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Performance evaluation of salivary amylase activity monitor.
Yamaguchi, Masaki; Kanemori, Takahiro; Kanemaru, Masashi; Takai, Noriyasu; Mizuno, Yasufumi; Yoshida, Hiroshi
In order to quantify psychological stress and to distinguish eustress and distress, we have been investigating the establishment of a method that can quantify salivary amylase activity (SMA). Salivary glands not only act as amplifiers of a low level of norepinephrine, but also respond more quickly and sensitively to psychological stress than cortisol levels. Moreover, the time-course changes of the salivary amylase activity have a possibility to distinguish eustress and distress. Thus, salivary amylase activity can be utilized as an excellent index for psychological stress. However, in dry chemistry system, a method for quantification of the enzymatic activity still needs to be established that can provide with sufficient substrate in a testing tape as well as can control enzymatic reaction time. Moreover, it is necessary to develop a method that has the advantages of using saliva, such as ease of collection, rapidity of response, and able to use at any time. In order to establish an easy method to monitor the salivary amylase activity, a salivary transcription device was fabricated to control the enzymatic reaction time. A fabricated salivary amylase activity monitor consisted of three devices, the salivary transcription device, a testing-strip and an optical analyzer. By adding maltose as a competitive inhibitor to a substrate Ga1-G2-CNP, a broad-range activity testing-strip was fabricated that could measure the salivary amylase activity with a range of 0-200 kU/l within 150 s. The calibration curve of the monitor for the salivary amylase activity showed R2=0.941, indicating that it was possible to use this monitor for the analysis of the salivary amylase activity without the need to determine the salivary volume quantitatively. In order to evaluate the assay variability of the monitor, salivary amylase activity was measured using Kraepelin psychodiagnostic test as a psychological stressor. A significant difference of salivary amylase activity was recognized between the pre-stress and mid-stress periods. This study demonstrated that broad-range salivary amylase activity monitor was developed that could be used with only 5 microl of saliva. PMID:15494230
Performance evaluation of salivary amylase activity monitor
Masaki Yamaguchi; Takahiro Kanemori; Masashi Kanemaru; Noriyasu Takai; Yasufumi Mizuno; Hiroshi Yoshida
In order to quantify psychological stress and to distinguish eustress and distress, we have been investigating the establishment of a method that can quantify salivary amylase activity (SMA). Salivary glands not only act as amplifiers of a low level of norepinephrine, but also respond more quickly and sensitively to psychological stress than cortisol levels. Moreover, the time-course changes of the
April 16, 2010 Caffeine and Stress Alter Salivary -Amylase Activity in Young Men
Ritter, Frank
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
Caffeine and stress alter salivary a-amylase activity in young men Laura C. Klein1*, Jeanette M. Bennett1
Ritter, Frank
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
Possible involvement of ?? receptors in various emetogen-induced increases in salivary amylase activity in rats.
Fukui, Hideo; Suyama, Yoshimi; Iwachido, Takako; Miwa, Eri
We investigated the inhibitory effects of ??- or ??-adrenoceptor (AR) antagonists on salivary amylase secretion produced by various emetic agents, such as cisplatin, apomorphine, and lithium chloride (LiCl), or the non-emetic agent ?(½)-AR agonist isoprenaline in rats. We also determined the inhibitory effect of metoclopramide, a dopamine D?-receptor antagonist, on increases in the salivary amylase activity induced by apomorphine or granisetron, a 5-HT(3)-receptor antagonist, on LiCl-induced increased salivary amylase activity. Isoprenaline (0.01 mg/kg, s.c.) produced an increase in salivary amylase and the increase was inhibited by the ?(½)-AR antagonist propranolol (5 mg/kg, s.c.) and ??-AR antagonist atenolol (2 mg/kg, s.c.) but not by the ??-AR antagonist butoxamine (8 mg/kg, s.c.). The increased amylase activity induced by cisplatin (15 mg/kg, i.v.), apomorphine (3 mg/kg, s.c.), or LiCl (120 mg/kg, i.p.) was inhibited significantly by atenolol (2 mg/kg, s.c.) but not by butoxamine (8 mg/kg, s.c.). In addition, increases in amylase activities induced by apomorphine and LiCl were inhibited significantly by metoclopramide (10 mg/kg, i.v.) and granisetron (3 mg/kg, i.v.), respectively. These results suggest that salivary amylase secretion induced by various emetogens is involved in ??-adrenoceptor activity and that salivary amylase activity is useful to detect emetogens with no direct ??-AR activation in rats, a species that does not exhibit vomiting. PMID:21173550
Enhancing Maritime Education and Training: Measuring a Ship Navigator's Stress Based on Salivary Amylase Activity
ERIC Educational Resources Information Center
Murai, Koji; Wakida, Shin-Ichi; Miyado, Takashi; Fukushi, Keiichi; Hayashi, Yuji; Stone, Laurie C.
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…
The relationship between the level of salivary alpha amylase activity and pain severity in patients with symptomatic irreversible pulpitis
Shahriari, Shahriar; Goodarzi, Mohammad Taghi; Moghimbeigi, Abbas; Jazaeri, Mina; Babaei, Parisa
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
Purification of a novel ?-amylase inhibitor from local Himalayan bean (Phaseolus vulgaris) seeds with activity towards bruchid pests and human salivary amylase.
Gupta, Mridu; Sharma, Pratima; Nath, Amarjit K
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
Oral Fusobacterium nucleatum subsp. polymorphum binds to human salivary ?-amylase.
Zulfiqar, M; Yamaguchi, T; Sato, S; Oho, T
Fusobacterium nucleatum acts as an intermediate between early and late colonizers in the oral cavity. In this study, we showed that F. nucleatum subsp. polymorphum can bind to a salivary component with a molecular weight of approximately 110 kDa and identified the protein and another major factor of 55 kDa, as salivary ?-amylase by time-of-flight mass spectrometry and immuno-reactions. Salivary ?-amylase is present in both monomeric and dimeric forms and we found that formation of the dimer depends on copper ions. The F. nucleatum adhered to both monomeric and dimeric salivary ?-amylases, but the numbers of bacteria bound to the dimeric form were more than those bound to the monomeric form. The degree of adherence of F. nucleatum to four ?-amylases from different sources was almost the same, however its binding to ?-amylase was considerably decreased. Among four ?-amylase inhibitors tested, acarbose and type 1 and 3 inhibitors derived from wheat flour showed significant activity against the adhesion of F.nucleatum to monomeric and dimeric amylases, however voglibose had little effect. Moreover F. nucleatum cells inhibited the enzymatic activity of salivary ?-amylase in a dose-dependent manner. These results suggest that F. nucleatum plays more important and positive role as an early colonizer for maturation of oral microbial colonization. PMID:23906425
Effects of methamphetamine on the noradrenergic activity biomarker salivary alpha-amylase*
Haile, Colin N.; De La Garza, Richard; Mahoney, James J.; Newton, Thomas F.
Background Methamphetamine (METH) potently activates the sympathetic nervous system (SNS) by increasing central and peripheral norepinephrine (NE). Salivary ?-amylase (sAA) is a biomarker of SNS activation that correlates with plasma NE levels. The purpose of this study was to determine the impact of METH on sAA activity and whether changes in sAA activity were correlated with subjective effects ratings. Methods Non-treatment seeking METH-dependent volunteers (N=8) participated in this within-subjects laboratory-based study. Volunteers received randomly administered intravenous METH (0mg, 30mg) and sAA activity, cardiovascular measures and subjective ratings were assessed at baseline (?15 min) and five post-METH time points (10, 20, 30, 45, and 60 min). Results METH (30mg) increased sAA activity over time. sAA activity significantly correlated with diastolic blood pressure following 0mg METH and systolic blood pressure following 30mg METH. Subjective ratings (ANY EFFECT, HIGH, GOOD, STIMULATED, LIKE, WLLING TO PAY) highly correlated with sAA over five post-METH time points (N=40; r's=0.543–0.684, p's <0.001). Age, body mass index and METH amount received on a mg/kg basis were significantly associated with sAA activity. Multiple linear regression analysis indicated sAA activity remained a significant predictor of subjective ratings following METH after controlling for these factors. Conclusions The NE peripheral biomarker sAA activity is associated with METH's subjective effects. PMID:23968815
Psychosocial Stress-Induced Activation of Salivary Alpha-Amylase: An Indicator of Sympathetic Activity?
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-
Salivary ?-amylase, serum albumin, and myoglobin protect against DNA-damaging activities of ingested dietary agents in vitro.
Hossain, M Zulfiquer; Patel, Kalpesh; Kern, Scott E
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
Coordination of murine parotid secretory protein and salivary amylase expression.
Poulsen, K; Jakobsen, B K; Mikkelsen, B M; Harmark, K; Nielsen, J T; Hjorth, J P
PSP, parotid secretory protein, and salivary amylase are the major secretory proteins of mouse parotid gland where they appear in a constant ratio. Here we describe the isolation of the PSP gene and show through expression analysis on this and the salivary amylase gene that the two genes are transcribed in a coordinate fashion in adult animals, whereas the activation profiles are different during postnatal development. An explanation is put forward that involves activation of the genes at different stages of the acinar cell differentiation, leading in adults to the maximal and thus proportionate expression. Images Fig. 4. Fig. 5. PMID:2428613
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
Ritter, Frank
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
Characterization of salivary alpha-amylase binding to Streptococcus sanguis
Scannapieco, F.A.; Bergey, E.J.; Reddy, M.S.; Levine, M.J. (State Univ. of New York, Buffalo (USA))
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.
Characterization of salivary alpha-amylase binding to Streptococcus sanguis.
Scannapieco, F A; Bergey, E J; Reddy, M S; Levine, M J
The purpose of this study was to identify the major salivary components which interact with oral bacteria and to determine the mechanism(s) responsible for their binding to the bacterial surface. Strains of Streptococcus sanguis, Streptococcus mitis, Streptococcus mutans, and Actinomyces viscosus were incubated for 2 h in freshly collected human submandibular-sublingual saliva (HSMSL) or parotid saliva (HPS), and bound salivary components were eluted with 2% sodium dodecyl sulfate. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western transfer, alpha-amylase (EC 126.96.36.199) was the prominent salivary component eluted from S. sanguis. Studies with 125I-labeled HSMSL or 125I-labeled HPS also demonstrated a component with an electrophoretic mobility identical to that of alpha-amylase which bound to S. sanguis. Purified alpha-amylase from human parotid saliva was radiolabeled and found to bind to strains of S. sanguis genotypes 1 and 3 and S. mitis genotype 2, but not to strains of other species of oral bacteria. Binding of [125I]alpha-amylase to streptococci was saturable, calcium independent, and inhibitable by excess unlabeled alpha-amylases from a variety of sources, but not by secretory immunoglobulin A and the proline-rich glycoprotein from HPS. Reduced and alkylated alpha-amylase lost enzymatic and bacterial binding activities. Binding was inhibited by incubation with maltotriose, maltooligosaccharides, limit dextrins, and starch. Images PMID:2788139
Characterization of salivary alpha-amylase binding to Streptococcus sanguis.
Scannapieco, F A; Bergey, E J; Reddy, M S; Levine, M J
The purpose of this study was to identify the major salivary components which interact with oral bacteria and to determine the mechanism(s) responsible for their binding to the bacterial surface. Strains of Streptococcus sanguis, Streptococcus mitis, Streptococcus mutans, and Actinomyces viscosus were incubated for 2 h in freshly collected human submandibular-sublingual saliva (HSMSL) or parotid saliva (HPS), and bound salivary components were eluted with 2% sodium dodecyl sulfate. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western transfer, alpha-amylase (EC 22.214.171.124) was the prominent salivary component eluted from S. sanguis. Studies with 125I-labeled HSMSL or 125I-labeled HPS also demonstrated a component with an electrophoretic mobility identical to that of alpha-amylase which bound to S. sanguis. Purified alpha-amylase from human parotid saliva was radiolabeled and found to bind to strains of S. sanguis genotypes 1 and 3 and S. mitis genotype 2, but not to strains of other species of oral bacteria. Binding of [125I]alpha-amylase to streptococci was saturable, calcium independent, and inhibitable by excess unlabeled alpha-amylases from a variety of sources, but not by secretory immunoglobulin A and the proline-rich glycoprotein from HPS. Reduced and alkylated alpha-amylase lost enzymatic and bacterial binding activities. Binding was inhibited by incubation with maltotriose, maltooligosaccharides, limit dextrins, and starch. PMID:2788139
Various emetogens increase the secretion of salivary amylase in rats: a potential model in emesis research.
Fukui, Hideo; Miwa, Eri; Iwachido, Takako; Kitaura, Harumi; Furukawa, Hatsue
We investigated the effects of various emetic agents: cisplatin, apomorphine, lithium chloride (LiCl), rolipram, sibutramine, and the beta(3)-adrenoceptor (AR) agonist CL316243 on salivary amylase secretion in rats. We also determined the inhibitory effect of granisetron, a 5-HT(3)-receptor antagonist, on cisplatin-induced increased salivary amylase activity and the inhibitory effect of bilateral abdominal vagotomy on increases in salivary amylase activity induced by cisplatin and LiCl. Granisetron was administered 15 min before and 1 h after cisplatin administration. Cisplatin (10 - 15 mg/kg, i.v.) increased salivary amylase activity dose-dependently and induced an acute increase from 1.5 h post-treatment with 15 mg/kg. Apomorphine (1 - 3 mg/kg, s.c.), LiCl (120 mg/kg, i.p.), rolipram (3 - 10 mg/kg, p.o.), and sibutramine (10 mg/kg, p.o.) induced significant increases in salivary amylase secretion. On the other hand, CL316243 did not stimulate salivary amylase secretion. The increased amylase activity induced by cisplatin (15 mg/kg, i.v.) was inhibited significantly by granisetron (1 or 3 mg/kg x 2, i.v.) or tended to be inhibited by bilateral abdominal vagotomy, whereas an increase in amylase activity produced by LiCl was not inhibited by abdominal visceral nerve section. These results suggest that salivary amylase activity is useful as a marker for emesis in rats, a species that does not exhibit vomiting. PMID:20484866
Effect of an herb root extract, herbal dentifrice and synthetic dentifrice on human salivary amylase
Sapra, Gaurav; Vyas, Yogesh Kumar; Agarwal, Rahul; Aggarwal, Ashish; Chandrashekar, K T; Sharma, Kanika
Background: Salivary amylase is an enzyme, which plays a vital role in formation of dental plaque. It has the ability to bind on the bacterial surfaces and to hydrolyze starch, giving rise to products that are transformed into acids leading to dental caries. Suppression of salivary amylase activity can lead to decrease in risk of dental caries and plaque associated periodontal diseases. The aim of this study was to evaluate the effect of an herb, Spilanthes calva (in form of a test dentifrice) on human salivary amylase activity and to compare it with other dentifrices. Materials and Methods: A total of 80 subjects of age 18-35 years were randomly selected and divided equally into 4 groups. Group 1 subjects were assigned to use Test Dentifrice (with S. calva root extract), while Group 2, Group 3, and Group 4 subjects were assigned to use Herbal Dentifrice (Arodent™), Synthetic Dentifrice (Colgate®), and Control Dentifrice respectively. Salivary amylase activity was determined by Bernfeld method in each group, before and after using the given dentifrices. Results: Maximum inhibition of salivary amylase activity was found in the group using test dentifrice as compared to others. Conclusion: The present study indicates that, the root extract of S. calva possess significant inhibitory activity for salivary amylase. Use of S. calva root extract will provide a wider protection against different pathogenic oral microflora. Use of this extract singly or in combination is strongly recommended in the dentifrice formulations. PMID:24130585
Structural relationship between the enzymatic and streptococcal binding sites of human salivary alpha-amylase.
Scannapieco, F A; Bhandary, K; Ramasubbu, N; Levine, M J
Previous studies have demonstrated that human salivary alpha-amylase specifically binds to the oral bacterium Streptococcus gordonii. This interaction is inhibited by substrates such as starch and maltotriose suggesting that bacterial binding may involve the enzymatic site of amylase. Experiments were performed to determine if amylase bound to the bacterial surface possessed enzymatic activity. It was found that over one-half of the bound amylase was enzymatically active. In addition, bacterial-bound amylase hydrolyzed starch to glucose which was then metabolized to lactic acid by the bacteria. In further studies, the role of amylase's histidine residues in streptococcal binding and enzymatic function was assessed after their selective modification with diethyl pyrocarbonate. DEP-modified amylase showed a marked reduction in both enzymatic and streptococcal binding activities. These effects were diminished when DEP modification occurred in the presence of maltotriose. DEP-modified amylase had a significantly altered secondary structure when compared with native enzyme or amylase modified in the presence of maltotriose. Collectively, these results suggest that human salivary alpha-amylase may possess multiple sites for bacterial binding and enzymatic activity which share structural similarities. PMID:2125215
Anaerobic Threshold and Salivary ?-amylase during Incremental Exercise.
Akizuki, Kazunori; Yazaki, Syouichirou; Echizenya, Yuki; Ohashi, Yukari
[Purpose] The purpose of this study was to clarify the validity of salivary ?-amylase as a method of quickly estimating anaerobic threshold and to establish the relationship between salivary ?-amylase and double-product breakpoint in order to create a way to adjust exercise intensity to a safe and effective range. [Subjects and Methods] Eleven healthy young adults performed an incremental exercise test using a cycle ergometer. During the incremental exercise test, oxygen consumption, carbon dioxide production, and ventilatory equivalent were measured using a breath-by-breath gas analyzer. Systolic blood pressure and heart rate were measured to calculate the double product, from which double-product breakpoint was determined. Salivary ?-amylase was measured to calculate the salivary threshold. [Results] One-way ANOVA revealed no significant differences among workloads at the anaerobic threshold, double-product breakpoint, and salivary threshold. Significant correlations were found between anaerobic threshold and salivary threshold and between anaerobic threshold and double-product breakpoint. [Conclusion] As a method for estimating anaerobic threshold, salivary threshold was as good as or better than determination of double-product breakpoint because the correlation between anaerobic threshold and salivary threshold was higher than the correlation between anaerobic threshold and double-product breakpoint. Therefore, salivary threshold is a useful index of anaerobic threshold during an incremental workload. PMID:25140097
Words number: 25491 Assessment of salivary alpha-amylase -a stress biomarker -in2
Boyer, Edmond
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
Salivary amylase promotes adhesion of oral streptococci to hydroxyapatite.
Scannapieco, F A; Torres, G I; Levine, M J
Recent studies have demonstrated that several species of oral streptococci, such as Streptococcus gordonii, bind soluble salivary alpha-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 ability to promote adhesion of S. gordonii to hydroxyapatite. Fractions containing alpha-amylase and proline-rich proteins promoted the adhesion of [3H]-labeled S. gordonii to hydroxyapatite. Similar findings were obtained with purified amylase and acidic proline-rich protein 1 (PRP1). Incubation of S. gordonii G9B in the presence of starch and maltotriose increased the binding of this strain to amylase-coated hydroxyapatite, while the adhesion of S. sanguis 10556 to amylase-coated hydroxyapatite was not affected by these saccharides. These results suggest that amylase may serve as a hydroxyapatite pellicle receptor for amylase-binding streptococci. Furthermore, starch and starch metabolites may enhance the adhesion of amylase-binding streptococci to amylase in dental pellicles to augment the formation of dental plaque. PMID:7560386
Salivary Amylase Promotes Adhesion of Oral Streptococci to Hydroxyapatite
F. A. Scannapieco; G. I. Torres; M. J. Levine
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
Susceptibility to corrosion of laser welding composite arch wire in artificial saliva of salivary amylase and pancreatic amylase.
Zhang, Chao; Liu, Jiming; Yu, Wenwen; Sun, Daqian; Sun, Xinhua
In this study, laser-welded composite arch wire (CAW) with a copper interlayer was exposed to artificial saliva containing salivary amylase or pancreatic amylase, and the resultant corrosion behavior was studied. The purpose was to determine the mechanisms by which salivary amylase and pancreatic amylase contribute to corrosion. The effects of amylase on the electrochemical resistance of CAW were tested by potentiodynamic polarization measurements. The dissolved corrosion products were determined by ICP-OES, and the surfaces were analyzed by SEM, AFM and EDS. The results showed that both exposure to salivary amylase and pancreatic amylase significantly improved the corrosion resistance of CAW. Even isozyme could have different influences on the alloy surface. When performing in vitro research of materials to be used in oral cavity, the effect of ?-amylase should be taken into account since a simple saline solution does not entirely simulate the physiological situation. PMID:26117761
Salivary ?-amylase and cortisol after exercise in menopause: influence of long-term HRT.
Patacchioli, F R; Ghiciuc, C M; Bernardi, M; Dima-Cozma, L C; Fattorini, L; Squeo, M R; Galoppi, P; Brunelli, R; Ferrante, F; Pasquali, V; Perrone, G
Objectives This observational prospective study analyzed the effect of an incremental cardiopulmonary exercise test (CPET) on the secretion of salivary biomarkers of the adrenergic nervous system and hypothalamus-pituitary-adrenal (HPA) axis activity by measuring salivary ?-amylase and cortisol diurnal trajectories in the setting of long-term hormone replacement therapy (HRT). Methods Fifteen healthy sedentary postmenopausal women who were current HRT users and 15 women who had never used HRT were consecutively recruited. ?-Amylase and cortisol were measured in salivary samples collected on the CPET day and on a rest day. Cardiovascular and respiratory fitness parameters were recorded during the CPET challenge. Results The participants had very homogeneous somatic characteristics, and they were all in generally good health. The postmenopausal never-HRT users presented an abnormal diurnal pattern of ?-amylase at baseline and a flattened response to CPET. In contrast, women on HRT had a physiological ?-amylase diurnal pattern and increased salivary ?-amylase production during the CPET-induced challenge. The CPET challenge physiologically activated the HPA axis activity, as shown by the increase in the concentration of salivary cortisol during the effort test. HPA axis activity was not affected by long-term HRT. Postmenopausal women using HRT exhibited a cardiorespiratory functional capacity that was significantly (p < 0.05) higher than that of non-users. Conclusions Our findings show that healthy postmenopausal women present an asymmetry between adrenergic nervous system and HPA axis activities under both basal and stress conditions. HRT was able to modify the abnormal adrenergic nervous system activity, most likely by reducing the sympathetic hyperactivity that characterizes menopause. PMID:25602168
Human salivary alpha-amylase reactivity in a psychosocial stress paradigm
Urs M. Nater; Nicolas Rohleder; Jens Gaab; Simona Berger; Andreas Jud; Clemens Kirschbaum; Ulrike Ehlert
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
Salivary alpha-amylase: role in dental plaque and caries formation.
Scannapieco, F A; Torres, G; Levine, M J
Salivary alpha-amylase, one of the most plentiful components in human saliva, has at least three distinct biological functions. The enzymatic activity of alpha-amylase undoubtedly plays a role in carbohydrate digestion. Amylase in solution binds with high affinity to a selected group of oral streptococci, a function that may contribute to bacterial clearance and nutrition. The fact that alpha-amylase is also found in acquired enamel pellicle suggests a role in the adhesion of alpha-amylase-binding bacteria. All of these biological activities seem to depend on an intact enzyme conformation. Binding of alpha-amylase to bacteria and teeth may have important implications for dental plaque and caries formation. alpha-Amylase bound to bacteria in plaque may facilitate dietary starch hydrolysis to provide additional glucose for metabolism by plaque microorganisms in close proximity to the tooth surface. The resulting lactic acid produced may be added to the pool of acid in plaque to contribute to tooth demineralization. PMID:8373987
Salivary ?-Amylase Reactivity to Infant Crying in Maltreating Mothers.
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
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
Interaction of Salivary alpha-Amylase and Amylase-Binding-Protein A (AbpA) of Streptococcus gordonii with Glucosyltransferase of S. gordonii and Streptococcus mutans
Background Glucosyltransferases (Gtfs), enzymes that produce extracellular glucans from dietary sucrose, contribute to dental plaque formation by Streptococcus gordonii and Streptococcus mutans. The alpha-amylase-binding protein A (AbpA) of S. gordonii, an early colonizing bacterium in dental plaque, interacts with salivary amylase and may influence dental plaque formation by this organism. We examined the interaction of amylase and recombinant AbpA (rAbpA), together with Gtfs of S. gordonii and S. mutans. Results The addition of salivary alpha-amylase to culture supernatants of S. gordonii precipitated a protein complex containing amylase, AbpA, amylase-binding protein B (AbpB), and the glucosyltransferase produced by S. gordonii (Gtf-G). rAbpA was expressed from an inducible plasmid, purified from Escherichia coli and characterized. Purified rAbpA, along with purified amylase, interacted with and precipitated Gtfs from culture supernatants of both S. gordonii and S. mutans. The presence of amylase and/or rAbpA increased both the sucrase and transferase component activities of S. mutans Gtf-B. Enzyme-linked immunosorbent assay (ELISA) using anti-Gtf-B antibody verified the interaction of rAbpA and amylase with Gtf-B. A S. gordonii abpA-deficient mutant showed greater biofilm growth under static conditions than wild-type in the presence of sucrose. Interestingly, biofilm formation by every strain was inhibited in the presence of saliva. Conclusion The results suggest that an extracellular protein network of AbpA-amylase-Gtf may influence the ecology of oral biofilms, likely during initial phases of colonization. PMID:17593303
Elevated salivary ?-amylase and cortisol levels in unremitted and remitted depressed patients.
Ishitobi, Yoshinobu; Akiyoshi, Jotaro; Tanaka, Yoshihiro; Ando, Tomoko; Okamoto, Shizuko; Kanehisa, Masayuki; Kohno, Kentaro; Ninomiya, Taiga; Maruyama, Yoshihiro; Tsuru, Jusen; Kawano, Aimi; Hanada, Hiroaki; Isogawa, Koichi; Kodama, Kensuke
Abstract Objective. Major depressive disorder (MDD) is often associated with dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis via chronic stress. Psychosocial stress-induced activation of salivary ?-amylase (sAA) represents sympathoadrenal medullary system (SAM) activity, and sAA has become an emerging biomarker for sympathetic nervous system activity. In contrast to salivary cortisol, sAA has been less extensively studied in depressed patients. The present study sought to address this problem by measuring sAA and salivary cortisol levels in patients with major depressive disorder. Methods. The authors recorded Spielberger State-Trait Anxiety Inventory (STAI) scores along with, levels of sAA and salivary cortisol in 28 patients with unremitted major depressive disorder, 43 remitted patients and 103 healthy volunteers. Results. STAI (State or Trait) measurements in unremitted patients with MDD were significantly increased compared with healthy controls and remitted patients. SAA and cortisol levels in unremitted patients were also significantly elevated compared to controls and remitted patients. Finally, sAA levels were significantly correlated with HRSD in unremitted patients with MDD. Conclusion. These preliminary results suggest that sAA may be a state-dependent marker of major depressive disorder in addition to salivary cortisol. PMID:24917438
Sex differences in salivary cortisol, alpha-amylase, and psychological functioning following Hurricane Katrina.
Vigil, Jacob M; Geary, David C; Granger, Douglas A; Flinn, Mark V
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 alpha-amylase, depression, anxiety, distress, aggression, and self-esteem for this group were contrasted with a demographically matched no-trauma control group (n = 53). Results revealed that hurricane exposure and SNS activity moderated the relations between lower cortisol and higher internalizing behaviors. Sex-related differences were observed in behavioral adjustment and stress regulation. Implications of sex differences in biobehavioral adjustment to loss, displacement, and relocation are discussed in relation to evolutionary and developmental theory. PMID:20636692
Salivary alpha-amylase and cortisol responsiveness following electrically stimulated physical stress in bipolar disorder patients
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
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
Response of fatty acid synthesis genes to the binding of human salivary amylase by Streptococcus gordonii.
Nikitkova, Anna E; Haase, Elaine M; Vickerman, M Margaret; Gill, Steven R; Scannapieco, Frank A
Streptococcus gordonii, an important primary colonizer of dental plaque biofilm, specifically binds to salivary amylase via the surface-associated amylase-binding protein A (AbpA). We hypothesized that a function of amylase binding to S. gordonii may be to modulate the expression of chromosomal genes, which could influence bacterial survival and persistence in the oral cavity. Gene expression profiling by microarray analysis was performed to detect genes in S. gordonii strain CH1 that were differentially expressed in response to the binding of purified human salivary amylase versus exposure to purified heat-denatured amylase. Selected genes found to be differentially expressed were validated by quantitative reverse transcription-PCR (qRT-PCR). Five genes from the fatty acid synthesis (FAS) cluster were highly (10- to 35-fold) upregulated in S. gordonii CH1 cells treated with native amylase relative to those treated with denatured amylase. An abpA-deficient strain of S. gordonii exposed to amylase failed to show a response in FAS gene expression similar to that observed in the parental strain. Predicted phenotypic effects of amylase binding to S. gordonii strain CH1 (associated with increased expression of FAS genes, leading to changes in fatty acid synthesis) were noted; these included increased bacterial growth, survival at low pH, and resistance to triclosan. These changes were not observed in the amylase-exposed abpA-deficient strain, suggesting a role for AbpA in the amylase-induced phenotype. These results provide evidence that the binding of salivary amylase elicits a differential gene response in S. gordonii, resulting in a phenotypic adjustment that is potentially advantageous for bacterial survival in the oral environment. PMID:22247133
Response of Fatty Acid Synthesis Genes to the Binding of Human Salivary Amylase by Streptococcus gordonii
Nikitkova, Anna E.; Haase, Elaine M.; Vickerman, M. Margaret; Gill, Steven R.
Streptococcus gordonii, an important primary colonizer of dental plaque biofilm, specifically binds to salivary amylase via the surface-associated amylase-binding protein A (AbpA). We hypothesized that a function of amylase binding to S. gordonii may be to modulate the expression of chromosomal genes, which could influence bacterial survival and persistence in the oral cavity. Gene expression profiling by microarray analysis was performed to detect genes in S. gordonii strain CH1 that were differentially expressed in response to the binding of purified human salivary amylase versus exposure to purified heat-denatured amylase. Selected genes found to be differentially expressed were validated by quantitative reverse transcription-PCR (qRT-PCR). Five genes from the fatty acid synthesis (FAS) cluster were highly (10- to 35-fold) upregulated in S. gordonii CH1 cells treated with native amylase relative to those treated with denatured amylase. An abpA-deficient strain of S. gordonii exposed to amylase failed to show a response in FAS gene expression similar to that observed in the parental strain. Predicted phenotypic effects of amylase binding to S. gordonii strain CH1 (associated with increased expression of FAS genes, leading to changes in fatty acid synthesis) were noted; these included increased bacterial growth, survival at low pH, and resistance to triclosan. These changes were not observed in the amylase-exposed abpA-deficient strain, suggesting a role for AbpA in the amylase-induced phenotype. These results provide evidence that the binding of salivary amylase elicits a differential gene response in S. gordonii, resulting in a phenotypic adjustment that is potentially advantageous for bacterial survival in the oral environment. PMID:22247133
Salivary alpha-amylase and cortisol responsiveness following electrical stimulation stress in major depressive disorder patients.
Tanaka, Yoshihiro; Ishitobi, Yoshinobu; Maruyama, Yoshihiro; Kawano, Aimi; Ando, Tomoko; Okamoto, Shizuko; Kanehisa, Masayuki; Higuma, Haruka; Ninomiya, Taiga; Tsuru, Jusen; Hanada, Hiroaki; Kodama, Kensuke; Isogawa, Koichi; Akiyoshi, Jotaro
Major depressive disorder (MDD) is often associated with dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis by chronic stress. In comparison, psychosocial stress-induced activation of salivary ?-amylase (sAA) functions as a marker of sympathoadrenal medullary system (SAM) activity. However, in contrast to salivary cortisol, sAA has been less extensively studied in MDD patients. The present study measured sAA and salivary cortisol levels in patients with MDD. The authors determined Profile of Mood State (POMS) and State-Trait anxiety Inventory (STAI) scores, Heart Rate Variability (HRV), and sAA and salivary cortisol levels in 88 patients with MDD and 41 healthy volunteers following the application of electrical stimulation stress. Patients with major depressive disorder were 8 points or more on Hamilton Depression Scale (HAM-D) scores. Tension-Anxiety, Depression-Dejection, Anger-Hostility, Fatigue, and Confusion scores in patients with major depressive disorder were significantly increased compared to healthy controls. In contrast, Vigor scores in patients with MDD were significantly decreased compared with healthy controls. There was no difference in heart rate variability measures between MDD patients and healthy controls. The threshold of electrical stimulation applied in MDD patients was lower than that in healthy controls. SAA levels in female MDD patients were significantly elevated relative to controls both before and after electrical stimulation. Finally, there were no differences in salivary cortisol levels between major depressive patients and controls. In the present study only three time points were explored. Furthermore, the increased secretion of sAA before and after stimulation could allude to an increased responsiveness of novel and uncontrollable situations in patients with MDD. These preliminary results suggest that sAA might be a useful biological marker of MDD. PMID:22063648
The psychosocial stress-induced increase in salivary alpha-amylase is independent of saliva flow rate
Nicolas Rohleder; Jutta M. Wolf; Enrique F. Maldonado; Clemens Kirschbaum
The stress response of salivary alpha-amylase (sAA) has been suggested as an index for sympathetic nervous system activation. However, concurrent inhibition of the parasympathetic nervous system is discussed as a confounder due to suppression of saliva flow rate. Here we set out to test the influence of stress-induced changes in flow rate on sAA secretion. Twenty-six subjects underwent the Trier
Structure of amylase-binding protein A of Streptococcus gordonii: A potential receptor for human salivary ?-amylase enzyme.
Sethi, Ashish; Mohanty, Biswaranjan; Ramasubbu, Narayanan; Gooley, Paul R
Amylase-binding protein A (AbpA) of a number of oral streptococci is essential for the colonization of the dental pellicle. We have determined the solution structure of residues 24-195 of AbpA of Streptococcus gordonii and show a well-defined core of five helices in the region of 45-115 and 135-145. (13) C?/? chemical shift and heteronuclear (15) N-{(1) H} NOE data are consistent with this fold and that the remainder of the protein is unstructured. The structure will inform future molecular experiments in defining the mechanism of human salivary ?-amylase binding and biofilm formation by streptococci. PMID:25739638
Discovering an Accessible Enzyme: Salivary [alpha]-Amylase--"Prima Digestio Fit in Ore"--A Didactic Approach for High School Students
ERIC Educational Resources Information Center
Marini, Isabella
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…
Evaluation of autonomic nervous system by salivary alpha-amylase level and heart rate variability in patients with schizophrenia.
Ieda, Masa; Miyaoka, Tsuyoshi; Wake, Rei; Liaury, Kristian; Tsuchie, Keiko; Fukushima, Michiyo; Araki, Tomoko; Ezoe, Satoko; Inagaki, Takuji; Horiguchi, Jun
Several researches indicate that autonomic nervous system (ANS) dysfunction in patients with schizophrenia. Recently, salivary alpha-amylase (sAA) has been employed as a useful marker for ANS function. We investigated the extent of ANS dysfunction by measuring sAA and heart rate variability (HRV) of 25 patients with schizophrenia compared with controls. Schizophrenia group demonstrated a significant increase in sAA and markedly lower parasympathetic nervous system (PNS) activity in the HRV. However, there were no significant differences between two groups in sympathetic nervous system (SNS) activity. We concluded that PNS might be suppressed and the SNS shows relatively high activity in schizophrenia. PMID:23645102
Salivary alpha-amylase and cortisol responsiveness following electrical stimulation stress in obsessive-compulsive disorder patients.
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
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
Differences in Salivary Alpha-Amylase and Cortisol Responsiveness following Exposure to Electrical Stimulation versus the Trier Social Stress Tests
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
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
Taking the starch out of oral biofilm formation: molecular basis and functional significance of salivary ?-amylase binding to oral streptococci.
Nikitkova, Anna E; Haase, Elaine M; Scannapieco, Frank A
?-Amylase-binding streptococci (ABS) are a heterogeneous group of commensal oral bacterial species that comprise a significant proportion of dental plaque microfloras. Salivary ?-amylase, one of the most abundant proteins in human saliva, binds to the surface of these bacteria via specific surface-exposed ?-amylase-binding proteins. The functional significance of ?-amylase-binding proteins in oral colonization by streptococci is important for understanding how salivary components influence oral biofilm formation by these important dental plaque species. This review summarizes the results of an extensive series of studies that have sought to define the molecular basis for ?-amylase binding to the surface of the bacterium as well as the biological significance of this phenomenon in dental plaque biofilm formation. PMID:23144140
Taking the Starch out of Oral Biofilm Formation: Molecular Basis and Functional Significance of Salivary ?-Amylase Binding to Oral Streptococci
Nikitkova, Anna E.; Haase, Elaine M.
?-Amylase-binding streptococci (ABS) are a heterogeneous group of commensal oral bacterial species that comprise a significant proportion of dental plaque microfloras. Salivary ?-amylase, one of the most abundant proteins in human saliva, binds to the surface of these bacteria via specific surface-exposed ?-amylase-binding proteins. The functional significance of ?-amylase-binding proteins in oral colonization by streptococci is important for understanding how salivary components influence oral biofilm formation by these important dental plaque species. This review summarizes the results of an extensive series of studies that have sought to define the molecular basis for ?-amylase binding to the surface of the bacterium as well as the biological significance of this phenomenon in dental plaque biofilm formation. PMID:23144140
Sociodemographic Risk, Parenting, and Effortful Control: Relations to Salivary Alpha-amylase and Cortisol in Early Childhood
Taylor, Zoe E.; Spinrad, Tracy L.; VanSchyndel, Sarah K.; Eisenberg, Nancy; Huynh, Jacqueline; Sulik, Michael J.; Granger, Douglas A.
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
Sex Differences in Salivary Cortisol, Alpha-Amylase, and Psychological Functioning Following Hurricane Katrina
ERIC Educational Resources Information Center
Vigil, Jacob M.; Geary, David C.; Granger, Douglas A.; Flinn, Mark V.
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…
Studies on biological and enzymatic activities of salivary glands from the European hedgehog (Erinaceus europaeus).
Mebs, D
Aqueous extracts of salivary glands (Glandula submandibularis and G. parotis) from the European hedgehog (Erinaceus europaeus) exhibited neither lethal effect (intraperitoneal injection, mice), nor haemorrhagic and myonecrotic (mice) activity. Of the various enzymes tested (kallikrein, casein hydrolysis, phospholipase A2, acid and alkaline phosphatase, alpha-amylase), both glands possessed alkaline phosphatase and alpha-amylase activity only. These experiments suggest that toxic saliva in mammals is restricted to certain insectivores (shrews and solenodons) only. PMID:10482397
The relationship between cortisol, salivary alpha-amylase, and cognitive bias in young women.
Kreher, Donna A; Powers, Sally I; Granger, Douglas A
Both animal and human studies suggest that cognitive bias toward negative information, such as that observed in major depression, may arise through the interaction of cortisol (CORT) and norepinephrine (NE) within the amygdala. To date, there is no published account of the relationship between endogenous NE and CORT levels and cognitive bias. The present study examined salivary CORT and salivary alpha-amylase (sAA), an indirect measure of NE, in relation to masked affective priming of words in young female participants. Women with higher salivary CORT showed increased priming to negative word pairs only when sAA was also high; when sAA was low, no effect of CORT on priming was observed. These results are in line with previous research indicating that increased CORT is linked to enhanced processing of negative information. However, our findings extend this literature in providing evidence that CORT predicts enhanced processing of negatively valenced information only in the presence of higher sAA. PMID:22289045
Transglycosylation activity of Dictyoglomus thermophilum amylase A.
Nakajima, Masahiro; Imamura, Hiromi; Shoun, Hirofumi; Horinouchi, Sueharu; Wakagi, Takayoshi
Amylase A from Dictyoglomus thermophilum is a thermophilic enzyme and has about 40% identity with 4-alpha-glucanotransferase (GTase) from Thermococcus litoralis, and both of these enzymes belong to family 57 glycosyl hydrolase. Since the transglycosylation activity of T. litoralis GTase has been well characterized, the substrate specificity and reaction products of amylase A from D. thermophilum were examined. alpha-1,4 Glucan was produced from maltooligosaccharides, and glucoamylase-resistant molecules (cycloamyloses) were produced from longer chain amylose (average molecular mass 200 kDa). It has been reported that amylase A from D. thermophilum hydrolyzes starch, but in this study it was found that the enzyme was also able to use maltooligosaccharides and long chain amylose as substrate and has transglycosylation activity. PMID:15564678
Amylase activity is associated with AMY2B copy numbers in dog: implications for dog domestication, diet and diabetes
Arendt, Maja; Fall, Tove; Lindblad-Toh, Kerstin; Axelsson, Erik
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
Low copy number of the salivary amylase gene predisposes to obesity.
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
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
An in vitro and in vivo study of the ?-amylase activity of phaseolamin.
de Gouveia, Neire Moura; Alves, Fernanda Vieira; Furtado, Fabiana Barcelos; Scherer, Danielli Luana; Mundim, Antonio Vicente; Espindola, Foued Salmen
We evaluated the polypeptide profiles, inhibition of human salivary ?-amylase activity, and hemagglutination properties of a commercial phaseolamin sample. We also performed an in vivo assay to investigate the effects of a commercial phaseolamin treatment (100, 500, or 1500?mg/kg) over 20 days on the glycemia, body weight, and serum biochemical parameters (total cholesterol, triglycerides, alanine aminotransferase, and aspartate aminotransferase) of nondiabetic and streptozotocin-induced diabetic rats. The in vitro evaluation showed defined protein profiles, low hemagglutination activity, and high ?-amylase inhibition. None of the experimental groups treated with phaseolamin or acarbose showed decreases in body weight. Our data demonstrate that phaseolamin inhibits amylase activity in vitro, reduces blood glucose levels, decreases or attenuates some of the renal and hepatic effects of diabetes in streptozotocin-induced rats, and could therefore have therapeutic potential in the treatment or prevention of the complications of diabetes. PMID:24650210
Salivary-type amylase producing lung cancers examined clinically and pathologically in 260 Japanese patients of lung lobectomy or segmentectomy
Tadako Nakatsuji
The causative mechanisms of lung cancers producing salivary-type amylase (S-Amy) were hypothesized from clinical and pathological\\u000a analysis of lung cancers. A total of 260 Japanese patients who received lung lobectomy or segmentectomy during the last 4 years\\u000a at Hamamatsu University Hospital, Japan, were objective patients in this study. Among the 260 patients, 212 patients were\\u000a operated on for lung cancers. Of
Evening salivary alpha-amylase, major depressive disorder, and antidepressant use in the Netherlands Study of Depression and Anxiety (NESDA).
Veen, Gerthe; Giltay, Erik J; Licht, Carmilla M M; Vreeburg, Sophie A; Cobbaert, Christa M; Penninx, Brenda W J H; Zitman, Frans G
Salivary alpha-amylase (sAA) may be a suitable index for sympathetic activity and dysregulation of the autonomic nervous system. The relationship between antidepressants and depression with sAA levels was studied, since antidepressants were previously shown to have a profound impact on heart rate variability as an ANS indicator. Data are from 1692 participants of the Netherlands Study of Depression and Anxiety (NESDA) who were recruited from the community, general practice, and specialized mental health care. Differences in evening sAA levels were examined between patient groups (i.e., 752 current major depressive disorder [MDD], 611 remitted MDD, and 329 healthy controls) and between 46 tricyclic antidepressant (TCA) users, 307 selective serotonin reuptake inhibitor (SSRI) users, 97 users of another antidepressant, and 1242 non-users. Each participant sampled twice at 22.00h and 23.00h. In multivariable analysis, there was a trend over the three groups with increasing sAA levels from controls to remitted MDD to current MDD that approached significance. Furthermore, in comparison to non-users of antidepressants, TCA rather than SSRI users showed higher sAA levels, that persisted after multivariable adjustment. The present study shows that higher evening sAA levels in depressed patients, indicative of an increased sympathetic activity, may be induced by TCAs. PMID:23587658
The salivary alpha amylase over cortisol ratio as a marker to assess dysregulations of the stress systems.
Ali, Nida; Pruessner, Jens C
Different factors have been associated with changes in the regulation of the two major stress response systems of the human body, the sympathetic nervous system (SNS) and the hypothalamic-pituitary-adrenal (HPA) axis. Changes in these systems have been associated with various (psycho)pathologies across adulthood, and are thus frequently assessed within the context of allostatic load. Early Life Adversity (ELA) has been identified as one such factor. Individuals with histories of ELA show evidence of elevated basal and reactive salivary alpha amylase (sAA) levels (a marker of SNS activity), blunted cortisol levels (a marker of HPA axis activity), and an asymmetrical relationship between the two variables. However, variable methods used in the past to measure each variable, and the relationship between the two systems, prevent us from drawing firm conclusions. This preliminary study investigated whether the ratio of reactive sAA over reactive cortisol would be more informative to investigate the relationship between the two stress systems than the ratio of cortisol over sAA, or either marker alone, and whether there is a systematic link between this marker and subjective indexes of chronic stress and depression. We studied this in a total of 37 subjects (n=20 with signs of early life adversity and n=17 without) exposed to the Trier social stress test. Using a specific formula to determine the ratio of sAA over cortisol, we found a systematically stronger positive relationship with indexes of chronic stress and depression when compared to cortisol over sAA, or either marker alone. Our findings suggest that the ratio of sAA over cortisol might be a better marker of stress systems dysregulation than the ratio of cortisol over sAA, sAA or cortisol alone. The usefulness of this marker for other chronic stress states as found in allostatic load is discussed. PMID:22019784
The diurnal course of salivary alpha-amylase in nurses: an investigation of potential confounders and associations with stress.
Wingenfeld, Katja; Schulz, Michael; Damkroeger, Annika; Philippsen, Christine; Rose, Matthias; Driessen, Martin
In psychoneuroendocrinology research, salivary measures have become increasingly important. While several studies focus on determinants of salivary cortisol such as age, gender, and gynaecological variables, less research has focused on confounding variables of salivary alpha-amylase (sAA). In a large sample of nurses (N=215) we analyzed the impact of age, gender, intake of oral contraceptives, smoking, coffee consumption as well as psychological parameters, such as work stress and burnout, on basal diurnal sAA release. Saliva was collected at 07:00 h, 11:30 h, 17:30 h, and 20:00 h on a working day during early shift. Only gender could be identified to have an impact on sAA, with females having a more pronounced sAA increase over the course of the day. Whereas depression, anxiety, work stress and burnout were not associated with sAA, a small negative correlation between social difficulties, measured with the Chronic Stress Screening Scale, and sAA could be identified. PMID:20433894
Activity and cellular localization of amylases of rabbit cecal bacteria.
Sirotek, K; Marounek, M; Suchorská, O
Five 11-week-old rabbits, fed a commercial granulated feed, were slaughtered and cecal starch-degrading bacteria enumerated; total concentration of cultivable bacteria utilizing starch averaged 5.5 x 10(10) CFU/g. The activity and cellular localization of amylases was determined in 9 bacteria identified as Actinomyces israeli (strains AA2 and AD4), Bacteroides spp. (strain AA3), Dichelobacter nodosus (strain AA4), Mitsuokella multiacidus (strain AA6), Eubacterium spp. (strains AA7 and AB2), Clostridium spp. (strains AD1 and AA5). Four strains (AA3, AA4, AA5, AD4) produced extracellular amylases with an activity of 26-35 micromol of reducing sugars per h per mg of protein; in five strains (AA2, AA6, AA7, AB2, AD1) amylases were membrane-bound with an activity of 14-18 micromol of reducing sugars per h per mg of protein. All strains exhibited a low intracellular amylolytic activity. The pH optimum of amylases was 6.8-7.0. In strains producing extracellular amylases a substantial loss of viscosity was observed during incubations of cultivation supernatant with starch, similar to viscosity reduction in starch solutions treated with alpha-amylase; this indicates an endo-type (random cleavage) of extracellular amylase reaction in the bacteria under study. No strain possessed glucoamylase activity. PMID:17007433
Asymmetry in children's salivary cortisol and alpha-amylase in the context of marital conflict: links to children's emotional security and adjustment.
Koss, Kalsea J; George, Melissa R W; Cummings, E Mark; Davies, Patrick T; El-Sheikh, Mona; Cicchetti, Dante
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
Activity and storage of commercial amylases in the 2013 Louisiana grinding season
Technology Transfer Automated Retrieval System (TEKTRAN)
A current problem in the application of amylases at sugarcane factories is the existence of a wide variation in the activities and activity per unit cost of commercial amylases. The efficiency of amylase action to break down starch in the factory is related to the activity of the amylase used. Until...
Pouteria ramiflora extract inhibits salivary amylolytic activity and decreases glycemic level in mice.
De Gouveia, Neire M; De Albuquerque, Cibele L; Espindola, Laila S; Espindola, Foued S
In this study, extracts of plant species from the Cerrado biome were assessed in order to find potential inhibitors of human salivary alpha-amylase. The plants were collected and extracts were obtained from leaves, bark, and roots. We performed a preliminary phytochemical analysis and a screening for salivar alpha-amylase inhibitory activity. Only three botanical families (Sapotaceae, Sapindaceae and Flacourtiaceae) and 16 extracts showed a substantial inhibition (>75%) of alpha-amylase. The ethanolic extracts of Pouteria ramiflora obtained from stem barks and root barks decreased amylolytic activity above 95% at a final concentration of 20 µg/mL. Thus, adult male Swiss mice were treated orally with P. ramiflora in acute toxicity and glycemic control studies. Daily administration with 25, 50 and 100 mg/kg of aqueous extract of P. ramiflora for eight days can reduce significantly body weight and blood glucose level in mice. These data suggest that the crude polar extract of P. ramiflora decreases salivary amylolytic activity while lowering the blood levels of glucose. PMID:24068095
Fractionated irradiation and early changes in salivary glands. Different effects on potassium efflux, exocytotic amylase release and gland morphology
Franzen, L.; Funegard, U.S.; Sundstroem, S.G.; Gustafsson, H.; Danielsson, A.; Henriksson, R. (University Hospital, Umea (Sweden))
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.
Interactions between salivary cortisol and alpha-amylase as predictors of children's cognitive functioning and academic performance.
Keller, Peggy S; El-Sheikh, Mona; Granger, Douglas A; Buckhalt, Joseph A
We examined relations between salivary cortisol, alpha-amylase (sAA), and children's cognitive and academic functioning. Of interest were curvilinear and interactive effects of these salivary measures on cognitive and academic performance. Data were based on a sample of 28 boys and 36 girls (ages 8 and 9) in the Southeastern U.S.A. Children provided resting afternoon saliva samples. Children completed standardized tests of Intellectual Ability and schools provided academic achievement information. Regression analyses demonstrated significant curvilinear relations and interactions between cortisol and sAA in the prediction of child functioning. Contrary to current models of interactions among biological systems, findings indicated some of the highest and lowest scores were predicted at moderate levels of physiological arousal. For example, children with moderate sAA and either higher or lower cortisol had low predicted scores for Reading Ability. Children with moderate cortisol and lower sAA had the highest predicted scores for Intellectual Ability. Findings suggest that the study of interactions between biological stress response systems should not be based on models of rectilinear interactions. PMID:22100627
Crystallization and preliminary X-ray diffraction studies of human salivary alpha-amylase.
