidia. Selected mutant strains were tested for toxicity, and some were found acutely toxic to weanling rats, although all were negative for aflatoxin production. PMID:2444160

Kalayanamitr, A; Bhumiratana, A; Flegel, T W; Glinsukon, T; Shinmyo, A

1987-01-01

334

PURIFICATION AND CLONING OF THE SALIVARY PEROXIDASE\\/CATECHOL OXIDASE OF THE MOSQUITO ANOPHELES ALBIMANUS  

Microsoft Academic Search

Salivary homogenates of the adult female mosquito Anopheles albimanus have been shown previously to contain a vasodilatory activity associated with a catechol oxidase\\/peroxidase activity. We have now purified the salivary peroxidase using high-performance liquid chromatography. The pure enzyme is able to relax rabbit aortic rings pre-constricted with norepinephrine. The peroxidase has a relative molecular mass of 66 907 as estimated

JOSÉ M. C. RIBEIRO; JESUS G. VALENZUELA

335

Regulation of the amylase secretion by the yeast Filobasidium capsuligenum and a 2-deoxy-d-glucose resistant mutant  

Microsoft Academic Search

The regulation of extracellular amylase production by the basidiomycetous yeast Filobasidium capsuligenum CCY 64-5-1 was characterized using growing and resting cells. A basal level of amylolytic activity was produced with various carbon sources including glucose. Amylase secretion was repressed by glucose and, more severely, by 2-deoxy-d-glucose, whereas compounds with a-1,4-linked glucose, such as methyl glucoside, maltose, ß-cyclodextrin and soluble starch,

René Mot; Hubert Verachtert

1987-01-01

336

Immobilization of a thermostable ?-amylase, Termamyl ®, onto nitrocellulose membrane by Cibacron Blue F3GA dye binding  

Microsoft Academic Search

Highly porous nitrocellulose membranes were prepared by a solvent casting technique for the first time to immobilize ?-amylase. An affinity dye, namely Cibacron Blue F3GA (CB), was incorporated covalently within the structure. The nitrocellulose–CB derivatized membranes were used for the immobilization of a starch degrading enzyme, ?-amylase. Optimum conditions of immobilization for highest apparent activity were determined as pH 6.0,

Deniz Tanyolaç; Belma I??k Yürüksoy; Ahmet R Özdural

1998-01-01

337

Halophilic alkali- and thermostable amylase from a novel polyextremophilic Amphibacillus sp. NM-Ra2.  

PubMed

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

Mesbah, Noha M; Wiegel, Juergen

2014-09-01

338

?-amylase from starchless seeds of Trigonella foenum-graecum and its localization in germinating seeds.  

PubMed

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

Srivastava, Garima; Kayastha, Arvind M

2014-01-01

339

CHARACTERIZATION OF BARLEY TISSUE-UBIQUITOUS B-AMYLASE2  

Technology Transfer Automated Retrieval System (TEKTRAN)

There are two barley b-amylases genes, encoding important starch degrading enzymes. The endosperm-specific b-amylase (Bmy1), the more abundant isozyme in cereal seeds, has been thoroughly characterized. The lesser abundant b-amylase2 (Bmy2), has not been biochemically characterized from any cereal s...

340

Cloning and Expression Analysis of the Bombyx mori ?-amylase Gene (Amy) from the Indigenous Thai Silkworm Strain, Nanglai  

PubMed Central

?-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

Ngernyuang, Nipaporn; Kobayashi, Isao; Promboon, Amornrat; Ratanapo, Sunanta; Tamura, Toshiki; Ngernsiri, Lertluk

2011-01-01

341

The role of secretory granules in radiation-induced dysfunction of rat salivary glands  

SciTech Connect

To investigate the possible role of secretory granules in radiation-induced salivary gland dysfunction, rats were pretreated with isoproterenol (5 mg/kg intraperitoneally) to degranulate salivary gland acini. At maximal depletion, salivary glands were locally irradiated with a single dose of 15 Gy of X rays. Parotid and submandibular/sublingual saliva samples were collected before and 1-10 days after irradiation. The lag phase, flow rate, concentrations of potassium and sodium, and amylase secretion were determined. Sham-treated, isoproterenol-treated and irradiated animals provided reference data. In the parotid gland, but not in the submandibular gland, protection against radiation-induced changes in flow rate and composition of saliva occurred after pretreatment with isoproterenol. Combining morphological data from a previous study with data from the current study, it is suggested that improvement of parotid gland function is attributed predominantly to a proliferative stimulus on acinar cells by isoproterenol and not to its degranulation effect. After pretreatment with isoproterenol, an earlier expression of radiation-induced acinar cell damage leading to death was observed, followed by a faster tissue recovery. Thus the proliferative stimulus on acinar cells may accelerate the unmasking of latent lethal damage, resulting in the earlier replacement of dead cells by new, functionally intact cells. 33 refs., 2 figs.

Peter, B.; Van Waarde, M.A.W.H.; Konings, A.W.T. [Univ. of Groningen (Netherlands); Vissink, A. [Univ. of Groningen (Netherlands)]|[Univ. Hospital, Groningen (Netherlands); `s-Gravenmade, E.J. [Univ. Hospital, Groningen (Netherlands)

1995-02-01

342

Spectral effect: each population must have its own normal midnight salivary cortisol reference values determined  

PubMed Central

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

Tanakol, Refik; Karpuzoglu, Hande; Abbasoglu, Semra; Yarman, Sema; Boztepe, Harika; Alagol, Faruk

2013-01-01

343

Properties and gene structure of the Thermotoga maritima alpha-amylase AmyA, a putative lipoprotein of a hyperthermophilic bacterium.  

PubMed Central

Thermotoga maritima MSB8 has a chromosomal alpha-amylase gene, designated amyA, that is predicted to code for a 553-amino-acid preprotein with significant amino acid sequence similarity to the 4-alpha-glucanotransferase of the same strain and to alpha-amylase primary structures of other organisms. Upstream of the amylase gene, a divergently oriented open reading frame which can be translated into a polypeptide with similarity to the maltose-binding protein MalE of Escherichia coli was found. The T. maritima alpha-amylase appears to be the first known example of a lipoprotein alpha-amylase. This is in agreement with observations pointing to the membrane localization of this enzyme in T. maritima. Following the signal peptide, a 25-residue putative linker sequence rich in serine and threonine was found. The amylase gene was expressed in E. coli, and the recombinant enzyme was purified and characterized. The molecular mass of the recombinant enzyme was estimated at 61 kDa by denaturing gel electrophoresis (63 kDa by gel permeation chromatography). In a 10-min assay at the optimum pH of 7.0, the optimum temperature of amylase activity was 85 to 90 degrees C. Like the alpha-amylases of many other organisms, the activity of the T. maritima alpha-amylase was dependent on Ca2+. The final products of hydrolysis of soluble starch and amylose were mainly glucose and maltose. The extraordinarily high specific activity of the T. maritima alpha-amylase (about 5.6 x 10(3) U/mg of protein at 80 degrees C, pH 7, with amylose as the substrate) together with its extreme thermal stability makes this enzyme an interesting candidate for biotechnological applications in the starch processing industry. PMID:9006052

Liebl, W; Stemplinger, I; Ruile, P

1997-01-01

344

Digestive amylase from the larger grain borer, Prostephanus truncatus Horn.  

