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1

Stress-induced changes in human salivary alpha-amylase activity -- associations with adrenergic activity.  

PubMed

The salivary enzyme alpha-amylase has been proposed to indicate stress-reactive bodily changes. A previous study by the authors revealed marked increases in salivary alpha-amylase following psychosocial stress, indicating a stress-dependent activation of salivary alpha-amylase. Salivary alpha-amylase has been suggested to reflect catecholaminergic reactivity. Our aim was to assess/evaluate a possible relationship between salivary alpha-amylase and adrenergic parameters, i.e. catecholamines, as well as other stress markers. Using an intra-individual repeated measures design, 30 healthy young men underwent the Trier Social Stress Test (TSST), which consists of a mental arithmetic task and free speech in front of an audience and a control condition in randomized order. Salivary alpha-amylase and salivary cortisol as well as plasma catecholamines and cardiovascular activity were repeatedly measured before, during, and after both conditions. Significant differences were found between the stress and the rest condition in salivary alpha-amylase, salivary cortisol, plasma catecholamines, and cardiovascular parameters (heart rate, LF, HF, LF/HF). However, general alpha-amylase responses (area under the curve) were not associated with general responses in catecholamines and cortisol in the stress condition (r smaller than 0.25 for all analyses). Analysis of cardiovascular parameters indicates a positive relationship between amylase and sympathetic tone (LF/HF) during stress. Salivary alpha-amylase is sensitive to psychosocial stress. Since it does not seem to be closely related to other biological stress markers such as catecholamines and cortisol, salivary alpha-amylase may be a useful additional parameter for the measurement of stress. PMID:16002223

Nater, Urs Markus; La Marca, Roberto; Florin, Ladina; Moses, Anthony; Langhans, Wolfgang; Koller, Markus M; Ehlert, Ulrike

2006-01-01

2

Clinical Performance of a Salivary Amylase Activity Monitor During Hemodialysis Treatment  

PubMed Central

The hemodialysis procedure is thought to be a physical stressor in the majority of hemodialyzed patients. Previous studies suggest that elevated salivary amylase level may correlate with increased plasma norepinephrine level under psychological and physical stress conditions. In this study, we investigated biological stress reactivity during hemodialysis treatment using salivary amylase activity as a biomarker. Seven patients (male/female = 5/2, age: 67.7+/?5.9 years) who had been receiving regular 4 h hemodialysis were recruited. Salivary amylase activity was measured using a portable analyzer every hour during the hemodialysis session. Salivary amylase activity was shown to be relatively stable and constant throughout hemodialysis, whereas there were significant changes in systolic blood pressure and pulse rate associated with blood volume reduction. Our results show that hemodialysis treatment per se dose not affect salivary amylase activity.

Shimazaki, Masaru; Matsuki, Takayuki; Yamauchi, Kazuaki; Iwata, Michihiro; Takahashi, Hiroshi; Sakamoto, Kenichi; Ohata, Junichi; Nakamura, Yuichi; Okazaki, Yusuke

2008-01-01

3

Stress affects salivary alpha-Amylase activity in bonobos.  

PubMed

Salivary alpha-Amylase (sAA) is a starch digesting enzyme. In addition to its function in the context of nutrition, sAA has also turned out to be useful for monitoring sympathetic nervous system activity. Recent studies on humans have found a relationship between intra-individual changes in sAA activity and physical and psychological stress. In studies on primates and other vertebrates, non-invasive monitoring of short-term stress responses is usually based on measurements of cortisol levels, which are indicative of hypothalamic-pituitary-adrenal activity. The few studies that have used both cortisol levels and sAA activity indicate that these two markers may respond differently and independently to different types of stress such that variation in the degree of the activation of different stress response systems might reflect alternative coping mechanisms or individual traits. Here, we present the first data on intra- and inter-individual variation of sAA activity in captive bonobos and compare the results with information from other ape species and humans. Our results indicate that sAA activity in the bonobo samples was significantly lower than in the human samples but within the range of other great ape species. In addition, sAA activity was significantly higher in samples collected at times when subjects had been exposed to stressors (judged by changes in behavioral patterns and cortisol levels) than in samples collected at other times. Our results indicate that bonobos possess functioning sAA and, as in other species, sAA activity is influenced by autonomic nervous system activity. Monitoring sAA activity could therefore be a useful tool for evaluating stress in bonobos. PMID:21945369

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

2012-01-18

4

Determinants of the diurnal course of salivary alpha-amylase  

Microsoft Academic Search

Summary Objective: Previous data from our group and others have shown that salivary alpha- amylase activity increases in response to stress. It has been suggested that salivary alpha- amylase may be a marker for adrenergic activity. Less is known about other determinants of salivary alpha-amylase activation. The objective of the current study was to describe the diurnal pattern of salivary

Urs M. Natera; Nicolas Rohleder; Wolff Schlotze; Ulrike Ehlert; Clemens Kirschbaum

5

Psychosocial stress-induced activation of salivary alpha-amylase: an indicator of sympathetic activity?  

PubMed

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 cortisol measurements, in which noninvasive samples can be taken at any place and stored at room temperature for sufficient time before later analysis in the laboratory. Salivary alpha-amylase (sAA) is a candidate that may prove useful in this context. We show here that sAA activity is increased by acute psychosocial stress (Trier Social Stress Test) and that increases in sAA correlate with increases in norepinephrine. We further report that sAA exhibits a stable circadian pattern that mirrors that of salivary cortisol. In conclusion, the current data show that salivary alpha-amylase may serve as an easy-to-use index for SAM activity. However, some questions remain to be answered; for example, what impact does salivary flow rate exert on stress-induced sAA activity? PMID:15677423

Rohleder, Nicolas; Nater, Urs M; Wolf, Jutta M; Ehlert, Ulrike; Kirschbaum, Clemens

2004-12-01

6

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

ERIC Educational Resources Information Center

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

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

2009-01-01

7

Salivary Alpha-Amylase Activity and Salivary Flow Rate in Young Adults  

PubMed Central

The secretion of salivary alpha-amylase (sAA) is more associated with psychoneuroendocrinological response to stress than with the flow rate and age. The aim of this cross sectional study is to build an explanatory model based on patterns of relationship between age 20-39 in resting and stimulated saliva under no stressful condition in healthy volunteers. Both resting and stimulated saliva were collected from 40 subjects. The sAA values were log-transformed, the normality assumption was verified with the Shapiro-Wilk test and the reliability of the measurements was estimated by the Pearsons’ r correlation coefficient. The estimated model was based on the theory of the Linear Mixed Models. Significant mean changes were observed in flow rate and sAA activity between resting and stimulated saliva. The final model consists of two components, the first revealed a positive correlation between age and sAA while the second one revealed a negative correlation between the interaction of age × flow rate in its condition (resting or stimulated saliva), with sAA. Both flow rate and age influence sAA activity.

Arhakis, Aristidis; Karagiannis, Vasilis; Kalfas, Sotirios

2013-01-01

8

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

PubMed Central

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

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

2013-01-01

9

Development of a low cost self-sensing Piezoresistive Microcantilever biosensor and read-out circuitry for measuring salivary-amylase activity  

Microsoft Academic Search

This paper deals with the development of a low cost self-sensing Piezeoresistive Microcantilever Biosensor to detect human stress using salivary alpha amylase activity. The salivary alpha amylase enzyme will immobilize on to the Piezeoresistive Microcantilever biosensor to detect activity during the enzyme responses. When the Microcantilever beam deflects it caused the stress change within the microcantilever beam and applied strain

Nina Korlina Madzhi; Lee Yoot Khuan; M. F. Abdullah; A. Ahmad

2010-01-01

10

Substrate differentiation of human pancreatic and salivary alpha-amylases  

Microsoft Academic Search

To increase specificity in the clinical determination of amylase, the differential activities of human pancreatic and salivary amylases toward two different substrates—a Lintner soluble starch (LSS) and an insoluble, dyed starch, Amylose Azure (AA)—were investigated. A ratio of initial-velocity activities of Lintner soluble starch to Amylose Azure (LSS\\/AA) for salivary amylase was at least twice as great as the LSS\\/AA

F. F. Hall; C. R. Ratliff; T. Hayakawa; T. W. Culp; N. C. Hightower

1970-01-01

11

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

PubMed Central

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

2014-01-01

12

Salivary Alpha-Amylase Activity: A Possible Indicator of Pain-Induced Stress in Orthodontic Patients  

Microsoft Academic Search

IntroductionPain, a common experience reported by orthodontic patients, has its intensity assessed with the help of subjective scales, which have a limited and disputable value. Such unpleasant experience, which may raise stress levels, is reflected by an increase in the salivary concentration of alpha-amylase.

Marcio José da Silva Campos; Nádia Rezende Barbosa Raposo; Ana Paula Ferreira; Robert Willer Farinazzo Vitral

2011-01-01

13

Oral Fusobacterium nucleatum subsp. polymorphum binds to human salivary ?-amylase.  

PubMed

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

Zulfiqar, M; Yamaguchi, T; Sato, S; Oho, T

2013-12-01

14

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

PubMed

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

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

2014-07-01

15

Elevated serum amylase activity in the absence of clinical pancreatic or salivary gland disease: possible role of acute hypoxemia.  

PubMed

Elevated serum amylase activity, in the absence of clinically apparent pancreatic or salivary gland disease, has been observed in many seemingly unrelated conditions. In a search for common etiological factors to account for hyperamylasemia in these conditions, a retrospective analysis was performed. Eighty-four episodes of hyperamylasemia (greater than 300 I.U./l. Phadebas method) occurring in 75 patients over a one-year period ending in June, 1975 were assigned to one of two groups. Group 1 consisted of 56 (67%) episodes of hyperamylasemia with clinical pancreatitis. Group 2 consisted of 28 (33%) episodes of hyperamylasemia in the absence of clinical pancreatitis. Hypoxemia (pO2 less than 75 mm. Hg.) was found in 9/15 patients in Group 2 who had arterial blood gases measured. To assess the possible relationship between acute hypoxemia and amylase activity, a prospective study was initiated. Patients with known causes of pancreatitis or renal failure were eliminated. Hyperamylasemia was found in 3/8 hypoxemic patients. This raises the possibility that acute hypoxemia alone or in combination with other factors may raise serum amylase activity, possibly through ischemic injury to the pancreas or salivary glands or other amylase containing tissues. PMID:742604

Jam, I; Shoham, M; Wolf, R O; Mishkin, S

1978-11-01

16

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

Microsoft Academic Search

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

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

2004-01-01

17

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

PubMed

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

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

2014-08-01

18

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

PubMed Central

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

Mandel, Abigail L.

2012-01-01

19

Enhancement of cyclic AMP modulated salivary amylase secretion by protein kinase C activators.  

PubMed

The effect of protein kinase C activators on isoproterenol-induced amylase secretion were investigated in isolated rat parotid cells. Pretreatment with phorbol dibutyrate potentiated isoproterenol-induced amylase secretion. This effect of phorbol dibutyrate was mimicked by dioctanoylglycerol or carbachol. Phorbol dibutyrate also potentiated secretion evoked by the adenylate cyclase activator forskolin and by dibutyryl cAMP. Neither phorbol dibutyrate nor carbachol enhanced isoproterenol-induced cAMP accumulation. The present study reveals a coordinate interaction between cAMP and protein kinase C at a step in the secretory mechanism distal to cAMP generation. PMID:2462421

McKinney, J S; Rubin, R P

1988-12-01

20

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

PubMed Central

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

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

2014-01-01

21

Characterization of salivary alpha-amylase binding to Streptococcus sanguis.  

PubMed Central

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

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

1989-01-01

22

Characterization of salivary alpha-amylase binding to Streptococcus sanguis  

SciTech Connect

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

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

1989-09-01

23

Genetic Variation in Amount of Salivary Amylase in the Bank Vole, CLETHRIONOMYS GLAREOLA  

PubMed Central

Several investigated bank vole populations are polymorphic for the number of salivary amylase loci, and individual chromosomes may carry one, two or three linked amylase structural genes. In the present study, we have used bank vole stocks homozygous for different chromosomes to investigate the relationship between amylase production and gene number. By measuring the amylase activity in parotid glands and the percentage of amylase protein in saliva, we have been able to demonstrate that the amount of salivary amylase is directly proportional to the proposed gene number. The paper also describes the allele, AmySu, which codes for a heat-labile salivary amylase. The relative amounts of the heat-labile isozyme have been determined in different heterozygotes containing this allele, and these results also support the multiple locus model. Finally, a stock devoid of salivary amylase activity was established. Animals from this strain have, however, a protein in the parotid glands and in saliva that is very similar to amylase in molecular weight, amino acid composition and in its binding to glycogen and cyclohepta-amylose. In genetic crosses, the protein segregates as an amylase allele. Therefore, this protein, encoded by the functionally null allele AmyN, may represent an incorrectly processed amylase precursor.

Hjorth, J. Peter; Meisler, Miriam; Nielsen, J. T?nnes

1979-01-01

24

Salivary amylase activity is useful for assessing perioperative stress in response to pain in patients undergoing endoscopic submucosal dissection of gastric tumors under deep sedation  

Microsoft Academic Search

Background  Although endoscopic submucosal dissection (ESD) for patients with gastric tumors under the conditions of unconsciousness is\\u000a considered to be minimally invasive, no objective assessment of the perioperative stress of ESD has yet been conducted. Today,\\u000a stress levels can be easily and objectively assessed by monitoring salivary amylase activity (sAMY). We evaluated the perioperative\\u000a changes in the sAMY in patients undergoing

Masaya Uesato; Yoshihiro Nabeya; Takashi Akai; Masahito Inoue; Yoshiyuki Watanabe; Hiroshi Kawahira; Toshitaka Mamiya; Yoshihito Ohta; Ryuji Motojima; Akiko Kagaya; Yorihiko Muto; Hideki Hayashi; Hisahiro Matsubara

2010-01-01

25

Variations of salivary amylase as a test of internal irradiation in man.  

PubMed

The hypothesis of the parotid gland radiosensitivity has suggested to the authors to follow up the effect of therapeutic internal irradiation on the serum, urine and salivary alpha-amylase activity in man. The assay and visualization of alpha-amylase isoenzymes on agar gel zymograms by using the "Phadebas amylase test" tablets has demonstrated in some cases increases of salivary alpha-amylase activity expressed by the ratio of the optical density after internal irradiation to that before administration of radioisotopes. PMID:1243186

Zamfirescu-Gheorghiu, M; Cheta, N; Funduc, I; Ciobanu, F

1975-01-01

26

Individual Differences in AMY1 Gene Copy Number, Salivary ?-Amylase Levels, and the Perception of Oral Starch  

PubMed Central

Background The digestion of dietary starch in humans is initiated by salivary ?-amylase, an endo-enzyme that hydrolyzes starch into maltose, maltotriose and larger oligosaccharides. Salivary amylase accounts for 40 to 50% of protein in human saliva and rapidly alters the physical properties of starch. Importantly, the quantity and enzymatic activity of salivary amylase show significant individual variation. However, linking variation in salivary amylase levels with the oral perception of starch has proven difficult. Furthermore, the relationship between copy number variations (CNVs) in the AMY1 gene, which influence salivary amylase levels, and starch viscosity perception has not been explored. Principal Findings Here we demonstrate that saliva containing high levels of amylase has sufficient activity to rapidly hydrolyze a viscous starch solution in vitro. Furthermore, we show with time-intensity ratings, which track the digestion of starch during oral manipulation, that individuals with high amylase levels report faster and more significant decreases in perceived starch viscosity than people with low salivary amylase levels. Finally, we demonstrate that AMY1 CNVs predict an individual's amount and activity of salivary amylase and thereby, ultimately determine their perceived rate of oral starch viscosity thinning. Conclusions By linking genetic variation and its consequent salivary enzymatic differences to the perceptual sequellae of these variations, we show that AMY1 copy number relates to salivary amylase concentration and enzymatic activity level, which, in turn, account for individual variation in the oral perception of starch viscosity. The profound individual differences in salivary amylase levels and salivary activity may contribute significantly to individual differences in dietary starch intake and, consequently, to overall nutritional status.

Mandel, Abigail L.; Peyrot des Gachons, Catherine; Plank, Kimberly L.; Alarcon, Suzanne; Breslin, Paul A. S.

2010-01-01

27

Effect of an herb root extract, herbal dentifrice and synthetic dentifrice on human salivary amylase  

PubMed Central

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.

Sapra, Gaurav; Vyas, Yogesh Kumar; Agarwal, Rahul; Aggarwal, Ashish; Chandrashekar, K T; Sharma, Kanika

2013-01-01

28

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

PubMed Central

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

Sadi, Hana; Finkelman, Matthew; Rosenberg, Morton

2013-01-01

29

Salivary alpha-amylase and cortisol responsiveness following electrical stimulation stress in major depressive disorder patients  

Microsoft Academic Search

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

Yoshihiro Tanaka; Yoshinobu Ishitobi; Yoshihiro Maruyama; Aimi Kawano; Tomoko Ando; Shizuko Okamoto; Masayuki Kanehisa; Haruka Higuma; Taiga Ninomiya; Jusen Tsuru; Hiroaki Hanada; Kensuke Kodama; Koichi Isogawa; Jotaro Akiyoshi

30

Immunological relationships and post-translational modifications of human salivary amylase (Amy 1 ) and pancreatic amylase (Amy 2 ) isozymes  

Microsoft Academic Search

Electrophoretic phenotypes of human salivary amylase (Amy1) and pancreatic amylase (Amy2) consist of complex isozyme patterns which may result from post-translational modifications of the primary products of the amylase loci. Biochemical separation of the two molecular weight families of salivary amylase and development of a new electrophoretic system have allowed the identification of complete isozyme patterns corresponding to variant alleles

R. C. Karn; Barnett B. Rosenblum; Jewell C. Ward; A. Donald Merritt; Jeff D. Shulkin

1974-01-01

31

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

PubMed

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

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

2012-06-14

32

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

PubMed Central

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

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

2014-01-01

33

Flow injection spectrophotometric analysis of human salivary ?-amylase activity using an enzyme degradation of starch-iodine complexes in flow channel and its application to human stress testing.  

PubMed

Flow injection spectrophotometric analysis (FIA) of human salivary ?-amylase was developed using an enzyme degradation reaction of starch-iodine complexes. In this proposed method, the salivary ?-amylase, known as a human stress indicator, is directly and rapidly determined without any pretreatment. In this study, the optimum starch-iodine complexes (i.e., optimum molecular weight and amylase-amylopectin compounding ratio) were selected, and their rapid degradation in the flow channel was investigated to determine salivary amylase in the FIA system. The determination range of ?-amylase was obtained from 0.25 to 5.0 kilo Novo unit per milliliter (KNU/mL), and these concentrations were equivalent to the real concentration of amylase in human saliva. The quantitative values obtained by this method were found to be highly reproducible with 1.6% (n=25) of the relative standard deviation for 1.0?KNU/mL. The detection limit (3?) was 60 NU/mL. In addition, the method requires small volume of a sample (20?µL), and 30 samples was sequentially measured within one hour. Real human saliva collected before and after exercise was utilized to demonstrate the feasibility of human stress test and analytical performance of this approach. PMID:24189429

Ohtomo, Takao; Igarashi, Shukuro; Takagai, Yoshitaka

2013-01-01

34

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

Microsoft Academic Search

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

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

2008-01-01

35

Relation between salivary amylase and cortisol respnses to different stress tasks: impact of sex  

Microsoft Academic Search

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

Stegeren van A. H; O. T. Wolf; M. Kindt

2008-01-01

36

Aging diurnal rhythms and chronic stress: Distinct alteration of diurnal rhythmicity of salivary ?-amylase and cortisol  

Microsoft Academic Search

The present study assessed diurnal profiles of salivary ?-amylase (sAA), proposed as a marker of autonomic activity, and salivary cortisol in competitive ballroom dancers as well as age- and sex-matched controls to investigate age-related changes of basal activity and potential chronic psychosocial stress-related alterations. According to the Allostatic Load (AL) hypothesis of a cumulative wear and tear of the body

Jana Strahler; Christiane Berndt; Clemens Kirschbaum; Nicolas Rohleder

2010-01-01

37

Salivary Amylase Promotes Adhesion of Oral Streptococci to Hydroxyapatite  

Microsoft Academic Search

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

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

1995-01-01

38

Human salivary alpha-amylase reactivity in a psychosocial stress paradigm  

Microsoft Academic Search

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

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

2005-01-01

39

Salivary Alpha Amylase-Cortisol Asymmetry in Maltreated Youth  

PubMed Central

Background Maltreatment represents a major stressor in the lives of many youth. Given the known effects of stress exposure on subsequent biological stress response systems, researchers have been interested in the effects of maltreatment on the functioning of these systems. Experimental studies reveal that previous exposure to stress affects the symmetry between components of the physiological stress response to subsequent stress. The present study examined asymmetry between salivary alpha amylase (sAA), a sympathetic indicator, and cortisol reactivity to a social stressor among maltreated and comparison youth age 9 to 14 years. Consistent with earlier studies suggesting that stress leads to asymmetry between hypothalamic-pituitary-adrenal axis and sympathetic nervous system activity, we expected that maltreated youth would exhibit greater sAA-cortisol asymmetry than would comparison youth. Methods Forty-seven maltreated and 37 comparison youth visited the lab and engaged in a social stress protocol. We collected 2 saliva samples before the stressor and 4 after, at 0 minutes post stress and every 10 minutes for 30 minutes. Results Maltreatment status moderated the relation between sAA and cortisol activity in response to the stressor. Comparison youth showed significant links between the sAA and cortisol responses; maltreated youth had no significant associations between responses in the two biomarkers. Conclusion The data were consistent with sAA-cortisol asymmetry among maltreated youth. Further research should seek to replicate this finding and investigate its implication for developmental trajectories.

Gordis, Elana B.; Granger, Douglas A.; Susman, Elizabeth J.; Trickett, Penelope K.

2008-01-01

40

Interparental Aggression and Parent-Adolescent Salivary Alpha Amylase Symmetry  

PubMed Central

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

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

2010-01-01

41

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

Microsoft Academic Search

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

Jutta M. Wolf; Erin Nicholls; Edith Chen

2008-01-01

42

Diurnal trajectories of salivary cortisol, salivary alpha-amylase and psychological profiles in oral lichen planus patients.  

PubMed

Although many reports have been published on the link between oral lichen planus (OLP) and the stress-related neuro-psycho-endocrine clinical features of the disease over the last 20 years, the data still remain controversial. Therefore, the aim of this study was to explore the personality traits of OLP subjects and assess the subjects? capability of coping with stress challenges. Cortisol and alpha-amylase were measured as reliable markers of the hypothalamic-pituitary-adrenal (HPA) axis and autonomic nervous system (ANS) activities in salivary samples collected by the participants at their home during the sampling day (07:30, 12:00, and 19:30). Compared with the healthy controls, the OLP patients demonstrated a less effective coping ability, had higher scores in stress perception and loneliness, and had no significant variation in their anxiety and depressive symptoms. The OLP patients also showed dysregulation of the HPA axis activity with a significant reduction of diurnal salivary cortisol production, which was particularly significant in the morning hours. No significant variation was found in the OLP salivary alpha-amylase diurnal fluctuation and production, which was measured at the same time point as that for cortisol. In conclusion, we report that OLP subjects had a reduced capability of coping with stress events and presented a dysregulation of HPA axis activity with hypocortisolism detected in the morning hours. PMID:24750801

Pippi, R; Patini, R; Ghiciuc, C M; Sandu, R B; Pasquali, V; Scaccianoce, S; Dima-Cozma, L C; Patacchioli, F R

2014-01-01

43

Renal clearance of pancreatic and salivary amylase relative to creatinine in patients with chronic renal insufficiency.  

PubMed Central

Pancreatic and salivary amylase/creatinine clearance ratios in patients with various degrees of renal impairment were compared with those obtained for control subjects. In chronic renal insufficiency (mean GFR 30 ml/min +/- 15 SD; n = 13) the clearance ratios for pancreatic (mean 3.5 +/- 1.85 SD) and salivary (mean 2.3 +/- 1.3 SD) amylase were significantly higher (P less than 0.05) than those in controls. Corresponding control values (n = 26) were 2.64 +/- 0.86 (pancreatic) and 1.64 +/- 0.95 (salivary). Three patients showed values above the normal limit. In the diabetic group (mean GFR 41 ml/min +/- 22 SD; n = 10) salivary amylase/creatinine clearance ratios (mean 2.36 +/- 1.55 SD) were significantly higher than in controls (P less than 0.05). Three patients showed raised values. Pancreatic amylase clearance was raised in only one of these patients. Three patients with terminal disease (mean GFR 10 ml/min) showed markedly raised (two- to threefold) clearance ratios for both salivary and pancreatic amylase. Of a total of 26 patients, eight had increased total amylase/creatinine clearance ratios. Pancreatic amylase/creatinine clearance was increased in seven patients, while nine patients showed raised salivary amylase/creatinine ratios. Patients with raised clearance ratios did not have clinical evidence of pancreatitis. We suggest that, in the presence of impaired renal function, a high amylase/creatinine clearance ratio need not be indicative of pancreatic disease.

Keogh, J B; McGeeney, K F; Drury, M I; Counihan, T B; O'Donnell, M D

1978-01-01

44

Salivary Secretion of Rat Haptocorrin and Amylase Is Stimulated by Vasoactive Intestinal Polypeptide  

Microsoft Academic Search

Rat salivary glands secrete the cobalamin-binding protein haptocorrin and the enzyme amylase. We report the secretion to be stimulated in a dose-dependent manner by vasoactive intestinal polypeptide (VIP). VIP especially enhances the secretion of amylase. The concentration of amylase increases in saliva and in serum while the concentration of haptocorrin increases only in saliva. Serum contains a cobalamin-binding protein that

E. Nexř; M. Hansen; Skov Olsen; E. Magid

1987-01-01

45

Alpha-amylase is a human salivary protein with affinity to lipopolysaccharide of Aggregatibacter actinomycetemcomitans.  

PubMed

Aggregatibacter actinomycetemcomitans lipopolysaccharide (Aa.LPS) is a major virulence factor associated with aggressive periodontitis. Although the recognition of Aa.LPS is potentially initiated by salivary proteins in the oral cavity, Aa.LPS-binding proteins (Aa.LPS-BPs) in saliva are poorly characterized. The purpose of this study was to capture and identify Aa.LPS-BPs in human saliva using a LTQ-Orbitrap hybrid Fourier transform mass spectrometry. Aa.LPS conjugated onto N-hydroxysuccinimidyl-Sepharose(®) 4 Fast Flow beads (Aa.LPS-beads) activated Toll-like receptor 4 and produced nitric oxide and Interferon gamma-inducible protein-10, implying that the conjugation process did not alter the biological properties of Aa.LPS. Aa.LPS-BPs were subsequently isolated from the nine human saliva samples from healthy individuals with the Aa.LPS-beads followed by identification with the mass spectrometry. Aa.LPS-BPs include ?-amylase, serum albumin, cystatin, lysozyme C, submaxillary gland androgen-regulated protein 3B, immunoglobulin subunits, polymeric immunoglobulin receptor, deleted in malignant brain tumors 1, prolactin-inducible protein, lipocalin-1, and basic salivary proline-rich protein 2. Specific binding was validated using a pull-down assay with ?-amylase which was captured at the highest frequency. Alpha-amylase demonstrated to interfere with the adherence and biofilm formation of A. actinomycetemcomitans. Even heat-inactivated ?-amylase showed the interference to the same extent. Conclusively, we identified unique Aa.LPS-BPs that provide useful information to understand bacterial pathogenesis and host innate immunity in the oral cavity. PMID:23194029

Baik, J E; Hong, S W; Choi, S; Jeon, J H; Park, O-J; Cho, K; Seo, D-G; Kum, K-Y; Yun, C-H; Han, S H

2013-04-01

46

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

PubMed Central

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

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

2013-01-01

47

Salivary ?-amylase and cortisol responsiveness following electrical stimulation stress in panic disorder patients.  

PubMed

Psychosocial stress-induced activation of salivary ?-amylase (sAA) functions is as a marker of sympathoadrenal medullary system (SAM) activity. However, in contrast to salivary cortisol, sAA has been less extensively studied in panic disorder patients. The present study measured sAA and salivary cortisol levels in patients with panic disorder following electrical stimulation stress. The authors determined Profile of Mood State (POMS) scores and State-Trait anxiety Inventory (STAI) scores, heart rate variability (HRV), and levels of sAA and salivary cortisol in 34 patients with panic disorder and 41 healthy volunteers following the application of electrical stimulation stress. 34 alprazolam-treated patients with panic disorder were divided into non-responder and responder group. Vigor scores in patients with panic disorder were significantly decreased compared with healthy controls. Another score in POMS in patients with panic disorder were significantly increased compared with healthy controls. Trait and state anxiety of STAI in panic disorder patients were higher than healthy controls. There was no difference in either HRV or threshold of electrical stimulation applied between panic disorder patients and healthy controls. SAA levels in the responder group were significantly elevated compared with the non-responder group and controls both before and after electrical stimulation. In addition, there were no differences in salivary cortisol levels between responder and non-responder groups of patients with panic disorder and control. The sample may not be representative of the general population. These preliminary results suggest that sAA might be useful predictive biological markers of treatment responsiveness in patients with panic disorder. PMID:22391145

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

2012-05-01

48

Salivary alpha-amylase and cortisol responsiveness following electrical stimulation stress in major depressive disorder patients.  

PubMed

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

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

2012-03-30

49

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

Microsoft Academic Search

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

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

2006-01-01

50

Corrected sequences of cDNAs for human salivary and pancreatic alpha-amylases [corrected].  