Ramasubbu, N; Bhandary, K K; Scannapieco, F A; Levine, M J
Nonglycosylated alpha-amylase, a major component of human parotid saliva, has been crystallized by the vapor diffusion technique using 2-methyl-2,4-pentanediol as the precipitant in the presence of CaCl2 at pH 9.0. The crystals are orthorhombic, space group P2(1)2(1)2(1) with unit cell dimensions of a = 53.3, b = 75.8, and c = 138.1 A. The asymmetric unit contains one amylase molecule. The solvent content is 54%. The crystals are stable to X-rays and diffract up to 2.8 A and appear to be suitable for X-ray diffraction studies. PMID:1749776
Intracellular ?-Amylase of Streptococcus mutans
Simpson, Christine L.; Russell, Roy R. B.
Sequencing upstream of the Streptococcus mutans gene for a CcpA gene homolog, regM, revealed an open reading frame, named amy, with homology to genes encoding ?-amylases. The deduced amino acid sequence showed a strong similarity (60% amino acid identity) to the intracellular ?-amylase of Streptococcus bovis and, in common with this enzyme, lacked a signal sequence. Amylase activity was found only in S. mutans cell extracts, with no activity detected in culture supernatants. Inactivation of amy by insertion of an antibiotic resistance marker confirmed that S. mutans has a single ?-amylase activity. The amylase activity was induced by maltose but not by starch, and no acid was produced from starch. S. mutans can, however, transport limit dextrins and maltooligosaccharides generated by salivary amylase, but inactivation of amy did not affect growth on these substrates or acid production. The amylase digested the glycogen-like intracellular polysaccharide (IPS) purified from S. mutans, but the amy mutant was able to digest and produce acid from IPS; thus, amylase does not appear to be essential for IPS breakdown. However, when grown on excess maltose, the amy mutant produced nearly threefold the amount of IPS produced by the parent strain. The role of Amy has not been established, but Amy appears to be important in the accumulation of IPS in S. mutans grown on maltose. PMID:9721315
Who is stressed? A pilot study of salivary cortisol and alpha-amylase concentrations in agoraphobic patients and their novice therapists undergoing in vivo exposure.
Schumacher, Sarah; Gaudlitz, Katharina; Plag, Jens; Miller, Robert; Kirschbaum, Clemens; Fehm, Lydia; Fydrich, Thomas; Ströhle, Andreas
In cognitive behavioural therapy of phobic anxiety, in vivo exposure is considered as an effective treatment strategy. Apparently, it involves the experience of stress and anxiety in patients. Given the therapist's role during exposure sessions, it is conceivable that the performance is also accompanied with the experience of stress in therapists, especially when unversed in conducting psychotherapy. Studies confirmed that cognitive behavioural therapists tend to avoid therapist-guided in vivo exposure. The objective of this study was the simultaneous investigation of therapist's and patient's stress response during in vivo exposure. Therefore, 23 agoraphobic patients and their 23 treating therapists in training provided five saliva samples during an in vivo exposure and five samples during an ordinary therapy session. Before and during exposure session, subjective evaluations of stress and anxiety were assessed. Results suggested that therapists reported similar levels of perceived stress as patients before exposure. Both groups displayed significantly elevated salivary cortisol (sC) levels during exposure compared to the control session and a trend for alterations in salivary alpha-amylase (sAA) activity was found. Therapists reached peak concentrations of sC before start of the intervention followed by a decline during exposure, while patients displayed peak levels of cortisol secretion after 60 min of exposure. In vivo exposure seems to be a demanding intervention not only for the patient, but also for therapists in training. However, it was also demonstrated that physiological and subjective stress rather decrease during the intervention and that both groups rated exposure to be substantially successful. Based on the presented results, another potential factor contributing to the under-usage of exposure treatment is conceivable and needs to be addressed in future research. PMID:25127086
Effect of Chronic Training on Heart Rate Variability, Salivary IgA and Salivary Alpha-Amylase in Elite Swimmers with a Disability
Edmonds, Rohan
The purpose of this study was to a) determine the heart rate variability (HRV) and saliva markers of immunity (salivary immunoglobulin A; sIgA) and stress (salivary alpha-amylase; sAA) responses to chronic training in elite swimmers with a disability; and b) identify the relationships between HRV, sIgA, sAA and training volume. Eight members of a high performance Paralympic swimming program were monitored for their weekly resting HRV, sIgA and sAA levels in the 14 weeks leading up to a major international competition. The 14 week training program included aerobic, anaerobic, power and speed, and taper training phases, while also incorporating two swimming step tests and two swimming competitions. Specific time (root mean square of the successive differences; RMSSD) and frequency (high frequency normalized units [HFnu]) domain measures, along with non-linear indices (standard deviation of instantaneous RR variability; SD1 and short term fractal scaling exponent; ?1) of HRV were used for all analyses with effects examined using magnitude-based inferences. Relationships between HRV and saliva markers were identified by Spearman rank rho (?) correlation coefficients. Compared with week 1, SD1 was very likely lower (96/4/0, ES = -2.21), while sAA was very likely elevated (100/0/0, ES = 2.32) at the beginning of week 7 for all athletes. The training program did not alter HRV or saliva whereas competition did. There were also no apparent differences observed for HRV, sIgA and sAA between each of the training phases during the 14 week swimming program. Correlations were observed between sAA and SD1 (? = -0.212, p<0.05), along with sAA and mean HR (? = 0.309, p<0.05). These results show that high level national competition influences depresses HRV (SD1) and increases saliva biomarkers of stress (sAA). It appears that a well-managed and periodised swimming program can maintain these indices within normal baseline levels. The study also highlighted the parasympathetic nervous system influence on sAA. PMID:26043224
Other nonstress influences can alter salivary ?-amylase activity.
Stegmann, Barbara Jean
Clearly, more research is required to fully evaluate the impact of stress on both fertile and infertile women, and discovery of a biomarker that correlates well with psychosocial stress would be a great advantage to researchers. This article is a first step toward that goal, but it may be premature to assign these findings to stress alone. PMID:21601665
Halotolerant Ability and ?-Amylase Activity of Some Saltwater Fungal Isolates.
Niknejad, Farhad; Moshfegh, Mahsa; Najafzadeh, Mohammad Javad; Houbraken, Jos; Rezaei, Shahla; Zarrini, Gholamreza; Faramarzi, Mohammad Ali; Nafissi-Varcheh, Nastaran
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
Halotolerant Ability and ?-Amylase Activity of Some Saltwater Fungal Isolates
Niknejad, Farhad; Moshfegh, Mahsa; Najafzadeh, Mohammad Javad; Houbraken, Jos; Rezaei, Shahla; Zarrini, Gholamreza; Faramarzi, Mohammad Ali; Nafissi-Varcheh, Nastaran
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
Salivary alpha-amylase during pregnancy: diurnal course and associations with obstetric history, maternal demographics, and mood.
Giesbrecht, Gerald F; Granger, Douglas A; Campbell, Tavis; Kaplan, Bonnie
Diurnal patterns of salivary alpha amylase (sAA) in pregnant women have not previously been described. The current study employed ecological momentary assessment to examine the association between the diurnal sAA, obstetric history, maternal demographics, and mood during pregnancy. Saliva was self-collected by 83 pregnant women (89% White, age 25.3-43.0 years; mean gestational age 21.9 weeks, range 6-37 weeks; gravida 1-6) at home over three days. Results indicated that current pregnancy (gestational age and fetal sex) and maternal demographics were not related to diurnal sAA. In contrast, a history of previous miscarriage (Parameter = -.17; SE = .05; p < .05) was associated with an atypical diurnal pattern. Even after accounting for obstetric history, trait anxiety (Parameter = .16; SE = .04; p < .001) was associated with increased sAA over the day while chronic levels of fatigue (Parameter = -.06; SE = .03; p < .05) were associated with decreased sAA. In a separate model, we also tested the time varying covariation of sAA and mood. The effects of momentary mood were in contrast to those for trait mood. Both momentary depression (Parameter = .22; SE = .09; p < .01) and vigour/positive mood (Parameter = .12; SE = .04; p < .001) were associated with momentary increases in sAA while momentary anxiety and fatigue were not related to sAA. The findings suggest that basal sAA during pregnancy is sensitive to emotional arousal. Evaluating diurnal patterns of sAA holds promise for advancing understanding of how emotional arousal during pregnancy may affect fetal development. PMID:22315130
Effects of Hatha Yoga on Blood Pressure, Salivary ?-Amylase, and Cortisol Function Among Normotensive and Prehypertensive Youth
Mueller, Martina; Gregoski, Mathew J.; Brunner-Jackson, Brenda; McQuade, Lisa; Matthews, Cameron; Treiber, Frank A.
Abstract Objective: Evidence is accumulating, predominantly among clinical trials in adults, that yoga improves blood pressure (BP) control, with downregulation of the hypothalamic–pituitary–adrenal (HPA) axis and the sympathetic nervous system (SNS) projected as underlying mechanisms. This pilot study assessed whether Hatha yoga has the potential to reduce BP among youth and whether dampening of the SNS and/or HPA activity is a likely pathway of change. Design: Thirty-one seventh graders were randomly assigned to a Hatha yoga program (HYP) or attention control (AC) music or art class. Baseline and 3-month evaluations included resting BP; overnight urine samples; and saliva collected at bedtime, upon awakening, and at 30 and 60 minutes after awakening for ?-amylase and cortisol assays. Results: Twenty-eight (14 in the HYP group and 14 in the AC group) students were assessed both before and after the intervention. BP changes from pre- to post-intervention were ?3.0/?2.0?mmHg for the HYP group and ?0.07/?0.79?mmHg for the AC group (p=0.30 and 0.57, respectively). Changes in systolic BP (SBP)/diastolic BP (DBP) for the prehypertensive (75th–94th percentiles for SBP) subgroup analyses were ?10.75/?8.25?mmHg for the HYP group (n=4) versus 1.8/1.0?mmHg for the AC group (n=5) (p for SBP=0.02; p for DBP=0.09). Although no statistically significant group differences were observed with changes in SNS or HPA awakening curves (area under curve for ?-amylase and cortisol, respectively), a small to moderate effect size was seen favoring a reduction of ?-amylase activation for the HYP group (Cohen d=0.34; prehypertensive d=0.20). Conclusions: A school-based Hatha yoga program demonstrated potential to decrease resting BP, particularly among prehypertensive youth. Reduced SNS drive may be an underlying neurohormonal pathway beneficially affected by the program. A large-scale efficacy/effectiveness randomized clinical trial is warranted. PMID:24620850
Passive sorting in maturing granules of AtT-20 cells: the entry and exit of salivary amylase and proline-rich protein.
Castle, A M; Huang, A Y; Castle, J D
Previous studies have suggested that salivary amylase and proline-rich protein are sorted differently when expressed in AtT-20 cells (Castle, A.M., L.E. Stahl, and J.D. Castle. 1992. J. Biol. Chem. 267:13093- 13100; Colomer, V., K. Lal, T.C. Hoops, and M.J. Rindler. 1994.EMBO (Eur. Mol. Biol. Organ.) J. 13:3711- 3719). We now show that both exocrine proteins behave similarly and enter the regulated secretory pathway as judged by immunolocalization and secretagogue- dependent stimulation of secretion. Analysis of stimulated secretion of newly synthesized proline-rich protein, amylase, and endogenous hormones indicates that the exogenous proteins enter the granule pool with about the same efficiency as the endogenous hormones. However, in contrast to the endogenous hormones, proline-rich protein and amylase are progressively removed from the granule pool during the process of granule maturation such that only small portions remain in mature granules where they colocalize with the stored hormones. The exogenous proteins that are not stored are recovered from the incubation medium and are presumed to have undergone constitutive-like secretion. These results point to a level of sorting for regulated secretion after entry of proteins into forming granules and indicate that retention is essential for efficient storage. Consequently, the critical role of putative sorting receptors for regulated secretion may be in retention rather than in granule entry. PMID:9214380
Influence of tableting on the enzymatic activity of different ?-amylases using various excipients
Katharina M Picker
The purpose of the study was to show the influence of compression pressure on the enzymatic activity of different types of ?-amylases and to analyze the loss of activity of ?-amylase in mixtures with different excipients. Following that, the properties of excipients used for tableting enzymes were evaluated. Tablets were produced on an instrumented single punch tableting machine. The pure
Influence of local air velocity from air conditioner evaluated by salivary and skin biomarkers
Masaki Yamaguchi; Takayuki Takahashi; Yuichiro Yoshino; Makoto Sasaki; Hajime Nishimiya
The purpose of this paper is to reveal both the psychosomatic and the physical effects of local air velocity from an air conditioner using biomarkers which can be collected noninvasively. Salivary ?-amylase activity (SAA) and salivary cortisol were used as the indexes of psychosomatic effects. The total protein (TP) collected from stratum corneum was used as an index of the
Vasodilatory activity in horsefly and deerfly salivary glands.
Rajská, P; Pechánová, O; Takác, P; Kazimírová, M; Roller, L; Vidlicka, L; Ciampor, F; Labuda, M; Nuttall, P A
Salivary gland extract (SGE) of four horsefly species (Hybomitra bimaculata Macquart, Hybomitra ciureai Séguy, Tabanus bromius L., Tabanus glaucopis Meigen) and one deerfly species (Chrysops relictus Meigen) (Diptera: Tabanidae) were shown to contain vasodilatory activity. Aliquots equivalent to 1, 5 and 10 pairs of salivary glands (SG) relaxed rat femoral artery (with intact endothelium) pre-constricted with phenylephrine. Vasodilatory activity was dose-dependent. SGE of one horsefly species (Haematopota pluvialis L.) did not induce relaxation. The kinetics of vasodilation induced by SGE of four horsefly species differed from the deerfly. These results indicate that tabanid species may produce more than one type of vasodilator to aid blood feeding. PMID:14651653
Characteristic hydrolyzing of megalosaccharide by human salivary ?-amylase and small intestinal enzymes, and its bioavailability in healthy subjects.
Nakamura, Sadako; Takami, Masayuki; Tanabe, Kenichi; Oku, Tsuneyuki
The digestibility of Megalosaccharide® (newly developed carbohydrate comprising ?-1,4-glucosaccharide) was investigated in vitro and in vivo. Isomaltosyl-megalosaccharide® (IMS) and nigerosyl-megalosaccharide® (NMS) contain 20% and 50% of the megalosaccharide fraction (degree of polymerization (DP) 10-35), respectively. IMS was hydrolyzed readily by ?-amylase to oligosaccharides (DP???7), and a small amount of glucose was produced from oligosaccharides by small intestinal enzymes (SIEs). NMS was partially hydrolyzed by ?-amylase to oligosaccharides, and a small amount of glucose produced by SIEs. When IMS and NMS were treated by SIEs after treatment with human saliva ?-amylase for a few minutes, IMS and NMS were hydrolyzed readily to glucose. Plasma levels of glucose and insulin upon ingestion of 50?g of IMS or NMS were elevated the same as those for 50?g of glucose, and breath hydrogen was not excreted. These results suggest that IMS and NMS are digestible carbohydrates. PMID:24725210
Potentiometric flow injection determination of amylase activity by using hexacyanoferrate(III)-hexacyanoferrate(II) potential buffer
Hiroki Ohura; Toshihiko Imato; Yasukazu Asano; Sumio Yamasaki
A highly sensitive potentiometric flow injection determination of amylase activity was carried out, utilizing a redox reaction of hexacyanoferrate(III) in alkaline media with reducing sugar as product of the enzymatic hydrolysis reaction of starch with amylase. The analytical method is based on the potential change detection of a flow-through type redox electrode detector due to the composition change of a
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...
Studies on the Utility of B-Amylase1 IntronIII Sequences as Markers for B-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 in...
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...
Proteolytic activity of Candida albicans: action on human salivary proteins.
Germaine, G R; Tellefson, L M; Johnson, G L
The susceptibility of human salivary proteins to degradation by Candida albicans was studied. The organism was cultivated in either whole-salivary supernatant or parotid fluid, both of which were supplemented with glucose (0.1%). The culture pH's were at, or above, neutrality. After growth, the culture supernatant solutions were examined by polyacrylamide gel electrophoresis for alterations in their profiles of salivary proteins. No evidence of proteolysis of whole-saliva or parotid fluid proteins was found. Salivary proteins, however, are susceptible to degradation by preparations of C. albicans protease. Candida protease was incubated with parotid fluid adjusted to several pH values. After incubation the reaction mixtures were subjected to polyacrylamide gel electrophoresis. Extensive degradation of parotid proteins was found at pH 4, very slight proteolysis at pH 5, and no degradation at pH 6 or 7. No selectivity in proteolysis of the several parotid proteins was noted. These results indicate that C. albicans protease is strictly dependent upon low (ca. 4) pH for activity on salivary proteins. Furthermore, it is suggested that due to the pH requirements of the enzyme, it is unlikely to be of major significance to the pathogenesis of Candida-induced oral inflammatory lesions. PMID:32141
Production of alpha-amylase by yeast
Thomse, K.K.
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).
[Amylase-producing multiple myeloma: a case report].
Mori, Minako; Maruoka, Hayato; Nagai, Yuya; Fujita, Haruhiko; Togami, Katsuhiro; Tabata, Sumie; Kurata, Masahiro; Matsushita, Akiko; Nagai, Kenichi; Tanaka, Kyoko; Yamashiro, Akiko; Takahashi, Takayuki
A 80-year-old man was admitted because of acute-onset thrombocytopenia and renal failure. He was diagnosed with Bence Jones (lambda) -type multiple myeloma associated with sepsis with methicillin-resistant Staphylococcus aureus. On admission, serum amylase activity was elevated to 1,814 IU/l (98% salivary type; S-amylase). Several days after admission, he developed bilateral myelomatous pleuritis. The activity of S-amylase in the effusion was 5,495 IU/l. Myeloma cells in the pleural effusion were positive for cytoplasmic amylase with an antibody against human amylase. High S-amylase activity was detected in the supernatant of cultured myeloma cells in the effusion. Furthermore, S-amylase gene expression was detected by RT-PCR. A diagnosis of amylase-producing multiple myeloma was made. The patient died of renal insufficiency complicated by severe DIC. We report a rare case of amylase-producing myeloma confirmed by immunocytochemistry, culture method, and gene expression. PMID:18080506
Adrenoceptor-activated Nitric Oxide Synthesis in Salivary Acinar Cells
D. K. Looms; S. Dissing; K. Tritsaris; A. M. Pedersen; B. Nauntofte
We investigated the cellular regulation of nitric oxide synthase (NOS) activity in isolated acinar cells from rat parotid and human labial salivary glands, using the newly developed fluorescent nitric oxide (NO) indicator, DAF-2. We found that sympathetic stimulation with norepinephrine (NE) caused a strong increase in NO synthesis that was not seen after parasympathetic stimulation with acetylcholine. In rat parotid
Alpha-Amylase Activity in Blood Increases after Pharmacological, But Not Psychological, Activation of the Adrenergic System
Nater, Urs M.; La Marca, Roberto; Erni, Katja; Ehlert, Ulrike
Background & Aim Alpha-amylase in both blood and saliva has been used as a diagnostic parameter. While studies examining alpha-amylase activity in saliva have shown that it is sensitive to physiological and psychological challenge of the adrenergic system, no challenge studies have attempted to elucidate the role of the adrenergic system in alpha-amylase activity in blood. We set out to examine the impact of psychological and pharmacological challenge on alpha-amylase in blood in two separate studies. Methods In study 1, healthy subjects were examined in a placebo-controlled, double-blind paradigm using yohimbine, an alpha2-adrenergic antagonist. In study 2, subjects were examined in a standardized rest-controlled psychosocial stress protocol. Alpha-amylase activity in blood was repeatedly measured in both studies. Results Results of study 1 showed that alpha-amylase in blood is subject to stronger increases after injection of yohimbine compared to placebo. In study 2, results showed that there was no significant effect of psychological stress compared to rest. Conclusions Alpha-amylase in blood increases after pharmacological activation of the adrenergic pathways suggesting that sympathetic receptors are responsible for these changes. Psychological stress, however, does not seem to have an impact on alpha-amylase in blood. Our findings provide insight into the mechanisms underlying activity changes in alpha-amylase in blood in healthy individuals. PMID:26110636
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...
An analytical method for measuring ?-amylase activity in starch-containing foods.
Koyama, Kazuo; Hirao, Takashi; Toriba, Akira; Hayakawa, Kazuichi
The quality of starch-containing foods may be significantly impaired by contamination with very small amounts of ?-amylase, which can enzymatically hydrolyze the starch and cause viscosity loss. Thus, for quality control, it is necessary to have an analytical method that can measure low amylase activity. We developed a sensitive analytical method for measuring the activity of ?-amylase (from Bacillus subtilis) in starch-containing foods. The method consists of six steps: (1) crude extraction of ?-amylase by centrifugation and filtration; (2) ?-amylase purification by desalting and anion-exchange chromatography; (3) reaction of the purified amylase with boron-dipyrromethene (BODIPY)-labeled substrate, which releases a fluorescent fragment upon digestion of the substrate, thus avoiding interference from starch derivatives in the sample; (4) stopping the reaction with acetonitrile; (5) reversed-phase solid-phase extraction of the fluorescent substrate to remove contaminating dye and impurities; and (6) separation and measurement of BODIPY fluorescence by HPLC. The proposed method could quantify ?-amylase activities as low as 10 mU/mL, which is enough to reduce the viscosity of starch-containing foods. PMID:23074083
Inhibition of ?-Amylase and ?-Glucosidase Activity by Tea and Grape Seed Extracts and their Constituent Catechins
Yilmazer-Musa, Meltem; Griffith, Anneke M.; Michels, Alexander J.; Schneider, Erik; Frei, Balz
We evaluated the inhibitory effects of plant-based extracts (grape seed, green tea, and white tea) on ?-amylase and ?-glucosidase activity, glucosidases required for starch digestion. The abundant flavan-3-ol monomers (catechins) in these extracts were also tested for their inhibitory potential and evaluated against the pharmacological glucosidase inhibitor, acarbose. To evaluate relative potency of these extracts and catechins, the concentrations required for 50 and 90% inhibition of enzyme activity were determined. Maximum enzyme inhibition was used to assess an inhibitor's relative efficacy. Results showed that grape seed extract strongly inhibited both ?-amylase and ?-glucosidase activity, with equal and much higher potency, respectively, than acarbose. While tea extracts and individual catechin 3-gallates were less effective inhibitors of ?-amylase, they were potent inhibitors of ?-glucosidase. Our data show that plant extracts containing catechin 3-gallates are potent inhibitors of ?-glucosidase, and suggest that procyanidins found in grape seed extract strongly inhibit ?-amylase activity. PMID:22697360
Amylase-Binding Protein B of Streptococcus gordonii Is an Extracellular Dipeptidyl-Peptidase?
Chaudhuri, Biswendu; Paju, Susanna; Haase, Elaine M.; Vickerman, M. Margaret; Tanzer, Jason M.; Scannapieco, Frank A.
The oral commensal bacterium Streptococcus gordonii interacts with salivary amylase via two amylase-binding proteins, AbpA and AbpB. Based on sequence analysis, the 20-kDa AbpA protein is unique to S. gordonii, whereas the 82-kDa AbpB protein appears to share sequence homology with other bacterial dipeptidases. The aim of this study was to verify the peptidase activity of AbpB and further explore its potential functions. The abpB gene was cloned, and histidine-tagged AbpB (His-AbpB) was expressed in Escherichia coli and purified. Its amylase-binding activity was verified in an amylase ligand binding assay, and its cross-reactivity was verified with an anti-AbpB antibody. Both recombinant His-AbpB and partially purified native AbpB displayed dipeptidase activity and degraded human type VI collagen and fibrinogen, but not salivary amylase. Salivary amylase precipitates not only AbpA and AbpB but also glucosyltransferase G (Gtf-G) from S. gordonii supernatants. Since Streptococcus mutans also releases Gtf enzymes that could also be involved in multispecies plaque interactions, the effect of S. gordonii AbpB on S. mutans Gtf-B activity was also tested. Salivary amylase and/or His-AbpB caused a 1.4- to 2-fold increase of S. mutans Gtf-B sucrase activity and a 3- to 6-fold increase in transferase activity. An enzyme-linked immunosorbent assay verified the interaction of His-AbpB and amylase with Gtf-B. In summary, AbpB demonstrates proteolytic activity and interacts with and modulates Gtf activity. These activities may help explain the crucial role AbpB appears to play in S. gordonii oral colonization. PMID:18678669
Amylase-binding protein B of Streptococcus gordonii is an extracellular dipeptidyl-peptidase.
Chaudhuri, Biswendu; Paju, Susanna; Haase, Elaine M; Vickerman, M Margaret; Tanzer, Jason M; Scannapieco, Frank A
The oral commensal bacterium Streptococcus gordonii interacts with salivary amylase via two amylase-binding proteins, AbpA and AbpB. Based on sequence analysis, the 20-kDa AbpA protein is unique to S. gordonii, whereas the 82-kDa AbpB protein appears to share sequence homology with other bacterial dipeptidases. The aim of this study was to verify the peptidase activity of AbpB and further explore its potential functions. The abpB gene was cloned, and histidine-tagged AbpB (His-AbpB) was expressed in Escherichia coli and purified. Its amylase-binding activity was verified in an amylase ligand binding assay, and its cross-reactivity was verified with an anti-AbpB antibody. Both recombinant His-AbpB and partially purified native AbpB displayed dipeptidase activity and degraded human type VI collagen and fibrinogen, but not salivary amylase. Salivary amylase precipitates not only AbpA and AbpB but also glucosyltransferase G (Gtf-G) from S. gordonii supernatants. Since Streptococcus mutans also releases Gtf enzymes that could also be involved in multispecies plaque interactions, the effect of S. gordonii AbpB on S. mutans Gtf-B activity was also tested. Salivary amylase and/or His-AbpB caused a 1.4- to 2-fold increase of S. mutans Gtf-B sucrase activity and a 3- to 6-fold increase in transferase activity. An enzyme-linked immunosorbent assay verified the interaction of His-AbpB and amylase with Gtf-B. In summary, AbpB demonstrates proteolytic activity and interacts with and modulates Gtf activity. These activities may help explain the crucial role AbpB appears to play in S. gordonii oral colonization. PMID:18678669
Human Salivary Mucin MG1 Selectively Forms Heterotypic Complexes with Amylase, Proline-rich Proteins, Statherin, and Histatins
I. Iontcheva; F. G. Oppenheim; R. F. Troxler
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,
Emergence in human dental plaque and host distribution of amylase-binding streptococci.
Scannapieco, F A; Solomon, L; Wadenya, R O
Salivary amylase is known to bind specifically to several species of oral streptococci. To assess the importance of this interaction in bacterial colonization of the oral cavity, we determined the proportion and identity of amylase-binding bacteria (ABB) in dental plaque of humans and various salivary amylase-secreting and non-secreting mammalian species. The numbers of ABB in undisturbed plaque collected over time from tooth surfaces of six human volunteers or from 14 other mammalian species were determined by means of a replicating assay. The mean proportion of ABB cultured aerobically from human teeth at 2 h was 10.5% (SD 10), at 8 h 7.9% (8), at 24 h 13% (11), and at 48 h 12% (9). The mean proportion of anaerobically cultured ABB found at 2 h was 3% (SD 4), at 8 h 5% (5), at 24 h 12% (9), and at 48 h 16% (12). Amylase-binding bacteria cultured from these samples resembled Streptococcus mitis, Streptococcus gordonii, Streptococcus salivarius, Streptococcus crista, or unidentified streptococci. In addition, only animals exhibiting salivary amylase activity in their saliva harbored ABB (ranging from 2 to 31% of the total flora), with the exception of the pig, where no ABB were found to colonize, despite considerable amylase activity in saliva. Only strains resembling S. mitis and S. salivarius and unspeciated strains were isolated from these mammals. These results suggest that amylase-binding streptococci are the predominant ABB in human plaque, and their numbers generally increase as plaque develops. Since ABB colonized only the oral cavities of hosts demonstrating salivary amylase activity, the ability to bind amylase may play an important role in oral colonization by these bacteria. PMID:7523468
LEADER 3—Lipase and Amylase Activity in Subjects With Type 2 Diabetes
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.
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
Salivary steroid assays for assessing variation in endocrine activity.
Riad-Fahmy, D; Read, G F; Walker, R F
Salivary sampling regimens are non-invasive, and therefore facilitate dynamic tests of hormone function and assessment of biological rhythms. Concentrations of neutral steroids in saliva are independent of flow rate and appear to reflect the non-protein-bound, 'free' fraction. Comparison of replicate determinations of quality control pools with determinations of samples collected at 2 min intervals allows the significance of short-term fluctuations in cortisol and testosterone secretory activity to be estimated. Samples collected at 15 min intervals provide a convenient way to estimate circadian rhythms, particularly in young children. Determination of salivary progesterone concentrations in samples collected by women daily, over extended periods of time, provides a valuable means of assessing ovarian function. Such assays may be used to monitor ovulation-induction therapy. PMID:6887863
The effect of chronic treatment with fluoride on salivary activity, tooth, and bone in spontaneously hypertensive rats (SHR).
Picco, Daniele C R; Delbem, Alberto C B; Sassaki, Kikue T; Sumida, Doris H; Antoniali, Cristina
The present study evaluated the effect of chronic treatment with sodium fluoride on salivary activity, tooth, and bone in spontaneously hypertensive rats (SHR). The treatment was made with a 20-ppm NaF solution added to the drinking water for 30 days. Systolic blood pressure values were obtained by plethysmography; fluoride concentration was determined by an ion-selective electrode; calcium concentration and amylase activity were determined by commercial kits; and enamel microhardness was verified by longitudinal section. Systolic blood pressure values and animals' weight were not changed by treatment. However, the salivary flow rate-which was lowered in SHR at baseline when compared to Wistar rats-was found to be increased with the treatment with NaF. The fluoride concentration was increased in the plasma of the treated groups, even though it remained lower for the treated SHR in relation to the treated Wistar rats. Calcium concentration was decreased in the saliva and plasma of SHR treated with NaF. A reduction in the plasmatic total protein concentration was observed in SHR treated with NaF. The fluoride concentration on bone surface was found to be increased in Wistar or SHR treated with NaF. In treated SHR's femurs, it was observed a significant reduction in fluoride concentrations. Enamel microhardness of the incisor teeth was not changed by the treatment with NaF in both groups. The distribution of fluoride to the salivary glands in SHR is poor, and treatment with NaF causes a decrease in the concentration of important biochemical parameters to the salivary physiology in SHR. PMID:24390229
Immobilization of ?-amylase onto a calix[4]arene derivative: Evaluation of its enzymatic activity.
Veesar, Irshad Ali; Solangi, Imam Bakhsh; Memon, Shahabuddin
In order to enhance the cost-effectiveness practicability of enzymes in many industries such as pharmaceutical, food, medical and some other technological processes, there is great need to immobilize them onto a solid supports. In this study, a new and efficient immobilization of ?-amylase from Saccharomyces cerevisiae has been developed by using the surface functionalization of calix[4]arene as support. A glutaraldehyde-containing amino group functionalized calix[4]arene was used to immobilize ?-amylase covalently. In this procedure, imide bonds are formed between amino groups on the protein and aldehyde groups on the calix[4]arene surface. The surface modified support was characterized using Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM). The effect of various preparation conditions on the immobilized ?-amylase process such as immobilization time, enzyme concentration, temperature and pH were investigated. The influence of pH and temperature on the activity of free and immobilized ?-amylase was also studied using starch as substrate. The optimum reaction temperature and pH value for the enzymatic conversion catalyzed by the immobilized ?-amylase were 25°C and 7, respectively. Compared to the free enzyme, the immobilized ?-amylase retained 85% of its original activity and exhibited significant thermal stability than the free one and excellent durability. PMID:25965976
RESEARCH ARTICLE Open Access Caffeine administration does not alter salivary
Ritter, Frank
RESEARCH ARTICLE Open Access Caffeine administration does not alter salivary -amylase activity in young male daily caffeine consumers Laura Cousino Klein1,2* , Courtney A Whetzel1 , Jeanette M Bennett1 report from our lab [Hum Psychopharmacol 25:359367, 2010.] we examined the effects of caffeine
Potent ?-amylase inhibitory activity of Indian Ayurvedic medicinal plants
Sudha P; Smita S Zinjarde; Shobha Y Bhargava; Ameeta R Kumar
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
Salivary Acetylcholinesterase Activity Is Increased in Parkinson's Disease: A Potential Marker of Parasympathetic Dysfunction
Fedorova, Tatyana; Knudsen, Cindy Soendersoe; Mouridsen, Kim; Nexo, Ebba; Borghammer, Per
Introduction. Decreased salivary flow and xerostomia are frequent findings in Parkinson's disease (PD), possibly caused by alterations in the parasympathetic tonus. Here we explore salivary acetylcholinesterase (AChE) activity as a potential biomarker in PD. Methods. We measured salivary flow, AChE activity, and total protein concentration in 30 PD patients and 49 healthy controls. We also performed exploratory correlation analyses with disease duration, motor symptom severity, autonomic complaints, and other nonmotor symptoms. Results. PD patients displayed significantly decreased salivary flow rate, significantly increased salivary AChE activity, and total protein concentration. Importantly, the AChE activity/total protein ratio was significantly increased in PD patients, suggesting that increased AChE activity cannot be explained solely by upconcentration of saliva. The Unified PD Rating Scale (UPDRS) score displayed significant correlation with total salivary protein (P = 0.002) and near-significant correlation with salivary flow (P = 0.07). Color vision test scores were also significantly correlated with AChE activity (P = 0.04) and total protein levels (P = 0.002). Conclusion. Salivary AChE activity is increased in PD patients compared to healthy controls. Future studies are needed to elucidate whether this parameter reflects the extent of neuronal damage and parasympathetic denervation in the salivary glands of PD patients. PMID:25767737
Salivary acetylcholinesterase activity is increased in Parkinson's disease: a potential marker of parasympathetic dysfunction.
Fedorova, Tatyana; Knudsen, Cindy Soendersoe; Mouridsen, Kim; Nexo, Ebba; Borghammer, Per
Introduction. Decreased salivary flow and xerostomia are frequent findings in Parkinson's disease (PD), possibly caused by alterations in the parasympathetic tonus. Here we explore salivary acetylcholinesterase (AChE) activity as a potential biomarker in PD. Methods. We measured salivary flow, AChE activity, and total protein concentration in 30 PD patients and 49 healthy controls. We also performed exploratory correlation analyses with disease duration, motor symptom severity, autonomic complaints, and other nonmotor symptoms. Results. PD patients displayed significantly decreased salivary flow rate, significantly increased salivary AChE activity, and total protein concentration. Importantly, the AChE activity/total protein ratio was significantly increased in PD patients, suggesting that increased AChE activity cannot be explained solely by upconcentration of saliva. The Unified PD Rating Scale (UPDRS) score displayed significant correlation with total salivary protein (P = 0.002) and near-significant correlation with salivary flow (P = 0.07). Color vision test scores were also significantly correlated with AChE activity (P = 0.04) and total protein levels (P = 0.002). Conclusion. Salivary AChE activity is increased in PD patients compared to healthy controls. Future studies are needed to elucidate whether this parameter reflects the extent of neuronal damage and parasympathetic denervation in the salivary glands of PD patients. PMID:25767737
Aqueous extracts of Roselle (Hibiscus sabdariffa Linn.) varieties inhibit ?-amylase and ?-glucosidase activities in vitro.
Ademiluyi, Adedayo O; Oboh, Ganiyu
This study sought to investigate the inhibitory effect of aqueous extracts of two varieties (red and white) of Hibiscus sabdariffa (Roselle) calyces on carbohydrate hydrolyzing enzymes (?-amylase and ?-glucosidase), with the aim of providing the possible mechanism for their antidiabetes properties. Aqueous extracts were prepared (1:100 w/v) and the supernatant used for the analysis. The extracts caused inhibition of ?-amylase and ?-glucosidase activities in vitro.The IC(50) revealed that the red variety (25.2 ?g/mL) exhibited higher ?-glucosidase inhibitory activity than the white variety (47.4 ?g/mL), while the white variety (90.5 ?g/mL) exhibited higher ?-amylase inhibitory activity than the red variety (187.9 ?g/mL). However, the ?-glucosidase inhibitory activities of both calyces were higher than that of their ?-amylase. In addition, the red variety possessed higher antioxidant capacity as exemplified by the (•)OH scavenging abilities, Fe(2+) chelating ability, and inhibition of Fe(2+)-induced pancreatic lipid peroxidation in vitro. The enzyme inhibitory activities and antioxidant properties of the roselle extracts agreed with their phenolic content. Hence, inhibition of ?-amylase and ?-glucosidase, coupled with strong antioxidant properties could be the possible underlying mechanism for the antidiabetes properties of H. sabdariffa calyces; however, the red variety appeared to be more potent. PMID:23216107
?-AMYLASE4, a Noncatalytic Protein Required for Starch Breakdown, Acts Upstream of Three Active ?-Amylases in Arabidopsis Chloroplasts[W][OA
Fulton, Daniel C.; Stettler, Michaela; Mettler, Tabea; Vaughan, Cara K.; Li, Jing; Francisco, Perigio; Gil, Manuel; Reinhold, Heike; Eicke, Simona; Messerli, Gaëlle; Dorken, Gary; Halliday, Karen; Smith, Alison M.; Smith, Steven M.; Zeeman, Samuel C.
This work investigated the roles of ?-amylases in the breakdown of leaf starch. Of the nine ?-amylase (BAM)–like proteins encoded in the Arabidopsis thaliana genome, at least four (BAM1, -2, -3, and -4) are chloroplastic. When expressed as recombinant proteins in Escherichia coli, BAM1, BAM2, and BAM3 had measurable ?-amylase activity but BAM4 did not. BAM4 has multiple amino acid substitutions relative to characterized ?-amylases, including one of the two catalytic residues. Modeling predicts major differences between the glucan binding site of BAM4 and those of active ?-amylases. Thus, BAM4 probably lost its catalytic capacity during evolution. Total ?-amylase activity was reduced in leaves of bam1 and bam3 mutants but not in bam2 and bam4 mutants. The bam3 mutant had elevated starch levels and lower nighttime maltose levels than the wild type, whereas bam1 did not. However, the bam1 bam3 double mutant had a more severe phenotype than bam3, suggesting functional overlap between the two proteins. Surprisingly, bam4 mutants had elevated starch levels. Introduction of the bam4 mutation into the bam3 and bam1 bam3 backgrounds further elevated the starch levels in both cases. These data suggest that BAM4 facilitates or regulates starch breakdown and operates independently of BAM1 and BAM3. Together, our findings are consistent with the proposal that ?-amylase is a major enzyme of starch breakdown in leaves, but they reveal unexpected complexity in terms of the specialization of protein function. PMID:18390594
Detection of amylase activity from fruit and vegetables in an undergraduate classroom
Surasak Laloknam; Supaporn Sirisopana; Somkiat Phornphisutthimas
This research aimed to construct a hands-on activity for undergraduate students to understand how to detect and compare amylase activity from various sources by using a simple method. The amylolytic activity of extracts from 10 kinds of vegetables, Chinese white vegetable, tomato, cucumber, pumpkin, pea eggplant, carrot, cabbage, morning glory, Chinese broccoli, and yard long bean as well as 10
Comparative study of digestive enzymes in fish with different nutritional habits. Proteolytic and amylase activities
M. C Hidalgo; E Urea; A Sanz
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
Anticoagulant activities in salivary glands of tabanid flies.
Kazimírová, M; Sulanová, M; Trimnellt, A R; Kozánek, M; Vidlicka, L; Labuda, M; Nuttall, P A
Tabanid flies are telmophages (pool feeders), taking frequent and rapid bloodmeals from many different individual hosts. To investigate how they accomplish this intermittent feeding strategy, we examined the anticoagulant activities in salivary gland extracts (SGE) from 19 species representing six genera: Atylotus, Chrysops, Haematopota, Heptatoma, Hybomitra and Tabanus (Diptera: Tabanidae). Standard coagulation screen assays were used to determine thrombin time, prothrombin time and activated partial thromboplastin time. Chromogenic substrate assays were performed for thrombin and factor Xa activity. SGE of most species (except Chrysops spp.) considerably prolonged human plasma clotting time in a dose-dependent manner, and showed potent and specific antithrombin activity in the chromogenic substrate assay. Heptatoma pellucens displayed the strongest anticoagulant activity. Specific anti-factor Xa activity in tabanid SGE was not detected. Electrophoretic profiles of SGE proteins differed between genera and species. Overall, the results suggest that tabanids have evolved at least two antihaemostatic strategies. PMID:12243231
Modified alpha-amylase activity among insecticide-resistant and -susceptible strains of the maize weevil, Sitophilus zeamais.
Lopes, K V G; Silva, L B; Reis, A P; Oliveira, M G A; Guedes, R N C
Fitness cost is usually associated with insecticide resistance and may be mitigated by increased energy accumulation and mobilization. Preliminary evidence in the maize weevil (Coleoptera: Curculionidae) suggested possible involvement of amylases in such phenomenon. Therefore, alpha-amylases were purified from an insecticide-susceptible and two insecticide-resistant strains (one with fitness cost [resistant cost strain], and the other without it [resistant no-cost strain]). The main alpha-amylase of each strain was purified by glycogen precipitation and ion-exchange chromatography (>or=70-fold purification,
[Effect of tobacco smoking on amylase activity in patients with pancreatitis].
Milnerowicz, Halina; Sliwi?ska, Mariola; Jab?onowska, Monika; Milnerowicz, Stanis?aw
The pancreas is one of the first organs pathologically affected by the tobacco smoking. However, the mechanism of development of these changes is not eventually recognised. It has been demonstrated that nicotine influences exogenous function of pancreas. The aim of this study is to prove the influence of tobacco smoking on amylase activity in serum and urine of smoking and non-smoking patients with diagnosed acute (AP), chronic (CP) and chronic exaggerated pancreatitis (CEP). Serum and urine has been collected from 57 patients with AP, CP and CEP. The activity of enzyme has been determined using the colorimetric method with ethylidene-G7-PNP (4,6-ethylidene-p-nitrophenyl-alpha,D-malthoheptozyde). The nicotine metabolites has been assayed with the immunoenzymatic method (ELISA) using rabbit polyclonal antibodies against cotinine. The highest amylase activity in serum and urine has been observed in smoking patients with CEP. Much higher differences in amylase activity has been estimated in urine of patients with CP and AP (CP: in non-smoking patients more than three times lower activity than in smoking patients; AP: in non-smoking patients more than two times lower activity than in smoking patients). It has been revealed that the differences in amylase activity in both serum and urine in smoking patients in comparison with non-smoking patients with pancreatitis may prove a significant influence of tobacco smoking on exocrine function of pancreas. PMID:15794254
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...
Beta-Amylase activity and thermostability in wild and cultivated barleys with different Bmy1 intron III alleles
Technology Transfer Automated Retrieval System (TEKTRAN)
The third intron of barley (Hordeum vulgare L.) beta-amylase 1 (Bmy1) is extremely polymorphic. Specific insertion/deletions (indels) in the third intron have been correlated with beta-amylase activity and thermostability and may have potential as a selective marker for cultivar development. The B...
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...
Salivary alpha amylase diurnal pattern and stress response are associated with body mass index in low-income preschool-aged children.
Miller, Alison L; Sturza, Julie; Rosenblum, Katherine; Vazquez, Delia M; Kaciroti, Niko; Lumeng, Julie C
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
Changes in carbohydrate and nitrogenous components and amylase activities during germination of grain amaranth.
Balasubramanian, T; Sadasivam, S
Grain amaranth (Amaranthus hypochondriacus), Yercaud local variety, was soaked overnight and germinated for 192 h taking the soaked grains as the zero time (0 h) sample. The changes in the activities of alpha- and beta-amylases, starch, sugar, protein and lysine contents during germination are reported. Activity of alpha-amylase was high in the 0 h soaked grains, while beta-amylase activity was high in 72 h germinated grains. The joint action of the amylases resulted in a decrease of starch content from 0 to 192 h in germinated grains and an increase in total sugars during the initial period of germination. Protein nitrogen was found to decrease from 48 h to 192 h accompanied by an increase in free amino acid and non-protein nitrogen contents. Total lysine content was found to be increased by 31% in 24 h germinated grain amaranth. Protein fractionation of raw, soaked and 24 h germinated grain amaranth showed that the distribution of different types of proteins varied during germination of the grains. An increase of water soluble protein content was noticed in 24 h germinated grains. PMID:2483588
Quantitative gel-electrophoretic determination of serum amylase isoenzyme distributions.
Gillard, B K
I report a direct, sensitive, quantitative method for determining serum amylase isoenzyme activity with commercially available reagents. Day-to-day reproducibility (CV) was 3--4% for the isoenzymes in normal serum; within-run precision was 8, 3, and 2% for low, normal and high isoenzyme activities. Amylase isoenzymes, separated into the pancreatic and salivary types by electrophoresis in polyacrylamide gel, are then quantified by directly incubating the gels in soluble-starch solution, staining with iodine, and densitometry. The proportion of pancreatic isoenzyme (47 normal sera) was 43 +/- 8% (mean +/- SD). Isoenzyme activities as low as 2% of normal can be measured accurately in 10 micro L of serum. The reproducibility, precision, and sensitivity indicate that the method is applicable to differential diagnosis of hyperamylasemia or hypoamylasemia, and is suited for monitoring the subtle changes in serum amylase isoenzyme distribution that may accompany disease progression or therapy. PMID:498502
Partial characterization of cold active amylases and proteases of Streptomyces sp. from Antarctica
Cotârle?, Mihaela; Negoi??, Teodor Gh.; Bahrim, Gabriela E.; Stougaard, Peter
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
... 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 .
Salivary biomarkers of HPA axis and autonomic activity in adults with intellectual disability with and without stereotyped and self-injurious behavior disorders.
Symons, Frank J; Wolff, Jason J; Stone, Laura S; Lim, Tony K Y; Bodfish, James W
Salivary levels of biomarkers for the hypothalamic-pituitary-adrenal axis (HPA; cortisol) and sympatho-adreno-medullary system (SAM; ?-amylase) were measured in 51 adults (57% male) with neurodevelopmental disorders associated with intellectual disability (i.e., mental retardation) and chronic self-injurious behavior (SIB) and compared with matched controls without SIB. Cortisol levels differed significantly (p?0.01) between the SIB and control group (SIB?>?control). Within-group analyses showed significant differences (p?0.05) in levels of salivary ?-amylase between individuals with SIB and those with SIB meeting criteria for stereotyped movement disorder (SMD; SIB?+?SMD?>?SIB). Salivary ?-amylase was significantly correlated with frequency of stereotypy among the SIB group (r?=?0.36, p?0.05). These preliminary findings warrant further exploration into the role of the SAM system in the pathophysiology of SIB and related repetitive behaviors among individuals with neurodevelopmental disorders associated with intellectual disability. PMID:21484195
Grape seed and tea extracts and catechin 3-gallates are potent inhibitors of ?-amylase and ?-glucosidase activity.
Yilmazer-Musa, Meltem; Griffith, Anneke M; Michels, Alexander J; Schneider, Erik; Frei, Balz
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
Studies on activity, distribution, and zymogram of protease, ?-amylase, and lipase in the paddlefish Polyodon spathula.
Ji, H; Sun, H T; Xiong, D M
A series of biochemical determination and electrophoretic observations have been conducted to analyze the activities and characteristics of protease, ?-amylase, and lipase of paddlefish Polyodon spathula. The results obtained have been compared with those of bighead carp (Aristichthys nobilis) and hybrid sturgeon (Huso dauricus ? × Acipenser schrenki Brandt ?), in order to increase available knowledge of the physiological characteristics of this sturgeon species and to gain information with regard to its nutrition. Further, a comparative study of enzymatic activity, distribution, and characterization between commercial feed-reared paddlefish (CG) and natural live food-reared (NG) paddlefish was conducted. Results showed that higher proteolytic activity was observed in the pH range 2.5-3.0 and at a pH of 7.0 for paddlefish. Levels of acid protease activity of paddlefish were similar to that of hybrid sturgeon, and significantly higher than that of bighead carp. The inhibition assay of paddlefish showed that the rate of inhibition of tosyl-phenylalanine chloromethyl ketone was approximately 2.6-fold that of tosyl-lysine chloromethyl ketone. There was no significant difference observed for acid protease activity between PG and CG groups, whereas the activity of alkaline protease, ?-amylase, and lipase in the PG group were significantly lower than those in the CG group. The substrate sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis further showed that there were certain types of enzymes, especially ?-amylase, with similar molecular mass in the paddlefish and hybrid sturgeon. It can be inferred that acid digestion was main mechanism for protein hydrolysis in paddlefish, as reported for other fishes with a stomach. This indicates that the paddlefish requires higher alkaline protease, ?-amylase, and lipase activity to digest natural live food. PMID:21894570
Phenotypic Variation for Diastatic power, ß-Amylase Activity, and ß-Amylase Thermostability vs. Allelic Variation at the Bmy1 Locus in a Sample of North American Barley Germplasm
Technology Transfer Automated Retrieval System (TEKTRAN)
Malting quality data including diastatic power, ß-amylase activity, and ß-amylase thermostability, were collected on malts from three barley (Hordeum vulgare L.) breeding program trials containing two growth habits and 165 lines grown in multiple environments. We attempted to identify causal polymor...
Salivary biomarkers of HPA axis and autonomic activity in adults with intellectual disability with and without stereotyped and self-injurious behavior disorders
Frank J. Symons; Jason J. Wolff; Laura S. Stone; Tony K. Y. Lim; James W. Bodfish
Salivary levels of biomarkers for the hypothalamic–pituitary–adrenal axis (HPA; cortisol) and sympatho-adreno-medullary system\\u000a (SAM; ?-amylase) were measured in 51 adults (57% male) with neurodevelopmental disorders associated with intellectual disability\\u000a (i.e., mental retardation) and chronic self-injurious behavior (SIB) and compared with matched controls without SIB. Cortisol\\u000a levels differed significantly (p??control). Within-group analyses showed significant differences (p?0.05) in levels of salivary ?-amylase
Changes in salivary peroxidase and polymorphonuclear neutrophil leucocyte enzyme activities during the menstrual cycle.
Cockle, S M; Harkness, R A
An elevation in salivary peroxidase activity has been found about the time of ovulation in 14 menstrual cycles in a total of six women. This peak coincided with the ovulatory luteinizing hormone (LH) and oestrogen peak in the four cycles in which endocrine studies were performed. Rises in some polymorphonuclear neutrophil leucocyte enzymes were also seen around ovulation. The possible use of changes in salivary peroxidase as a method for detection of ovulation is discussed. PMID:708662
Bombesin Improves Adaptive Immunity of the Salivary Gland During Parenteral Nutrition
Pierre, Joseph F.; Heneghan, Aaron F.; Wang, Xinying; Roenneburg, Drew A.; Groblewski, Guy E.; Kudsk, Kenneth A.