PubMed

A combination of ion-exchange chromatography, preparative electrophoresis and gel filtration chromatography allowed a 1209-fold purification of one of the two major digestive alpha-amylases from larvae of the larger grain borer, Prostephanus truncatus Horn. The purified enzyme showed a molecular mass of 60.2 kDa, an isoelectric point of 4.7 and an optimal pH for activity of 6.0. The enzyme was heat labile and it was recognized by proteinaceous inhibitors from amaranth seeds (Amaranthus hypochondriacus), whereas extracts from maize (Zea mays) and tepary bean (Phaseolus acutifolius) produced very low inhibition. When the enzyme was measured at different stages of development, maximal activity was found in the second instar larvae. Activity drastically decreased to a very low level during the pupae stage and increased again at the adult stage. A zymogram of the different developmental stages showed two main bands of alpha-amylase activity, which almost disappeared at the pupae stage to increase again during the adult stage, revealing a new, smaller band. This new band may be required for a better adaptation of the adult insect to its new environment. PMID:11007185

Mendiola-Olaya, E; Valencia-Jiménez, A; Valdés-Rodríguez, S; Délano-Frier, J; Blanco-Labra, A

2000-07-01

345

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 ...

346

Local entropy difference upon a substrate binding of a psychrophilic ?-amylase and a mesophilic homologue  

NASA Astrophysics Data System (ADS)

Psychrophilic ?-amylase from the antarctic bacterium pseudoalteromonashaloplanktis (AHA) and its mesophilic homologue, porcine pancreatic ?-amylase (PPA) are theoretically investigated with molecular dynamics (MD) simulations. We carried out 240-ns MD simulations for four systems, AHA and PPA with/without the bound substrate, and examined protein conformational entropy changes upon the substrate binding. We developed an analysis that decomposes the entropy changes into contributions of individual amino acids, and successfully identified protein regions responsible for the entropy changes. The results provide a molecular insight into the structural flexibilities of those enzymes related to the temperature dependences of the enzymatic activity.

Kosugi, Takahiro; Hayashi, Shigehiko

2011-01-01

347

Study of fluorescence quenching of Barley ?-amylase  

NASA Astrophysics Data System (ADS)

The fluorescence quenching of Barley ?-amylase by acrylamide and succinimide has been studied in water using steady-state and time-resolved fluorescence techniques. The steady-state fluorescence quenching technique has been performed in three different pHs (i.e., 6, 7 and 8) of water. Ground state and excited state binding constants (Kg &Ke) have been calculated. From the calculated binding constants (Kg &Ke) the free energy changes for the ground (?Gg) and excited (?Ge) states have been calculated and are presented in tables. UV and FTIR spectra have also been recorded to prove the binding of Barley ?-amylase with acrylamide and succinimide.

Bakkialakshmi, S.; Shanthi, B.; Bhuvanapriya, T.

2012-05-01

348

Comparisons of salivary proteins from five aphid (Hemiptera: Aphididae) species.  

PubMed

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

Cooper, W Rodney; Dillwith, Jack W; Puterka, Gary J

2011-02-01

349

Structural stability of soybean (Glycine max) ?-amylase: properties of the unfolding transition studied with fluorescence and CD spectroscopy.  

PubMed

Stability and unfolding of mammalian and microbial ?-amylases have been intensively investigated. However, there is only limited information available on the structural stability of plant ?-amylases, namely of the two isoenzymes from barley AMY1 and AMY2, of the ?-amylase from mung bean (Vigna radiata), and of the ?-amylase from malted sorghum (Sorghum bicolor). We report here the stability of soyabean ?-amylase (GMA), against elevated temperatures and chemical denaturants (GndHCl) by employing circular dichroism and fluorescence spectroscopy. Since it is well-known that calcium ions play a crucial role for enzymatic activity and stability of a-amylases, we performed our studies with calcium bound and calcium free GMA. The thermal unfolding transition temperature decreased from 72°C for calcium saturated samples to 57°C for the case of calcium depleted GMA. Similarly, the GndHCl transition concentration was lowered from 0.70 M for calcium bound GMA to 0.41 M in the absence of calcium. Thermal unfolding of GMA irreversible due to aggregation of the unfolded state. GMA unfolded in 6 M GndHCl shows high degree of reversibility after diluting the unfolded enzyme in native buffer containing 7 M glycerol. Furthermore, the refolded enzyme showed 93% of activity. PMID:20955173

Kumari, Arpana; Rosenkranz, Tobias; Fitter, Jörg; Kayastha, Arvind M

2011-03-01

350

Isolation and characterization of a proteinaceous ?-amylase inhibitor AAI-CC5 from Streptomyces sp. CC5, and its gene cloning and expression.  

PubMed

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

Sun, Zhibin; Lu, Weihao; Liu, Pingping; Wang, Hui; Huang, Yan; Zhao, Yuguo; Kong, Yi; Cui, Zhongli

2014-11-20

351

Carbonic anhydrase in minor salivary glands of quail: histochemistry versus immunohistochemistry.  

PubMed

Studies on the mechanisms of saliva secretion have indicated that carbonic anhydrase (CA) is expressed in mammalian salivary glands. The enzyme is present in the saliva as the only known secretory isoenzyme, CAVI; its activity has been related to the modulation of taste and caries development. Unlike mammals, in birds, saliva is produced by the so-called minor salivary glands, mostly concentrated in the tongue. The involvement of CA has never been explored in avian salivary secretion. Thus, we aimed here to ascertain the enzyme occurrence in the quail lingual glands by a parallel investigation of the distributional patterns of CA activity sites, as visualized by histochemistry, and the immunohistochemical patterns of cytosolic CAII and secretory CAVI. The comparative evaluation of our findings does not rule out that some CA isoforms, associated to basolateral borders of the secretory cells and antigenically different from cytosolic CAII and secretory CAVI, may be involved in the salivary secretion in the quail lingual glands. PMID:23323952

Gabrielli, Maria Gabriella; Tomassoni, Daniele

2014-02-01

352

Tactics for modeling multiple salivary analyte data in relation to behavior problems: Additive, ratio, and interaction effects.  

PubMed

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

Chen, Frances R; Raine, Adrian; Granger, Douglas A

2015-01-01

353

Adsorption of ?-amylase onto poly(acrylamide/maleic acid) hydrogels  

NASA Astrophysics Data System (ADS)

Poly(acrylamide/maleic acid) [P(AAm/MA)] hydrogels were prepared by irradiating the ternary mixtures of AAm/MA and water by ? rays at ambient temperature. The influence of the MA on the adsorption of ?-amylase, optimum working conditions and storage stability of enzyme were investigated. The adsorption capacity of hydrogels were found to increase from 0.40 to 0.71 mg ?-amylase/g dry gel with increasing amount of MA in the gel system. Maximum enzyme activities were observed at lower pH values and higher temperatures for adsorbed enzyme compared with free enzyme. Kinetic parameters were calculated as 2.51 g/dm 3 for Km and 1.67×10 -3 g/dm 3 min for Vmax for free enzyme and in the range of 12.3-12.9 g/dm 3 for Km and 1.63×10 -3-1.96×10 -3 g/dm 3 min for Vmax depending on the amount of MA in the hydrogel. While, the enzymatic activity of free enzyme was completely lost after 20 days, adsorbed enzyme retained 47-59% of its original activity after 20 days, depending on the amount of MA in the hydrogels.