PubMed

The nucleotide sequences of the cloned human salivary and pancreatic alpha-amylase cDNAs correspond to the continuous mRNA sequences of 1768 and 1566 nucleotides, respectively. These include all of the amino acid coding regions. Salivary cDNA contains 200 bp in the 5'-noncoding region and 32 in the 3'-noncoding region. Pancreatic cDNA contains 3 and 27 bp of 5'- and 3'-noncoding regions, respectively. The nucleotide sequence homology of the two cDNAs is 96% in the coding region, and the predicted amino acid sequences are 94% homologous. Comparison of the sequences of human alpha-amylase cDNAs with those previously obtained for mouse alpha-amylase genes (Hagenbuchle et al., 1980; Schibler et al., 1982) showed the possibility of gene conversion between the two genes of human alpha-amylase. PMID:6610603

Nishide, T; Emi, M; Nakamura, Y; Matsubara, K

1984-05-01

51

Oral intercourse or secondary transfer? A Bayesian approach of salivary amylase and foreign DNA findings.  

PubMed

The Bayesian Approach allows forensic scientists to evaluate the significance of scientific evidence in light of two conflicting hypothesis. This aids the investigator to calculate a numerical value of the probability that the scientific findings support one hypothesis over conflicting opinions. In the case where oral intercourse is alleged, ?-amylase, an indicator of saliva, is detected on penile swabs. The value of this finding is unknown as it may indicate the presence of saliva resulting from oral intercourse however it may also represent the presence of saliva due to innocent means such as background levels of salivary-?-amylase in the male population due to secondary transfer. Therefore, it is difficult to attach significance to this finding without background information and knowledge. A population study of the background levels of salivary-?-amylase was performed by analysing items of underwear worn under normal circumstances by 69 male volunteers. The Phadebas press test was used to screen the garments for amylase-containing stains and the positive areas were subjected to further confirmation of saliva by the RSID-Saliva kit. 44% of underwear screened had stains containing amylase. This study determined the background level of salivary-?-amylase and DNA on the inside front of male underwear which has potential implications on the interpretation of evidence in alleged oral intercourse. PMID:23683908

Breathnach, Michelle; Moore, Elizabeth

2013-06-10

52

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

ERIC Educational Resources Information Center

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

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

2012-01-01

53

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

ERIC Educational Resources Information Center

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

Marini, Isabella

2005-01-01

54

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

ERIC Educational Resources Information Center

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

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

2010-01-01

55

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

PubMed

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

Carboni, Ilaria; Rapi, Stefano; Ricci, Ugo

2014-07-01

56

Interaction of Salivary alpha-Amylase and Amylase-Binding-Protein A (AbpA) of Streptococcus gordonii with Glucosyltransferase of S. gordonii and Streptococcus mutans  

Microsoft Academic Search

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),

Biswendu Chaudhuri; Jennifer Rojek; M Margaret Vickerman; Jason M Tanzer; Frank A Scannapieco

2007-01-01

57

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

ERIC Educational Resources Information Center

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

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

2012-01-01

58

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

PubMed

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 Social Stress Test and a control condition. Saliva was sampled by passive drooling or salivettes. Saliva flow rate, sAA levels and output, salivary cortisol, and heart rate variability were measured. Flow rate increased only when sampled by passive drooling. Stress-induced increases in amylase levels were correlated with increases of amylase output but not with flow rate. Results indicate that flow rate is not a confounder of stress-induced sAA activation and suggest that valid measurements of sAA can be obtained by salivettes without the need for assessment of flow rate. PMID:17076822

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

2006-11-01

59

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

PubMed Central

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

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

2013-01-01

60

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

SciTech Connect

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

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

1988-09-12

61

A rapid method for separation and detection of human salivary amylase isoenzymes by isoelectric focusing in polyacrylamide gel.  

PubMed

Human salivary proteins were separated by isoelectric focusing on polyacrylamide gel in flat beds at 1000 V for 40 min. Amylase activity was detected after immersing the gel in 0.4 M tris-HCI buffer pH 7.4 to equilibrate the pH gradient. The enzyme activity was detected after diffusion into an overlayer of agarose gel containing an insoluble dye-starch polymer (Phadebas). Both whole human saliva and parotid saliva from 15 different persons contained four amylase components, except in three cases where only three bands were detected. The bands were all focused within a rather narrow pH range (pH 5.4-7.2) and the results were very reproducible. PMID:1065950

Wadström, T; Nord, C E; Kjellgren, M

1976-07-01

62

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

PubMed Central

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

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

2012-01-01

63

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

Microsoft Academic Search

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

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

64

Normative references of heart rate variability and salivary alpha-amylase in a healthy young male population  

PubMed Central

Background This study aimed to present normative reference values of heart rate variability and salivary alpha-amylase in a healthy young male population with a particular focus on their distribution and reproducibility. Methods The short-term heart rate variability of 417 young healthy Japanese men was studied. Furthermore, salivary alpha-amylase was measured in 430 men. The average age of the subjects were 21.9 years with standard deviation of 1.6 years. Interindividual variations in heart rate variability indices and salivary alpha-amylase levels were plotted as histograms. Data are presented as the mean, median, standard deviation, coefficient of variation, skewness, kurtosis, and fifth and 95th percentiles of each physiological index. Results Mean recorded values were heart period 945.85 ms, log-transformed high frequency component 9.84 ln-ms2, log-transformed low frequency component 10.42 ln-ms2, log-transformed low frequency to high frequency ratio 0.58 ln-ratio, standard deviation of beat-to-beat interval 27.17 ms and root mean square of successive difference 37.49 ms. The mean value of raw salivary alpha-amylase was 17.48 U/mL, square root salivary alpha-amylase 3.96 sqrt[U/mL] and log-transformed salivary alpha-amylase 2.65 ln[U/mL]. Log-transformed heart rate variability indices exhibited almost symmetrical distributions; however, time-domain indices of heart rate variability (standard deviation of beat-to-beat interval and root mean square of successive difference) exhibited right-skewed (positive skewness) distributions. A considerable right-skewed distribution was observed for raw salivary alpha-amylase. Logarithmic transformation improved the distribution of salivary alpha-amylase, although square root transformation was insufficient. The day-to-day reproducibility of these indices was assessed using intraclass correlation coefficients. Intraclass correlation coefficients of most heart rate variability and salivary indices were approximately 0.5 to 0.6. Intraclass correlation coefficients of raw salivary markers were approximately 0.6, which was similar to those of heart rate variability; however, log transformation of the salivary markers did not considerably improve their reproducibility. Correlations between sympathetic indicators of heart rate variability and salivary alpha-amylase were not observed. Conclusion Because the sample population examined in this study involved limited age and gender variations, the present results were independent of these factors and were indicative of pure interindividual variation.

2012-01-01

65

High salivary alpha-amylase levels in patients with schizophrenia: A pilot study  

Microsoft Academic Search

Previous studies have demonstrated the autonomic dysregulation in patients with schizophrenia using electrophysiological methods, such as electrodermal measures and heart rate analysis. Several theories have been proposed to explain the underlying mechanisms of schizophrenia and its autonomic function. Recently, the measurement of salivary alpha-amylase has been considered to be a useful tool for evaluating the sympathetic-adrenal-medullary (SAM) system. Psychosocial stress

Takuji Inagaki; Tsuyoshi Miyaoka; Shihoh Okazaki; Hideaki Yasuda; Tetsuya Kawamukai; Etsuko Utani; Rei Wake; Maiko Hayashida; Jun Horiguchi; Seiichi Tsuji

2010-01-01

66

Taking the starch out of oral biofilm formation: molecular basis and functional significance of salivary ?-amylase binding to oral streptococci.  

PubMed

?-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

Nikitkova, Anna E; Haase, Elaine M; Scannapieco, Frank A

2013-01-01

67

Taking the Starch out of Oral Biofilm Formation: Molecular Basis and Functional Significance of Salivary ?-Amylase Binding to Oral Streptococci  

PubMed Central

?-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.

Nikitkova, Anna E.; Haase, Elaine M.

2013-01-01

68

Salivary ?-amylase exhibits antiproliferative effects in primary cell cultures of rat mammary epithelial cells and human breast cancer cells  

PubMed Central

Background Breast cancer is one of the most diagnosed cancers in females, frequently with fatal outcome, so that new strategies for modulating cell proliferation in the mammary tissue are urgently needed. There is some, as yet inconclusive evidence that ?-amylase may constitute a novel candidate for affecting cellular growth. Methods The present investigation aimed to examine if salivary ?-amylase, an enzyme well known for the metabolism of starch and recently introduced as a stress marker, is able to exert antiproliferative effects on the growth of mammary gland epithelial cells. For this purpose, primary epithelial cultures of breast tissue from two different inbred rat strains, Fischer 344 (F344) and Lewis, as well as breast tumor cells of human origin were used. Treatment with human salivary ?-amylase was performed once daily for 2 days followed by cell counting (trypan blue assay) to determine alterations in cell numbers. Cell senescence after ?-amylase treatment was assessed by ?-galactosidase assay. Endogenous ?-amylase was detected in cells from F344 and Lewis by immunofluorescence. Results Salivary ?-amylase treatment in vitro significantly decreased the proliferation of primary cells from F344 and Lewis rats in a concentration-dependent manner. Noticeably, the sensitivity towards ?-amylase was significantly higher in Lewis cells with stronger impact on cell growth after 5 and 50 U/ml compared to F344 cells. An antiproliferative effect of ?-amylase was also determined in mammary tumor cells of human origin, but this effect varied depending on the donor, age, and type of the cells. Conclusions The results presented here indicate for the first time that salivary ?-amylase affects cell growth in rat mammary epithelial cells and in breast tumor cells of human origin. Thus, ?-amylase may be considered a novel, promising target for balancing cellular growth, which may provide an interesting tool for tumor prophylaxis and treatment.

2011-01-01

69

Interaction of Salivary alpha-Amylase and Amylase-Binding-Protein A (AbpA) of Streptococcus gordonii with Glucosyltransferase of S. gordonii and Streptococcus mutans  

Microsoft Academic Search

Background  Glucosyltransferases (Gtfs), enzymes that produce extracellular glucans from dietary sucrose, contribute to dental plaque\\u000a formation byStreptococcus gordoniiandStreptococcus mutans. The alpha-amylase-binding protein A (AbpA) ofS. gordonii, an early colonizing bacterium in dental plaque, interacts with salivary amylase and may influence dental plaque formation\\u000a by this organism. We examined the interaction of amylase and recombinant AbpA (rAbpA), together with Gtfs ofS. gordoniiandS.

Biswendu Chaudhuri; Jennifer Rojek; M Margaret Vickerman; Jason M Tanzer; Frank A Scannapieco

2007-01-01

70

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

PubMed Central

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

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

2013-01-01

71

Salivary Alpha-amylase and Cortisol in Toddlers: Differential Relations to Affective Behavior  

PubMed Central

This study applies a non-invasive and multi-system measurement approach (using salivary analytes) to examine associations between the psychobiology of the stress response and affective behavior in toddlers. Eighty-seven two-year-olds (48 females) participated in laboratory tasks designed to elicit emotions and behavior ranging from pleasure/approach to fear/withdrawal. Saliva samples were collected pre-task and immediately post-task, and assayed for markers of sympathetic nervous system (alpha-amylase or sAA) and hypothalamic-pituitary-adrenal axis (cortisol) activity. Individual differences in sAA were positively associated with approach behavior and positive affect; whereas, cortisol was positively associated with negative affect and withdrawal behavior. The findings suggest that individual differences in sAA may covary specifically with positive affect and approach behaviors or the predominant emotional state across a series of tasks. The results are discussed with respect to advancing biosocial models of the concomitants and correlates of young children’s affective behaviors.

Fortunato, Christine K.; Dribin, Amy E.; Granger, Douglas A.; Buss, Kristin A.

2008-01-01

72

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

PubMed Central

This research examined whether variations in salivary measures of the hypothalamic-pituitary-adrenal axis (cortisol) and autonomic nervous system (alpha amylase [sAA]) contribute to individual differences in the association between peer victimization and aggression. Children (N = 132; M age = 9.46 years, SD = .33) completed a measure of peer victimization, teachers rated children’s aggression, and children’s saliva was collected prior to, and following, participation in a laboratory-based peer-oriented social challenge task. Children rated their level of frustration at the end of the task. Results revealed that victimization interacted with cortisol and sAA measured in anticipation of the task to predict aggression; the victimization × cortisol contribution to aggression was partly mediated by children’s self-reported frustration level. Victimization also was associated with heightened frustration in girls with high task-related sAA reactivity. Task-related sAA reactivity was associated with heightened aggression, but only for girls. These findings suggest that associations between peer victimization and aggression are moderated by variation in the activity of the major components of the psychobiology of stress; results are discussed in relation to theoretical models of individual differences in biological sensitivity to context.

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

2011-01-01

73

Activated effect of lignin on ?-amylase.  

PubMed

This paper reports a new kind of activator of ?-amylase, lignin, which can greatly increase ?-amylase activity. The promoted ratio of lignin is even much higher than that of chloride ion, the traditional activator of ?-amylase. Further experimental results reveal that lignin may interact with ?-amylase to form a 1:1 complex with a binding constant of 4.47×10(5) M(-1). The binding is spontaneous and lignin/?-amylase complex formation is an exothermal reaction. Hydrogen bonding plays a key role and non-radiation energy transfers from ?-amylase to lignin in the binding process. Lignin, combining with ?-amylase, conforms to a first-order exponential decay function. The formation of the lignin/?-amylase complex results in the reduction of ?-helical content from 57.7% to 53.9%, the increase of the polarity around tryptophan residues, the decrease of the hydrophobicity, and the enlargement of protein granule volume. This work will give a deeper insight into lignin as a kind of dietary fibre, known as an important food functional factor. Furthermore, it also contributes to the exploration of an activator of ?-amylase, used in the food industry. PMID:23870952

Zhang, Juan; Cui, Jun-Hui; Yin, Tingting; Sun, Lizhou; Li, Genxi

2013-12-01

74

Sociodemographic risk, parenting, and effortful control: relations to salivary alpha-amylase and cortisol in early childhood.  

PubMed

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 levels. Findings support the open nature of stress responsive physiological systems to influence by features of the early caregiving environment and underscore the utility of including measures of these systems in prevention trials designed to influence child outcomes by modifying parenting behavior. PMID:22949301

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

2013-12-01

75

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

Microsoft Academic Search

This research examined whether variations in salivary measures of the hypothalamic-pituitary-adrenal axis (cortisol) and autonomic\\u000a nervous system (alpha amylase [sAA]) contribute to individual differences in the association between peer victimization and\\u000a aggression. Children (N?=?132; M age?=?9.46 years, SD?=?0.33) completed a measure of peer victimization, teachers rated children’s aggression, and children’s saliva was collected\\u000a prior to, and following, participation in a laboratory-based

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

2010-01-01

76

Efficient expression, purification and characterization of mouse salivary alpha-amylase secreted from methylotrophic yeast, Pichia pastoris.  

PubMed

We constructed a secretion vector of mouse salivary alpha-amylase, pPAM, using the AOX1 promoter-terminator and the secretion signal of 128 kDa pGKL killer protein, for an alternative yeast, Pichia pastoris. Taking advantage of multicopy insertion of the expression cassette and optimized growth conditions, we succeeded in highly efficient extracellular production (approximately 240 microg/ml) of mouse alpha-amylase in the 10 ml scale by conventional flask culture: this efficiency was about 90-fold higher than that of Saccharomyces cerevisiae. Growth temperature of cells was critical for efficient production of alpha-amylase. P. pastoris transformants secreted both core-glycosylated and non-glycosylated alpha-amylase molecules with a glycosylated:non-glycosylated ratio of about 20:80. Both glycosylated and non-glycosylated alpha-amylases were purified separately to apparent homogeneity. The signal sequence was correctly processed in both species, and the molecular masses of glycosylated and non-glycosylated alpha-amylase were determined to be 58 600 and 56 300, respectively, by mass spectrometry. We further studied the outer chain glycosylation of engineered mouse alpha-amylase secreted by P. pastoris. PMID:11329174

Kato, S; Ishibashi, M; Tatsuda, D; Tokunaga, H; Tokunaga, M

2001-05-01

77

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

PubMed

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

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

2014-05-01

78

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

PubMed Central

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

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

2009-01-01

79

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

Microsoft Academic Search

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

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

2010-01-01

80

Assessing Agreement Between Salivary Alpha Amylase Levels Collected by Passive Drool and Eluted Filter Paper in Adolescents With Cancer  

PubMed Central

Purpose/Objectives To assess the validity of filter paper (FP) against the gold standard of passive drool (PD) for collecting salivary alpha amylase as a surrogate biomarker of psychological stress in adolescents with cancer. Design Part of a longitudinal, descriptive study of symptoms in adolescents with cancer during chemotherapy. Setting A pediatric hematology/oncology treatment center. Sample 33 saliva sample pairs from nine adolescents with cancer, aged 13–18 years. Methods Salivary alpha amylase was collected by PD and FP at four time points during a cycle of chemotherapy: days 1 (time 1) and 2 (time 2) of chemotherapy, day 7–10 (time 3), and day 1 of the next cycle (time 4). A random effects regression was used to assess the correlation between PD and FP values, and a Bland Altman analysis was conducted to assess agreement between the values. Main Research Variables Salivary alpha amylase. Findings The estimated correlation between PD and FP values was r = 0.91, p < 0.001. Regression results were also used to rescale FP values to the levels of the PD values because the FP values were on a different scale than the PD values. The Bland Altman analysis revealed that the agreement between the rescaled FP values and PD values was not satisfactory. Conclusions Eluted FP may not be a valid method for collecting salivary alpha amylase in adolescents with cancer. Implications for Nursing Psychological stress in adolescents with cancer may be linked to negative outcomes, such as greater symptom severity and post-traumatic stress disorder. Nurses need valid, efficient, biobehavioral measures to assess psychological stress in the clinical setting.

Ameringer, Suzanne; Munro, Cindy; Elswick, R.K.

2014-01-01

81

Effect of mobile phone use on salivary concentrations of protein, amylase, lipase, immunoglobulin A, lysozyme, lactoferrin, peroxidase and C-reactive protein of the parotid gland.  

PubMed

Background: The possibility of side effects associated with the electromagnetic waves emitted from mobile phones is a controversial issue. The present study aimed to evaluate the effect of mobile phone use on parotid gland salivary concentrations of protein, amylase, lipase, immunoglobulin A, lysozyme, lactoferrin, peroxidase and C-reactive protein. Methods: Stimulated salivary samples were collected simultaneously from both parotid glands of 86 healthy volunteers. Salivary flow rate and salivary concentrations of proteins, amylase, lipase, lysozyme, lactoferrin, peroxidase, C-reactive protein and immunoglobulin A, were measured. Data were analysed using t-tests and one-way analyses of variance. Results: Salivary flow rate and parotid gland salivary concentrations of protein were significantly higher on the right side compared to the left in those that predominantly held mobile phones on the right side. In addition, there was a decrease in concentrations of amylase, lipase, lysozyme, lactoferrin and peroxidase. Conclusion: The side of dominant mobile phone use was associated with differences in salivary flow rate and parotid gland salivary concentrations, in right-dominant users. Although mobile phone use influenced salivary composition, the relationship was not significant. PMID:24739140

Hashemipour, M S; Yarbakht, M; Gholamhosseinian, A; Famori, H

2014-05-01

82

Kinetic measurement of total amylase and isoamylase activities with a centrifugal analyzer.  

PubMed

We evaluated a new kinetic assay for alpha-amylase (Phadebas IsoAmylase Test), using modified starch as the substrate and a CentrifiChem 400 centrifugal analyzer system. We determined isoamylase activities by using a selective inhibitor. Results were compared with those obtained with the chromogenic Phadebas dyed-starch technique. The method for total amylase appeared to be rapid and precise (CV = 4%) and results correlated well with the chromogenic method. Samples with activities up to 3000 arb. units/L can be analyzed without dilution. Glucose and pyruvate interfere with the assay, but hemoglobin and bilirubin do not. Pancreatic (P) and salivary (S) isoamylase activity can be determined with acceptable precision (CVP = 8%, CVS = 10%) by an automated routine procedure with commercially available reagents. PMID:6189640

de Rijke, D; Kreutzer, H J

1983-06-01

83

Inhibition of ?-amylase activity by condensed tannins  

Microsoft Academic Search

The ability of grape seed procyanidins to inhibit ?-amylase activity was studied using a colorimetric method. This ability was found to be related with the average degree of polymerisation of the tested procyanidins. These interactions were further evaluated by fluorescence quenching, dynamic light scattering (DLS) and nephelometry, in order to understand the mechanisms by which they occur. A relationship between

Rui Gonçalves; Nuno Mateus; Victor de Freitas

2011-01-01

84

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

PubMed Central

We examined daytime salivary cortisol and salivary alpha-amylase (sAA) secretion levels and variability in preschool-aged children with autism (AUT) and typically developing children (TYP). Fifty-two subjects (26 AUT and 26 TYP) were enrolled. Salivary samples were obtained at waking, midday, and bedtime on two consecutive days at three phases (baseline, 3 months later, 6 months later). There were modest increases in waking cortisol and sAA levels in AUT relative to TYP, but the increases were not statistically significant. Important differences were observed in cortisol and sAA variability between AUT and TYP. There was also a graded response among AUT by functional status—cortisol and sAA secretion levels were higher when IQ was lower.

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

2013-01-01

85

Possible mechanisms of normal amylase activity in hyperlipemic pancreatitis.  

PubMed

Lipemic serum from three patients with acute pancreatitis and type IV hyperlipemia was fractionated into very-low-density lipoproteins and clear serum. Amylase activity (determined by the Phadebas method) in the component fractions did not exceed that in the original lipemic serum. Addition of these fractions or VLDL and chylomicrons from asymptomatic patients with hyperlipemia to nonlipemic serum from patients with "routine acute pancreatitis" did not inhibit amylase activity or alter the electrophoretic mobility of amylase isoenzymes. Therefore the normal amylase activity often observed in hyperlipemic pancreatitis does not result from an inhibition of amylase activity by serum lipoproteins. PMID:206333

Mishkin, S; Bates, J; O'Hashi, J; Schneider, P; Sniderman, A D; Wolf, R O

1976-11-20

86

DEVELOPMENTAL VALIDATION OF A POINT-OF-CARE, SALIVARY ?-AMYLASE BIOSENSOR  

PubMed Central

The translation of salivary alpha-amylase (sAA) to the ambulatory assessment of stress hinges on the development of technologies capable of speedy and accurate reporting of sAA levels. Here, we describe the developmental validation and usability testing of a point-of-care, colorimetric, sAA biosensor. A disposable test strip allows for streamlined sample collection and a corresponding hand-held reader with integrated analytic capabilities permits rapid analysis and reporting of sAA levels. Bioanalytical validation utilizing saliva samples from 20 normal subjects indicates that, within the biosensor’s linear range (10–230 U/ml), its accuracy (R2 = 0.989), precision (CV < 9%), and measurement repeatability (range ?3.1% to + 3.1%) approach more elaborate laboratory-based, clinical analyzers. The truncated sampling-reporting cycle (< 1 minute) and the excellent performance characteristics of the biosensor has the potential to take sAA analysis out of the realm of dedicated, centralized laboratories and facilitate future sAA biomarker qualification studies.

Shetty, Vivek; Zigler, Corwin; Robles, Theodore F.; Elashoff, David; Yamaguchi, Masaki

2010-01-01

87

Developmental differences in infant salivary alpha-amylase and cortisol responses to stress  

PubMed Central

Summary This study examined developmental differences in infants’ salivary alpha-amylase (sAA) and cortisol levels and responses to the well-baby exam/inoculation stress protocol at 2, 6, 12, and 24 months of age. Mother-infant pairs (N = 85; 45 girls) were assessed during well-baby visits and saliva was sampled before the well baby exam/inoculation procedure (pretest) and at 5, 10, and 20 minutes post inoculation stress. Older infants (24 months) had higher levels of sAA than younger infants (2, 6 and 12 months). Stress-related sAA increases were evident at 6 and 12 months, but not at 2 or 24 months of age. Stress-related cortisol increases were present at 2 and 6 months, but not at older ages. Mothers had higher sAA levels than their infants, but did not show sAA or cortisol increases to their infants’ inoculation. Pre-test, maternal and infant sAA levels were positively correlated (rs.47 to.65) at 6, 12, and 24 months of age, but not at 2 months. These findings suggest that the association between the sympathetic branch of the autonomic nervous system and the secretion of sAA develops between 2 and 6 months of age, when levels of sAA are responsive to exposure to a painful stressor.

Davis, Elysia Poggi; Granger, Douglas A.

2009-01-01

88

Variation in salivary and pancreatic alpha-amylase genes in italian horse breeds.  

PubMed

The dietary demand of the modern horse relies on high-cereal feeding and limited forage compared with natural grazing conditions, predisposing the horse to several important diseases. Salivary and pancreatic alpha-amylases (coded by AMY1 and AMY2 genes, respectively) play a crucial role in carbohydrate digestion in nonruminants, but little is known about these 2 genes in the horse. Aim of this work has been to distinguish genomic sequences of horse AMY1 and AMY2 genes and to analyze any polymorphisms in breeds historically characterized by marked differences in nutritional management. A single nucleotide polymorphism detection was performed and 7 novel single nucleotide polymorphisms were found. Three single nucleotide polymorphisms are in exons and were genotyped in 112 horses belonging to 6 breeds. One single nucleotide polymorphism in AMY1 gene distinguished Haflinger and the Italian native Murgese from the other breeds, whereas both the single nucleotide polymorphisms in AMY2 gene showed different allelic frequencies in Friesian compared with the other breeds. These differences are confirmed by quite high fixation index (Fst) values for these 2 nonsynonymous single nucleotide polymorphisms. These preliminary results highlight marked divergences in allele frequencies of AMY1 and AMY2 genes, involved in starch digestion, between horse breeds characterized by different histories of selection, thus providing first indications of possible relations between genetics and nutritional management. PMID:24558100

Coizet, Beatrice; Nicoloso, Letizia; Marletta, Donata; Tamiozzo-Calligarich, Alessandra; Pagnacco, Giulio; Crepaldi, Paola

2014-01-01

89

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

PubMed

Human salivary alpha-amylase (HSA) is a major secretory protein component of saliva and has important biological functions, including the initial digestion of starch. HSA acts as a monomer and mediates the hydrolysis of alpha-1,4-glucosidic linkages in oligosaccharides. To date, all published crystal structures of HSA have been crystallized as monomers in space group P2(1)2(1)2(1). Here, the serendipitous purification, crystallization and ultimate structure determination of a HSA non-crystallographic symmetry (NCS) dimer, while attempting to purify human carbonic anhydrase VI (HCA VI) from saliva using an affinity resin for alpha-class carbonic anhydrases, is presented. On further investigation, it was shown that HSA could only be copurified using the affinity resin in the presence of HCA VI which is glycosylated and not the non-glycosylated HCA II. The identification of the HSA crystals was carried out by peptide mapping and mass spectrometry. HSA was shown to have crystallized as an NCS dimer in space group C2, with unit-cell parameters a = 150.9, b = 72.3, c = 91.3 A, beta = 102.8 degrees. The NCS dimer crystal structure is reported to 3.0 A resolution, with a refined Rcryst of 0.228. The structure is compared with the previously reported P2(1)2(1)2(1) monomer structures and the crystal packing and dimer interface are discussed. PMID:16511271

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

2006-02-01

90

The sensitivity and specificity of the RSID-saliva kit for the detection of human salivary amylase in the Forensic Science Laboratory, Dublin, Ireland.  

PubMed

We demonstrate here that the RSID-saliva test can be used as a test for human salivary alpha-amylase on samples routinely examined in forensic casework. We show that the RSID-saliva test detects salivary alpha-amylase at lower concentrations than the Phadebas Quantitative test, that the RSID-saliva test does not cross-react with forensically important human fluids and that the RSID-saliva test can be successfully integrated into the whole swab semen extraction method. PMID:19931992

Casey, David G; Price, Judy

2010-01-30

91

Simultaneous expression of salivary and pancreatic amylase genes in cultured mouse hepatoma cells.  

PubMed Central

The tissue-specific expression of two types of mouse amylase genes does not overlap in vivo; the Amy-1 locus is transcribed in the parotid gland and the liver, while expression of Amy-2 is limited to the pancreas. We identified a mouse hepatoma cell line, Hepa 1-6, in which both amylase genes can be simultaneously expressed. Amy-1 is constitutively active in these cells and is inducible by dexamethasone at the level of mRNA. We demonstrated that the liver-specific promoter of Amy-1 is utilized by the dexamethasone-treated hepatoma cells, and that glucocorticoid consensus sequences are present upstream of this promoter. Amy-2 is not detectable constitutively, but can be activated if the cells are cultured in serum-free medium containing dexamethasone. Expression of Amy-2 in a nonpancreatic cell type has not previously been observed. We speculate that induction of Amy-1 and activation of Amy-2 may involve different regulatory mechanisms. Hepa 1-6 cells provide an experimental system for molecular analysis of these events. Images

Darlington, G J; Tsai, C C; Samuelson, L C; Gumucio, D L; Meisler, M H

1986-01-01

92

The salivary alpha amylase over cortisol ratio as a marker to assess dysregulations of the stress systems.  

PubMed

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

Ali, Nida; Pruessner, Jens C

2012-04-12

93

Tissue-specific expression of mouse-alpha-amylase genes: nucleotide sequence of isoenzyme mRNAs from pancreas and salivary gland.  

PubMed

We have determined the nucleotide sequence of two different mouse alpha-amylase mRNAs, one found in the pancreas and the other in the salivary gland. The 1577 and 1659 nucleotide mRNAs from pancreas and salivary gland, respectively are the major alpha-amylase species which accumulate in each tissue. Differences in mRNA length are primarily in the 5' noncoding regions. Comparable portions of the mRNAs are 89% homologous. The mRNA sequences predict alpha-amylase precursor proteins of 508 and 511 amino acid residues, accounting for nearly the entire coding capacity of the mRNAs; differences in protein length occur as a result of a nine nucleotide segment present within the translated portion of salivary gland, but not pancreas, mRNA. The lengths and amino acid compositions of the predicted proteins concur with those determined empirically by others. These proteins differ 12% in amino acid sequence, explaining previously observed differences in net charge and antigenic properties. Finally, translation of the salivary gland alpha-amylase mRNA is not initiated at the AUG codon nearest the 5' terminus since that codon is almost immediately followed by the termination triplet UAA. This observation may have implications for the mechanism of translation initiation in eucaroytes. PMID:6157477

Hagenbüchle, O; Bovey, R; Young, R A

1980-08-01

94

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

PubMed

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

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

2013-01-01

95

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

PubMed

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

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

2014-05-01

96

The Relationship Between Cortisol, Salivary Alpha-Amylase, and Cognitive Bias in Young Women  

Microsoft Academic Search

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

Donna A. Kreher; Sally I. Powers; Douglas A. Granger

2012-01-01

97

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

SciTech Connect

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

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

1991-02-01

98

Automated measurement of amylase isoenzymes with 4-nitrophenyl-maltoheptaoside as substrate and use of a selective amylase inhibitor.  