Background The parotid and submandibular salivary glands are gut-associated lymphoid tissues (GALTs) that secrete immune compounds into the oral cavity. Parenteral nutrition (PN) without enteral stimulation decreases GALT function, including intestinal lymphocyte counts and secretory immunoglobulin A (sIgA) levels. Since the neuropeptide bombesin (BBS), a gastrin-releasing peptide analogue, stimulates intestinal function and restores GALT parameters, we hypothesized that PN + BBS would stimulate parotid and salivary gland IgA levels, T lymphocytes, and IgA plasma cell counts compared with PN alone. Methods Male (Institute of Cancer Research) ICR mice received intravenous catheters and were randomized to chow with saline, PN, or PN + BBS (15 µg/tid/mouse) for 5 days (8/group), 2 days after cannulation. Salivary glands were weighed and either frozen for IgA and amylase analysis or fixed for histological analysis of acinar cells, IgA+ plasma cells, and T lymphocytes. Small intestinal wash fluid was collected for IgA regression analysis with salivary glands. Results PN reduced organ weight, acinar cell size, and amylase activity compared with chow; BBS had no significant effects on these parameters. Compared with chow, PN significantly reduced salivary gland IgA levels, IgA+ plasma cells, and T lymphocytes. PN + BBS significantly elevated IgA and restored cellularity compared with PN. Salivary gland tissue homogenate IgA levels significantly correlated with intestinal fluid IgA levels. Conclusions Compared with chow, PN results in atrophy of the salivary glands characterized by reduced amylase, IgA, and immune cellularity. BBS has no effect on acinar cells or amylase activity compared with PN but maintains tissue IgA and plasma cells and T-lymphocyte numbers compared with chow. PMID:24121183
alpha-Amylase activity by the Beckman reaction system and suppression by pyruvate.
Hanson, N Q; Yasmineh, W G
We evaluated the new Beckman Enzymatic Amyalse-DS Method with maltotetraose as substrate. Our findings indicate that the method is approximately twice as sensitive as the old method involving soluble starch [Clin. Chem. 24, 762 (1978)]. The new method also shows a linear rate of reaction, in contrast to the curvilinear rate previously observed with the old method. The Km with maltotetraose is 0.77 g/L, or about twice that with soluble starch (0.42 g/L). The method correlates well with the Phadebas alpha-amylase method (r = 0.974 and 0.991 on 84 serum and 52 urine specimens, respectively). Of 43 specimens with high concentrations of pyruvate we examined for interference with alpha-amylase activity, only six showed interference when maltotetraose was the substrate. (With both Beckman methods the reaction of pyruvate with NADH produced lactate and NAD+ in the presence of lactate dehydrogenase as contaminant.) Pyruvate interference decreased with increases in (a) the alpha-amylase activity of the specimen, (b) the amount of NADH initially present in the maltotetraose reagent, and (c) the length of time the reconstituted maltotetroase reagent was allowed to stand before being used. PMID:222501
Using a Short Wavelength Infrared (SWIR) hyperspectral imaging system to predict alpha amylase activity in individual Canadian western wheat kernels
Juan Xing; Pham Van Hung; Stephen Symons; Muhammad Shahin; David Hatcher
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
?-Amylase sensor based on the degradation of oligosaccharide hydrogel films monitored with a quartz crystal sensor.
Gibbs, Martin John; Biela, Anna; Krause, Steffi
?-Amylase hydrolyses starch molecules to produce smaller oligosaccharides and sugars. Amylases are of great importance in biotechnology and find application in fermentation, detergents, food and the paper industry. The measurement of ?-amylase activity in serum and urine has been used in the diagnosis of acute pancreatitis. Salivary amylase has also been shown to be a stress indicator. Sensor coatings suitable for the detection of ?-amylase activity have been developed. Oligosaccharides such as glycogen and amylopectin were spin-coated onto gold coated quartz crystals with a base frequency of 10 MHz. The films were subsequently cross-linked with hexamethylene diisocyanate. Film degradation was monitored with a quartz crystal microbalance (QCM) and electrochemical impedance measurements. The films were shown to be stable in phosphate buffered saline (PBS). Addition of ?-amylase to the solution resulted in the rapid degradation of the films. The maximum rate of degradation was found to be strongly dependent on the amylase activity in the range typically found in serum when diagnosing pancreatitis (0.08-8 U/ml). Sensor responses in serum were found to be very similar to those obtained in buffer indicating the absence of non-specific binding. PMID:25266253
Apyrase and anti-platelet activities from the salivary glands of the bed bug Cimex lectularius
Jesus G. Valenzuela; Ouatfa M. Chuffe; JosÉM. C. Ribeiro
Apyrase activity was detected in the salivary gland of Cimex lectularius. Kinetic, HPLC chromatography, isoelectric focusing gel electrophoresis and thermal sensitivity assays indicated that the enzymatic activity is the result of a true apyrase and not the result of independent or combined different enzyme activities. The apyrase activity was dependent on calcium and not on magnesium, manganese or zinc, and
Study of alpha-amylase and urease inhibitory activities of Melilotus indicus (Linn.) All.
Ahmed, Dildar; Younas, Saba; Anwer-Mughal, Qaria Mumtaz
Melilotus indicus (Linn.) All. is a small herb distributed throughout Pakistan and has a number of ethnomedicinal uses. It is also consumed as a vegetable. In the present work, we are reporting the alpha-amylase and urease inhibitory activities of methanolic extract of M. indicus and its sub-fractions in different solvents. Both the methanolic extract and its fractions in chloroform, ethyl acetate, n-butanol and water showed remarkable inhibitory activities against alpha-amylase with the IC50 values being 1.29, 1.45, 1.07, 1.45 and 2.10 mg/mL respectively. The efficacy of the methanolic extract was comparable with that of acarbose (1.20 mg/mL), while the ethyl acetate fraction was more potent. The urease inhibitory activities of methanolic extract and chloroform, ethyl acetate, n-butanol and water fractions were more prominent with IC50 values being 0.95, 0.89, 1.53, 0.98 and 4.90 ?g/mL respectively. The activity of methanolic extract was slightly higher than that of thiourea (0.97 ?g/mL) which in turn was slightly higher than that of n-butanolic fraction. The chloroform fraction showed the highest anti-urease activity. All the plant samples showed enzyme inhibitory activity in a dose-dependent manner. Moreover, they were manifold more effective against urease than alpha-amylase. The combination of the plant extract with acarbose considerably increased the potency of the latter. The findings suggest that enzyme inhibitory activities of the vegetable M. indicus may have pharmacological significance against diabetes mellitus and gastrointestinal ulcers. PMID:24374453
Activation of bean (Phaseolus vulgaris) [alpha]-amylase inhibitor requires proteolytic processing of the proprotein
Pueyo, J.J.; Hunt, D.C.; Chrispeels, M.J. (Univ. of California, San Diego, La Jolla (United States))
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.
Evaluation of ten wild nigerian mushrooms for amylase and cellulase activities.
Jonathan, Segun Gbolagade; Adeoyo, Olusegun Richard
Amylases and cellulases are important enzymes that can be utilized for various biological activities. Ten different wild Nigerian mushrooms (Agaricus blazei, Agaricus sp., Corilopsis occidentalis, Coriolus versicolor, Termitomyces clypeatus, Termitomyces globulus, Pleurotus tuber-regium, Podoscypha bolleana, Pogonomyces hydnoides, and Nothopanus hygrophanus) were assayed for production of these secondary metabolites. The results revealed that most of the tested wild fungi demonstrated very good amylase and cellulase activities. With the incorporation of carboxymethyl-cellulose (a carbon source) into the culture medium, Agaricus blazei had the highest amylolytic activity of 0.60 unit/mL (at 25?, pH 6.8). This was followed in order by P. tuber-regium and Agaricus sp. with 0.42 and 0.39 unit/mL, respectively (p ? 0.05). Maltose and sucrose supplementation into the submerged liquid medium made N. hygrophanus and P. hydnoides to exhibit very low amylase activities of 0.09 and 0.11 unit/mL, respectively. Introducing peptone (an organic nitrogen source) into the basal medium enhanced the ability of C. versicolor to produce a cellulase value of 0.74 unit/mL. Other organic nitrogen sources that supported good cellulase activities were yeast extract and urea. Sodium nitrate (inorganic nitrogen source) generally inhibited cellulase production in all mushrooms. The best carbon source was carboxymethyl-cellulose, which promoted very high cellulase activity of 0.67 unit/mL in C. versicolor, which was followed in order by P. tuber-regium, T. chypeatus, and C. occidentalis (p ? 0.05). Sucrose was the poorest carbon compound, supporting the lowest values of 0.01, 0.01, and 0.14 unit/mL in P. hydnoides, A. blazei, and Agaricus sp., respectively. PMID:22783085
Aberrant Activation of the RANK Signaling Receptor Induces Murine Salivary Gland Tumors.
Szwarc, Maria M; Kommagani, Ramakrishna; Jacob, Allison P; Dougall, William C; Ittmann, Michael M; Lydon, John P
Unlike cancers of related exocrine tissues such as the mammary and prostate gland, diagnosis and treatment of aggressive salivary gland malignancies have not markedly advanced in decades. Effective clinical management of malignant salivary gland cancers is undercut by our limited knowledge concerning the key molecular signals that underpin the etiopathogenesis of this rare and heterogeneous head and neck cancer. Without knowledge of the critical signals that drive salivary gland tumorigenesis, tumor vulnerabilities cannot be exploited that allow for targeted molecular therapies. This knowledge insufficiency is further exacerbated by a paucity of preclinical mouse models (as compared to other cancer fields) with which to both study salivary gland pathobiology and test novel intervention strategies. Using a mouse transgenic approach, we demonstrate that deregulation of the Receptor Activator of NFkB Ligand (RANKL)/RANK signaling axis results in rapid tumor development in all three major salivary glands. In line with its established role in other exocrine gland cancers (i.e., breast cancer), the RANKL/RANK signaling axis elicits an aggressive salivary gland tumor phenotype both at the histologic and molecular level. Despite the ability of this cytokine signaling axis to drive advanced stage disease within a short latency period, early blockade of RANKL/RANK signaling markedly attenuates the development of malignant salivary gland neoplasms. Together, our findings have uncovered a tumorigenic role for RANKL/RANK in the salivary gland and suggest that targeting this pathway may represent a novel therapeutic intervention approach in the prevention and/or treatment of this understudied head and neck cancer. PMID:26061636
Aberrant Activation of the RANK Signaling Receptor Induces Murine Salivary Gland Tumors
Jacob, Allison P.; Dougall, William C.; Ittmann, Michael M.; Lydon, John P.
Unlike cancers of related exocrine tissues such as the mammary and prostate gland, diagnosis and treatment of aggressive salivary gland malignancies have not markedly advanced in decades. Effective clinical management of malignant salivary gland cancers is undercut by our limited knowledge concerning the key molecular signals that underpin the etiopathogenesis of this rare and heterogeneous head and neck cancer. Without knowledge of the critical signals that drive salivary gland tumorigenesis, tumor vulnerabilities cannot be exploited that allow for targeted molecular therapies. This knowledge insufficiency is further exacerbated by a paucity of preclinical mouse models (as compared to other cancer fields) with which to both study salivary gland pathobiology and test novel intervention strategies. Using a mouse transgenic approach, we demonstrate that deregulation of the Receptor Activator of NFkB Ligand (RANKL)/RANK signaling axis results in rapid tumor development in all three major salivary glands. In line with its established role in other exocrine gland cancers (i.e., breast cancer), the RANKL/RANK signaling axis elicits an aggressive salivary gland tumor phenotype both at the histologic and molecular level. Despite the ability of this cytokine signaling axis to drive advanced stage disease within a short latency period, early blockade of RANKL/RANK signaling markedly attenuates the development of malignant salivary gland neoplasms. Together, our findings have uncovered a tumorigenic role for RANKL/RANK in the salivary gland and suggest that targeting this pathway may represent a novel therapeutic intervention approach in the prevention and/or treatment of this understudied head and neck cancer. PMID:26061636
Determination of antioxidant capacity, ?-amylase and lipase inhibitory activity of Crotalaria juncea Linn in vitro inhibitory activity of Crotalaria Juncea Linn.
Dinakaran, Sathis Kumar; Banji, David; Avasarala, Harani; Banji, Otilia
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
Chloride Activated Halophilic ?-Amylase from Marinobacter sp. EMB8: Production Optimization and Nanoimmobilization for Efficient Starch Hydrolysis
Kumar, Sumit; Khare, S. K.
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
?-amylase from wheat (Triticum aestivum) seeds: its purification, biochemical attributes and active site studies.
Singh, Kritika; Kayastha, Arvind M
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
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 ...
Effect of fluoride preparations on the activity of human salivary cathepsin C.
Dabrowska, E; Letko, M; Roszkowska-Jakimiec, W; Letko, R; Jamio?kowski, J
Preparations containing organic and inorganic fluorine compounds are used for oral hygiene. Fluoride ions contained in these preparations display high bioactivity and can alter the environment of the mouth. The aim of the study was to determine the effect of preparations containing aminofluorides, commonly used in oral hygiene, on the activity of salivary cathepsin C (EC 126.96.36.199). The research material included mixed saliva, collected at rest before and after the application of the following preparations: Elmex gelee, Elmex red fluid, Elmex green fluid, Fluormex rinse. The salivary pH, concentration of fluoride ions and activity of cathepsin C were determined. Fluoride preparations inhibit the activity of cathepsin C and cause changes in human salivary pH. Saliva can serve as a diagnostic material in the examination of the environmental exposure to fluorides. PMID:16119654
[Effect of pectin substances on activity of human pancreatic alpha-amylase in vitro].
Chelpanova, T I; Vitiazev, F V; Mikhaleva, N Ia; Efimtseva, É A
Pectin substances were extracted from food plants: sweet pepper Capsicum annuum L., carrot sowing Daucus sativus L., bulb onion Allium cepa L., white cabbage Brassica oleracea L. by two methods with acid solutions similar to gastric environment. The pectins that were extracted were characterized by Monosaccharide composition and quantitative contents of uronic acids, neutral monosaccharides, methoxy groups, protein. The inhibitory effect of all extracted pectin-protein complexes on activity of pharmaceutical drugs of human pancreatic alpha-amylase was detected. It was found that the inhibitory effect of isolated pectin substances was dependent upon the species of plant source, the manner of pectin substance extraction, the chemical composition and acting concentrations. The ability of pectin substances to suppress enzyme activity was found in a range of pectin concentrations from 0.5 up to 1.5 %. It was revealed that extracted pectin substances from bulb onion and white cabbage by acid solution with pepsin had a 2.4-3.4 times greater inhibiting effect on the human pancreatic alpha-amylase activity in comparison with pectin substances extracted by solution without pepsin from the same plant sources in high concentrations. PMID:23013011
Chemical modification of liquefying alpha-amylase: role of tyrosine residues at its active center.
Kochhar, S; Dua, R D
Liquefying alpha-amylase from Bacillus amyloliquefaciens was inactivated by treatment with tetranitromethane and N-acetylimidazole. The loss of activity occurred with modification of five tyrosine residues. Preincubation of the enzyme with either the substrate or the competitive inhibitor at saturating levels provided complete protection against inactivation. However, the presence of substrate/inhibitor in the reaction mixture protected only two of the five modifiable tyrosine residues, suggesting the involvement of only two tyrosine residues at the active center. This was confirmed when hydroxylamine treatment of the acetylated enzyme fully restored the enzymatic activity. Both nitration and acetylation increased the apparent Km of the enzyme for soluble starch, which indicated that the tyrosine residues are involved in substrate binding. Reduction of nitrotyrosine residues to aminotyrosine residues failed to restore the enzymatic activity. So, the loss of activity on modification of tyrosine residues was ascribed to conformational perturbances and not simply to the changes in the ionic character of tyrosine residues. PMID:3875315
Enhanced antifungal and insect ?-amylase inhibitory activities of Alpha-TvD1, a peptide variant of Tephrosia villosa defensin (TvD1) generated through in vitro mutagenesis.
Vijayan, S; Imani, J; Tanneeru, K; Guruprasad, L; Kogel, K H; Kirti, P B
TvD1 is a small, cationic, and highly stable defensin from the weedy legume, Tephrosia villosa with demonstrated in vitro antifungal activity. We show here peptide modifications in TvD1 that lead to enhanced antifungal activities. Three peptide variants, S32R, D37R, and Alpha-TvD1 (-G-M-T-R-T-) with variations in and around the ?2-?3 loop region that imposes the two ?-strands, ?2 and ?3 were generated through in vitro mutagenesis. Alpha-TvD1 exhibited enhanced antifungal activity against the fungal pathogens, Fusarium culmorum and Fusarium oxysporum with respective IC(50) values of 2.5 ?M and 3.0 ?M, when compared to S32R (<5.0 ?M and >5.0 ?M), D37R (5.5 ?M and 4.5 ?M), and the wild type TvD1 (6.5 ?M). Because of the enhanced antifungal activity, this variant peptide was characterized further. Growth of F. culmorum in the presence of Alpha-TvD1 showed deformities in hyphal walls and nuclear damage. With respect to the plant pathogenic bacterium, Pseudomonas syringae pv. tomato strain DC3000, both Alpha-TvD1 and the wild type TvD1 showed comparable antibacterial activity. Both wild type TvD1 and Alpha-TvD1 displayed inhibitory activity against the ?-amylase of the mealworm beetle, Tenebrio molitor (TMA) with the latter showing enhanced activity. The human salivary as well as barley ?-amylase activities were not inhibited even at concentrations of up to 50 ?M, which has been predicted to be due to differences in the pocket size and the size of the interacting loops. Present study shows that the variant Alpha-TvD1 exhibits enhanced antifungal as well as insect ?-amylase inhibitory activity. PMID:22244814
Concurrent attenuated reactivity of alpha-amylase and cortisol is related to disruptive behavior in male adolescents.
de Vries-Bouw, Marjan; Jansen, Lucres; Vermeiren, Robert; Doreleijers, Theo; van de Ven, Peter; Popma, Arne
Attenuated reactivity of salivary alpha-amylase has been proposed as a specific sympathetic marker of disruptive behavior in juveniles and may have additional value to studying other autonomic parameters and hypothalamic-pituitary-adrenal axis activity. Investigating the interrelationships between neurobiological parameters in relation to juvenile disruptive behavior may enhance insight into the complex mechanisms at play. We investigated salivary alpha-amylase, cortisol, heart rate (HR), and heart rate variability (HRV) in response to a standardized public speaking task, and examined interactions between these parameters in relation to disruptive behavior. Participants were 48 delinquent male adolescents (mean age 18.4 years, SD 0.9), with and without a disruptive behavior disorder (resp. DP+, DP-) and 16 matched normal controls (NC). A structured psychiatric interview as well as the Youth Self Report and Child Behavior Checklist were administered to assess disruptive behavior. Alpha-amylase and cortisol reactivity, but not HR or HRV, showed significant inverse associations with dimensional measures of disruptive behavior. Moreover, both cortisol and alpha-amylase reactivity were significantly lower in the DP+ group as compared to the NC group. The mentioned relationships remained present when nicotine use was entered as a covariate. Combining alpha-amylase and cortisol in one model explained a larger part of the variance of disruptive behavior than either single parameter. There were no interactions between alpha-amylase and cortisol or HRV in relation to disruptive behavior. Attenuated alpha-amylase responsivity to stress is a correlate of disruptive behavior in late-adolescent males. Although nicotine use explains a considerable part of the variance of disruptive behavior, both alpha-amylase and cortisol are related to disruptive behavior, over and above the effect of nicotine use. Combining alpha-amylase and cortisol improved insight into neurobiological mechanisms involved with disruptive behavior; concurrent low reactivity of both parameters was related to higher levels of disruptive behavior. PMID:22587939
Salivary enzymes in peptic ulcer disease
Motamedi, Mojdeh; Mansour-Ghanaei, Fariborz; Sariri, Reyhaneh; Vesal, Mahmoud
Aim Peptic ulcer, the common disease of the upper gastro-intestinal tract, occurs in about 5–10% of the world's population. Therefore, diagnosis of trace disease progression with a noninvasive method is of prime importance in the field of healthcare research. The aim of this study was to evaluate the validity of salivary enzymes as noninvasive biomarkers for peptic ulcer. Materials and methods In practice, 34 peptic ulcer patients and 30 healthy subjects donated their un-stimulated saliva samples after 8 h of fasting. The activity of some selected enzymes was measured using appropriate enzymatic assay methods. Results The results indicated an overall alternation in enzymatic activity of saliva in patients suffering from peptic ulcer. Biological activity of a-amylase, peroxidase and lactate dehydrogenase, showed significantly higher values in almost all patients as compared to control subjects. Conclusions Based on the results of salivary enzyme activity, it was concluded that besides the influence of their peptic ulcer on enzyme activity of saliva, the considerably higher activity of a-amylase could also be related to the major role of the enzyme on physiological oxidative stress. PMID:25737890
The in vitro evaluation of antioxidative activity, ?-glucosidase and ?-amylase enzyme inhibitory of natural phenolic extracts.
Djeridane, Amar; Hamdi, Aicha; Bensania, Wafa; Cheifa, Khadidja; Lakhdari, Imane; Yousfi, Mohamed
Phenolic extracts from the medicinal parts of six traditional Algerian herbs were tested in screening experiments for the antioxidant, ?-amylase and ?-glycosidase inhibiting activities. UV-analysis of the extracts from the plants indicated that the total phenols content was ranged between 0.48 and 3.46mg equivalent of gallic acid per gram of dry matter, whereas the flavonoids content expressed as rutin equivalent per gram of dry matter was ranged between 0.18 and 2.23mg/g. The study of antioxidant activity by scavenging the hydroxyl radical (OH), the nitroxide radical (NO) and the stable radical cation (ABTS(+)) showed a high antioxidant power. Also, these extracts illustrated a significant reductive power of the Fe(3+)-TPTZ complex. Similarly, we have found that the phenolic extracts exhibit an imperative antioxidant status compared to synthetic antioxidants. The study of the extract effects shows that Anabasis articulata, Agatophora alopecuroide and Heliantheum kahiricum extracts have a powerful inhibiting capacity of the ?-amylase and ?-glycosidase with a Ki values less than 10?M. Our study, for the first time, revealed the anti-diabetic potential of the six plants and the results of this study could be helpful to develop medicinal preparations or nutraceuticals and functional foods for diabetes. PMID:25470628
Salivary aldehyde dehydrogenase - temporal and population variability, correlations with drinking and smoking habits and activity towards aldehydes contained in food.
Giebu?towicz, Joanna; Dziadek, Marta; Wroczy?ski, Piotr; Wo?nicka, Katarzyna; Wojno, Barbara; Pietrzak, Monika; Wierzchowski, Jacek
Fluorimetric method based on oxidation of the fluorogenic 6-methoxy-2-naphthaldehyde was applied to evaluate temporal and population variability of the specific activity of salivary aldehyde dehydrogenase (ALDH) and the degree of its inactivation in healthy human population. Analyzed was also its dependence on drinking and smoking habits, coffee consumption, and its sensitivity to N-acetylcysteine. Both the specific activity of salivary ALDH and the degree of its inactivation were highly variable during the day, with the highest activities recorded in the morning hours. The activities were also highly variable both intra- and interpersonally, and negatively correlated with age, and this correlation was stronger for the subgroup of volunteers declaring abstinence from alcohol and tobacco. Moderately positive correlations of salivary ALDH specific activity with alcohol consumption and tobacco smoking were also recorded (r(s) ~0.27; p=0.004 and r(s) =0.30; p=0.001, respectively). Moderate coffee consumption correlated positively with the inactivation of salivary ALDH, particularly in the subgroup of non-drinking and non-smoking volunteers. It was found that mechanical stimulation of the saliva flow increases the specific activity of salivary ALDH. The specific activity of the salivary ALDH was strongly and positively correlated with that of superoxide dismutase, and somewhat less with salivary peroxidase. The antioxidant-containing drug N-acetylcysteine increased activity of salivary ALDH presumably by preventing its inactivation in the oral cavity. Some food-related aldehydes, mainly cinnamic aldehyde and anisaldehyde, were excellent substrates of the salivary ALDH3A1 enzyme, while alkenals, particularly those with short chain, were characterized by lower affinity towards this enzyme but high catalytic constants. The protective role of salivary ALDH against aldehydes in food and those found in the cigarette smoke is discussed, as well as its participation in diminishing the effects of alcohol- and smoking-related oxidative stress. PMID:20931090
Human ?-amylase present in lower-genital-tract mucosal fluid processes glycogen to support vaginal colonization by Lactobacillus.
Spear, Gregory T; French, Audrey L; Gilbert, Douglas; Zariffard, M Reza; Mirmonsef, Paria; Sullivan, Thomas H; Spear, William W; Landay, Alan; Micci, Sandra; Lee, Byung-Hoo; Hamaker, Bruce R
Lactobacillus colonization of the lower female genital tract provides protection from the acquisition of sexually transmitted diseases, including human immunodeficiency virus, and from adverse pregnancy outcomes. While glycogen in vaginal epithelium is thought to support Lactobacillus colonization in vivo, many Lactobacillus isolates cannot utilize glycogen in vitro. This study investigated how glycogen could be utilized by vaginal lactobacilli in the genital tract. Several Lactobacillus isolates were confirmed to not grow in glycogen, but did grow in glycogen-breakdown products, including maltose, maltotriose, maltopentaose, maltodextrins, and glycogen treated with salivary ?-amylase. A temperature-dependent glycogen-degrading activity was detected in genital fluids that correlated with levels of ?-amylase. Treatment of glycogen with genital fluids resulted in production of maltose, maltotriose, and maltotetraose, the major products of ?-amylase digestion. These studies show that human ?-amylase is present in the female lower genital tract and elucidates how epithelial glycogen can support Lactobacillus colonization in the genital tract. PMID:24737800
Comparison of Salivary Beta Glucuronidase Activity in Chronic Periodontitis Patients with and without Diabetes Mellitus
ND, Jayakumar; Varghese, Sheeja
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
Draft Genome Sequences of 18 Oral Streptococcus Strains That Encode Amylase-Binding Proteins
Sabharwal, Amarpreet; Liao, Yu-Chieh; Lin, Hsin-Hung; Haase, Elaine M.
A number of commensal oral streptococcal species produce a heterogeneous group of proteins that mediate binding of salivary ?-amylase. This interaction likely influences streptococcal colonization of the oral cavity. Here, we present draft genome sequences of several strains of oral streptococcal species that bind human salivary amylase. PMID:25999552
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 ...
Interindividual variation, correlations, and sex-related differences in the salivary biochemistry of young healthy adults.
Prodan, Andrei; Brand, Henk S; Ligtenberg, Antoon J M; Imangaliyev, Sultan; Tsivtsivadze, Evgeni; van der Weijden, Fridus; Crielaard, Wim; Keijser, Bart J F; Veerman, Enno C I
A cross-sectional observational study was conducted to evaluate interindividual biochemical variation in unstimulated whole saliva in a population of 268 systemically healthy young students, 18-30 yr of age, with no apparent caries lesions or periodontal disease. Salivary flow rate, protein content, pH, buffering capacity, mucins MUC5B and MUC7, albumin, secretory IgA, cystatin S, lactoferrin, chitinase, amylase, lysozyme, and proteases were measured using ELISAs and enzymatic activity assays. Significant differences were found between male and female subjects. Salivary pH, buffering capacity, protein content, MUC5B, secretory IgA, and chitinase activity were all lower in female subjects compared with male subjects, whereas MUC7 and lysozyme activity were higher in female subjects. There was no significant difference between sexes in salivary flow rate, albumin, cystatin S, amylase, and protease activity. Principal component analysis (PCA) and spectral clustering (SC) were used to assess intervariable relationships within the data set and to identify subgroups. Spectral clustering identified two clusters of participants, which were subsequently described. This study provides a comprehensive overview of the distribution and inter-relations of a set of important salivary biochemical variables in a systemically healthy young adult population, free of apparent caries lesions and periodontal disease. It highlights significant gender differences in salivary biochemistry. PMID:25809904
Antioxidant and ?-amylase inhibitory activities of extract and isolates from Zygogynum pancheri subsp. arrhantum.
Keskes, Henda; Litaudon, Marc; Cherif, Atef; Belhadj, Sahla; Hamdi, Besma; El Feki, Abdelfattah; Dumontet, Vincent; Ben Salah, Abdelhamid; Damak, Mohamed; Allouche, Noureddine
One new sesquiterpenoid (5R(*),8R(*),9R(*),10R(*))-cinnamolide (8), and seven known compounds, 5-hydroxy-7-methoxyflavonone (1), 8-hydroxy-3-(4'-hydroxyphenyl)-6,7-(2?,2?-dimethylchromene)-tetralone (2), 8-hydroxy-3-(3',4'-dihydroxyphenyl)-6,7-(2?,2?-dimethylchromene)-tetralone (3), 1?-E-O-p-methoxycinnamoyl-bemadienolide (4), 1?-O-(E-cinnamoyl)-6?-hydroxy-9-epi-polygodial (5), 1?-O-(E-cinnamoyl)-6?-hydroxypolygodial (6), and 1?-O-E-cinnamoylpolygodial (7) were isolated from the ethyl acetate extract of barks of Zygogynum pancheri subsp. arrhantum (Winteraceae). The structures of these molecules were assigned predominantly based on spectral data. The structure of compound 8 was confirmed by X-ray crystallographic analysis. Compounds 2 and 3 exhibited significant antioxidant activity, whereas compounds 1 and 4-7 showed significant ?-amylase inhibitory activity. PMID:25034255
Prednisone treatment alters the serum amylase and lipase activities in normal dogs without causing pancreatitis.
Fittschen, C; Bellamy, J E
In order to test the hypothesis that treatment with glucocorticoids causes pancreatitis in dogs, 18 mongrel dogs were divided into three groups of six individuals, each group receiving prednisone at different doses orally or intramuscularly for two weeks. Two groups consisting of six dogs each served as controls. Treatment for two weeks with oral prednisone at 1.2 mg/kg body weight or at 4 mg/kg body weight daily decreased the serum amylase activities, but increased the serum lipase activities. Postmortem examinations revealed microscopic evidence of mild pancreatitis in only one dog given prednisone, that clinically appeared normal. It was concluded that daily doses of 4 mg prednisone/kg body weight or less given orally or intramuscularly for two weeks do not cause pancreatitis in dogs. PMID:6202383
Pharmacological activation of the EDA/EDAR signaling pathway restores salivary gland function following radiation-induced damage.
Hill, Grace; Headon, Denis; Harris, Zoey I; Huttner, Kenneth; Limesand, Kirsten H
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
Determination of antioxidant capacity and a-amylase inhibitory activity of the essential oils from citronella grass and lemongrass
Technology Transfer Automated Retrieval System (TEKTRAN)
The objective of the present study was to determine the antioxidant capacity of and in vitro a-amylase inhibitory activity of the essential oils extracted from citronella grass and lemongrass. The chemical composition of the extracted essential oils was determined by GC-MS. The antioxidant capacity ...
Genomic architecture of alpha-amylase activity in mature rye grain relative to that of preharvest sprouting.
Masoj?, Piotr; Wi?niewska, Magdalena; ?a?, Anna; Milczarski, Pawe?; Berdzik, Marcin; P?dziwiatr, Daniel; Pol-Szyszko, Magdalena; Ga??za, Monika; Owsianicki, Rados?aw
Bi-directional selective genotyping (BSG) carried out on two opposite groups of F(9)(541?×?Ot1-3) recombinant inbred lines (RILs) with extremely low and extremely high alpha-amylase activities in mature (dry) grain of rye, followed by molecular mapping, revealed a complex system of selection-responsive loci. Three classes of loci controlling alpha-amylase activity were discerned, including four major AAD loci on chromosomes 3R (three loci) and 6RL (one locus) responding to both directions of the disruptive selection, 20 AAR loci on chromosomes 2RL (three loci), 3R (three loci), 4RS (two loci), 5RL (three loci), 6R (two loci) and 7R (seven loci) responding to selection for low alpha-amylase activity and 17 AAE loci on chromosomes 1RL (seven loci), 2RS (two loci), 3R (two loci), 5R (two loci) and 6RL (four loci) affected by selection for high alpha-amylase activity. The majority of the discerned AA loci also showed responsiveness to selection for preharvest sprouting (PHS). Two AAD loci on chromosome arm 3RL coincided with PHSD loci. The AAD locus on chromosome arm 3RS was independent from PHS, whereas that on chromosome 6RL belonged to the PHSR class. AAR-PHSR loci were found on chromosomes 4RS (one locus) and 5R (two loci) and AAE-PHSE loci were identified on chromosomes 1RL (one locus) and 5RL (one locus). Some PHSD loci represented the AAE (chromosomes 1RL, 3RS and 3RL) or AAR classes (chromosome 5RL). AAR and AAE loci not related to PHS were found on chromosomes 1RL, 2R, 3RS, 4R, 6RL and 7RL. On the other hand, several PHS loci (1RL, 3RS, 5RL, 6RS and 7RS) had no effect on alpha-amylase activity. Allele originating from the parental line 541 mapped in six AA loci on chromosomes 2R (two loci), 5R (three loci) and 7R (one locus) exerted opposite effects on PHS and alpha-amylase activity. Differences between the AA and PHS systems of loci may explain the weak correlation between these two traits observed among recombinant inbred lines. Strategies for the breeding of sprouting-resistant varieties with low alpha-amylase and high PHS resistance are discussed. PMID:21225388
Diet and the evolution of human amylase gene copy number variation
Sorenson, Michael
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
Aleurones from a Barley with Low [alpha]-Amylase Activity Become Highly Responsive to Gibberellin When Detached from the Starchy Endosperm.
Skadsen, R. W.
The physiological and molecular bases for contrasting [alpha]-amylase phenotypes were examined in germinating seeds of two barley (Hordeum vulgare L.) cultivars, Morex and Steptoe. Morex is a high-quality malting barley that develops high [alpha]-amylase activity soon after germination. Steptoe is a feed barley that develops only low [alpha]-amylase activity levels during this period. The expression of all high- and low-isoelectric point (pl) [alpha]-amylase isozymes is reduced in Steptoe. The amount of [alpha]-amylase mRNA per gram of seedling tissue is correspondingly lower in Steptoe. Southern blot analysis revealed that the cultivars have the same copy number and organization for most high- and low-pl genes. Steptoe seedlings or embryoless half-seeds produce little [alpha]-amylase in response to exogenous applications of gibberellic acid (GA3) compared with Morex. However, when isolated aleurones of both cultivars are treated with GA3, they produce similar amounts of high- and low-pl [alpha]-amylase RNAs. This suggests that a factor in the starchy endosperm is responsible for lowered [alpha]-amylase response in Steptoe. The factor is probably not abscisic acid (ABA), since the two cultivars have similar concentrations of ABA during germination. PMID:12231810
Factors affecting antimicrobial activity of MUC7 12-mer, a human salivary mucin-derived peptide
Guo-Xian Wei; Alexander N Campagna; Libuse A Bobek
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
Neohesperidin dihydrochalcone: presentation of a small molecule activator of mammalian alpha-amylase as an allosteric effector.
Kashani-Amin, Elaheh; Larijani, Bagher; Ebrahim-Habibi, Azadeh
Flavonoids and their precursor trans-chalcone have been reported as inhibitors of mammalian alpha-amylase. With regard to this background, neohesperidin dihydrochalcone (NHDC) effect was investigated toward porcine pancreatic alpha-amylase (PPA), and found to be an activator of the enzyme. The maximal activation (up to threefold) was found to occur at 4.8mM of NHDC, which could be considered to have a high activation profile, with regard to the alpha and beta parameters (alpha<1
Perlecan domain IV peptide stimulates salivary gland cell assembly in vitro.
Pradhan, Swati; Zhang, Chu; Jia, Xinqiao; Carson, Daniel D; Witt, Robert; Farach-Carson, Mary C
Treatment of xerostomia would benefit from development of a functional implantable artificial salivary gland. Salivary gland tissue from surgical patients was assessed by histology and immunohistochemistry to establish the phenotype of normal salivary gland cells including the native basement membranes. Ductal and acinar cells were identified in tissue and cultured cells from dispersed tissue. High levels of laminin and perlecan/HSPG2 (heparan sulfate proteoglycan 2) 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 or a bioactive peptide derived from domain IV of perlecan. 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 found in tissue, were expressed in cultured acinar cells. Cells cultured on Matrigel or domain IV of perlecan peptide organized stress fibers and activated focal adhesion kinase. We report a novel technique to isolate acinar cells from human salivary gland and identify a human peptide sequence in perlecan that triggers differentiation of salivary gland cells into self-assembling acini-like structures that express essential biomarkers and which secrete alpha-amylase. PMID:19382872
Preferential salivary-type hypoamylasemia in obese children.
Mizuuchi, H; Taketa, K
Serum levels of total amylase, pancreatic type (P-type) isoamylase, and salivary type (S-type) isoamylase were measured in obese children (153 subjects; mean age, 10.1 years old; 86 boys and 67 girls) before and after weight reduction therapy. Serum amylase activities were determined using p-nitrophenylmaltoheptaoside as a substrate, with or without an antibody added to inhibit the S-type isoamylase. Serum levels of total amylase, P-type isoamylase and S-type isoamylase activities were significantly decreased in obese children with an obesity index more than 50%. S-type and P-type isoamylases showed negative correlation with the obesity index, the correlation coefficient being slightly larger in S-type than in P-type isoamylase. Analysis of the serum amylase activities in obese children who underwent weight reduction treatments showed a negative correlation only between the differences in S-type isoamylase activity and the differences in the obesity index. It may be concluded that the S-type isoamylase activity in serum of obese children is decreased and that it can be increased by reducing their body weight. PMID:10410788
A Novel Inhibitor of Factor X Activation from the Salivary Glands of the Bed Bug Cimex lectularius
Jesus G. Valenzuela; Jorge A. Guimaraes; José M. C. Ribeiro
Valenzuela, J. G., Guimaraes, J. A., and Ribeiro, J. M. C. 1996. A novel inhibitor of factor X activation from the salivary glands of the bed bugCimex lectularius. Experimental Parasitology83,184–190.Cimex lectulariussalivary gland homogenate delayed the recalcification time of human citrated plasma. Separation of the salivary gland homogenate by molecular sieving HPLC chromatography resulted in a single major peak of anticlotting
Hypoglycemic activity of Pyrus biossieriana Buhse leaf extract and arbutin: Inhibitory effects on alpha amylase and alpha glucosidase
Yousefi, Fatemeh; Mahjoub, Soleiman; Pouramir, Mahdi; Khadir, Fatemeh
Background: The mechanism of hypoglycemic and hypolipidemic activities of Pyrus biossieriana Buhse leaf extract (PbBLE) and its phytochemical component arbutin, have not been well determined. The present study was performed to understand the hypoglycemic activity mechanisms of pbBLE and arbutin more clearly. Methods: In vitro enzymatic carbohydrate digestion with PbBLE and arbutin was assessed using ?-amylase and ?-glucosidase powders. The enzyme solutions were premixed with PbBLE and arbutin at different concentrations (0.1, 1, 10 and 100 mg/ml). Substrate solutions and colorimetric reagents were added to the reaction. The release of glucose was determined by spectrophotometric method. Acarbose was used as the positive control. Results: The extract (10, 100 mg/ ml) completely inhibit ?- amylase and ?- glucosidase activities. The extract produced higher reduction of ?-amylase and ?-glucosidase activity than arbutin. Inhibition at various concentrations (0.1, 1, 10, 100 mg/ml) were significantly different (p<0.05). Conclusion: Our results exhibited that both the extract and arbutin were able to suppress the enzymes strongly. PMID:24294470
Detergent-compatible bacterial amylases.
Niyonzima, Francois N; More, Sunil S
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
Enzymatic activities in different strains isolated from healthy and brittle leaf disease affected date palm leaves: study of amylase production conditions.
Mouna, Jrad; Imen, Fendri; Choba Ines, Ben; Nourredine, Drira; Adel, Kadri; Néji, Gharsallah
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
Adaptive significance of amylase polymorphism in Drosophila.
Paris-Sud XI, Université de
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
Impaired Histatin-5 Levels and Salivary Antimicrobial Activity against C. albicans in HIV Infected Individuals
Khan, Shariq A; Fidel, Paul L; Thunayyan, Awdah Al; Varlotta, Sharon; Meiller, Timothy F; Jabra-Rizk, Mary Ann
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
Identification and analysis of a gene (abpA) encoding a major amylase-binding protein in Streptococcus gordonii.
Rogers, J D; Haase, E M; Brown, A E; Douglas, C W; Gwynn, J P; Scannapieco, F A
Oral streptococci such as Streptococcus gordonii bind the abundant salivary enzyme alpha-amylase. This interaction may be important in dental plaque formation and metabolism, thus contributing to the initiation and progression of dental caries and periodontal disease, the two most common plaque-mediated diseases. The conjugative transposon Tn916 was used to insertionally inactivate gene(s) essential to the expression of amylase-binding components of S. gordonii Challis, and a mutant deficient in amylase-binding (Challis Tn1) was identified. While wild-type strains of S. gordonii released both 20 kDa and 82 kDa amylase-binding proteins into culture supernatants, Challis Tn1 expressed the 82 kDa but not the 20 kDa protein. The 20 kDa amylase-binding protein was isolated from culture supernatants of S. gordonii Challis by hydroxyapatite chromatography. A partially purified, functionally active 20 kDa protein was sequenced from blots, and the N-terminal sequence obtained was found to be DEP(A)TDAAT(R)NND. A novel strategy, based on the single-specific-primer polymerase chain reaction technique, enabled the gene inactivated by Tn916 to be cloned. Analysis of the resultant nucleotide sequence revealed an open reading frame of 585 bp, designated amylase-binding protein A (abpA), encoding a protein of 20 kDa (AbpA), immediately downstream from the insertion site of Tn916. This protein possessed a potential signal peptide followed by a region having identity with the N-terminal sequence of the 20 kDa amylase-binding protein. These results demonstrate the role of the 20 kDa protein in the binding of amylase to S. gordonii. Knowledge of the nature of amylase-binding proteins may provide a better understanding of the role of these proteins in the colonization of S. gordonii in the oral cavity. PMID:9611797
Ca2(+)-dependent amylase secretion from pancreatic acinar cells occurs without activation of phospholipase C linked G-proteins.
Padfield, P J; Ding, T G; Jamieson, J D
We have examined the influence of guanine nucleotides on Ca2(+)-dependent amylase secretion from SLO permeabilized rat pancreatic acini. GTP gamma S (100 microM) stimulated Ca2+ dependent amylase release, decreasing the EC50 for Ca2+ from 1.4 to 0.8 microM. By contrast, GDP (1mM) and dGDP (1mM) inhibited the maximal Ca2(+)-dependent secretory response. Measurement of IP3 liberation showed that Ca2+ stimulation did not increase the activity of phospholipase C (PLC) postulated to be linked to a G-protein termed Gp; GDP and dGDP must therefore be exerting their inhibitory action via a GTP-binding protein distinct from the PLC-linked Gp. PMID:1899567
Wheat amylase trypsin inhibitors drive intestinal inflammation via activation of toll-like receptor 4
Junker, Yvonne; Zeissig, Sebastian; Kim, Seong-Jun; Barisani, Donatella; Wieser, Herbert; Leffler, Daniel A.; Zevallos, Victor; Libermann, Towia A.; Dillon, Simon; Freitag, Tobias L.; Kelly, Ciaran P.
Ingestion of wheat, barley, or rye triggers small intestinal inflammation in patients with celiac disease. Specifically, the storage proteins of these cereals (gluten) elicit an adaptive Th1-mediated immune response in individuals carrying HLA-DQ2 or HLA-DQ8 as major genetic predisposition. This well-defined role of adaptive immunity contrasts with an ill-defined component of innate immunity in celiac disease. We identify the ?-amylase/trypsin inhibitors (ATIs) CM3 and 0.19, pest resistance molecules in wheat, as strong activators of innate immune responses in monocytes, macrophages, and dendritic cells. ATIs engage the TLR4–MD2–CD14 complex and lead to up-regulation of maturation markers and elicit release of proinflammatory cytokines in cells from celiac and nonceliac patients and in celiac patients' biopsies. Mice deficient in TLR4 or TLR4 signaling are protected from intestinal and systemic immune responses upon oral challenge with ATIs. These findings define cereal ATIs as novel contributors to celiac disease. Moreover, ATIs may fuel inflammation and immune reactions in other intestinal and nonintestinal immune disorders. PMID:23209313
NSDL National Science Digital Library
Keith D. Stanley (Beloit College; Biology)
Information about sequence, structure, and active sites for many proteins is accessible online. This real world application of bioinformatics introduces us to amylases and asks us to explore functionally similar enzymes from markedly divergent organisms. Here, we search for microbial enzymes useful in the starch processing industry. Several tools useful for dealing with sequence information are introduced through the Biology Workbench.Information about sequence, structure, and active sites for many proteins is accessible online. This real world application of bioinformatics introduces us to amylases and asks us to explore functionally similar enzymes from markedly divergent organisms. Here, we search for microbial enzymes useful in the starch processing industry. Several tools useful for dealing with sequence information are introduced through the Biology Workbench. * investigate the use of bioinformatics to identify potentially useful microbial amylases for industry
Salivary cholinesterase activity in children with organic and convential diets
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....
Componential profile and amylase inhibiting activity of phenolic compounds from Calendula officinalis L. leaves.
Olennikov, Daniil N; Kashchenko, Nina I
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
Componential Profile and Amylase Inhibiting Activity of Phenolic Compounds from Calendula officinalis L. Leaves
Olennikov, Daniil N.; Kashchenko, Nina I.
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
Calcium-mediated conversion of sucrose to starch in relation to the activities of amylases and sucrose-metabolizing enzymes in sorghum grains raised through liquid culture.
Bhatia, S; Singh, R
Detached ears of sorghum (Sorghum vulgare) were cultured in complete liquid medium containing Ca2+(0, 3, 10 and 30 mM) and effect of this ion on the conversion of sucrose to starch with respect to the activities of amylases, sucrose synthase, sucrose phosphate synthase and soluble invertases were studied in developing grains. Presence of 3 mM Ca2+ in culture medium enhanced both accumulation of starch and activity of alpha-amylase in grain but without having any influence on the activity of beta-amylase. However, with 10 and 30 mM Ca2+, the accumulation of starch and activities of both amylases decreased and with advancement in culturing period, starch accumulation was further decreased. Irrespective of its concentration, Ca2+ enhanced the activities of sucrose synthase (synthesis), sucrose-phosphate synthase, soluble acid invertase and soluble-neutral invertase. Increase in the concentration of Ca2+ in culture medium was concomitant with an elevation in relative proportion of sucrose in the grain reflecting a net balance in per cent increase with Ca2+ in the activities of sucrose-synthesizing enzymes over sucrose-hydrolysing ones. Based on the results, it is suggested that assimilation of Ca2+ by grain is essential for maintaining high activity of alpha-amylase to generate starch primers required for the conversion of sucrose to starch during grain filling in sorghum. PMID:10983425
MDM2 Is Required for Suppression of Apoptosis by Activated Akt1 in Salivary Acinar Cells
Kirsten H. Limesand; Kathryn L. Schwertfeger; Steven M. Anderson
Chronic damage to the salivary glands is a common side effect following head and neck irradiation. It is hypothesized that irreversible damage to the salivary glands occurs immediately after radiation; however, previous studies with rat models have not shown a causal role for apoptosis in radiation-induced injury. We report that etoposide and gamma irradiation induce apoptosis of salivary acinar cells
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...
Association of Novel Domain in Active Site of Archaic Hyperthermophilic Maltogenic Amylase from Staphylothermus marinus*
Jung, Tae-Yang; Li, Dan; Park, Jong-Tae; Yoon, Se-Mi; Tran, Phuong Lan; Oh, Byung-Ha; Jane?ek, Štefan; Park, Sung Goo; Woo, Eui-Jeon; Park, Kwan-Hwa
Staphylothermus marinus maltogenic amylase (SMMA) is a novel extreme thermophile maltogenic amylase with an optimal temperature of 100 °C, which hydrolyzes ?-(1–4)-glycosyl linkages in cyclodextrins and in linear malto-oligosaccharides. This enzyme has a long N-terminal extension that is conserved among archaic hyperthermophilic amylases but is not found in other hydrolyzing enzymes from the glycoside hydrolase 13 family. The SMMA crystal structure revealed that the N-terminal extension forms an N? domain that is similar to carbohydrate-binding module 48, with the strand-loop-strand region forming a part of the substrate binding pocket with several aromatic residues, including Phe-95, Phe-96, and Tyr-99. A structural comparison with conventional cyclodextrin-hydrolyzing enzymes revealed a striking resemblance between the SMMA N? domain position and the dimeric N domain position in bacterial enzymes. This result suggests that extremophilic archaea that live at high temperatures may have adopted a novel domain arrangement that combines all of the substrate binding components within a monomeric subunit. The SMMA structure provides a molecular basis for the functional properties that are unique to hyperthermophile maltogenic amylases from archaea and that distinguish SMMA from moderate thermophilic or mesophilic bacterial enzymes. PMID:22223643
Differential RNA Expression of Two Barley ß-Amylase Genes (Bmy1 and Bmy2) in Developing Grains and Their Association with ß-Amylase Activity
Technology Transfer Automated Retrieval System (TEKTRAN)
RNA expression from the barley ß-amylase1 (Bmy1) gene was determined during seed development in four genotypes (Legacy, Harrington, Ashqelon, and PI 296897). The Bmy1 transcript amount in Legacy and Harrington was not significantly different at 17, 19, or 21 days after anthesis (DAA). Ashqelon Bmy...
Structural and functional aspects of chloride binding to Alteromonas haloplanctis alpha-amylase.
Feller, G; Bussy, O; Houssier, C; Gerday, C
Chloride is the allosteric effector of vertebrate pancreatic and salivary alpha-amylases and of the bacterial alpha-amylase from Alteromonas haloplanctis. Activation experiments of A. haloplanctis alpha-amylase by several monovalent anions show that a negative charge, not restricted to that of Cl-, is essential for the amylolytic reaction. Engineering of the chloride binding site reveals that a basic residue is an essential component of the site. The mutation K337R alters the Cl--binding properties, whereas the mutation K337Q produces an active, chloride-independent enzyme. Comparison of the Kd values for Cl- in three homologous alpha-amylases also indicates that the binding affinity is dependent on the chloride coordination mode by this basic residue. Analysis of substrate and chloride binding according to the allosteric kinetic model shows that the chloride effector is not involved in substrate binding. By contrast, the pH dependence of activity and experiments of chemical modifications and Ca2+ inhibition show that the chloride ion is responsible for the pKa shift of catalytic groups and interacts with active site carboxyl groups. PMID:8798613
Periodontal status, salivary immunoglobulin, and microbial counts after short exposure to an isolated environment.
Rai, Balwant; Kaur, Jasdeep
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
Age-independent increases in male salivary testosterone during horticultural activity among Tsimane forager-farmers
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
Age-independent increases in male salivary testosterone during horticultural activity among Tsimane forager-farmers.