Tümtürk, H.; Çaykara, T.; ?en, M.; Güven, O.

1999-08-01

354

Programmed cell death in the larval salivary glands of Apis mellifera (Hymenoptera, Apidae)  

Microsoft Academic Search

The morphological and histochemical features of degeneration in honeybee (Apis mellifera) salivary glands were investigated in 5th instar larvae and in the pre-pupal period. The distribution and activity patterns\\u000a of acid phosphatase enzyme were also analysed. As a routine, the larval salivary glands were fixed and processed for light\\u000a microscopy and transmission electron microscopy. Tissue sections were subsequently stained with

E C M Silva-Zacarin; G A Tomaino; M R Brocheto-Braga; S R Taboga; R L M Silva de Moraes

2007-01-01

355

Effect of negative air ions on computer operation, anxiety and salivary chromogranin Alike immunoreactivity  

Microsoft Academic Search

The effects of negative air ions on computer operation were examined using a biochemical index of the activity of the sympathetic\\/adrenomedullary system (i.e. salivary chromogranin A-like immunoreactivity (CgA-like IR)) and a self-report questionnaire (State-Trait Anxiety Inventory, Anxiety State—STAI-S). Twelve female students carried out a word processing task for 40 min. The salivary CgA-like IR increased more than three times on

Hideo Nakane; Osamu Asami; Yukio Yamada; Hideki Ohira

2002-01-01

356

Salivary Gland Development: A Template for Regeneration  

PubMed Central

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

Patel, Vaishali N.; Hoffman, Matthew P.

2014-01-01

357

Effects of high pressure on amylases and starch in wheat and barley flours  

Microsoft Academic Search

Application of high pressures (to 800 MPa) to 25% ww slurries of barley and wheat flours indicated that pressures above 300 MPa caused an increase in the action of the ?- and ?-amylases due to starch gelatinisation, the apparent activity increasing with pressure to a maximum at 500–600 MPa. At higher pressures marked losses in action were observed. Pressure treatment

Maria Regina A. Gomes; Richard Clark; Dave A. Ledward

1998-01-01

358

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...

359

Molecular cloning and characterization of amylase from soil metagenomic library derived from Northwestern Himalayas.  

PubMed

The increasing demand for novel biocatalysts stimulates exploration of resources from soil. Metagenomics, a culture independent approach, represent a sheer unlimited resource for discovery of novel biocatalysts from uncultured microorganisms. In this study, a soil-derived metagenomic library containing 90,700 recombinants was constructed and screened for lipase, cellulase, protease and amylase activity. A gene (pAMY) of 909 bp encoding for amylase was found after the screening of 35,000 Escherichia coli clones. Amino acid sequence comparison and phylogenetic analysis indicated that pAMY was closely related to uncultured bacteria. The molecular mass of pAMY was estimated about 38 kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Amylase activity was determined using soluble starch, amylose, glycogen and maltose as substrates. The maximal activity (2.46 U/mg) was observed at 40 degrees C under nearly neutral pH conditions with amylose; whereas it retains 90% of its activity at low temperature with all the substrates used in this study. The ability of pAMY to work at low temperature is unique for amylases reported so far from microbes of cultured and uncultured division. PMID:20054535

Sharma, Sarika; Khan, Farrah Gul; Qazi, Ghulam Nabi

2010-05-01

360

Modes of Inhibition of ?-Amylase and ?-Glucosidase by Aqueous Extract of Morinda lucida Benth Leaf  

PubMed Central

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

Kazeem, M. I.; Adamson, J. O.; Ogunwande, I. A.

2013-01-01

361

Relationship of sequence and structure to specificity in the ?-amylase family of enzymes  

Microsoft Academic Search

The hydrolases and transferases that constitute the ?-amylase family are multidomain proteins, but each has a catalytic domain in the form of a (?\\/?)8-barrel, with the active site being at the C-terminal end of the barrel ?-strands. Although the enzymes are believed to share the same catalytic acids and a common mechanism of action, they have been assigned to three

E. Ann MacGregor; Štefan Jane?ek; Birte Svensson

2001-01-01

362

Kinetic Analysis of Amylase Using Quantitative Benedict's and Iodine Starch Reagents  

ERIC Educational Resources Information Center

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…

Cochran, Beverly; Lunday, Deborah; Miskevich, Frank

2008-01-01

363

Replication of Oral BK Virus in Human Salivary Gland Cells  

PubMed Central

BK polyomavirus (BKPyV) is the most common viral pathogen among allograft patients. Increasing evidence links BKPyV to the human oral compartment and to HIV-associated salivary gland disease (HIVSGD). To date, few studies have analyzed orally derived BKPyV. This study aimed to characterize BKPyV isolated from throat wash (TW) samples from HIVSGD patients. The replication potential of HIVSGD-derived clinical isolates HIVSGD-1 and HIVSGD-2, both containing the noncoding control region (NCCR) architecture OPQPQQS, were assessed and compared to urine-derived virus. The BKPyV isolates displayed significant variation in replication potential. Whole-genome alignment of the two isolates revealed three nucleotide differences that were analyzed for a potential effect on the viral life cycle. Analysis revealed a negligible difference in NCCR promoter activity despite sequence variation and emphasized the importance of functional T antigen (Tag) for efficient replication. HIVSGD-1 encoded full-length Tag, underwent productive infection in both human salivary gland cells and kidney cells, and expressed viral DNA and Tag protein. Additionally, HIVSGD-1 generated DNase-resistant particles and by far surpassed the replication potential of the kidney-derived isolate in HSG cells. HIVSGD-2 encoded a truncated form of Tag and replicated much less efficiently. Quantitation of infectious virus, via the fluorescent forming unit assay, suggested that HIVSGD BKPyV had preferential tropism for salivary gland cells over kidney cells. Similarly, the results suggested that kidney-derived virus had preferential tropism for kidney cells over salivary gland cells. Evidence of HIVSGD-derived BKPyV oral tropism and adept viral replication in human salivary gland cells corroborated the potential link between HIVSGD pathogenesis and BKPyV. PMID:24173219

Burger-Calderon, Raquel; Madden, Victoria; Hallett, Ryan A.; Gingerich, Aaron D.; Nickeleit, Volker