PubMed

We automated a kinetic procedure for determining amylase isoenzymes in serum and urine samples. We used 4-nitro-phenylmaltoheptaoside as substrate and a selective amylase inhibitor with the Abbott-VP bichromatic system. By use of the maximum differences between pancreatic (P) and salivary (S) amylase activities remaining after inhibition by the selective inhibitor and by use of the linear range, a one-point standard method for calibration is proposed for determining amylase activities between about 50 and 1500 U/L when the P/S ratio exceeds 0.2. Results correlated well with those by electrophoresis and the Phadebas method (r = 0.99 for both pancreatic and salivary amylase). Reproducibilities (CVs) were 1.5% to 5.5% for pancreatic amylase and 1.4% to 3.3% for salivary amylase in serum, 0.8% to 2.0% for pancreatic amylase and 0.8% to 2.3% for salivary amylase in urine. PMID:6203667

Okabe, H; Uji, Y; Netsu, K; Noma, A

1984-07-01

99

A new potentiometric sensor for the determination of ?-amylase activity.  

PubMed

A platinum redox sensor for the direct potentiometric determination of ?-amylase concentration has been described. The sensor measured the amount of triiodide released from a starch-triiodide complex, which was correlated with the ?-amylase activity after biocatalytic starch degradation. The composition and stability of the potassium triiodide solution was optimized. The starch-triiodide complex was characterized potentiometrically at variable starch and triiodide concentrations. The response mechanism of the platinum redox sensor towards ?-amylase was proposed and the appropriate theoretical model was elaborated. The results obtained using the redox sensor exhibited satisfactory accuracy and precision and good agreement with a standard spectrophotometric method and high-sensitive fully automated descret analyser method. The sensor was tested on pure ?-amylase (EC 3.2.1.1, Fluka, Switzerland), industrial granulated ?-amylase Duramyl 120 T and an industrial cogranulate of protease and ?-amylase Everlase/Duramyl 8.0 T/60 T. The detection limit was found to be 1.944 mU for ?-amylase in the range of 0-0.54 U (0-15 ?g), 0.030 mKNU for Duramyl 120 T in the range of 0-9.6 mKNU (0-80 ?g) and 0.032 mKNU for Everlase/Duramyl 8.0 T/60 T in the range of 0-9.24 mKNU (0-140 ?g). PMID:21238759

Sakac, Nikola; Sak-Bosnar, Milan; Horvat, Marija; Maduni?-Caci?, Dubravka; Szechenyi, Aleksandar; Kovacs, Barna

2011-02-15

100

Amylase activity of Torulopsis ingeniosa Di Menna.  

PubMed

Torulopsis ingeniosa DI MENNA was found to possess an alpha-amylase strongly attached to the cell wall, its pH optimum being at 5.5, optimum temperature at 50 degrees C, highly sensitive to thermal inactivation. The enzyme was found to be induced by starch but the synthesis is not subject to a glucose effect. PMID:33881

Moulin, G; Galzy, P

1978-01-01

101

A novel reusable PAni-PVA-Amylase film: activity and analysis.  

PubMed

Alpha-amylase was immobilized onto polyaniline-polyvinyl alcohol (PAni-PVA) film by cross linking with glutaraldehyde. The activity yield and immobilization efficiency was calculated as 13.58% and 30.60% respectively. The PAni-PVA-Amylase films were characterized by Fourier Transform Infrared Spectroscopy (FTIR) and amylase activity assay. The immobilized ?-amylase had its optimum activity at pH 7.0 and 25°C. The immobilization of ?-amylase onto the PAni-PVA films protected the enzyme from Fe(2+) inhibition. The PAni-PVA-Amylase films could be used effectively for 30 cycles with 60% retention of the activity. PMID:23434690

Singh, Susmita; Saikia, Jyoti Prasad; Buragohain, Alak Kumar

2013-06-01

102

A novel electrochemical method to determine ?-amylase activity.  

PubMed

In this paper, we report a novel electrochemical method that can be developed as a biosensor for simple and direct determination of ?-amylase activity. The method is based on the hydrolysis of maltopentaose, the substrate of the enzyme, which is immobilized on the surface of a gold electrode, and the induced charge changes of the substrate-modified electrode. Specifically, the substrate maltopentaose is immobilized onto a gold electrode surface via a simple and direct immobilization technique that involves a one-step and site-specific attachment of unmodified maltopentaose to the hydrazide-derivatized surface. So, by analyzing the electrochemical signal obtained from the electro-active molecule [Ru(NH3)5Cl](2+) during the hydrolysis of maltopentaose, the determination of ?-amylase activity is achieved. Under optimized conditions, ?-amylase activity can be assayed with a detection limit of 0.022 U mL(-1). The biosensor exhibits a rapid response, good stability and anti-interference ability. Furthermore, the biosensor has also been successfully applied to detect ?-amylase in human serum, which shows acceptable accuracy compared to the currently used clinical method. The proposed method in this work may also have potential application of ?-amylase determination in real blood samples, diagnostics and food production in the future. PMID:24855635

Zhang, Juan; Cui, Junhui; Liu, Ying; Chen, Yangyang; Li, Genxi

2014-07-01

103

The feasibility of ambulatory biosensor measurement of salivary alpha amylase: Relationships with self-reported and naturalistic psychological stress  

PubMed Central

Summary Recent developments in biosensor technology allow point-of-use reporting of salivary alpha amylase (sAA) levels while approaching the precision and accuracy of conventional laboratory-based testing. We deployed a portable prototype sAA biosensor in 54 healthy, male dental students during a low stress baseline and during final exams. At baseline, participants completed the Brief Symptom Inventory (BSI). At baseline and the exam week, participants provided saliva samples at 10 AM, 1 PM, and 5 PM, and rated concurrent subjective distress. Although subjective distress was higher during exams compared to baseline, sAA levels did not differ between baseline and exams. Higher sAA levels were related to higher concurrent subjective distress, and higher depressive and social isolation symptoms on the BSI were related to lower sAA during exams. Results from this study, in combination with previous validation data, suggest that the sAA biosensor is a promising tool for point-of-use measures of exposure to stress.

Robles, Theodore F.; Shetty, Vivek; Zigler, Corwin M.; Glover, Dorie A.; Elashoff, David; Murphy, Debra; Yamaguchi, Masaki

2010-01-01

104

Vasodilatory activity in horsefly and deerfly salivary glands  

Microsoft Academic Search

Salivary gland extract (SGE) of four horsefly species (Hybomitra bimaculata Macquart, Hybomitra ciureai Seguy, 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

P. R AJ; SK A ´; O. P E C H ANOV; P. T A K AC Ay; M. K A Z I M IROVA ´; L. R OLLER; P. A. N UTTALL

2003-01-01

105

Ixodes scapularis: salivary kininase activity is a metallo dipeptidyl carboxypeptidase.  

PubMed

Saliva and salivary gland homogenates of Ixodes scapularis contain a dipeptidyl carboxypeptidase activity that accounts for the previously described salivary kininase activity of this tick. Reversed phase HPLC and laser desorption mass spectrography of the reaction products identified bradykinin fragment 1-7 and 1-5 as being produced subsequent to incubation of purified salivary kininase with bradykinin. The activity was inhibited by captopril and EDTA and was activated by cobalt and manganese, a behavior similar to that displayed by angiotensin-converting enzymes of vertebrate and invertebrate origins. PMID:9635445

Ribeiro, J M; Mather, T N

1998-06-01

106

Allele-Dependent Barley Grain ?-Amylase Activity1  

PubMed Central

The wild ancestor of cultivated barley, Hordeum vulgare subsp. spontaneum (K. Koch) A. & Gr. (H. spontaneum), is a source of wide genetic diversity, including traits that are important for malting quality. A high ?-amylase trait was previously identified in H. spontaneum strains from Israel, and transferred into the backcross progeny of a cross with the domesticated barley cv Adorra. We have used Southern-blot analysis and ?-amy1 gene characterization to demonstrate that the high ?-amylase trait in the backcross line is co-inherited with the ?-amy1 gene from the H. spontaneum parent. We have analyzed the ?-amy1 gene organization in various domesticated and wild-type barley strains and identified three distinct ?-amy1 alleles. Two of these ?-amy1 alleles were present in modern barley, one of which was specifically found in good malting barley cultivars. The third allele, linked with high grain ?-amylase activity, was found only in a H. spontaneum strain from the Judean foothills in Israel. The sequences of three isolated ?-amy1 alleles are compared. The involvement of specific intron III sequences, in particular a 126-bp palindromic insertion, in the allele-dependent expression of ?-amylase activity in barley grain is proposed.

Erkkila, Maria J.; Leah, Robert; Ahokas, Hannu; Cameron-Mills, Verena

1998-01-01

107

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

PubMed

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

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

2013-01-01

108

Salivary proteins promote proteolytic activity in Streptococcus mitis biovar 2 and Streptococcus mutans.  

PubMed

A major function of the salivary pellicle on oral surfaces is to promote colonization of the commensal microbiota by providing binding sites for adherence. Streptococcus mitis is an early colonizer of the oral cavity whereas Streptococcus mutans represents a later colonizer. To survive and grow, oral bacteria produce enzymes, proteases and glycosidases, which allow them to exploit salivary proteins as a nutrient source. In this study, adherence and proteolytic activity of S. mitis biovar 2 and S. mutans were investigated in a flow-cell model in the presence of different populations of surface-associated salivary proteins. Streptococcus mitis biovar 2 adhered well to surfaces coated with both a MUC5B-enriched fraction and a pool of low-density proteins containing MUC7, amylase, cystatin, gp340, immunoglobulin A, lactoferrin, lysozyme and statherin, whereas adherence of S. mutans to these proteins was poor. In environments of MUC5B or the low-density proteins, both S. mitis biovar 2 and S. mutans showed high levels of proteolytic activity. For S. mitis in the MUC5B environment, most of this activity may be attributable to contact with the molecules in the fluid phase although activity was also enhanced by adherence to surface-associated MUC5B. These data suggest that although they differ in their capacity to adhere to surface-associated salivary proteins, in the natural environment exploitation of saliva as a nutrient source can contribute to survival and colonization of the oral cavity by both S. mitis biovar 2 and S. mutans. PMID:22958385

Kindblom, C; Davies, J R; Herzberg, M C; Svensäter, G; Wickström, C

2012-10-01

109

Action pattern of serum amylase using p-nitrophenyl-maltoheptaoside as substrate.  

PubMed

Selected serum samples with different isoamylase patterns, as revealed by agarose gel electrophoresis, were analysed by the Phadebas amylase test, and by using p-nitrophenyl-maltoheptaoside as the substrate. The products with the latter substrate were examined by high performance liquid chromatography (HPLC). For some samples, different catalytic activities were registered by the two amylase methods. In these cases, electrophoresis showed a double pancreatic band associated with increased pancreatic amylase catalytic activity, or an abnormal, broad salivary band associated with increased salivary enzyme catalytic activity. The stoichiometric factors for most of these samples, calculated from the amount of p-nitrophenol detected after HPLC, differed from the mean values. PMID:6207265

Masson, P; Hultberg, B

1984-06-01

110

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

PubMed

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

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

2014-04-01

111

Salivary Ceruloplasmin Ferroxidase & Oxidase Activities in Celiac Patients  

PubMed Central

The aim of the current study was to evaluate salivary ferroxidase ceruloplasmin activities in celiac patients with different histopathological severity. This study included 75 celiac patients with different mean age (18.68 ± 11.13) year, who had positive screen for celiac antibodies, and who had gastrointestinal symptoms. In order to simplify the comparison with the healthy control group, celiac patients were divided into two groups according to their histopathological severity: severe (marsh IIIa, b, c) & less severe (marsh 0, I). All these patients have been evaluating for salivary ceruloplasmin (Cp) concentration and Cp ferroxidase activities. To confirm the presence of the enzymatic activity of this protein, polyacrylamide gel electrophoresis was carried out and then stained for Cp ferroxidase, as well as for Cp oxidase activity. Furthermore, the concentrations of salivary total protein, albumin, and globulin were measured in the studied groups. A significant increase (p<0.05) in salivary concentration of ceruloplasmin was found in all above mentioned patients groups in comparison to that of the control group, except for total villous atrophy (marsh IIIc) patients subgroup. Salivary Cp ferroxidase activity revealed statistically significant decrease among the patient groups as well as between them and the control group. The result of salivary total protein and globulin showed presence a significant increase (p<0.05) in comparison to that of the control group. Meanwhile albumin levels was found to increase non-significantly (p=0.186).

Hasan, Hathama R.; Ghadhban, Jasim M.; Abudal Kadhum, Zahraa I.

2012-01-01

112

Catalytic concentrations of amylase isoenzymes: an assay with wheat-germ inhibitor and 4-nitrophenylmaltopentaoside plus 4-nitrophenylmaltohexaoside as substrate.  

PubMed

We coupled a kinetic procedure to a selective inhibiting method for determining amylase isoenzymes in biological samples, using 4-nitrophenylmaltopentaoside plus 4-nitrophenylmaltohexaoside as substrate and a wheat-germ selective inhibitor with the Gilford S-III spectrophotometer. On plotting remaining amylase activities/total amylase activities (R/T) vs pancreatic amylase activities/salivary amylase activities (P/S) ratios, we found the curve to be linear for P/S ratios from 0.2 to 5. The inhibition rate of amylase inhibitor was constant in solutions having total amylase activities between 20 and (at least) 900 U/L. CVs were 3.1 to 7.1% for pancreatic amylase and 2.0 to 12.9% for salivary amylase in serum. Correlation with the Phadebas method was excellent (r = 0.99) for both pancreatic and salivary amylase. We also automated this procedure in an Hitachi 705 analyzer and correlated the results (r = 0.99) with those by our manual method. PMID:2426010

Jiménez, A; Arenas, J; Santos, I; Martínez, A

1986-08-01

113

Probing the role of aromatic residues at the secondary saccharide binding sites of human salivary ?-amylase in substrate hydrolysis and bacterial binding  

PubMed Central

SUMMARY Human salivary ?-amylase (HSAmy) has three distinct functions relevant to oral health: 1) hydrolysis of starch; 2) binding to hydroxyapatite; and 3) binding to bacteria (e.g. viridans streptococci). Although the active site of HSAmy for starch hydrolysis is well characterized, the regions responsible for the bacterial binding are yet to be defined. Since HSAmy possesses several secondary saccharide-binding sites in which aromatic residues are prominently located, we hypothesized that one or more of the secondary saccharide binding sites harboring the aromatic residues may play an important role in bacterial binding. To test this hypothesis, the aromatic residues at five secondary binding sites were mutated to alanine to generate six mutants representing either single (W203A, Y276A and W284A), double (Y276A/W284A and W316A/W388A) or multiple (HSAmy-ar; W134A/W203A/Y276A/W284A/W316A/W388A) mutations. The crystal structure of HSAmy-ar was determined at a resolution of 1.5 Ĺ as an acarbose complex and compared with the existing wild type acarbose complex. The wild type and the mutant enzymes were characterized for their abilities to exhibit enzyme activity, starch binding, hydroxyapatite and bacterial binding activities. Our results clearly showed that 1) mutation of aromatic residues does not alter the overall conformation of the molecule; 2) the single or double mutants showed either moderate or minimal changes in both starch and bacterial binding activities activity whereas the HSAmy-ar showed significant reduction in these activities; 3) the starch hydrolytic activity was reduced 10-fold in HSAmy-ar; 4) oligosaccharide hydrolytic activity was reduced in all the mutants but the action pattern was similar to that of the wild type enzyme; and 5) the hydroxyaptite binding was unaffected in HSAmy-ar. These results clearly show that the aromatic residues at the secondary saccharide binding sites in HSAmy play a critical role in bacterial binding and starch hydrolytic functions of HSAmy.

Ragunath, Chandran; Manuel, Suba G.A.; Venkataraman, Venkat; Sait, Hameetha B.R.; Kasinathan, Chinnasamy; Ramasubbu, Narayanan

2008-01-01

114

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

ERIC Educational Resources Information Center

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

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

2012-01-01

115

The ram1 mutant of Arabidopsis exhibits severely decreased beta-amylase activity.  

PubMed

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

Laby, R J; Kim, D; Gibson, S I

2001-12-01

116

Evaluation of amylase testing as a tool for saliva screening of crime scene trace swabs.  

PubMed

Amylase testing has been used as a presumptive test for crime scene saliva for over three decades, mainly to locate saliva stains on surfaces. We have developed a saliva screening application for crime scene trace swabs, utilising an amylase sensitive paper (Phadebas(®) Forensic Press test). Positive results were obtained for all tested dried saliva stains (0.5-32 ?L) with high or intermediate amylase activity (840 and 290 kU/L). Results were typically obtained within 5 min, and all samples that produced DNA profiles were positive. However, salivary amylase activities, as well as DNA concentrations, vary significantly between individuals. We show that there is no correlation between amylase activity and amount of DNA in fresh saliva. Even so, a positive amylase result indicates presence of saliva, and thereby presence of DNA. Amylase testing may be useful for screening in investigations where the number of DNA analyses is limited due to cost, e.g., in volume crime. PMID:20457099

Hedman, Johannes; Dalin, Erik; Rasmusson, Birgitta; Ansell, Ricky

2011-06-01

117

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

SciTech Connect

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

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

1990-06-01

118

Maltotetraose as a substrate for enzyme-coupled assay of amylase activity in serum and urine.  

PubMed

The use of maltotetraose ss a new substrate for the enzyme-coupled determination of amylase activity in biological fluids was developed by Beckman Microbics. We evaluated a manual and a centrifugal analyzer version of the method in comparison with two commonly used manual starch-dye amylase techniques: Roche Amylochrome and Pharmacia Phadebas. Both maltotetraose amylase procedures proved to be rapid and precise, and results correlated satisfactorily with the starch-dye methods for serum and urine samples. PMID:95558

Whitlow, K J; Gochman, N; Forrester, R L; Wataji, L J

1979-03-01

119

Activity and cellular localization of amylases of rabbit cecal bacteria  

Microsoft Academic Search

Five 11-week-old rabbits, fed a commercial granulated feed, were slaughtered and cecal starch-degrading bacteria enumerated;\\u000a total concentration of cultivable bacteria utilizing starch averaged 5.5 × 1010 CFU\\/g. The activity and cellular localization of amylases was determined in 9 bacteria identified asActinomyces israeli (strains AA2 and AD4),Bacteroides spp. (strain AA3),Dichelobacter nodosus (strain AA4),Mitsuokella multiacidus (strain AA6),Eubacterium spp. (strains AA7 and AB2),Clostridium

K. Sirotek; M. Marounek; O. Suchorská

2006-01-01

120

Serum amylase and isoamylase assay on the Hitachi 705 automatic clinical chemical analyzer.  

PubMed

The automated continuous alpha-amylase assay using p-nitrophenyl-alpha-D-maltoheptaoside (Boehringer Mannheim) as substrate on the Hitachi 705 clinical chemical analyzer, was compared with the Phadebas Kinetic Amylase Assay (Pharmacia Diagnostics) on the Hitachi 705. The two methods showed good correlation. The precision varied from 1.0 to 2.5% (CV) within-day and from 1.1 to 5.6% (CV) day-to-day. The substrate, p-nitrophenyl-alpha-D-maltoheptaoside, was also applied to an automated isoamylase assay. The amylase inhibitor from wheat was used to determine the ratio of pancreatic and salivary amylase activities of serum. About 80% of salivary type amylase was inhibited up to an activity level of 1000 U/l, while inhibition of pancreatic type amylase activity was only 10-15%. Ratios of pancreatic to salivary amylase from 0.1 to 10 can be evaluated in serum with wheat inhibitor. The precision of the isoamylase determination by the Boehringer amylase method was acceptable. PMID:6199455

Parviainen, M T; Koivula, T; Jokela, H

1984-01-01

121

Vasodilatory activity in horsefly and deerfly salivary glands.  

PubMed

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

Rajská, P; Pechánová, O; Takác, P; Kazimírová, M; Roller, L; Vidlicka, L; Ciampor, F; Labuda, M; Nuttall, P A

2003-12-01

122

Potent ?-amylase inhibitory activity of Indian Ayurvedic medicinal plants  

PubMed Central

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

2011-01-01

123

Antihemostatic activity in salivary glands of the human body louse, Pediculus humanus humanus (Anoplura: Pediculidae)  

Microsoft Academic Search

A thrombin inhibitor, factor Xa inhibitor and apyrase activity were demonstrated in the salivary glands of Pediculus humanus humanus. Less than 0.15 ?g of salivary gland extracts (SGE), which is equivalent to approximately 0.15 lice salivary glands, caused 50% inhibition of thrombin activity, while 1 ?l of SGE was sufficient to double thrombin time using rat plasma. A single peak

K. Y. Mumcuoglu; R. Galun; Y. Kaminchik; A. Panet; A. Levanon

1996-01-01

124

Hydrogen sulfide stimulates ?-amylase activity during early stages of wheat grain germination  

PubMed Central

We recently reported that H2S could significantly promote the germination of wheat grains subjected to aluminum (Al3+) stress.1 In these experiments seeds were pretreated with the H2S donor NaHS for 12 h prior to Al3+ stress. During this pre-incubation period we observed that H2S increased the activity of grain amylase in the absence of Al3+. Using embryoless half grains of wheat we now show that H2S preferentially affects the activity of endosperm ?-amylase and that ?-amylase synthesis and activity is unaffected by this treatment.

Dou, Wei; Jiang, Cheng-Xi; Wei, Zhao-Jun; Liu, Jian

2010-01-01

125

An analytical method for measuring ?-amylase activity in starch-containing foods.  

PubMed

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

Koyama, Kazuo; Hirao, Takashi; Toriba, Akira; Hayakawa, Kazuichi

2013-05-01

126

Determining the relationship of acute stress, anxiety, and salivary alpha-amylase level with performance of student nurse anesthetists during human-based anesthesia simulator training.  

PubMed

Managing stress for student nurse anesthetists represents a multifaceted educational concern for anesthesia educators. Our purpose was to determine the relationship between physiologic measures of stress and performance of student nurse anesthetists during anesthesia simulator training. Following institutional review board approval, 78 students were enrolled from a nurse anesthesia program. A prospective descriptive design was used to compare baseline, acute, and recovery measurements of stress with performance scores of students during an induction and intubation sequence in a patient simulator. Performance scores were stratified into low-, moderate-, and high-performing groups based on scores received from trained observers. A statistically significant difference in physiologic measures of stress was detected between baseline and acute levels of salivary a-amylase (P = .017), heart rate (P = .003), and anxiety levels (P = .001). No significant differences were found when measures of stress were compared with performance of low, moderate, or high performers. This investigation revealed remarkable findings regarding the relationship between stress and student performance. Analysis of the descriptive statistics and means of each group suggests that low performers have increased stress and perform poorly, whereas high performers have increased stress and perform superbly, and moderate performers have modest stress and perform moderately. PMID:20879631

McKay, Kelly A Chiffer; Buen, John E; Bohan, Kevin J; Maye, John P

2010-08-01

127

May Salivary Alpha-Amylase Level Be a Useful Tool for Assessment of the Severity of Schizophrenia and Evaluation of Therapy? A Case Report  

PubMed Central

Background. Previous studies suggested dysfunction of the autonomic nervous system (ANS) in schizophrenia patients, but the mechanism remains unclear. Recently, the measurement of salivary alpha-amylase (sAA) has been considered a useful tool for evaluating ANS, especially the sympathoadrenal medullary system. Furthermore, there was a report that patients with schizophrenia showed much higher sAA level than normal controls. Methods. We present the case of a 51-year-old female with catatonic schizophrenia. She needed the treatment of electroconvulsive therapy (ECT). We evaluated her sAA level and her psychiatric symptoms during the treatment. Results. Before ECT treatment, she showed high sAA level. Her sAA level decreased during the course of ECT, and this attenuation was accompanied by improvement of schizophrenic symptoms. Conclusion. We consider that measurement of the sAA level may be one of the useful biological markers for assessment of psychotic state and efficacy of treatment in patients with schizophrenia.

Ieda, Masa; Miyaoka, Tsuyoshi; Kawano, Kiminori; Wake, Rei; Inagaki, Takuji; Horiguchi, Jun

2012-01-01

128

Improved Chromogenic Substrate for Determination of Amylase Activity.  

National Technical Information Service (NTIS)

The patent application discloses an improved chromogenic substrate for alpha-amylase assays. The substrate synthesized by reacting amylose with Cibachron Blue F3GA, sodium sulfate and trisodium phosphate and incubating the resultant dyed amylose substrate...

T. M. Dougherty

1976-01-01

129

Insoluble but enzymatically active ? -amylase from Bacillus licheniformis  

Microsoft Academic Search

The gene encoding thermostable ?-amylase from Bacillus licheniformis consisting of 483 amino acid residues (mature protein) was cloned and expressed in Escherichia coli under the control of T7 promoter. The analysis of the soluble and insoluble fractions after lyzing the host cells revealed\\u000a that recombinant ?-amylase was produced in insoluble aggregates. Despite being produced in the insoluble aggregates the recombinant

Naeem Rashid; Alia Farooq; Muhammad Akhtar

2009-01-01

130

Interaction of different polyphenols with bovine serum albumin (BSA) and human salivary alpha-amylase (HSA) by fluorescence quenching.  

PubMed

Phenolic compounds are responsible for major organoleptic characteristics of plant-derived food and beverages; these substances have received much attention, given that the major function of these compounds is their antioxidant ability. In the context of this study, our major aim was study the binding of several phenolic compounds such as (+)-catechin, (-)-epicatechin, (-)-epicatechin gallate, malvidin-3-glucoside, tannic acid, procyanidin B4, procyanidin B2 gallate, and procyanidin oligomers to different proteins (bovine serum albumin and human alpha-amylase) by fluorescence quenching of protein intrinsic fluorescence. From the spectra obtained, the Stern-Volmer, the apparent static, and the bimolecular quenching constants were calculated. The structure of polyphenols revealed to significantly affect the binding/quenching process; in general, the binding affinity increased with the molecular weight of polyphenol compounds and in the presence of galloyl groups. For catechin monomer and procyanidin dimer B4, the K(SV) was 14,100 and 13,800 M(-1), respectively, and for galloyl derivatives, the K(SV) was 19,500 and 21,900 M(-1), respectively. Tannic acid was shown to be the major quenching molecule for both proteins. However, comparing different proteins, the same polyphenol showed different quenching effects, which are suggested to be related to the three-dimensional structure of the proteins studied. For (+)-catechin and BSA, the K(SV) was 8700 M(-1), and with alpha-amylase, it was 14,100 M(-1); for tannic acid, the K(SV) was 10,0548 and 11,0674 M(-1), respectively. From the results obtained, besides the main binding analysis performed, we conclude that this technique is more sensitive than thought because we can detect several interactions that have not been proven by other methods, namely, nephelometry. Overall, fluorescence quenching has proven to be a very sensitive technique with many potentialities to analyze the interaction between polyphenols and proteins. PMID:17636939

Soares, Susana; Mateus, Nuno; Freitas, Victor de

2007-08-01

131

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

Microsoft Academic Search

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 dipep- tidases. The aim of this study was to verify the peptidase activity of AbpB and

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

2008-01-01

132

Constituents of stem bark of Callistemon rigidus showing inhibitory effects on mouse alpha-amylase activity.  

PubMed

From stem bark of Callistemon rigidus (Myrtaceae), piceatannol and scirpusin B were isolated as components that exhibit inhibitory effects on alpha-amylase activity in isolated mouse plasma. In particular, scirpusin B also inhibited alpha-amylase in mouse gastrointestinal tract. Thus, we expect the depressive effect on the elevation of postprandial blood glucose may be a new medicinal use of this compound as well as the plant itself. PMID:16755033

Kobayashi, Kyoko; Ishihara, Tamaki; Khono, Eriko; Miyase, Toshio; Yoshizaki, Fumihiko

2006-06-01

133

Rap1 Activation Plays a Regulatory Role in Pancreatic Amylase Secretion*S?  

PubMed Central

Rap1 is a member of the Ras superfamily of small GTP-binding proteins and is localized on pancreatic zymogen granules. The current study was designed to determine whether GTP-Rap1 is involved in the regulation of amylase secretion. Rap1A/B and the two Rap1 guanine nucleotide exchange factors, Epac1 and CalDAG-GEF III, were identified in mouse pancreatic acini. A fraction of both Rap1 and Epac1 colocalized with amylase in zymogen granules, but only Rap1 was integral to the zymogen granule membranes. Stimulation with cholecystokinin (CCK), carbachol, and vasoactive intestinal peptide all induced Rap1 activation, as did calcium ionophore A23187, phorbol ester, forskolin, 8-bromo-cyclic AMP, and the Epac-specific cAMP analog 8-pCPT-2?-O-Me-cAMP. The phospholipase C inhibitor U-73122 abolished carbachol- but not forskolin-induced Rap1 activation. Co-stimulation with carbachol and 8-pCPT-2?-O-Me-cAMP led to an additive effect on Rap1 activation, whereas a synergistic effect was seen on amylase release. Although the protein kinase A inhibitor H-89 abolished forskolin-stimulated CREB phosphorylation, it did not modify forskolin-induced GTP-Rap1 levels, excluding PKA participation. Overexpression of Rap1 GTPase-activating protein, which blocked Rap1 activation, reduced the effect of 8-bromo-cyclic AMP, 8-pCPT-2?-O-Me-cAMP, and vasoactive intestinal peptide on amylase release by 60% and reduced CCK- as well as carbachol-stimulated pancreatic amylase release by 40%. These findings indicate that GTP-Rap1 is required for pancreatic amylase release. Rap1 activation not only mediates the cAMP-evoked response via Epac1 but is also involved in CCK- and carbachol-induced amylase release, with their action most likely mediated by CalDAG-GEF III.