Trumble, Benjamin C; Cummings, Daniel K; O'Connor, Kathleen A; Holman, Darryl J; Smith, Eric A; Kaplan, Hillard S; Gurven, Michael D
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
Heat shock inhibits. alpha. -amylase synthesis in barley aleurone without inhibiting the activity of endoplasmic reticulum marker enzymes
Sticher, L.; Biswas, A.K.; Bush, D.S.; Jones, R.L. (Univ. of California, Berkeley (USA))
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.
Effect of duct obstruction on histology and on activities of ?-glutamyl transferase, adenosine triphosphatase, alkaline phosphatase, and amylase in rat pancreas
Alan J. Graves; Doris R. G. Holmquist; Sherwood Githens
The effect of pancreatic duct obstruction on the activities of amylase and three nonexocrine pancreatic enzymes was studied in the rat. ?-Glutamyl transferase (GGTase) activity, which is localized primarily in the plasma membrane of acinar cells, disappeared from the acinar basolateral plasma membrane and declined in specific activity by 80% over a seven-day experimental period. Mg-ATPase, localized primarily in the
Effect of starch and amylase on the expression of amylase-binding protein A in Streptococcus gordonii
Nikitkova, A.E.; Haase, E.M.; Scannapieco, F.A.
SUMMARY Streptococcus gordonii is a common oral commensal bacterial species in tooth biofilm (dental plaque) and specifically binds to salivary amylase through the surface exposed amylase-binding protein A (AbpA). When S. gordonii cells are pretreated with amylase, amylase bound to AbpA facilitates growth with starch as a primary nutrition source. The goal of this study was to explore possible regulatory effects of starch, starch metabolites and amylase on the expression of S. gordonii AbpA. An amylase ligand-binding assay was used to assess the expression of AbpA in culture supernatants and on bacterial cells from S. gordonii grown in defined medium supplemented with 1% starch, 0.5 mg ml?1 amylase, with starch and amylase together, or with various linear malto-oligosaccharides. Transcription of abpA was determined by reverse transcription quantitative polymerase chain reaction. AbpA was not detectable in culture supernatants containing either starch alone or amylase alone. In contrast, the amount of AbpA was notably increased when starch and amylase were both present in the medium. The expression of abpA was significantly increased (P < 0.05) following 40 min of incubation in defined medium supplemented with starch and amylase. Similar results were obtained in the presence of maltose and other short-chain malto-oligosacchrides. These results suggest that the products of starch hydrolysis produced from the action of salivary ?-amylase, particularly maltose and maltotriose, regulate AbpA expression in S. gordonii. PMID:22759313
Effect of starch and amylase on the expression of amylase-binding protein A in Streptococcus gordonii.
Nikitkova, A E; Haase, E M; Scannapieco, F A
Streptococcus gordonii is a common oral commensal bacterial species in tooth biofilm (dental plaque) and specifically binds to salivary amylase through the surface exposed amylase-binding protein A (AbpA). When S. gordonii cells are pretreated with amylase, amylase bound to AbpA facilitates growth with starch as a primary nutrition source. The goal of this study was to explore possible regulatory effects of starch, starch metabolites and amylase on the expression of S. gordonii AbpA. An amylase ligand-binding assay was used to assess the expression of AbpA in culture supernatants and on bacterial cells from S. gordonii grown in defined medium supplemented with 1% starch, 0.5 mg ml(-1) amylase, with starch and amylase together, or with various linear malto-oligosaccharides. Transcription of abpA was determined by reverse transcription quantitative polymerase chain reaction. AbpA was not detectable in culture supernatants containing either starch alone or amylase alone. In contrast, the amount of AbpA was notably increased when starch and amylase were both present in the medium. The expression of abpA was significantly increased (P < 0.05) following 40 min of incubation in defined medium supplemented with starch and amylase. Similar results were obtained in the presence of maltose and other short-chain malto-oligosacchrides. These results suggest that the products of starch hydrolysis produced from the action of salivary ?-amylase, particularly maltose and maltotriose, up-regulate AbpA expression in S. gordonii. PMID:22759313
... salivary gland tumors usually show up as painless enlargements of these glands. Tumors rarely involve more than ... otolaryngologist-head and neck surgeon should check these enlargements. Malignant tumors of the major salivary glands can ...
Effects of sorghum (Sorghum bicolor (L.) Moench) tannins on ?-amylase activity and in vitro digestibility of starch in raw and processed flours.
Mkandawire, Nyambe L; Kaufman, Rhett C; Bean, Scott R; Weller, Curtis L; Jackson, David S; Rose, Devin J
The purpose of this study was to investigate the effects of tannins on starch digestion in tannin-containing sorghum extracts and wholegrain flours from 12 sorghum varieties. Extracts reduced amylase activity in a tannin concentration-dependent manner when the extract was mixed with the enzyme before substrate (amylopectin) addition, with higher molecular weight tannins showing greater reduction. Conversely, when the extract and substrate were combined before enzyme addition an enhancement in amylase activity was experienced. In uncooked, cooked, and cooked and stored wholegrain sorghum flours, rapidly digestible, slowly digestible, and resistant starches were not correlated with tannin content or molecular weight distribution. Resistant starch increased from 6.5% to 22-26% when tannins were added to starch up to 50% (starch weight). Tannin extracts both reduced and enhanced amylase activity depending on conditions, and, while these trends were clear in extracts, the effects on starch digestion in wholegrain flours was more complex. PMID:23581620
A new nano-optical sensor thin film cadmium sulfide doped in sol-gel matrix for assessment of ?-amylase activity in human saliva.
Attia, M S; Zoulghena, H; Abdel-Mottaleb, M S A
A novel, simple, sensitive and precise spectrofluorimetric method is developed for measuring the activity of the ?-amylase enzyme in human saliva. The remarkable quenching of the luminescence intensity at 634 nm of nano CdS doped in a sol-gel matrix by various concentrations of maltose (produced from the reaction of the enzyme with the starch substrate) was successfully used as an optical sensor for the assessment of ?-amylase activity. The calibration plot was achieved over the concentration range 4.8 × 10(-10) to 1.2 × 10(-5) mol L(-1) maltose with a correlation coefficient of 0.999 and a detection limit of 5.7 × 10(-11) mol L(-1). The method was used satisfactorily for assessment of the ?-amylase activity in a number of human saliva samples collected from various healthy volunteers. PMID:24358458
Asparagine as a nitrogen source for improving the secretion of mouse alpha-amylase in Saccharomyces cerevisiae protease A-deficient strains.
Chen, D C; Wang, B D; Chou, P Y; Kuo, T T
A modified chemically defined medium was achieved by using asparagine as a nitrogen source to increase the production of secreted mouse alpha-amylase in several Saccharomyces cerevisiae protease A-deficient (pep4) strains. The specific productivity (quantity) and the 53 kDa non-glycosylated active form (quality) of mouse salivary alpha-amylase in liquid medium containing asparagine was remarkably improved compared to media containing other nitrogen sources, including ammonium sulphate, glutamic acid, arginine, casamino acids, yeast extract and peptone. Similar improvement was also observed on starch solid agar regarding the clarity and size of the halo zone formed by alpha-amylase activity. Compared with ammonium sulphate, advantages of using asparagine as the nitrogen source in liquid or solid medium included increasing the cell mass of test strains, recovering the viability of protease-deficient strains to levels similar to the wild-type strain, and increasing the copy number of the mouse alpha-amylase expression vector in test strains. In turn, these advantages apparently contributed to the increase of secretion of mouse alpha-amylase in several test strains and especially in the protease A-deficient strains. In addition to demonstrating the use of modified chemically defined medium to improve the quality and quantity of secreted mouse alpha-amylase, this study also provides a new strategy to improve the secretion of heterologous proteins in protease A deficient strains. PMID:10649450
Original article Amylase polymorphism
Boyer, Edmond
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
Effect of tin oxide nanoparticle binding on the structure and activity of ?-amylase from Bacillus amyloliquefaciens
NASA Astrophysics Data System (ADS)
Jahir Khan, Mohammad; Qayyum, Shariq; Alam, Fahad; Husain, Qayyum
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.
Effect of tin oxide nanoparticle binding on the structure and activity of ?-amylase from Bacillus amyloliquefaciens.
Khan, Mohammad Jahir; Qayyum, Shariq; Alam, Fahad; Husain, Qayyum
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 (K(m)), was found to be 26.96 and 28.45 mg ml(-1), while V(max) was 4.173 and 3.116 mg ml(-1) min(-1) for free and NP-bound enzyme, respectively. PMID:22020314
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.
Ahmed, Dildar; Mughal, Qaria Mumtaz; Younas, Saba; Ikram, Muhammad
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
Purification of ?-amylases using magnetic alginate beads
Sunita Teotia; M. N. Gupta
Magnetic alginate beads were used to purify ?-amylases from porcine pancreas, starchzyme, BAN 240L (a commercial purification\\u000a from Bacillus subtilis), and wheat germ. The beads bound a significant level of ?-amylase activity from porcine pancreas, BAN 240L, and wheat germ.\\u000a In each case, the enzyme activity could be eluted by using 1.0 M maltose, a known competitive inhibitor of ?-amylase.
Gamma irradiation of sorghum flour: Effects on microbial inactivation, amylase activity, fermentability, viscosity and starch granule structure
NASA Astrophysics Data System (ADS)
Mukisa, Ivan M.; Muyanja, Charles M. B. K.; Byaruhanga, Yusuf B.; Schüller, Reidar B.; Langsrud, Thor; Narvhus, Judith A.
Malted and un-malted sorghum ( Sorghum bicolor (L.) Moench) flour was gamma irradiated with a dose of 10 kGy and then re-irradiated with 25 kGy. The effects of irradiation on microbial decontamination, amylase activity, fermentability (using an amylolytic L. plantarum MNC 21 strain), starch granule structure and viscosity were determined. Standard methods were used during determinations. The 10 kGy dose had no effect on microbial load of un-malted flour but reduced that of malted flour by 3 log cycles. Re-irradiation resulted in complete decontamination. Irradiation of malt caused a significant ( p<0.05) reduction in alpha and beta amylase activity (22% and 32%, respectively). Irradiation of un-malted flour increased the rates of utilization of glucose and maltose by 53% and 100%, respectively, during fermentation. However, microbial growth, rate of lactic acid production, final lactic acid concentration and pH were not affected. Starch granules appeared normal externally even after re-irradiation, however, granules ruptured and dissolved easily after hydration and gelatinization. Production of high dry matter density porridge (200 g dry matter/L) with a viscosity of 3500 cP was achieved by irradiation of un-malted flout at 10 kGy. Gamma irradiation can be used to decontaminate flours and could be utilized to produce weaning porridge from sorghum.
Differential RNA Expression of ßm1 during Late Seed Development in Cultivated and Wild Barleys Carrying Different ßmy1 Intron III Alleles and the Association with Beta-Amylase Activity
Technology Transfer Automated Retrieval System (TEKTRAN)
Four genotypes carrying different beta-amylase 1 (Bmy1) intron III alleles (Bmy1.a, Bmy1.b, Bmy1.c, and Bmy1.d) were analyzed for differences in Bmy1 mRNA accumulation, beta-amylase activity and protein, and total protein during late seed development. Wild barleys (Hordeum vulgare ssp. spontaneum) ...
Galanin: hydrokinetic action on salivary glands in man.
Bauer, F E; Ghatei, M A; Zintel, A; Bloom, S R
Galanin was infused intravenously into eight healthy volunteers at a dose of 40 pmol kg-1 min-1 for 1 h to investigate the pharmacological effects of this peptide on the postprandial sialagogical response in man. Galanin significantly increased the salivary volume and the saliva output of sodium, chloride and bicarbonate compared to control saline infusion, but had no effect on the output of potassium and alpha-amylase. An increase in salivary pH was also observed. The increase in salivary volume may indicate a physiological role of galanin in the control of salivary secretion. PMID:2485092
Thermal adaptation of ?-amylases: a review.
Hiteshi, Kalpana; Gupta, Reena
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
A novel cold-active and salt-tolerant ?-amylase from marine bacterium Zunongwangia profunda: molecular cloning, heterologous expression and biochemical characterization.
Qin, Yongjun; Huang, Zongqing; Liu, Ziduo
A novel gene (amyZ) encoding a cold-active and salt-tolerant ?-amylase (AmyZ) was cloned from marine bacterium Zunongwangia profunda (MCCC 1A01486) and the protein was expressed in Escherichia coli. The gene has a length of 1785 bp and encodes an ?-amylase of 594 amino acids with an estimated molecular mass of 66 kDa by SDS-PAGE. The enzyme belongs to glycoside hydrolase family 13 and shows the highest identity (25%) to the characterized ?-amylase TVA II from thermoactinomyces vulgaris R-47. The recombinant ?-amylase showed the maximum activity at 35 °C and pH 7.0, and retained about 39% activity at 0 °C. AmyZ displayed extreme salt tolerance, with the highest activity at 1.5 M NaCl and 93% activity even at 4 M NaCl. The catalytic efficiency (k cat/K m) of AmyZ increased from 115.51 (with 0 M NaCl) to 143.30 ml mg(-1) s(-1) (with 1.5 M NaCl) at 35 °C and pH 7.0, using soluble starch as substrate. Besides, the thermostability of the enzyme was significantly improved in the presence of 1.5 M NaCl or 1 mM CaCl2. AmyZ is one of the very few ?-amylases that tolerate both high salinity and low temperatures, making it a potential candidate for research in basic and applied biology. PMID:24318109
Dietary effects of harmine, a ?-carboline alkaloid, on development, energy reserves and ?-amylase activity of Plodia interpunctella Hübner (Lepidoptera: Pyralidae).
Bouayad, Noureddin; Rharrabe, Kacem; Lamhamdi, Mostafa; Nourouti, Naima Ghailani; Sayah, Fouad
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
Lundep, a Sand Fly Salivary Endonuclease Increases Leishmania Parasite Survival in Neutrophils and Inhibits XIIa Contact Activation in Human Plasma
Chagas, Andrezza C.; Oliveira, Fabiano; Debrabant, Alain; Valenzuela, Jesus G.; Ribeiro, José M. C.; Calvo, Eric
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
Lubrication of selected salivary molecules and artificial salivas.
Aguirre, A; Mendoza, B; Reddy, M S; Scannapieco, F A; Levine, M J; Hatton, M N
The lubrication regime displayed by human salivas (parotid and submandibular-sublingual), purified salivary molecules (the mucins MG1 and MG2 and alpha-amylases), and selected artificial salivas (Oracare D, Saliva Substitute, and Orthana) was assessed in vitro using a friction-testing device. Thin-film (boundary) lubrication was observed for all of the salivary samples and two of the artificial salivas examined. Oracare D, a glycerol-based artificial saliva, was the exception since it lubricated by a thick-film (hydrodynamic) regime. On a molar basis, the best lubricants of the purified salivary molecules were MG1 greater than MG2 approximately nonglycosylated alpha-amylases approximately glycosylated alpha-amylases. PMID:2484182
[Construction of Pichia pastoris strain expressing salivary plasminogen activator from vampire bat (Desmodus rotundus)].
Liu, Yan; Su, Chang; Song, Xiaoshuang; Tang, Yalan; Bao, Zhenhong
Vampire bat saliva contains a plasminogen activator that presumably assists these hematophagous animals during feeding. Bat-PA (H), the full-length form of Vampire Bat Salivary Plasminogen Activator (DSPAalpha1), is homologous and similar efficacy to tissue-type plasminogen activator (t-PA). The strict fibrin dependence of activity is a characteristic which could be desirable in the fibrinolytic therapy. It is a unique fibrinolytic enzyme that does not promote neurodegeneration. In this study, according to the reported gene sequence (GenBank Accession No. J05082) of Vampire bat (D. rotundus) plasminogen activator. It was the first time to synthesize the full sequence of DSPAalpha1 in vitro and clone it into the expression vector pPIC9K, the recombinant plasmid was linearized and transformed into Pichia pastoris GS115 strain. Secreted expression of recombinant DSPAalpha1 was attained by methanol induction and its molecular mass is 47 kD. To get recombinant GS115 with high amount of protein, hundreds of His+ transformants had been screened to isolate clones resistant to high levels G418 (2-4 mg/mL), the selected clones mini-expressed in Pichia pastoris, and tested their fibrinolytic activities and expressed protein bands by fibrin plate assay and SDS-PAGE. DSPAalpha1 was determined by optical density after SDS-PAGE, the yield is about 30 mg per liter of fermentation culture. DSPAalpha1 derived often from mammalian cells: Chinese hamster ovary (CHO) cells, Baby hamster kidney (BHK) cells, COS cells, which might be produced at high cost. In Pichia pastoris, it is expected to higher yield and lower cost, thus it might be able to serve as new thrombolytic candidate. PMID:19637633
Thermally Stable Amylases from Antarctic Psychrophilic Bacteria
Technology Transfer Automated Retrieval System (TEKTRAN)
Hydrolysis of starch in cold environments by psychrophilic species of bacteria is believed to be accomplished through the production of special cold-adapted amylases. These amylases are reportedly thermally labile with low (<40 deg C) temperature optima and high specific activities at 0 to 25 deg C....
Original article Effect of age and exogenous amylase
Paris-Sud XI, Université de
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
Chemical modification of wheat ?-amylase by trinitrobenzenesulfonic acid, methoxypolyethylene glycol, and glutaraldehyde to improve its thermal stability and activity.
Daba, Tadessa; Kojima, Kenji; Inouye, Kuniyo
The amino groups of wheat ?-amylase (WBA) were modified by 2,4,6-trinitrobenzenesulfonic acid (TNBS), 2,4-bis (O-methoxypolyethylene glycol)-6-chloro-s-triazine (mPEG), and glutaraldehyde (GA) to improve its thermal stability and activity. Modification of WBA by 5mM TNBS, 4.8?M mPEG and 11 mM GA improved its T50 (the temperature at which 50% of its activity is lost after 30 min of incubation) from 47 ± 1°C to 48 ± 2, 55 ± 2, and 54 ± 2°C, respectively. The catalytic activity of WBA was reduced by 15% and 59% with modification by 5mM TNBS and 11mM GA, respectively. In all cases, the enhancement of thermostability of modified WBA was entropically driven. The activity of WBA modified by 4.8?M mPEG was enhanced by 39% at 25°C. Therefore, the thermal stability of WBA was significantly improved by modification with mPEG, GA and slightly by TNBS and its catalytic activity was enhanced by mPEG. PMID:24315646
Effects of infrared radiation, solar cooking and microwave cooking on alpha-amylase inhibitor in sorghum (Sorghum bicolor L.).
Mulimani, V H; Supriya, D
Three domestic cooking methods were studied in alpha-amylase inhibitory activity in sorghum grains. In all the treatments, overnight soaked seeds lost amylase inhibitory activity much faster. All the three treatments reduced the inhibitory activity. Use of solar cooker for reducing amylase inhibitory activity works out very economically and efficiently. Microwave cooking eliminates amylase inhibitory activity within 5 minutes. PMID:7855094
Effect of tannic acid-fish scale gelatin hydrolysate hybrid nanoparticles on intestinal barrier function and ?-amylase activity.
Wu, Shao-Jung; Ho, Yi-Cheng; Jiang, Shun-Zhou; Mi, Fwu-Long
Practical application of tannic acid is limited because it readily binds proteins to form insoluble aggregates. In this study, tannic acid was self-assembled with fish scale gelatin hydrolysates (FSGH) to form stable colloidal complex nanoparticles. The nanoparticles prepared from 4 mg ml(-1) tannic acid and 4 mg ml(-1) FSGH had a mean particle size of 260.8 ± 3.6 nm, and showed a positive zeta potential (20.4 ± 0.4 mV). The nanoparticles acted as effective nano-biochelators and free radical scavengers because they provided a large number of adsorption sites for interaction with heavy metal ions and scavenging free radicals. The maximum adsorption capacity for Cu(2+) ions was 123.5 mg g(-1) and EC50 of DPPH radical scavenging activity was 21.6 ± 1.2 ?g ml(-1). Hydroxyl radical scavenging effects of the nanoparticles were investigated by electron spin resonance spectroscopy. The copper-chelating capacity and free radical scavenging activity of the nanoparticles were associated with their capacity to inhibit Cu(2+) ion-induced barrier impairment and hyperpermeability of Caco-2 intestinal epithelial tight junction (TJ). However, ?-amylase inhibitory activity of the nanoparticles was significantly lower than that of free tannic acid. The results suggest that the nanoparticles can ameliorate Cu(2+) ion induced intestinal epithelial TJ dysfunction without severely inhibiting the activity of the digestive enzymes. PMID:26069899
Profiles and ?-amylase inhibition activity of proanthocyanidins in unripe Manilkara zapota (chiku).
Wang, Hongyu; Liu, Tingting; Song, Lixia; Huang, Dejian
Proanthocyanidins in unripe Manilkara zapota (chiku) were isolated using solvent extraction followed by Sephadex LH-20 fractionation with a yield of 0.9%. HPLC analysis using a diol column revealed well-resolved oligomers ranging from dimer to hexamer. The majority of the proanthocyanidins are composed of higher-degree oligomers appearing as one large peak in the chromatogram. Analysis of the proanthocyanidins using LC/MS showed that (epi)gallocatechins were the dominant extension unit in the proanthocyanidins. In agreement with this result, thiolysis treatment of the proanthocyanidins using mercaptoacetic acid produced thioether derivatives of (epi)gallocatechins as the major product and (epi)gallocatechin gallate derivatives as the minor product. The mean of the degree of polymerization was estimated to be 9.0. From MALDI-TOF MS, B-type gallocatechin oligomers up to decamer could be detected. The unripe chiku proanthocyanidins are thus good starting material for preparation of (epi)gallocatechin derivatives. The proanthocyanidins was shown to inhibit ?-amylase with an IC(50) value of 4.2 ± 0.2 ?g/mL and inhibit ?-glucosidase with an IC(50) of 16.6 ± 0.3 ?g/mL. PMID:22394060
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
Cochran-Stafira, D. Liane
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
Salivary gland acinar cells regenerate functional glandular structures in modified hydrogels
NASA Astrophysics Data System (ADS)
Pradhan, Swati
Xerostomia, a condition resulting from irradiation of the head and neck, affects over 40,000 cancer patients each year in the United States. Direct radiation damage of the acinar cells that secrete fluid and protein results in salivary gland hypofunction. Present medical management for xerostomia for patients treated for upper respiratory cancer is largely ineffective. Patients who have survived their terminal diagnosis are often left with a diminished quality of life and are unable to enjoy the simple pleasures of eating and drinking. This project aims to ultimately reduce human suffering by developing a functional implantable artificial salivary gland. The goal was to create an extracellular matrix (ECM) modified hyaluronic acid (HA) based hydrogel culture system that allows for the growth and differentiation of salivary acinar cells into functional acini-like structures capable of secreting large amounts of protein and fluid unidirectionally and to ultimately engineer a functional artificial salivary gland that can be implanted into an animal model. A tissue collection protocol was established and salivary gland tissue was obtained from patients undergoing head and neck surgery. The tissue specimen was assessed by histology and immunohistochemistry to establish the phenotype of normal salivary gland cells including the native basement membranes. Hematoxylin and eosin staining confirmed normal glandular tissue structures including intercalated ducts, striated ducts and acini. alpha-Amylase and periodic acid schiff stain, used for structures with a high proportion of carbohydrate macromolecules, preferentially stained acinar cells in the tissue. Intercalated and striated duct structures were identified using cytokeratins 19 and 7 staining. Myoepithelial cells positive for cytokeratin 14 were found wrapped around the serous and mucous acini. Tight junction components including ZO-1 and E-cadherin were present between both ductal and acinar cells. Ductal and acinar cells were identified in cultured cells from dispersed tissue. Biomarker studies with the salivary enzyme, alpha-amylase, and tight junction proteins, such as zonula occludens-1 and E-cadherin, confirmed the phenotype of these cells. Strong staining for laminin and perlecan/HSPG2 were noted in basement membranes and perlecan also was secreted and organized by cultured acinar populations, which formed lobular structures that mimicked intact glands when cultured on Matrigel(TM) or a bioactive peptide derived from domain IV of perlecan (PlnDIV). On either matrix, large acini-like lobular structures grew and formed connections between the lobes. alpha-Amylase secretion was confirmed by staining and activity assay. Biomarkers including tight junction protein E-cadherin and water channel protein, aquaporin 5 (AQP5) found in tissue, were expressed in cultured acinar cells. Cells cultured on Matrigel(TM) or PlnDIV peptide organized stress fibers and activated focal adhesion kinase (FAK). HA, a natural polysaccharide and a major component of the ECM, can be used to generate soft and pliable hydrogels. A culture system consisting of HA hydrogel and PlnDIV peptide was used to generate a 2.5D culture system. Acinar cells cultured on these hydrogels self-assembled into lobular structures and expressed tight junction components such as ZO-1. Acini-like structures were stained for the presence of alpha-amylase. Live/dead staining revealed the presence of apoptotic cells in the center of the acini-like structures, indicative of lumen formation. The functionality of these acini-like structures was studied by stimulating them with neurotransmitters to enhance their fluid and protein production. Acini-like structures treated with norepinephrine and isoproterenol showed increased granule formation as observed by phase contrast microscopy and alpha-amylase staining in the structures. Lobular structures on hydrogels were treated with acetylcholine to increase fluid production. The increase in intracellular calcium due to the activation of the M3 muscarinic receptor via binding to ac
A study into salivary-based measurement of human stress subjected to ellestad stress test protocol
Y. K. Lee; A. Za'aba; N. K. Madzhi; A. Ahmad
Previous works on the effects of salivary alpha amylase in respond to various stressors report encouraging findings on it being a good indicator of stress. Ellestad protocol is a clinical procedure to screen for coronary artery disease by introducing exercise induced physical stress. If a salivary based biomarker profile in accordance to a stress test protocol could be established, the
General Biochemical Characterization of Thermostable Extracellular ?-Amylase from Clostridium thermosulfurogenes
Hyun, H. H.; Zeikus, J. G.
Clostridium thermosulfurogenes, an anaerobic bacterium which ferments starch into ethanol at 62°C, produced an active extracellular amylase and contained intracellular glucoamylase but not pullulanase activity. The extracellular amylase was purified 2.4-fold, and its general physicochemical and catalytic properties were examined. The extracellular amylase was characterized as a ?-amylase (1,4-?-d-glucan maltohydrolase) based on demonstration of exocleavage activity and the production of maltose with a ?-anomeric configuration from starch. The ?-amylase activity was stable and optimally active at 80 and 75°C, respectively. The pH optimum for activity and the pH stability range was 5.5 to 6 and 3.5 to 6.5, respectively. The apparent [S]0.5V and Vmax for ?-amylase activity on starch was 1 mg/ml and 60 U/mg of protein. Similar to described ?-amylase, the enzyme was inhibited by p-chloromercuribenzoate, Cu2+, and Hg2+; however, ?- and ?-cyclodextrins were not competitive inhibitors. The ?-amylase was active and stable in the presence of air or 10% (vol/vol) ethanol. The ?-amylase and glucoamylase activities enabled the organism to actively ferment raw starch in the absence of significant pullulanase or ?-amylase activity. PMID:16346789
Expression and Localization of ?-amylase in the Submandibular and Sublingual Glands of Mice
Yamagishi, Ryoko; Wakayama, Tomohiko; Nakata, Hiroki; Adthapanyawanich, Kannika; Kumchantuek, Tewarat; Yamamoto, Miyuki; Iseki, Shoichi
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
Cortisol and Children's Adjustment: The Moderating Role of Sympathetic Nervous System Activity
ERIC Educational Resources Information Center
El-Sheikh, Mona; Erath, Stephen A.; Buckhalt, Joseph A.; Granger, Douglas A.; Mize, Jacquelyn
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…
Candida albicans Cek1 Mitogen-Activated Protein Kinase Signaling Enhances Fungicidal Activity of Salivary Histatin 5.
Li, Rui; Puri, Sumant; Tati, Swetha; Cullen, Paul J; Edgerton, Mira
Candida albicans is a major etiological organism for oropharyngeal candidiasis (OPC), while salivary histatin 5 (Hst 5) is a human fungicidal protein that protects the oral cavity from OPC. C. albicans senses its environment by mitogen-activated protein kinase (MAPK) activation that can also modulate the activity of some antifungal drugs, including Hst 5. We found that phosphorylation of the MAPK Cek1, induced either by N-acetylglucosamine (GlcNAc) or serum, or its constitutive activation by deletion of its phosphatase Cpp1 elevated the susceptibility of C. albicans cells to Hst 5. Cek1 phosphorylation but not hyphal formation was needed for increased Hst 5 sensitivity. Interference with the Cek1 pathway by deletion of its head sensor proteins, Msb2 and Sho1, or by addition of secreted aspartyl protease (SAP) cleavage inhibitors, such as pepstatin A, reduced Hst 5 susceptibility under Cek1-inducing conditions. Changes in fungal cell surface glycostructures also modulated Hst 5 sensitivity, and Cek1-inducing conditions resulted in a higher uptake rate of Hst 5. These results show that there is a consistent relationship between activation of Cek1 MAPK and increased Hst 5 susceptibility in C. albicans. PMID:25824232
The brown seaweed Sargassum hemiphyllum exhibits ?-amylase and ?-glucosidase inhibitory activity and enhances insulin release in vitro.
Hwang, Pai-An; Hung, Yu-Lan; Tsai, Yi-Kuan; Chien, Shih-Yung; Kong, Zwe-Ling
Diabetes is one of the most prevalent chronic diseases globally. In this study, major polyphenols (17.35 ± 0.93-36.66 ± 2.01 mg/g) and minor fucoxanthin (non detected 15.12 ± 0.09 mg/g) were isolated from water, ethanol, and acetone extracts (WES, EES, and AES, respectively) of Sargassum hemiphyllum. Inhibition of ?-amylase, ?-glucosidase, sucrose, and maltase activities and stimulation of insulin secretion was greater with AES than with WES or EES and correlated with polyphenol and fucoxanthin concentrations in extracts. Moreover, 250 ?g/ml EES and AES significantly increased insulin secretion in the presence of 25 mg/ml glibenclamide to higher levels than those obtained with 50 mg/ml glibenclamide. None of the extracts exhibited cytotoxicity, exacerbated the side effects of glibenclamide, or inhibited glibenclamide-induced insulin secretion. These results suggested that the S. hemiphyllum extracts WES, EES, and AES could be used as pharmaceuticals and functional foods to reduce dosages of synthetic diabetes drugs. PMID:25344877
Salivary Surrogates of Plasma Nitrite and Catecholamines during a 21-Week Training Season in Swimmers
Díaz Gómez, Miguel Mauricio; Bocanegra Jaramillo, Olga Lucia; Teixeira, Renata Roland; Espindola, Foued Salmen
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
Novel Family of Insect Salivary Inhibitors Blocks Contact Pathway Activation by Binding to Polyphosphate, Heparin, and Dextran Sulfate
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.
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
Cyclic changes in salivary activity of N-acetyl-beta-d-glucosaminidase; a possible efficient indicator for predicting ovulation and pregnancy.
Rosado, A; Delgado, N M; Valázquez, A; Aznar, R; Martínez-Manautou, J
Salivary activity of N-acetyl-beta-D-glucosaminidase showed a characteristic pattern of changes during the normal menstrual cycle with a distinct peak on Day 13, 14, or 15 before the next menstruation. This peak of enzyme activity occurred withing one day of the nadir of basal body temperature and was absent in women with spontaneous or iatrogenic anovulatory cycles. These results are stongly suggestive that the salivary determination of this activity may be convenient indicatior for determining the day of ovulation. PMID:879215
Characterization of the native form and the carboxy-terminally truncated halotolerant form of ?-amylases from Bacillus subtilis strain FP-133.
Takenaka, Shinji; Miyatake, Ayaka; Tanaka, Kosei; Kuntiya, Ampin; Techapun, Charin; Leksawasdi, Noppol; Seesuriyachan, Phisit; Chaiyaso, Thanongsak; Watanabe, Masanori; Yoshida, Ken-Ichi
Two amylases, amylase I and amylase II from Bacillus subtilis strain FP-133, were purified to homogeneity and characterized. Their stabilities toward temperature, pH, and organic solvents, and their substrate specificities toward polysaccharides and oligosaccharides were similar. Under moderately high salt conditions, both amylases were more stable than commercial B. licheniformis amylase, and amylase I retained higher amylase activity than amylase II. The N-terminal amino acid sequence, genomic southern blot analysis, and MALDI-TOFF-MS analysis indicated that the halotolerant amylase I was produced by limited carboxy-terminal truncation of the amylase II peptide. The deduced amino acid sequence of amylase II was >95% identical to that of previously reported B. subtilis ?-amylases, but their carboxy-terminal truncation points differed. Three recombinant amylases - full-length amylase corresponding to amylase II, an artificially truncated amylase corresponding to amylase I, and an amylase with a larger artificial C-terminal truncation - were expressed in B. subtilis. The artificially truncated recombinant amylases had the same high amylase activity as amylase I under moderately high salt conditions. Sequence comparisons indicated that an increased ratio of Asp/Glu residues in the enzyme may be one factor responsible for increasing halotolerance. PMID:25689045
Expression of beta-amylase from alfalfa taproots.
Gana, J A; Kalengamaliro, N E; Cunningham, S M; Volenec, J J
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
Bone-muscle unit activity, salivary steroid hormones profile, and physical effort over a 3-week stage race.
Grasso, D; Corsetti, R; Lanteri, P; Di Bernardo, C; Colombini, A; Graziani, R; Banfi, G; Lombardi, G
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
[Salivary glands secretory activity in patients with terminal chronic renal insufficiency on programmed dialysis].
Afanas'ev, V V; Vavilova, T P; Osokin, M V; Pushkina, A V
Saliva secretion speed and some biochemical mixed saliva characteristics were studied in patients with a terminal stage of chronic renal insufficiency. There was reduction of salivary function by more than 2 times and an increase of urea and creatinine concentrations in mixed saliva before the dialysis start. In case of higher urea content in saliva the speed of salivation was the highest that could evidence for an adequate response of the salivary glands to toxic action of nitrogen metabolism end products. The function restoration after hemodialysis took place not in all the patients. Amino acid catabolism product concentration in mixed saliva fell after hemodialysis and correlated directly with the amount of urea and creatinine in blood plasma. It took place also in "urea ricochet" when its content in blood increased sharply 1 hour after hemodialysis due to urea washout from the tissues. PMID:17159840
Cognitive Function and Salivary DHEA Levels in Physically Active Elderly African American Women.
Davis, Greggory R; Gallien, Gabrielle J; Moody, Kaitlyn M; LeBlanc, Nina R; Smoak, Peter R; Bellar, David
Serum and plasma dehydroepiandrosterone sulfate (DHEAS) concentration has been associated with several health parameters associated with aging including cognitive function, bone mineral density, and muscular strength. However, the effectiveness of salivary DHEA for the prediction of cognitive function, bone mineral density, and muscular strength in older adults is currently unknown. Thirty elderly African American females provided early morning salivary samples and DHEA levels were determined using a commercially available immunoassay. Participants completed testing for psychomotor and executive function via Trail Making Tests (TMT) A and B, respectively. Bone ultrasound attenuation (BUA) was used to bone density and an isometric mid-thigh pull (IMTP) was used to determine isometric strength. Age significantly correlated with time on TMT A (r=0.328) and B (r=0.615) but was not related to DHEA, BUA, or IMTP outcomes. Elevated DHEA was associated with longer time to completion for TMT A (? (2) = 5.14) but not to TMT B. DHEA levels were not associated with BUA or IMTP outcomes. While elevated levels of DHEA were correlated with impaired psychomotor function, salivary DHEA is not associated with executive function, bone mineral density, or isometric strength in elderly African American women. PMID:26064106
Cognitive Function and Salivary DHEA Levels in Physically Active Elderly African American Women
Gallien, Gabrielle J.; Moody, Kaitlyn M.; LeBlanc, Nina R.; Smoak, Peter R.; Bellar, David
Serum and plasma dehydroepiandrosterone sulfate (DHEAS) concentration has been associated with several health parameters associated with aging including cognitive function, bone mineral density, and muscular strength. However, the effectiveness of salivary DHEA for the prediction of cognitive function, bone mineral density, and muscular strength in older adults is currently unknown. Thirty elderly African American females provided early morning salivary samples and DHEA levels were determined using a commercially available immunoassay. Participants completed testing for psychomotor and executive function via Trail Making Tests (TMT) A and B, respectively. Bone ultrasound attenuation (BUA) was used to bone density and an isometric mid-thigh pull (IMTP) was used to determine isometric strength. Age significantly correlated with time on TMT A (r=0.328) and B (r=0.615) but was not related to DHEA, BUA, or IMTP outcomes. Elevated DHEA was associated with longer time to completion for TMT A (?2 = 5.14) but not to TMT B. DHEA levels were not associated with BUA or IMTP outcomes. While elevated levels of DHEA were correlated with impaired psychomotor function, salivary DHEA is not associated with executive function, bone mineral density, or isometric strength in elderly African American women.
Interferon-? Induces Immunoproteasomes and the Presentation of MHC I-Associated Peptides on Human Salivary Gland Cells
Arellano-Garcia, Martha E.; Misuno, Kaori; Tran, Simon D.; Hu, Shen
A prominent histopathological feature of Sjögren's syndrome, an autoimmune disease, is the presence of lymphocytic infiltrates in the salivary and lachrymal glands. Such infiltrates are comprised of activated lymphocytes and macrophages, and known to produce multiple cytokines including interferon-gamma (IFN-?). In this study, we have demonstrated that IFN-? strongly induces the expression of immunoproteasome beta subunits (?1i, ?2i and ?5i) and immunoproteasome activity but conversely inhibits the expression of proteasome beta subunits (?1, ?2 and ?5) in human salivary gland (HSG) cells. Mass spectrometric analysis has revealed potential MHC I-associated peptides on the HSG cells, including a tryptic peptide derived from salivary amylase, due to IFN-? stimulation. These results suggest that IFN-? induces immunoproteasomes in HSG cells, leading to enhanced presentation of MHC I-associated peptides on cell surface. These peptide-presenting salivary gland cells may be recognized and targeted by auto-reactive T lymphocytes. We have also found that lactacystin, a proteasome inhibitor, inhibits the expression of ?1 subunit in HSG cells and blocks the IFN-?-induced expression of ?1i and immunoproteasome activity. However, the expression of ?2i and ?5i in HSG cells is not affected by lactacystin. These results may add new insight into the mechanism regarding how lactacystin blocks the action of proteasomes or immunoproteasomes. PMID:25102056
Water Stress Enhances Expression of an ?-Amylase Gene in Barley Leaves
Jacobsen, John V.; Hanson, Andrew D.; Chandler, Peter C.
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
Inhibition of Sunn Pest, Eurygaster integriceps, ?-Amylases by ?-Amylase Inhibitors (T-?AI) from Triticale
Mehrabadi, Mohammad; Bandani, Ali R.; Saadati, Fatemeh
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
Wheat tetrameric inhibitors of insect ?-amylases: Alloploid heterosis at the molecular level
Gomez, Luis; Sanchez-Monge, Rosa; Garcia-Olmedo, Francisco; Salcedo, Gabriel
Tetrameric inhibitors of heterologous ?-amylases have been characterized in allohexaploid wheat, Triticum aestivum (genomes AABBDD), as well as in Triticum turgidum (AABB) and Triticum tauschii (DD). Their subunits have been identified as the previously described CM proteins. Single oligomeric species were observed in T. Turgidum (subunits CM2, CM3A, and CM16) and in T. tauschii (CM1, CM3D, and CM17) by a two-dimensional electrophoretic method that does not dissociate the inhibitors in the first dimension. Multiple tetrameric species, resulting from different combinations of the subunits contributed by the two ancestral species, are observed by the same procedure in T. aestivum. The three types of subunits were required for significant activity when the inhibitor of T. turgidum was reconstituted from the purified subunits, whereas, in the case of T. tauschii, binary mixtures involving subunit CM1 also had some activity. Additional combinations of the subunits present in these two species, which occur in the allohexaploid T. aestivum, were also reconstituted, and their inhibitory activities ranged from 144% to 33% the activity of the reconstituted inhibitor from T. tauschii. The activity of these inhibitors toward the ?-amylase (1,4-?-D-glucan glucanohydrolase, EC 220.127.116.11) of the insect Tenebrio molitor is much greater than that against the salivary enzyme. These observations, together with the previously established chromosomal locations of genes encoding CM proteins, fit a model of alloploid heterosis at the molecular level. Images PMID:16594035
Growth Factors Polymerized Within Fibrin Hydrogel Promote Amylase Production in Parotid Cells
McCall, Andrew D.; Nelson, Joel W.; Leigh, Noel J.; Duffey, Michael E.; Lei, Pedro; Andreadis, Stelios T.
Salivary gland cell differentiation has been a recurring challenge for researchers as primary salivary cells show a loss of phenotype in culture. Particularly, parotid cells show a marked decrease in amylase expression, the loss of tight junction organization and proper cell function. Previously, Matrigel has been used successfully as an extracellular matrix; however, it is not practical for in vivo applications as it is tumorigenic. An alternative method could rely on the use of fibrin hydrogel (FH), which has been used extensively in biomedical engineering applications ranging from cardiovascular tissue engineering to wound-healing experiments. Although several groups have examined the effects of a three-dimensional (3D) environment on salivary cell cultures, little is known about the effects of FH on salivary cell cultures. The current study developed a 3D cell culture model to support parotid gland cell differentiation using a combination of FH and growth factor-reduced Matrigel (GFR-MG). Furthermore, FH polymerized with a combination of EGF and IGF-1 induced formation of 3D spheroids capable of amylase expression and an agonist-induced increase in the intracellular Ca2+ concentration ([Ca2+]i) in salivary cells. These studies represent an initial step toward the construction of an artificial salivary gland to restore salivary gland dysfunction. This is necessary to reduce xerostomia in patients with compromised salivary function. PMID:23594102
Effects of tea polyphenols on the activities of ?-amylase, pepsin, trypsin and lipase
Qiang He; Yuanping Lv; Kai Yao
Tea polyphenols (TP) possess many beneficial properties, such as reducing the risk of cancer and heart diseases, and acting as natural antioxidants for the food industry. At the same time, tea polyphenols might inhibit digestive enzymes and reduce food digestibility. To explore this possible antinutritional property, the effects of tea polyphenols on the activity of four typical digestive enzymes were
Radionuclide salivary gland imaging
Mishkin, F.S.
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.
Adrenocortical activity in at-risk and normally developing adolescents: Individual differences in salivary cortisol basal levels, diurnal variation, and responses to social challenges
The purpose of this study was to examine adrenocortical activity (basal, diurnal variation, and responses to social stressors) in adolescents at risk for psychopathology. Salivary cortisol levels were examined in normally developing and at-risk youth with internalizing and externalizing symptoms ranging from subclinical to clinical levels. Adolescents showed expected patterns of diurnal variation, with high early morning cortisol levels and
Role of Streptococcus gordonii Amylase-Binding Protein A in Adhesion to Hydroxyapatite, Starch Metabolism, and Biofilm Formation
Rogers, Jeffrey D.; Palmer, Robert J.; Kolenbrander, Paul E.; Scannapieco, Frank A.
Interactions between bacteria and salivary components are thought to be important in the establishment and ecology of the oral microflora. ?-Amylase, the predominant salivary enzyme in humans, binds to Streptococcus gordonii, a primary colonizer of the tooth. Previous studies have implicated this interaction in adhesion of the bacteria to salivary pellicles, catabolism of dietary starches, and biofilm formation. Amylase binding is mediated at least in part by the amylase-binding protein A (AbpA). To study the function of this protein, an erythromycin resistance determinant [erm(AM)] was inserted within the abpA gene of S. gordonii strains Challis and FAS4 by allelic exchange, resulting in abpA mutant strains Challis-E1 and FAS4-E1. Comparison of the wild-type and mutant strains did not reveal any significant differences in colony morphology, biochemical metabolic profiles, growth in complex or defined media, surface hydrophobicity, or coaggregation properties. Scatchard analysis of adhesion isotherms demonstrated that the wild-type strains adhered better to human parotid-saliva- and amylase-coated hydroxyapatite than did the AbpA mutants. In contrast, the mutant strains bound to whole-saliva-coated hydroxyapatite to a greater extent than did the wild-type strains. While the wild-type strains preincubated with purified salivary amylase grew well in defined medium with potato starch as the sole carbohydrate source, the AbpA mutants did not grow under the same conditions even after preincubation with amylase. In addition, the wild-type strain produced large microcolonies in a flow cell biofilm model, while the abpA mutant strains grew much more poorly and produced relatively small microcolonies. Taken together, these results suggest that AbpA of S. gordonii functions as an adhesin to amylase-coated hydroxyapatite, in salivary-amylase-mediated catabolism of dietary starches and in human saliva-supported biofilm formation by S. gordonii. PMID:11598080
Role of Streptococcus gordonii amylase-binding protein A in adhesion to hydroxyapatite, starch metabolism, and biofilm formation.
Rogers, J D; Palmer, R J; Kolenbrander, P E; Scannapieco, F A
Interactions between bacteria and salivary components are thought to be important in the establishment and ecology of the oral microflora. alpha-Amylase, the predominant salivary enzyme in humans, binds to Streptococcus gordonii, a primary colonizer of the tooth. Previous studies have implicated this interaction in adhesion of the bacteria to salivary pellicles, catabolism of dietary starches, and biofilm formation. Amylase binding is mediated at least in part by the amylase-binding protein A (AbpA). To study the function of this protein, an erythromycin resistance determinant [erm(AM)] was inserted within the abpA gene of S. gordonii strains Challis and FAS4 by allelic exchange, resulting in abpA mutant strains Challis-E1 and FAS4-E1. Comparison of the wild-type and mutant strains did not reveal any significant differences in colony morphology, biochemical metabolic profiles, growth in complex or defined media, surface hydrophobicity, or coaggregation properties. Scatchard analysis of adhesion isotherms demonstrated that the wild-type strains adhered better to human parotid-saliva- and amylase-coated hydroxyapatite than did the AbpA mutants. In contrast, the mutant strains bound to whole-saliva-coated hydroxyapatite to a greater extent than did the wild-type strains. While the wild-type strains preincubated with purified salivary amylase grew well in defined medium with potato starch as the sole carbohydrate source, the AbpA mutants did not grow under the same conditions even after preincubation with amylase. In addition, the wild-type strain produced large microcolonies in a flow cell biofilm model, while the abpA mutant strains grew much more poorly and produced relatively small microcolonies. Taken together, these results suggest that AbpA of S. gordonii functions as an adhesin to amylase-coated hydroxyapatite, in salivary-amylase-mediated catabolism of dietary starches and in human saliva-supported biofilm formation by S. gordonii. PMID:11598080
Wong, D T W
Saliva is a noninvasive and accessible biofluid that permits early detection of oral and systemic diseases. Recent scientific and technologic advances have uncovered specific salivary biomarkers for a number of clinical conditions, including cancers, autoimmune diseases, and cardiovascular disorders. The availability of highly sensitive and high-throughput assays such as microarray, mass spectrometry, reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) and nano-scale sensors that can measure proteins and nucleic acids are poising saliva as an emerging biofluid for translational and clinical applications. This paper will discuss development of salivary biomarkers for the detection of oral and systemic diseases and the translational application of these markers for clinical applications. PMID:22788724
Identification and characterization of amylase binding protein C (AbpC) from Streptococcus mitis NS51
Vorrasi, John; Chaudhuri, Biswendu; Haase, Elaine M.; Scannapieco, Frank A.
SUMMARY A substantial proportion of the streptococcal species found in dental plaque biofilms are able to interact with the abundant salivary enzyme ?-amylase. These streptococci produce proteins that specifically bind amylase. An important plaque species, Streptococcus mitis, secretes a 36-kDa amylase binding protein into the extracellular milieu. Proteins precipitated from S. mitis NS51 cell culture supernatant by the addition of purified salivary amylase were separated by SDS-PAGE, transferred to a membrane, and a prominent 36-kDa band was cut from the membrane and sequencedto yield N-terminal amino acid sequence DSQAQYSNGV. Search of the S. mitis genome sequence database revealed a single open reading frame containing this sequence, and the gene was amplified from S. mitis genomic DNA polymerase chain reaction. The coding region of this ORF, designated amylase-binding protein C (AbpC), was cloned into an Escherichia coli expression vectorand the recombinant AbpC protein (rAbpC) was purified from the soluble fraction of E. coli cell lysate. Purified AbpC was found to interact with immobilized amylase, thus confirming AbpC as a new streptococcal amylase-binding protein. PMID:20331802
Identification and characterization of amylase-binding protein C from Streptococcus mitis NS51.
Vorrasi, J; Chaudhuri, B; Haase, E M; Scannapieco, F A
A substantial proportion of the streptococcal species found in dental plaque biofilms are able to interact with the abundant salivary enzyme alpha-amylase. These streptococci produce proteins that specifically bind amylase. An important plaque species, Streptococcus mitis, secretes a 36-kDa amylase-binding protein into the extracellular milieu. Proteins precipitated from S. mitis NS51 cell culture supernatant by the addition of purified salivary amylase were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to a membrane, and a prominent 36-kDa band was cut from the membrane and sequenced to yield the N-terminal amino acid sequence DSQAQYSNGV. Searching the S. mitis genome sequence database revealed a single open reading frame containing this sequence, and the gene was amplified by the S. mitis genomic DNA polymerase chain reaction. The coding region of this open reading frame, designated amylase-binding protein C (AbpC), was cloned into an Escherichia coli expression vector and the recombinant AbpC (rAbpC) was purified from the soluble fraction of the E. coli cell lysate. Purified AbpC was found to interact with immobilized amylase, confirming AbpC as a new streptococcal amylase-binding protein. PMID:20331802
Effect of introducing a disulphide bond between the A and C domains on the activity and stability of Saccharomycopsis fibuligera R64 ?-amylase.
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
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 0.45 mM, nearly two times lower than that of Sfamy01 at 0.77 mM. 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
Salivary gland dysfunction following radioactive iodine therapy
Wiesenfeld, D.; Webster, G.; Cameron, F.; Ferguson, M.M.; MacFadyen, E.E.; MacFarlane, T.W.
Radioactive iodine is used extensively for the treatment of thyrotoxicosis and thyroid carcinoma. Iodine is actively taken up by the salivary glands and, following its use, salivary dysfunction may result as a consequence of radiation damage. The literature is reviewed and a case is reported in which a patient presented with a significant increase in caries rate attributed to salivary dysfunction following radioactive iodine therapy for a thyroid carcinoma.
Microcalorimetric study of the enzymatic hydrolysis of starch: An ?-amylase catalyzed reaction
G. Salieri; G. Vinci; M. L. Antonelli
Microcalorimetric studies of enzyme activities have been made on the hydrolysis of starch catalyzed by ?-amylase. The best conditions for the hydrolysis of starch in the presence of ?-amylase have been pointed out and the ?-amylase activity was determined by isothermal batch microcalorimetry. The heat changes involved during the hydrolytic reaction can be related to the starch concentrations in a
Effects of temperature on the endogenous activity and synaptic interactions of the salivary burster neurones in the terrestrial slug Limax maximus.