2014-01-01

364

Resolvin D1 prevents TNF-?-mediated disruption of salivary epithelial formation  

PubMed Central

Sjögren's syndrome is a chronic autoimmune disorder characterized by inflammation of salivary glands resulting in impaired secretory function. Our present studies indicate that chronic exposure of salivary epithelium to TNF-? and/or IFN-? alters tight junction integrity, leading to secretory dysfunction. Resolvins of the D-series (RvDs) are endogenous lipid mediators derived from DHA that regulate excessive inflammatory responses leading to resolution and tissue homeostasis. In this study, we addressed the hypothesis that activation of the RvD1 receptor ALX/FPR2 in salivary epithelium prevents and/or resolves the TNF-?-mediated disruption of acinar organization and enhances monolayer formation. Our results indicate that 1) the RvD1 receptor ALX/FPR2 is present in fresh, isolated cells from mouse salivary glands and in cell lines of salivary origin; and 2) the agonist RvD1 (100 ng/ml) abolished tight junction and cytoskeletal disruption caused by TNF-? and enhanced cell migration and polarity in salivary epithelium. These effects were blocked by the ALX/FPR2 antagonist butyloxycarbonyl-Phe-Leu-Phe-Leu-Phe. The ALX/FPR2 receptor signals via modulation of the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathways since, in our study, blocking PI3K activation with LY294002, a potent and selective PI3K inhibitor, prevented RvD1-induced cell migration. Furthermore, Akt gene silencing with the corresponding siRNA almost completely blocked the ability of Par-C10 cells to migrate. Our findings suggest that RvD1 receptor activation promotes resolution of inflammation and tissue repair in salivary epithelium, which may have relevance in the restoration of salivary gland dysfunction associated with Sjögren's syndrome. PMID:22237406

Odusanwo, Olutayo; Chinthamani, Sreedevi; McCall, Andrew; Duffey, Michael E.

2012-01-01

365

Influence of substratum hydrophobicity on salivary pellicles: organization or composition?  

PubMed

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

Aroonsang, Watcharapong; Sotres, Javier; El-Schich, Zahra; Arnebrant, Thomas; Lindh, Liselott

2014-10-01

366

Genetic Regulation of Tissue-Specific Expression of Amylase Structural Genes in Drosophila melanogaster  

Microsoft Academic Search

Laboratory strains of Drosophila melanogaster were screened for spatial variations in adult midgut alpha -amylase (1,4-alpha -D-glucan glucanohydrolase, EC 3.2.1.1) expression. No strain-specific differences were found anteriorly, but three patterns of activity were discerned in the posterior midgut: A, activity throughout most of the region; B, activity in the anterior part of the region; and C, little or no activity.

Irene Abraham; Winifred W. Doane

1978-01-01

367

Antidiabetic Indian plants: a good source of potent amylase inhibitors.  

PubMed

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

Bhat, Menakshi; Zinjarde, Smita S; Bhargava, Shobha Y; Kumar, Ameeta Ravi; Joshi, Bimba N

2011-01-01

368

Salivary duct carcinoma in the mandible.  

PubMed

We reported 1 case of salivary duct carcinoma (SDC) in the mandible. The patient complained of pain and a growing mass in the right submandibular area for approximately 2 months. On clinical examination, there was a mass under the right angle of the mandible with a size of approximately 3 × 3 cm, a smooth surface, a poor activity, and a hard texture. Panoramic radiograph revealed poorly circumscribed area. Computed tomography presented mandible central destruction. Biopsy examination showed a malignant tumor that originated in the central epithelium of the mandible. An operation of unilateral selective neck dissection and mandible subtotal ectomy was performed. Postoperative pathology reported SDC. The patient received postoperative radiation and stayed alive at last follow-up without disease recurrence. Ablative resection and postoperative radiotherapy were the standard treatment stratagem for SDC, but trastuzumab therapy might play a key role in treating the disease in future. PMID:25377985

Shi, Shuang; Fang, Qi-Gen; Sun, Changfu

2014-11-01

369

Molecular cloning of alpha-amylases from cotton boll weevil, Anthonomus grandis and structural relations to plant inhibitors: an approach to insect resistance.  

PubMed

Anthonomus grandis, the cotton boll weevil, causes severe cotton crop losses in North and South America. Here we demonstrate the presence of starch in the cotton pollen grains and young ovules that are the main A. grandis food source. We further demonstrate the presence of alpha-amylase activity, an essential enzyme of carbohydrate metabolism for many crop pests, in A. grandis midgut. Two alpha-amylase cDNAs from A. grandis larvae were isolated using RT-PCR followed by 5' and 3' RACE techniques. These encode proteins with predicted molecular masses of 50.8 and 52.7kDa, respectively, which share 58% amino acid identity. Expression of both genes is induced upon feeding and concentrated in the midgut of adult insects. Several alpha-amylase inhibitors from plants were assayed against A. grandis alpha-amylases but, unexpectedly, only the BIII inhibitor from rye kernels proved highly effective, with inhibitors generally active against other insect amylases lacking effect. Structural modeling of Amylag1 and Amylag2 showed that different factors seem to be responsible for the lack of effect of 0.19 and alpha-AI1 inhibitors on A. grandis alpha-amylase activity. This work suggests that genetic engineering of cotton to express alpha-amylase inhibitors may offer a novel route to A. grandis resistance. PMID:12744224

Oliveira-Neto, Osmundo B; Batista, João A N; Rigden, Daniel J; Franco, Octávio L; Falcão, Rosana; Fragoso, Rodrigo R; Mello, Luciane V; dos Santos, Roseane C; Grossi-de-Sá, Maria F

2003-01-01

370

Purification and characterization of the extracellular alpha-amylase from Streptococcus bovis JB1.  

PubMed

The extracellular alpha-amylase (1,4-alpha-D-glucanglucanohydrolase; EC 3.2.1.1) from maltose-grown Streptococcus bovis JB1 was purified to apparent homogeneity by ion-exchange chromatography (Mono Q). The enzyme had an isoelectric point of 4.50 and an apparent molecular mass of 77,000 Da, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme was rich in acidic and hydrophobic amino acids. The 15-amino-acid NH2-terminal sequence was 40% homologous with the Bacillus subtilis saccharifying alpha-amylase and 27% homologous with the Clostridium acetobutylicum alpha-amylase. alpha-Amylase activity on soluble starch was optimal at pH 5.0 to 6.0. The enzyme was relatively stable between pH 5.5 and 8.5 and at temperatures below 50 degrees C. When soluble potato starch was used as the substrate, the enzyme had a Km of 0.88 mg.ml-1 and a kcat of 2,510 mumol of reducing sugar.min-1.mg of protein-1. The enzyme exhibited neither pullulanase nor dextranase activity and was 40 to 70% as active on amylopectin as on amylose. The major end products of amylose hydrolysis were maltose, maltotriose, and maltotetraose. PMID:8517735

Freer, S N

1993-05-01

371

Purification and characterization of the extracellular alpha-amylase from Streptococcus bovis JB1.  