Sabbatini, Maria E.; Chen, Xuequn; Ernst, Stephen A.; Williams, John A.

2008-01-01

134

Potent ?-amylase inhibitory activity of Indian Ayurvedic medicinal plants  

Microsoft Academic Search

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

Sudha P; Smita S Zinjarde; Shobha Y Bhargava; Ameeta R Kumar

2011-01-01

135

Anticoagulant activities in salivary glands of tabanid flies.  

PubMed

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

Kazimírová, M; Sulanová, M; Trimnellt, A R; Kozánek, M; Vidlicka, L; Labuda, M; Nuttall, P A

2002-09-01

136

Phlebotomus papatasi and Leishmania major parasites express alpha-amylase and alpha-glucosidase.  

PubMed

Alpha-amylase and alpha-glucosidase activities were found in homogenates of young, unfed male and female Phlebotomus papatasi and in gut and salivary gland preparations. A significant increase in both enzyme activities in females and of alpha-amylase in males was recorded for flies that had fed overnight on a plant (Capparis spinosa). After plant feeding, alpha-amylase activity was relatively lower in female salivary glands and higher in guts, while in the males the activity in the salivary glands had increased. Alpha-glucosidase activity increased in guts of both sexes and in the salivary glands of the females. In addition, alpha-amylase activity was found in preparations of Leishmania major and L. infantum promastigotes, but not in those of L. donovani or L. tropica. Alpha-glucosidase activity was present in promastigote preparations of L. major, L. infantum, L. donovani, L. braziliensis, Crithidia fasciculata and Herpetomonas muscarum. It was lacking in similar preparations of L. tropica, Sauroleishmania agamae or Leptomonas seymouri. The growth rate of L. major promastigotes in medium supplemented with starch or with glucose was similar and it was significantly higher than in glucose poor medium. In this study, we demonstrate that P. papatasi and L. major possess the enzymes for hydrolyzing starch grains that are included in the plant tissue-diet of the sand flies. PMID:11164750

Jacobson, R L; Schlein, Y

2001-01-15

137

?-AMYLASE4, a Noncatalytic Protein Required for Starch Breakdown, Acts Upstream of Three Active ?-Amylases in Arabidopsis Chloroplasts[W][OA  

PubMed Central

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.

Fulton, Daniel C.; Stettler, Michaela; Mettler, Tabea; Vaughan, Cara K.; Li, Jing; Francisco, Perigio; Gil, Manuel; Reinhold, Heike; Eicke, Simona; Messerli, Gaelle; Dorken, Gary; Halliday, Karen; Smith, Alison M.; Smith, Steven M.; Zeeman, Samuel C.

2008-01-01

138

Directed evolution of a bacterial ?-amylase: Toward enhanced pH-performance and higher specific activity  

PubMed Central

?-Amylases, in particular, microbial ?-amylases, are widely used in industrial processes such as starch liquefaction and pulp processes, and more recently in detergency. Due to the need for ?-amylases with high specific activity and activity at alkaline pH, which are critical parameters, for example, for the use in detergents, we have enhanced the ?-amylase from Bacillus amyloliquefaciens (BAA). The genes coding for the wild-type BAA and the mutants BAA S201N and BAA N297D were subjected to error-prone PCR and gene shuffling. For the screening of mutants we developed a novel, reliable assay suitable for high throughput screening based on the Phadebas assay. One mutant (BAA 42) has an optimal activity at pH 7, corresponding to a shift of one pH unit compared to the wild type. BAA 42 is active over a broader pH range than the wild type, resulting in a 5-fold higher activity at pH 10. In addition, the activity in periplasmic extracts and the specific activity increased 4- and 1.5-fold, respectively. Another mutant (BAA 29) possesses a wild-type-like pH profile but possesses a 40-fold higher activity in periplasmic extracts and a 9-fold higher specific activity. The comparison of the amino acid sequences of these two mutants with other homologous microbial ?-amylases revealed the mutation of the highly conserved residues W194R, S197P, and A230V. In addition, three further mutations were found K406R, N414S, and E356D, the latter being present in other bacterial ?-amylases.

Bessler, Cornelius; Schmitt, Jutta; Maurer, Karl-Heinz; Schmid, Rolf D.

2003-01-01

139

Inhibitory effect of Gymnema Montanum leaves on ?-glucosidase activity and ?-amylase activity and their relationship with polyphenolic content  

Microsoft Academic Search

The present study was attempted to investigate the effect of G. montanum leaf extract on inhibition of ?-glucosidase and ?-amylase activity. The ethanol extract of G. montanum (GLEt) at various concentrations (1–10 ?g\\/ml) was tested for its inhibition pattern against ?-glucosidase and ?-amylase activity\\u000a in vitro and compared with the commercially available ?-glucosidase inhibitor, acarbose. The GLEt showed competitive inhibition\\u000a against

Kunga Mohan Ramkumar; Balsamy Thayumanavan; Thayumanavan Palvannan; Palanisamy Rajaguru

2010-01-01

140

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

Microsoft Academic Search

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

Surasak Laloknam; Supaporn Sirisopana; Somkiat Phornphisutthimas

2009-01-01

141

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

Microsoft Academic Search

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

M. C Hidalgo; E Urea; A Sanz

1999-01-01

142

Arginine is essential for the alpha-amylase inhibitory activity of the alpha-amylase/subtilisin inhibitor (BASI) from barley seeds.  

PubMed Central

Treatment of barley alpha-amylase/subtilisin inhibitor (BASI) with reagents specific for arginine, histidine, methionine and tyrosine residues and amino and carboxyl groups indicates that an arginine residue(s) is essential for its action on the target enzyme barley alpha-amylase 2. Phenylglyoxal modified eight out of 12 arginine residues in BASI. Kinetic analysis shows that the inactivation of BASI follows a pseudo-first-order reaction and is due to reaction with one molecule of phenylglyoxal; the second-order rate constant is determined to be 2.95 M-1.min-1. At pH 8.0, BASI and barley alpha-amylase 2 form an inactive 1:1 complex. The Ki value of this association is 2.2 x 10(-10) M. The alpha-amylase protects four arginine residues and also the alpha-amylase inhibitory activity of BASI against phenylglyoxal. When BASI from the phenylglyoxal-modified target enzyme-inhibitor complex is isolated and subjected to a second treatment with phenylglyoxal, four additional arginine residues are modified, with concomitant loss of the inhibitory activity. These results are discussed in relation to a three-dimensional model of BASI based on the known structure of the corresponding inhibitor from wheat.

Abe, J; Sidenius, U; Svensson, B

1993-01-01

143

Tissue specific expression of mouse alpha-amylase genes.  

PubMed

Two distinct alpha-amylase genes, Amy-1a and Amy-2a, are expressed in the mouse strain A/J. Amy-1a and Amy-2a are interrupted by 10 and 9 introns, respectively. With the exception of the first Amy-1a intron, which has no counterpart in Amy-2a, introns are located at analogous positions within the two genes. Comparable exons of Amy-1a and Amy-2a are more highly conserved in sequence than analogous introns. Amy-2a specifies pancreatic alpha-amylase mRNA. Two apparently identical copies of this gene exist in the haploid mouse genome. The single copy of Amy-1a is expressed in a tissue specific fashion in the salivary gland and the liver. It specifies alpha-amylase mRNA with identical translated and 3' nontranslated sequences but different 5' nontranslated sequences in the two tissues. These different mRNAs are generated by tissue specific splicing events. S1 nuclease mapping of nuclear transcripts from salivary gland and liver suggests the presence of at least two promotors of different strength in Amy-1a. A strong promoter appears to be active in the salivary gland exclusively, while a weak promoter is apparently used in both the salivary gland and the liver. The data suggest that regulation of Amy-1a expression occurs primarily at the transcriptional level. PMID:6186131

Schibler, U; Hagenbüchle, O; Young, R A; Tosi, M; Wellauer, P K

1982-01-01

144

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

PubMed

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

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

2014-11-15

145

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

PubMed

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

Matsui, Takemi; Katayose, Satoshi

2014-08-01

146

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

PubMed Central

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

Kumar, Ashok; Khan, Saleemulla

2011-01-01

147

?-Amylase Activity in Developing Sorghum Caryopses from Sprouting Resistant and Susceptible Varieties. The Role of ABA and GAs on its Regulation  

Microsoft Academic Search

?-Amylase activity was analysed in developing caryopses fromSorghumvarieties showing contrasting susceptibility to preharvest sprouting. Unsprouted caryopses from variety Redland B2 (very susceptible) showed ?-amylase activity from 31 d after pollination (DAP) onwards, that were 20-fold higher than those observed for variety IS 9530 (very resistant). When the caryopses were incubated the ?-amylase activity predominately increased in the embryos. IS 9530

EDUARDO A PAGANO; ROBERTO L BENECH-ARNOLD; MARISA WAWRZKIEWICZ; HAYDEE S STEINBACH

1997-01-01

148

Retention of enzymatic activity of alpha-amylase in the reductive synthesis of gold nanoparticles.  

PubMed

In this paper, we report the generation of Au nanoparticles (NPs), using a pure enzyme for the reduction of AuCl4(-), with the retention of enzymatic activity in the complex. As a model system, alpha-amylase was used to readily synthesize and stabilize Au NPs in aqueous solution. Although several other enzymes were also pursued for the synthesis, it was interesting to observe that only alpha-amylase and EcoRI could produce Au NPs. Following NP synthesis, the activity of the enzyme was retained in the Au NP-alpha-amylase complex. The presence of Au NPs and alpha-amylase in the complex was established by UV-visible and FT-IR spectroscopy, X-ray diffraction (XRD) and transmission electron microscopic (TEM) measurements. Our observations suggest that the presence of free and exposed S-H groups is essential in the reduction of AuCl4(-) to Au NPs. Structural analysis of the enzymes showed that both alpha-amylase and EcoRI enzymes have free and exposed S-H groups in their native form and thus are suitable for the generation of NPs, whereas the other ones used here do not have such groups. Fortuitously, the enzymatic functional group of alpha-amylase is positioned opposite to that of the free and exposed S-H group, which makes it ideal for the production of Au NPs; binding of the enzyme to Au NPs via Au-S bond and also retention of the biological activity of the enzyme. PMID:17425338

Rangnekar, Abhijit; Sarma, Tridib Kumar; Singh, Atul Kumar; Deka, Jashmini; Ramesh, Aiyagari; Chattopadhyay, Arun

2007-05-01

149

Biochemical characterization of the alpha-amylase inhibitor in mungbeans and its application in inhibiting the growth of Callosobruchus maculatus.  

PubMed

The insect Callosobruchus maculatus causes considerable damage to harvested mungbean seeds every year, which leads to commercial losses. However, recent studies have revealed that mungbean seeds contain alpha-amylase inhibitors that can inhibit the protein C. maculatus, preventing growth and development of the insect larvae in the seed, thus preventing further damage. For this reason, the use of alpha-amylase inhibitors to interfere with the pest's digestion process has become an interesting alternative biocontrolling agent. In this study, we have isolated and purified the alpha-amylase inhibitor from mungbean seeds (KPS1) using ammonium sulfate precipitation, gel filtration chromatography and reversed phase HPLC. We found that the alpha-amylase inhibitor, isolated as a monomer, had a molecular weight of 27 kDa. The alpha-amylase inhibitor was purified 750-fold with a final yield of 0.4 mg of protein per 30 g of mungbean seeds. Its specific activity was determined at 14.5 U (mg of protein)(-1). Interestingly, we found that the isolated alpha-amylase inhibitor inhibits C. maculatus alpha-amylase but not human salivary alpha-amylase. After preincubation of the enzyme with the inhibitor, the mungbean alpha-amylase inhibitor inhibited C. maculatus alpha-amylase activity by decreasing V(max) while increasing the K(m) constant, indicating that the mungbean alpha-amylase is a mix noncompetitive inhibitor. The in vivo effect of alpha-amylase inhibitor on the mortality of C. maculatus shows that the alpha-amylase inhibitor acts on C. maculatus during the development stage, by reducing carbohydrate digestion necessary for growth and development, rather than during the end laying/hatching stage. Our results suggest that mungbean alpha-amylase inhibitor could be a useful future biocontrolling agent. PMID:20099823

Wisessing, Anussorn; Engkagul, Arunee; Wongpiyasatid, Arunee; Choowongkomon, Kiattawee

2010-02-24

150

Comparison of Salivary Peroxidase System Components in Caries-Free and Caries-Active Naval Recruits.  

National Technical Information Service (NTIS)

Measurements of flow rate, pH, hypothiocyanite, thiocyanate and salivary peroxidase activity were made for unstimulated (drooled) whole saliva samples from 29 caries-free and 29 caries-active naval recruits. No significant differences were found between t...

B. L. Lamberts K. M. Pruitt E. D. Pederson M. P. Golding

1984-01-01

151

Gibberellic acid effects on germination and ?-amylase activity of winter wheats  

Microsoft Academic Search

Sensitivity to GA in non-Gai genome winter wheat (Triticum aestivum L.) cultivars was investigated to determine magnitude of variation of the trait, its association with other traits, and effects of geographical location of production. a-Amylase enzyme activity was measured before and after treatment with gibberellic acid in 18 cultivars grown at one location and in five cultivars grown at six

A. J. McCrate; M. T. Nielsen; G. M. Paulsen; E. G. Heyne

1981-01-01

152

Changes of serum amylase, its isozyme fractions and amylase-creatinine clearance ratio in dogs with experimentally induced acute pancreatitis.  

PubMed

To investigate the diagnostic application of amylase to canine pancreatic diseases, serum amylase activities, its isozyme fractions and amylase-creatinine clearance ratio (ACCR) were analyzed in normal intact dogs and dogs experimentally induced acute pancreatitis. There was no statistic difference between normal male and female dogs. Amylase specific activities in pancreatic tissue extracts were more than 2,300 times higher than that in serum, and were also higher than those in other tissues; parotid and mandibular salivary glands, lung, heart, liver, spleen, duodenum, jejunum, ileum and kidney. Following the chloroform injection into the pancreatic tissue, WBC increased from 6 to 240 hr and serum glucose significantly increased at 72 and 96 hr, and no urine glucose was detected. BUN as well as serum and urine creatinine showed normal levels. ACCR increased until 96 hr without statistic significance. Serum amylase activities increased significantly after 3 hr and its isozyme was separated into 4 fractions (Amy1-Amy4) in contrast to 3 fractions (Amy2-Amy4) in intact dogs. Since this extra Amy1 seen from 1 hr increasing after 6 hr similarly to other 3 fractions, the evaluation of serum amylase and its isozyme fractions was indicated to be useful for the diagnosis of acute pancreatitis in dogs. PMID:7521216

Akuzawa, M; Morizono, M; Nagata, K; Hayano, S; Sakamoto, H; Yasuda, N; Okamoto, K; Kawasaki, Y; Deguchi, E

1994-04-01

153

Streptococci and activities of sucrases and alpha-amylases in supragingival dental plaque and saliva in three caries activity groups.  

PubMed

Thirty-eight young adults participated in the study. They were divided in a caries-inactive group, a low caries activity group and a moderate to high caries activity group. Total cultivable bacteria, Streptococcus salivarius, and S. mutans in plaque and saliva were quantitated on TSA, MS, and MSB plates, respectively. Sucrase activity was determined by measuring reducing sugars in plaque and saliva after incubation with sucrose. alpha-Amylase activity was determined by Pharmacia Phadebas Amylase test. The data were analyzed with the non-parametric Mann-Whitney U test. The only significant difference was observed for plaque alpha-amylase activity between the caries-inactive group and the moderate-high caries activity group (P less than 0.05). The lack of differences concerning the other variables is discussed mainly on the basis of the multifactorial character of dental caries and the possible insufficiency of the applied methods. PMID:3485336

Fiehn, N E; Oram, V; Moe, D

1986-02-01

154

Salivary Gland Secretion.  

ERIC Educational Resources Information Center

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

Dorman, H. L.; And Others

1981-01-01

155

Continuous Production of Thermostable ?-Amylase with Clostridium thermosulfurogenes: Effect of Culture Conditions and Metabolite Levels on Enzyme Synthesis and Activity  

PubMed Central

A ?-amylase-overproducing mutant of Clostridium thermosulfurogenes was grown in continuous culture on soluble starch to produce thermostable ?-amylase. Enzyme productivity was reasonably stable over periods of weeks to months. The pH and temperature optima for ?-amylase production were pH 6.0 and 60°C, respectively. Enzyme concentration was maximized by increasing biomass concentration by using high substrate concentrations and by maintaining a low growth rate. ?-Amylase concentration reached 90 U ml?1 at a dilution rate of 0.07 h?1 in a 3% starch medium. A further increase in enzyme activity levels was limited by acetic acid inhibition of growth and low ?-amylase productivity at low growth rates.

Nipkow, A.; Shen, G.-J.; Zeikus, J. G.

1989-01-01

156

The PVA solution structure-change effect for a-amylase specific activation  

Microsoft Academic Search

SummaryThe effect of the addition of poly(vinyl alcohol) (PVA) on the ?-amylase activity from Bacillus subtilis was investigated. About 500% of spiky activation on the reaction rate was observed in the presence of 0.25wt% PVA in the reaction medium. This activation was induced by the addition of the PVA, and was resulted from a structural-change of the solution due to

Takashi Takada; Toshihiro Hirai

2004-01-01

157

Improvement of Bacillus circulans beta-amylase activity attained using the ancestral mutation method.  

PubMed

Thermostabilization of enzymes is one of the greatest challenges of protein engineering. The ancestral mutation method, which introduces ancestral residues into a target enzyme, has previously been developed and used to improve the thermostabilities of thermophilic enzymes. Herein, we report a study that used the ancestral mutation method to improve the thermostability of Bacillus circulans beta-amylase, a mesophilic enzyme. A bacterial, common-ancestral beta-amylase sequence was inferred using a phylogenetic tree composed of higher plant and bacterial amylase sequences. Eighteen mutants containing ancestral residues were designed, expressed in Escherichia coli and purified. Several of these mutants were more thermostable than that of the wild-type amylase. Notably, one mutant had both greater activity and greater thermostability. The relationship between the extent to which the amino acid residues within 5 A of the mutation site were evolutionarily conserved and the extent to which thermostability was improved was examined. Apparently, it is necessary to conserve the residues surrounding an ancestral residue if thermostability is to be improved by the ancestral mutation method. PMID:20406825

Yamashiro, Kan; Yokobori, Shin-ichi; Koikeda, Satoshi; Yamagishi, Akihiko

2010-07-01

158

Determination of amylase activity and amylase isoenzymes in serum and urine using a solid phase blue starch substrate.  

PubMed

The alpha-amylase of serum and urine was determined in 40 healthy people using the modified "Phadebas Amylase Test". The original method was modified by decreasing the incubation volume to one millilitre and by adding to all urine speciemens a small amount of albumin in saline. The normal values obtained are slightly higher than those obtained by the original method. The amylase isoenzymes were likewise determined from the serum and urine of the same 40 healthy people. For separation were used electrophoresis on cellulose acetate and Phadebas tablets as substrate. These urine and serum isoamylases were investigated and compared. The distributions obtained deviate somewhat from ones reported earlier. The clinical usefulness of isoamylase is briefly discussed. PMID:1145111

Ojala, K; Harmoinen, A

1975-03-01

159

Activation of central ?(2)-adrenoceptors mediates salivary gland vasoconstriction.  

PubMed

OBJECTIVE: Peripheral treatment with the cholinergic agonist pilocarpine increases salivary gland blood flow and induces intense salivation that is reduced by the central injection of moxonidine (?(2)-adrenoceptors/imidazoline agonist). In the present study, we investigated the effects of the intracerebroventricular (i.c.v.) injection of pilocarpine alone or combined with moxonidine also injected i.c.v. On submandibular/sublingual gland (SSG) vascular resistance. In addition, the effects of these treatments on arterial pressure, heart rate and on mesenteric and hindlimb vascular resistance were also tested. DESIGN: Male Holtzman rats with stainless steel cannula implanted into lateral ventricle and anaesthetized with urethane+?-chloralose were used. RESULTS: Pilocarpine (500nmol/1?l) injected i.c.v. Reduced SSG vascular resistance and increased arterial pressure, heart rate and mesenteric vascular resistance. Contrary to pilocarpine alone, the combination of moxonidine (20nmol/1?l) and pilocarpine injected i.c.v. Increased SSG vascular resistance, an effect abolished by the pre-treatment with the ?(2)-adrenoceptor antagonist yohimbine (320nmol/2?l). The increase in arterial pressure, heart rate and mesenteric resistance was not modified by the combination of moxonidine and pilocarpine i.c.v. CONCLUSION: These results suggest that the activation of central ?(2)-adrenoceptors may oppose to the effects of central cholinergic receptor activation in the SSG vascular resistance. PMID:22818538

Moreira, Thiago S; Takakura, Ana C; Menani, José V; Colombari, Eduardo

2012-07-18

160

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

PubMed

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

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

2012-09-12

161

The effect of 4-aminopyrazolo(3, 4-d) pyrimidine on mouse plasma ?-amylase activity  

Microsoft Academic Search

Summary  A single dose of 4-aminopyrazolo(3,4-d) pyrimidine (4APP), an adenine analog, was orally administered at various concentrations\\u000a to groups of mice and after 24 h, the plasma-?-amylase activity and cholesterol levels had significantly decreased in a dose-dependent\\u000a manner. There was no change, however, in pancreatic superoxide dismutase activity or the lipid peroxide level. Histologically,\\u000a zymogen degranulation was found after a high

Takeshi Minami; Kensuke Natsui; Hirofurni Nakagawa; Yuko Okazaki

1993-01-01

162

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

PubMed

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

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

2012-06-01

163

MDM2 is required for suppression of apoptosis by activated Akt1 in salivary acinar cells.  

PubMed

Chronic damage to the salivary glands is a common side effect following head and neck irradiation. It is hypothesized that irreversible damage to the salivary glands occurs immediately after radiation; however, previous studies with rat models have not shown a causal role for apoptosis in radiation-induced injury. We report that etoposide and gamma irradiation induce apoptosis of salivary acinar cells from FVB control mice in vitro and in vivo; however, apoptosis is reduced in transgenic mice expressing a constitutively activated mutant of Akt1 (myr-Akt1). Expression of myr-Akt1 in the salivary glands results in a significant reduction in phosphorylation of p53 at serine(18), total p53 protein accumulation, and p21(WAF1) or Bax mRNA following etoposide or gamma irradiation of primary salivary acinar cells. The reduced level of p53 protein in myr-Akt1 salivary glands corresponds with an increase in MDM2 phosphorylation in vivo, suggesting that the Akt/MDM2/p53 pathway is responsible for suppression of apoptosis. Dominant-negative Akt blocked phosphorylation of MDM2 in salivary acinar cells from myr-Akt1 transgenic mice. Reduction of MDM2 levels in myr-Akt1 primary salivary acinar cells with small interfering RNA increases the levels of p53 protein and renders these cells susceptible to etoposide-induced apoptosis in spite of the presence of activated Akt1. These results indicate that MDM2 is a critical substrate of activated Akt1 in the suppression of p53-dependent apoptosis in vivo. PMID:16982679

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

2006-12-01

164

Rapid laser nephelometric determination of amylase activity in serum and urine.  

PubMed

We describe herein a rapid and sensitive laser nephelometric method for the determination of serum and urinary amylase activities. Our data showed that the change in relative light scattering (RLS) of an amylopectin substrate measured by a laser nephelometer related directly with amylolytic activity of amylase from 50 to 600 IU/L. Within-run variations at 293 and 769 IU/L sera showed CV's of 5.0% and 3.1%, respectively. Day-to-day variation for the same sera showed CV's of 7.2% and 4.7%, respectively. Correlation studies using the manual Phadebas dye-starch complex method and with the Roche amylochrome method showed correlation coefficients of 0.99 and 0.95, respectively. Using urine specimens, the correlation studies also showed a correlation coefficient of 0.98. These studies indicated that the proposed method was sensitive, fast, economical and easily adaptable to emergency and routine applications. PMID:1713257

Liu, T Z; Wei, J S

1991-03-01

165

Role of adipokinetic hormone in stimulation of salivary gland activities: the fire bug Pyrrhocoris apterus L. (Heteroptera) as a model species.  

PubMed

The effect of adipokinetic hormone (Pyrap-AKH) in stimulating the function of insect salivary glands (SGs) in extra-oral digestive processes was studied in the firebug, Pyrrhocoris apterus L. (Heteroptera). The analyses were performed on samples of SGs and extracts of linden seeds, a natural source of the bug's food. The SGs from 3-day old P. apterus females (when the food ingestion culminates), primarily contained polygalacturonase (PG) enzyme activity, whereas the level of lipase, peptidase, amylase and ?-glucosidase was negligible. The transcription of PG mRNA and enzymatic activity were significantly increased in SGs after Pyrap-AKH treatment. The piercing and sucking of linden seeds by the bugs stimulated the intrinsic enzymatic cocktail of seeds (lipase, peptidase, amylase, glucosidase), and moreover the activity of these enzymes was significantly enhanced when the seeds were fed on by the Pyrap-AKH treated bugs. Similarly, a significant increase in PG activity was recorded in linden seeds fed on by hormonally-treated bugs or when injected by SG extract from hormonally treated ones as compared to untreated controls. The mechanism of AKH action in SGs is unknown, but likely involves cAMP (and excludes cGMP) as a second messenger, since the content of this compound doubled in SGs after Pyrap-AKH treatment. This new and as yet undescribed function of AKH in SGs is compared with the effect of this hormone on digestive processes in the midgut elucidated earlier. PMID:24269343

Vinokurov, Konstantin; Bedná?ová, Andrea; Tom?ala, Aleš; Stašková, Tereza; Krishnan, Natraj; Kodrík, Dalibor

2014-01-01

166

Endogenous avidin-binding activity in epithelial cells of the ducts of human salivary glands.  

PubMed

Avidin-biotin (AB) systems are commonly employed to investigate salivary gland sections either in immunohistochemistry or in immunofluorescence techniques. We noted non-specific staining in the ductal epithelium of minor salivary gland (SG) sections from 5 Sjögren's syndrome (SS), 5 chronic sialoadenitis (CS), and one normal parotid gland (NP) when incubated with AB peroxidase complex. Inhibition of endogenous peroxidase did not prevent the staining while saturation of supposed biotin-like molecules in the tissue with added avidin led to the loss of this non-specific staining. These results demonstrate the presence of endogenous avidin binding activity (EABA) in the epithelial cells of salivary gland ducts. We suggest that the avidin and streptavidin systems should not be used for immunohistological examination of the ductal epithelium of salivary glands. PMID:8162641

Cauli, A; Yanni, G; Panayi, G S

1994-01-01

167

Relationship between QTLs for preharvest sprouting and alpha-amylase activity in rye grain  

Microsoft Academic Search

Preharvest sprouting (PHS) and high alpha-amylase activity (AA) negatively affect quality of rye grain. The objective of this\\u000a study was to reveal genetic relationship between PHS and AA by developing a consensus map of QTLs controlling each trait.\\u000a A method of composite interval mapping (CIM) was used to search for QTLs within the 541 × Ot1-3 and DS2 × RXL10 F2 mapping populations representing

Piotr Masoj?; Pawe? Milczarski

2009-01-01

168

Role of Streptococcus gordonii Amylase-Binding Protein A in Adhesion to Hydroxyapatite, Starch Metabolism, and Biofilm Formation  

Microsoft Academic Search

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

JEFFREY D. ROGERS; ROBERT J. PALMER; PAUL E. KOLENBRANDER; FRANK A. SCANNAPIECO

2001-01-01

169

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

PubMed

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

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

2012-07-01

170

alpha-Amylase activity by the Beckman reaction system and suppression by pyruvate.  

PubMed

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

Hanson, N Q; Yasmineh, W G

1979-07-01

171

Thiol-oxidation reduces the release of amylase induced by ?-adrenergic receptor activation in rat parotid acinar cells.  

PubMed

In parotid acinar cells, the activation of ?-adrenergic receptors induces the accumulation of intracellular cAMP, and consequently provokes the exocytotic release of amylase, a digestive enzyme. The cellular redox status plays a pivotal role in regulating various cellular functions. Cellular redox imbalance caused by the oxidation of cellular antioxidants, as a result of oxidative stress, induces significant biological damage. In this study, we examined the effects of diamide, a thiol-oxidizing reagent, on amylase release by rat parotid acinar cells. In cells treated with diamide, the formation of cAMP and the release of amylase induced by the ?-agonist isoproterenol (IPR) were partially reduced. The inhibitory effect of diamide on the IPR-induced release of amylase could be abrogated by reduced glutathione or dithiothreitol. Diamide had no effect on the amylase release induced by forskolin, an adenylate cyclase activator, or by mastoparan, a heterotrimeric GTPbinding protein activator. In cells treated with diamide, the binding affinity for [(3)H]DHA, but not the number of binding sites, was reduced. These results suggest that ?-adrenergic receptor function is reduced by thiol-oxidation, which inhibits amylase secretion by parotid acinar cells. PMID:21079359

Guo, Ming-Yu; Satoh, Keitaro; Qi, Bing; Narita, Takanori; Katsumata-Kato, Osamu; Matsuki-Fukushima, Miwako; Fujita-Yoshigaki, Junko; Sugiya, Hiroshi

2010-10-01

172

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

SciTech Connect

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

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

2012-05-01

173

Purification and characterization of a halophilic ?-amylase with increased activity in the presence of organic solvents from the moderately halophilic Nesterenkonia sp. strain F.  