Prior, D J; Grega, D S
(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
Effect of salivary gland adenocarcinoma cell-derived alpha-N-acetylgalactosaminidase on the bioactivity of macrophage activating factor.
Matsuura, Takashi; Uematsu, Takashi; Yamaoka, Minoru; Furusawa, Kiyofumi
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
Production of Fungal Amylases Using Cheap, Readily Available Agriresidues, for Potential Application in Textile Industry
Singh, Sanamdeep; Bali, Vrinda; Mangla, Jyoti
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
?-Amylases of the coffee berry borer ( Hypothenemus hampei) and their inhibition by two plant amylase inhibitors
Arnubio Valencia; Alex E Bustillo; Gustavo E Ossa; Maarten J Chrispeels
The adult coffee berry borer (Hypothenemus hampei Ferrari [Coleoptera: Scolytidae]), a major insect pest of coffee, has two major digestive ?-amylases that can be separated by isoelectric focusing. The ?-amylase activity has a broad pH optimum between 4.0 and 7.0. Using pH indicators, the pH of the midgut was determined to be between 4.5 and 5.2. At pH5.0, the coffee
Flaxseed (Linum usitatissimum L.) extract as well as (+)-secoisolariciresinol diglucoside and its mammalian derivatives are potent inhibitors of ?-amylase activity.
Hano, Christophe; Renouard, Sullivan; Molinié, Roland; Corbin, Cyrielle; Barakzoy, Esmatullah; Doussot, Joël; Lamblin, Frédéric; Lainé, Eric
Type 2 diabetes mellitus (T2DM) is one of the common global diseases. Flaxseed is by far the richest source of the dietary lignans (i.e., secoisolariciresinol diglucoside) which have been shown to delay the development of T2DM in animal models. Herein, we propose the first evidences for a mechanism of action involving the inhibition of the pancreatic ?-amylase (EC 188.8.131.52) by flaxseed-derived lignans that could therefore constitute a promising nutraceutical for the prevention and the treatment of T2DM. PMID:23583514
Salivary contribution to exhaled nitric oxide
W. Zetterquist; C. Pedroletti; J. O. n. Lundberg; K. Alving
Dietary and metabolic nitrate is distributed from the blood to the saliva by active uptake in the salivary glands, and is reduced to nitrite in the oral cavity by the action of certain bacteria. Since it has been reported that nitric oxide may be formed nonenzymatically from nitrite this study aimed to determine whether salivary nitrite could influence measurements of
Relationship of long-term highly active antiretroviral therapy on salivary flow rate and CD4 Count among HIV-infected patients
Kumar, J Vijay; Baghirath, P Venkat; Naishadham, P Parameswar; Suneetha, Sujai; Suneetha, Lavanya; Sreedevi, P
Objectives: To determine if long-term highly active antiretroviral therapy (HAART) therapy alters salivary flow rate and also to compare its relation of CD4 count with unstimulated and stimulated whole saliva. Materials and Methods: A cross-sectional study was performed on 150 individuals divided into three groups. Group I (50 human immunodeficiency virus (HIV) seropositive patients, but not on HAART therapy), Group II (50 HIV-infected subjects and on HAART for less than 3 years called short-term HAART), Group III (50 HIV-infected subjects and on HAART for more than or equal to 3 years called long-term HAART). Spitting method proposed by Navazesh and Kumar was used for the measurement of unstimulated and stimulated salivary flow rate. Chi-square test and analysis of variance (ANOVA) were used for statistical analysis. Results: The mean CD4 count was 424.78 ± 187.03, 497.82 ± 206.11 and 537.6 ± 264.00 in the respective groups. Majority of the patients in all the groups had a CD4 count between 401 and 600. Both unstimulated and stimulated whole salivary (UWS and SWS) flow rates in Group I was found to be significantly higher than in Group II (P < 0.05). Unstimulated salivary flow rate between Group II and III subjects were also found to be statistically significant (P < 0.05). ANOVA performed between CD4 count and unstimulated and stimulated whole saliva in each group demonstrated a statistically significant relationship in Group II (P < 0.05). There were no significant results found between CD4 count and stimulated whole saliva in each groups. Conclusion: The reduction in CD4 cell counts were significantly associated with salivary flow rates of HIV-infected individuals who are on long-term HAART.
Suppression of white light generation (supercontinuum) in biological media: a pilot study using human salivary proteins
NASA Astrophysics Data System (ADS)
Santhosh, C.; Dharmadhikari, A. K.; Alti, K.; Dharmadhikari, J. A.; Mathur, D.
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.
Primary structure of the active tryptic fragments of human and monkey salivary anionic proline-rich proteins.
Schlesinger, D H; Hay, D I
The primary structures of the four human salivary anionic proline-rich proteins and an analogous protein from the saliva of a monkey (Macaca arctoides) have been further investigated. These proteins possess the unusual property of inhibiting crystal growth of calcium phosphate salts, and it has been proposed that they play an important role in the mouth, by preventing precipitation of calcium phosphate salts from the supersaturated saliva. The tryptic fragments responsible for this activity have been isolated from all five proteins and their complete amino acid sequences determined and compared. These active fragments have been unequivocally identified as the amino-terminal segment in all five proteins. The structures of the four human fragments are identical except for the presence of Asn at residue 4 in PRP-1 and -3 instead of Asp found in PRP-2 and -4. Comparison of the 30 residue human fragments with the monkey fragment shows 18 residues to be identical in these peptides, providing that residue 1 of the monkey fragment is aligned with residue 3 of the human proteins. Theses studies constitute the next step in determining the mechanism of action of these unusual proteins, and in determining their minimum chain length required for inhibitory activity. PMID:7228490
The effectiveness of the Uchida-Kraepelin test for psychological stress: an analysis of plasma and salivary stress substances
Sugimoto, Koreaki; Kanai, Aya; Shoji, Noriaki
Background The hypothalamic-pituitary-adrenocortical (HPA) axis and sympathetic adrenomedullary (SAM) system are the major stress-response pathways. Plasma adrenocorticotropic hormone (ACTH) represents HPA axis activity, while plasma catecholamines are used as markers of the SAM system. Salivary alpha amylase (AA), chromogranin A (CgA), and immunoglobulin A (IgA) are candidate markers of stress activation, although their role has not been established. The Uchida-Kraepelin (U-K) test is a questionnaire that requires intense concentration and effort, and has been used as a tool to induce mental stress. However, it is not clear whether or not the test is effective as a psychological/mental stressor. Methods In this study, normal young women took the U-K test and serial measurements of plasma ACTH and catecholamines (dopamine, noradrenaline, and adrenaline) (n = 10), as well as salivary AA, CgA, and IgA (n = 16) before, during and after the test. Results We found no changes in any of these parameters at any time point during or after the U-K test. Conclusion Our findings indicate that the U-K test is not a suitable for measuring the psychological/mental stress of young women because the plasma data showed that it did not affect the HPA axis and SAM system. The U-K test should be employed carefully as a psychological/mental stressor due to insufficient scientific evidence of its effectiveness. In addition, salivary AA, CgA, and IgA should not simply be compared with previous reports, because the mechanism of secretion and normal range of each salivary parameter remain unknown. Salivary AA, CgA, and IgA may not be suitable candidate markers of psychological/mental stress. PMID:19341484
The Rapalogue, CCI-779, Improves Salivary Gland Function following Radiation
Morgan-Bathke, Maria; Harris, Zoey I.; Arnett, Deborah G.; Klein, Rob R.; Burd, Randy; Ann, David K.; Limesand, Kirsten H.
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
Investigating the role of perceived stress on bacterial flora activity and salivary cortisol secretion: A possible mechanism underlying susceptibility to illness
Simon R. Knowles; Elizabeth A. Nelson; Enzo A. Palombo
This study examined the impact of academic stress on salivary cortisol concentrations and lactic acid bacteria activity. Whole, unstimulated saliva samples and faecal samples were collected from 23 healthy undergraduate students (23.0±6.8 years; range 18–44) over two 1-week periods: during the beginning of semester (low-stress baseline condition) and during the first week of exams (high-stress condition). Students also completed a
HPLC-DAD Analysis and In-Vitro Property of Polyphenols Extracts from (Solanum Aethiopium) Fruits on ? -Amylase, ? -Glucosidase and Angiotensin - 1- Converting Enzyme Activities
Nwanna, E. E; Ibukun, E. O; Oboh, G.; Ademosun, A. O.; Boligon, A. A.; Athayde, M.
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
Investigation of insecticidal activity of rye ?-amylase inhibitor gene expressed in transgenic tobacco ( Nicotiana tabacum) toward cotton boll weevil ( Anthonomus grandis)
Simoni Campos Dias; Maria Cristina Mattar da Silva; Fábíola R. Teixeira; Edson Luis Zangrando Figueira; Osmundo Brilhante de Oliveira-Neto; Loaiane Alves de Lima; Octávio Luiz Franco; Maria Fátima Grossi-de-Sa
Innumerable proteinaceous ?-amylase inhibitors have been isolated and identified from different plant species. Among them, an ?-amylase inhibitor gene with bioinsecticidal potential toward Anthonomus grandis (cotton boll weevil) was previously identified in rye seeds (Secale cereale). This cereal inhibitor was expressed in tobacco plants (Nicotiana tabacum) under control of phytohemaglutinin promoter by using Agrobacterium tumefasciens – mediated transformation. Presence of
Microbial ?-amylases: a biotechnological perspective
Rani Gupta; Paresh Gigras; Harapriya Mohapatra; Vineet Kumar Goswami; Bhavna Chauhan
Amylases are one of the most important and oldest industrial enzymes. These comprise hydrolases, which hydrolyse starch molecules to fine diverse products as dextrins, and progressively smaller polymers composed of glucose units. Large arrays of amylases are involved in the complete breakdown of starch. However, ?-amylases which are the most in demand hydrolyse ?-1,4 glycosidic bond in the interior of
Protein engineering of bacterial ?-amylases
Jens Erik Nielsen; Torben V. Borchert
?-Amylases constitute a very diverse family of glycosyl hydrolases that cleave ?1?4 linkages in amylose and related polymers. Recent structural and mutagenic studies of archeael, mammalian and bacterial ?-amylases have resulted in a wealth of information on the catalytic mechanism and on the structural features of this enzyme class. Because of their high thermo-stability, the Bacillus ?-amylases have found widespread
Demonstration of amylase from the zoophytophagous anthocorid Orius insidiosus.
Zeng, F; Cohen, A C
To better understand facultative phytophagy in the zoophytophagous anthocorid, Orius insidiosus, tests for amylase were conducted and the enzyme was partially purified. Three activity bands were detected with polyacrylamide-starch gel electrophoretic analysis of amylase in O. insidiosus. The major amylase was found to have a mean isoelectric point (pI) of 4.53. The presence of amylase indicates the ability of O. insidiosus to use starch, a nutrient, derived from plants, either by direct ingestion or by ingestion of plant material from the digestive system of their prey. The presence of amylase suggests that these predators are more committed to plant feeding than other species of predators that lack this enzyme. PMID:10897095
Mass Spectrometric Analysis of Whole Secretome and Amylase-precipitated Secretome Proteins from Streptococcus gordonii
Maddi, A; Haase, EM; Scannapieco, FA
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
Lufaxin, a Novel Factor Xa Inhibitor from the Salivary Gland of the sand fly Lutzomyia longipalpis, Blocks PAR2 Activation and Inhibits Inflammation and Thrombosis in Vivo
Collin, Nicolas; Assumpção, Teresa C. F.; Mizurini, Daniella M.; Gilmore, Dana; Dutra-Oliveira, Angélica; Kotsyfakis, Michalis; Sá-Nunes, Anderson; Teixeira, Clarissa; Ribeiro, José M. C.; Monteiro, Robson Q.; Valenzuela, Jesus G.; Francischetti, Ivo M. B.
Objective Blood-sucking arthropods salivary glands (SGs) contain a remarkable diversity of antihemostatics. The aim of this study was to identify the unique salivary anticoagulant of the sand fly Lutzomyia longipalpis, which remained elusive for decades. Methods and Results Several L. longipalpis salivary proteins were expressed in HEK293 cells and screened for inhibition of blood coagulation. A novel 32.4-kDa molecule, named Lufaxin, was identified as a slow, tight, non-competitive, and reversible inhibitor of Factor Xa (FXa). Notably, Lufaxin primary sequence does not share similarity to any physiological or salivary inhibitors of coagulation reported to date. Lufaxin is specific for FXa and does not interact with FX, DEGR- FXa, or 15 other enzymes. In addition, Lufaxin blocks prothrombinase and increases both PT and aPTT. Surface plasmon resonance experiments revealed that FXa binds Lufaxin with a KD ~3 nM, and isothermal titration calorimetry determined a stoichiometry of 1:1. Lufaxin also prevents PAR2 activation by FXa in the MDA-MB-231 cell line and abrogates edema formation triggered by injection of FXa in the paw of mice. Moreover, Lufaxin prevents FeCl3-induced carotid artery thrombus formation and prolongs aPTT ex vivo, implying that it works as an anticoagulant in vivo. Finally, SG of sandflies was found to inhibit FXa and to interact with the enzyme. Conclusion Lufaxin belongs to a novel family of slow-tight FXa inhibitors, which display antithrombotic and antiinflamatory activities. It is a useful tool to understand FXa structural features and its role in pro-hemostatic and pro-inflammatory events. PMID:22796577
GA Enhanced a-Amylase Synthesis in Halved Grains of Barley (Hordeum vulgare): A Simple Laboratory Demonstration
ERIC Educational Resources Information Center
Freeland, P. W.
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)
Alpha-amylase reactivity in relation to psychopathic traits in adults.
Glenn, Andrea L; Remmel, Rheanna J; Raine, Adrian; Schug, Robert A; Gao, Yu; Granger, Douglas A
Recent investigations of the psychobiology of stress in antisocial youth have benefited from a multi-system measurement model. The inclusion of salivary alpha-amylase (sAA), a surrogate marker of autonomic/sympathetic nervous system (ANS) activity, in addition to salivary cortisol, a biomarker of the hypothalamic-pituitary-adrenal (HPA) axis functioning, has helped define a more complete picture of individual differences and potential dysfunction in the stress response system of these individuals. To the authors' knowledge, no studies have examined sAA in relation to antisocial behavior in adults or in relation to psychopathic traits specifically. In the present study, we examined sAA, in addition to salivary cortisol, in a relatively large sample (n=158) of adult males (M age=36.81, range=22-67 years; 44% African-American, 34% Caucasian, 16% Hispanic) recruited from temporary employment agencies with varying levels of psychopathic traits. Males scoring highest in psychopathy were found to have attenuated sAA reactivity to social stress compared to those scoring lower in psychopathy. No differential relationships with the different factors of psychopathy were observed. In contrast to studies of antisocial youth, there were no interactions between sAA and cortisol levels in relation to psychopathy, but there was a significant interaction between pre-stressor levels of sAA and cortisol. Findings reveal potential regulatory deficits in the fast-acting, 'fight or flight', component of the stress response in adult males with psychopathic traits, as well as abnormalities in how this system may interact with the HPA axis. PMID:25662339
High-resolution ?-amylase assay combined with high-performance liquid chromatography-solid-phase extraction-nuclear magnetic resonance spectroscopy for expedited identification of ?-amylase inhibitors: proof of concept and ?-amylase inhibitor in cinnamon.
Okutan, Leyla; Kongstad, Kenneth T; Jäger, Anna K; Staerk, Dan
Type 2 diabetes affects millions of people worldwide, and new improved drugs or functional foods containing selective ?-amylase inhibitors are needed for improved management of blood glucose. In this article the development of a microplate-based high-resolution ?-amylase inhibition assay with direct photometric measurement of ?-amylase activity is described. The inhibition assay is based on porcine pancreatic ?-amylase with 2-chloro-4-nitrophenyl-?-D-maltotriose as substrate, which this gives a stable, sensitive, and cheap inhibition assay as requested for high-resolution purposes. In combination with HPLC-HRMS-SPE-NMR, this provides an analytical platform that allows simultaneous chemical and biological profiling of ?-amylase inhibitors in plant extracts. Proof-of-concept with an artificial mixture of six compounds-of which three are known ?-amylase inhibitors-showed that the high-resolution ?-amylase inhibition profiles allowed detection of sub-microgram amounts of the ?-amylase inhibitors. Furthermore, the high-resolution ?-amylase inhibition assay/HPLC-HRMS-SPE-NMR platform allowed identification of cinnamaldehyde as the ?-amylase inhibitor in cinnamon (Cinnamomum verum Presl.). PMID:25368916
Structural features of salivary function.
Lamkin, M S; Oppenheim, F G
Saliva plays an important role in the maintenance of oral health by exhibiting multiple host defense functions. These include homeostatic processes, lubrication, antimicrobial activity, and the control of demineralization/remineralization of teeth. Biochemical studies of saliva and salivary secretions established that specific salivary proteins are responsible for these defense functions. Because some of these salivary proteins have been characterized extensively, including their primary structures, it has become feasible to explore their structure/function relationships. Acidic proline-rich proteins (PRPs), for example, exhibit high affinity to hydroxyapatite, inhibit crystal growth of calcium phosphate salts from solutions supersaturated with respect to hydroxyapatite, bind calcium ions, and interact with several oral bacteria on adsorption to hydroxyapatite. Statherins, histatins, and cystatins also exhibit affinities to mineral surfaces, inhibit calcium phosphate precipitation, and play a role in maintaining the integrity of teeth. Furthermore, histatins exhibit both antibacterial and antifungal activities. Approaches to identifying the functional domains of these salivary proteins include functional assays of enzymatically digested proteins and peptides, synthetic peptides and peptide analogues, and chemically modified proteins as well as biophysical studies of native proteins or peptides. Such studies have demonstrated that the fungicidal activities of histatins reside in the middle portion of the polypeptide chain, whereas the hydroxyapatite binding domains of PRPs and statherin reside in the phosphorylated amino-terminal regions. Identification of functional domains is vital in understanding the mechanisms of action and this information can be exploited in the development of therapeutic agents. PMID:8373982
The Enzyme-Like Domain of Arabidopsis Nuclear ?-Amylases Is Critical for DNA Sequence Recognition and Transcriptional Activation[C][W][OPEN
Soyk, Sebastian; Šimková, Klára; Zürcher, Evelyne; Luginbühl, Leonie; Brand, Luise H.; Vaughan, Cara K.; Wanke, Dierk; Zeeman, Samuel C.
Plant BZR1-BAM transcription factors contain a ?-amylase (BAM)–like domain, characteristic of proteins involved in starch breakdown. The enzyme-derived domains appear to be noncatalytic, but they determine the function of the two Arabidopsis thaliana BZR1-BAM isoforms (BAM7 and BAM8) during transcriptional initiation. Removal or swapping of the BAM domains demonstrates that the BAM7 BAM domain restricts DNA binding and transcriptional activation, while the BAM8 BAM domain allows both activities. Furthermore, we demonstrate that BAM7 and BAM8 interact on the protein level and cooperate during transcriptional regulation. Site-directed mutagenesis of residues in the BAM domain of BAM8 shows that its function as a transcriptional activator is independent of catalysis but requires an intact substrate binding site, suggesting it may bind a ligand. Microarray experiments with plants overexpressing truncated versions lacking the BAM domain indicate that the pseudo-enzymatic domain increases selectivity for the preferred cis-regulatory element BBRE (BZR1-BAM Responsive Element). Side specificity toward the G-box may allow crosstalk to other signaling networks. This work highlights the importance of the enzyme-derived domain of BZR1-BAMs, supporting their potential role as metabolic sensors. PMID:24748042
Candida albicans Flu1-Mediated Efflux of Salivary Histatin 5 Reduces Its Cytosolic Concentration and Fungicidal Activity
Li, Rui; Kumar, Rohitashw; Tati, Swetha; Puri, Sumant
Histatin 5 (Hst 5) is a salivary human antimicrobial peptide that is toxic to the opportunistic yeast Candida albicans. Fungicidal activity of Hst 5 requires intracellular translocation and accumulation to a threshold concentration for it to disrupt cellular processes. Previously, we observed that total cytosolic levels of Hst 5 were gradually reduced from intact cells, suggesting that C. albicans possesses a transport mechanism for efflux of Hst 5. Since we identified C. albicans polyamine transporters responsible for Hst 5 uptake, we hypothesized that one or more polyamine efflux transporters may be involved in the efflux of Hst 5. C. albicans FLU1 and TPO2 were found to be the closest homologs of Saccharomyces cerevisiae TPO1, which encodes a major spermidine efflux transporter, indicating that the products of these two genes may be involved in efflux of Hst 5. We found that flu1?/? cells, but not tpo2?/? cells, had significant reductions in their rates of Hst 5 efflux and had significantly higher cytoplasmic Hst 5 and Hst 5 susceptibilities than did the wild type. We also found that flu1?/? cells had reduced biofilm formation compared to wild-type cells in the presence of Hst 5. Transcriptional levels of FLU1 were not altered over the course of treatment with Hst 5; therefore, Hst 5 is not likely to induce FLU1 gene overexpression as a potential mechanism of resistance. Thus, Flu1, but not Tpo2, mediates efflux of Hst 5 and is responsible for reduction of its toxicity in C. albicans. PMID:23380720
What interactions drive the salivary mucosal pellicle formation?
Gibbins, Hannah L.; Yakubov, Gleb E.; Proctor, Gordon B.; Wilson, Stephen; Carpenter, Guy H.
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
Salivary proteins of plant-feeding hemipteroids - implication in phytophagy.
Sharma, A; Khan, A N; Subrahmanyam, S; Raman, A; Taylor, G S; Fletcher, M J
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
Rle de la muqueuse duodnale dans l'adaptation de l'?-amylase pancratique au rgime alimentaire chez le Porc
Paris-Sud XI, Université de
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
Biochemical Characterization and Mass Spectrometric Disulfide Bond Mapping of Periplasmic -Amylase MalS of Escherichia coli*
Rippe, Karsten
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
Potential role of lysozyme in bactericidal activity of in vitro-acquired salivary pellicle against Streptococcus faecium 9790.
Germaine, G R; Tellefson, L M
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
Inhibition of calcium phosphate precipitation by human salivary statherin: structure-activity relationships.
Schwartz, S S; Hay, D I; Schluckebier, S K
Previous studies of human statherin showed the active region for inhibition of secondary calcium phosphate precipitation (crystal growth) to reside in the highly charged amino-terminal one-third of this molecule, and the neutral tyrosine-, glutamine- and proline-rich carboxy-terminal two-thirds of the molecule is required for maximal inhibition of primary (spontaneous) precipitation. The purpose of the present study was to define more clearly the activities of these different molecular segments of statherin with respect to the two kinds of inhibitory activities. Peptides from statherin were prepared by specific proteolysis using trypsin, endoproteinase Arg-C, and activated factor X to produce the amino-terminal hexa-, nona- and decapeptides, respectively, and carboxypeptidase-A was used to obtain a peptide extending from residue 1 to about residues 32-37. The peptides were purified by anion exchange and gel filtration chromatography, and characterized and quantified by amino-acid analysis. Serially diluted samples of statherin and derived peptides were assayed to determine the concentrations, giving a standard 50% inhibition of precipitation (C50%) in assay systems designed for this purpose using polyaspartate as a standard. Results are expressed as (C50% statherin)/(C50% peptide). For inhibition of primary precipitation, these values were peptide(1-6), 0.20; peptide(1-9), 0.15; peptide(1-31/35), 0.24. For inhibition of secondary precipitation, the values were peptide(1-6), 3.8; peptide(1-9), 2.8; peptide(1-10), 1.9; peptide(1-32/37), 1.5. These quantitative findings show that maximum inhibition of primary precipitation by statherin requires the entire molecule. Thus, removal of a relatively small segment of its carboxy-terminal region results in a substantial reduction in inhibitory activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1525706
Use of a replica-plate assay for the rapid assessment of salivary protein-bacteria interactions.
Tseng, C C; Scannapieco, F A; Levine, M J
A replica-plate assay was used to screen for the interaction of salivary molecules with dental plaque bacteria. Bacterial colonies cultured from supragingival plaque on sheep-blood (SB) agar were replica-plated onto nitrocellulose membranes overlaying SB or mitis-salivarius agar. Membranes with attached colonies were removed and incubated with 125I-amylase or 125I-proline-rich glycoprotein (PRG). Positive interactions were detected by autoradiography. Only strains of Streptococcus gordonii and Actinomyces viscosus bound amylase, and strains of A. viscosus bound PRG. The results suggest that amylase and PRG bind to selected species of aerobic dental plaque bacteria. PMID:1382259
Activation of histamine H4 receptor inhibits TNF?/IMD-0354-induced apoptosis in human salivary NS-SV-AC cells.
Stegajev, Vasili; Kouri, Vesa-Petteri; Salem, Abdelhakim; Rozov, Stanislav; Stark, Holger; Nordström, Dan C E; Konttinen, Yrjö T
Apoptosis is involved in the pathogenesis of Sjögren's syndrome (SS), an autoimmune disease affecting exocrine glands. Our recent studies revealed diminished histamine H4 receptor (H?R) expression and impaired histamine transport in the salivary gland epithelial cells in SS. The aim was now to test if nanomolar histamine and high-affinity H?R signaling affect apoptosis of human salivary gland epithelial cell. Simian virus 40-immortalized acinar NS-SV-AC cells were cultured in serum-free keratinocyte medium ± histamine H?R agonist HST-10. Expression and internalization of H?R were studied by immunofluorescence staining ± clathrin inhibitor methyl-?-cyclodextrin (M?CD). Apoptosis induced using tumor necrosis factor-? with nuclear factor-?B inhibitor IMD-0354 was studied using phase contrast microscopy, Western blot, flow cytometry and polymerase chain reaction (qRT-PCR). HST-10-stimulated H?R internalization was inhibited by M?CD. Western blotting revealed diminished phosphorylated c-Jun N-terminal kinase JNK, but unchanged levels of phosphorylated extracellular signal regulated kinase pERK1/2 in H?R-stimulated samples compared to controls. qRT-PCR showed up-regulated expression of anti-apoptotic B cell lymphoma-extra large/Bcl-xL mRNAs and proteins, whereas pro-apoptotic Bcl-2-associated X protein/BAX remained unchanged in H4R-stimulated samples. H?R stimulation diminished cleavage of PARP and flow cytometry showed significant dose-dependent inhibitory effect of H?R stimulation on apoptosis. As far as we know this is the first study showing inhibitory effect of H?R activation on apoptosis of human salivary gland cells. Diminished H?R-mediated activation may contribute to loss of immune tolerance in autoimmune diseases and in SS in particular. PMID:25239604
Stabilization of ?-amylase by using anionic surfactant during the immobilization process
NASA Astrophysics Data System (ADS)
El-Batal, A. I.; Atia, K. S.; Eid, M.
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.
The impact of a 24-h ultra-marathon on salivary antimicrobial protein responses.
Gill, S K; Teixeira, A M; Rosado, F; Hankey, J; Wright, A; Marczak, S; Murray, A; Costa, R J S
Depressed oral respiratory mucosal immunity and increased incidence of upper respiratory symptoms are commonly reported after bouts of prolonged exercise. The current study observed the impact of a 24-h continuous overnight ultra-marathon competition (distance range: 122-208?km; ambient temperature range: 0-20 °C) on salivary antimicrobial protein responses and incidence of upper respiratory symptoms. Body mass, unstimulated saliva and venous blood samples were taken from ultra-endurance runners (n=25) and controls (n=17), before and immediately after competition. Upper respiratory symptoms were assessed during and until 4-weeks after event completion. Samples were analyzed for salivary IgA, lysozyme, ?-amylase and cortisol in addition to plasma osmolality. Decreased saliva flow rate (p<0.001), salivary IgA (p<0.001) and lysozyme (p=0.015) secretion rates, and increased salivary ?-amylase secretion rate (p<0.001) and cortisol responses (p<0.001) were observed post-competition in runners, with no changes being observed in controls. No incidences of upper respiratory symptoms were reported by participants. A 24-h continuous overnight ultra-marathon resulted in the depression of some salivary antimicrobial protein responses, but no incidences of upper respiratory symptoms were evident during or following competition. Salivary antimicrobial protein synergism, effective management of non-infectious episodes, maintaining euhydration, and (or) favourable environmental influences could have accounted for the low prevalence of upper respiratory symptoms. PMID:24886918
Implantable Three-Dimensional Salivary Spheroid Assemblies Demonstrate Fluid and Protein Secretory Responses to Neurotransmitters
Pradhan-Bhatt, Swati; Harrington, Daniel A.; Duncan, Randall L.; Jia, Xinqiao; Witt, Robert L.
Radiation treatment in patients with head and neck tumors commonly results in hyposalivation and xerostomia due to the loss of fluid-secreting salivary acinar cells. Patients develop susceptibility to oral infections, dental caries, impaired speech and swallowing, reducing the quality of life. Clinical management is largely unsatisfactory. The development of a tissue-engineered, implantable salivary gland will greatly benefit patients suffering from xerostomia. This report compares the ability of a 2.5-dimensional (2.5D) and a three-dimensional (3D) hyaluronic acid (HA)-based culture system to support functional salivary units capable of producing fluid and phenotypic proteins. Parotid cells seeded on 2.5D, as well as those encapsulated in 3D HA hydrogels, self-assembled into acini-like structures and expressed functional neurotransmitter receptors. Structures in 3D hydrogels merged to form organized 50??m spheroids that could be maintained in culture for over 100 days and merged to form structures over 500??m in size. Treatment of acini-like structures with the ?-adrenergic agonists norepinephrine or isoproterenol increased granule production and ?-amylase staining in treated structures, demonstrating regain of protein secretion. Upon treatment with the M3 muscarinic agonist acetylcholine, acini-like structures activated the fluid production pathway by increasing intracellular calcium levels. The increase in intracellular calcium seen in structures in the 3D hydrogel culture system was more robust and prolonged than that in 2.5D. To compare the long-term survival and retention of acini-like structures in vivo, cell-seeded 2.5D and 3D hydrogels were implanted into an athymic rat model. Cells in 2.5D failed to maintain organized acini-like structures and dispersed in the surrounding tissue. Encapsulated cells in 3D retained their spheroid structure and structural integrity, along with the salivary biomarkers and maintained viability for over 3 weeks in vivo. This report identifies a novel hydrogel culture system capable of creating and maintaining functional 3D salivary spheroid structures for long periods in vitro that regain both fluid and protein secreting functions and are suitable for tissue restoration. PMID:23442148
Anopheles salivary gland proteomes from major malaria vectors
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
An efficient purification process for sweet potato beta-amylase by affinity precipitation with alginate.
Teotia, S; Khare, S K.; Gupta, M N.
beta-amylases are used in production of maltose syrup. It is shown that sweet potato beta-amylase can be purified by affinity precipitation with alginate with 80% activity yield and 44 fold purification. SDS-PAGE of the purified protein showed a single band and a subunit weight of 50 kDa. Preliminary data with soybean and barley enzymes indicate that this may be a general method for purification of beta-amylases. PMID:11397460
?-Amylase production by Bacillus subtilis with dregs in an external-loop airlift bioreactor
Zheng Yuguo; Wang Zhao; Chen Xiaolong
An external-loop airlift bioreactor, with a low ratio 2.9 of height-to-diameter of the riser and a ratio 6.6 of riser-to-downcomer diameter, was used to produce ?-amylase from fermentation with dregs by Bacillus subtilis. The effects of gas flow rate and liquid volume on ?-amylase production were investigated. After a 36-h fermentation time, an average of 432.3U\\/ml ?-amylase activity was obtained
Ectopic cervical salivary glands
Ph. Greant; M. Pipeleers-Marichal; P. Wylockt
Ectopic salivary glands in the neck are very unusual lesions. Generally they are localized to the oropharyngeal region, near the mandible and para-parotid lymph nodes. They occur occasionally on the anterior border of the sternocleidomastoid muscle in the soft tissues of the neck. We wish to report on a singular association of ectopic salivary glands with partial hearing loss, preauricular
Original article Adaptative significance of amylase
Paris-Sud XI, Université de
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
Amylase: sensitive tumor marker for amylase-producing lung adenocarcinoma.
Zhang, Jie; Zhang, Lixia; Pan, Shiyang; Gu, Bing; Zhen, Yuping; Yan, Jiabin; Zhou, Yiqin
Hyperamylasemia in patients with lung cancer is rarely, comprising 1% to 3% of all lung cancers. This report describes two cases of lung adenocarcinoma coexisting with hyperamylasemia in two women aged 77 and 57, respectively. In these two cases, CT revealed a normal pancreas. We monitored the serum and urine amylase levels during therapy and found it paralleled tumor response to chemotherapy and metastasis. We suggest that the amylase levels are related to the tumor size and might be a valuable factor in predicting chemotherapy and progression of disease for amylase-producing lung cancer. PMID:23991331
Evaluation of alpha- amylase inhibition by Urtica dioica and Juglans regia extracts
Rahimzadeh, Mahsa; Jahanshahi, Samaneh; Moein, Soheila; Moein, Mahmood Reza
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
Optimization of Amylase Production from B. amyloliquefaciens (MTCC 1270) Using Solid State Fermentation
Saha, Koel; Maity, Sujan; Roy, Sudeshna; Pathak, Rishija; Majumdar, Susmita
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
[Amylase inhibitors from Streptomyces lucensis VKPM Ac-1743 and Streptomyces violaceus VKPM Ac-1734].
Sharova, N Iu
Inhibitors synthesized by the Streptomyces lucensis VKPM AS-1743 and Streptomyces violaceus VKPM AS-1734 strains were studied for their influence on amylases of different origin. The effect of the inhibitors was shown to be different on fungal amylase, pancreatic amylase, and amylase from human blood. It has been found that the studied inhibitors are substances of a pseudooligosaccharide nature and exhibit their activity and stability over a wide range of pH and temperature values. The physico-chemical and biochemical properties of isolated inhibitors were compared with those of known microbial inhibitors of ?-glucosidases. PMID:25842903
Cloning and starch degradation profile of maltotriose-producing amylases from Streptomyces species.
Kashiwagi, Norimasa; Miyake, Michiru; Hirose, Shuichi; Sota, Masahiro; Ogino, Chiaki; Kondo, Akihiko
The end products from starch hydrolysis by amylases have important applications in various industries. Here, two amylases derived from two Streptomyces species that hydrolyze soluble starch from potato produced maltotriose as the primary maltooligosaccharide product. The genes, annotated as putative glycoside hydrolases, were cloned and expressed in Streptomyces lividans. These amylases displayed hydrolysis activity from pH 3 to 9.5 and were not affected by Ca(2+.) Optimal production of maltotriose was between 20 and 30 °C at pH 6.5. At the optimal temperature, both amylases produced maltotriose-rich end products rather than either maltose or maltotetraose. PMID:25048235
Affinity column purification of amylases on protein inhibitors from wheat kernel.
Buonocore, V; Poerio, E; Gramenzi, F; Silano, V
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
Purification of alpha-amylases using magnetic alginate beads.
Teotia, S; Gupta, M N
Magnetic alginate beads were used to purify alpha-amylases from porcine pancreas, starchzyme, BAN 240L (a commercial purification from Bacillus subtilis), and wheat germ. The beads bound a significant level of alpha-amylase activity from porcine pancreas, BAN 240L, and wheat germ. In each case, the enzyme activity could be eluted by using 1.0 M maltose, a known competitive inhibitor of alpha-amylase. In the case of BAN 240L, 3.6-fold purification with 72% recovery of activity was observed. In the case of wheat germ enzyme, starting from the crude extract, 48-fold purification with 70% activity recovery was observed. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis also indicated considerable purification in the latter case. PMID:11318034
Children's Context Inappropriate Anger and Salivary Cortisol Robin L. Locke
Wisconsin at Madison, University of
Children's Context Inappropriate Anger and Salivary Cortisol Robin L. Locke University-pituitary-adrenal axis activity, as manifest in salivary cortisol measures. About 23% of the 360 children (ages 6 the hypothesized lower levels of cortisol beyond that attributed to context appropriate anger. Boys' CI anger
Formation of salivary-mucosal pellicle: the role of transglutaminase.
Bradway, S D; Bergey, E J; Scannapieco, F A; Ramasubbu, N; Zawacki, S; Levine, M J
The present investigation was carried out to identify salivary components of mucosal pellicles in vivo and explore further the mechanism of interaction between salivary molecules and buccal epithelial cells. By using specific antisera and immunoprotein blotting, high-(MG1) and low-(MG2) molecular-mass salivary mucins, amylase, salivary cystatins and proline-rich proteins were detected within mucosal pellicle in vivo. In addition, the data indicated that the mucins and proline-rich proteins could be cleaved into lower-molecular-mass products, whereas the proline-rich proteins could also be cross-linked into higher-molecular-mass complexes. The role of buccal epithelial cell transglutaminase in these interactions was further studied by utilizing purified iodinated amylase, neutral cystatin SN and acidic proline-rich proteins 1 and 3 (APRP1 and 3). After incubation with buccal epithelial cells in vitro 125I-labelled APRPs appeared to undergo a greater degree of cross-linking than 125I-labelled cystatin SN, as determined by SDS/PAGE/autoradiography. Amylase did not appear to be cross-linked at all. Recovery of 125I-labelled APRPs and 125I-labelled cystatin SN with epithelial cell envelopes after repeated extraction suggested that both molecules were cross-linked to envelope proteins, but that 125I-labelled APRPs were cross-linked to a greater degree than 125I-labelled cystatin SN. Cross-linking in buccal epithelial cell preparations was inhibited by an excess of methylamine hydrochloride, a transglutaminase substrate. In a further assessment of amylase, cystatin and APRPs as transglutaminase substrates, only APRP3 and a partially purified preparation of APRPs acted as an amine acceptor for the cross-linking of [14C]methylamine by purified transglutaminase, as determined by SDS/PAGE/fluorography. This reaction was completely inhibited by excess EDTA. The combined data from this study suggest that during mucosal pellicle formation multiple components of saliva adsorb to buccal epithelial cell surfaces, and that, within this group, selected components are enzymically cross-linked by an epithelial transglutaminase and/or proteolytically cleaved into smaller fragments. Images Fig 1. Fig 2. Fig 3. Fig 4. Fig 5. Fig 6. Fig 8. PMID:1376115
Formation of salivary-mucosal pellicle: the role of transglutaminase.
Bradway, S D; Bergey, E J; Scannapieco, F A; Ramasubbu, N; Zawacki, S; Levine, M J
The present investigation was carried out to identify salivary components of mucosal pellicles in vivo and explore further the mechanism of interaction between salivary molecules and buccal epithelial cells. By using specific antisera and immunoprotein blotting, high-(MG1) and low-(MG2) molecular-mass salivary mucins, amylase, salivary cystatins and proline-rich proteins were detected within mucosal pellicle in vivo. In addition, the data indicated that the mucins and proline-rich proteins could be cleaved into lower-molecular-mass products, whereas the proline-rich proteins could also be cross-linked into higher-molecular-mass complexes. The role of buccal epithelial cell transglutaminase in these interactions was further studied by utilizing purified iodinated amylase, neutral cystatin SN and acidic proline-rich proteins 1 and 3 (APRP1 and 3). After incubation with buccal epithelial cells in vitro 125I-labelled APRPs appeared to undergo a greater degree of cross-linking than 125I-labelled cystatin SN, as determined by SDS/PAGE/autoradiography. Amylase did not appear to be cross-linked at all. Recovery of 125I-labelled APRPs and 125I-labelled cystatin SN with epithelial cell envelopes after repeated extraction suggested that both molecules were cross-linked to envelope proteins, but that 125I-labelled APRPs were cross-linked to a greater degree than 125I-labelled cystatin SN. Cross-linking in buccal epithelial cell preparations was inhibited by an excess of methylamine hydrochloride, a transglutaminase substrate. In a further assessment of amylase, cystatin and APRPs as transglutaminase substrates, only APRP3 and a partially purified preparation of APRPs acted as an amine acceptor for the cross-linking of [14C]methylamine by purified transglutaminase, as determined by SDS/PAGE/fluorography. This reaction was completely inhibited by excess EDTA. The combined data from this study suggest that during mucosal pellicle formation multiple components of saliva adsorb to buccal epithelial cell surfaces, and that, within this group, selected components are enzymically cross-linked by an epithelial transglutaminase and/or proteolytically cleaved into smaller fragments. PMID:1376115
Further Experiments on Gibberellin-Stimulated Amylase Production in Cereal Grains
ERIC Educational Resources Information Center
Coppage, Jo; Hill, T. A.
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)
An efficient purification process for sweet potato beta-amylase by affinity precipitation with alginate
Sunita Teotia; S. K Khare; M. N Gupta
?-amylases are used in production of maltose syrup. It is shown that sweet potato ?-amylase can be purified by affinity precipitation with alginate with 80% activity yield and 44 fold purification. SDS-PAGE of the purified protein showed a single band and a subunit weight of 50 kDa. Preliminary data with soybean and barley enzymes indicate that this may be a
Salivary mucoepidermoid carcinoma revisited.
Coca-Pelaz, Andrés; Rodrigo, Juan P; Triantafyllou, Asterios; Hunt, Jennifer L; Rinaldo, Alessandra; Strojan, Primož; Haigentz, Missak; Mendenhall, William M; Takes, Robert P; Vander Poorten, Vincent; Ferlito, Alfio
Clinicopathological features, prognosis and therapeutic strategies for mucoepidermoid carcinoma originating in salivary and salivary-type glands of the head and neck are reviewed. We emphasise histopathological aspects, appraise the value of histochemistry, electron microscopy, immunohistochemistry and cytophotometry, and discuss histogenesis and characteristic gene translocations. We additionally consider possible diagnostic difficulties, problems related to histological grading and accuracy of existing literature, and areas of controversy or uncertainty which may benefit from further investigations. PMID:24771140
Analysis of the Salivary Gland Transcriptome of Frankliniella occidentalis
Stafford-Banks, Candice A.; Rotenberg, Dorith; Johnson, Brian R.; Whitfield, Anna E.; Ullman, Diane E.
Saliva is known to play a crucial role in insect feeding behavior and virus transmission. Currently, little is known about the salivary glands and saliva of thrips, despite the fact that Frankliniella occidentalis (Pergande) (the western flower thrips) is a serious pest due to its destructive feeding, wide host range, and transmission of tospoviruses. As a first step towards characterizing thrips salivary gland functions, we sequenced the transcriptome of the primary salivary glands of F. occidentalis using short read sequencing (Illumina) technology. A de novo-assembled transcriptome revealed 31,392 high quality contigs with an average size of 605 bp. A total of 12,166 contigs had significant BLASTx or tBLASTx hits (E?1.0E?6) to known proteins, whereas a high percentage (61.24%) of contigs had no apparent protein or nucleotide hits. Comparison of the F. occidentalis salivary gland transcriptome (sialotranscriptome) against a published F. occidentalis full body transcriptome assembled from Roche-454 reads revealed several contigs with putative annotations associated with salivary gland functions. KEGG pathway analysis of the sialotranscriptome revealed that the majority (18 out of the top 20 predicted KEGG pathways) of the salivary gland contig sequences match proteins involved in metabolism. We identified several genes likely to be involved in detoxification and inhibition of plant defense responses including aldehyde dehydrogenase, metalloprotease, glucose oxidase, glucose dehydrogenase, and regucalcin. We also identified several genes that may play a role in the extra-oral digestion of plant structural tissues including ?-glucosidase and pectin lyase; and the extra-oral digestion of sugars, including ?-amylase, maltase, sucrase, and ?-glucosidase. This is the first analysis of a sialotranscriptome for any Thysanopteran species and it provides a foundational tool to further our understanding of how thrips interact with their plant hosts and the viruses they transmit. PMID:24736614
Technology Transfer Automated Retrieval System (TEKTRAN)
Using specific proteinase inhibitors, we demonstrated that serine proteinases in the tarnished plant bug, Lygus lineolaris, are major proteinases in both salivary glands and gut tissues. Gut proteinases were less sensitive to inhibition than proteinases from the salivary glands. Up to 80% azocaseina...
Zinc oxide nanoparticles as novel alpha-amylase inhibitors
NASA Astrophysics Data System (ADS)
Dhobale, Sandip; Thite, Trupti; Laware, S. L.; Rode, C. V.; Koppikar, Soumya J.; Ghanekar, Ruchika-Kaul; Kale, S. N.
Amylase inhibitors, also known as starch blockers, contain substances that prevent dietary starches from being absorbed by the body via inhibiting breakdown of complex sugars to simpler ones. In this sense, these materials are projected as having potential applications in diabetes control. In this context, we report on zinc oxide nanoparticles as possible alpha-amylase inhibitors. Zinc oxide nanoparticles have been synthesized using soft-chemistry approach and 1-thioglycerol was used as a surfactant to yield polycrystalline nanoparticles of size ˜18 nm, stabilized in wurtzite structure. Conjugation study and structural characterization have been done using x-ray diffraction technique, Fourier transform infrared spectroscopy, UV-visible spectroscopy, and transmission electron microscopy. Cytotoxicity studies on human fibrosarcoma (HT-1080) and skin carcinoma (A-431) cell lines as well as mouse primary fibroblast cells demonstrate that up to a dose of 20 ?g/ml, ZnO nanoparticles are nontoxic to the cells. We report for the first time the alpha-amylase inhibitory activity of ZnO nanoparticles wherein an optimum dose of 20 ?g/ml was sufficient to exhibit 49% glucose inhibition at neutral pH and 35 °C temperature. This inhibitory activity was similar to that obtained with acarbose (a standard alpha-amylase inhibitor), thereby projecting ZnO nanoparticles as novel alpha-amylase inhibitors.
Site-directed mutagenesis of histidine 93, aspartic acid 180, glutamic acid 205, histidine 290, and aspartic acid 291 at the active site and tryptophan 279 at the raw starch binding site in barley alpha-amylase 1.
Søgaard, M; Kadziola, A; Haser, R; Svensson, B
The pseudotetrasaccharide acarbose has high affinity for the active site (Ki,app = 1 microM) and low affinity for a secondary site (Kd = 2.3 mM) in barley alpha-amylase 1, distinguished by inhibition kinetics and spectral perturbation. Mutants of putative catalytic residues, D180N, E205Q, and D291N, are inactive and display low affinity for acarbose-Sepharose. H93N and H290N mutants, at invariant residues, have kcat/Km for p-nitrophenylmaltoheptaoside of 0.3 and 1.2% of wild-type. A corresponding 370- and 85-fold increased Ki,app for acarbose and a lack of shifts in pH activity profiles indicate that these histidines participate in transition state stabilization but not directly in catalysis. This finding agrees with H bonding to OH groups of the valienamine ring of acarbose in the three-dimensional structure. Loss of inhibition above pH 6 supports that acarbose is most potent in protonated form. The low affinity site contains Trp278 and Trp279, known to bind cyclomaltoheptaose. While the W279A mutant has 10-fold decreased affinity for starch granules, production of Trp278 mutants failed. The invariant Trp278 is perhaps critical for stability or folding in cereal alpha-amylases. PMID:7901200
Amylase production by endophytic fungi Cylindrocephalumsp. isolated from medicinal plant Alpinia calcarata (Haw.) Roscoe
Sunitha, V. H.; Ramesha, A.; Savitha, J.; Srinivas, C
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
What Is Salivary Gland Cancer?
... a slightly better outcome. Other cancers that can affect the salivary glands These types of cancer are ... Most lymphomas that start in the salivary glands affect people with Sjogren (Sjögren) syndrome (a disorder that ...
Expression and Characterization of Geobacillus stearothermophilus SR74 Recombinant ?-Amylase in Pichia pastoris
Gandhi, Sivasangkary; Salleh, Abu Bakar; Rahman, Raja Noor Zaliha Raja Abd; Chor Leow, Thean; Oslan, Siti Nurbaya
Geobacillus stearothermophilus SR74 is a locally isolated thermophilic bacteria producing thermostable and thermoactive ?-amylase. Increased production and commercialization of thermostable ?-amylase strongly warrant the need of a suitable expression system. In this study, the gene encoding the thermostable ?-amylase in G. stearothermophilus SR74 was amplified, sequenced, and subcloned into P. pastoris GS115 strain under the control of a methanol inducible promoter, alcohol oxidase (AOX). Methanol induced recombinant expression and secretion of the protein resulted in high levels of extracellular amylase production. YPTM medium supplemented with methanol (1% v/v) was the best medium and once optimized, the maximum recombinant ?-amylase SR74 achieved in shake flask was 28.6?U?mL?1 at 120?h after induction. The recombinant 59?kDa ?-amylase SR74 was purified 1.9-fold using affinity chromatography with a product yield of 52.6% and a specific activity of 151.8?U?mg?1. The optimum pH of ?-amylase SR74 was 7.0 and the enzyme was stable between pH 6.0–8.0. The purified enzyme was thermostable and thermoactive, exhibiting maximum activity at 65°C with a half-life (t1/2) of 88?min at 60°C. In conclusion, thermostable ?-amylase SR74 from G. stearothermophilus SR74 would be beneficial for industrial applications, especially in liquefying saccrification.
Alpha-amylase from the Hyperthermophilic Archaeon Thermococcus thioreducens
NASA Technical Reports Server (NTRS)
Bernhardsdotter, E. C. M. J.; Pusey, M. L.; Ng, M. L.; Garriott, O. K.
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.
Properties of an amylase from thermophilic Bacillus SP
de Carvalho, Raquel Vieira; Côrrea, Thamy Lívia Ribeiro; da Silva, Júlia Caroline Matos; de Oliveira Mansur, Luciana Ribeiro Coutinho; Martins, Meire Lelis Leal
?-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
Production and Partial Purification of Alpha Amylase from Bacillus subtilis (MTCC 121) Using Solid State Fermentation
Raul, Dibyangana; Mukhopadhyay, Suchita; Kumar Das, Shrayan; Gupta, Suvroma
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
Salivary glands and human selection: a hypothesis.
Shields, E D; Mann, R W
Stafne static bone defect (SSBD) of the mandible is the only described destructive bone lesion that is highly localized, nonprogressive, but nonhealing. This common defect in male is found in the region of the major salivary glands that produce a cornucopia of biologically active factors. We describe rare phenocopies caused by mandibular immobility that hold the gland in a constant position thus implicating a localized chronic "leak" of an osteoclast induction factor from the major salivary glands as the pathologic agent. This finding suggests that increased salivary gland size could simulate immobility by apposing the gland to bone, thus allowing the "leaked" factor's gradient to have an effect. In one step, the putative genetic enlargement of a critical gland that produces many factors important for survival, a broad biological vista would be available to the massive potential for both positive and negative selection. Positive selection was identified by observing a correlation between the prevalence of enhanced androgen-induced enlarge salivary glands (SSBD) as a marker, with a great preponderance of males) and the conjectured resulting increased production of immunoreactive factors, with pole-to-equator isotherm and broad ranged infection clines. Negative selection was observed among the slave ancestors of African Americans for a potential embryonic homeotic mutation causing larger salivary glands in both sexes (decreased prevalence of SSBD, with an equal sex ratio). The decreased production of saliva and electrolytes diminished the salt and water depletive effects of severe diarrhea and vomiting induced by enteric diseases, which killed many slaves. Data presented suggests that SSBD is a polymorphism and a marker of selection processes that cause variation in size, or structure, of the major salivary glands. PMID:8773904
Prevalence of the Amylase-Binding Protein A Gene (abpA) in Oral Streptococci
Brown, Alan E.; Rogers, Jeffrey D.; Haase, Elaine M.; Zelasko, Peter M.; Scannapieco, Frank A.