PubMed Central

The extracellular alpha-amylase (1,4-alpha-D-glucanglucanohydrolase; EC 3.2.1.1) from maltose-grown Streptococcus bovis JB1 was purified to apparent homogeneity by ion-exchange chromatography (Mono Q). The enzyme had an isoelectric point of 4.50 and an apparent molecular mass of 77,000 Da, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme was rich in acidic and hydrophobic amino acids. The 15-amino-acid NH2-terminal sequence was 40% homologous with the Bacillus subtilis saccharifying alpha-amylase and 27% homologous with the Clostridium acetobutylicum alpha-amylase. alpha-Amylase activity on soluble starch was optimal at pH 5.0 to 6.0. The enzyme was relatively stable between pH 5.5 and 8.5 and at temperatures below 50 degrees C. When soluble potato starch was used as the substrate, the enzyme had a Km of 0.88 mg.ml-1 and a kcat of 2,510 mumol of reducing sugar.min-1.mg of protein-1. The enzyme exhibited neither pullulanase nor dextranase activity and was 40 to 70% as active on amylopectin as on amylose. The major end products of amylose hydrolysis were maltose, maltotriose, and maltotetraose. Images PMID:8517735

Freer, S N

1993-01-01

372

Lapatinib in Treating Patients With Recurrent and/or Metastatic Adenoid Cystic Cancer or Other Salivary Gland Cancers  

ClinicalTrials.gov

High-grade Salivary Gland Carcinoma; High-grade Salivary Gland Mucoepidermoid Carcinoma; Low-grade Salivary Gland Carcinoma; Low-grade Salivary Gland Mucoepidermoid Carcinoma; Recurrent Adenoid Cystic Carcinoma of the Oral Cavity; Recurrent Salivary Gland Cancer; Salivary Gland Acinic Cell Tumor; Salivary Gland Adenocarcinoma; Salivary Gland Adenoid Cystic Carcinoma; Salivary Gland Malignant Mixed Cell Type Tumor

2013-10-10

373

Purification, Characterization, and Potential of Saline Waste Water Remediation of a Polyextremophilic ?-Amylase from an Obligate Halophilic Aspergillus gracilis  

PubMed Central

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

Ali, Imran; Akbar, Ali; Yanwisetpakdee, Benjawan; Prasongsuk, Sehanat; Lotrakul, Pongtharin; Punnapayak, Hunsa

2014-01-01

374

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

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

Ronald D Mathison; Joseph S Davison; A Dean Befus; Daniel A Gingerich

2010-01-01

375

Morphological aspects of Culex quinquefasciatus salivary glands  

Microsoft Academic Search

The salivary glands of Culex quinquefasciatus female mosquitoes are paired organs composed of two lateral lobes with proximal and distal secretory portions, and a medial lobe. All portions comprise a simple epithelium that surrounds a salivary duct. In the apical portion of the medial lobe, non-secretory cells strongly resemble cells involved in ion and water transport.The general architecture of the

Tiago da Cunha Sais; Rosa Maria de Moraes; Paulo E. Ribolla; Antonio G. de Bianchi; Osvaldo Marinotti; A. Tania Bijovsky

2003-01-01

376

Distribution of lactoferrin in human salivary glands  

Microsoft Academic Search

An immunoperoxidase technique was used to study the distribution of lactoferrin (LF) in human salivary glands from autopsy tissues fixed in Carnoy's fluid for the optimal preservation of LF antigenicity. Specific LF staining was seen in the intralobular ducts of all salivary glands but never in the interlobular ducts; acinar LF was detected in most serous demilunes of the mixed

S. Reitamo; Y. T. Konttinen; M. Segerberg-Konttinen

1980-01-01

377

Extra EF hand unit (DX) mediated stabilization and calcium independency of ?-amylase.  

PubMed

It is the common feature of ?-amylases that calcium ion is required for their structural integrity and thermal stability. All amylases have at least one Ca(2+) per molecule; therefore amino acids involved in calcium binding are specific and conserved. In this study, sequence analysis revealed the presence of EF-hand-like motif in calcium-binding loop of Bacillus megaterium WHO (BMW)-amylase that was previously isolated from BMW. The EF-hand motif and its variants (EF-hand-like motif) are the most common calcium-binding motifs found in a large number of protein families. To investigate the effect of calcium ion on the thermal stability and activity of BMW-amylase, we used site-directed mutagenesis to replace histidine 58 with Asp (D), Ile (I), Tyr (Y), Phe (F), and Arg (R) at the seventh position of EF-hand-like motif. Upon the addition of an extra DX unit to the calcium-binding loop in H58D variant, thermal stability, catalytic activity, and chelating power of the enzyme improved due to higher affinity toward calcium. H58D variant demonstrated calcium independency compared to the wild type and other created mutants. Conformational changes in the presence and absence of Ca(2+) were monitored using fluorescence technique. PMID:22407721

Sadeghi, Leila; Khajeh, Khosro; Mollania, Nasrin; Dabirmanesh, Bahareh; Ranjbar, Bijan

2013-03-01

378

Iron binding modulates candidacidal properties of salivary histatin 5.  

PubMed

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

Puri, S; Li, R; Ruszaj, D; Tati, S; Edgerton, M

2015-01-01

379

Ornithodoros brasiliensis (mouro tick) salivary gland homogenates inhibit in vivo wound healing and in vitro endothelial cell proliferation.  

PubMed

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

Reck, José; Marks, Fernanda S; Termignoni, Carlos; Guimarães, Jorge A; Martins, João Ricardo

2013-04-01

380

Glycoinositolphospholipids from Trypanosomatids Subvert Nitric Oxide Production in Rhodnius prolixus Salivary Glands  

PubMed Central

Background Rhodnius prolixus is a blood-sucking bug vector of Trypanosoma cruzi and T. rangeli. T. cruzi is transmitted by vector feces deposited close to the wound produced by insect mouthparts, whereas T. rangeli invades salivary glands and is inoculated into the host skin. Bug saliva contains a set of nitric oxide-binding proteins, called nitrophorins, which deliver NO to host vessels and ensure vasodilation and blood feeding. NO is generated by nitric oxide synthases (NOS) present in the epithelium of bug salivary glands. Thus, T. rangeli is in close contact with NO while in the salivary glands. Methodology/Principal Findings Here we show by immunohistochemical, biochemical and molecular techniques that inositolphosphate-containing glycolipids from trypanosomatids downregulate NO synthesis in the salivary glands of R. prolixus. Injecting insects with T. rangeli-derived glycoinositolphospholipids (Tr GIPL) or T. cruzi-derived glycoinositolphospholipids (Tc GIPL) specifically decreased NO production. Salivary gland treatment with Tc GIPL blocks NO production without greatly affecting NOS mRNA levels. NOS protein is virtually absent from either Tr GIPL- or Tc GIPL-treated salivary glands. Evaluation of NO synthesis by using a fluorescent NO probe showed that T. rangeli-infected or Tc GIPL-treated glands do not show extensive labeling. The same effect is readily obtained by treatment of salivary glands with the classical protein tyrosine phosphatase (PTP) inhibitor, sodium orthovanadate (SO). This suggests that parasite GIPLs induce the inhibition of a salivary gland PTP. GIPLs specifically suppressed NO production and did not affect other anti-hemostatic properties of saliva, such as the anti-clotting and anti-platelet activities. Conclusions/Significance Taken together, these data suggest that trypanosomatids have overcome NO generation using their surface GIPLs. Therefore, these molecules ensure parasite survival and may ultimately enhance parasite transmission. PMID:23077586

Gazos-Lopes, Felipe; Mesquita, Rafael Dias; Silva-Cardoso, Lívia; Senna, Raquel; Silveira, Alan Barbosa; Jablonka, Willy; Cudischevitch, Cecília Oliveira; Carneiro, Alan Brito; Machado, Ednildo Alcantara; Lima, Luize G.; Monteiro, Robson Queiroz; Nussenzveig, Roberto Henrique; Folly, Evelize; Romeiro, Alexandre; Vanbeselaere, Jorick; Mendonça-Previato, Lucia; Previato, José Osvaldo; Valenzuela, Jesus G.; Ribeiro, José Marcos Chaves; Atella, Georgia Correa; Silva-Neto, Mário Alberto Cardoso

2012-01-01

381

Characterization of glucoamylase and ?-amylase from Monascus anka: enhanced production of ?-amylase in red koji.  