PubMed

An extracellular halophilic ?-amylase was purified from Nesterenkonia sp. strain F using 80 % ethanol precipitation and Q-Sepharose anion exchange chromatography. The enzyme showed a single band with an apparent molecular weight of 110 kDa by SDS-PAGE. The amylase exhibited maximal activity at pH 7-7.5, being relatively stable at pH 6.5-7.5. Optimal temperature for the amylase activity and stability was 45 °C. The purified enzyme was highly active in the broad range of NaCl concentrations (0-4 M) with optimal activity at 0.25 M NaCl. The amylase was highly stable in the presence of 3-4 M NaCl. Amylase activity was not influenced by Ca˛?, Rb?, Li?, Cs?, Mg˛? and Hg˛?, whereas Feł?, Cu˛?, Zn˛? and Alł?) strongly inhibited the enzyme activity. The ?-amylase was inhibited by EDTA, but was not inhibited by PMSF and ?-mercaptoethanol. K(m) value of the amylase for soluble starch was 6.6 mg/ml. Amylolytic activity of the enzyme was enhanced not only by 20 % of water-immiscible organic solvents but also by acetone, ethanol and chloroform. Higher concentration (50 %) of the water-miscible organic solvents had no significant effect on the amylase activity. To the best of our knowledge, this is the first report on increased activity of a microbial ?-amylase in the presence of organic solvents. PMID:22592324

Shafiei, Mohammad; Ziaee, Abed-Ali; Amoozegar, Mohammad Ali

2012-07-01

174

[The functional activity of the minor salivary glands of the lips in allergic cheilitis].  

PubMed

Functional activity of minor salivary glands is studied in 289 children, 52 of these without signs of cheilitis and 237 with allergic cheilitis at different stages. Three variants of secretion are distinguished: fast, medium, and slow. The ratio of these variants changed in exacerbation of disease in comparison with that in health or remission. The activity of the glands depends on clinical manifestations of cheilitis. Changes in the activity of minor salivary glands determine the clinical picture, localization, and type of involvement in cheilitis. PMID:10368598

Obraztsov, Iu L; Gorbatova, A N

1999-01-01

175

Active secretion and protective effect of salivary nitrate against stress in human volunteers and rats  

PubMed Central

Up to 25% of the circulating nitrate in blood is actively taken up, concentrated, and secreted into saliva by the salivary glands. Salivary nitrate can be reduced to nitrite by the commensal bacteria in the oral cavity or stomach and then further converted to nitric oxide (NO) in vivo, which may play a role in gastric protection. However, whether salivary nitrate is actively secreted in human beings has not yet been determined. This study was designed to determine whether salivary nitrate is actively secreted in human beings as an acute stress response and what role salivary nitrate plays in stress-induced gastric injury. To observe salivary nitrate function under stress conditions, alteration of salivary nitrate and nitrite was analyzed among 22 healthy volunteers before and after a strong stress activity, jumping down from a platform at the height of 68m. A series of stress indexes was analyzed to monitor the stress situation. We found that both the concentration and the total amount of nitrate in mixed saliva were significantly increased in the human volunteers immediately after the jump, with an additional increase 1 h later (p < 0.01). Saliva nitrite reached a maximum immediately after the jump and was maintained 1 h later. To study the biological functions of salivary nitrate and nitrite in stress protection, we further carried out a water-immersion-restraint stress (WIRS) assay in male adult rats with bilateral parotid and submandibular duct ligature (BPSDL). Intragastric nitrate, nitrite, and NO; gastric mucosal blood flow; and gastric ulcer index (UI) were monitored and nitrate was administrated in drinking water to compensate for nitrate secretion in BPSDL animals. Significantly decreased levels of intragastric nitrate, nitrite, and NO and gastricmucosal blood flow were measured in BPSDL rats during the WIRS assay compared to sham control rats (p < 0.05). Recovery was observed in the BPSDL rats upon nitrate administration. The WIRS-induced UI was significantly higher in the BPSDL animals compared to controls, and nitrate administration rescued the WIRS-induced gastric injury in BPSDL rats. In conclusion, this study suggests that stress promotes salivary nitrate secretion and nitrite formation, which may play important roles in gastric protection against stress-induced injury via the nitrate-dependent NO pathway.

Jin, Luyuan; Qin, Lizheng; Xia, Dengsheng; Liu, Xibao; Fan, Zhipeng; Zhang, Chunmei; Gu, Liankun; He, Junqi; Ambudkar, Indu S.; Deng, Dajun; Wang, Songlin

2014-01-01

176

Evaluation of ten wild nigerian mushrooms for amylase and cellulase activities.  

PubMed

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

Jonathan, Segun Gbolagade; Adeoyo, Olusegun Richard

2011-06-01

177

Evaluation of Ten Wild Nigerian Mushrooms for Amylase and Cellulase Activities  

PubMed Central

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.

Adeoyo, Olusegun Richard

2011-01-01

178

Determination of Antioxidant Capacity, ?-Amylase and Lipase Inhibitory Activity of Crotalaria Juncea Linn In Vitro Inhibitory Activity of Crotalaria Juncea Linn.  

PubMed

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

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

2014-06-01

179

Identification of Anticoagulant Activities in Salivary Gland Extracts of Four Horsefly Species (Diptera, Tabanidae)  

Microsoft Academic Search

Anticoagulant activities against the extrinsic and intrinsic coagulation pathways were identified in salivary gland extracts (SGE) prepared from four tabanids (Hybomitra muehlfeldi, Tabanus autumnalis, Haematopota pluvialis, Heptatoma pellucens). All extracts prolonged human plasma clotting time in a dose-dependent manner and inhibited thrombin activity in the chromogenic substrate assay. Horsefly SGE did not inhibit factor Xa. Partial purification of SGE proteins

M. Kazimírová; M. Šulanová; M. Kozánek; M. Labuda; P. A. Nuttall

2001-01-01

180

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

PubMed

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

Singh, Kritika; Kayastha, Arvind M

2014-11-01

181

P2X7 receptor activation induces inflammatory responses in salivary gland epithelium  

PubMed Central

Inflammation of the salivary gland is a well-documented aspect of salivary gland dysfunction that occurs in Sjogren's syndrome (SS), an autoimmune disease, and in ?-radiation-induced injury during treatment of head and neck cancers. Extracellular nucleotides have gained recognition as key modulators of inflammation through activation of cell surface ionotropic and metabotropic receptors, although the contribution of extracellular nucleotides to salivary gland inflammation is not well understood. In vitro studies using submandibular gland (SMG) cell aggregates isolated from wild-type C57BL/6 mice indicate that treatment with ATP or the high affinity P2X7R agonist 3?-O-(4-benzoyl)benzoyl-ATP (BzATP) induces membrane blebbing and enhances caspase activity, responses that were absent in SMG cell aggregates isolated from mice lacking the P2X7R (P2X7R?/?). Additional studies with SMG cell aggregates indicate that activation of the P2X7R with ATP or BzATP stimulates the cleavage and release of ?-fodrin, a cytoskeletal protein thought to act as an autoantigen in the development of SS. In vivo administration of BzATP to ligated SMG excretory ducts enhances immune cell infiltration into the gland and initiates apoptosis of salivary epithelial cells in wild-type, but not P2X7R?/?, mice. These findings indicate that activation of the P2X7R contributes to salivary gland inflammation in vivo, suggesting that the P2X7R may represent a novel target for the treatment of salivary gland dysfunction.

Woods, Lucas T.; Camden, Jean M.; Batek, Josef M.; Petris, Michael J.; Erb, Laurie

2012-01-01

182

The ram1 Mutant of Arabidopsis Exhibits Severely Decreased Amylase Activity1  

Microsoft Academic Search

Despite extensive biochemical analyses, the biological function(s) of plant -amylases remains unclear. The fact that -amylases degrade starch in vitro suggests that they may play a role in starch metabolism in vivo. -Amylases have also been suggested to prevent the accumulation of highly polymerized polysaccharides that might otherwise impede flux through phloem sieve pores. The identification and characterization of a

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

2001-01-01

183

Directed evolution of a bacterial alpha-amylase: toward enhanced pH-performance and higher specific activity.  

PubMed

alpha-Amylases, in particular, microbial alpha-amylases, are widely used in industrial processes such as starch liquefaction and pulp processes, and more recently in detergency. Due to the need for alpha-amylases with high specific activity and activity at alkaline pH, which are critical parameters, for example, for the use in detergents, we have enhanced the alpha-amylase from Bacillus amyloliquefaciens (BAA). The genes coding for the wild-type BAA and the mutants BAA S201N and BAA N297D were subjected to error-prone PCR and gene shuffling. For the screening of mutants we developed a novel, reliable assay suitable for high throughput screening based on the Phadebas assay. One mutant (BAA 42) has an optimal activity at pH 7, corresponding to a shift of one pH unit compared to the wild type. BAA 42 is active over a broader pH range than the wild type, resulting in a 5-fold higher activity at pH 10. In addition, the activity in periplasmic extracts and the specific activity increased 4- and 1.5-fold, respectively. Another mutant (BAA 29) possesses a wild-type-like pH profile but possesses a 40-fold higher activity in periplasmic extracts and a 9-fold higher specific activity. The comparison of the amino acid sequences of these two mutants with other homologous microbial alpha-amylases revealed the mutation of the highly conserved residues W194R, S197P, and A230V. In addition, three further mutations were found K406R, N414S, and E356D, the latter being present in other bacterial alpha-amylases. PMID:14500872

Bessler, Cornelius; Schmitt, Jutta; Maurer, Karl-Heinz; Schmid, Rolf D

2003-10-01

184

Anticoagulation activity of salivary gland extract of oriental blackfly Simulium indicum  

PubMed Central

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

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

2014-01-01

185

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

PubMed

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

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

2014-07-01

186

Berry polyphenols inhibit ?-amylase in vitro: identifying active components in rowanberry and raspberry.  

PubMed

Polyphenol-rich extracts from a range of berries inhibited ?-amylase in vitro, but the most effective were from raspberry and rowanberry (IC50 values of 21.0 and 4.5 ?g/mL, respectively). The inhibitory components were examined by different approaches. Extracts from yellow and red raspberries were equally able to inhibit ?-amylase. Because the yellow raspberry extracts effectively lacked anthocyanins, this suggested that they were not crucial for amylase inhibition. Notably, however, higher levels of other phenolic components in yellow raspberries (particularly, ellagitannins) did not increase amylase inhibition. Amylase inhibition in rowanberry was recovered in a fraction enriched in proanthocyanidins (PACs). Inhibition was ameliorated by bovine serum albumin, suggesting that PACs acted by binding to amylase. Co-incubation of rowanberry PACs with acarbose reduced the concentration of acarbose required for effective amylase inhibition. Such synergistic interactions could have implications for the current clinical use of acarbose for postprandial glycaemic control in type-2 diabetics. PMID:21329358

Grussu, Dominic; Stewart, Derek; McDougall, Gordon J

2011-03-23

187

Two Immunoregulatory Peptides with Antioxidant Activity from Tick Salivary Glands*  

PubMed Central

Ticks are blood-feeding arthropods that may secrete immunosuppressant molecules, which inhibit host inflammatory and immune responses and provide survival advantages to pathogens at tick bleeding sites in hosts. In the current work, two families of immunoregulatory peptides, hyalomin-A and -B, were first identified from salivary glands of hard tick Hyalomma asiaticum asiaticum. Three copies of hyalomin-A are encoded by an identical gene and released from the same protein precursor. Both hyalomin-A and -B can exert significant anti-inflammatory functions, either by directly inhibiting host secretion of inflammatory factors such as tumor necrosis factor-?, monocyte chemotectic protein-1, and interferon-? or by indirectly increasing the secretion of immunosuppressant cytokine of interleukin-10. Hyalomin-A and -B were both found to potently scavenge free radical in vitro in a rapid manner and inhibited adjuvant-induced inflammation in mouse models in vivo. The JNK/SAPK subgroup of the MAPK signaling pathway was involved in such immunoregulatory functions of hyalomin-A and -B. These results showed that immunoregulatory peptides of tick salivary glands suppress host inflammatory response by modulating cytokine secretion and detoxifying reactive oxygen species.

Wu, Jing; Wang, Yipeng; Liu, Han; Yang, Hailong; Ma, Dongying; Li, Jianxu; Li, Dongsheng; Lai, Ren; Yu, Haining

2010-01-01

188

Evidence for D1 Dopamine Receptor Activation by a Paracrine Signal of Dopamine in Tick Salivary Glands  

PubMed Central

Ticks that feed on vertebrate hosts use their salivary secretion, which contains various bioactive components, to manipulate the host's responses. The mechanisms controlling the tick salivary gland in this dynamic process are not well understood. We identified the tick D1 receptor activated by dopamine, a potent inducer of the salivary secretion of ticks. Temporal and spatial expression patterns examined by immunohistochemistry and reverse transcription polymerase chain reaction suggest that the dopamine produced in the basal cells of salivary gland acini is secreted into the lumen and activates the D1 receptors on the luminal surface of the cells lining the acini. Therefore, we propose a paracrine function of dopamine that is mediated by the D1 receptor in the salivary gland at an early phase of feeding. The molecular and pharmacological characterization of the D1 receptor in this study provides the foundation for understanding the functions of dopamine in the blood-feeding of ticks.

Simo, Ladislav; Koci, Juraj; Zitnan, Dusan; Park, Yoonseong

2011-01-01

189

Immunocytochemical Identification and Localization of Active and Inactive ?-Amylase and Pullulanase in Cells of Clostridium thermosulfurogenes EM1  

PubMed Central

Clostridium thermosulfurogenes EM1 formed blebs, i.e., protrusions still in contact with the cytoplasmic membrane, that originated from the cytoplasmic membrane during growth in batch culture and continuous culture. They could be observed squeezed between the cell wall and cytoplasmic membrane in cells with seemingly intact wall layers (surface layer and peptidoglycan layer) as well as in cells with wall layers in different states of degradation caused by phosphate limitation or high dilution rates. Blebs were found to turn into membrane vesicles by constriction in cases when the cell wall was heavily degraded. Bleb and vesicle formation was also observed in the absence of substrates that induce ?-amylase and pullulanase synthesis. No correlations existed between bleb formation and the presence of active enzyme. Similar blebs could also be observed in a number of other gram-positive bacteria not producing these enzymes, but they were not observed in gram-negative bacteria. For immunoelectron-microscopic localization of ?-amylase and pullulanase in C. thermosulfurogenes EM1, two different antisera were applied. One was raised against the enzymes isolated from the culture fluid; the other was produced against a peptide synthesized, as a defined epitope, in analogy to the N-terminal amino acid sequence (21 amino acids) of the native extracellular ?-amylase. By using these antisera, ?-amylase and pullulanase were localized at the cell periphery in samples taken from continuous culture or batch culture. In samples prepared for electron microscopy by freeze substitution followed by ultrathin sectioning, blebs could be seen, and the immunolabel pinpointing ?-amylase enzyme particles was seen not only randomly distributed in the cell periphery, but also lining the surface of the cytoplasmic membrane and the blebs. Cells exhibiting high or virtually no enzyme activity were labeled similarly with both antisera. This finding strongly suggests that ?-amylase and pullulanase may occur in both active and inactive forms, depending on growth conditions. Images

Specka, U.; Spreinat, A.; Antranikian, G.; Mayer, F.

1991-01-01

190

Amylase levels in semen and saliva stains.  

PubMed

Amylase levels were determined for 148 semen samples and 20 saliva samples as well as for their corresponding stains. The effect of aging on the detectability of amylase activity in these stains was also investigated. The Phadebas amylase test was used for the quantitative assay of amylase. High levels of amylase in fluid saliva resulted in high levels being detected in saliva stains. Lower levels present in most seminal fluids produce little or no detectable amounts of amylase in stains. Interpretations are made as to the possible sources of amylase activity found in stains from laboratory casework based on both the amylase concentration and the elapsed time between collection and analysis. The evidential value of the presence or absence of amylase activity in casework stains is also discussed. PMID:2423634

Auvdel, M J

1986-04-01

191

Perlecan Domain IV Peptide Stimulates Salivary Gland Cell Assembly In Vitro  

PubMed Central

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. ?-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 ?-amylase.

Pradhan, Swati; Zhang, Chu; Jia, Xinqiao; Carson, Daniel D.; Witt, Robert

2009-01-01

192

Inhibitory effect of Azadirachta indica A. juss leaf extract on the activities of alpha-amylase and alpha-glucosidase.  

PubMed

In recent decades, there has been a drastic increase in the incidence and prevalence of diabetic mellitus. The aim of this study was to evaluate the in vitro inhibitory effect of Azadirachita indica leaf extract on the activity of alpha-amylase and alpha-glucosidase as a means of alleviating hyperglycemia and managing diabetes mellitus. Aqueous extract of Azadirachita indica exhibited most potent alpha-amylase inhibitory activity with IC50 value of 9.15 mg mL(-1) and acetone extract exhibited most potent alpha-glucosidase inhibitory activity with IC50 value of 5.00 mg mL(-1). Kinetic studies revealed both acetone and aqueous extract to exhibit mixed non-competitive inhibition for alpha-amylase and alpha-glucosidase. It can be concluded that the antidiabetic potential of Azadirachta indica may be due to its ability to inhibit both alpha-amylase and alpha-glucosidase. The presence of phytochemicals such as flavonoids, tannins and saponins in this plant may be responsible for its inhibitory activity on the two enzymes studied. PMID:24511747

Kazeem, M I; Dansu, T V; Adeola, S A

2013-11-01

193

Involvement of cAMP Response Element-Binding Protein Activation in Salivary Secretion  

Microsoft Academic Search

Objective: Saliva secretion is mediated by cAMP and the calcium signaling pathway in salivary acinar cells. The PKA signaling pathway plays an important role in protein secretion through the activation of cAMP, in fluid secretion through the elevation of intracellular calcium and in the activation of cAMP response element-binding protein (CREB), which is involved in these signaling cascades. In this

Koichi Yamada; Hiroko Inoue; Satoshi Kida; Shoichi Masushige; Tatsuaki Nishiyama; Kenji Mishima; Ichiro Saito

2006-01-01

194

A proteomics approach to detect tissue-wide adaptive changes in the pancreas associated with increased pancreatic ?-amylase activity in domestic cattle (Bos taurus).  

PubMed

We used a proteomics-based approach to investigate potential regulatory proteins in the pancreas of domestic cattle (Bos taurus) that were associated with differences in pancreatic ?-amylase activity. Two groups of 48 and 45 crossbred steers in years 1 and 2, respectively, were fed a high moisture corn-based diet and were ranked according to their pancreatic ?-amylase activity. Steers (n=18) with high, medium, and low ?-amylase activity were selected, with 3 for each activity range and 9 for each experimental year, and their proteomic profiles were compared. Pancreatic samples from each animal were fractionated using 2D-HPLC and fractions detected using UV spectrophotometry. Software analysis revealed 119 common protein fractions among the 18 animals, and statistical analysis revealed 10 of these fractions differing (P<0.10) in abundance between animals from the high and low pancreatic ?-amylase activity groups. Five protein fractions identified after tandem mass spectrometry analysis and database searches were found to match proteins with protein-binding, nucleotide/DNA-binding or enzymatic capabilities. Bioinformatics analysis of these fractions revealed porphobilinogen deaminase, a DNA-binding protein, and a putative S1 peptidase that increased in abundance with increasing ?-amylase activity; with a putative ATP/GTP binding protein decreasing in abundance with increasing pancreatic ?-amylase activity. Changes in these fractions may represent adaptations of the pancreas in domestic cattle that are associated with differences in ?-amylase activity. PMID:23274691

Holligan, Simone; Wang, Jiaxi; Cant, John P; Swanson, Kendall C

2013-03-01

195

Aleurones from a Barley with Low [alpha]-Amylase Activity Become Highly Responsive to Gibberellin When Detached from the Starchy Endosperm.  

PubMed Central

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.

Skadsen, R. W.

1993-01-01

196

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

PubMed Central

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

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

2013-01-01

197

Genomic architecture of alpha-amylase activity in mature rye grain relative to that of preharvest sprouting.  

PubMed

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

Masoj?, Piotr; Wi?niewska, Magdalena; ?a?, Anna; Milczarski, Pawe?; Berdzik, Marcin; P?dziwiatr, Daniel; Pol-Szyszko, Magdalena; Ga??za, Monika; Owsianicki, Rados?aw

2011-05-01

198

A normal paediatric amylase range.  

PubMed

A normal paediatric range of plasma alpha-amylase activity was determined using the Phadebas blue starch method. The range for children over one year was 98--405 IU/l. Plasma amylase activity increased throughout infancy. Mature levels of activity were observed in some children by age 2 months and in most of them by 9 months. PMID:6155826

Aggett, P J; Taylor, F

1980-03-01

199

Effect of salivary pellicle on antibacterial activity of novel antibacterial dental adhesives using a dental plaque microcosm biofilm model  

PubMed Central

Objectives Antibacterial primer and adhesive are promising to inhibit biofilms and caries. Since restorations in vivo are exposed to saliva, one concern is the attenuation of antibacterial activity due to salivary pellicles. The objective of this study was to investigate the effects of salivary pellicles on bonding agents containing a new monomer dimethylaminododecyl methacrylate (DMADDM) or nanoparticles of silver (NAg) against biofilms for the first time. Methods DMADDM and NAg were synthesized and incorporated into Scotchbond Multi-Purpose adhesive and primer. Specimens were either coated or not coated with salivary pellicles. A microcosm biofilm model was used with mixed saliva from ten donors. Two types of culture medium were used: an artificial saliva medium (McBain), and Brain Heart Infusion (BHI) medium without salivary proteins. Metabolic activity, colony-forming units (CFU), and lactic acid production of plaque microcosm biofilms were measured (n = 6). Results Bonding agents containing DMADDM and NAg greatly inhibited biofilm activities, even with salivary pellicles. When using BHI, the pre-coating of salivary pellicles on resin surfaces significantly decreased the antibacterial effect (p < 0.05). When using artificial saliva medium, pre-coating of salivary pellicles on resin did not decrease the antibacterial effect. These results suggest that artificial saliva yielded medium-derived pellicles on resin surfaces, which provided attenuating effects on biofilms similar to salivary pellicles. Compared with the commercial control, the DMADDM-containing bonding agent reduced biofilm CFU by about two orders of magnitude. Significance Novel DMADDM- and NAg-containing bonding agents substantially reduced biofilm growth even with salivary pellicle coating on surfaces, indicating a promising usage in saliva-rich environment. DMADDM and NAg may be useful in a wide range of primers, adhesives and other restoratives to achieve antibacterial and anti-caries capabilities.

Li, Fang; Weir, Michael D.; Fouad, Ashraf F.; Xu, Hockin H.K.

2014-01-01

200

The two promoters of the mouse alpha-amylase gene Amy-1a are differentially activated during parotid gland differentiation.  

PubMed

Mouse parotid acinar cells differentiate and proliferate mainly after birth. During the first 3 weeks of age, alpha-amylase mRNA, one of the major gene products of the adult tissue, increases from barely detectable to adult levels (10(4) copies/cell). Run-on transcription experiments show that this increase is transcriptionally regulated. Northern blot hybridization and in situ hybridization results indicate that the two promoters of the alpha-amylase gene Amy-1a are differentially switched on. First, the weaker downstream promoter is activated, and by 2 weeks of age, virtually all acinar cells have accumulated the transcript initiated at this promoter. At this age the strong Amy-1a promoter is utilized in only a minor proportion of acinar cells, while in the adult this promoter appears to be active in all acinar cells. Thus, the progressive accumulation of alpha-amylase mRNA during postnatal parotid differentiation is mainly the consequence of progressive acinar cell commitment to expression of the strong parotid-specific Amy-1a promoter. The pattern of committing cells during differentiation suggests that once an acinar cell has initiated expression of parotid-type alpha-amylase mRNA, this commitment is passed on to its daughter cells. PMID:3872721

Shaw, P; Sordat, B; Schibler, U

1985-04-01

201

Secretion of mouse alpha-amylase from fission yeast Schizosaccharomyces pombe: presence of chymostatin-sensitive protease activity in the culture medium.  

PubMed

We have constructed two secretion vectors for Schizosaccharomyces pombe using an SV40 promoter and the secretion signals of the pGKL killer toxin complex derived from Kluyveromyces lactis. Although indigenous secretory glycoproteins tend to accumulate in the periplasmic space of S. pombe, we have succeeded in the secretion of mouse alpha-amylase into the culture medium. The efficiency of secretion, processing pattern, stability and culture conditions for mouse alpha-amylase were studied in S. pombe. The 128 kDa killer secretion signal was more effective in directing secretion of mouse alpha-amylase than the 28 kDa killer secretion signal. We detected a chymostatin-sensitive protease activity in the culture medium of S. pombe, which digests mouse alpha-amylase secreted into the culture medium. The addition of 5 micrograms/ml chymostatin was shown to protect mouse alpha-amylases from this degradation. PMID:8511968

Tokunaga, M; Kawamura, A; Yonekyu, S; Kishida, M; Hishinuma, F

1993-04-01

202

Neohesperidin dihydrochalcone: presentation of a small molecule activator of mammalian alpha-amylase as an allosteric effector.  

PubMed

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<1activator of the enzyme and based on the results obtained from modeling tools, it is suggested to interact with PPA at a hydrophilic site located at the N-terminal, far from the active site of the enzyme. PMID:23376024

Kashani-Amin, Elaheh; Larijani, Bagher; Ebrahim-Habibi, Azadeh

2013-03-18

203

Inhibitory Activities of Cyanidin and Its Glycosides and Synergistic Effect with Acarbose against Intestinal ?-Glucosidase and Pancreatic ?-Amylase  

PubMed Central

Cyanidin and its glycosides are naturally dietary pigments which have been indicated as promising candidates to have potential benefits to humans, especially in the prevention and treatment of diabetes mellitus. We investigated the structure activity relationships of cyanidin and its glycosides to inhibit intestinal ?-glucosidases and pancreatic ?-amylase in vitro. The results found that cyanidin and its glycosides are more specific inhibitors of intestinal sucrase than intestinal maltase. Cyanidin-3-galactoside and cyanidin-3-glucoside were the most potent inhibitors against intestinal sucrase and pancreatic ?-amylase with IC50 values of 0.50 ± 0.05 and 0.30 ± 0.01 mM, respectively. Our findings indicate that the structural difference between glucose and galactose at the 3-O-position of cyanidin was an important factor for modulating the inhibition of intestinal sucrase and pancreatic ?-amylase. The combination of cyandin-3-glucoside, cyanidin-3- galactoside or cyanidin-3,5-diglucosides with a low concentration of acarbose showed synergistic inhibition on intestinal maltase and sucrase. The synergistic inhibition was also found for a combination of cyanidin or cyanidin-3-glucoside with a low concentration of acarbose. The findings could provide a new insight into a use for the naturally occurring intestinal ?-glucosidase and pancreatic ?-amylase inhibitors for the prevention and treatment of diabetes and its complications.

Akkarachiyasit, Sarinya; Charoenlertkul, Piyawan; Yibchok-anun, Sirintorn; Adisakwattana, Sirichai

2010-01-01

204

Clinical evaluation of amylase-creatinine clearance ratio and amylase isoenzyme clearance in chronic renal failure.  

PubMed

Amylase-creatinine clearance ratio (ACCR) and amylase isoenzyme clearance were determined simultaneously in patients with chronic renal failure. ACCR in patients with compensated renal failure (3.5 +/- 0.4%) was not significantly different from normals (2.6 +/- 0.2%), while that in patients with non-compensated renal failure (6.7 +/- 0.4%) was significantly higher than that in normals. Clearance ratio of pancreatic isoamylase (Amylase-1) relative to creatinine clearance (CAmy . 1/Ccr) in patients with both compensated (5.9 +/- 1.0%) and non-compensated (6.8 +/- 0.4%) renal failure was as high as that in patients with acute pancreatitis (6.6 +/- 0.5%). On the other hand, clearance ratio of salivary isoamylase (Amylase-3) relative to creatinine clearance (CAmy . 3/CCr) in patients with compensated renal failure (1.5 +/- 0.3%) was almost the same as that in normals (2.1 +/- 0.1%), while that in patients with non-compensated renal failure was 5.9 +/- 0.7%, which was significantly higher than that in normals. The present study revealed that elevated ACCR in patients with severely impaired renal function was due to the increase of the clearance ratio for both pancreatic and salivary amylase. These facts suggested that glomerular permeability and tubular reabsorption for pancreatic and salivary amylase might play an important role on ACCR in patients with severely impaired renal function. PMID:6167484

Maeda, M; Otsuki, M; Okano, K; Yamasaki, T; Baba, S

1981-01-01

205

Determination of alpha-amylase activity in duodenal contents with the blue starch polymer.  

PubMed

The use of the blue starch polymer (Phadebas tablets) for the determination of duodenal alpha-amylase is described. 0.05% bovine or human albumin solutions are necessary for dissolution of substrate tablets as well as for dilution of duodenal contents. Concentration and output values of duodenal amylase before and after cholecystokinin-pancreozymin and secretin stimulation display a close correlation of the chromogenic and saccharogenic method. The average error of duodenal alpha-amylase in parallel examinations is low with both methods and their relative accuracy is practically the same. The chromogenic method is considered more simple and rapid. PMID:209677

Fric, P; Roth, Z

1977-01-01

206

The effectiveness of the Uchida-Kraepelin test for psychological stress: an analysis of plasma and salivary stress substances  

Microsoft Academic Search

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

Koreaki Sugimoto; Aya Kanai; Noriaki Shoji

2009-01-01

207

Amylase - urine  

MedlinePLUS

This test is done to diagnose pancreatitis and other diseases that affect the pancreas. ... amylase levels may be a sign of: Acute pancreatitis Alcohol consumption Cancer of the pancreas , ovaries, or ...

208

Identification of anticoagulant activities in salivary gland extracts of four horsefly species (Diptera, tabanidae).  