Salivary amylase binds specifically to a number of oral streptococcal species. This interaction may play an important role in dental plaque formation. Recently, a 585-bp gene was cloned and sequenced from Streptococcus gordonii Challis encoding a 20.5-kDa amylase-binding protein (AbpA). The goal of this study was to determine if related genes are present in other species of oral streptococci. Biotinylated abpA was used in Southern blot analysis to screen genomic DNA from several strains representing eight species of oral streptococci. This probe hybridized with a 4.0-kb HindIII restriction fragment from all 13 strains of S. gordonii tested. The probe did not appear to bind to any restriction fragments from other species of amylase-binding oral streptococci including Streptococcus mitis (with the exception of 1 of 14 strains), Streptococcus crista (3 strains), Streptococcus anginosus (1 strain), and Streptococcus parasanguinis (1 strain), or to non-amylase-binding oral streptococci including Streptococcus sanguinis (3 strains), Streptococcus oralis (4 strains), and Streptococcus mutans (1 strain). Primers homologous to sequences within the 3? and 5? ends of abpA yielded products of 400 bp following PCR of genomic DNA from the Southern blot-positive strains. Several of these PCR products were cloned and sequenced. The levels of similarity of these cloned products to the abpA of S. gordonii Challis ranged from 91 to 96%. These studies reveal that the abpA gene appears to be specific to S. gordonii and differs from genes encoding amylase-binding proteins from other species of amylase-binding streptococci. PMID:10565935
Prevalence of the amylase-binding protein A gene (abpA) in oral streptococci.
Brown, A E; Rogers, J D; Haase, E M; Zelasko, P M; Scannapieco, F A
Salivary amylase binds specifically to a number of oral streptococcal species. This interaction may play an important role in dental plaque formation. Recently, a 585-bp gene was cloned and sequenced from Streptococcus gordonii Challis encoding a 20.5-kDa amylase-binding protein (AbpA). The goal of this study was to determine if related genes are present in other species of oral streptococci. Biotinylated abpA was used in Southern blot analysis to screen genomic DNA from several strains representing eight species of oral streptococci. This probe hybridized with a 4.0-kb HindIII restriction fragment from all 13 strains of S. gordonii tested. The probe did not appear to bind to any restriction fragments from other species of amylase-binding oral streptococci including Streptococcus mitis (with the exception of 1 of 14 strains), Streptococcus crista (3 strains), Streptococcus anginosus (1 strain), and Streptococcus parasanguinis (1 strain), or to non-amylase-binding oral streptococci including Streptococcus sanguinis (3 strains), Streptococcus oralis (4 strains), and Streptococcus mutans (1 strain). Primers homologous to sequences within the 3' and 5' ends of abpA yielded products of 400 bp following PCR of genomic DNA from the Southern blot-positive strains. Several of these PCR products were cloned and sequenced. The levels of similarity of these cloned products to the abpA of S. gordonii Challis ranged from 91 to 96%. These studies reveal that the abpA gene appears to be specific to S. gordonii and differs from genes encoding amylase-binding proteins from other species of amylase-binding streptococci. PMID:10565935
Establishment of functional acinar-like cultures from human salivary glands.
Jang, S I; Ong, H L; Gallo, A; Liu, X; Illei, G; Alevizos, I
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
Assessment of Salivary Hormones
Schultheiss, Oliver C.
on an individual's sex hormones (e.g., Graham & Desjardins, 1980; Roney, Lukaszewski, & Simmons, 2007); or when17 3 Assessment of Salivary Hormones Oliver C. Schultheiss Steven J. Stanton A Primer on Concepts episodic memory when it binds to receptors in the brain (e.g., Beckwith, Petros, Bergloff, & Staebler, 1987
alpha-Amylase immobilized on bulk acoustic-wave sensor by UV-curing coating.
He; Cai; Wei; Nie; Yao
A new method for immobilization of alpha-amylase by UV-curing coating is proposed in this paper. The immobilization procedure of UV-curing coating on piezoelectric quartz crystal is simple and convenient, and causes less loss of enzymatic activity. The activity of the immobilized alpha-amylase is monitored by a technique based on bulk acoustic-wave (BAW) sensor. The frequency shift of BAW sensor can reflect the degree of hydrolysis of starch by the immobilized alpha-amylase. It is appropriate for the immobilized alpha-amylase to hydrolyze the soluble starch under pH 7.0 condition, which is similar to that of the free alpha-amylase. Kinetic parameters (the Michaelis constant, K(m), and the maximum initial rate V(max)) of the enzymatic hydrolysis of starch by the immobilized alpha-amylase are estimated by using a linear method of Lineweaver-Burk plot. K(m)=12.7mgml(-1) and V(max)=15.9Hzmin(-1). And the experimental results show that the immobilized alpha-amylase entrapped by the UV-curing coating retains adequate enzymatic activity and can be reused more than 50 times under certain experimental conditions. PMID:10908862
Determination of the in vitro and in vivo antimicrobial activity on salivary Streptococci and Lactobacilli and chemical characterisation of the phenolic content of a Plantago lanceolata infusion.
Ferrazzano, Gianmaria Fabrizio; Cantile, Tiziana; Roberto, Lia; Ingenito, Aniello; Catania, Maria Rosaria; Roscetto, Emanuela; Palumbo, Giuseppe; Zarrelli, Armando; Pollio, Antonino
Introduction. Plant extracts may be suitable alternative treatments for caries. Aims. To investigate the in vitro and in vivo antimicrobial effects of Plantago lanceolata herbal tea (from flowers and leaves) on cariogenic bacteria and to identify the major constituents of P. lanceolata plant. Materials and Methods. The MIC and MBC against cariogenic bacteria were determined for P. lanceolata tea. Subsequently, a controlled random clinical study was conducted. Group A was instructed to rinse with a P. lanceolata mouth rinse, and Group B received a placebo mouth rinse for seven days. The salivary colonisation by streptococci and lactobacilli was investigated prior to treatment and on the fourth and seventh days. Finally, the P. lanceolata tea was analysed for its polyphenolic content, and major phenolics were identified. Results and Discussion. P. lanceolata teas demonstrate good in vitro antimicrobial activity. The in vivo test showed that Group A subjects presented a significant decrease in streptococci compared to Group B. The phytochemical analysis revealed that flavonoids, coumarins, lipids, cinnamic acids, lignans, and phenolic compounds are present in P. lanceolata infusions. Conclusions. P. lanceolata extract could represent a natural anticariogenic agent via an antimicrobial effect and might be useful as an ancillary measure to control the proliferation of cariogenic flora. PMID:25767805
Determination of the In Vitro and In Vivo Antimicrobial Activity on Salivary Streptococci and Lactobacilli and Chemical Characterisation of the Phenolic Content of a Plantago lanceolata Infusion
Roberto, Lia; Ingenito, Aniello; Roscetto, Emanuela
Introduction. Plant extracts may be suitable alternative treatments for caries. Aims. To investigate the in vitro and in vivo antimicrobial effects of Plantago lanceolata herbal tea (from flowers and leaves) on cariogenic bacteria and to identify the major constituents of P. lanceolata plant. Materials and Methods. The MIC and MBC against cariogenic bacteria were determined for P. lanceolata tea. Subsequently, a controlled random clinical study was conducted. Group A was instructed to rinse with a P. lanceolata mouth rinse, and Group B received a placebo mouth rinse for seven days. The salivary colonisation by streptococci and lactobacilli was investigated prior to treatment and on the fourth and seventh days. Finally, the P. lanceolata tea was analysed for its polyphenolic content, and major phenolics were identified. Results and Discussion. P. lanceolata teas demonstrate good in vitro antimicrobial activity. The in vivo test showed that Group A subjects presented a significant decrease in streptococci compared to Group B. The phytochemical analysis revealed that flavonoids, coumarins, lipids, cinnamic acids, lignans, and phenolic compounds are present in P. lanceolata infusions. Conclusions. P. lanceolata extract could represent a natural anticariogenic agent via an antimicrobial effect and might be useful as an ancillary measure to control the proliferation of cariogenic flora. PMID:25767805
Purification of extrachloroplastic. beta. -amylase from leaves of starchless and wild type Arabidopsis
Somerville, C.; Monroe, J.; Preiss, J. (Michigan Sate Univ., East Lansing (USA))
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.
21 CFR 184.1012 - ?-Amylase enzyme preparation from Bacillus stearothermophilus.
Code of Federal Regulations, 2012 CFR
...stearothermophilus. (a) ?-Amylase enzyme preparation is obtained from the culture filtrate that results from a pure culture fermentation of a nonpathogenic and nontoxicogenic strain of Bacillus stearothermophilus. Its characterizing enzyme activity is...
21 CFR 184.1012 - ?-Amylase enzyme preparation from Bacillus stearothermophilus.
Code of Federal Regulations, 2013 CFR
...stearothermophilus. (a) ?-Amylase enzyme preparation is obtained from the culture filtrate that results from a pure culture fermentation of a nonpathogenic and nontoxicogenic strain of Bacillus stearothermophilus. Its characterizing enzyme activity is...
Salivary defense system alters in vegetarian
Amirmozafari, Nour; Pourghafar, Houra; Sariri, Reyhaneh
Purpose The aim of this research was investigating antimicrobial and enzymatic antioxidant activities in salivary fluids of vegetarians as compared to normal subjects. Material & Methods Antimicrobial activity of the saliva samples was evaluated against four clinically important bacteria. The biological activities of three of the main antioxidant enzymes of saliva were measured using appropriate methods of enzyme assay in both groups. Results According to the results, saliva obtained from vegetarians showed a reduced inhibitory effect on growth of Staphylococcus aureus, Klebsiella oxytoca, Pseudomonas aeruginosa and Escherichia coli as compared to those obtained from the non-vegetarian subjects. The activity of salivary peroxidase, catalase and superoxide dismutase showed a statistically marked decrease in vegetarian group. Conclusions According to our literature survey, this is the first report on the antibacterial and antioxidant capacity in saliva of vegetarians. Results obtained from the present study have opened a new line of research with the basis of saliva as a research tool. PMID:25737889
Characteristics of Two Forms of ?-Amylases and Structural Implication
Ohdan, Kohji; Kuriki, Takashi; Kaneko, Hiroki; Shimada, Jiro; Takada, Toshikazu; Fujimoto, Zui; Mizuno, Hiroshi; Okada, Shigetaka
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
Purification and Partial Characterization of Proteinase and ?-Amylase Inhibitors from Lesser Yam (Dioscorea esculenta)
K. Sasikiran; M. R. Rekha; G. Padmaja
The proteinase and ?-amylase isoinhibitors of lesser yam were isolated through TCA precipitation and DEAE cellulose chromatography. The major peak with proteinase inhibitor activity obtained by DEAE-cellulose chromatography was further fractionated to five isoinhibitor fractions with molecular weights 67, 50, 27, 15, and 12 kDa respectively in GPC and PAGE. On the contrary, the major peak with ?-amylase inhibitor activity segregated
Beta-amylase in germinating millet seeds.
Yamasaki, Yoshiki
Beta-amylase (EC 126.96.36.199) 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
A simple one pot purification of bacterial amylase from fermented broth based on affinity toward starch-functionalized magnetic nanoparticle.
Paul, Tanima; Chatterjee, Saptarshi; Bandyopadhyay, Arghya; Chattopadhyay, Dwiptirtha; Basu, Semanti; Sarkar, Keka
Surface-functionalized adsorbant particles in combination with magnetic separation techniques have received considerable attention in recent years. Selective manipulation on such magnetic nanoparticles permits separation with high affinity in the presence of other suspended solids. Amylase is used extensively in food and allied industries. Purification of amylase from bacterial sources is a matter of concern because most of the industrial need for amylase is met by microbial sources. Here we report a simple, cost-effective, one-pot purification technique for bacterial amylase directly from fermented broth of Bacillus megaterium utilizing starch-coated superparamagnetic iron oxide nanoparticles (SPION). SPION was prepared by co-precipitation method and then functionalized by starch coating. The synthesized nanoparticles were characterized by transmission electron microscopy (TEM), a superconducting quantum interference device (SQUID, zeta potential, and ultraviolet-visible (UV-vis) and Fourier-transform infrared (FTIR) spectroscopy. The starch-coated nanoparticles efficiently purified amylase from bacterial fermented broth with 93.22% recovery and 12.57-fold purification. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the molecular mass of the purified amylase was 67 kD, and native gel showed the retention of amylase activity even after purification. Optimum pH and temperature of the purified amylase were 7 and 50°C, respectively, and it was stable over a range of 20°C to 50°C. Hence, an improved one-pot bacterial amylase purification method was developed using starch-coated SPION. PMID:24840788
Impact of ?-Amylases on Quality Characteristics of Asian White Salted Noodles Made From Australian White Wheat Flour
Larisa Cato; Andrew L. Halmos; Darryl M. Small
Cereal Chem. 83(5):491-497 The consumer acceptance of white salted Asian noodles depends on starch characteristics, and the purpose of this study has been to investi- gate the potential of exogenous ?-amylases to enhance textural charac- teristics of this product. Noodles were prepared from commercial flours with low ?-amylase activity, and the endogenous enzyme remained rela- tively stable during various processing
endoProteoFASP: a novel FASP approach to profile salivary peptidome and disclose salivary proteases.
Trindade, Fábio; Amado, Francisco; Gomes, Pedro S; Vitorino, Rui
The salivary peptidome, which can represent up to 20% of total secreted proteins in human saliva, is highly influenced by proteolytic events. However, the development of strategies to understand the dynamics underlying the generation of salivary peptides has been a challenging task. In order to disclose in more detail the proteolytic events taking place in saliva, we aimed to characterize salivary peptidome and predict salivary proteases by applying, for the first time, a filter-aided sample preparation (FASP) approach to saliva. Thus, as a proof-of-concept of this application, harvested saliva samples from healthy individuals were incubated in 30 kDa cut-off spin filters for 18 or 115 h, at 37 °C, to promote saliva autolysis and the attained peptidome was characterized and compared with the naturally occurring one. In ex vivo conditions, proline-rich proteins, P-B peptide, histatin 1 and statherin were found to be the most susceptible salivary proteins to proteolysis. Peptide fragments were mainly attributed to the activity of cathepsin L1 and K at 18 h, whereas at 115 h, the attained peptide fragments were attributed to the activity of cathepsins K and L1, and MEP1A. Overall, the described endoProteoFASP approach makes the most of saliva?s own protease pool and avoids the use of synthetic peptides and exogenous proteases to understand the proteolytic events occurring in the oral fluid. Hence, it could be very helpful in future studies targeting the characterization of salivary proteases and peptidome from different pathophysiological conditions. PMID:25476335
An exceptionally cold-adapted alpha-amylase from a metagenomic library of a cold and alkaline environment.
Vester, Jan Kjølhede; Glaring, Mikkel Andreas; Stougaard, Peter
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
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...
21 CFR 862.1070 - Amylase test system.
Code of Federal Regulations, 2014 CFR
...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...
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...
Effect of modifying histidine residues on the action of Bacillus amyloliquefaciens and barley-malt alpha-amylases.
Nakatani, H; Hamaguchi, K; Ishikawa, K
Modification of porcine pancreatic alpha-amylase (PPA) and Taka-amylase A(TAA) with diethyl pyrocarbonate (DEP) causes activation of the release of p-nitrophenol from p-nitrophenol alpha-maltoside (G2PNP), and a decrease in amylase activity (hydrolysis of alpha-1,4 glucosidic bonds in starch). Among the possible sites of modification, attention focuses on three histidine residues present around the active site of alpha-amylases of many different origins. In PPA these are His 101, His 201, and His 299, with His 101 and His 299 being very close to the site of catalysis and thus perhaps directly or indirectly involved in the catalysis. On the other hand, His 201 is located on the aglycon side of the catalytic site, and we have suggested that it is involved in the increase of PNP release after chemical modification. Investigations of site-directed mutagenesis of the histidine residues of human pancreatic alpha-amylase support this identification. Although the degree of sequence similarity among alpha-amylases of different origins is low, there are several conserved short regions. Most belong to the structural components of the active site in PPA and TAA. Furthermore, there is a close similarity in the three-dimensional structures of PPA and TAA. The conserved residues around the active site in all alpha-amylases suggest some universal structural similarities in these active sites. Therefore, we examined the effects of the chemical modification of histidine residues in Bacillus amyloliquefaciens alpha-amylase(BLA) with DEP and made a comparison with modification of barley alpha-amylase isoenzyme II(BAII), using identical substrate systems. These two alpha-amylases have more substrate binding subsites than PPA and TAA, and have similar action patterns with malto-oligosaccharides. PMID:8004636
Acute salivary gland hypofunction in the duct ligation model in the absence of inflammation
Correia, PN; Carpenter, GH; Osailan, SM; Paterson, KL; Proctor, GB
Objective The commonly associated aetiology of salivary gland inflammation and salivary hypofunction has led to the widely held belief that inflammation causes salivary gland hypofunction. Indeed, our own recent study seemed to support this contention. Here, we tested the hypothesis that, in an acute duct ligation model, eliminating inflammation the submandibular gland would recover normal function. Materials and methods Ligation of the rat submandibular gland excretory duct for 24 h was used to induce inflammation and salivary gland hypofunction. A group of duct ligated rats was compared with a second group given dexamethasone, on the day of duct ligation. Twenty-four hours later salivary gland function was assessed and salivary glands were collected. Results Histology and myeloperoxidase activity assay revealed a profound decrease in inflammatory cell infiltration of ligated glands from rats given dexamethasone, compared with ligated glands in the absence of dexamethasone. Salivary flow rate evoked by methacholine was decreased (P < 0.01) by approximately 56% (ligated vs control, 79 ± 9 ?l min?1 g?1vs 177 ± 11 ?l min?1 g?1) and salivary flow from ligated dexamethasone-treated and ligated glands was similar. Conclusion Despite eliminating the inflammatory reaction in the ligated gland, salivary hypofunction was not reversed, suggesting that other mechanisms must be at work in the ligation-induced salivary hypofunction. PMID:18221457
Acid diffusion into rice boluses is influenced by rice type, variety, and presence of ?-amylase.
Mennah-Govela, Yamile A; Bornhorst, Gail M; Singh, R Paul
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
Dasatinib in Treating Patients With Recurrent or Metastatic Malignant Salivary Gland Tumors
High-grade Salivary Gland Mucoepidermoid Carcinoma; Low-grade Salivary Gland Mucoepidermoid Carcinoma; Recurrent Salivary Gland Cancer; Salivary Gland Acinic Cell Tumor; Salivary Gland Adenocarcinoma; Salivary Gland Adenoid Cystic Carcinoma; Salivary Gland Anaplastic Carcinoma; Salivary Gland Malignant Mixed Cell Type Tumor; Salivary Gland Poorly Differentiated Carcinoma; Salivary Gland Squamous Cell Carcinoma; Stage IV Salivary Gland Cancer
Engineering ?-amylase levels in wheat grain suggests a highly sophisticated level of carbohydrate regulation during development.
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
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
Engineering ?-amylase levels in wheat grain suggests a highly sophisticated level of carbohydrate regulation during development
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
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
Immunohistochemical analysis of CD34 expression in salivary gland tumors
Moghadam, Saede Atarbashi; Abadi, Ayda Mohammad; Mokhtari, Sepideh
Background: Tumor growth depends on angiogenesis which is assessed by measuring the tumor microvessel density (MVD) through CD34 immunostaining. The present study was performed to evaluate the situation of angiogenic activity in salivary gland neoplasms. The possible role of CD34 in progression and invasion of salivary gland tumors is also investigated. Materials and Methods: Tissue specimens of 15 pleomorphic adenoma (PA) and 15 malignant salivary gland tumors including mucoepidermoid carcinoma (MEC), adenoid cystic carcinoma (AdCC) and salivary duct carcinoma (SDC) were immunostained for CD34 protein. The most vascularized areas at low power magnification (hotspots) were selected for vessel counting at ×400 magnification. Then, the mean number of microvessels in three fields within the tumor mass was calculated. Results: MVD in PA and malignant salivary gland tumors were 10.93 ± 5.95 and 26.46 ± 7.32, respectively. Tumor angiogenesis in PA was much lower than other lesions (P < 0.05). No significant difference was observed between malignant tumors (P > 0.05). Conclusion: Salivary gland carcinomas demonstrated higher vascular density than benign PA despite of cell types and architecture. The reason for this higher angiogenic activity could be related to metabolic characteristics of malignant cells.
Production and partial purification of ?-amylase from a novel isolate Streptomyces gulbargensis
Dastager G. Syed; Dayanand Agasar; Ashok Pandey
Extracellular amylase production by a newly isolated alkali-thermotolerant strain Streptomyces gulbargensis DAS 131 was optimized and characterized. The highest amylase production was achieved by growing S. gulbargensis DAS 131 in media with 1% starch. Strain exhibited maximal activity at pH 9.0 and 45°C and relatively stable in alkaline conditions\\u000a (pH 11). Starch and peptone were found to be the good
Chronic activation of the epithelial immune system of the fruit fly's salivary glands has a negative effect on organismal growth and induces a peculiar set of target genes
Ahmed Abdelsadik; Thomas Roeder
BACKGROUND: Epithelial and especially mucosal immunity represents the first line of defence against the plethora of potential pathogens trying to invade via the gastrointestinal tract. The salivary glands of the fruit fly are an indispensable part of the gastrointestinal tract, but their contribution to the mucosal immunity has almost completely been neglected. Our major goal was to elucidate if the
Salivary gland diseases in children
Iro, Heinrich; Zenk, Johannes
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
Physical activity, job demand–control, perceived stress–energy, and salivary cortisol in white-collar workers
Åse Marie Hansen; Anne Katrine Blangsted; Ernst Albin Hansen; Karen Søgaard; Gisela Sjøgaard
Purpose The aim of the present study is to examine the association between physical activity and perceived job demand, job control,\\u000a perceived stress and energy, and physiological arousal reflected by morning and evening concentrations of cortisol in saliva\\u000a among white-collar workers.\\u000a \\u000a \\u000a \\u000a Methods Physical activity during the last week was assessed during work and leisure time by a Danish version of the International
Production of itaconic acid in Escherichia coli expressing recombinant ?-amylase using starch as substrate.
Okamoto, Shusuke; Chin, Taejun; Nagata, Keisuke; Takahashi, Tetsuya; Ohara, Hitomi; Aso, Yuji
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
Control of alpha-Amylase Development in Cotyledons during and following Germination of Mung Bean Seeds.
Morohashi, Y; Katoh, H; Kaneko, Y; Matsushima, H
Developmental patterns of alpha-amylase in Vigna radiata cotyledons during and following germination were quite different depending on the differences in the treatments of cotyledons during the imbibitional stage. When axis-detached cotyledons were imbibed in water with seed-coats attached, alpha-amylase activity did not increase and remained low. On the other hand, when the cotyledons were imbibed in water after seed-coat removal, the enzyme activity increased markedly. If the axis was attached to the cotyledons, alpha-amylase showed a marked development even under the former imbibition conditions. These changes in the enzyme activity were in parallel with those in the enzyme content, and the content, in turn, was dependent upon the availability of mRNA for alpha-amylase. We propose that the regulation of the development of alpha-amylase in cotyledons may involve some factor(s) inhibitory to accumulation of alpha-amylase mRNA, which is present in dry cotyledons and can be removed from cotyledons by leakage or by the presence of the axis. PMID:16667006
Environmental and Genetic Contributors to Salivary Testosterone Levels in Infants
Xia, Kai; Yu, Yang; Ahn, Mihye; Zhu, Hongtu; Zou, Fei; Gilmore, John H.; Knickmeyer, Rebecca C.
Transient activation of the hypothalamic–pituitary–gonadal axis in early infancy plays an important role in male genital development and sexual differentiation of the brain, but factors contributing to individual variation in testosterone levels during this period are poorly understood. We measured salivary testosterone levels in 222 infants (119 males, 103 females, 108 singletons, 114 twins) between 2.70 and 4.80?months of age. We tested 16 major demographic and medical history variables for effects on inter-individual variation in salivary testosterone. Using the subset of twins, we estimated genetic and environmental contributions to salivary testosterone levels. Finally, we tested single nucleotide polymorphisms (SNPs) within ±5?kb of genes involved in testosterone synthesis, transport, signaling, and metabolism for associations with salivary testosterone using univariate tests and random forest (RF) analysis. We report an association between 5?min APGAR scores and salivary testosterone levels in males. Twin modeling indicated that individual variability in testosterone levels was primarily explained by environmental factors. Regarding genetic variation, univariate tests did not reveal any variants significantly associated with salivary testosterone after adjusting for false discovery rate. The top hit in males was rs10923844, an SNP of unknown function located downstream of HSD3B1 and HSD3B2. The top hits in females were two SNPs located upstream of ESR1 (rs3407085 and rs2295190). RF analysis, which reflects joint and conditional effects of multiple variants, indicated that genes involved in regulation of reproductive function, particularly LHCGR, are related to salivary testosterone levels in male infants, as are genes involved in cholesterol production, transport, and removal, while genes involved in estrogen signaling are related to salivary testosterone levels in female infants. PMID:25400620
On the origin and diagnostic use of salivary RNA.
Fábryová, H; Celec, P
Saliva as a diagnostic fluid enables non-invasive sampling, which can be performed even by an untrained person. Saliva is, thus, particularly useful for large population screenings, for children, elderly and whenever repeated samplings are needed. Saliva is a plasma filtrate actively modified by the salivary glands. Saliva could replace some routine blood tests in the future. The sources of salivary RNA include oral epithelial cells and oral micro-organisms. Recent developments suggest that using known salivary RNA markers, it is possible to diagnose diseases such as oral carcinoma and other diseases will be added soon. Salivary RNA can be used to identify oral bacteria and to determine the expression of specific genes. On a systemic level, it provides information about the whole oral transcriptome and microbiome. Despite the small amount of salivary RNA, the issues with its isolation have been overcome. Saliva, thus, contains RNA of sufficient quality and quantity for sensitive and specific analyses. Salivary RNA can provide medically relevant information about oral microbiome, oral carcinoma, but also breast and pancreatic cancer and is, thus, a promising tool for future research and clinical diagnostics. PMID:23517132
Silk fibroin scaffolds promote formation of the ex vivo niche for salivary gland epithelial cell growth, matrix formation, and retention of differentiated function.
Zhang, Bin-Xian; Zhang, Zhi-Liang; Lin, Alan L; Wang, Hanzhou; Pilia, Marcello; Ong, Joo L; Dean, David D; Chen, Xiao-Dong; Yeh, Chih-Ko
Salivary gland hypofunction often results from a number of causes, including the use of various medications, radiation for head and neck tumors, autoimmune diseases, diabetes, and aging. Since treatments for this condition are lacking and adult salivary glands have little regenerative capacity, there is a need for cell-based therapies to restore salivary gland function. Development of these treatment strategies requires the establishment of a system that is capable of replicating the salivary gland cell "niche" to support the proliferation and differentiation of salivary gland progenitor cells. In this study, a culture system using three-dimensional silk fibroin scaffolds (SFS) and primary salivary gland epithelial cells (pSGECs) from rat submandibular (SM) gland and parotid gland (PG) was established and characterized. pSGECs grown on SFS, but not tissue culture plastic (TCP), formed aggregates of cells with morphological features resembling secretory acini. High levels of amylase were released into the media by both cell types after extended periods in culture on SFS. Remarkably, cultures of PG-derived cells on SFS, but not SM cells, responded to isoproterenol, a ?-adrenergic receptor agonist, with increased enzyme release. This behavior mimics that of the salivary glands in vivo. Decellularized extracellular matrix (ECM) formed by pSGECs in culture on SFS contained type IV collagen, a major component of the basement membrane. These results demonstrate that pSGECs grown on SFS, but not TCP, retain important functional and structural features of differentiated salivary glands and produce an ECM that mimics the native salivary gland cell niche. These results demonstrate that SFS has potential as a scaffold for creating the salivary gland cell niche in vitro and may provide an approach for inducing multipotent stem cells to provide therapeutically meaningful numbers of salivary gland progenitor cells for regenerating these tissues in patients. PMID:25625623
Production of raw-starch-digesting ?-amylase isoform from Bacillus sp. under solid-state fermentation and biochemical characterization.
Boži?, Nataša; Slavi?, Marinela Šokarda; Gavrilovi?, Anja; Vuj?i?, Zoran
?-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
Cloning Sequencing and Expression of the Gene Encoding Extracellular a-Amylase from Pyrococcus furiosus and Biochemical Characterization of the Recombinant Enzyme
Guoqiang Dong; Claire Vieille; Alexei Savchenko; J. Gregory Zeikus
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%
Digestive amylase from the larger grain borer, Prostephanus truncatus Horn
E Mendiola-Olaya; A Valencia-Jiménez; S Valdés-Rodr??guez; J Délano-Frier; A Blanco-Labra
A combination of ion-exchange chromatography, preparative electrophoresis and gel filtration chromatography allowed a 1209-fold purification of one of the two major digestive ?-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
Amlioration des plantes Polymorphisme de l'? amylase chez le pcher.
Paris-Sud XI, Université de
Amélioration des plantes Polymorphisme de l'? amylase chez le pêcher. Étude 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
Hyperglycaemia but not hyperlipidaemia decreases serum amylase and increases neutrophils in the exocrine pancreas of cats.
Zini, Eric; Osto, Melania; Moretti, Simona; Franchini, Marco; Kook, Peter H; Lutz, Hans; Guscetti, Franco; Perren, Aurel; Hoelzle, Ludwig E; Ackermann, Mathias; Lutz, Thomas A; Reusch, Claudia E
The goal of the study was to determine whether hyperglycaemia or hyperlipidaemia causes pancreatitis in cats and to assess the effect of excess serum glucose and lipids on amylase and lipase activity. Ten-day hyperglycaemic and hyperlipidaemic clamps were carried out in five and six healthy cats, respectively. Ten healthy cats received saline and served as controls. The activity of amylase was below the normal range in 4 of 5 hyperglycaemic cats by day 10. The activity of lipase did not vary in any of the cats. Samples of exocrine pancreas were normal on histological examination, but the number of tissue neutrophils was increased in hyperglycaemic cats (P<0.05). In a retrospective study 14 of 40 (35%) cats with naturally occurring diabetes mellitus had amylase activities below the reference range at the time of admission. Amylase activities normalised within 1 week of insulin therapy and subsequent glycaemic control. Lipase activity was increased in 26 of 40 (65%) diabetic cats and remained elevated despite glycaemic control. In conclusion, hyperglycaemia, but not hyperlipidaemia, increases pancreatic neutrophils in cats. However, because the histological morphology of the exocrine pancreas was normal, hyperglycaemia may play only a minor role in the pathogenesis of pancreatitis. Low amylase activities in diabetic cats may reflect an imbalance in glucose metabolism rather than pancreatitis. PMID:20132955
Effects of season, age, sex, and housing on salivary cortisol concentrations in horses.
Aurich, J; Wulf, M; Ille, N; Erber, R; von Lewinski, M; Palme, R; Aurich, C
Analysis of salivary cortisol is increasingly used to assess stress responses in horses. Because spontaneous or experimentally induced increases in cortisol concentrations are often relatively small for stress studies, proper controls are needed. This requires an understanding of the factors affecting salivary cortisol over longer times. In this study, we have analyzed salivary cortisol concentration for 6 mo in horses (n = 94) differing in age, sex, reproductive state, and housing. Salivary cortisol followed a diurnal rhythm with the highest concentrations in the morning and a decrease throughout the day (P < 0.001). This rhythm was disrupted in individual groups on individual days; however, alterations remained within the range of diurnal changes. Comparison between months showed highest cortisol concentrations in December (P < 0.001). Cortisol concentrations increased in breeding stallions during the breeding season (P < 0.001). No differences in salivary cortisol concentrations between nonpregnant mares with and without a corpus luteum existed. In stallions, mean daily salivary cortisol and plasma testosterone concentrations were weakly correlated (r = 0.251, P < 0.01). No differences in salivary cortisol between female and male young horses and no consistent differences between horses of different age existed. Group housing and individual stabling did not affect salivary cortisol. In conclusion, salivary cortisol concentrations in horses follow a diurnal rhythm and are increased in active breeding sires. Time of the day and reproductive state of the horses are thus important for experiments that include analysis of cortisol in saliva. PMID:25700267
Ultrastructural localization of salivary mucins MUC5B and MUC7 in human
Terasaki, Mark
mucosal surfaces with highly glycosylated proteins that are active in bacterial aggregation and in oral, in maintaining the integrity of oral structures, and in the maintenance of oral health. Human salivary glands of the oral mucosa (1). Although saliva is produced mostly by the three major salivary glands (parotid
Analysis of ?-amylase inhibitor from corni fructus by coupling magnetic cross-linked enzyme aggregates of ?-amylase with HPLC-MS.
Liu, Liangliang; Cen, Yin; Liu, Fang; Yu, Jingang; Jiang, Xinyu; Chen, Xiaoqing
As a carrier-free immobilization strategy, magnetic cross-linked enzyme aggregates (MCLEAs) showed improved enzyme activity, stability and magnetic response. In this study, MCLEAs of ?-amylase (MCLEAs-amylase) was prepared under optimized conditions and characterized with scanning electron microscope and vibrating sample magnetometer. The prepared MCLEAs-amylase showed an amorphous structure and the saturation magnetization was 33.5emu/g, which was sufficient for magnetic separation. Then MCLEAs-amylase coupled with high performance liquid chromatography-mass spectrometry (HPLC-MS) was utilized to screen and identify ?-amylase inhibitors from ethyl acetate extract of corni fructus. The experiment conditions were optimized. At the optimum conditions (incubation time: 10min, pH: 7.0 and temperature: 20°C), querciturone was successfully screened and identified with weak non-specific binding. The screening result was verified by inhibition assays and the IC50 value of querciturone was 22.5?g/mL. This method provided a rapid way to screen active compounds from natural products. PMID:26038235
Screening, Gene Cloning, and Characterizations of an Acid-Stable ?-Amylase.
Liu, Xinyu; Jia, Wei; An, Yi; Cheng, Kun; Wang, Mingdao; Yang, Sen; Chen, Hongge
Based on its ?-amylase activity at pH 5.0 and optimal pH of the crude enzyme, a strain (named B-5) with acid ?-amylase production was screened. The B-5 strain was identified as Bacillus amyloliquefaciens through morphological, physiological, and biochemical characteristics analysis, as well as 16S rDNA phylogenetic analysis. Its ?-amylase gene of GenBank Accession No. GU318401 was cloned and expressed in Escherichia coli. The purified recombinant ?-amylase AMY-Ba showed the optimal pH of 5.0, and was stable at a pH range of 4.0-6.0. When hydrolyzing soluble starch, amylose, and amylopectin, AMY-Ba released glucose and maltose as major end products. The ?-amylase AMY-Ba in this work was a different type from the well-investigated J01542 (GenBank Accession No.)-type ?-amylase from the same species. AMY-Ba exhibited notable adsorption and hydrolysis ability towards various raw starches. Structure analysis of AMY-Ba suggested the presence of a new starch-binding domain at its C-terminal region. PMID:25563420
Geographical polymorphism of amylase in Drosophila ananassae and its relatives.
Da Lage, J L; Cariou, M L; David, J R
Strains of Drosophila ananassae from various places in the Tropics were investigated for their electrophoretical amylase pattern. Eleven isoamylases were found in adult flies. African populations were much more polymorphic than those from the Far East, and showed multibanded phenotypes, suggesting a multiplication of the Amy structural gene, with at least four copies per haploid genome in certain populations. Nine other species of the D. ananassae subgroup had weak amylase activity and only one or two variants were found in each species. D. monieri and D. varians are closely related to D. ananassae and showed a single band, similar to the isoamylase 3 of D. ananassae, which suggests that this might be an ancestral allele. PMID:2475457
Serum Amylase in Bulimia Nervosa and Purging Disorder: Differentiating the Association with Binge Eating versus Purging Behavior
Wolfe, Barbara E.; Jimerson, David C.; Smith, Adrian; Keel, Pamela K.
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
Some chemical characteristics of human minor salivary gland secretions.
Hensten-Pettersen, A
The minor salivary glands contribute to the composition of whole saliva, but little information has been available about their chemical constituents. Pilocarpine-stimulated labial and palatine secretion from 4 human subjects was investigated by paper and disc electrophoresis, immunochemical analysis, and for content of carbohydrates, amino acids, lipids, hexuronic acids and sulphate. No significant differences were noted between the labial and palatine secretions by any of the methods employed. The minor gland secretions appeared to consist mainly of mucosubstances, possibly with blood group specificity. In addition, three water-soluble components with the characteristics of albumin, alpha-amylase and secretory IgA were seen. The minor gland secretions had an amino acid profile different from those of the major salivary glands and contained higher proportions of carbohydrate. Only one lipid component, with the characteristics of a polar lipid, was seen. Hexuronic acids were not detected in either secretion, whereas both contained sulphate. It would appear that the minor mucous glands contribute to the content of mucosubstances in whole saliva, whereas their content of water-soluble material is negligible in this respect. PMID:822684
Human Salivary Alpha-Amylase (EC.18.104.22.168) Activity and Periodic Acid and Schiff Reactive (PAS) Staining: A Useful Tool to Study Polysaccharides at an Undergraduate Level
ERIC Educational Resources Information Center
Fernandes, Ruben; Correia, Rossana; Fonte, Rosalia; Prudencio, Cristina
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…
Residual amylopectin structures of amylase-treated wheat starch slurries reflect amylase mode of action
Pedro Leman; Hans Goesaert; Jan A. Delcour
Some amylases can delay bread staling and\\/or starch (amylopectin) retrogradation, but the molecular basis of this effect remains little understood. In order to increase our insight in these aspects of amylase functionality, several amylases were added in a pure wheat-starch-containing model system and subjected to a heating step corresponding to that in the baking phase in bread making. Next, the
Stages of Salivary Gland Cancer
... treat some salivary gland tumors . These include: Fast neutron radiation therapy : Fast neutron radiation therapy is a type of high-energy ... radiation therapy machine aims tiny, invisible particles, called neutrons, at the cancer cells to kill them. Fast ...
Comparative analysis of the development of the mandibular salivary glands and the labial silk glands in the mulberry silkworm, Bombyx mori.
Parthasarathy, R; Gopinathan, Karumathil P
The mulberry silkworm, Bombyx mori has a pair of salivary glands arising from the mandibular segment, in addition to the labial silk glands which are generally considered as modified salivary glands. Here we report the characterization of salivary glands and the comparative gene expression profiling of the silk and salivary glands. The two independent salivary glands made up by 330 cells, grow about 1000 fold during larval development. These individual glands extend up to the T(1) thoracic segment unlike silk glands with fused anterior ends and extending up to the caudal region. The salivary glands also undergo endomitosis resembling the silk glands. The B. mori homologue of the homeotic gene Deformed (BmDfd) was expressed in the mandibular and maxillary segments in stage 17 embryo and got localized to the centre of the mandibular segment at stage 18 to form the salivary gland placodes. The expression was also seen in the distal ends of the leg appendages after blastokinesis (stage 22). Only low variations in BmDfd expression ranging from 1.6 to 2.1 fold were apparent during embryonic development. BmDfd expression was observed in the salivary glands all through the larval instars but not in the silk glands. The transcription factor, Forkhead and the segment polarity gene, Wingless were expressed throughout the salivary glands, the latter confirming the absence of physiological compartmentation within these glands unlike the silk glands. The expression of Amylase and Fibrohexamerin was restricted to the salivary and silk glands, respectively and therefore, served as molecular markers for these tissues. PMID:15661638
Self-compassion training modulates alpha-amylase, heart rate variability, and subjective responses to social evaluative threat in women.
Arch, Joanna J; Brown, Kirk Warren; Dean, Derek J; Landy, Lauren N; Brown, Kimberley D; Laudenslager, Mark L
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
Purification and properties of alpha-amylase from chicken (Gallus gallus L.) pancreas.
Buonocore, V; Deponte, R; Gramenzi, F; Petrucci, T; Poerio, E; Silano, V
Amylase from chicken pancreas was purified by an affinity method involving filtering a crude extract from pancreas through a Sepharose-wheat albumin column and eluting the retained enzyme with maltose. The purified amylase showed two active bands upon polyacrylamide electrophoresis in an alkaline buffer system and only one band in an acidic buffer system. The enzyme is a Ca2+-glycoprotein which behaves as a typical alpha-amylase. It consists of a single polypeptide chain with molecular weight 53,000 and contains 5.3 moles of reducing sugars per mole of protein. Optimal conditions of pH and temperature for the enzymic activity are 7.5 and 37 degrees C. The enzyme is irreversibly inactivated by removal of Ca2+ by exhaustive dialysis and is activated by the presence in the assay mixture of Cl-; other halides are less effective than Cl- in activating the enzyme. PMID:20568
Purification and Characterization of Midgut ?-Amylase in a Predatory Bug, Andralus spinidens
Sorkhabi-Abdolmaleki, Sahar; Zibaee, Arash; Hoda, Hassan; Fazeli-Dinan, Mahmoud
?-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
Longitudinal studies of luteal function by salivary progesterone determinations.
Walker, S M; Walker, R F; Riad-Fahmy, D
A 'normal range' for salivary progesterone concentrations has been established using data derived from women who were menstruating regularly and in whom dating of the cycle by accepted criteria was possible. Since these values are in agreement with those in the first 9 days of conception cycles and with those in cycles in which ovulation was confirmed by ultrasonography, they provide a reliable index of progesterone output compatible with fertility. Measurement of daily salivary progesterone values in subfertile women for time spans exceeding 3 months allowed accurate assessment of base-line ovarian activity and of the response to ovulation-induction therapy. Salivary sampling, by allowing collection of frequent samples with a minimum of time, stress and inconvenience, is ideally suited to longitudinal studies of ovarian activity. This sampling regimen is also applicable to the monitoring of progesterone output throughout pregnancy. PMID:6510896
Alkaline proteases and thermostable ?-amylase co-produced by Bacillus licheniformis NH1: Characterization and potential application as detergent additive
Noomen Hmidet; Nedra El-Hadj Ali; Anissa Haddar; Safia Kanoun; Sellami-Kamoun Alya; Moncef Nasri
Alkalophilic Bacillus licheniformis NH1 strain produced at least five major extracellular proteases and a unique amylase as showed by zymography technique. The optimum pH and temperature for the proteolytic activity were 10.0 and 70°C, respectively, while those of amylolytic activity were 6.5 and 90°C, respectively. The alkaline proteases and thermostable ?-amylase showed extreme stability towards non-ionic and anionic surfactants after
Isolation of a cDNA Clone for alpha-Amylase in Mung Bean Cotyledons : Analysis of alpha-Amylase mRNA Levels in Cotyledons during and following Germination of Mung Bean Seeds.
Koizuka, N; Tanaka, Y; Morohashi, Y
A cDNA was isolated that codes for alpha-amylase in mung bean (Vigna radiata) cotyledons, and the nucleotide sequence was determined. The deduced amino acid sequence (421 amino acid residues) is about 65% homologous with those of barley alpha-amylases. By comparing the deduced sequence with the sequence of the purified alpha-amylase, it was inferred that 23 N-terminal amino acids of a nascent polypeptide represent a signal peptide. Northern blot analysis showed that the levels of alpha-amylase mRNA are in parallel with the activities of alpha-amylase synthesis in cotyledons. Under the conditions where the solute leakage from cotyledons is accelerated during imbibition, a rapid increase in the amount of the alpha-amylase mRNA occurs. We postulate that a factor(s) which regulates in an inhibitory manner the alpha-amylase expression at the transcriptional level may be present in dry cotyledons and be removed by leakage. PMID:16667859
Random mutagenesis used to probe the structure and function of Bacillus stearothermophilus alpha-amylase.
Holm, L; Koivula, A K; Lehtovaara, P M; Hemminki, A; Knowles, J K
Mutations that cover the sequence of Bacillus stearothermophilus alpha-amylase were produced by an efficient in vitro enzymatic random mutagenesis method and the mutant alpha-amylases were expressed in Escherichia coli, which also secreted the product. Ninety-eight mutants were identified by sequencing and their enzyme activities were classified into three classes: wild-type, reduced or null. A molecular model of the enzyme was constructed using the coordinates of Takaamylase A and a consensus alignment of mammalian, plant, and bacterial alpha-amylases. The location of mutant amino acids on the model indicate that mutations which destroy or decrease the catalytic activity are particularly clustered: (i) around the active site and along the substrate-binding groove and (ii) in the interface between the central alpha/beta barrel and the C-terminal domain. Exposed loops are typically tolerant towards mutations. PMID:2330367
Conditional overexpression of TGF-?1 disrupts mouse salivary gland development and function
Hall, Bradford E.; Zheng, Changyu; Swaim, William D.; Cho, Andrew; Nagineni, Chandrasekharam N.; Eckhaus, Michael A.; Flanders, Kathleen C.; Ambudkar, Indu S.; Baum, Bruce J.; Kulkarni, Ashok B.
Transforming growth factor-? (TGF-?) signaling is known to affect salivary gland physiology by influencing branching morphogenesis, regulating ECM deposition, and controlling immune homeostasis. To study the role of TGF-?1 in the salivary gland, we created a transgenic mouse (?1glo) that conditionally over-expresses the active TGF-?1 upon genomic recombination by the Cre recombinase. The ?1glo mice were bred with a MMTV (mouse mammary tumor virus)-Cre (MC) transgenic line that expresses the Cre recombinase predominantly in the secretory cells of both the mammary and salivary glands. Although most of the double positive (?1glo/MC) pups die either in utero or just after birth, clear defects in salivary gland morphogenesis could be seen such as reduced branching and increased mesenchyme. The ?1glo/MC mice that survived into adulthood, however, had hyposalivation due to salivary gland fibrosis and acinar atrophy. Increased TGF-? signaling was observed in the salivary gland with elevated phosphorylation of Smad2 and a concomitant increase in ECM deposition. In particular, aberrant TGF-?1 overexpression caused salivary gland hypofunction in this mouse model because of the replacement of normal glandular parenchyma with interstitial fibrous tissue. These results further implicate TGF-? in pathological cases of salivary gland inflammation and fibrosis that occur with chronic infections in the glands or with the autoimmune disease, Sjögren's syndrome or with the radiation therapy given to head-and-neck cancer patients. PMID:20142803
Ca2+-dependent K+ Channels in Exocrine Salivary Glands
Catalán, Marcelo A.; Peña-Munzenmayer, Gaspar; Melvin, James E.
In the last 15 years, remarkable progress has been realized in identifying the genes that encode the ion-transporting proteins involved in exocrine gland function, including salivary glands. Among these proteins, Ca2+-dependent K+ channels take part in key functions including membrane potential regulation, fluid movement and K+ secretion in exocrine glands. Two K+ channels have been identified in exocrine salivary glands: 1) a Ca2+-activated K+ channel of intermediate single channel conductance encoded by the KCNN4 gene; and, 2) a voltage- and Ca2+-dependent K+ channel of large single channel conductance encoded by the KCNMA1 gene. This review focuses on the physiological roles of Ca2+-dependent K+ channels in exocrine salivary glands. We also discuss interesting recent findings on the regulation of Ca2+-dependent K+ channels by protein-protein interactions that may significantly impact exocrine gland physiology. PMID:24559652
[Salivary hormones: a new aspect of oral physiology].
Deville de Périère, D; Arancibia, S
Aside from the digestive enzymes the submandibular salivary glands (SSG) synthetize other polypeptides, detected also in saliva, with varied biological activity; NGF and EGF are the knowest. However, over the last decade, steroids hormones have been also found out in the saliva at the same concentrations that the free plasma fraction. The origin of these hormones is largely discussed and certain authors have even proposed a local synthesis for them. This matter, is of clinical interest because gingiva and buccal tissues are knowingly sensitive to steroids. Besides, woman ovulation appears to be monitored through progesterone fluctuations in saliva. Another kind of salivary substances is formed by the neuropeptides of the gut-brain axis, mainly VIP and SRIF. The former likely of nervous origin seems to be involved in the atropine-resistant salivary secretion, whereas the latter-likely of SSG origin--appears as a factor associated with glycemia control. PMID:3078705
Sensitivity of Salivary Glands to Radiation
Grundmann, O.; Mitchell, G.C.; Limesand, K.H.
Radiation therapy for head and neck cancer causes significant secondary side-effects in normal salivary glands, resulting in diminished quality of life for these individuals. Salivary glands are exquisitely sensitive to radiation and display acute and chronic responses to radiotherapy. This review will discuss clinical implications of radiosensitivity in normal salivary glands, compare animal models used to investigate radiation-induced salivary gland damage, address therapeutic advances, and project future directions in the field. PMID:19783796
Purification and characterization of novel ?-amylase from Bacillus subtilis KIBGE HAS.
Bano, Saeeda; Ul Qader, Shah Ali; Aman, Afsheen; Syed, Muhammad Noman; Azhar, Abid
Purification of extracellular ?-amylase from Bacillus subtilis KIBGE HAS was carried out by ultrafiltration, ammonium sulfate precipitation and gel filtration chromatography. The enzyme was purified to homogeneity with 96.3-fold purification with specific activity of 13011 U/mg. The molecular weight of purified ?-amylase was found to be 56,000 Da by SDS-PAGE. Characteristics of extracellular ?-amylase showed that the enzyme had a Km and V (max) value of 2.68 mg/ml and 1773 U/ml, respectively. The optimum activity was observed at pH 7.5 in 0.1 M phosphate buffer at 50 °C. The amino acid composition of the enzyme showed that the enzyme is rich in neutral/non polar amino acids and less in acidic/polar and basic amino acids. The N-terminal protein sequence of 10 residues was found to be as Ser-Ser-Asn-Lys-Leu-Thr-Thr-Ser-Trp-Gly (S-S-N-K-L-T-T-S-W-G). Furthermore, the protein was not N-terminally blocked. The sequence of ?-amylase from B. subtilis KIBGE HAS was a novel sequence and showed no homology to other reported ?-amylases from Bacillus strain. PMID:21234823
Enzymatic detergent formulation containing amylase from Aspergillus niger: a comparative study with commercial detergent formulations.