PubMed

To enhance glucoamylase and ?-amylase production from Monascus anka, we investigated the influence of different culture conditions on enzyme production and purified and characterized these enzymes. The effect of different raw materials was investigated by using solid-state plates of raw materials such as barley and non-waxy or waxy rice. The barley plate was the most suitable for mycelial growth, but glucoamylase and ?-amylase production per growth area did not differ according to the raw material. Investigation of the effect of temperature showed that incubation at 37 °C promoted maximal cell growth, while incubation at 25 °C and at 40 °C resulted in enhanced ?-amylase and glucoamylase production, respectively. Characterization of the purified enzymes revealed that ?-amylase was unstable at acidic pH and less resistant to heat (stable at < 40 °C) than glucoamylase. When these culture conditions were applied to enzyme production in red koji, reducing the temperature from 35 °C to 25 °C for 48 h in the late stages of growth resulted in higher glucoamylase and ?-amylase production (1.4 and 18 times, respectively) with a concomitant increase in protein stability. PMID:20708432

Yoshizaki, Yumiko; Susuki, Tomoka; Takamine, Kazunori; Tamaki, Hisanori; Ito, Kiyoshi; Sameshima, Yoshihiro

2010-12-01

382

Solid State Fermentation of a Raw Starch Digesting Alkaline Alpha-Amylase from Bacillus licheniformis RT7PE1 and Its Characteristics  

PubMed Central

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

Tabassum, Romana; Khaliq, Shazia; Rajoka, Muhammad Ibrahim; Agblevor, Foster

2014-01-01

383

SOURCES OF LARVAL SALIVARY GLAND SECRETION IN THE DIPTERAN CHIRONOMUS TENTANS  

PubMed Central

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

Doyle, D.; Laufer, H.

1969-01-01

384

Microencapsulation of purified amylase enzyme from pitaya (Hylocereus polyrhizus) peel in Arabic gum-chitosan using freeze drying.  

PubMed

Amylase is one of the most important enzymes in the world due to its wide application in various industries and biotechnological processes. In this study, amylase enzyme from Hylocereus polyrhizus was encapsulated for the first time in an Arabic gum-chitosan matrix using freeze drying. The encapsulated amylase retained complete biocatalytic activity and exhibited a shift in the optimum temperature and considerable increase in the pH and temperature stabilities compared to the free enzyme. Encapsulation of the enzyme protected the activity in the presence of ionic and non-ionic surfactants and oxidizing agents (H?O?) and enhanced the shelf life. The storage stability of amylase is found to markedly increase after immobilization and the freeze dried amylase exhibited maximum encapsulation efficiency value (96.2%) after the encapsulation process. Therefore, the present study demonstrated that the encapsulation of the enzyme in a coating agent using freeze drying is an efficient method to keep the enzyme active and stable until required in industry. PMID:24662085

Amid, Mehrnoush; Manap, Yazid; Zohdi, Nor Khanani

2014-01-01

385

Pupil and Salivary Indicators of Autonomic Dysfunction in Autism Spectrum Disorder  

PubMed Central

Dysregulated tonic pupil size has been reported in Autism Spectrum Disorder (ASD). Among the possible sources of this dysregulation are disruptions in the feedback loop between norepinephrine (NE) and hypothalamic systems. In the current study, we examined afternoon levels of salivary alpha-amylase (sAA, a putative correlate of NE) and cortisol (used to assess stress-based responses) in two independent samples of children with ASD. We found a larger pupil size and lower sAA levels in ASD, compared to typical and clinical age-matched controls. This was substantiated at the individual level, as sAA levels were strongly correlated with tonic pupil size. Relatively little diurnal variation in sAA taken in the home environment in the ASD group was also observed, while typical controls showed a significant linear increase throughout the day. Results are discussed in terms of potential early biomarkers and the elucidation of underlying neural dysfunction in ASD. PMID:22644965

Anderson, Christa J.; Colombo, John; Unruh, Kathryn E.

2013-01-01

386

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...Affirmed as GRAS § 184.1012 ?-Amylase enzyme preparation from Bacillus stearothermophilus. (a) ?-Amylase enzyme preparation is obtained...

2010-04-01

387

21 CFR 184.1012 - ?-Amylase enzyme preparation from Bacillus stearothermophilus.  

Code of Federal Regulations, 2012 CFR

...2012-04-01 2012-04-01 false α-Amylase enzyme preparation from Bacillus stearothermophilus...Affirmed as GRAS § 184.1012 ?-Amylase enzyme preparation from Bacillus stearothermophilus. (a) ?-Amylase enzyme preparation is obtained...

2012-04-01

388

21 CFR 184.1012 - ?-Amylase enzyme preparation from Bacillus stearothermophilus.  

...2014-04-01 2014-04-01 false α-Amylase enzyme preparation from Bacillus stearothermophilus...Affirmed as GRAS § 184.1012 ?-Amylase enzyme preparation from Bacillus stearothermophilus. (a) ?-Amylase enzyme preparation is obtained...

2014-04-01

389

21 CFR 184.1012 - ?-Amylase enzyme preparation from Bacillus stearothermophilus.  

Code of Federal Regulations, 2013 CFR

...2013-04-01 2013-04-01 false α-Amylase enzyme preparation from Bacillus stearothermophilus...Affirmed as GRAS § 184.1012 ?-Amylase enzyme preparation from Bacillus stearothermophilus. (a) ?-Amylase enzyme preparation is obtained...

2013-04-01

390

21 CFR 184.1012 - ?-Amylase enzyme preparation from Bacillus stearothermophilus.  

Code of Federal Regulations, 2011 CFR

...2011-04-01 2011-04-01 false α-Amylase enzyme preparation from Bacillus stearothermophilus...Affirmed as GRAS § 184.1012 ?-Amylase enzyme preparation from Bacillus stearothermophilus. (a) ?-Amylase enzyme preparation is obtained...

2011-04-01

391

Salivary alcohol dehydrogenase in non-smoking and smoking alcohol-dependent persons.  