PubMed

Anticoagulant activities against the extrinsic and intrinsic coagulation pathways were identified in salivary gland extracts (SGE) prepared from four tabanids (Hybomitra muehlfeldi, Tabanus autumnalis, Haematopota pluvialis, Heptatoma pellucens). All extracts prolonged human plasma clotting time in a dose-dependent manner and inhibited thrombin activity in the chromogenic substrate assay. Horsefly SGE did not inhibit factor Xa. Partial purification of SGE proteins using reversed-phase high-performance liquid chromatography revealed species-specific differences in the elution profiles and range of fractions with anticoagulant activities. PMID:11910198

Kazimírová, M; Sulanová, M; Kozánek, M; Takác, P; Labuda, M; Nuttall, P A

2001-01-01

209

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

PubMed Central

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

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

2013-01-01

210

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

PubMed Central

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

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

2013-01-01

211

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

PubMed Central

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.

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

2013-01-01

212

The effects of Fusilade (Fluazifop- p -butyl) on germination, mitotic frequency and ?-amylase activity of lentil ( Lens culinaris Medik.) seeds  

Microsoft Academic Search

In this study, seed germination percentages, effects on phases of mitosis and ?-amylase enzyme activity of lentil seeds treated\\u000a with four different concentrations (0.25, 0.5, 1 and 1.5%) of Fusilade (Fluazifop-p-butyl) were determined. Median EC (effective concentration) values were calculated according to seed germination percentages\\u000a after treatment for 72 h. Germination percentages of primary lentil roots decreased with increasing Fusilade concentrations.

Feruzan Dane; Filiz Ekinci Sanal; Tulin Aktac

2007-01-01

213

The effect of ultrasound in combination with thermal treatment on the germinated barley’s alpha-amylase activity  

Microsoft Academic Search

The effects of ultrasound as emerging technology along with thermal treatment were investigated on the activity of barley’s\\u000a alpha-amylase after germination. All experiments were carried out at 20 kHz on an ultrasonic generator by considering the\\u000a three effective factors, temperature (30, 50 and 70°C) and ultrasonic intensities (20, 60 and 100% setting from total power\\u000a of device (460 W)) in

Maryam Yaldagard; Seyed. Ali. Mortazavi; Farideh. Tabatabaie

2008-01-01

214

Searching for Amylase  

NSDL National Science Digital Library

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

Keith D. Stanley (Beloit College;Biology); Ethel D. Stanley (Beloit College;Biology)

2006-05-20

215

Salivary cholinesterase activity in children with organic and convential diets  

EPA Science Inventory

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

216

The Diagnostic Potential of Salivary Protease Activities in Periodontal Health and Disease  

PubMed Central

Periodontal disease is characterised by proteolytic processes involving enzymes that are released by host immune cells and periodontal bacteria. These enzymes, when detectable in whole saliva, may serve as valuable diagnostic markers for disease states and progression. Because the substrate specificities of salivary proteases in periodontal health and disease are poorly characterised we probed these activities using several relevant substrates: 1) gelatin and collagen type IV; 2) the Arg/Lys–rich human salivary substrate histatin-5; and 3) a histatin-derived synthetic analog benzyloxycarbonyl-Arg-Gly-Tyr-Arg-methyl cumaryl amide (Z-RGYR-MCA). Substrate degradation was assessed in gel (zymography) and in solution. Whole saliva supernatant enzyme activities directed at gelatin, quantitated from the 42 kDa, 92 kDa and 130 kDa bands in the zymograms, were 1.3, 1.4 and 2.0 fold higher, respectively, in the periodontal patient group (p<0.01), consistent with enhanced activities observed towards collagen type IV. On the other hand, histatin 5 degraded equally fast in healthy and periodontal patients' whole saliva supernatant samples (p>0.10). Likewise, the hydrolysis rates of the Z-RGYR-MCA substrate were the same in the healthy and periodontal patient groups (p>0.10). In conclusion, gelatinolytic/collagenolytic activities but not trypsin-like activities in human saliva differentiate health from periodontal disease, and may thus provide an adjuvant to diagnosis for monitoring of disease activity.

Thomadaki, Konstantina; Bosch, Jos A.; Oppenheim, Frank G.; Helmerhorst, Eva J.

2013-01-01

217

The diagnostic potential of salivary protease activities in periodontal health and disease.  

PubMed

Periodontal disease is characterised by proteolytic processes involving enzymes that are released by host immune cells and periodontal bacteria. These enzymes, when detectable in whole saliva, may serve as valuable diagnostic markers for disease states and progression. Because the substrate specificities of salivary proteases in periodontal health and disease are poorly characterised, we probed these activities using several relevant substrates: (i) gelatin and collagen type IV; (ii) the Arg/Lys-rich human salivary substrate histatin-5; and (iii) a histatin-derived synthetic analog benzyloxycarbonyl-Arg-Gly-Tyr-Arg-methyl cumaryl amide (Z-RGYR-MCA). Substrate degradation was assessed in gel (zymography) and in solution. Whole saliva supernatant enzyme activities directed at gelatin, quantified from the 42 kDa, 92 kDa and 130 kDa bands in the zymograms, were 1.3, 1.4 and 2.0-fold higher, respectively, in the periodontal patient group (P < 0.01), consistent with enhanced activities observed towards collagen type IV. On the other hand, histatin 5 degraded equally fast in healthy and periodontal patients' whole saliva supernatant samples (P > 0.10). Likewise, the hydrolysis rates of the Z-RGYR-MCA substrate were the same in the healthy and periodontal patient groups (P > 0.10). In conclusion, gelatinolytic/collagenolytic activities but not trypsin-like activities in human saliva differentiate health from periodontal disease and may thus provide an adjuvant to diagnosis for monitoring disease activity. PMID:23379269

Thomadaki, K; Bosch, Ja; Oppenheim, Fg; Helmerhorst, Ej

2013-11-01

218

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

PubMed

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

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

2014-06-01

219

Wheat amylase trypsin inhibitors drive intestinal inflammation via activation of toll-like receptor 4.  

PubMed

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

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; Schuppan, Detlef

2012-12-17

220

Wheat amylase trypsin inhibitors drive intestinal inflammation via activation of toll-like receptor 4  

PubMed Central

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.

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.

2012-01-01

221

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

PubMed

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

2013-12-01

222

Salivary Diagnostics  

MedlinePLUS

... a clear, protein-rich fluid secreted by the salivary glands and trace amounts of various biochemicals present in ... genes and proteins that are expressed in the salivary glands. With these vast catalogues as their guide, they ...

223

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

PubMed Central

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

Olennikov, Daniil N.; Kashchenko, Nina I.

2014-01-01

224

Morphology and preliminary enzyme characterization of the salivary glands from the predatory bug Podisus nigrispinus (Heteroptera: Pentatomidae).  

PubMed

Podisus nigrispinus (Dallas) is a common predator in agricultural and natural systems in Neotropical America. Its feeding strategy involves extra-oral digestion and to better understand this process its salivary glands were extracted and subjected to morphological and preliminary enzyme characterization. The salivary glands of P. nigrispinus are formed by a pair of main and accessory gland complexes. The main salivary glands are further divided into an anterior and a posterior lobe. The compartmentalization of the salivary gland complex is likely to be important for the production, activation and release of the digestive enzymes used in the extra-oral digestion of prey items. Proteases and lipase, important digestive enzymes involved in zoophagy, were detected in the salivary glands of P. nigrispinus. The prevailing trypsin-like protease activity was characterized by using the serine-protease substrate N-alpha-benzoyl-L-Arg-p-nitroanilidine (L-BApNA) and the trypsin inhibitors tosyl-L-lysine chloromethyl ketone (TLCK) and benzamidine. The KM value obtained for trypsin-like activity was 1.57 mm and the different peaks of optimum pH and temperature activity suggest the presence of multiple forms of this enzyme in P. nigrispinus. Detection of amylase activity in the salivary glands of this predator suggests its ability to digest starch and obtain nutrients from plants, which may have adaptative value under prey scarcity. PMID:16768813

Oliveira, J A; Oliveira, M G A; Guedes, R N C; Soares, M J

2006-06-01

225

Effect of starch and amylase on the expression of amylase-binding protein A in Streptococcus gordonii.  

PubMed

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

Nikitkova, A E; Haase, E M; Scannapieco, F A

2012-08-01

226

Extra-Cellular Signal-Regulated ERK-1/ERK-2 Pathway Activation in Human Salivary Gland Mucoepidermoid Carcinoma  

PubMed Central

Information on oncogenetic events accompanying salivary gland mucoepidermoid carcinoma is so far limited. Activation of extracellular signal-regulated kinases ERK-1 and ERK-2 is strongly correlated to cancer. Using an antibody specific for phosphorylated (active) ERK-1/ERK-2, we examined human salivary gland mucoepidermoid carcinoma samples by immunohistochemistry. The comparison in paired tumor and normal tissue samples showed that phosphorylated ERK-1/ERK-2 immunoreactivity was higher in tumor cells as compared to surrounding normal salivary parenchyma. ERK-1/ERK-2 phosphorylation was observed in ?39% of mucoepidermoid carcinomas. Those tumors where the ERK-1/ERK-2 pathway was activated had a more aggressive tumor behavior as compared to the group where this pathway was inactive. The association of ERK-1/ERK-2 phosphorylation to a worse prognosis was independent of histological grade. ERK-1/ERK-2 phosphorylation was associated with increased Ki67 and cyclin A indexes, which indicated that ERK-1/ERK-2 pathway activation increased tumor cell proliferation. There was no relationship between ERK-1/ERK-2 phosphorylation and HER-2/neu or p16/INK4a protein expression. In conclusion, ERK-1/ERK-2 pathway is active in salivary gland mucoepidermoid carcinoma and this activation is associated to a more aggressive tumor behavior and a higher proliferative activity. These data suggest that deregulation of ERK-1/ERK-2 pathway contributes to mucoepidermoid carcinoma phenotype and, possibly, represents a target for new anticancer drugs.

Handra-Luca, Adriana; Bilal, Hadi; Bertrand, Jacques-Charles; Fouret, Pierre

2003-01-01

227

Expression of ?-Amylase from Alfalfa Taproots1  

PubMed Central

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

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

1998-01-01

228

AnAutomated, Saccharogenic Method for Determining SerumAmylase Activity  

Microsoft Academic Search

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

Wendell R. O'Neal

229

Asparagine as a nitrogen source for improving the secretion of mouse alpha-amylase in Saccharomyces cerevisiae protease A-deficient strains.  

PubMed

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

Chen, D C; Wang, B D; Chou, P Y; Kuo, T T

2000-02-01

230

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

PubMed Central

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

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

2014-01-01

231

Effect of Alcohol Withdrawl on Glutathione S-transferase, Total Antioxidant Capacity and Amylase in Blood and Saliva of Alcohol-Dependent Males  

PubMed Central

Introduction: Alcohol biomarkers help in the early detection of alcoholism and its complications. There is a paucity of studies in India on the salivary markers of systemic diseases in general and on salivary alcohol biomarkers in particular. Objectives: The present study was aimed at assessing the effect of alcohol withdrawal on the antioxidants and amylase in blood and saliva, and at finding the correlation between the blood and the salivary parameters in alcoholics. Methods: Sixty alcohol-dependent males who were in the age group of 30 – 70 years, who were admitted to the Deaddiction Centre for alcohol withdrawal treatment for one month, were the subjects of this study; age-matched healthy individuals were the controls. In the blood and saliva samples, the activities of Glutathione S-Transferase (GST) and amylase and the Total Antioxidant Capacity (TAC) were assayed. Results: The alcohol-dependent subjects showed significantly lower GST and amylase activities and the TAC in blood and saliva as compared to those in the controls (P<0.001). The alcohol withdrawal caused a significant increase in the GST and amylase activities and the TAC to near-control values. In the alcohol-dependent subjects, there was a significant correlation between the values in blood and saliva with respect to GST and TAC. Conclusions: Alcoholism causes an impaired antioxidant capacity and a decreased secretion of amylase, which is ameliorated due to the alcohol withdrawal regimen . The strong correlation between blood and saliva with respect to the antioxidants suggests the potential future use of saliva as a laboratory tool in clinical medicine.

Peter, Neethumol; Chiramel, Kevin J.; A.R., Shivashankara

2013-01-01

232

A single mouse alpha-amylase gene specifies two different tissue-specific mRNAs.  

PubMed

The alpha-amylase mRNAs which accumulate in two different tissues of the mouse, the salivary gland and the liver, are identical except for their 5' non-translated sequences: the 5' terminal 158 nucleotides of the major liver alpha-amylase mRNA are unrelated to the 5' terminal 47 nucleotides found in its salivary gland counterpart. DNA that specifies the 5'terminal one-quarter of these mRNAs has been isolated through genomic cloning and sequenced. The initial 161 nucleotides of the liver alpha-amylase mRNA are specified by DNA sequences that lie 4.5 kb upstream from those for the common body of the two mRNAs. In contrast, the 5' terminal 50 nucleotides of the salivary gland alpha-amylase mRNA are found 7.5 kb from sequences that the two mRNAs share in the genome. These cloned DNA sequences occur once per haploid genome, indicating that both the salivary gland and liver alpha-amylase mRNAs are transcribed from the same gene (Amy1A). Since no rearrangement of these DNA sequences can be detected among mouse sperm, salivary gland or liver preparations, gross rearrangement does not account for the tissue-specific pattern of expression observed for Amy1A. Rather, these data indicate that the salivary gland and liver alpha-amylase mRNAs are differentially transcribed and/or processed from identical DNA sequences in different tissues. PMID:6162570

Young, R A; Hagenbüchle, O; Schibler, U

1981-02-01

233

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

PubMed

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

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

2014-06-01

234

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

PubMed Central

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

2012-01-01

235

Effects of sorghum (Sorghum bicolor (L.) Moench) tannins on ?-amylase activity and in vitro digestibility of starch in raw and processed flours.  

PubMed

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

Mkandawire, Nyambe L; Kaufman, Rhett C; Bean, Scott R; Weller, Curtis L; Jackson, David S; Rose, Devin J

2013-05-01

236

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

NASA Astrophysics Data System (ADS)

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

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

2011-11-01

237

Tick salivary gland extract-activated transmission of Borrelia afzelii spirochaetes.  

PubMed

Saliva-activated transmission of Borrelia afzelii Canica, Nato, du Merle, Mazie, Baranton et Postic, 1993 was demonstrated using salivary gland extract (SGE) from Ixodes ricinus (L., 1758) ticks and C3H mice. Injection of Borrelia spirochaetes together with SGE increased the level of bacteraemia and accelerated the appearance of bacteria in the urinary bladder, compared with the injection of spirochaetes alone. More I. ricinus nymphs became infected when feeding on mice inoculated with B. afzelii plus SGE. Analysis of cytokines produced by cells of draining lymph nodes from SGE-treated mice showed a suppression of proinflammatory cytokines IFN-gamma, IL-6 and GM-CSF following a transient upregulation in comparison with the control mice infected without SGE. PMID:12194488

Pechová, Jitka; St?pánová, Gabriela; Kovár, Lubomír; Kopecký, Jan

2002-01-01

238

Galanin: hydrokinetic action on salivary glands in man.  

PubMed

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

Bauer, F E; Ghatei, M A; Zintel, A; Bloom, S R

1989-12-01

239

Voltage and Ca2+-activated K+ channel in baso-lateral acinar cell membranes of mammalian salivary glands  

NASA Astrophysics Data System (ADS)

Nervous or hormonal stimulation of many exocrine glands evokes release of cellular K+ (ref. 1), as originally demonstrated in mammalian salivary glands2,3, and is associated with a marked increase in membrane conductance1,4,5. We now demonstrate directly, by using the patch-clamp technique6, the existence of a K+ channel with a large conductance localized in the basolateral plasma membranes of mouse and rat salivary gland acinar cells. The K+ channel has a conductance of ~250 pS in the presence of high K+ solutions on both sides of the membrane. Although mammalian exocrine glands are believed not to possess voltage-activated channels1,7, the probability of opening the salivary gland K+ channel was increased by membrane depolarization. The frequency of channel opening, particularly at higher membrane potentials, was increased markedly by elevating the internal ionized Ca2+ concentration, as previously shown for high-conductance K+ channels from cells of neural origin8-10. The Ca2+ and voltage-activated K+ channel explains the marked cellular K+ release that is characteristically observed when salivary glands are stimulated to secrete.

Maruyama, Y.; Gallacher, D. V.; Petersen, O. H.

1983-04-01

240

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

NASA Astrophysics Data System (ADS)

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.

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

2012-03-01

241

Intracellular a-Amylase of Streptococcus mutans  

Microsoft Academic Search

Sequencing upstream of the Streptococcus mutans gene for a CcpA gene homolog, regM, revealed an open read- ing frame, named amy, with homology to genes encoding a-amylases. The deduced amino acid sequence showed a strong similarity (60% amino acid identity) to the intracellular a-amylase of Streptococcus bovis and, in com- mon with this enzyme, lacked a signal sequence. Amylase activity

CHRISTINE L. SIMPSON; ROY R. B. RUSSELL

1998-01-01

242

Inheritance of late maturity ?-amylase in wheat  

Microsoft Academic Search

Two wheat cultivars that consistently show high levels of grain a-amylase at harvest ripeness, in the absence of preharvest sprouting, were crossed with a control, low a-amylase cultivar, and F1, F2 and BC1 populations were developed. Grain of these populations was analysed for a-amylase activity at harvest ripeness. Distribution and segregation patterns were consistent with control at a single locus

Kolumbina Mrva; Daryl J. Mares

1996-01-01

243

Thermal stability and activity improvements of a Ca-independent ?-amylase from Bacillus subtilis CN7 by C-terminal truncation and hexahistidine-tag fusion.  

PubMed

Simultaneous improvements of thermostability and activity of a Ca-independent ?-amylase from Bacillus subtilis CN7 were achieved by C-terminal truncation and his6 -tag fusion. C-terminal truncation, which eliminates C-terminal 194-amino-acid residues from the intact mature ?-amylase, raised the turnover number by 35% and increased the thermostability in terms of half-life at 65 °C by threefold. A his6 -tag fusion at either the C- or N-terminus of truncated ?-amylase further enhanced its turnover number by 59% and 37%, respectively. Molecular modeling revealed that these improvements could be attributed to structural rearrangement and reorientation of the catalytic residues. PMID:24033784

Wang, Chenghua; Wang, Qingyan; Liao, Siming; He, Bingfang; Huang, Ribo

2014-03-01

244

An Interlaboratory Study of Amylase Methodologies Using Purified Enzyme Materials.  

National Technical Information Service (NTIS)

An interlaboratory study of amylase methodologies was conducted by the Center for Disease Control November 30, 1978. Participants in the study analyzed 7 samples for amylase activity -- 5 with a simulated serum matrix containing purified pancreatic amylas...

T. L. Hearn E. J. Sampson V. S. Whitner S. S. McKneally D. J. Boone

1978-01-01

245

A Comparative Study of Human Parotid and Submaxillary Saliva Amylase.  

National Technical Information Service (NTIS)

The amylase of human parotid and submaxillary gland secretions was isolated by chromatography on columns of Sephadex G-100. Acrylamide gel electrophoresis of the chromatographic fractions possessing amylase activity revealed matching isoenzyme stain patte...

B. L. Lamberts T. S. Meyer R. M. Osborne

1969-01-01

246

Therapeutic effects of soy isoflavones on ?-amylase activity, insulin deficiency, liver-kidney function and metabolic disorders in diabetic rats.  

PubMed

Natural estrogens have demonstrated a wide variety of biological activities, which makes them a good candidate for the treatment of diabetes. In vitro, this study evidenced that isoflavones enhanced insulin secretion and inhibited ?-amylase activity. In vivo, the findings indicated that soy isoflavones stimulated insulin secretion, increased the hepatic glycogen content and suppressed blood glucose level. The soy isoflavones were also protected hepatic-kidney functions showed by the significant increase in superoxide dismutase, catalase and glutathione peroxidase activities and the decrease in thiobarbituric acid reactive substances, total bilirubin, creatinine and transaminases content. Moreover, soy isoflavones induced a decrease in LDL-cholesterol and triglycerides and an increase in HDL-cholesterol in plasma and liver. Overall, the findings of the current study indicate that soy isoflavones exhibit attractive properties and can, therefore, be considered a promising candidate for future application as alternative therapeutic agents, particularly in the development of anti-diabetic and hypolipidaemic drugs. PMID:21108110

Hamden, Khaled; Jaouadi, Bassem; Carreau, Serge; Aouidet, Abdallah; Elfeki, Abdelfattah

2011-02-01

247

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

PubMed Central

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

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

2010-01-01

248

An assessment of four methods for the assay of amylase activity.  

PubMed

This paper describes an investigation carried out to select a reagent kit for amylase assay to replace the Wohlgemuth method currently in use in our laboratory. The following four commercial kits were assessed: Amylochrome, Phadebas, DyAmyl-L and Amylotube. The results showed that the first three kits which employed chromogenic substrates gave better reproducibility than the Amylotube kit which used plain starch substrate and the starch-iodine reaction. Results obtained with the former methods correlated well with one another but poorly with results obtained with the latter method. Various considerations given for the recommended use of the Amylochrome method are discussed. PMID:94982

Chua, K S; Tan, I K; Vengadiswaran, R; Peiris, J T

1979-04-01

249

Vicilin-like peptides from Capsicum baccatum L. seeds are ?-amylase inhibitors and exhibit antifungal activity against important yeasts in medical mycology.  

PubMed

The objective of this study was to isolate antimicrobial peptides from Capsicum baccatum seeds and evaluate their antimicrobial activity and inhibitory effects against ?-amylase. Initially, proteins from the flour of C. baccatum seeds were extracted in sodium phosphate buffer, pH 5.4, and precipitated with ammonium sulfate at 90% saturation. The D1 and D2 fractions were subjected to antifungal tests against the yeasts Saccharomyces cerevisiae, Candida albicans, Candida tropicalis, and Kluyveromyces marxiannus, and tested against ?-amylases from Callosobruchus maculates and human saliva. The D2 fraction presented higher antimicrobial activity and was subjected to further purification and seven new different fractions (H1-H7) were obtained. Peptides in the H4 fraction were sequenced and the N-terminal sequences revealed homology with previously reported storage vicilins from seeds. The H4 fraction exhibited strong antifungal activity and also promoted morphological changes in yeast, including pseudohyphae formation. All fractions, including H4, inhibited mammalian ?-amylase activity but only the H4 fraction was able to inhibit C. maculatus ?-amylase activity. These results suggest that the fractions isolated from the seeds of C. baccatum can act directly in plant defenses against pathogens and insects. © 2014 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 102: 335-343, 2014. PMID:24817604

Vieira Bard, Gabriela C; Nascimento, Viviane V; Oliveira, Antônia Elenir A; Rodrigues, Rosana; Da Cunha, Maura; Dias, Germana B; Vasconcelos, Ilka M; Carvalho, Andre O; Gomes, Valdirene M

2014-04-01

250

Identification and analysis of the amylase-binding protein B (AbpB) and gene ( abpB) from Streptococcus gordonii  

Microsoft Academic Search

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

Lina Li; Jason M. Tanzer; Frank A. Scannapieco

2002-01-01

251

High pressure homogenization of a fungi ?-amylase  

Microsoft Academic Search

The activity and stability of ?-amylase after high pressure homogenization were investigated. The enzyme buffer solution was processed at homogenization pressures up to 1500bar. No changes in the enzymatic activity at 15, 45 and 75°C were observed after the homogenization process. The evaluation of calcium requirement to preserve the ?-amylase stability during homogenization was carried out and the results indicated

Alline Artigiani Lima Tribst; Marcelo Cristianini

252

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

PubMed Central

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

2012-01-01

253

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

PubMed

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

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

2014-01-01

254

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

PubMed Central

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

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

2014-01-01

255

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

PubMed Central

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

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

2011-01-01

256

Amylase creatinine clearance ratio following biliary tract surgery  

Microsoft Academic Search

An increase in the amylase creatinine clearance ratio (ACCR) is considered to be a more specific diagnostic test of acute pancreatitis than elevation of the serum or urine amylase activity. Simultaneous serum and urine samples for the determination of amylase and creatinine were obtained preoperatively and at various intervals postoperatively in 75 patients undergoing surgery. No increase in ACCR was

Francesco Tonelli; Emidio Senati; Marileda Indinnimeo

1981-01-01

257

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

Microsoft Academic Search

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

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

2009-01-01

258

Salivary gland acinar cells regenerate functional glandular structures in modified hydrogels  

NASA Astrophysics Data System (ADS)

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

Pradhan, Swati

259

Human salivary MUC7 mucin peptides: effect of size, charge and cysteine residues on antifungal activity.  

PubMed Central

We have previously shown that MUC7 (human salivary low-molecular-mass mucin) 20-mer: LAHQKPFIRKSYKCLHKRCR (residues 32-51 of the parent MUC7, with a net positive charge of 7) possesses a broad-spectrum antimicrobial activity [Bobek and Situ (2003) Antimicrob. Agents Chemother. 47, 645-652]. The aims of the present study were to determine the minimum peptide chain length and its location within the 20-mer region that retains potent antifungal activity against Candida albicans and Cryptococcus neoformans and to examine the effect of net charge of the peptide as well as the role of cysteine residues on the fungicidal activity. First, several C-terminal truncated MUC7 20-mer fragments (16-mer, 12-mer, 11-mer, 10-mer and 8-mer) and one N-terminal fragment (8-mer-N) were synthesized and tested. The results showed that MUC7 12-mer, located at the C-terminal region of MUC7 20-mer, having a net charge of +6 and exhibiting an amphipathic helical conformation, not only retained but exceeded the antifungal activity of that of 20-mer. Secondly, several variants of the 12-mer peptide containing a lower or no net positive charge, or no cysteine residues were synthesized and tested. A clear correlation between the net positive charge of the 12-mer, its potency and initial interaction of peptide with fungal cells was found by killing assays, fluorescence microscopy and fungal cell-membrane potential measurements. Furthermore, cysteine residues, which play a critical role in bacterial binding, were found to be not important for the fungicidal activity of these peptides. These results identified MUC7 12-mer as a potential candidate for development into a novel antifungal therapeutic agent.

Situ, Hongsa; Wei, Guoxian; Smith, Christina J; Mashhoon, Shirin; Bobek, Libuse A

2003-01-01

260

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

PubMed Central

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

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

2012-01-01

261

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

Microsoft Academic Search

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

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

2011-01-01

262

Salivary Gland Cancer  

MedlinePLUS

... a roadmap to this full guide. About the salivary glands The salivary glands are tissues that produce saliva. ... are often called the minor salivary glands. About salivary gland cancer Cancer begins when normal cells change and ...

263

Apyrase activity and adenosine diphosphate induced platelet aggregation inhibition by the salivary gland proteins of Culicoides variipennis, the North American vector of bluetongue viruses.  

PubMed

Salivary gland homogenates of Culicoides variipennis, the primary vector of bluetongue (BLU) viruses in North America, were analyzed for apyrase activity. Apyrase (ATP diphosphohydrolase, EC 3.6.1.5) is an anti-hemostatic and anti-inflammatory salivary enzyme of most hematophagous arthropods. The enzyme activity was measured by the release of orthophosphate using ATP, ADP, and AMP as substrates with Ca2+ as the divalent cation. ATPase (11.5 +/- 1 mU/pair of glands), ADPase (7.3 +/- 0.7 mU/pair of glands), and insignificant (P < 0.05) AMPase (0.07 mU/pair of glands) activities were detected in female salivary glands. Male salivary glands contained lower amounts of ATPase and ADPase activity (P < 0.05). The ATPase and ADPase activities were greatest at pH 8.5, and were similarly activated by Mg2+. Molecular sieving HPLC of salivary gland homogenates generated a single peak which coincided with ATPase and ADPase, but no AMPase, activity; the protein has an estimated molecular mass of 35,000 Da. ATPase and ADPase activity, and total protein concentration, were reduced (P < 0.05) in the salivary glands of females after taking a blood meal from a sheep. Salivary gland homogenates also inhibited ADP-induced platelet aggregation in vitro. It is concluded that the salivary ATPase and ADPase activities of C. variipennis reside in one enzyme, and that this enzyme is likely an apyrase. The apyrase activity is thought to be responsible for the inhibition of ADP-induced platelet aggregation, as indicated by the apparent discharge of apyrase from salivary glands into the host during blood feeding. This suggests that apyrase is one of the salivary proteins present in C. variipennis acting as antigens in the development of Culicoides hypersensitivity in ruminants and horses. Apyrase may inhibit an inflammatory response at the feeding site through the subsequent degradation of its end-product, AMP, to adenosine, a potent anti-inflammatory substance, by the ecto-5' nucleotidase activity of neutrophils. PMID:8720570

Pérez de León, A A; Tabachnick, W J

1996-02-01

264

Adherence of oral streptococci to salivary glycoproteins.  

PubMed Central

We used an overlay method to study the ability of human salivary glycoproteins to serve as receptors for several strains of streptococci that colonize the oral cavity. Parotid and submandibular-sublingual salivas were collected as ductal secretions, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and transferred to nitrocellulose membranes. The resulting blots were overlaid with [35S]methionine-labeled bacteria, and salivary components to which the bacteria bound were detected by autoradiography. Potential glycoprotein receptors were identified for 8 of the 16 strains tested. In three cases (Streptococcus sanguis 72-40 and 804 and Streptococcus sobrinus OMZ176), highly specific interactions with a single salivary component were detected. Removal of sialic acid residues from the low-molecular-weight salivary mucin prevented adherence of one of these strains (S. sanguis 72-40), suggesting that this saccharide either mediates binding or is a critical component of the receptor site. In the remaining five strains (Streptococcus gordonii G9B and 10558, S. sanguis 10556, and Streptococcus oralis 10557 and 72-41), interactions with multiple salivary components, including the low-molecular-weight salivary mucin, highly glycosylated proline-rich glycoproteins, and alpha-amylase, were detected. These results suggest that some oral streptococci can bind specifically to certain of the salivary glycoproteins. The interactions identified may play an important role in governing bacterial adherence and clearance within the oral cavity. Images

Murray, P A; Prakobphol, A; Lee, T; Hoover, C I; Fisher, S J

1992-01-01

265

Rapid amylase and lipase determinations by nephelometry.  