Mitidieri, Sydnei; Souza Martinelli, Anne Helene; Schrank, Augusto; Vainstein, Marilene Henning
There is a wide range of biotechnological applications for amylases, including the textile, pharmaceutical, food and laundry industries. Hydrolytic enzymes are 100% biodegradable and enzymatic detergents can achieve effective cleaning with lukewarm water. Microorganisms and culture media were tested for amylase production and the best producer was Aspergillus niger L119 (3.9 U ml(-1) +/- 0.2) in submerged culture and its amylase demonstrated excellent activity at 50-55 degrees C and pH 4.0, remaining stable at 53 degrees C for up to 200 h. In order to establish the potential uses of this enzyme in detergents, different formulations were tested using the A. niger amylase extract. Enzyme activity was compared with three commercial formulations. The detergents are used in hospitals to clean surgical and endoscopy equipment. The presence of amylase in the formulation is because of its action within hospital drainage system, whether or not it has any function in cleaning the equipment. PMID:16112858
Bioassay-guided separation of an alpha-amylase inhibitor anthocyanin from Vaccinium arctostaphylos berries.
Nickavar, Bahman; Amin, Gholamreza
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
Bioactivity-Guided Separation of an ?-Amylase Inhibitor Flavonoid from Salvia virgata
Nickavar, Bahman; Abolhasani, Leyla
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
Production and partial purification of alpha-amylase from a novel isolate Streptomyces gulbargensis.
Syed, Dastager G; Agasar, Dayanand; Pandey, Ashok
Extracellular amylase production by a newly isolated alkali-thermotolerant strain Streptomyces gulbargensis DAS 131 was optimized and characterized. The highest amylase production was achieved by growing S. gulbargensis DAS 131 in media with 1% starch. Strain exhibited maximal activity at pH 9.0 and 45 degrees C and relatively stable in alkaline conditions (pH 11). Starch and peptone were found to be the good source of carbon and nitrogen with a yield of 2,216.6 and 2,156.1 U, respectively. Maltose and maltotriose were the main end products of starch hydrolysis, indicating alpha-amylase activity. SDS-PAGE analysis revealed a monomeric form with a molecular weight of 55 kDa. PMID:18846397
Bioactivity-Guided Separation of an ?-Amylase Inhibitor Flavonoid from Salvia virgata.
Nickavar, Bahman; Abolhasani, Leyla
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
Effect of cadmium on germination, amylases and rate of respiration of germinating pea seeds
L. K. Chugh; S. K. Sawhney
Growth of embryonic axis of germinating pea seeds (Pisum sativum cv. Bonneville) was significantly inhibited by as low as 0.25 mM cadmium and the elongation of the radicle was affected more severely than that of the plumule. Total amylolytic activity, as well as activities of ?- and ?-amylases, diminished progressively with increasing concentrations of the metal in the media. The
Effects of psychosocial stress on the pattern of salivary protein release.
Trueba, Ana F; Mizrachi, Dario; Auchus, Richard J; Vogel, Pia D; Ritz, Thomas
Previous research suggests that acute stress can increase the release of immune-relevant proteins in saliva. However, no attempts have been made to examine a wider range of salivary proteins in response to stress. In this study, we identified and quantified changes in the pattern of salivary protein release in a 45 min time period following the Trier Social Stress Test (TSST) in 12 asthmatic and 13 healthy participants. Proteins were separated using sodium dodecyl sulfate polyacrylamide gel electrophoresis. The relative protein amounts were quantified using the Image J software (NIH), and identified and characterized using mass spectroscopy. Negative affect was increased immediately after stress in both groups. The results showed that alpha amylase, cystatin S and light chain IgA were increased after the TSST and significant increases in glutathione S-transferase and prolactin inducible protein were also observed. Asthma patients showed responses similar to healthy controls, but had a tendency toward overall lower alpha amylase levels. Our findings suggest that a variety of proteins relevant to mucosal immunity are elevated following acute psychosocial stress, including glutathione S-transferase and prolactin inducible protein, which had not been characterized in this context before. PMID:22056539
Expression of Adrenomedullin and its Receptors in Human Salivary Tissue
S. Kapas; K. Pahal; A. T. Cruchley; E. Hagi-Pavli; J. P. Hinson
Adrenomedullin is a multifunctional peptide produced by a wide range of different cells and tissues. This study was designed to investigate whether adrenomedullin is present in human saliva and in salivary glands. It was expected that saliva may contain high concentrations of adrenomedullin, which has antimicrobial activity in vitro, which may have functional implications in the oral cavity. Saliva from
Highly thermostable, thermophilic, alkaline, SDS and chelator resistant amylase from a thermophilic Bacillus sp. isolate A3-15
Burhan Arikan
A thermostable alkaline ?-amylase producing Bacillus sp. A3-15 was isolated from compost samples. There was a slight variation in amylase synthesis within the pH range 6.0 and 12.0 with an optimum pH of 8.5 (8mm zone diameter in agar medium) on starch agar medium. Analyses of the enzyme for molecular mass and amylolytic activity were carried out by starch SDS–PAGE
Halophilic alkali- and thermostable amylase from a novel polyextremophilic Amphibacillus sp. NM-Ra2.
Mesbah, Noha M; Wiegel, Juergen
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
Irradiation induced salivary gland neoplasia
Sener, S.F.; Scanlon, E.F.
Twenty-six patients with a prior history of irradiation for benign conditions of the head and neck and salivary gland abnormalities are reported. All the patients had preoperative physical findings suggestive of tumor, not glandular infection. Forty-six per cent of the patients had one carcinoma and 11% had two carcinomas within the irradiated field. Eight of the 11 malignant tumors in these 26 patients were in the parotid gland. The nonmalignant salivary changes were similar to those previously reported in glands receiving therapeutic irradiation for carcinoma.
?-Amylase from Starchless Seeds of Trigonella Foenum-Graecum and Its Localization in Germinating Seeds
Srivastava, Garima; Kayastha, Arvind M.
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
Production of fungal alpha-amylase by Saccharomyces kluyveri in glucose-limited cultivations.
Møller, Kasper; Sharif, Mostafa Z; Olsson, Lisbeth
Heterologous protein production by the yeast Saccharomyces kluyveri was investigated under aerobic glucose-limited conditions. Alpha-amylase from Aspergillus oryzae was used as model protein and the gene was expressed from a S. cerevisiae 2 micro plasmid. For comparison, strains of both S. kluyveri and S. cerevisiae were transformed with the same plasmid, which led to secretion of active alpha-amylase in both cases. The S. cerevisiae 2 micro plasmid was found to be stable in S. kluyveri as evaluated by a constant alpha-amylase productivity in a continuous cultivation for more than 40 generations. S. kluyveri and S. cerevisiae secreted alpha-amylase with similar yields during continuous cultivations at dilution rates of 0.1 and 0.2 h(-1) (4.8-5.7 mg (g dry weight)(-1)). At a dilution rate of 0.3 h(-1) the metabolism of S. kluyveri was fully respiratory, whereas S. cerevisiae produced significant amounts of ethanol. A fed-batch cultivation was carried out with S. kluyveri where the biomass concentration reached 85 g l(-1) and the alpha-amylase concentration reached 320 mg l(-1). Even though S. kluyveri could be grown to high cell density, it was also observed that it has a high maintenance coefficient, which resulted in low biomass yields at the low specific growth rates prevailing towards the end of the fed-batch cultivation. PMID:15246667
Substantivity of a single chlorhexidine mouthwash on salivary flora: Influence of intrinsic and extrinsic factors
I. Tomás; M. C. Cousido; L. García-Caballero; S. Rubido; J. Limeres; P. Diz
ObjectivesTo analyse the influence of intrinsic and extrinsic factors on the in vivo antimicrobial activity of a chlorhexidine (CHX) digluconate mouthwash on the salivary flora up to 7h after its application, using epifluorescence microscopy.
Experimental salivary pellicles formed on titanium surfaces mediate adhesion of streptococci.
Edgerton, M; Lo, S E; Scannapieco, F A
The goal of this study was to characterize salivary components of titanium pellicles and to determine how experimental pellicles affect adhesion of several strains of streptococci to titanium surfaces. Titanium experimental pellicles were formed by incubation of fresh human parotid or human submandibular-sublingual saliva on pure titanium beads. Pellicle was recovered from the beads using sodium dodecyl sulfate buffer and was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting to identify adsorbed salivary components. Streptococcus anginosus, S. oralis, and S. salivarius recovered from in vivo titanium plaque and five reference strains of streptococci were used in adhesion assays to titanium beads with and without experimental salivary pellicles. The experimental pellicle formed on titanium was found to be composed of selected proteins from human parotid and human submandibular-sublingual saliva. Salivary alpha-amylase and proline-rich proteins were found in all experimental pellicles, while sIgA, high-molecular weight mucin, and proline-rich glycoproteins were detected in one of the experimental pellicles examined. Adhesion of fresh isolates and reference stains of S. anginosus, S. oralis, and S. salivarius to saliva-coated titanium was reduced compared to that of titanium without saliva coating. However, adhesion of laboratory strains of S. gordonii and S. sanguis was found to be significantly greater to experimental pellicles of human submandibular-sublingual saliva than was the adhesion of the fresh isolates, suggesting that streptococci-colonizing implant surfaces may be inherently less adhesive than other bacterial strains. This study found that salivary pellicles are selectively formed on titanium and mediate in vitro adhesion of streptococci. PMID:8803339
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...
The Renal Handling of Amylase in Normal Man
K. Søiling; C. E. Mogensen; E. Vittinghus; A. Brock
The renal glomerular and tubular transport rate of amylase was studied by measuring the urinary excretion of this protein before and during inhibition of tubular protein reabsorption by lysine. The excretion of amylase was compared with the excretion of albumin, ?-2-microglobulin and free light chains of immunoglobulins. This investigation showed that amylase is reabsorbed by the tubular cells, but only
Pleomorphic adenoma of the buccal salivary gland
Khandekar, Shubhangi; Dive, Alka; Munde, Prashant; Wankhede, Neena Dongre
Salivary gland swellings can result from tumors, an inflammatory process or cysts. It can sometimes be difficult to establish; whether pathology arises from the salivary gland itself or adjacent structures. Neoplasms of the salivary glands account for less than 1% of all tumors, 3–5% of all head and neck tumors and benign pleomorphic adenoma (PA) of minor salivary glands arising de novo is very rare. PA is the most common tumor of the salivary gland. While the majority arises from the parotid gland, only a small percentage arises from the buccal minor salivary gland. A case of PA of minor salivary glands in the buccal mucosa in a 70-year-old female is discussed. It includes review of literature, clinical features, histopathology, radiological findings and treatment of the tumor; with emphasis on diagnosis.
Multiple amylase genes in Drosophila ananassae and related species.
Da Lage, J L; Lemeunier, F; Cariou, M L; David, J R
The number and organization of amylase genes in Drosophila ananassae were investigated through classical genetic methods and in situ and filter hybridizations. At least four genes may be active in D. ananassae, organized as two independent pairs of closely linked copies on the 2L and 3L chromosomal arms. Several other species of the D. ananassae subgroup were studied and show the same chromosomal locations, suggesting an ancient duplication event. However, the number of Amy copies seems to be higher in the D. ananassae multigene family, and there is a striking intraspecific molecular differentiation. PMID:1378417
Modulation of Sodium/Iodide Symporter Expression in the Salivary Gland
La Perle, Krista M.D.; Kim, Dong Chul; Hall, Nathan C.; Bobbey, Adam; Shen, Daniel H.; Nagy, Rebecca S.; Wakely, Paul E.; Lehman, Amy; Jarjoura, David
Background Physiologic iodide-uptake, mediated by the sodium/iodide symporter (NIS), in the salivary gland confers its susceptibility to radioactive iodine–induced damage following 131I treatment of thyroid cancer. Subsequent quality of life for thyroid cancer survivors can be decreased due to recurrent sialoadenitis and persistent xerostomia. NIS expression at the three principal salivary duct components in various pathological conditions was examined to better our understanding of NIS modulation in the salivary gland. Methods NIS expression was evaluated by immunohistochemistry in human salivary gland tissue microarrays constructed of normal, inflamed, and neoplastic salivary tissue cores. Cumulative 123I radioactivity reflecting the combination of NIS activity with clearance of saliva secretion in submandibular and parotid salivary glands was evaluated by single-photon emission computed tomography/computed tomography imaging 24 hours after 123I administration in 50 thyroid cancer patients. Results NIS is highly expressed in the basolateral membranes of the majority of striated ducts, yet weakly expressed in few intercalated and excretory duct cells. The ratio of 123I accumulation between parotid and submandibular glands is 2.38±0.19. However, the corresponding ratio of 123I accumulation normalized by volume of interest is 1.19±0.06. The percentage of NIS-positive striated duct cells in submandibular salivary glands was statistically greater than in parotid salivary glands, suggesting a higher clearance rate of saliva secretion in submandibular salivary glands. NIS expression in striated ducts was heterogeneously decreased or absent in sialoadenitis. Most ductal salivary gland tumors did not express NIS. However, Warthin's tumors of striated duct origin exhibited consistent and intense NIS staining, corresponding with radioactive iodine uptake. Conclusions NIS expression is tightly modulated during the transition of intercalated to striated ducts and striated to excretory ducts in salivary ductal cells. NIS expression in salivary glands is decreased during inflammation and tumor formation. Further investigation may identify molecular targets and/or pharmacologic agents that allow selective inhibition of NIS expression/activity in salivary glands during radioactive iodine treatment. PMID:23441638
Trans-chalcone: a novel small molecule inhibitor of mammalian alpha-amylase.
Najafian, Mahmoud; Ebrahim-Habibi, Azadeh; Hezareh, Nastaran; Yaghmaei, Parichehreh; Parivar, Kazem; Larijani, Bagher
Trans-chalcone (1,3-diphenyl-2-propen-1-one), a biphenolic core structure of flavonoids precursor was tested for inhibitory activity toward alpha-amylase. Porcine pancreatic alpha-amylase was observed to be effectively inhibited by this compound, which showed competitive behavior with a K(i) of 48 ?M. Soluble starch (the natural substrate of the enzyme) was used in this study in order to obtain more realistic results. The possible binding mode of the compound was assessed in silico, and the two residues Trp59, and Tyr62 were proposed as main interacting residues with trans-chalcone. In conclusion, this compound could be used to design effective inhibitors of alpha-amylase. PMID:20857221
Kinetic study of the thermal denaturation of a hyperthermostable extracellular ?-amylase from Pyrococcus furiosus.
Brown, I; Dafforn, T R; Fryer, P J; Cox, P W
Hyperthermophilic enzymes are of industrial importance and interest, especially due to their denaturation kinetics at commercial sterilisation temperatures inside safety indicating time-temperature integrators (TTIs). The thermal stability and irreversible thermal inactivation of native extracellular Pyrococcus furiosus ?-amylase were investigated using differential scanning calorimetry, circular dichroism and Fourier transform infrared spectroscopy. Denaturation of the amylase was irreversible above a Tm of approximately 106°C and could be described by a one-step irreversible model. The activation energy at 121°C was found to be 316kJ/mol. Using CD and FT-IR spectroscopy it was shown that folding and stability greatly increase with temperature. Under an isothermal holding temperature of 121°C, the structure of the PFA changes during denaturation from an ?-helical structure, through a ?-sheet structure to an aggregated protein. Such data reinforces the use of P. furiosus ?-amylase as a labile species in TTIs. PMID:24063888
Insulin-Like Growth Factor-1 Preserves Salivary Gland Function After Fractionated Radiation
Limesand, Kirsten H., E-mail: email@example.com [Department of Physiological Sciences, University of Arizona, Tucson, AZ (United States); Department of Nutritional Sciences, University of Arizona, Tucson, AZ (United States); Avila, Jennifer L. [Department of Physiological Sciences, University of Arizona, Tucson, AZ (United States); Victory, Kerton; Chang, Hui-Hua; Shin, Yoon Joo; Grundmann, Oliver [Department of Nutritional Sciences, University of Arizona, Tucson, AZ (United States); Klein, Rob R. [Department of Pathology, University of Arizona, Tucson, AZ (United States)
Purpose: Radiotherapy for head-and-neck cancer consists of fractionated radiation treatments that cause significant damage to salivary glands leading to chronic salivary gland dysfunction with only limited prevention and treatment options currently available. This study examines the feasibility of IGF-1 in preserving salivary gland function following a fractionated radiation treatment regimen in a pre-clinical model. Methods and Materials: Mice were exposed to fractionated radiation, and salivary gland function and histological analyses of structure, apoptosis, and proliferation were evaluated. Results: In this study, we report that treatment with fractionated doses of radiation results in a significant level of apoptotic cells in FVB mice after each fraction, which is significantly decreased in transgenic mice expressing a constitutively active mutant of Akt1 (myr-Akt1). Salivary gland function is significantly reduced in FVB mice exposed to fractionated radiation; however, myr-Akt1 transgenic mice maintain salivary function under the same treatment conditions. Injection into FVB mice of recombinant insulin-like growth factor-1 (IGF-1), which activates endogenous Akt, suppressed acute apoptosis and preserved salivary gland function after fractionated doses of radiation 30 to 90 days after treatment. FVB mice exposed to fractionated radiation had significantly lower levels of proliferating cell nuclear antigen-positive salivary acinar cells 90 days after treatment, which correlated with a chronic loss of function. In contrast, FVB mice injected with IGF-1 before each radiation treatment exhibited acinar cell proliferation rates similar to those of untreated controls. Conclusion: These studies suggest that activation of IGF-1-mediated pathways before head-and-neck radiation could modulate radiation-induced salivary gland dysfunction and maintain glandular homeostasis.
Local entropy difference upon a substrate binding of a psychrophilic ?-amylase and a mesophilic homologue
NASA Astrophysics Data System (ADS)
Kosugi, Takahiro; Hayashi, Shigehiko
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.
Trans -chalcone: a novel small molecule inhibitor of mammalian alpha-amylase
Mahmoud Najafian; Azadeh Ebrahim-Habibi; Nastaran Hezareh; Parichehreh Yaghmaei; Kazem Parivar; Bagher Larijani
Trans-chalcone (1,3-diphenyl-2-propen-1-one), a biphenolic core structure of flavonoids precursor was tested for inhibitory activity\\u000a toward alpha-amylase. Porcine pancreatic alpha-amylase was observed to be effectively inhibited by this compound, which showed\\u000a competitive behavior with a K\\u000a \\u000a i\\u000a of 48 ?M. Soluble starch (the natural substrate of the enzyme) was used in this study in order to obtain more realistic results.\\u000a The possible
[Optimization of cultivation conditions of the alpha-amylase producer Bacillus subtilis 147].
Avdiiuk, K V; Varbanets', L D
The influence of some technological parameters of cultivation of the producer Bacillus subtilis 147 on the process of extracellular enzymes alpha-amylase synthesis was investigated. The optimum sources of carbon (0.1% insoluble starch) and nitrogen (0.2% sodium nitrate) for maximum production of alpha-amylase are established. It was shown, that the temperature 42 degrees C, the carbon nitrogen ratio 1:2, pH 7.0, volume of a nutritious medium 100 ml, rotation rate 220 rev/min during 4-6 days are optimum parameters of the producer cultivation. As a result the enzyme activity was increased 14 times. PMID:18416149
Enhanced extracellular production of ?-amylase in Bacillus subtilis by optimization of regulatory elements and over-expression of PrsA lipoprotein.
Chen, Jingqi; Gai, Yuanming; Fu, Gang; Zhou, Wenjuan; Zhang, Dawei; Wen, Jianping
?-Amylase was used as a heterologous model protein to investigate the effects of promoters, signal peptides and over-expression of an extra-cytoplasmic molecular chaperone, PrsA lipoprotein, on enhancing the secretion of ?-amylase in Bacillus subtilis. Four promoters and six signal peptides were compared, successively, and the highest yield of ?-amylase was achieved under the promotion mediated by PAprE, a strong constitutive promoter, and secretion by SPnprE, a signal peptide from B. subtilis. Moreover, under conditions of overexpressed PrsA lipoprotein, the secretion production and activity of ?-amylase increased to 2.5-fold. The performance of the recombinant B. subtilis 1A751PL31 was evaluated with a fed-batch fermentation in a 7.5 l fermentor. Optimization of regulatory elements and over-expression of PrsA lipoprotein had a significant effect on enhancing the production of ?-amylase in B. subtilis. PMID:25515799
Isolation and characterization of a proteinaceous ?-amylase inhibitor AAI-CC5 from Streptomyces sp. CC5, and its gene cloning and expression.
Sun, Zhibin; Lu, Weihao; Liu, Pingping; Wang, Hui; Huang, Yan; Zhao, Yuguo; Kong, Yi; Cui, Zhongli
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
Study of fluorescence quenching of Barley ?-amylase
NASA Astrophysics Data System (ADS)
Bakkialakshmi, S.; Shanthi, B.; Bhuvanapriya, T.
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.
Adsorption of ?-amylase onto poly(acrylamide/maleic acid) hydrogels
NASA Astrophysics Data System (ADS)
Tümtürk, H.; Çaykara, T.; ?en, M.; Güven, O.
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.
Identification and analysis of the amylase-binding protein B (AbpB) and gene (abpB) from Streptococcus gordonii.
Li, Lina; Tanzer, Jason M; Scannapieco, Frank A
The binding of salivary amylase to Streptococcus gordonii has previously been shown to involve a 20-kDa amylase-binding protein (AbpA). S. gordonii also releases an 82-kDa protein into the supernatant that binds amylase. To study this 82-kDa component, proteins were precipitated from bacterial culture supernatants by the addition of acetone or purified amylase. Precipitated proteins were separated by SDS-PAGE and transferred to a sequencing membrane. The P2 kDa band was then sequenced, yielding a 25 N-terminal amino acid sequence, CGFIFGRQLTADGSTMFGPTEDYP. Primers derived from this sequence were used in an inverse PCR strategy to clone the full-length gene from S. gordonii chromosomal DNA. An open reading frame of 1959 bp was noted that encoded a 652 amino acid protein having a predicted molecular mass of 80 kDa. The first 24 amino acid residues were consistent with a hydrophobic signal peptide, followed by a 25 amino acid N-terminal sequence that shared identity (24 of 25 residues) with the amino acid sequence of purified AbpB. The abpB gene from strains of S. gordonii was interrupted by allelic exchange with a 420-bp fragment of the abpB gene linked to an erythromycin cassette. The 82-kDa protein was not detected in supernatants from these mutants. These abpB mutants retained the ability to bind soluble amylase. Thus, AbpA, but not AbpB, appears sufficient to be the major receptor for amylase binding to the streptococcal surface. The role of AbpB in bacterial colonization remains to be elucidated. PMID:12113927
Relationship of sequence and structure to specificity in the ?-amylase family of enzymes
E. Ann MacGregor; Štefan Jane?ek; Birte Svensson
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
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...
Inhibitory effect of hexane fraction from Myagropsis myagroides on pancreatic ?-amylase in vitro.
Pak, Won-Min; Kim, Koth-Bong-Woo Ri; Kim, Min-Ji; Cho, Ji-Young; Ahn, Dong-Hyun
A Myagropsis myagroides (Mm) methanol extract showed ?-amylase inhibitory activity of 13% at a concentration of 5 mg/ml. Results showed that the hexane fraction from the Mm methanol extract exhibited ?-amylase inhibitory activity with an IC50 value of 4.24 mg/ml. The hexane fraction was separated using silica-gel column chromatography, and six subfractions were obtained. The fraction eluted with CHCl3:MeOH = 50:1 showed the highest ?-amylase inhibitory activity with an IC50 value of 0.72 mg/ml. This fraction was purified using Sephadex LH-20 column chromatography and an octadecyl silica (ODS) Sepak cartridge, obtaining seven subfractions. Fraction (Fr.) 4 also showed a strong ?-amylase inhibitory activity with an IC50 value of 0.75 mg/ml. Fr. 4 was purified by Sephadex LH-20 column chromatography and ODS Sepak cartridge, obtaining six subfractions. Fr. 4-2 was identified as sargachromanol I with an IC50 value of 0.40 mg/ml, and the inhibition pattern analyzed from Lineweaver-Burk plots revealed it to be an uncompetitive inhibitor. These results suggest that Mm has potential as a natural antidiabetes agent. PMID:25315050
Modes of Inhibition of ?-Amylase and ?-Glucosidase by Aqueous Extract of Morinda lucida Benth Leaf
Kazeem, M. I.; Adamson, J. O.; Ogunwande, I. A.
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
Anatomy, biogenesis, and regeneration of salivary glands
Holmberg, Kyle V.; Hoffman, Matthew P.
An overview of the anatomy and biogenesis of salivary glands is important in order to understand the physiology, functions and disorders associated with saliva. A major disorder of salivary glands is salivary hypofunction and resulting xerostomia, or dry mouth, which affects hundreds of thousands of patients per year who suffer from salivary gland diseases or undergo head and neck cancer treatment. There is currently no curative therapy for these patients. To improve these patients' quality of life, new therapies are being developed based on findings in salivary gland cell and developmental biology. Here we discuss the anatomy and biogenesis of the major human salivary glands and the rodent submandibular gland (SMG), which has been used extensively as a research model. We also include a review of recent research on the identification and function of stem cells in salivary glands, and the emerging field of research suggesting nerves play an instructive role during development and may be essential for adult gland repair and regeneration. Understanding the molecular mechanisms involved in gland biogenesis provides a template for regenerating, repairing or reengineering diseased or damaged adult human salivary glands. We provide an overview of three general approaches currently being developed to regenerate damaged salivary tissue, including gene therapy, stem cell-based therapy, and tissue engineering. In the future, it may be that a combination of all three will be used to repair, regenerate and reengineer functional salivary glands in patients to increase the secretion of their saliva, the focus of this monograph. PMID:24862590
Determination of arginine catabolism by salivary pellet?
Hoogenkamp, M.A.; ten Cate, J.M.
To determine the formation of ammonium from arginine by oral bacteria residing in saliva and dental plaque, an arginolytic activity assay based on the work described by Nascimento et al. [2] was developed. Following the original methodology, insufficient ammonium production could be determined. To improve the method for our research goal, the following modifications were made to the original protocols:•The following changes were made to the arginine catabolism assay resulting in a 1000-fold increase in sensitivity: (i) the salivary pellet was washed and concentrated five times resulting in the removal of low density compounds interfering with the assay, (ii) the pH of the Tris–maleate buffer was increased from 6.0 to 7.5 resulting in a better conversion of arginine to ammonium and (iii) the incubation time was increased to 3 h to ensure that non-responders and salivary pellets low in cell numbers could yield detectable levels of ammonium.•Removal of a centrifuge step from the protein determination resulted in a higher protein yield improving the accuracy of the assay.•Changing from the use of the toxic, environmentally hazardous, mercury containing Nessler's reagent to a colorimetric enzyme assay achieved a safer and greener determination of ammonium concentration.
Spectral effect: each population must have its own normal midnight salivary cortisol reference values determined
Tanakol, Refik; Karpuzoglu, Hande; Abbasoglu, Semra; Yarman, Sema; Boztepe, Harika; Alagol, Faruk
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
Direct radioimmunoassay (RIA) of salivary testosterone: correlation with free and total serium testosterone
Vittek, J.; L'Hommedieu, D.G.; Gordon, G.G.; Rappaport, S.C.; Southren, A.L.
Simple and sensitive direct RIA for determination of salivary testosterone was developed by using RSL NOSOLVEX TM (125 1) kit produced by Radioassay System Laboratories (Carcon, California). In addition, a relationship between salivary and serum free and total testosterone concentrations was studied in randomly selected 45 healthy subjects, 5 females on oral contraceptive pills and 28 hypertensive patients on various treatment regimens. The lowest weight of testosterone detectable by the modified method was equivalent to 1 pg/ml of saliva, taking into account analytical variability. Intra- and interassay coefficients of variation were 5.09 +/- 2.7% and 8.2 +/- 5.9% respectively. Statistically significant correlations were found between salivary and serum free testosterone (r = 0.97) and salivary and serum total testosterone concentrations (r = 0.70 - 0.87). The exception to this was a group of hypertensive females in which no correlation (r = 0.14) between salivary and total serum testosterone was found. It is also of interest that, while salivary testosterone was significantly increased in subjects taking oral contraceptives and most of the hypertensive patients, the total serum testosterone concentration was in normal range. These findings suggest that the determination of salivary testosterone is a reliable method to detect changes in the concentration of available biologically active hormone in the circulation. 21 references, 4 figures, 1 table.
Genetic Regulation of Tissue-Specific Expression of Amylase Structural Genes in Drosophila melanogaster
Irene Abraham; Winifred W. Doane
Laboratory strains of Drosophila melanogaster were screened for spatial variations in adult midgut alpha -amylase (1,4-alpha -D-glucan glucanohydrolase, EC 184.108.40.206) 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.
Tobacco Plants Transformed with the Bean alphaai Gene Express an Inhibitor of Insect alpha-Amylase in Their Seeds.
Altabella, T; Chrispeels, M J
Bean (Phaseolus vulgaris L.) seeds contain a putative plant defense protein that inhibits insect and mammalian but not plant alpha-amylases. We recently (J Moreno, MJ Chrispeels [1989] Proc Natl Acad Sci USA 86:7885-7889) presented strong circumstantial evidence that this alpha-amylase inhibitor (alphaAI) is encoded by an already-identified lectin gene whose product is referred to as lectin-like-protein (LLP). We have now made a chimeric gene consisting of the coding sequence of the lectin gene that encodes LLP and the 5' and 3' flanking sequences of the lectin gene that encodes phytohemagglutinin-L. When this chimeric gene was expressed in transgenic tobacco (Nicotiana tabacum), we observed in the seeds a series of polypeptides (M(r) 10,000-18,000) that cross-react with antibodies to the bean alpha-amylase inhibitor. Most of these polypeptides bind to a pig pancreas alpha-amylase affinity column. An extract of the seeds of the transformed tobacco plants inhibits pig pancreas alpha-amylase activity as well as the alpha-amylase present in the midgut of Tenebrio molitor. We suggest that introduction of this lectin gene (to be called alphaai) into other leguminous plants may be a strategy to protect the seeds from the seed-eating larvae of Coleoptera. PMID:16667540
What Are the Key Statistics about Salivary Gland Cancer?
... 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 ...
Analysis of Candida albicans adhesion to salivary mucin.
Hoffman, M P; Haidaris, C G
Clearance of Candida albicans from the oral cavity is thought to be mediated via specific receptor-ligand interactions between salivary constituents and the fungus. Since surfaces in the oral cavity are normally coated with a saliva-derived pellicle, specific interactions between salivary constituents and C. albicans may also contribute to adhesion of C. albicans to the oral mucosa and dental prostheses. Therefore, the purpose of this study was to identify salivary constituents to which C. albicans is capable of binding. A solid-phase overlay assay was used in which electrophoretically separated rat and human salivary constituents bound to membrane filters were incubated with radiolabelled C. albicans cells. C. albicans adhered to a single salivary component from each host. Correlation of cell-binding activity with specific monoclonal antibody (MAb)-binding activity indicated that the constituent bound by C. albicans in human saliva was low-molecular-weight mucin (MG2) and that in rat saliva was rat submandibular gland (RSMG) mucin. Further studies showed an identical cell hybridization signal and MAb colocalization by using RSMG ductal saliva and an aqueous RSMG extract in the solid-phase overlay assay. Analysis of cell binding to the aqueous extract of RSMG fractionated by anion-exchange chromatography demonstrated that C. albicans binding was restricted to an acidic subfraction of the RSMG extract, which also bound the RSMG mucin-specific MAb. The Candida-binding fraction contained predominantly RSMG mucin glycoprotein and also a noncovalently associated, chloroform-extractable material. Furthermore, we identified two strains of C. albicans which differed severalfold in the ability to bind RSMG mucin in the overlay assay. These results suggest that C. albicans binds to only a specific subfraction of RSMG mucin and that the two C. albicans strains tested differ in the ability to bind RSMG mucin subfractions. Images PMID:8478083
Gender determines cortisol and alpha-amylase responses to acute physical and psychosocial stress in patients with borderline personality disorder.
Inoue, Ayako; Oshita, Harumi; Maruyama, Yoshihiro; Tanaka, Yoshihiro; Ishitobi, Yoshinobu; Kawano, Aimi; Ikeda, Rie; Ando, Tomoko; Aizawa, Saeko; Masuda, Koji; Higuma, Haruka; Kanehisa, Masayuki; Ninomiya, Taiga; Akiyoshi, Jotaro
Borderline personality disorder (BPD) is characterized by affective instability, unstable relationships, and identity disturbance. We measured salivary alpha-amylase (sAA) and salivary cortisol levels in all participants during exposure to the Trier Social Stress Test (TSST) and an electric stimulation stress. Seventy-two BPD patients were compared with 377 age- and gender- matched controls. The State and Trait versions of the Spielberger Anxiety Inventory test (STAI-S and STAI-T, respectively), the Profile of Mood State (POMS) tests, and the Beck Depression Inventory (BDI), the Depression and Anxiety Cognition Scale (DACS) were administered to participants before electrical stimulation. Following TSST exposure, salivary cortisol levels significantly decreased in female patients and significantly increased in male patients compared with controls. POMS tension-anxiety, depression-dejection, anger-hostility, fatigue, and confusion scores were significantly increased in BPD patients compared with controls. In contrast, vigor scores were significantly decreased in BPD patients relative to controls. Furthermore, STAI-T and STAI-S anxiety scores and BDI scores were significantly increased in BPD patient compared with controls. DACS scores were significantly increased in BPD patient compared with controls. Different stressors (e.g., psychological or physical) induced different responses in the HPA and SAM systems in female or male BPD patients. PMID:25979467
Tactics for modeling multiple salivary analyte data in relation to behavior problems: Additive, ratio, and interaction effects.
Chen, Frances R; Raine, Adrian; Granger, Douglas A
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
Purification, Characterization, and Potential of Saline Waste Water Remediation of a Polyextremophilic ?-Amylase from an Obligate Halophilic Aspergillus gracilis
Ali, Imran; Akbar, Ali; Yanwisetpakdee, Benjawan; Prasongsuk, Sehanat; Lotrakul, Pongtharin; Punnapayak, Hunsa
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
Larval salivary gland secretion proteins in Drosophila. Identification and characterization of the Sgs-5 structural gene.
Guild, G M; Shore, E M
The 90BC locus on the polytene chromosomal map of Drosophila melanogaster contains the structural gene for a third-instar, salivary gland-specific, polyadenylated RNA (the group V RNA). This also belongs to the intermolt puff set whose dispersed and co-ordinately regulated members are (1) transcriptionally active in the salivary gland during the third-instar developmental stage and (2) comprise (at least in part) the structural genes for a set of salivary gland secretion proteins. Previous developmental studies of the group V intermolt gene (located cytogenetically within the 90B3-8 interval) suggest that it controls the expression of a salivary gland secretion protein. By analyzing different D. melanogaster laboratory stocks for variation in group V gene expression, we have been able to correlate the presence of the group V RNA with the salivary gland secretion protein P4. In vitro translation experiments show that the salivary gland messenger RNA population derived from a stock that fails to synthesize the group V RNA does not direct the synthesis of a polypeptide similar in molecular weight to protein P4. In addition, cloned genomic DNA segments complementary to the group V RNA are capable of arresting the in vitro translation of this protein. Comparative two-dimensional fractionation of cysteine-labeled, protease-generated peptides shows that (1) the in vitro translation product arrested by group V gene DNA is biochemically very similar to or identical with the salivary gland secretion protein P4, and (2) protein P4 is equivalent to the salivary gland secretion protein previously designated SGS-5. Since designations of the latter type have been employed in naming the genetic loci that represent the structural genes for the salivary gland secretion protein gene set, the group V gene (previous designation) represents the SGS-5 structural gene and its appropriate genetic designation should now be Sgs-5. PMID:6439875
Effect of negative air ions on computer operation, anxiety and salivary chromogranin Alike immunoreactivity
Hideo Nakane; Osamu Asami; Yukio Yamada; Hideki Ohira
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
Sialolithiasis: an unusually large submandibular salivary stone.
Siddiqui, S J
Salivary gland calculi account for the most common disease of the salivary glands. The majority of sialoliths occur in the submandibular gland or its duct and are a common cause of acute and chronic infections. This case report describes a patient presenting with an unusually large submandibular gland sialolith, the subsequent patient management, the aetiology, diagnosis and various treatment modalities available for management of salivary gland calculi depending on their site and size. PMID:12199129
Salivary diagnostics: a brief review.
Malathi, Narasimhan; Mythili, Sabesan; Vasanthi, Hannah R
Early detection of disease plays a crucial role for treatment planning and prognosis. Saliva has great potential as a diagnostic fluid and offers advantage over serum and other biological fluids by an economic and noninvasive collection method for monitoring of systemic health and disease progression. The plethora of components in this fluid can act as biomarkers for diagnosis of various systemic and local diseases. In this review paper, we have emphasized the role of salivary biomarkers as diagnostic tools. PMID:24616813
Purification and characterization of a highly efficient calcium-independent ?-amylase from Talaromyces pinophilus 1-95.
Xian, Liang; Wang, Fei; Luo, Xiang; Feng, Yu-Liang; Feng, Jia-Xun
Alpha-amylase is a very important enzyme in the starch conversion process. Most of the ?-amylases are calcium-dependent and exhibit poor performance in the simultaneous saccharification and fermentation process of industrial bioethanol production that uses starch as feedstock. In this study, an extracellular amylolytic enzyme was purified from the culture broth of newly isolated Talaromyces pinophilus strain 1-95. The purified amylolytic enzyme, with an apparent molecular weight of 58 kDa on SDS-PAGE, hydrolyzed maltopentaose, maltohexaose, and maltoheptaose into mainly maltose and maltotriose and minor amount of glucose, confirming the endo-acting mode of the enzyme, and hence, was named Talaromyces pinophilus ?-amylase (TpAA). TpAA was most active at pH 4.0-5.0 (with the temperature held at 37°C) and 55°C (at pH 5.0), and stable within the pH range of 5.0-9.5 (at 4°C) and below 45°C (at pH 5.0). Interestingly, the Ca2+ did not improve its enzymatic activity, optimal temperature, or thermostability of the enzyme, indicating that the TpAA was Ca2+-independent. TpAA displayed higher enzyme activity toward malto-oligosaccharides and dextrin than other previously reported ?-amylases. This highly active Ca2+-independent ?-amylase may have potential applications in starch-to-ethanol conversion process. PMID:25811759
Crystal structures of the psychrophilic alpha-amylase from Alteromonas haloplanctis in its native form and complexed with an inhibitor.
Aghajari, N.; Feller, G.; Gerday, C.; Haser, R.
Alteromonas haloplanctis is a bacterium that flourishes in Antarctic sea-water and it is considered as an extreme psychrophile. We have determined the crystal structures of the alpha-amylase (AHA) secreted by this bacterium, in its native state to 2.0 angstroms resolution as well as in complex with Tris to 1.85 angstroms resolution. The structure of AHA, which is the first experimentally determined three-dimensional structure of a psychrophilic enzyme, resembles those of other known alpha-amylases of various origins with a surprisingly greatest similarity to mammalian alpha-amylases. AHA contains a chloride ion which activates the hydrolytic cleavage of substrate alpha-1,4-glycosidic bonds. The chloride binding site is situated approximately 5 angstroms from the active site which is characterized by a triad of acid residues (Asp 174, Glu 200, Asp 264). These are all involved in firm binding of the Tris moiety. A reaction mechanism for substrate hydrolysis is proposed on the basis of the Tris inhibitor binding and the chloride activation. A trio of residues (Ser 303, His 337, Glu 19) having a striking spatial resemblance with serine-protease like catalytic triads was found approximately 22 angstroms from the active site. We found that this triad is equally present in other chloride dependent alpha-amylases, and suggest that it could be responsible for autoproteolytic events observed in solution for this cold adapted alpha-amylase. PMID:9541387
Improvement of Starch Digestion Using ?-Amylase Entrapped in Pectin-Polyvinyl Alcohol Blend
Cruz, Maurício; Fernandes, Kátia; Cysneiros, Cristine; Nassar, Reginaldo; Caramori, Samantha
Polyvinyl alcohol (PVA) and pectin blends were used to entrap ?-amylase (Termamyl) using glutaraldehyde as a cross-linker. The effect of glutaraldehyde concentration (0.25, 0.5, 0.75, 1.0, and 1.25%) on the activity of the immobilized enzyme and rate of enzyme released was tested during a 24?h period. Characteristics of the material, such as scanning electron microscopy (SEM), tensile strength (TS), elongation, and rate of dissolution in water (pH 5.7), ruminal buffering solution (pH 7.0), and reactor containing 0.1?mol?L?1 sodium phosphate buffer (pH 6.5), were also analyzed. SEM results showed that the surfaces of the pectin/PVA/amylase films were highly irregular and rough. TS values increased as a function of glutaraldehyde concentration, whereas percentage of elongation (%E) decreased. Pectin/PVA/amylase films presented similar values of solubility in the tested solvents. The material obtained with 0.25% glutaraldehyde performed best with repeated use (active for 24?h), in a phosphate buffer reactor. By contrast, the material obtained with 1.25% glutaraldehyde presented higher performance during in vitro testing using an artificial rumen. The results suggest that pectin/PVA/amylase is a highly promising material for biotechnological applications. PMID:25949991
Interaction mechanism between green tea extract and human ?-amylase for reducing starch digestion.
Miao, Ming; Jiang, Bo; Jiang, Huan; Zhang, Tao; Li, Xingfeng
This study evaluated the inhibitory effects of the green tea extract on human pancreatic ?-amylase activity and its molecular mechanism. The green tea extract was composed of epicatechin (59.2%), epigallocatechin gallate (14.6%) and epicatechin gallate (26.2%) as determined by HPLC analysis. Enzyme activity measurement showed that % inhibition and IC50 of the green tea extract (10%, based on starch) were 63.5% and 2.07mg/ml, respectively. The Michaelis-Menten constant remained unchanged but the maximal velocity decreased from 0.43 (control) to 0.07mg/(ml×min) (4mg/ml of the green tea extract), indicating that the green tea extract was an effective inhibitor against ?-amylase with a non-competitive mode. The fluorescence data revealed that the green tea extract bound with ?-amylase to form a new complex with static quenching mechanism. Docking study showed the epicatechin gallate in the green tea extract presented stronger affinity than epigallocatechin gallate, with more number of amino acid residues involved in amylase binding with hydrogen bonds and Van der Waals forces. Thus, the green tea extract could be used to manipulate starch digestion for potential health benefits. PMID:25976786
Attenuation of Porphyromonas gingivalis oral infection by ??amylase and pentamidine.
Li, Ying; Miao, Yu-Song; Fu, Yun; Li, Xi-Ting; Yu, Shao-Jie
The Porphyromonas gingivalis bacterium is one of the most influential pathogens in oral infections. In the current study, the antimicrobial activity of ??amylase and pentamidine against Porphyromonas gingivalis was evaluated. Their in vitro inhibitory activity was investigated with the agar overlay technique, and the minimal inhibitory and bactericidal concentrations were determined. Using the bactericidal concentration, the antimicrobial actions of the inhibitors were investigated. In the present study, multiple techniques were utilized, including scanning electron microscopy (SEM), general structural analysis and differential gene expression analysis. The results obtained from SEM and bactericidal analysis indicated a notable observation; the pentamidine and ??amylase treatment destroyed the structure of the bacterial cell membranes, which led to cell death. These results were used to further explore these inhibitors and the mechanisms by which they act. Downregulated expression levels were observed for a number of genes coding for hemagglutinins and gingipains, and various genes involved in hemin uptake, chromosome replication and energy production. However, the expression levels of genes associated with iron storage and oxidative stress were upregulated by ??amylase and pentamidine. A greater effect was noted in response to pentamidine treatment. The results of the present study demonstrate promising therapeutic potential for ??amylases and pentamidine. These molecules have the potential to be used to develop novel drugs and broaden the availability of pharmacological tools for the attenuation of oral infections caused by Porphyromonas gingivalis. PMID:25846026
alpha-Amylase production by Bacillus subtilis with dregs in an external-loop airlift bioreactor.
Yuguo; Zhao; Xiaolong
An external-loop airlift bioreactor, with a low ratio 2.9 of height-to-diameter of the riser and a ratio 6.6 of riser-to-downcomer diameter, was used to produce alpha-amylase from fermentation with dregs by Bacillus subtilis. The effects of gas flow rate and liquid volume on alpha-amylase production were investigated. After a 36-h fermentation time, an average of 432.3U/ml alpha-amylase activity was obtained under the conditions of liquid volume 8.5l and gas flow rate 1.2vvm for the first 12h of fermentation, 1.4vvm from 12 to 27h, and 1.2vvm from 27h to the end. The activity was higher than that obtained in shaking flasks (409.0U/ml) and in a mechanically stirred tank bioreactor (397.2U/ml) under optimized operating conditions. The fermentation cycle of the airlift bioreactor was shorter than the 48h required for the shaking flasks and close to the 36h of the mechanically stirred tank bioreactor. It was demonstrated that the external-loop airlift bioreactor could substitute for the traditional mechanically stirred tank bioreactor to produce alpha-amylase from fermentation by Bacillus subtilis with dregs. PMID:10817816
Influence of substratum hydrophobicity on salivary pellicles: organization or composition?
Aroonsang, Watcharapong; Sotres, Javier; El-Schich, Zahra; Arnebrant, Thomas; Lindh, Liselott
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
Lapatinib in Treating Patients With Recurrent and/or Metastatic Adenoid Cystic Cancer or Other Salivary Gland Cancers
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
Microencapsulation of purified amylase enzyme from pitaya (Hylocereus polyrhizus) peel in Arabic gum-chitosan using freeze drying.
Amid, Mehrnoush; Manap, Yazid; Zohdi, Nor Khanani
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
21 CFR 184.1012 - ?-Amylase enzyme preparation from Bacillus stearothermophilus.
Code of Federal Regulations, 2014 CFR
...2014-04-01 false α-Amylase enzyme preparation from Bacillus stearothermophilus...as GRAS § 184.1012 ?-Amylase enzyme preparation from Bacillus stearothermophilus. (a) ?-Amylase enzyme preparation is obtained from...
21 CFR 184.1012 - ?-Amylase enzyme preparation from Bacillus stearothermophilus.
Code of Federal Regulations, 2011 CFR
...2011-04-01 false α-Amylase enzyme preparation from Bacillus stearothermophilus...as GRAS § 184.1012 ?-Amylase enzyme preparation from Bacillus stearothermophilus. (a) ?-Amylase enzyme preparation is obtained from...
21 CFR 184.1012 - ?-Amylase enzyme preparation from Bacillus stearothermophilus.
Code of Federal Regulations, 2010 CFR
...2009-04-01 true α-Amylase enzyme preparation from Bacillus stearothermophilus...as GRAS § 184.1012 ?-Amylase enzyme preparation from Bacillus stearothermophilus. (a) ?-Amylase enzyme preparation is obtained from...
Management of salivary gland malignancies: current and developing therapies
Mark AgulnikCamille; Camille F. McGann; Bharat B. Mittal; Sara C. Gordon; Joel B. Epstein
Salivary gland tumors are rare, clinically diverse neoplasms that represent less than 1% of all malignancies. In locoregional\\u000a recurrent or metastatic disease, systemic therapy is the standard approach. While numerous small phase II studies have evaluated\\u000a the activity of cytotoxic agents, either alone or in combination, the response rates are generally modest with objective response\\u000a rates ranging from 15%–50%. Duration
Barley ?-amylase\\/subtilisin inhibitor. I. Isolation and characterization
John Mundy; IB Svendsen; Jørn Hejgaard
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
Abdominal pain and a raised amylase? It's not always pancreatitis. . .
Oluwatowoju, I O; Abu, O E; Lawson, G
We report the case of a 72 year old man with a history of COPD and heavy alcohol consumption who was initially diagnosed with acute pancreatitis based on a presentation with epigastric pain and elevated serum amylase. Review of his notes revealed several previous similar admissions and extensive normal investigations apart from persistently elevated amylase. Further analysis showed evidence of macroamylasaemia which accounted for the apparently high serum amylase level. PMID:24098876
Plasma amylase estimation in recurrent abdominal pain in children.
Wheeler, R. A.; Colquhoun-Flannery, W. A.; Johnson, C. D.
In a prospective study of 50 children who were admitted on more than one occasion with undiagnosed abdominal pain, the serum amylase was found to be normal in every case. One case of acute pancreatitis was diagnosed in over 8000 admissions. Serum amylase estimation did not contribute to the management of children with recurrent abdominal pain, and acute pancreatitis is so rare that routine amylase estimations cannot be recommended in paediatric surgical practice. PMID:1384415
Production of ?-amylase for the biosynthesis of gold nanoparticles using Streptomyces sp. MBRC-82.
Manivasagan, Panchanathan; Venkatesan, Jayachandran; Kang, Kyong-Hwa; Sivakumar, Kannan; Park, Sun-Joo; Kim, Se-Kwon
Marine actinobacterial synthesis of gold nanoparticles has good potential to develop simple, cost-effective and eco-friendly methods for production of important biomaterials. In this context, gold nanoparticles have attracted considerable attention in recent years, owing to their various applications. In this paper, we report on the production of ?-amylase for the extracellular synthesis of gold nanoparticles using Streptomyces sp. MBRC-82. Medium composition and culture conditions for ?-amylase production were statistically optimized. Plackett-Burman design was employed to find out the optimal medium constituents and culture conditions to enhance ?-amylase production. Box-Behnken design revealed that three independent variables namely soluble starch (5.8484 g), peptone (3.5191 g), and NaCl (0.3829) significantly influenced ?-amylase production. The gold nanoparticles were characterized by ultraviolet-visible (UV-vis) spectrometer, X-ray diffractometer (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDXA), and transmission electron microscopy (TEM). The particles synthesized using the optimized enzyme activity ranged from 20 to 80 nm with an average particle size of 40 nm and therefore can be extended to various medicinal applications. PMID:25128097
Biochemical characterization of an extracellular polyextremophilic ?-amylase from the halophilic archaeon Halorubrum xinjiangense.
Moshfegh, Mahsa; Shahverdi, Ahmad Reza; Zarrini, Gholamreza; Faramarzi, Mohammad Ali
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
Relation of diagnostic serum amylase levels to aetiology and severity of acute pancreatitis.