PubMed

Increasing attention to the importance of saliva testing is not surprising because smoking and alcohol drinking act synergistically on oral tissues, and their metabolite levels, e.g., acetaldehyde, are much higher in saliva than in blood. The activity of salivary alcohol dehydrogenase (ADH) comes from oral microbiota, mucosa, and salivary glands. The purpose of this study was to investigate the involvement of ADH in the oral health pathology of smoking (AS) and non-smoking (ANS) alcohol-dependent males. The results indicated that the AS group had a more significant and longer duration (until the 30th day of alcohol abstinence) decrease in ADH activity and output than the ANS group (until the 15th day of alcohol abstinence) compared to controls (social drinkers; C). The decreased salivary flow (SF) in alcoholics was observed longer in the ANS group (until the 30th day of alcohol abstinence), whereas in the AS group SF normalized at the 15th day, probably due to the irritating effect of tobacco smoke on the oral mucosa. Because saliva was centrifuged to remove cells and debris (including microbial cells), the detected salivary ADH activity was derived from salivary glands and/or oral mucosa. A more profound and longer decrease in ADH activity/output in smoking than non-smoking alcoholics was likely due to the damaged salivary glands and/or oral mucosa, caused by the synergistic effect of alcohol drinking and smoking. The lower values of salivary ADH in smoking than non-smoking alcoholics might also be partly due to the reversed/inhibited ADH reaction by high levels of accumulated acetaldehyde. PMID:25064658

Waszkiewicz, Napoleon; Jelski, Wojciech; Zalewska, Anna; Szulc, Agata; Szmitkowski, Maciej; Zwierz, Krzysztof; Szajda, S?awomir Dariusz

2014-09-01

392

Patterns of salivary gland uptake in I-131 MIBG scintigraphy.  

PubMed

I-131 MIBG scintigraphy is routinely used in the diagnosis of neuroendocrine tumours with high specificity. The radiopharmaceutical is taken up via uptake mechanism and actively transported into storage vesicules. The organs with dense sympathetic innervation such as salivary glands, heart, lachrymal glands, spleen and rarely adrenal medulla are normally visualized with I-131 MIBG. Asymetrical salivary gland uptake is important in a patient with suspected neuroendocrine tumours. Absence of radioactivity may be a result of sympathic denervation or tumor. Bilateral radioactivity absence is observed usually due to drugs or radiopharmaceutical storage conditions. Detailed examination of cervical region is crucial for localisation of neuroendocrine tumours. Therefore, possible false positives should be kept in mind. PMID:16762276

Salanci, B Volkan; Ergün, E Lay

2006-01-01

393

Barley ?-amylase\\/subtilisin inhibitor. I. Isolation and characterization  

Microsoft Academic Search

A protein inhibitor of endogenous ?-amylase 2 has been isolated from germinated barley by glycogen precipitation followed\\u000a by cation-exchange chromatography. Preliminary kinetic analysis showed a mixed type mechanism of inhibition with an apparent\\u000a Ki of 4×10?8M. The inhibitor formed well-defined complexes with barley malt ?-amylase 2 and co-purified with the ?-amylase by cycloheptaamylose\\u000a affinity chromatography of glycogen precipitates. The inhibitor

John Mundy; IB Svendsen; Jørn Hejgaard

1983-01-01

394

Current developments in salivary diagnostics  

PubMed Central

Salivary diagnostics is an emerging field that has progressed through several important developments in the past decade, including the publication of the human salivary proteome and the infusion of federal funds to integrate nanotechnologies and microfluidic engineering concepts into developing compact point-of-care devices for rapid analysis of this secretion. In this article, we discuss some of these developments and their relevance to the prognosis, diagnosis and management of periodontitis, as an oral target, and cardiovascular disease, as a systemic example for the potential of these biodiagnostics. Our findings suggest that several biomarkers are associated with distinct biological stages of these diseases and demonstrate promise as practical biomarkers in identifying and managing periodontal disease, and acute myocardial infarction. The majority of these studies have progressed through biomarker discovery, with the identified molecules requiring more robust clinical studies to enable substantive validation for disease diagnosis. It is predicted that with continued advances in this field the use of a combination of biomarkers in multiplex panels is likely to yield accurate screening tools for these diagnoses in the near future. PMID:20387312

Foley, Joseph D; Bailey, Alison L; Campell, Charles L; Humphries, Roger L; Christodoulides, Nicolaos; Floriano, Pierre N; Simmons, Glennon; Bhagwandin, Bryon; Jacobson, James W; Redding, Spencer W; Ebersole, Jeffrey L; McDevitt, John T

2010-01-01

395

Purification and Characteristics of an Endogenous ?-Amylase Inhibitor from Barley Kernels 1  

PubMed Central

An inhibitor of malted barley (Hordeum vulgare cv Conquest) ?-amylase II was purified 125-fold from a crude extract of barley kernels by (NH4)2SO4 fractionation, ion exchange chromatography on DEAE-Sephacel, and gel filtration on Bio-Gel P 60. The inhibitor was a protein with an approximate molecular weight of 20,000 daltons and an isoelectric point of 7.3. The protein was homogeneous, as assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Amino acid analysis indicated the presence of about 9 half-cystine residues per mole. The neutral isoelectric point of the inhibitor suggested that some of the apparently acidic residues (glutamic and aspartic) existed in the amide form. The first twenty N-terminal amino acids were sequenced. Some homology appeared to exist between the ?-amylase II inhibitor and trypsin inhibitor from barley. Complex formation between ?-amylase II and the inhibitor was detected by the appearance of a new molecular weight species after gel filtration on Bio-Gel P 100. Enzyme and inhibitor had to be preincubated for 5 min, prior to assaying for enzyme activity before maximum inhibition was attained. Inhibition increased at higher pH values. At pH 5.5, an approximately 1100 molar excess of inhibitor over ?-amylase II produced 40% inhibition, whereas, at pH 8.0, a 1:1 molar ratio of inhibitor to enzyme produced the same degree of inhibition. PMID:16663319

Weselake, Randall J.; MacGregor, Alexander W.; Hill, Robert D.; Duckworth, Harry W.

1983-01-01

396

Biochemical characterization of an extracellular polyextremophilic ?-amylase from the halophilic archaeon Halorubrum xinjiangense.  

PubMed

An extracellular haloalkaliphilic thermostable ?-amylase producing archaeon was isolated from the saltwater Lake Urmia and identified as Halorubrum xinjiangense on the basis of morphological, biochemical, and molecular properties. The enzyme was purified to an electrophoretically homogenous state by 80 % cold ethanol precipitation, followed by affinity chromatography. The concentrated pure amylase was eluted as a single peak on fast protein liquid chromatography. The molecular mass of the purified enzyme was about 60 kDa, with a pI value of 4.5. Maximum amylase activity was at 4 M NaCl or 4.5 M KCl, 70 °C, and pH 8.5. The K m and V max of the enzyme were determined as 3.8 mg ml(-1) and 12.4 U mg(-1), respectively. The pure amylase was stable in the presence of SDS, detergents, and organic solvents. In addition, the enzyme (20 U) hydrolyzed 69 % of the wheat starch after a 2-h incubation at 70 °C in an aqueous/hexadecane two-phase system. PMID:23695659

Moshfegh, Mahsa; Shahverdi, Ahmad Reza; Zarrini, Gholamreza; Faramarzi, Mohammad Ali

2013-07-01

397

Carboxymethyl cellulose-gelatin-silica nanohybrid: an efficient carrier matrix for alpha amylase.  