PubMed

The Coleman 91 nephelometer provides rapid, simple amylase and lipase assays, which are particularly suited to emergency requests. Linearity of amylase compares favorably with that of the Phadebas assay, and comparable precision was obtainable with serum and urine. Normal ranges for serum amylase are slightly higher than those based on the amyloclastic end-point assay. The serum lipase assay shows improved linearity over titrimetric procedures, although kinetics remain variably non-linear. Occasional sera show discordantly elevated nephelometric lipase and normal titrimetric lipase. Precision of the nephelometric lipase assay is somewhat lower than that of amylase; normal ranges are considerably higher than those based on titrimetry. Extremely lactescent sera may yield falsely low nephelometric amylase and lipase activities: these sera must be serially diluted to achieve actual values. PMID:696672

Yourno, J; Henry, J B

1978-07-01

266

A heterogeneous beta-D-glucosidase activator from monkey parotid gland and its use as an affinity ligand in the purification of human salivary beta-D-glucosidase.  

PubMed

We have isolated a heat-stable, low molecular weight activator peptide(s) from monkey parotid gland that specifically activated human salivary beta-D-glucosidase. This activator appeared to be heterogeneous on Sephadex G-25 gel filtration and polyacrylamide gel electrophoresis under non-denaturing conditions. About 45% of the human salivary beta-glucosidase could bind to the activator immobilised on Sepharose and be eluted by Cutscum. The purified enzyme was nearly homogeneous, with a subunit Mr of 46,000 as revealed by SDS-gel electrophoresis and silver staining. PMID:3107568

Nagarajan, S; Cherian, R; Balasubramanian, A S

1987-02-01

267

The value of alpha-amylase and isoamylase determination in chronic renal failure patients.  

PubMed

Hyperamylasemia is a common finding in chronic renal failure (CRF) patients. It has been suggested that the diagnosis of acute pancreatitis in these patients is confirmed when serum amylase activities are greater than three times the upper normal limits. In order to evaluate the frequency, the type, and the hyperamylasemia levels in patients with various degree of chronic renal failure, the total serum amylase (Ta), the pancreatic (Pa) and salivary (Sa) types of serum isoamylases, as well as the urine isoamylases (Tu, Pu, Su, respectively) have been determined by the Phadebas method. Moreover, the levels of serum electrolytes and triglycerides were determined in order to study any relationship between serum electrolytes as well as triglycerides and alpha-amylase activities. We studied 102 patients of whom 33 (group A) had CRF with serum creatinine levels 8.5 +/- 3.1 mg/dL (mean +/- SD), 59 (group B) were receiving chronic hemodialysis, and 10 (group C) were on continuous ambulatory peritoneal dialysis as well as 47 normal individuals. None of the subjects studied had any clinical manifestation of acute pancreatitis. Our results showed that the Ta, Pa and Sa levels of groups A, B, and C were significantly elevated compared to normal subjects. Eighteen patients had Pa activities greater than three times the upper normal limits. In the present study, no relationship between serum electrolytes as well as triglycerides and alpha-amylases was found. In conclusion, hyperamylasemia was a much more common finding in CRF patients than previously reported in the literature.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8071568

Tsianos, E V; Dardamanis, M A; Elisaf, M; Vasakos, S; Siamopoulos, K C

1994-04-01

268

Active sites of salivary proline-rich protein for binding to Porphyromonas gingivalis fimbriae.  

PubMed Central

Porphyromonas gingivalis fimbriae specifically bind salivary acidic proline-rich protein 1 (PRP1) through protein-protein interactions. The binding domains of fimbrillin (a subunit of fimbriae) for PRP1 were analyzed previously (A. Amano, A. Sharma, J.-Y. Lee, H. T. Sojar, P. A. Raj, and R. J. Genco, Infect. Immun. 64:1631-1637, 1996). In this study, we investigated the sites of binding of the PRP1 molecules to the fimbriae. PRP1 (amino acid residues 1 to 150) was proteolysed to three fragments (residues 1 to 74 [fragment 1-74], 75 to 129, and 130 to 150). 125I-labeled fimbriae clearly bound fragments 75-129 and 130-150, immobilized on a polyvinylidene difluoride membrane; both fragments also inhibited whole-cell binding to PRP1-coated hydroxyapatite (HAP) beads by 50 and 83%, respectively. However, the N-terminal fragment failed to show any effect. Analogous peptides corresponding to residues 75 to 89, 90 to 106, 107 to 120, 121 to 129, and 130 to 150 of PRP1 were synthesized. The fimbriae significantly bound peptide 130-150, immobilized on 96-well plates, and the peptide also inhibited binding of 125I-labeled fimbriae to PRP1-coated HAP beads by almost 100%. Peptides 75-89, 90-106, and 121-129, immobilized on plates, showed considerable ability to bind fimbriae. For further analysis of active sites in residues 130 to 150, synthetic peptides corresponding to residues 130 to 137, 138 to 145, and 146 to 150 were prepared. Peptide 138-145 (GRPQGPPQ) inhibited fimbrial binding to PRP1-coated HAP beads by 97%. This amino acid sequence was shared in the alignment of residues 75 to 89, 90 to 106, and 107 to 120. Six synthetic peptides were prepared by serial deletions of individual residues from the N and C termini of peptide GRPQGPPQ. Peptide PQGPPQ was as inhibitory as peptide GRPQGPPQ. Further deletions of the dipeptide Pro-Gln from the N and C termini of peptide PQGPPQ resulted in significant loss of the inhibitory effect. These results strongly suggest that PQGPPQ is the minimal active segment for binding to P. gingivalis fimbriae and that the moiety of the Pro-Gln dipeptide plays a critical role in expressing binding ability.

Kataoka, K; Amano, A; Kuboniwa, M; Horie, H; Nagata, H; Shizukuishi, S

1997-01-01

269

Active sites of salivary proline-rich protein for binding to Porphyromonas gingivalis fimbriae.  

PubMed

Porphyromonas gingivalis fimbriae specifically bind salivary acidic proline-rich protein 1 (PRP1) through protein-protein interactions. The binding domains of fimbrillin (a subunit of fimbriae) for PRP1 were analyzed previously (A. Amano, A. Sharma, J.-Y. Lee, H. T. Sojar, P. A. Raj, and R. J. Genco, Infect. Immun. 64:1631-1637, 1996). In this study, we investigated the sites of binding of the PRP1 molecules to the fimbriae. PRP1 (amino acid residues 1 to 150) was proteolysed to three fragments (residues 1 to 74 [fragment 1-74], 75 to 129, and 130 to 150). 125I-labeled fimbriae clearly bound fragments 75-129 and 130-150, immobilized on a polyvinylidene difluoride membrane; both fragments also inhibited whole-cell binding to PRP1-coated hydroxyapatite (HAP) beads by 50 and 83%, respectively. However, the N-terminal fragment failed to show any effect. Analogous peptides corresponding to residues 75 to 89, 90 to 106, 107 to 120, 121 to 129, and 130 to 150 of PRP1 were synthesized. The fimbriae significantly bound peptide 130-150, immobilized on 96-well plates, and the peptide also inhibited binding of 125I-labeled fimbriae to PRP1-coated HAP beads by almost 100%. Peptides 75-89, 90-106, and 121-129, immobilized on plates, showed considerable ability to bind fimbriae. For further analysis of active sites in residues 130 to 150, synthetic peptides corresponding to residues 130 to 137, 138 to 145, and 146 to 150 were prepared. Peptide 138-145 (GRPQGPPQ) inhibited fimbrial binding to PRP1-coated HAP beads by 97%. This amino acid sequence was shared in the alignment of residues 75 to 89, 90 to 106, and 107 to 120. Six synthetic peptides were prepared by serial deletions of individual residues from the N and C termini of peptide GRPQGPPQ. Peptide PQGPPQ was as inhibitory as peptide GRPQGPPQ. Further deletions of the dipeptide Pro-Gln from the N and C termini of peptide PQGPPQ resulted in significant loss of the inhibitory effect. These results strongly suggest that PQGPPQ is the minimal active segment for binding to P. gingivalis fimbriae and that the moiety of the Pro-Gln dipeptide plays a critical role in expressing binding ability. PMID:9234769

Kataoka, K; Amano, A; Kuboniwa, M; Horie, H; Nagata, H; Shizukuishi, S

1997-08-01

270

Salivary duct stones  

MedlinePLUS

... of minerals in the ducts that drain the salivary glands. Salivary duct stones are a type of salivary gland disorder. ... Saliva (spit) is produced by the salivary glands in the mouth. The ... that can block the salivary ducts. When saliva cannot exit ...

271

The effect of starvation on the diurnal variation of amylase secretion from rat parotid glands  

Microsoft Academic Search

The purpose of this study was to investigate the effect of starvation on the cellular function of rat parotid glands in relation\\u000a to the diurnal variation of amylase secretion from the tissue. Salivary amylase secreted from the glands of rats starved for\\u000a 24h showed diurnal variation, with two peaks at 13 and 21h. The peak at 21 h was more

Yasuko Ishikawa; Cang Chen; Hajime Ishida

1993-01-01

272

Prevalence of the Amylase-Binding Protein A Gene (abpA )i n Oral Streptococci  

Microsoft Academic Search

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 Strepto- coccus 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.

ALAN E. BROWN; JEFFREY D. ROGERS; ELAINE M. HAASE; PETER M. ZELASKO

1999-01-01

273

Purification of ?-amylase from Bacillus lentus cultures  

Microsoft Academic Search

Acetone fractionation of Bacillus lentus culture filtrate yielded the highest a-amylase activity and the 66.6% fraction reached 13-fold that of the crude enzyme preparation. Gel filtration and ion exchange chromatography afforded a pure a-amylase (relative molecular mass, 42 000). The pure enzyme was highly active on starch and dextrin. It produced a mixture of oligosaccharides as major products of starch

S. A. El-Aassar; S. H. Omar; M. K. Gouda; A. M. Ismail; A. F. Abdel-Fattah

1992-01-01

274

What Is Salivary Gland Cancer?  

MedlinePLUS

... key statistics about salivary gland cancer? What is salivary gland cancer? Salivary gland cancer starts in one of ... tumors can develop in these glands. About the salivary glands Salivary glands make saliva – the lubricating fluid found ...

275

Salivary gland tumors  

MedlinePLUS

Salivary gland tumors are abnormal cells growing in the ducts that drain the salivary glands. ... The salivary glands are located around the mouth. They produce saliva, which moistens food to help with chewing and swallowing. ...

276

Radionuclide salivary gland imaging  

SciTech Connect

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

Mishkin, F.S.

1981-10-01

277

Salivary pellicles.  

PubMed

The salivary pellicle is a thin acellular organic film that forms on any type of surface upon exposure to saliva. The role of the pellicle is manifold, and it plays an important role in the maintenance of oral health. Its functions include not only substratum protection and lubrication, but also remineralization and hydration. It also functions as a diffusion barrier and possesses buffering ability. Not only the function, but also the formation, composition and stability of the pellicle are known to be highly influenced by the physicochemical properties of both substrata and ambient media. In this chapter, we discuss these aspects of salivary pellicles, an area where research has boomed in the past years partly because of the application of experimental techniques often reserved for more traditional surface science studies. © 2014 S. Karger AG, Basel. PMID:24862592

Lindh, Liselott; Aroonsang, Watcharapong; Sotres, Javier; Arnebrant, Thomas

2014-01-01

278

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

PubMed Central

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

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

2010-01-01

279

Activation of Salivary Secretion: Coupling of Cell Volume and [Ca2+]i in Single Cells  

NASA Astrophysics Data System (ADS)

High-resolution differential interference contrast microscopy and digital imaging of the fluorescent calcium indicator dye fura-2 were performed simultaneously in single rat salivary gland acinar cells to examine the effects of muscarinic stimulation on cell volume and cytoplasmic calcium concentration ([Ca2+]i). Agonist stimulation of fluid secretion is initially associated with a rapid tenfold increase in [Ca2+]i as well as a substantial cell shrinkage. Subsequent changes of cell volume in the continued presence of agonist are tightly coupled to dynamic levels of [Ca2+]i, even during [Ca2+]i oscillations. Experiments with Ca2+ chelators and ionophores showed that physiological elevations of [Ca2+]i are necessary and sufficient to cause changes in cell volume. The relation between [Ca2+]i and cell volume suggests that the latter reflects the secretory state of the acinar cell. Agonist-induced changes in [Ca2+]i, by modulating specific ion permeabilities, result in solute movement into or out of the cell. The resultant cell volume changes may be important in modulating salivary secretion.

Foskett, J. Kevin; Melvin, James E.

1989-06-01

280

Culture Conditions for Production of Thermostable Amylase by Bacillus stearothermophilus  

PubMed Central

Bacillus stearothermophilus grew better on complex and semisynthetic medium than on synthetic medium supplemented with amino acids. Amylase production on the complex medium containing beef extract or corn steep liquor was higher than on semisynthetic medium containing peptone (0.4%). The synthetic medium, however, did not provide a good yield of extracellular amylase. Among the carbohydrates which favored the production of amylase are, in order starch > dextrin > glycogen > cellobiose > maltohexaose-maltopeptaose > maltotetraose and maltotriose. The monosaccharides repressed the enzyme production, whereas inositol and d-sorbitol favored amylase production. Organic and inorganic salts increased amylase production in the order of KCI > sodium malate > potassium succinate, while the yield was comparatively lower with other organic salts of Na and K. Amino acids, in particular isoleucine, cysteine, phenylalanine, and aspartic acids, were found to be vital for amylase synthesis. Medium containing CaCl2 2H2O enhanced amylase production over that on Ca2+ -deficient medium. The detergents Tween-80 and Triton X-100 increased biomass but significantly suppressed amylase synthesis. The amylase powder obtained from the culture filtrate by prechilled acetone treatment was stable over a wide pH range and liquefied thick starch slurries at 80°C. The crude amylase, after (NH4)2SO4 fractionation, had an activity of 210.6 U mg?1. The optimum temperature and pH of the enzyme were found to be 82°C and 6.9, respectively. Ca2+ was required for the thermostability of the enzyme preparation.

Srivastava, R. A. K.; Baruah, J. N.

1986-01-01

281

Immobilization of mold and bacterial amylases on silica carriers.  

PubMed

The immobilization of alpha-amylase and glucoamylase was investigated by several coupling methods on silica carriers, different types of Silokhroms, and silica gels. The most active immobilized mold and bacterial alpha-amylases and mold glucoamylase were obtained with titanium salts. These activities were twice the value of that obtained by glutaraldehyde or azo coupling. The half-lives of A. oryzae alpha-amylase, B. subtilis alpha-amylase, and A. niger glucoamylase, immobilized to silica carriers at 45 degrees C and under continuous operation at a high concentration of substrate, were 14, 35, and 65 days, respectively. PMID:18548434

Kvesitadze, G I; Dvali, M S

1982-08-01

282

Salivary gland dysfunction following radioactive iodine therapy  

SciTech Connect

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.

Wiesenfeld, D.; Webster, G.; Cameron, F.; Ferguson, M.M.; MacFadyen, E.E.; MacFarlane, T.W.

1983-02-01

283

Hypoglycemic effect of basil (Ocimum basilicum) aqueous extract is mediated through inhibition of ?-glucosidase and ?-amylase activities: an in vitro study.  

PubMed

The present study investigated the in vitro hypoglycemic activity of basil (Ocimum basilicum) aqueous extract. Preliminary phytochemical screening of the extract revealed the presence of reducing sugars, cardiac glycosides, tannins, saponins, glycosides, flavonoids and steroids. The total polyphenols content (TPC), flavonoids content (FC), percentage diphenylpicrylhydrazyl (DPPH( · )) radical inhibition and total antioxidant status (TAS) were estimated. The FC was 41 ± 2.2 rutin/g dry extract, the TPC was 146 ± 5.26 mg catechin/g dry extract and the TAS was 5.12 ± 0.7 mmol/L. The %DPPH( · ) free radical inhibition was 60%, 54%, 49% and 43%, respectively, for different extract concentrations; 20, 18.2, 16.3 and 14.5 mg/ml, respectively. The extract elicited significant dose-dependent pattern against rat intestinal sucrase (RIS; IC(50) = 36.72 mg/ml), rat intestinal maltase (RIM; IC(50) = 21.31 mg/ml) and porcine pancreatic ?-amylase (PPA; IC(50) = 42.50 mg/ml) inhibitory activities. The inhibition was greater against maltase compared with sucrase. These effects may be attributed to the high TPC and FC levels. The linear regression analysis revealed strong significant positive correlations between %DPPH( · ) radical inhibition and each of %RIS, %RIM and %PPA inhibiting activity. Also, strong significant positive correlations between %RIS and either %RIM or %PPA inhibition activity were observed. We concluded therefore that basil aqueous extract via antioxidant and possibly ?-glucosidase and ?-amylase inhibiting activities, offered positive benefits to control diabetes. PMID:21636683

El-Beshbishy, Ha; Bahashwan, Sa

2012-02-01

284

Production of amylase by Arthrobacter psychrolactophilus.  

PubMed

Arthrobacter psychrolactophilus ATCC 700733 grew with a doubling time of 1.5-2.3 h (22 degrees C) and produced up to 0.2 units/mL (soluble starch assay) of extracellular amylase in tryptic soy broth without dextrose (TSBWD) containing 0.5% or 1.0% (w/v) soluble starch or maltose as the fermentable substrate. Time-course experiments in media containing soluble starch as substrate showed that amylolytic activity appeared in cultures at 24 h (after exponential growth had ceased), reached peak levels in 72-96 h, and declined rapidly after reaching peak levels. Peak levels were highest in TSBWD containing 1.0% soluble starch. Proteolytic activity appeared at about the same time as amylolytic activity and increased during the period of amylase production. Significant amylase production was not observed in cultures in TSBWD with 0.5% glucose or in cultures grown at 28 degrees C, but low levels of amylase were observed in TSBWD cultures grown at 19-23 degrees C which contained no added carbohydrate. A single band of activity was observed after electrophoresis of supernatant fractions in non-denaturing gels, followed by in situ staining for amylolytic activity. The amylase possessed a raw starch-binding domain and bound to uncooked corn, wheat or potato starch granules. It was active in the Phadebas assay for alpha-amylase. Activity was maximum on soluble starch at a temperature between 40 degrees C and 50 degrees C. The amylase after purification by affinity chromatography on raw starch granules exhibited two starch-binding protein bands on SDS gels of 105 kDa and 26 kDa. PMID:15931519

Smith, Michael R; Zahnley, James C

2005-07-01

285

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

PubMed

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

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

2014-03-01

286

Mapping of sequence-specific markers and loci controlling preharvest sprouting and alpha-amylase activity in rye (Secale cereale L.) on the genetic map of an F2 (S120×S76) population.  

PubMed

Location of the loci that control preharvest sprouting and alpha-amylase activity in rye was studied based on intercross S120×S76, consisting of 110 genotypes of F2 and F3 progenies. The genetic map currently consists of 141 loci distributed in 11 linkage groups, covering a distance of 506.4 cM, and was enriched during this study with 24 sequence-specific markers (7 SCARs, 7 SSRs, and 10 STSs). The extended map was applied for composite interval mapping of the loci controlling preharvest sprouting and ?-amylase activity, revealing 3 significant QTLs for preharvest sprouting, located on chromosomes 3R, 5R and 6R (in 1999), and one QTL for ?-amylase activity found on chromosome 2R (in 2000). PMID:20720302

Myskow, B; Stojalowski, S; Milczarski, P; Masojc, P

2010-01-01

287

Electrophoretic separation, detection, and variation of amylase isoenzymes.  

PubMed

An electrophoretic technique for demonstrating amylase isoenzymes is described. After separation in an agarose gel containing a linear polyacrylamide polymer to reduce electroendosmotic flow, the amylase fractions are visualized by incubation with a commercially available dye-starch polymer (Phadebas Amylase Test). The technique detects amylase fractions with activities below 10 U/l. Some characteristic changes in such diseases as acute and chronic pancreatitis, cystic fibrosis of the pancreas, macroamylasemia and inherited variants as well as after maxillofacial surgery are mentioned. PMID:48275

Skude, G

1975-01-01

288

Detection of the quantitative trait loci for ?-amylase activity on a high-density genetic map of rye and comparison of their localization to loci controlling preharvest sprouting and earliness.  

PubMed

The objectives of the research were to determine the position of quantitative trait loci (QTL) for ?-amylase activity on the genetic map of a rye recombinant inbred line population-S120 × S76-and to compare them to known QTL for preharvest sprouting and heading earliness. Fourteen QTL for ?-amylase activity on all seven chromosomes were identified. The detected QTL were responsible for 6.09-23.32% of ?-amylase activity variation. The lowest LOD value (2.22) was achieved by locus QAa4R-M3 and the highest (7.79) by locus QAa7R-M1. Some QTL intervals for features of interest overlapped partially or completely. There were six overlapping QTL for ?-amylase activity and preharvest sprouting (on 1R, 3R, 4R, 6R, 7R) and the same number for preharvest sprouting and heading earliness (on 1R, 2R, 6R, 7R). Furthermore, there was one interval partially common to all three traits, mapped on the long arm of chromosome 1R. Testing of lines originating from hybrid breeding programs, such as S120 and S76, may provide important information about the most significant genes and markers for selection in commercial breeding. Among the statistically significant markers selected in the Kruskal-Wallis test (P < 0.005), there were 55 common ones for preharvest sprouting and heading earliness (1R, 2R, 6R), 30 markers coinciding between ?-amylase activity and preharvest sprouting (5R, 7R) and one marker for ?-amylase activity and heading earliness (6R). ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11032-011-9627-1) contains supplementary material, which is available to authorized users. PMID:22707913

My?ków, Beata; Stoja?owski, Stefan; La?, Anna; Bolibok-Br?goszewska, Hanna; Rakoczy-Trojanowska, Monika; Kilian, Andrzej

2012-06-01

289

A critical assessment of a viscometric assay for measuring Saccharomycopsis fibuligera ?-amylase activity on gelatinised cassava starch  

Microsoft Academic Search

A viscometric technique for measuring Saccharomycopsis fibuligera DSM-70554 ?-amylase on gelatinised cassava starch aqueous solutions was assessed. The selected conditions for working over a reliable viscosity measurement range involved a starch concentration of 5% (w\\/v) and a shear rate of 0.168 1\\/s. Viscometric assay involved the determination of the slope of the decrease in viscosity with time of the starch

C. F González; J. I Farińa; L. I. C Figueroa

2002-01-01

290

Study of growth factors, alpha-amylase and peroxidase activity in various cultivars of rice (Oryza sativa L.) under vanillic acid stress.  

PubMed

The effects of Vanillic Acid (VA) on germination, seedling and adult plant of rice (Oryza sativa L.) were investigated. Four cultivars, traditional (Taroom mahalli and Taroom deilamani) and improved (Shafagh and Onda) were studied. For germination, seeds were sterilized and then placed on Petri dish at 30 degrees C at different concentrations (0, 10, 20 and 25 mM) for 7 days and growth factors of seedling were measured after 14 days. Seedling (10 days) planted in hydroponic medium including nutrient solution amended with 0, 25, 50 and 100 mg kg(-1) VA. After 30 days, the growth factors were determined, alpha-amylase and peroxidase activities were assayed by Bernfeld and Maehly methods, respectively. Present results indicated that as a result of increasing concentrations of VA, germination percentage and germination rate as well as alpha-amylase activity markedly decreased except Onda in all cultivars. In all cultivars, seedling growth factors such as shoot length, fresh weight and dry weight as well as root have been reduced by increasing of VA. In adult plant, also shoot length, root length, shoot dry weight and root fresh weight were reduced by increasing concentrations of VA. Shoot fresh weight was decreased in Shafagh by increasing concentrations of VA; meanwhile, we have not observed any significant differences in the other cultivars. In the case of root dry weight, there were not significant differences in any cultivars. With increasing concentrations of VA, chlorophyll content reduced; on the contrary, peroxidase activity increased. PMID:19086516

Jazayeri, O; Aghajanzadeh, T A; Gildeh, B Sadeghpour

2007-05-15

291

DE NOVO SYNTHESIS OF ?-AMYLASE BY BACILLUS STEAROTHERMOPHILUS1  

PubMed Central

Welker, N. E. (Western Reserve University, Cleveland, Ohio and University of Illinois, Urbana), and L. Leon Campbell. De novo synthesis of ?-amylase by Bacillus stearothermophilus. J. Bacteriol. 86:1202–1210. 1963.—The pH optimum for the synthesis of ?-amylase by washed-cell suspensions was 6.7. ?-Amylase synthesis began soon after the addition of the inducer (maltose, methyl-?-d-maltoside, or phenyl-?-d-glucoside, at 10?3m), proceeded at a linear rate for 60 min, and then leveled off. Cell suspensions without inducer produced small amounts of ?-amylase. The addition of glucose (2 × 10?3m), sucrose (10?3m), or glycerol (4 × 10?3m) to washed-cell suspensions failed to stimulate the production of ?-amylase. Nitrogen starvation of washed cells for 60 min with fructose as a carbon source or by induction with pure maltose showed that the ability to produce ?-amylase was lost. Examination of the amino acid pool at this time showed a general depletion of amino acids and the complete disappearance of tyrosine, phenyl-alanine, proline, and valine. Replenishment of the amino acid pool with casein hydrolysate (0.5%) restored the ability of the cells to produce ?-amylase. Chloramphenicol and 8-azaguanine were shown to inhibit ?-amylase synthesis. Inhibition was observed immediately upon the addition of chloramphenicol to cell suspensions preinduced for varying periods of time. Actinomycin D and mitomycin C also inhibited ?-amylase synthesis when added to induced washed-cell suspensions. The amino acid analogues, norvaline, norleucine, and ethionine, inhibited ?-amylase formation by 72, 53, and 38%, respectively. p-Fluorophenylalanine inhibited the synthesis of active ?-amylase by 92% and the incorporation of proline-C14 into ?-amylase and cellular proteins by 95 and 74%, respectively.

Welker, N. E.; Campbell, L. Leon

1963-01-01

292

DE NOVO SYNTHESIS OF ALPHA-AMYLASE BY BACILLUS STEAROTHERMOPHILUS.  

PubMed

Welker, N. E. (Western Reserve University, Cleveland, Ohio and University of Illinois, Urbana), and L. Leon Campbell. De novo synthesis of alpha-amylase by Bacillus stearothermophilus. J. Bacteriol. 86:1202-1210. 1963.-The pH optimum for the synthesis of alpha-amylase by washed-cell suspensions was 6.7. alpha-Amylase synthesis began soon after the addition of the inducer (maltose, methyl-beta-d-maltoside, or phenyl-alpha-d-glucoside, at 10(-3)m), proceeded at a linear rate for 60 min, and then leveled off. Cell suspensions without inducer produced small amounts of alpha-amylase. The addition of glucose (2 x 10(-3)m), sucrose (10(-3)m), or glycerol (4 x 10(-3)m) to washed-cell suspensions failed to stimulate the production of alpha-amylase. Nitrogen starvation of washed cells for 60 min with fructose as a carbon source or by induction with pure maltose showed that the ability to produce alpha-amylase was lost. Examination of the amino acid pool at this time showed a general depletion of amino acids and the complete disappearance of tyrosine, phenyl-alanine, proline, and valine. Replenishment of the amino acid pool with casein hydrolysate (0.5%) restored the ability of the cells to produce alpha-amylase. Chloramphenicol and 8-azaguanine were shown to inhibit alpha-amylase synthesis. Inhibition was observed immediately upon the addition of chloramphenicol to cell suspensions preinduced for varying periods of time. Actinomycin D and mitomycin C also inhibited alpha-amylase synthesis when added to induced washed-cell suspensions. The amino acid analogues, norvaline, norleucine, and ethionine, inhibited alpha-amylase formation by 72, 53, and 38%, respectively. p-Fluorophenylalanine inhibited the synthesis of active alpha-amylase by 92% and the incorporation of proline-C(14) into alpha-amylase and cellular proteins by 95 and 74%, respectively. PMID:14086090

WELKER, N E; CAMPBELL, L L

1963-12-01

293

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

PubMed

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

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

2004-03-01

294

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

PubMed Central

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

Singh, Sanamdeep; Bali, Vrinda; Mangla, Jyoti

2014-01-01

295

Salivary contribution to exhaled nitric oxide  

Microsoft Academic Search

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

W. Zetterquist; C. Pedroletti; J. O. n. Lundberg; K. Alving

1999-01-01

296

Identification and analysis of a gene (abpA) encoding a major amylase-binding protein in Streptococcus gordonii  

Microsoft Academic Search

Oral streptococci such as Streptococcus gordonii bind the abundant salivary enzyme a-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

Jeffrey D. Rogers; Elaine M. Haase; Alan E. Brown; Charles W. I. Douglas; Justin P. Gwynn; Frank A. Scannapieco

1998-01-01

297

Study of the Saccharogenic Method for the Determination of Serum and Urine Amylase  

Microsoft Academic Search

ECAUSE OF THE INDUSTRIAL importance of amylases, the literature on the actions of amyiases on starch is extensive, going back into the nineteenth century (1, 2). Only a few of these investigations have been concerned specifically with the cz-amylase activity of serum and urine. Methods for the determination of amylase in body fluids have included viscosimetric, turbidimetric, iodometric (amyloclastic), and

Richard J. Henry; Neil Chiamori

298

Late-maturity ?-amylase: Low falling number in wheat in the absence of preharvest sprouting  

Microsoft Academic Search

Late maturity ?-amylase (LMA), or prematurity ?-amylase (PMAA) as it has been termed in the UK, in wheat involves the untimely synthesis of high pI ?-amylase during the middle to later stages of grain development and ripening. The enzyme activity is retained in the grain at harvest ripeness, resulting in low falling number and failure to meet receival standards and

Daryl Mares; Kolumbina Mrva

2008-01-01

299

The effectiveness of the Uchida-Kraepelin test for psychological stress: an analysis of plasma and salivary stress substances  

PubMed Central

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.