Winslet, M; Hall, C; London, N J; Neoptolemos, J P
The sensitivity of diagnostic serum amylase (greater than 1000 iu/l) was assessed in 417 patients with acute pancreatitis as a result of gall stones (258), alcohol (104), or miscellaneous causes (55), of whom 111 (27%) had a clinically severe attack (including 34 deaths). On hospital admission, an amylase value diagnostic of pancreatitis was found in 96.1% of all mild cases and in 87.4% of severe cases (p less than 0.001); at 48 hours these values were 33.3% and 48.2% respectively (p = 0.026). Diagnostic amylase levels for alcoholic patients were found in 86% of mild cases on admission and in 76% of severe cases (p less than 0.001, compared with other groups). The diagnostic levels were also significantly lower at 24 hours for both the alcoholic and miscellaneous groups compared with the gall stone group (p less than 0.001). Eight of 27 (30%) patients with a serum amylase activity less than 1000 iu/l had pancreatic necrosis compared with 12 of the remaining 390 (3.1%) patients (p less than 0.001); the mortality was also significantly different (44% v 5.6% respectively, p less than 0.001). These data support the view that more sensitive tests for acute pancreatitis are needed for routine use especially in those whose disease has an alcoholic aetiology. PMID:1379569
Codon Optimization Significantly Improves the Expression Level of ?-Amylase Gene from Bacillus licheniformis in Pichia pastoris
Wang, Jian-Rong; Li, Yang-Yuan; Liu, Dan-Ni; Liu, Jing-Shan; Li, Peng; Chen, Li-Zhi; Xu, Shu-De
?-Amylase as an important industrial enzyme has been widely used in starch processing, detergent, and paper industries. To improve expression efficiency of recombinant ?-amylase from Bacillus licheniformis (B. licheniformis), the ?-amylase gene from B. licheniformis was optimized according to the codon usage of Pichia pastoris (P. pastoris) and expressed in P. pastoris. Totally, the codons encoding 305 amino acids were optimized in which a total of 328 nucleotides were changed and the G+C content was increased from 47.6 to 49.2%. The recombinants were cultured in 96-deep-well microplates and screened by a new plate assay method. Compared with the wild-type gene, the optimized gene is expressed at a significantly higher level in P. pastoris after methanol induction for 168?h in 5- and 50-L bioreactor with the maximum activity of 8100 and 11000?U/mL, which was 2.31- and 2.62-fold higher than that by wild-type gene. The improved expression level makes the enzyme a good candidate for ?-amylase production in industrial use. PMID:26171389
Iron binding modulates candidacidal properties of salivary histatin 5.
Puri, S; Li, R; Ruszaj, D; Tati, S; Edgerton, M
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
Partial purification and some properties of alpha-amylase from Bacillus subtilis KIBGE-HAS.
Bano, Saeeda; Ul Qader, Shah Ali; Aman, Afsheen; Azhar, Abid
An extracellular alpha-amylase from Bacillus subtilis KIBGE-HAS was partially purified by ultrafiltration and ammonium sulphate precipitation with 19.2-fold purification and specific activity of 4195 U/mg. The enzyme showed relatively high thermostability and retained 62% of its activity when kept at 70 degrees C for 15 min. alpha-Amylase was highly stable at -18 degrees C and loss of activity was very low during stability study. Metal ions like Mn2+ Ca2+, Co2+, K+, Mg2+, and Fe3+ activated the enzyme, while Hg2+ Ba2+, Cu2+, Na+ and Al3+ strongly inhibited the activity. The a-amylase was highly stable in various surfactants and detergents. In the presence of surfactants such as SDS and Triton X-100 the enzyme activity was found 2.9 and 1.8-fold higher respectively than control. The non-ionic detergents (Tween 20 and Tween 80) exhibited slightly inhibitory effect on the enzyme activity. PMID:20027871
Inhibitory effects of medicinal mushrooms on ?-amylase and ?-glucosidase - enzymes related to hyperglycemia.
Su, Chun-Han; Lai, Min-Nan; Ng, Lean-Teik
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
Enhanced Production and Characterization of a Solvent Stable Amylase from Solvent Tolerant Bacillus tequilensis RG-01: Thermostable and Surfactant Resistant
Tiwari, Soni; Shukla, Neha; Mishra, Pooja; Gaur, Rajeeva
Ten bacterial strains isolated from the soil samples in the presence of cyclohexane were screened for amylase production. Among them, culture RG-01 was adjudged as the best amylase producer and was identified as Bacillus tequilensis from MTCC, Chandigarh. The isolate showed maximum amylase production (8100?U/mL) in the presence of starch, peptone, and Ca2+ ions at 55°C pH 7.0 within 24?h of incubation. The enzyme was stable in the presence of n-dodecane, isooctane, n-decane, xylene, toluene, n-hexane, n-butanol, and cyclohexane, respectively. The presence of benzene, methanol, and ethanol marginally reduced the amylase stability, respectively. The enzyme was showed it 100% activity at 55°C and pH 7.0 with 119% and 127% stability at 55°C and pH 7.0, respectively. The enzyme was also stable in the presence of SDS, Tween-40, Tween-60, and Tween-80 (1%) and was found stimulatory effect, respectively. Only Triton-X-100 showed a moderate inhibitory effect (5%) on amylase activity. This isolate (Bacillus tequilensis RG-01) may be useful in several industrial applications owing to its thermotolerant and organic solvents and surfactants resistance characteristics. PMID:25401163
Digestive alpha-amylases of the flour moth Ephestia kuehniella--adaptation to alkaline environment and plant inhibitors.
Pytelková, Jana; Hubert, Jan; Lepsík, Martin; Sobotník, Jan; Sindelka, Radek; Krízková, Iva; Horn, Martin; Mares, Michael
The digestive tract of lepidopteran insects is extremely alkaline. In the present work, molecular adaptation of amylolytic enzymes to this environment was investigated in the flour moth Ephestia kuehniella, an important stored-product pest. Three digestive alpha-amylases [Ephestia kuehniella alpha-amylase isoenzymes 1-3 (EkAmy1-3)] with an alkaline pH optimum were purified from larvae and biochemically characterized. These isoenzymes differ significantly in their sensitivity to alpha-amylase inhibitors of plant origin that are directed against herbivores as antifeedants. Such functional variability renders the amylolytic system less vulnerable to suppression by plant defensive molecules. Moreover, we found that expression of alpha-amylases is upregulated in larvae feeding on a diet enriched with an alpha-amylase inhibitor. The alpha-amylases are secreted into the larval midgut by an exocytotic mechanism, as revealed by immunogold microscopy. The cDNA sequence of EkAmy3 was determined, and a homology model of EkAmy3 was built in order to analyze the structural features responsible for adaptation to alkaline pH. First, the overall fold was found to be stabilized by remodeling of ion pairs. Second, molecular simulations supported by activity measurements showed that EkAmy3 does not bind a Cl(-), owing to an Arg-to-Gln mutation in a conserved binding site. The Cl(-)-binding residues are in contact with the catalytic residues, and this change might help to fine-tune the catalytic pK(a) values to an alkaline pH optimum. We conclude that lepidopteran alpha-amylases are evolutionarily adapted in terms of structure and expression dynamics for effective functioning in the digestive system. PMID:19476481
Cloning and expression of a Bacillus coagulans amylase gene in Escherichia coli
Pierre Cornelis; Colette Digneffe; Karine Willemot
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;
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
Tsuji, Akihiko; Nishiyama, Nami; Ohshima, Miki; Maniwa, Saori; Kuwamura, Shuji; Shiraishi, Masataka; Yuasa, Keizo
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
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.
Tsuji, Akihiko; Nishiyama, Nami; Ohshima, Miki; Maniwa, Saori; Kuwamura, Shuji; Shiraishi, Masataka; Yuasa, Keizo
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
Glycoinositolphospholipids from Trypanosomatids Subvert Nitric Oxide Production in Rhodnius prolixus Salivary Glands
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
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
Ratios of plasma and salivary testosterone throughout puberty: production versus bioavailability.
Rilling, J K; Worthman, C M; Campbell, B C; Stallings, J F; Mbizva, M
Because diffusion of testosterone (T) into the salivary gland is thought to be largely limited to the free, biologically active fraction, salivary testosterone is expected to provide a better measure of testosterone bioavailability in the body than is plasma testosterone. Matched saliva and blood spot samples were collected from 218 Zimbabwean males (age 11-23) who were at different stages of puberty, as assessed by self-reported Tanner genital stage ratings. Testosterone concentrations in these matched samples were highly correlated (r = 0.83). Both salivary and plasma testosterone (converted from blood spot value) showed expected significant increases across puberty. However, plasma testosterone distinguished among subjects at different stages of genital development more effectively than did salivary testosterone, suggesting the former to be a better marker of testosterone bioavailability. Sex hormone-binding globulin (SHBG) levels were also measured in a subgroup of 93 of these subjects. After controlling for plasma T concentrations, we found a small but significant inverse correlation between blood spot SHBG levels and the proportion of plasma testosterone recovered in salvia, supporting the hypothesis that SHBG-related changes in T bioavailability are detectable in saliva. We conclude that salivary testosterone accurately reflects testicular production of testosterone, but that neither salivary testosterone nor plasma testosterone is clearly superior to the other as a measure of testosterone bioavailability. PMID:8776800
Pupil and Salivary Indicators of Autonomic Dysfunction in Autism Spectrum Disorder
Anderson, Christa J.; Colombo, John; Unruh, Kathryn E.
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
Salivary alcohol dehydrogenase in non-smoking and smoking alcohol-dependent persons.
Waszkiewicz, Napoleon; Jelski, Wojciech; Zalewska, Anna; Szulc, Agata; Szmitkowski, Maciej; Zwierz, Krzysztof; Szajda, S?awomir Dariusz
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
Salivary stones: symptoms, aetiology, biochemical composition and treatment.
Kraaij, S; Karagozoglu, K H; Forouzanfar, T; Veerman, E C I; Brand, H S
Salivary stones, also known as sialoliths, are calcified concrements in the salivary glands. Sialoliths are more frequently located in the submandibular gland (84%), than in the parotid gland (13%). The majority of the submandibular stones are located in Wharton's duct (90%), whereas parotid stones are more often located in the gland itself. Salivary stones consist of an amorphous mineralised nucleus, surrounded by concentric laminated layers of organic and inorganic substances. The organic components of salivary stones include collagen, glycoproteins, amino acids and carbohydrates. The major inorganic components are hydroxyapatite, carbonate apatite, whitlockite and brushite. The management of salivary stones is focused on removing the salivary stones and preservation of salivary gland function which depends on the size and location of the stone. Conservative management of salivary stones consists of salivary gland massage and the use of sialogogues. Other therapeutic options include removal of the stone or in some cases surgical removal of the whole salivary gland. PMID:25476659
Enzyme Assay Guided Isolation of an ?-Amylase Inhibitor Flavonoid from Vaccinium arctostaphylos Leaves
Nickavar, Bahman; Amin, Gholamreza
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
Enzyme Assay Guided Isolation of an ?-Amylase Inhibitor Flavonoid from Vaccinium arctostaphylos Leaves.
Nickavar, Bahman; Amin, Gholamreza
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
Optimised amylases extraction from oat seeds and its impact on bread properties.
Ben Halima, Nihed; Borchani, Maha; Fendri, Imen; Khemakhem, Bassem; Gosset, David; Baril, Patrick; Pichon, Chantal; Ayadi, Mohamed-Ali; Abdelkafi, Slim
Statistical approaches were employed for the optimisation of the extraction of amylolytic activity from oat (Avena sativa) seeds. The application of the response surface methodology allows us to determine a set of optimal conditions (ratio seed weight/buffer volume 0.1, germination days 10 days, temperature 20 °C and pH 5.6). Experiments carried out under these conditions led to amylase production yield of 91 U/g. Its maximal activity was in the pH 5.6 and at 55 °C. Study of the incorporation of the optimised oat extract into the bread formulation revealed an improvement of the sensory quality and the textural properties of fresh and stored bread. Three-dimensional elaborations of Confocal Laser Scanning Microscopy (CLSM) images were performed on crumb of the different breads to evaluate the influence of amylase activity on microstructure. The result showed improved baking characteristics as well as overall microscopic and macroscopic appearance. PMID:25453287
Disruption of amylase genes by RNA interference affects reproduction in the Pacific oyster Crassostrea gigas.
Huvet, Arnaud; Béguel, Jean-Philippe; Cavaleiro, Nathalia Pereira; Thomas, Yoann; Quillien, Virgile; Boudry, Pierre; Alunno-Bruscia, Marianne; Fabioux, Caroline
Feeding strategies and digestive capacities can have important implications for variation in energetic pathways associated with ecological and economically important traits, such as growth or reproduction in bivalve species. Here, we investigated the role of amylase in the digestive processes of Crassostrea gigas, using in vivo RNA interference. This approach also allowed us to investigate the relationship between energy intake by feeding and gametogenesis in oysters. Double-stranded (ds)RNA designed to target the two ?-amylase genes A and B was injected in vivo into the visceral mass of oysters at two doses. These treatments caused significant reductions in mean mRNA levels of the amylase genes: -50.7% and -59% mRNA A, and -71.9% and -70.6% mRNA B in 15 and 75?µg dsRNA-injected oysters, respectively, relative to controls. Interestingly, reproductive knock-down phenotypes were observed for both sexes at 48?days post-injection, with a significant reduction of the gonad area (-22.5% relative to controls) and germ cell under-proliferation revealed by histology. In response to the higher dose of dsRNA, we also observed reductions in amylase activity (-53%) and absorption efficiency (-5%). Based on these data, dynamic energy budget modeling showed that the limitation of energy intake by feeding that was induced by injection of amylase dsRNA was insufficient to affect gonadic development at the level observed in the present study. This finding suggests that other driving mechanisms, such as endogenous hormonal modulation, might significantly change energy allocation to reproduction, and increase the maintenance rate in oysters in response to dsRNA injection. PMID:25883379
A nested alpha-amylase gene in Drosophila ananassae.
Da Lage, Jean-Luc; Maisonhaute, Claude; Maczkowiak, Frédérique; Cariou, Marie-Louise
The amylase gene family of Drosophila ananassae consists in seven copies, scattered on several chromosomal arms. We have evidenced that a member of the family, Amy35, lies within an intron of a gene homologous to the CG14696 gene of D. melanogaster. This nested arrangement seems restricted to the D. ananassae subgroup. The nested and the nest genes are encoded on opposite strands. Both are actively transcribed in the midgut at the same time, raising the possibility of interference between their mRNAs. Our data also help to elucidate the history of the Amy family, suggesting that Amy35 arose by duplication and translocation from another ancestral locus, into a formerly short intron, in an ancestor of the subgroup. PMID:14629045
Transglycosylation of neohesperidin dihydrochalcone by Bacillus stearothermophilus maltogenic amylase.
Cho, J S; Yoo, S S; Cheong, T K; Kim, M J; Kim, Y; Park, K H
Neohesperidin dihydrochalcone (NHDC), a sweet compound derived from citrus fruits, was modified to a series of its oligosaccharides by transglycosylation activity of Bacillus stearothermophilus maltogenic amylase (BSMA). Maltotriose as a donor was reacted with NHDC as an acceptor to glycosylate for the purpose of increasing the solubility of NHDC. Maltosyl-NHDC was a major transglycosylation product among the several transfer products by TLC analysis. The structure of the major transglycosylation product was determined to be maltosyl-alpha-(1,6)-neohesperidin dihydrochalcone by MALDI-TOF/MS and (1)H and (13)C NMR. Maltosyl-NHDC was 700 times more soluble in water and 7 times less sweet than NHDC. PMID:10691608
Antioxidant and ?-Amylase Inhibitory Property of Phyllanthus virgatus L.: An In Vitro and Molecular Interaction Study
Hashim, Arshya; Khan, M. Salman; Khan, Mohd. Sajid; Baig, Mohd. Hassan
The present study on Phyllanthus virgatus, known traditionally for its remedial potential, for the first time provides descriptions of the antioxidant and inhibition of ?-amylase enzyme activity first by in vitro analyses, followed by a confirmatory in silico study to create a stronger biochemical rationale. Our results illustrated that P. virgatus methanol extract exhibited strong antioxidant and oxidative DNA damage protective activity than other extracts, which was well correlated with its total phenolic content. In addition, P. virgatus methanol extract strongly inhibited the ?-amylase activity (IC50 33.20 ± 0.556??g/mL), in a noncompetitive manner, than acarbose (IC50 76.88 ± 0.277??g/mL), which showed competitive inhibition. Moreover, this extract stimulated the glucose uptake activity in 3T3-L1 cells and also showed a good correlation between antioxidant and ?-amylase activities. The molecular docking studies of the major bioactive compounds (9,12-octadecadienoic acid, asarone, 11-octadecenoic acid, and acrylic acid) revealed via GC-MS analysis from this extract mechanistically suggested that the inhibitory property may be due to the synergistic effect of these bioactive compounds. These results provide substantial basis for the future use of P. virgatus methanol extract and its bioactive compound in in vivo system for the treatment and management of diabetes as well as in the related condition of oxidative stress. PMID:23957001
Analysis of salivary gland transcripts of the sand fly Lutzomyia ayacuchensis, a vector of Andean-type cutaneous leishmaniasis
Kato, Hirotomo; Jochim, Ryan C.; Gomez, Eduardo A.; Uezato, Hiroshi; Mimori, Tatsuyuki; Korenaga, Masataka; Sakurai, Tatsuya; Katakura, Ken; Valenzuela, Jesus G.; Hashiguchi, Yoshihisa
The saliva of blood sucking insects contains potent pharmacologically active components that assist them in counteracting the host hemostatic and inflammatory systems during blood feeding. In addition, sand fly salivary proteins affect host immunity and have the potential to be a vaccine against Leishmania infection. In the present study, the salivary gland transcripts of Lutzomyia ayacuchensis, a vector of cutaneous leishmaniasis in Ecuadorian and Peruvian Andes, were analyzed by sequencing randomly selected clones of the salivary gland cDNA library of this sand fly. This resulted in the identification of the most abundant transcripts coding for secreted proteins. These proteins were homologous to the salivary molecules present in other sand flies including the RGD-containing peptide, PpSP15/SL1 family protein, yellow-related protein, putative apyrase, antigen 5-related protein, D7 family protein, and 27 kDa salivary protein. Of note, homologues of maxadilan, an active vasodilator abundantly present in saliva of Lutzomyia longipalpis, were not identified. This analysis is the first description of salivary proteins from a sand fly of the subgenus Helcocyrtomyia and from vector of cutaneous leishmaniasis in the New World. The present analysis will provide further insights into the evolution of salivary components in blood sucking arthropods. PMID:23000112
Oral integrity and salivary profile in myeloma patients undergoing high-dose therapy followed by autologous SCT.
Avivi, I; Avraham, S; Koren-Michowitz, M; Zuckerman, T; Aviv, A; Ofran, Y; Benyamini, N; Nagler, A; Rowe, J M; Nagler, R M
The underlying mechanism of high-dose therapy (HDT)-related oral mucositis (OM) may be partly mediated by alterations in the normal salivary composition. This study evaluated salivary antioxidant and immunological capacities observed in myeloma patients suffering from HDT-related OM, and assessed potential contribution of these factors to OM development. Twenty-five consecutive myeloma patients treated with melphalan 200 mg/m(2) followed by autologous SCT were enrolled. Patients underwent a daily assessment for OM, and salivary samples were collected on days -3 and +7 of transplantation and analyzed for secretory IgA and antioxidant capacity. The degree of mucosal damage was assessed by measuring the salivary carbonyl and albumin (Alb) levels. OM, reported in 96% of patients, appeared to be most severe on 8 day after transplantation (range: +2 to +14). Clinical mucositis was associated with significant reduction in salivary secretory IgA (54%; P=0.05), and antioxidant activity, measured by total antioxidant status (40%; P=0.0004), antioxidant capacity (ImAnOx) (23%; P=0.002) and uric acid level (51%; P=0.006). The increase found in salivary Alb (119%; P=0.024) and carbonyl (28%; P=0.047) levels, indicates mucosal and oxidative damage, respectively. These salivary changes might enhance mucositis development and symptoms. Therapeutic interventions, enhancing antioxidative and immunological activities need to be investigated. PMID:19029961
Induction of a Mitosis Delay and Cell Lysis by High-Level Secretion of Mouse ?-Amylase from Saccharomyces cerevisiae
Wang, Bi-Dar; Kuo, Tsong-Teh
Some foreign proteins are produced in yeast in a cell cycle-dependent manner, but the cause of the cell cycle dependency is unknown. In this study, we found that Saccharomyces cerevisiae cells secreting high levels of mouse ?-amylase have elongated buds and are delayed in cell cycle completion in mitosis. The delayed cell mitosis suggests that critical events during exit from mitosis might be disturbed. We found that the activities of PP2A (protein phosphatase 2A) and MPF (maturation-promoting factor) were reduced in ?-amylase-oversecreting cells and that these cells showed a reduced level of assembly checkpoint protein Cdc55, compared to the accumulation in wild-type cells. MPF inactivation is due to inhibitory phosphorylation on Cdc28, as a cdc28 mutant which lacks an inhibitory phosphorylation site on Cdc28 prevents MPF inactivation and prevents the defective bud morphology induced by overproduction of ?-amylase. Our data also suggest that high levels of ?-amylase may downregulate PPH22, leading to cell lysis. In conclusion, overproduction of heterologous ?-amylase in S. cerevisiae results in a negative regulation of PP2A, which causes mitotic delay and leads to cell lysis. PMID:11472949
Salivary immunoglobulins and prevalent coronary artery disease.
Janket, S; Meurman, J H; Baird, A E; Qvarnström, M; Nuutinen, P; Ackerson, L K; Hong, J; Muthukrishnan, P; Van Dyke, T E
Previous studies examined the serum immunoglobulin levels in relation to coronary artery disease (CAD). We hypothesized that the salivary immunoglobulins might better estimate oral infections in this relationship. Multivariate logistic regression analyses utilizing the data from 256 angiographically confirmed CAD patients and 250 non-CAD individuals that controlled for age, sex, smoking, diabetes, total/HDL cholesterol ratio, hypertension, and education revealed the trends that salivary IgA was positively and salivary IgG was inversely associated with CAD. The odds ratios (OR) of each increasing quartile of salivary IgA were 1.00 (first and second quartiles combined), 1.97, and 1.37 (p-value for trend = 0.06), while those for salivary IgG were 1.00, 0.77, 0.60, and 0.51 (p-value for trend = 0.02). Additionally, salivary IgA correlated positively with C-reactive protein and Asymptotic Dental Score (dental infection score), while IgG was inversely associated with these inflammation markers. Salivary IgA warrants further studies to confirm its role in the risk assessment of CAD. PMID:20177131
Alpha-amylase from germinating soybean (Glycine max) seeds--purification, characterization and sequential similarity of conserved and catalytic amino acid residues.
Kumari, Arpana; Singh, Vinay Kumar; Fitter, Jörg; Polen, Tino; Kayastha, Arvind M
Starch hydrolyzing amylase from germinated soybeans seeds (Glycine max) has been purified 400-fold to electrophoretic homogeneity with a final specific activity of 384 units/mg. SDS-PAGE of the final preparation revealed a single protein band of 100 kDa, whereas molecular mass was determined to be 84 kDa by MALDI-TOF and gel filtration on Superdex-200 (FPLC). The enzyme exhibited maximum activity at pH 5.5 and a pI value of 4.85. The energy of activation was determined to be 6.09 kcal/mol in the temperature range 25-85 degrees C. Apparent Michaelis constant (K(m)((app))) for starch was 0.71 mg/mL and turnover number (k(cat)) was 280 s(-1) in 50 mM sodium acetate buffer, pH 5.5. Thermal inactivation studies at 85 degrees C showed first-order kinetics with rate constant (k) equal to 0.0063 min(-1). Soybean alpha-amylase showed high specificity for its primary substrate starch. High similarity of soybean alpha-amylase with known amylases suggests that this alpha-amylase belongs to glycosyl hydrolase family 13. Cereal alpha-amylases have gained importance due to their compatibility for biotechnological applications. Wide availability and easy purification protocol make soybean as an attractive alternative for plant alpha-amylase. Soybean can be used as commercially viable source of alpha-amylase for various industrial applications. PMID:20655076
Optimization of Amylase Application in Raw Sugar Manufacture. Part I: Characterization of Commercial Alpha-Amylases
Technology Transfer Automated Retrieval System (TEKTRAN)
In recent years there have been warnings by some U.S. refineries that there may be a penalty for high starch concentrations in raw sugar if starch control is not improved. Most commercial amylases used by the U.S. sugar industry to control starch have intermediate temperature stability (up to 85 de...
The Amylase Project: Creating a Classroom of Biotechnologists.
ERIC Educational Resources Information Center
Sweeney, Diane
A biotechnologist-turned-teacher introduces a series of laboratory modules incorporating concepts from microbiology, cellular biology, molecular biology, biochemistry, and evolution. The Amylase Project aims to distill the biotechnology process into a few short steps using amylase, the easiest enzyme to detect of those commonly produced by…
Purification and characterisation of ?-amylase produced by mutant strain of Aspergillus oryzae EMS-18.
Abdullah, Roheena; Ikram-ul-Haq
?-Amylase produced by a mutant strain of Aspergillus oryzae EMS-18 has been purified to homogeneity as judged by sodium dodecyle sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was purified by using 70% ammonium sulphate precipitation followed by anion exchange chromatography on DEAE-Sephadex column and gel filtration on Sephadex G-100. An enzyme purification factor of 9.5-fold was achieved with a final specific activity of 1987.7 U/mg protein and overall yield of 23.8%. The molecular weight of purified ?-amylase was estimated to be 48 kDa by SDS-PAGE. The purified enzyme revealed an optimum assay temperature and pH 40°C and 5.0, respectively. Except Ca(++) all other metal ions such as Mg, Mn, Na, Zn, Ni, Fe, Cu, Co and Ba were found to be inhibitory to enzyme activity. PMID:25424747
Molecular Identification of a Newly Isolated Bacillus subtilis BI19 and Optimization of Production Conditions for Enhanced Production of Extracellular Amylase
Dash, Biplab Kumar; Rahman, M. Mizanur; Sarker, Palash Kumar
A study was carried out with a newly isolated bacterial strain yielding extracellular amylase. The phylogenetic tree constructed on the basis of 16S rDNA gene sequences revealed this strain as clustered with the closest members of Bacillus sp. and identified as Bacillus subtilis BI19. The effect of various fermentation conditions on amylase production through shake-flask culture was investigated. Rice flour (1.25%) as a cheap natural carbon source was found to induce amylase production mostly. A combination of peptone and tryptone as organic and ammonium sulfate as inorganic nitrogen sources gave highest yield. Maximum production was obtained after 24?h of incubation at 37°C with an initial medium pH 8.0. Addition of surfactants like Tween 80 (0.25?g/L) and sodium lauryl sulfate (0.2?g/L) resulted in 28% and 15% increase in enzyme production, respectively. Amylase production was 3.06 times higher when optimized production conditions were used. Optimum reaction temperature and pH for crude amylase activity were 50°C and 6.0, respectively. The crude enzyme showed activity and stability over a fair range of temperature and pH. These results suggest that B. subtilis BI19 could be exploited for production of amylase at relatively low cost and time. PMID:26180814
Effects of Abscisic Acid and Ethylene on the Gibberellic Acid-Induced Synthesis of ?-Amylase by Isolated Wheat Aleurone Layers 1
Varty, Keith; Arreguín, Barbarín L.; Gómez, Miguel T.; López, Pablo Jaime T.; Gómez, Miguel Angel L.
Gibberellic acid-induced ?-amylase synthesis in wheat aleurone layers (Triticum aestivum L. var Potam S-70) escaped from transcriptional control 30 h after addition of the hormone, as evidenced by the tissue's loss of susceptibility to cordycepin. Abscisic acid inhibited the accumulation of ?-amylase activity when added to the tissue during this cordycepin-insensitive phase of enzyme induction. ?-Amylase synthesis was not restored by the addition of cordycepin, indicating that the response to abscisic acid was not dependent upon the continuous synthesis of a short lived RNA. When ethylene was added simultaneously or some time after abscisic acid, the accumulation of ?-amylase activity was sustained or quickly restored. The loss of susceptibility to cordycepin was completely prevented when aleurone layers were incubated with a combination of gibberellic and abscisic acids from the start of the induction period. This effect of abscisic acid was not reversed by ethylene. On the basis of these observations, it is suggested that abscisic acid inhibits both the transcription and translation of ?-amylase mRNA, and that only the latter site of action is susceptible to reversal by ethylene. The rate of incorporation of [methyl-14C]choline into phospholipids was also inhibited by abscisic acid. Ethylene reversed this effect. The effects of abscisic acid and ethylene on phospholipid synthesis were not dependent upon the presence of gibberellic acid. No direct relationship was found between the control of ?-amylase synthesis and membrane formation by abscisic acid and ethylene. PMID:16663284
Sensitive salivary estradiol assay for monitoring ovarian function.
Worthman, C M; Stallings, J F; Hofman, L F
Measurement of steroids in saliva has excited interest because of the numerous potential clinical applications; noninvasive, convenient sampling; and apparently accurate reflection of the concentrations of physiologically active unbound steroid in the circulation. Although assays of saliva for several steroid hormones are available and widely used, assays for salivary estradiol are not, primarily because of methodological limitations. By modifying a commercially available kit for serum estradiol, our laboratory has developed a procedure that is sensitive, highly specific, and reliable for measuring salivary estradiol. Assay sensitivity is 0.5 fmol (0.14 pg; sample concentration 1.3 pmol/L) with a mean interassay CV of 10.8% at low concentrations. Clinical studies showed that values for serum and saliva are highly correlated (P less than 0.001), and demonstrated reliable detection of estradiol peaks during normal ovulatory cycles in serial samples from 15 women. Salivary estradiol peaked at 5.4 (SD 1.9) pmol/L on cycle day 14.4 (SD 3.2), 1.2 (SD 0.8) days before ovulation detected by ultrasound. This assay may be particularly helpful in investigating ovarian function and free estradiol in women at various stages of the reproductive cycle. PMID:2208652
The high maltose-producing ?-amylase of the thermophilic actinomycete, Thermomonospora curvata
Bernadette S. Collins; Catherine T. Kelly; William M. Fogarty; Evelyn M. Doyle
The a-amylase of Thermomonospora curvata catalyses the formation of very high levels of maltose from starch (73%, w\\/w) without the attendant production of glucose. The enzyme was produced extracellularly in high yield during batch fermentation in a 5-1 fermentor. Purification was achieved by ammonium sulphate fractionation, Superose-12 gel filtration and DEAE-Sephacel ionexchange chromatography. The enzyme exhibited maxima for activity at
Functional differences in the acinar cells of the murine major salivary glands.
Kondo, Y; Nakamoto, T; Jaramillo, Y; Choi, S; Catalan, M A; Melvin, J E
In humans, approximately 90% of saliva is secreted by the 3 major salivary glands: the parotid (PG), the submandibular (SMG), and the sublingual glands (SLG). Even though it is known that all 3 major salivary glands secrete saliva by a Cl(-)-dependent mechanism, salivary secretion rates differ greatly among these glands. The goal of this study was to gain insight into the properties of the ion-transporting pathways in acinar cells that might account for the differences among the major salivary glands. Pilocarpine-induced saliva was simultaneously collected in vivo from the 3 major salivary glands of mice. When normalized by gland weight, the amount of saliva secreted by the PG was more than 2-fold larger than that obtained from the SMG and SLG. At the cellular level, carbachol induced an increase in the intracellular [Ca(2+)] that was more than 2-fold larger in PG and SMG than in SLG acinar cells. Carbachol-stimulated Cl(-) efflux and the protein levels of the Ca(2+)-activated Cl(-) channel TMEM16A, the major apical Cl(-) efflux pathway in salivary acinar cells, were significantly greater in PG compared with SMG and SLG. In addition, we evaluated the transporter activity of the Na(+)-K(+)-2Cl(-) cotransporters (NKCC1) and anion exchangers (AE), the 2 primary basolateral Cl(-) uptake mechanisms in acinar cells. The SMG NKCC1 activity was about twice that of the PG and more than 12-fold greater than that of the SLG. AE activity was similar in PG and SLG, and both PG and SLG AE activity was about 2-fold larger than that of SMG. In summary, the salivation kinetics of the 3 major glands are distinct, and these differences can be explained by the unique functional properties of each gland related to Cl(-) movement, including the transporter activities of the Cl(-) uptake and efflux pathways, and intracellular Ca(2+) mobilization. PMID:25680367
Mapping of Barley ( Hordeum vulgare L.) Beta -amylase Alleles in which an Amino Acid Substitution Determines Beta -amylase Isoenzyme Type and the Level of Free Beta -amylase
C.-D. Li; P. Langridge; X.-Q. Zhang; P. E. Eckstein; B. G. Rossnagel; R. C. M. Lance; E. B. Lefol; M.-Y. Lu; B. L. Harvey; G. J. Scoles
The three beta -amylase genes (Bmy1, 2 and 3) in cultivated barley were mapped to chromosomes 4HL, 2HL And 4HL respectively using RFLP analysis. No recombinants between Bmy1 andBmy3 were detected among 264 DH lines. Polymorphism of the Sd1 and Sd2 isoenzymes of beta -amylase co-segregated with the Bmy loci on chromosome 4HL in a doubled-haploid population of the cross
The concept of the ?-amylase family: Structural similarity and common catalytic mechanism
Takashi Kuriki; Tadayuki Imanaka
This review reconsiders the concept of the ?-amylase family in the light of the recent wealth of information on the structures, the catalytic mechanisms, and the classification of amylases. We proposed a general concept for an enzyme family, the ?-amylase family including most of the amylases and related enzymes in 1992, based on the structural similarity and the common catalytic
Oncocytoma of palatal minor salivary gland.
Motallebnejad, Mina; Seyedmajidi, Maryam; Khakbaz Baboli, Oveis; Yarmand, Fateme
Oncocytoma is a rare benign salivary gland tumor, which mostly occurs in the parotid gland. In this article, we describe an early onset of oncocytoma of minor salivary gland in a 36-year, white male. On clinical examination, we encounter with a painless, granular, sessile mass. After Excisional biopsy, the histopathological features revealed sheets of cells with abundant granular eosinophilic cytoplasm, and large, round nuclei that are known as "Oncocyte". PMID:25959915
Fermentative Production and Thermostability Characterization of ? Amylase from Aspergillus Species and Its Application Potential Evaluation in Desizing of Cotton Cloth
Chimata, Murali Krishna; Chetty, Chellu S.; Suresh, Challa
The production of extracellular amylase was investigated employing our laboratory isolate, Aspergillus niger sp. MK 07 and effect of process variables on enzyme production, was studied in a fermentor. It was found that amylase production was maximum when the fermentor volume was maintained at 70%, rate of agitation at 250?rpm, air supply at 2.5?vvm, inoculum concentration of 10%, and a pH of 5.0. Highest enzyme production obtained under all optimized conditions was 1734?U/mL with sucrose as carbon substrate and corn steep liquor as nitrogen source. Enzyme purification studies by ammonium sulphate precipitation and Sephadex G-100 chromatography was evaluated for obtaining purified enzyme. Thermostability of amylase were evaluated with varying concentrations from 0.2 to 0.5?M concentrations of calcium chloride and the highest activity obtained was 3115?U/mL with 0.3?M calcium chloride at 55°C. Effect of temperature and pH on the activity of purified enzyme was evaluated and the purified enzyme showed an activity till 75°C and a pH of 6.5. Application potential of partially purified alpha amylase on desizing of cotton cloth was evaluated with varying enzyme concentrations from 50 to 500?U/mL and the highest desizing activity was found to be at 300?U/mL. PMID:21977326
Fermentative Production and Thermostability Characterization of ? Amylase from Aspergillus Species and Its Application Potential Evaluation in Desizing of Cotton Cloth.
Chimata, Murali Krishna; Chetty, Chellu S; Suresh, Challa
The production of extracellular amylase was investigated employing our laboratory isolate, Aspergillus niger sp. MK 07 and effect of process variables on enzyme production, was studied in a fermentor. It was found that amylase production was maximum when the fermentor volume was maintained at 70%, rate of agitation at 250?rpm, air supply at 2.5?vvm, inoculum concentration of 10%, and a pH of 5.0. Highest enzyme production obtained under all optimized conditions was 1734?U/mL with sucrose as carbon substrate and corn steep liquor as nitrogen source. Enzyme purification studies by ammonium sulphate precipitation and Sephadex G-100 chromatography was evaluated for obtaining purified enzyme. Thermostability of amylase were evaluated with varying concentrations from 0.2 to 0.5?M concentrations of calcium chloride and the highest activity obtained was 3115?U/mL with 0.3?M calcium chloride at 55°C. Effect of temperature and pH on the activity of purified enzyme was evaluated and the purified enzyme showed an activity till 75°C and a pH of 6.5. Application potential of partially purified alpha amylase on desizing of cotton cloth was evaluated with varying enzyme concentrations from 50 to 500?U/mL and the highest desizing activity was found to be at 300?U/mL. PMID:21977326
Interactions between developing nerves and salivary glands.
Ferreira, João N; Hoffman, Matthew P
Our aim is to provide a summary of the field of salivary gland development and regeneration from the perspective of what is known about the function of nerves during these processes. The primary function of adult salivary glands is to produce and secrete saliva. Neuronal control of adult salivary gland function has been a focus of research ever since Pavlov's seminal experiments on salivation in dogs. Less is known about salivary gland innervation during development and how the developing nerves influence gland organogenesis and regeneration. Here, we will review what is known about the communication between the autonomic nervous system and the epithelium of the salivary glands during organogenesis. An important emerging theme is the instructive role of the nervous system on the epithelial stem/progenitor cells during development as well as regeneration after damage. We will provide a brief overview of the neuroanatomy of the salivary glands and discuss recent literature that begins to integrate neurobiology with epithelial organogenesis, which may provide paradigms for exploring these interactions in other organ systems. PMID:23974175
Tobacco plants transformed with the bean. alpha. ai gene express an inhibitor of insect. alpha. -amylase in their seeds. [Nicotiana tabacum; Tenebrio molitor
Altabella, T.; Chrispeels, M.J. (Univ. of California, San Diego, La Jolla (USA))
Bean (Phaseolus vulgaris L.) seeds contain a putative plant defense protein that inhibits insect and mammalian but not plant {alpha}-amylases. We recently presented strong circumstantial evidence that this {alpha}-amylase inhibitor ({alpha}Al) is encoded by an already-identified lectin gene whose product is referred to as lectin-like-protein (LLP). We have now made a chimeric gene consisting of the coding sequence of the lectin gene that encodes LLP and the 5{prime} and 3{prime} flanking sequences of the lectin gene that encodes phytohemagglutinin-L. When this chimeric gene was expressed in transgenic tobacco (Nicotiana tabacum), we observed in the seeds a series of polypeptides (M{sub r} 10,000-18,000) that cross-react with antibodies to the bean {alpha}-amylase inhibitor. Most of these polypeptides bind to a pig pancreas {alpha}-amylase affinity column. An extract of the seeds of the transformed tobacco plants inhibits pig pancreas {alpha}-amylase activity as well as the {alpha}-amylase present in the midgut of Tenebrio molitor. We suggest that introduction of this lectin gene (to be called {alpha}ai) into other leguminous plants may be a strategy to protect the seeds from the seed-eating larvae of Coleoptera.
[Determination of exogenous gamma-amylase residue in honey].
Fei, Xiaoqing; Wu, Bin; Shen, Chongyu; Zhang, Rui; Ding, Tao; Li, Lihua
A novel method for the determination of exogenous gamma-amylase residue in honey using liquid chromatography-isotope ratio mass spectrometry (LC-IRMS) was established. After pre-separation by gel column chromatography, the gamma-amylase in honey samples was separated from the sugars. The gamma-amylase was then used to catalyze maltose into glucose. This enzymatic reaction was under the conditions of 55 degrees C and 0.03 mol/L phosphate buffer solution (pH 4.5) for 48 h. The maltose and glucose in the above enzymatic reaction solution were separated using liquid chromatography. By measuring the content of glucose with isotope ratio mass spectrometry, the gamma-amylase in honey can be determined. The linear range of gamma-amylase was 5 - 200 U/kg with the quantification limit of 5 U/kg. The recoveries were between 89.6% and 108.2% with the relative standard deviations from 3.3% to 4.9%. This method was used to analyze 38 honey and rice syrup samples, and the detection rate of gamma-amylase was 76.3%. To further verify the detection capability of this method, an authentic honey was adulterated with 15% (mass fraction) rice syrup. The gamma-amylase content in this sample was 10.2 U/kg. This method can effectively identify honey adulteration with rice syrups from the perspective of enzymology. PMID:23256379
Monoclinic crystal form of Aspergillus niger ?-amylase in complex with maltose at 1.8?Å resolution
Vuji?i?-Žagar, A.; Dijkstra, B. W.
Aspergillus niger ?-amylase catalyses the hydrolysis of ?-1,4-glucosidic bonds in starch. It shows 100% sequence identity to the A. oryzae homologue (also called TAKA-amylase), three crystal structures of which have been published to date. Two of them belong to the orthorhombic space group P212121 with one molecule per asymmetric unit and one belongs to the monoclinic space group P21 with three molecules per asymmetric unit. Here, the purification, crystallization and structure determination of A. niger ?-amylase crystallized in the monoclinic space group P21 with two molecules per asymmetric unit in complex with maltose at 1.8?Å resolution is reported. Furthermore, a novel 1.6?Å resolution orthorhombic crystal form (space group P21212) of the native enzyme is presented. Four maltose molecules are observed in the maltose–?-amylase complex. Three of these occupy active-site subsites ?2 and ?1, +1 and +2 and the hitherto unobserved subsites +4 (Asp233, Gly234) and +5 (Asp235). The fourth maltose molecule binds at the distant binding sites d1 (Tyr382) and d2 (Trp385), also previously unobserved. Furthermore, it is shown that the active-site groove permits different binding modes of sugar units at subsites +1 and +2. This flexibility of the active-site cleft close to the catalytic centre might be needed for a productive binding of substrate chains and/or release of products. PMID:16880540
Purification and Characterization of a Polyextremophilic ?-Amylase from an Obligate Halophilic Aspergillus penicillioides Isolate and Its Potential for Souse with Detergents
Ali, Imran; Akbar, Ali; Anwar, Mohammad; Prasongsuk, Sehanat; Lotrakul, Pongtharin; Punnapayak, Hunsa
An extracellular ?-amylase from the obligate halophilic Aspergillus penicillioides TISTR3639 strain was produced and enriched to apparent homogeneity by ammonium sulfate precipitation and Sephadex G100 gel filtration column chromatography. The mass of the purified amylase was estimated to be 42?kDa by SDS-PAGE. With soluble starch as the substrate it had a specific activity of 118.42?U·mg?1 and Vmax? and Km values of 1.05?µmol·min?1·mg?1 and 5.41?mg·mL?1, respectively. The enzyme was found to have certain polyextremophilic characteristics, with an optimum activity at pH 9, 80°C, and 300?g·L?1 NaCl. The addition of CaCl2 at 2?mM was found to slightly enhance the amylase activity, while ZnCl2, FeCl2, or EDTA at 2?mM was strongly or moderately inhibitory, respectively, suggesting the requirement for a (non-Fe2+ or Zn2+) divalent cation. The enzyme retained more than 80% of its activity when incubated with three different laundry detergents and had a better performance compared to a commercial amylase and three detergents in the presence of increasing NaCl concentrations up to 300?g·L?1. Accordingly, it has a good potential for use as an ?-amylase in a low water activity (high salt concentration) and at high pH and temperatures. PMID:26180787
What's New in Salivary Gland Cancer Research and Treatment?
... Topic Additional resources for salivary gland cancer What's new in salivary gland cancer research and treatment? Medical ... they hope to use this information to develop new treatments that work better and cause fewer side ...
Comparisons of salivary proteins from five aphid (Hemiptera: Aphididae) species
Technology Transfer Automated Retrieval System (TEKTRAN)
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 importan...
Use of piezoelectric shock waves to effect release of amylase from capsules containing the amylase producing rat pancreatic tumour cell line AR42J
B. Jones; U. McCarthy; L. Bunni; L. McHale; M. Butler; J. Seary; A. P. McHale
Summary The amylase-producing rat pancreatic tumour cell line AR42J was encapsulated in alginate\\/poly-L-lysine under conditions where the secreted amylase was retained within the system. On treatment with shock waves from a lithotriptor, the porosity of the capsules was changed and the amylase was released from the system. Conditions of treatment were manipulated in order to maximise release of the amylase
A comparison of barley malt osmolyte concentrations and standard malt quality measurements as indicators of barley malt amylolytic enzyme activities
Technology Transfer Automated Retrieval System (TEKTRAN)
This study was conducted to test the hypothesis that barley malt osmolyte concentrations (OC) would correlate better with malt a-amylase, ß-amylase, and limit dextrinase activities than do the standard malt quality measurements (malt extract [ME], diastatic power [DP], ASBC a-amylase activity, solub...
Cloning, expression, and purification of insect (Sitophilus oryzae) alpha-amylase, able to digest granular starch, in Yarrowia lipolytica host.
Celi?ska, Ewelina; Bia?as, Wojciech; Borkowska, Monika; Grajek, W?odzimierz
Raw-starch-digesting enzymes (RSDE) are of major importance for industrial applications, as their usage greatly simplifies the starch processing pipeline. To date, only microbial RSDE have gained considerable attention, since only microbial production of enzymes meets industrial demands. In this study, ?-amylase from rice weevil (Sitophilus oryzae), the major rice pest, was cloned and expressed in Yarrowia lipolytica Po1g strain. The enzyme was secreted into the culture medium, and the peak activity (81 AU/L) was reached after only 29 h of culturing in 5-L bioreactors. Through simple purification procedure of ammonium sulfate precipitation and affinity chromatography, it was possible to purify the enzyme to apparent homogeneity (25-fold purification factor, at 5 % yield). The optimal conditions for the ?-amylase activity were pH 5.0 and a temperature of 40 °C. The ?-amylase studied here did not show any obligate requirement for Ca(2+) ions. The recombinant ?-amylase appeared to efficiently digest granular starch from pea, amaranth, waxy corn, and waxy rice. PMID:25547839
Heat, Acid and Chemically Induced Unfolding Pathways, Conformational Stability and Structure-Function Relationship in Wheat ?-Amylase
Singh, Kritika; Shandilya, Manish; Kundu, Suman; Kayastha, Arvind M.
Wheat ?-amylase, a multi-domain protein with immense industrial applications, belongs to ?+? class of proteins with native molecular mass of 32 kDa. In the present study, the pathways leading to denaturation and the relevant unfolded states of this multi-domain, robust enzyme from wheat were discerned under the influence of temperature, pH and chemical denaturants. The structural and functional aspects along with thermodynamic parameters for ?-amylase unfolding were probed and analyzed using fluorescence, circular dichroism and enzyme assay methods. The enzyme exhibited remarkable stability up to 70°C with tendency to aggregate at higher temperature. Acid induced unfolding was also incomplete with respect to the structural content of the enzyme. Strong ANS binding at pH 2.0 suggested the existence of a partially unfolded intermediate state. The enzyme was structurally and functionally stable in the pH range 4.0–9.0 with 88% recovery of hydrolytic activity. Careful examination of biophysical properties of intermediate states populated in urea and GdHCl induced denaturation suggests that ?-amylase unfolding undergoes irreversible and non-coincidental cooperative transitions, as opposed to previous reports of two-state unfolding. Our investigation highlights several structural features of the enzyme in relation to its catalytic activity. Since, ?-amylase has been comprehensively exploited for use in a range of starch-based industries, in addition to its physiological significance in plants and animals, knowledge regarding its stability and folding aspects will promote its biotechnological applications. PMID:26053142
Diagnosis of intestinal amebiasis using salivary IgA antibody detection.
del Muro, R; Acosta, E; Merino, E; Glender, W; Ortiz-Ortiz, L
This investigation sought to determine whether detection of salivary IgA antibodies to Entamoeba histolytica could identify intestinal amebic infections among 223 school children. Four groups of children were identified through coproparasitoscopic examination: E. histolytica as other parasites only (20%); and parasite-free (25%). The diagnostic accuracy of salivary IgA antibodies to an E. histolytica membrane extract was 91.5% (sensitivity, 85%; specificity, 98%), maintaining high predictive value at different prevalences. Also, a positive correlation (r = .753, P less than .001) was observed between fecal E. histolytica membrane antigen levels and salivary IgA antibody activity. Measurement of IgA antibodies in saliva may be useful in diagnosing intestinal infections with E. histolytica within a wide range of prevalences. Moreover, sampling of saliva may be a useful non invasive test for immunoepidemiologic surveys. PMID:2230266
A new method for the determination of ? -amylase
H. Rinderknecht; P. Wilding; B. J. Haverback
Zusammenfassung Es wird eine einfache und empfindliche Methode zur Bestimmung der ?-Amylase mit Hilfe eines durch Remazolbrillantblau R kovalent markierten Stärkesubstrates beschrieben. Die Bestimmung der Enzymwirkung erfolgt durch colorimetrische Messung der Spaltprodukte nach Entfernung des überschüssigen Substrates.
21 CFR 862.1070 - Amylase test system.
Code of Federal Regulations, 2010 CFR
...ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Chemistry Test Systems § 862.1070 Amylase test system. (a) Identification. An...
21 CFR 862.1070 - Amylase test system.
Code of Federal Regulations, 2011 CFR
...ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Chemistry Test Systems § 862.1070 Amylase test system. (a) Identification. An...
Salivary proteomics in biomedical research.
Zhang, Aihua; Sun, Hui; Wang, Ping; Wang, Xijun
Proteins that are important indicators of physiological or pathological states, can provide information for the identification of early and differential markers for disease. Saliva, contains an abundance of proteins, offers an easy, inexpensive, safe, and non-invasive approach for disease detection, and possesses a high potential to revolutionize the diagnostics. Discovery of salivary biomarkers could be used to scrutinize health and disease surveillance. The impact of human saliva proteome analysis in the search for clinically relevant disease biomarkers will be realized through advances made using proteomic technologies. The advancements of emerging proteomic techniques have benefited biomarker research to the point where saliva is now recognized as an excellent diagnostic medium for the detection of disease. This review presents an overview of the value of saliva as a credible diagnostic tool and we aim to summarize the proteomic technologies currently used for global analysis of saliva proteins and to elaborate on the application of saliva proteomics to the discovery of disease biomarkers, and discuss some of the critical challenges and perspectives in this field. PMID:23146870
Distribution and evolution of introns in drosophila amylase genes
Jean-Luc Da Lage; Maurice Wegnez; Marie-Louise Cariou I
While the two amylase genes ofDrosophila melanogaster are intronless, the three genes ofD. pseudoobscura harbor a short intron. This raises the question of the common structure of theAmy gene in Drosophila species. We have investigated the presence or absence of an intron in the amylase genes of 150 species\\u000a of Drosophilids. Using polymerase chain reaction (PCR), we have amplified a
RNA complementary to ?-amylase mRNA in barley
John C. Rogers
Two experimental approaches demonstrate that different types of RNA complementary to a-amylase mRNA are present in barley. S1 nuclease assays identify an RNA that is complementary to essentially the full length of both the type A and type B a-amylase mRNAs. Complementarity, however, is imperfect: the S1 nuclease-resistant products can only be identified if they are electrophoresed as RNA-DNA hybrids.
Early morning salivary cortisol is not associated with extraversion
Early morning salivary cortisol is not associated with extraversion Marcus R. Munafo` a,*, Laura salivary cortisol levels and neuroticism has recently been reported, but it is unclear whether early morning salivary cortisol levels in individuals selected for high and low extraversion. Thirty
Saliva and the Salivary Glands in the Elderly
Louis Mandel
Salivary secretory dysfunction and\\/or salivary gland pathology are conditions that can be found in all segments of the population. The aged are uniquely susceptible to some of these entities which may reflect manifestations of local, systemic or even imagined disorders. Many other salivary gland disease processes, both local and systemic, do not particularly seek out the elderly nor do they