PubMed

Carboxymethyl cellulose (CMC)-gelatin (G) dual templated polymerization of tetramethoxysilane (TMOS) furnished an efficient hybrid carrier support for alpha amylase. The material has been characterized using FTIR, XRD SEM, TGA and BET studies. The amylase was immobilized at the presynthesized hybrid support by adsorption and the immobilized enzyme was used to optimize the conditions for soluble starch hydrolysis. The immobilization did not change the optimum working pH (pH 5) and temperature (40°C) of the enzymatic reaction. The kinetic parameters of the immobilized (Km=9.970mgmL(-1); Vmax=66.23mgmL(-1)min(-1)) and free amylase (KM=4.0509mgmL(-1), Vmax=4.2909mgmL(-1)min(-1)) indicated that the immobilization has enhanced the catalytic function of diastase alpha amylase. The immobilized enzyme showed higher shelf life as compared to the free enzyme in solution and it could be reused for seven consecutive cycles where 85% of the initial activity was exhibited even in the last cycle. The present material is as efficient as our previously reported material CMC-AgNps-Si. PMID:24709014

Singh, Vandana; Ahmad, Shakeel

2014-06-01

398

Effects of split-dose X irradiation on rat salivary gland function  

SciTech Connect

The effect of a single local dose of 15 Gy on salivary gland function in male Albino Wistar rats was compared with the effect of two doses of 7.5 Gy. The intervals chosen were 0-24 h and 1 week. Before and 1-30 days after the last radiation dose, samples of parotid and submandibular saliva were collected simultaneously after stimulation of the glands with pilocarpine. Irradiation with the single dose resulted in an increased lag phase and potassium concentration, and a decreased flow rate and sodium concentration. The rate of secretion of amylase was decreased during Days 1-6, increased at Day 10, and was decreased again at Day 30. With two dose fractions, substantial dose-sparing effects on lag phase, flow rate, and secretion of amylase were observed for both the very early (0-6 days postirradiation) and later (6-30 days postirradiation) effects. These effects were maximal when the interval between the fractions was 6 h. A significant dose-sparing effect on electrolytes was observed for the later effects only, again with a maximum for the 6-h interval. The dose-sparing observed for the very early effects cannot be explained satisfactorily by repair of sublethal damage (SLD), redistribution of cells over the cell cycle, or repopulation of salivary gland tissue between the doses. In contrast to the earlier dose-sparing effects, the split-dose recovery seen for later damage may be attributed, in part, to SLD repair in providing for greater reproductive survival of intercalated ductal cells and enhanced tissue regeneration.

Vissink, A.; s-Gravenmade, E.J.; Ligeon, E.E.; Konings, A.W. (Department of Radiobiology, University of Groningen (Netherlands))

1991-07-01

399

Amlioration des plantes Polymorphisme de l'? amylase chez le pcher.  

E-print Network

Amélioration des plantes Polymorphisme de l'? amylase chez le pêcher. Ã?tude génétique R Monet-de-la-Maye, France (Reçu le 16 octobre 1990; accepté le 13 mars 1991) Résumé — L'a amylase a été étudiée chez'? amylase et celui responsable de la forme des fleurs; en revanche le locus de l'a amylase est indépendant

Paris-Sud XI, Université de

400

Cloning, sequencing, and expression of the gene encoding extracellular alpha-amylase from Pyrococcus furiosus and biochemical characterization of the recombinant enzyme.  

PubMed Central

The gene encoding the hyperthermophilic extracellular alpha-amylase from Pyrococcus furiosus was cloned by activity screening in Escherichia coli. The gene encoded a single 460-residue polypeptide chain. The polypeptide contained a 26-residue signal peptide, indicating that this Pyrococcus alpha-amylase was an extracellular enzyme. Unlike the P. furiosus intracellular alpha-amylase, this extracellular enzyme showed 45 to 56% similarity and 20 to 35% identity to other amylolytic enzymes of the alpha-amylase family and contained the four consensus regions characteristic of that enzyme family. The recombinant protein was a homodimer with a molecular weight of 100,000, as estimated by gel filtration. Both the dimer and monomer retained starch-degrading activity after extensive denaturation and migration on sodium dodecyl sulfate-polyacrylamide gels. The P. furiosus alpha-amylase was a liquefying enzyme with a specific activity of 3,900 U mg-1 at 98 degrees C. It was optimally active at 100 degrees C and pH 5.5 to 6.0 and did not require Ca2+ for activity or thermostability. With a half-life of 13 h at 98 degrees C, the P. furiosus enzyme was significantly more thermostable than the commercially available Bacillus licheniformis alpha-amylase (Taka-therm). PMID:9293008

Dong, G; Vieille, C; Savchenko, A; Zeikus, J G

1997-01-01

401

Receptors for the Neuropeptides, Myoinhibitory Peptide and SIFamide, in Control of the Salivary Glands of the Blacklegged Tick Ixodes scapularis  

PubMed Central

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

Šimo, Ladislav; Ko?i, Juraj; Park, Yoonseong

2013-01-01

402

Inhibitory effects of medicinal mushrooms on ?-amylase and ?-glucosidase - enzymes related to hyperglycemia.  

PubMed

In Asia, medicinal mushrooms have been popularly used as folk medicine and functional foods. In this study, our aim was to examine the inhibitory effects of six medicinal mushrooms on key enzymes (?-amylase and ?-glucosidase) related to hyperglycemia; chemical profiles of bioactive extracts were also examined. The results showed that the n-hexane extract of Coriolus versicolor had the strongest anti-?-amylase activity, while the n-hexane extract of Grifola frondosa showed the most potent anti-?-glucosidase activity. Compared with acarbose, the anti-?-amylase activity of all mushroom extracts was weaker, however a stronger anti-?-glucosidase activity was noted. GC-MS analysis showed that the magnitude of potency of inhibiting ?-glucosidase activity varied with the levels of oleic acid and linoleic acid present in the extracts. These findings were consistent with the IC50 values of these free fatty acids on inhibiting ?-glucosidase activity. Taken together, this study suggests that oleic acid and linoleic acid could have contributed to the potent anti-?-glucosidase activity of selected medicinal mushrooms. PMID:23396484

Su, Chun-Han; Lai, Min-Nan; Ng, Lean-Teik

2013-04-25

403

Inter organ comparison of amylases and starch content in mungbean seedlings  

Microsoft Academic Search

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

Narinder Kaur; Prabhdeep Kaur; Anil K. Gupta

2001-01-01

404

Cloning and expression of a Bacillus coagulans amylase gene in Escherichia coli  

Microsoft Academic Search

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;

Pierre Cornelis; Colette Digneffe; Karine Willemot

1982-01-01

405

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

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

Tsuji, Akihiko; Nishiyama, Nami; Ohshima, Miki; Maniwa, Saori; Kuwamura, Shuji; Shiraishi, Masataka; Yuasa, Keizo

2014-01-01

406

Molecular cloning and characterization of an ?-amylase cDNA highly expressed in major feeding stages of the coffee berry borer, Hypothenemus hampei.  

PubMed

?-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 1879bp, containing a 1458bp open reading frame, which encodes a predicted protein with 485 amino acid residues, with a predicted molecular mass of 51.2kDa. 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

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

407

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

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

Tsuji, Akihiko; Nishiyama, Nami; Ohshima, Miki; Maniwa, Saori; Kuwamura, Shuji; Shiraishi, Masataka; Yuasa, Keizo

2014-01-01

408

Enzyme Assay Guided Isolation of an ?-Amylase Inhibitor Flavonoid from Vaccinium arctostaphylos Leaves  

PubMed Central

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

Nickavar, Bahman; Amin, Gholamreza

2011-01-01

409