Sugimoto, Koreaki; Kanai, Aya; Shoji, Noriaki

2009-01-01

300

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

PubMed

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

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

2013-03-01

301

Immobilization of ?-Amylase onto Luffa operculata Fibers  

PubMed Central

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

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

2013-01-01

302

Flaxseed (Linum usitatissimum L.) extract as well as (+)-secoisolariciresinol diglucoside and its mammalian derivatives are potent inhibitors of ?-amylase activity.  

PubMed

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 3.2.1.1) by flaxseed-derived lignans that could therefore constitute a promising nutraceutical for the prevention and the treatment of T2DM. PMID:23583514

Hano, Christophe; Renouard, Sullivan; Molinié, Roland; Corbin, Cyrielle; Barakzoy, Esmatullah; Doussot, Joël; Lamblin, Frédéric; Lainé, Eric

2013-05-15

303

Salivary gland infections  

MedlinePLUS

Salivary gland infections affect the glands that produce saliva (spit). The infection may be due to bacteria or viruses. There are three pairs of major salivary glands: Parotid glands. These are the two largest glands. ...

304

Salivary Gland Disorders  

MedlinePLUS

Your salivary glands make saliva - sometimes called spit - and empty it into your mouth through openings called ducts. Saliva makes ... contains antibodies that can kill germs. Problems with salivary glands can cause the glands to become irritated and ...

305

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

PubMed

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

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

2011-02-01

306

CE-LIF determination of salivary cadaverine and lysine concentration ratio as an indicator of lysine decarboxylase enzyme activity.  

PubMed

Salivary bacteria produce the enzyme lysine decarboxylase which converts lysine to cadaverine. In the absence of appropriate oral hygiene, overgrowth of these bacteria depletes lysine. This may contribute to gingival inflammation, while cadaverine contributes to oral malodor. A selective and sensitive capillary electrophoresis method with laser-induced fluorescence detection has been developed for the determination of cadaverine and lysine in saliva, as an indicator of lysine decarboxylase enzyme activity. The diamino compounds were separated in acidic background electrolyte in their mono-labeled form after derivatization with 4-fluoro-7-nitrobenz-2-oxa-1,3-diazole (NBD-F). Linearity and reproducibility of the method in the range 1-50 ?mol L(-1) have been demonstrated using saliva samples. The method was applied for the measurement of cadaverine and lysine in the saliva of healthy volunteers with or without proper oral hygiene. In the absence of oral hygiene, the mol fraction of cadaverine to cadaverine plus lysine in saliva increased significantly (0.65?±?0.13 vs. 0.39?±?0.18, P?

Tábi, Tamás; Lohinai, Zsolt; Pálfi, Melinda; Levine, Martin; Szöko, Eva

2008-05-01

307

Conjugation of ?-amylase with dextran for enhanced stability: process details, kinetics and structural analysis.  

PubMed

The influence of enzyme polysaccharide interaction on enzyme stability and activity was elucidated by covalently binding dextran to a model enzyme, ?-amylase. The conjugation process was optimized with respect to concentration of oxidizing agent, pH of enzyme solution, ratio of dextran to enzyme concentration, temperature and time of conjugate formation, and was found to affect the stability of ?-amylase. ?-Amylase conjugated under optimized conditions showed 5% loss of activity but with enhanced thermal and pH stability. Lower inactivation rate constant of conjugated ?-amylase within the temperature range of 60-80 °C implied its better stability. Activation energy for denaturation of ?-amylase increased by 8.81 kJ/mol on conjugation with dextran. Analysis of secondary structure of ?-amylase after covalent binding with dextran showed helix to turn conversion without loss of functional properties of ?-amylase. Covalent bonding was found to be mandatory for the formation of conjugate. PMID:22944451

Jadhav, Swati B; Singhal, Rekha S

2012-11-01

308

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

PubMed Central

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

Everill, B.; Berry, M. S.

1995-01-01

309

Visual tests for urinary amylase investigated in the routine laboratory.  

PubMed

Ninety-seven fresh urine specimens were tested in the routine laboratory with the Rapignost Amylase test strip, and the results were compared to those obtained using Phadebas Amylase test tablets to investigate the transferability of the results obtained by the Rapignost method to those of the Phadebas method under routine conditions. The fraction of conflicting negative results of Phadebas-positive specimens was 9% and the corresponding fraction of conflicting positive results was 13%. An attempt to improve the transferability by changing the comparison scale slightly did not succeed. However, a visual binary test based on 180 s incubation at 37 degrees C of 400 microliters urine in a suspension of a Phadebas Amylase test tablet seemed more suitable in selection of specimens with Phadebas amylase activity less than 2000 U/l (upper limit of the reference interval). These specimens amounted to approximately 75% of all amylase specimens in our laboratory. PMID:2408318

Uldall, A

1985-04-01

310

Amylase Synthesis and Stability in Crested Wheatgrass Seeds at Low Water Potentials 1  

PubMed Central

Drying of seeds of Agropyron desertorum (Fisch. ex Link) Schult. did not result in breakdown of ?-amylase nor impair the ability of seeds to resume its synthesis when moistened again. ?-Amylase activity did not change during 5 days of germination at a water potential of 0 atmosphere nor during 40 days of incubation at ?40 atmospheres. Seeds synthesized ?-amylase at 0, ?20, and ?40 atmospheres, but not at ?60 atmospheres. At 0 and ?20 atmospheres, the log of ?-amylase activity was linearly related to hastening of germination. But at ?40 atmospheres, seeds synthesized ?-amylase during a time when there was little hastening of germination. Thus, it appears that other biochemical reactions are less drought-tolerant than synthesis of ?-amylase. It is concluded that inhibition of ?-amylase synthesis is not a controlling factor in the germination of these seeds at low water potentials.

Wilson, A. M.

1971-01-01

311

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

PubMed Central

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

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

2013-01-01

312

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

PubMed

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

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

2013-01-01

313

What interactions drive the salivary mucosal pellicle formation?  

PubMed

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

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

2014-08-01

314

What interactions drive the salivary mucosal pellicle formation?  

PubMed Central

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

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

2014-01-01

315

beta-Adrenergic-responsive activation of extracellular signal-regulated protein kinases in salivary cells: role of epidermal growth factor receptor and cAMP.  

PubMed

The beta-adrenergic receptor agonist isoproterenol exerts growth-promoting effects on salivary glands. In this study, activation of ERKs, members of the mitogen-activated protein kinase family, by isoproterenol was examined in a human salivary gland cell line (HSY). Immunoblot analysis indicated that isoproterenol (10(-5) M) induced transient activation of ERK1/2 (4.4-fold relative to basal at 10 min) similar to that caused by EGF (6.7 fold). Isoproterenol, like EGF, also induced phosphorylation of the EGF receptor. However, inhibition of EGF receptor phosphorylation by the tyrphostin AG-1478 only partially attenuated isoproterenol-induced ERK phosphorylation, whereas EGF-responsive ERK activation was completely blocked. The G(i) inhibitor pertussis toxin also caused partial inhibition of isoproterenol-stimulated ERK activation. The cAMP analog 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate (CPT-cAMP) and the cAMP-elevating agents IBMX and cholera toxin produced transient ERK1/2 activation, similar to the effect of isoproterenol, in HSY cells. The stimulatory effects of isoproterenol and cAMP on ERK phosphorylation were not reduced by the PKA inhibitor H-89, whereas the Src family inhibitor 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidase (PP2) and transfection of a dominant-negative Src construct diminished isoproterenol-induced ERK activation. Isoproterenol induced marked overexpression of the cell growth-related adhesion molecule CD44, and this effect of isoproterenol was abolished by the ERK pathway inhibitor PD-98059. In summary, we show a dual mechanism of isoproterenol-induced ERK phosphorylation in HSY cells-one pathway mediated by EGF receptor transactivation and the other by an EGF receptor-independent pathway possibly mediated by cAMP. Our results also suggest that isoproterenol-induced growth of salivary tissue may involve ERK-mediated CD44 expression. PMID:15689414

Yeh, Chih-Ko; Ghosh, Paramita M; Dang, Howard; Liu, Qun; Lin, Alan L; Zhang, Bin-Xian; Katz, Michael S

2005-06-01

316

Production of Desmodus rotundus salivary plasminogen activator alpha1 (DSPAalpha1) in tobacco is hampered by proteolysis.  

PubMed

The high fibrin specificity of Desmodus rotundus salivary plasminogen activator alpha1 (DSPAalpha1 or desmoteplase (INN)) makes it a promising candidate for the treatment of acute ischemic stroke. In the current study we explored the use of transgenic tobacco plants and BY-2 suspension cells as alternative production platforms for this drug. Four different N-terminal signal peptides, from plants and animals, were used to translocate the recombinant DSPAalpha1 protein to the endomembrane system. Intact recombinant DSPAalpha1 was produced in transgenic plants and BY-2 cells, although a certain degree of degradation was observed in immunoblotted extracts. The choice of signal peptide had no major influence on the degradation pattern or recombinant protein accumulation, which reached a maximum level of 38 microg/g leaf material. N-terminal sequencing of purified, His6-tagged DSPAalpha1 revealed only minor changes in the position of signal peptide cleavage compared to the same protein expressed in Chinese hamster ovary cells. However, correctly processed recombinant DSPAalpha1 was also detected. The enzymatic activity of the recombinant protein was confirmed using an in vitro assay with unpurified and purified samples, demonstrating that plants are suitable for the production of functional DSPAalpha1. In contrast to whole plant cell extracts, no recombinant DSPAalpha1 was detected in the culture supernatant of transgenic BY-2 cells. Further analysis showed that recombinant DSPAalpha1 is subject to proteolysis and that endogenous secreted BY-2 proteases are responsible for DSPAalpha1 degradation in the culture medium. The addition of a highly concentrated protease inhibitor mixture or 5 mM EDTA reduced DSPAalpha1 proteolysis, improving the accumulation of intact product in the culture medium. Strategies to improve the plant cell suspension system for the production of secreted recombinant proteins are discussed. PMID:15685597

Schiermeyer, Andreas; Schinkel, Helga; Apel, Stefanie; Fischer, Rainer; Schillberg, Stefan

2005-03-30

317

[Immobilization of alpha-amylase on porous glass and silochrome].  

PubMed

Immobilized forms of alpha-amylase from Aspergillus oryzae were prepared on the porous glass and silochrome by the glutaraldehyde method. An addition of calcium ions at a concentration of 0.05 M to the reaction mixture during immobilization stabilized the enzyme activity. pH optimum of the insoluble form of alpha-amylase was 5.8 and that of the soluble form was 4.7. Storage of the insoluble enzyme as water suspension in 0.015 M CaCl2 at 4 degrees C for six months and twenty times repeated specific reaction did not affect significantly the activity of insoluble alpha-amylase. PMID:24839

Dvali, M Sh; Varlamov, V P; Kvesitadze, G I; Rogozhin, S V

1978-01-01

318

Comparison of the ?-Amylase of Bacillus subtilis and Bacillus amyloliquefaciens  

PubMed Central

The ?-amylase (?-1,4-glucan 4-glucanohydrolase, EC 3.2.1.1) of Bacillus subtilis strain W23 is less negatively-charged than the ?-amylase of B. amyloliquefaciens strain F, as determined by electrophoretic mobility in polyacrylamide gel at pH 8.6. The ?-amylase of strain W23 is immunologically unrelated to the ?-amylase of strain F, as judged by lack of cross-reaction in Ouchterlony immunodiffusion studies. The pH range of maximal activity for the enzyme of strain W23 was 5.7 to 6.7, with a maximum at 6.3. The pH range of activity for the ?-amylase of strain F was 5.5 to 6.5, with a maximum at 5.9. No significant difference was found in the effect of temperature on the activity of the ?-amylase of strain W23 and strain F. ?-Amylase production by strain W23 occurs throughout the 7-hr growth period, whereas enzyme production by strain F does not begin until the culture enters the stationary phase of growth. The total amounts of enzyme produced by strains W23 and F after 7 hr of growth were 0.3 and 25.5 units/ml, respectively. Images

Welker, N. E.; Campbell, L. Leon

1967-01-01

319

Salivary alpha amylase–cortisol asymmetry in maltreated youth  

Microsoft Academic Search

BackgroundMaltreatment represents a major stressor in the lives of many youth. Given the known effects of stress exposure on subsequent functioning of biological stress response systems, researchers have been interested in the effects of maltreatment on the functioning of these systems. Experimental studies reveal that previous exposure to stress affects the symmetry between components of the physiological stress response to

Elana B. Gordis; Douglas A. Granger; Elizabeth J. Susman; Penelope K. Trickett

2008-01-01

320

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

Microsoft Academic Search

We examined relations among cortisol, markers of sympathetic nervous system (SNS) activity (including salivary alpha-amylase\\u000a 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 would be associated with internalizing and externalizing problems.\\u000a Saliva samples were obtained from 8-

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

2008-01-01

321

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

PubMed Central

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

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

2013-01-01

322

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

PubMed

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

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

2009-08-01

323

Purification and determination of the NH2-terminal amino acid sequence of mouse alpha-amylase secreted from Saccharomyces cerevisiae: correct processing of the secretion signal from pGKL killer 28 kDa precursor protein.  

PubMed

We have previously reported the construction of recombinant mouse salivary alpha-amylase secretion vector in Saccharomyces cerevisiae utilizing novel yeast secretion signal derived from killer 28 kDa precursor protein. Here, we have first purified recombinant mouse alpha-amylase to homogeneity from the culture medium of S. cerevisiae, and determined its NH2-terminal amino acid sequence. The sequencing data indicated that the 28 kDa killer secretion signal-alpha-amylase fusion protein was cleaved accurately at its native processing site, and that both the core-glycosylated and non-glycosylated alpha-amylases possessed the same NH2-terminal amino acid sequences. PMID:1932087

Tokunaga, M; Kawamura, A; Omori, A; Hishinuma, F

1991-10-25

324

Deletion of ATG5 shows a role of autophagy in salivary homeostatic control.  

PubMed

Autophagy is a catabolic pathway utilized to maintain a balance among the synthesis, degradation, and recycling of cellular components, thereby playing a role in cell growth, development, and homeostasis. Previous studies revealed that a conditional knockout of essential member(s) of autophagy in a variety of tissues causes changes in structure and function of these tissues. Acinar cell-specific expression of knocked-in Cre recombinase through control of aquaporin 5 (Aqp5) promoter/enhancer (Aqp5-Cre) allows us to specifically inactivate Atg5, a protein necessary for autophagy, in salivary acinar cells of Atg5(f/f);Aqp5-Cre mice. There was no difference in apoptotic or proliferation levels in salivary glands of Atg5/Cre mice from each genotype. However, H&E staining and electron microscopy studies revealed modestly enlarged acinar cells and accumulated secretory granules in salivary glands of Atg5(f/f);Aqp5-Cre mice. Salivary flow rates and amylase contents of Atg5/Cre mice indicated that acinar-specific inactivation of ATG5 did not alter carbachol-evoked saliva and amylase secretion. Conversely, autophagy intersected with salivary morphological and secretory manifestations induced by isoproterenol administration. These results identified a role for autophagy as a homeostasis control in salivary glands. Collectively, Atg5(f/f);Aqp5-Cre mice would be a useful tool to enhance our understanding of autophagy in adaptive responses following targeted head and neck radiation or Sjögren syndrome. PMID:23884556

Morgan-Bathke, M; Lin, H H; Chibly, A M; Zhang, W; Sun, X; Chen, C-H; Flodby, P; Borok, Z; Wu, R; Arnett, D; Klein, R R; Ann, D K; Limesand, K H

2013-10-01

325

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

PubMed Central

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

Germaine, G R; Tellefson, L M

1986-01-01

326

Control of amylase production and growth characteristics of Aspergillus ochraceus.  

PubMed

The growth and the extracellular amylase production by Aspergillus ochraceus were studied in a stationary culture medium. Maximum growth rate of this fungus was found after 5 days of incubation at 30 degrees C, but maximum amylase production was obtained after 2 days. The highest amylase production were attained with lactose, maltose, xylose and starch as carbon sources. The extracellular amylase production and mycelial growth were influenced by the concentration of starch. Other carbohydrates supported growth but did not induce amylase synthesis and glucose repressed it, indicating catabolite repression in this microorganism. The presence of both mechanisms of induction and repression suggests that at least these multiple forms of regulation are present in A. ochraceus. Of the nitrogen sources tested, casaminoacids, ammonium nitrate and sodium nitrate stimulated the highest yield of amylase. Optimal amylase production was obtained at pH 5.0, but enzyme activity was found only in the 4.0-6.0 pH range. These results were probably due to the inhibitory effect of NH4(+)-N in the culture medium. PMID:17061508

Nahas, Ely; Waldemarin, Mirela M

2002-01-01

327

Modulation of inhibitory activity of xylanase - ?-amylase inhibitor protein (XAIP): binding studies and crystal structure determination of XAIP- II from Scadoxus multiflorus at 1.2 Ĺ resolution  

Microsoft Academic Search

BACKGROUND: Plants produce a wide range of proteinaceous inhibitors to protect themselves against hydrolytic enzymes. Recently a novel protein XAIP belonging to a new sub-family (GH18C) was reported to inhibit two structurally unrelated enzymes xylanase GH11 and ?-amylase GH13. It was shown to inhibit xylanase GH11 with greater potency than that of ?-amylase GH13. A new form of XAIP (XAIP-II)

Sanjit Kumar; Nagendra Singh; Biswajit Mishra; Divya Dube; Mau Sinha; S Baskar Singh; Sharmistha Dey; Punit Kaur; Sujata Sharma; Tej P Singh

2010-01-01

328

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

PubMed Central

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.

Soyk, Sebastian; Simkova, Klara; Zurcher, Evelyne; Luginbuhl, Leonie; Brand, Luise H.; Vaughan, Cara K.; Wanke, Dierk; Zeeman, Samuel C.

2014-01-01

329

Regulation of salivary-gland-specific gene expression.  

PubMed

The results from in vivo transgenic and in vitro transfection studies designed to identify cis-element(s) and transfactor(s) governing the salivary proline-rich proteins (PRPs), amylase, and parotid secretory protein (PSP) gene expression are utilized as a paradigm to discuss the regulation of salivary-specific gene expression. Particular attention is given to the molecular mechanism(s) underlying the salivary PRP R15 gene regulation. In rodents, the PRPs are selectively expressed in the acinar cells of salivary glands, and are inducible by the beta-agonist isoproterenol and by dietary tannins. The results from a series of experiments using chimeric reporter constructs containing different lengths of the R15 distal enhancer region, their mutations, and various expressing constructs are analyzed and discussed. These data suggest that the inducible nuclear orphan receptor NGFI-B may participate in the regulation of salivary acinar-cell-specific and inducible expression of the rat R15 gene via three distinct distal NGFI-B sites. Taken together, a model for the induction of R15 gene expression by Ipr is proposed. However, the exact molecular basis of this NGFI-B-mediated transactivation of cAMP-regulated R15 expression remains to be established. PMID:9260042

Ann, D K; Lin, H H; Kousvelari, E

1997-01-01

330

Extracellular amylase(s) production by fungi Botryodiplodia theobromae and Rhizopus oryzae grown on cassava starch residue.  

PubMed

The fungi Botryodiplodia theobromae and Rhizopus oryzae produce extracellular amylase when grown on a liquid medium containing 2% (WN) soluble starch or cassava starch residue(CSR) (as starch equivalent), a waste generated after extraction of starch from cassava, as the sole carbon source. Using CSR as the sole carbon source, the highest amylase activity of 3.25 and 3.8 units (mg, glucose released x ml(-1) x h(-1)) were obtained in shake flask cultures during the late stationary phase of growth of B. theobromae and R. oryzae, respectively. These values were slightly lower than the values obtained using soluble starch as the carbon source. Maximum enzyme synthesis in CSR incorporated medium occurred at the growth temperature of 30 degrees C and pH 6.0. Presence of inorganic NH4+ salts like ammonium acetate and ammonium nitrate in culture medium yielded more amylase than the other nitrogen sources. Amylase(s) production in the controlled environment of a Table-Top glass Jar Fermenter (2-L capacity) was 4.8 and 5.1 units for B. theobromae and R. oryzae, respectively using CSR as the carbon substrate. It is concluded that CSR, a cheap agricultural waste obtained after starch extraction from cassava could replace soluble starch as carbon substrate for commercial production of fungal amylase(s). PMID:15907080

Ray, R C

2004-10-01

331

Mapping of sequence-specific markers and loci controlling preharvest sprouting and alpha-amylase activity in rye ( Secale cereale L.) on the genetic map of an F 2 (S120×S76) population  

Microsoft Academic Search

Location of the loci that control preharvest sprouting and alpha-amylase activity in rye was studied based on intercross S120×S76,\\u000a consisting of 110 genotypes of F2 and F3 progenies. The genetic map currently consists of 141 loci distributed in 11 linkage groups, covering a distance of 506.4\\u000a cM, and was enriched during this study with 24 sequence-specific markers (7 SCARs, 7

B. My?eków; S. Stoja?owski; P. Milczarski; P. Masoj?

2010-01-01

332

Voltage and Ca2+-activated K+ channel in baso-lateral acinar cell membranes of mammalian salivary glands  

Microsoft Academic Search

Nervous or hormonal stimulation of many exocrine glands evokes release of cellular K+ (ref. 1), as originally demonstrated in mammalian salivary glands2,3, and is associated with a marked increase in membrane conductance1,4,5. We now demonstrate directly, by using the patch-clamp technique6, the existence of a K+ channel with a large conductance localized in the basolateral plasma membranes of mouse and

Y. Maruyama; D. V. Gallacher; O. H. Petersen

1983-01-01

333

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

Microsoft Academic Search

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

Rodrigo A. F. Redondo; Fabrício R. Santos

2006-01-01

334

Botulinum toxin A inhibits salivary secretion of rabbit submandibular gland.  

PubMed

Botulinum toxin A (BTXA) has been used in several clinical trials to treat excessive glandular secretion; however, the precise mechanism of its action on the secretory function of salivary gland has not been fully elucidated. In this study, we aimed to investigate the effect of BTXA on secretion of submandibular gland in rabbits and to identify its mechanism of action on the secretory function of salivary gland. At 12 weeks after injection with 5 units of BTXA, we found a significant decrease in the saliva flow from submandibular glands, while the salivary amylase concentration increased. Morphological analysis revealed reduction in the size of acinar cells with intracellular accumulation of secretory granules that coalesced to form a large ovoid structure. Expression of M3-muscarinic acetylcholine receptor (M3 receptor) and aquaporin-5 (AQP5) mRNA decreased after BTXA treatment, and distribution of AQP5 in the apical membrane was reduced at 1, 2 and 4 weeks after BTXA injection. Furthermore, BTXA injection was found to induce apoptosis of acini. These results indicate that BTXA decreases the fluid secretion of submandibular glands and increases the concentration of amylase in saliva. Decreased expression of M3 receptor and AQP5, inhibition of AQP5 translocation, and cell apoptosis might involve in BTXA-reduced fluid secretion of submandibular glands. PMID:24158141

Shan, Xiao-Feng; Xu, Hui; Cai, Zhi-Gang; Wu, Li-Ling; Yu, Guang-Yan

2013-12-01

335

Botulinum toxin A inhibits salivary secretion of rabbit submandibular gland  

PubMed Central

Botulinum toxin A (BTXA) has been used in several clinical trials to treat excessive glandular secretion; however, the precise mechanism of its action on the secretory function of salivary gland has not been fully elucidated. In this study, we aimed to investigate the effect of BTXA on secretion of submandibular gland in rabbits and to identify its mechanism of action on the secretory function of salivary gland. At 12 weeks after injection with 5 units of BTXA, we found a significant decrease in the saliva flow from submandibular glands, while the salivary amylase concentration increased. Morphological analysis revealed reduction in the size of acinar cells with intracellular accumulation of secretory granules that coalesced to form a large ovoid structure. Expression of M3-muscarinic acetylcholine receptor (M3 receptor) and aquaporin-5 (AQP5) mRNA decreased after BTXA treatment, and distribution of AQP5 in the apical membrane was reduced at 1, 2 and 4 weeks after BTXA injection. Furthermore, BTXA injection was found to induce apoptosis of acini. These results indicate that BTXA decreases the fluid secretion of submandibular glands and increases the concentration of amylase in saliva. Decreased expression of M3 receptor and AQP5, inhibition of AQP5 translocation, and cell apoptosis might involve in BTXA-reduced fluid secretion of submandibular glands.

Shan, Xiao-Feng; Xu, Hui; Cai, Zhi-Gang; Wu, Li-Ling; Yu, Guang-Yan

2013-01-01

336

Multisubstrate specifics amylase from mushroom Termitomyces clypeatus  

Microsoft Academic Search

An amylase was purified from the culture filtrate ofTermitomyces clypeatus by ammonium sulphate precipitation, DEAE-Sephadex chromatography and gel filtration on Bio-Gel P-200 column. The electrophoretically\\u000a homogeneous preparation also exhibited hydrolytic activity (in a decreasing order) on amylose, xylan, amylopectin, glycogen,\\u000a arabinogalactan and arabinoxylan. The enzyme had characteristically endo-hydrolytic activity on all the substrates tested\\u000a and no xylose, glucose, arabinose or

Anil K. Ghosh; Subhabrata SenGupta

1987-01-01

337

Involvement of nuclear orphan receptor NGFI-B in transcriptional activation of salivary-specific R15 gene by cAMP.  

PubMed

Proline-rich proteins (PRPs) are selectively expressed in the acinar cells of the salivary glands and are inducible by beta-agonist isoproterenol and dietary tannins. In the previous studies of rat PRP gene, R15, the 5'-flanking region up to -1.7 kilobase pairs (kb), which was thought to contain the necessary proximal regulatory elements, failed to confer the catecholamine isoproterenol and dietary tannin inducibility to the transgene expression in the salivary glands of transgenic mice. Here we analyzed distal 5'-flanking region of R15 in order to understand the mechanisms of tissue-specific and inducible gene regulation. An upstream regulatory region located between -2.4 and -1.7 kb of the R15 5'-flanking region is demonstrated to be indispensable for the salivary-specific and inducible reporter gene expression in vivo, by transgenic approach. Element(s) within the 0.7-kb (-2.4 to -1.7) region that is able to cis-activate the expression of a heterologous reporter gene expression is further elucidated by transient transfection assays in vitro. Three distinct nuclear orphan receptor NGFI-B regulatory sequences are identified within a 184-base pair (bp) minimal control region extended from -1995 to -1812 nucleotides relative to the transcription start site. When reporter gene containing this 184-bp control region and heterologous promoter was cotransfected with the NGFI-B expression construct, a transactivation that mimics the effect of cAMP is observed in the parotid cells. Finally, mutations on all three identified NGFI-B binding sites and coexpression of a dominant negative mutant construct, pCMV-NGFI-B(Delta25-195), abolish this transactivation mediated by NGFI-B. In summary, these data suggest that the inducible nuclear orphan receptor NGFI-B may participate in the regulation of salivary acinar cell-specific and inducible expression of the rat R15 gene via three distinct distal NGFI-B sites. PMID:8910353

Lin, H H; Tu, Z J; Ann, D K

1996-11-01

338

[Production of immobilized alpha-amylase and its properties].  

PubMed

Active immobilized alpha-amylase was obtained when applying AE-cellulose chromatography and glutaric dialdehyde. The time of the enzyme interaction with the carrier, amount of glutaric dialdehyde necessary for binding, optimal enzyme: carrier ratio as well as the methods for desiccation of the immobilized amylase preparations were specified. Conditions are selected for alpha-amylase stabilization with the presence of glutaric aldehyde. Immobilized amylase as compared to free enzyme is shown to be more pH-stable in the acid and alkaline zones of pH (2.0-3.5 and 10.5-12.0), thermostable (within a temperature range of 20-60 degrees C) and resistant to the effect of 5.5 M urea. PMID:982618

Galich, I P; Tsyperovich, A S; Kolesnik, L A; Tsesarskaia, V D

1976-01-01

339

Enzymatic detergent formulation containing amylase from Aspergillus niger: A comparative study with commercial detergent formulations  

Microsoft Academic Search

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 (3.9Uml?1±0.2) in submerged culture and its amylase demonstrated excellent activity

Sydnei Mitidieri; Anne Helene Souza Martinelli; Augusto Schrank; Marilene Henning Vainstein

2006-01-01

340

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

PubMed

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

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

1991-07-01

341

Salivary gland determination in Drosophila: a salivary-specific, fork head enhancer integrates spatial pattern and allows fork head autoregulation.  

PubMed

In the early Drosophila embryo, a system of coordinates is laid down by segmentation genes and dorsoventral patterning genes. Subsequently, these coordinates must be interpreted to define particular tissues and organs. To begin understanding this process for a single organ, we have studied how one of the first salivary gland genes, fork head (fkh), is turned on in the primordium of this organ, the salivary placode. A placode-specific fkh enhancer was identified 10 kb from the coding sequence. Dissection of this enhancer showed that the apparently homogeneous placode is actually composed of at least four overlapping domains. These domains appear to be developmentally important because they predict the order of salivary invagination, are evolutionarily conserved, and are regulated by patterning genes that are important for salivary development. Three dorsoventral domains are defined by EGF receptor (EGFR) signaling, while stripes located at the anterior and posterior edges of the placode depend on wingless signaling. Further analysis identified sites in the enhancer that respond either positively to the primary activator of salivary gland genes, SEX COMBS REDUCED (SCR), or negatively to EGFR signaling. These results show that fkh integrates spatial pattern directly, without reference to other early salivary gland genes. In addition, we identified a binding site for FKH protein that appears to act in fkh autoregulation, keeping the gene active after SCR has disappeared from the placode. This autoregulation may explain how the salivary gland maintains its identity after the organ is established. Although the fkh enhancer integrates information needed to define the salivary placode, and although fkh mutants have the most extreme effects on salivary gland development thus far described, we argue that fkh is not a selector gene for salivary gland development and that there is no master, salivary gland selector gene. Instead, several genes independently sense spatial information and cooperate to define the salivary placode. PMID:11518505

Zhou, B; Bagri, A; Beckendorf, S K

2001-09-01

342

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

PubMed

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

Saha, Koel; Maity, Sujan; Roy, Sudeshna; Pahan, Koustav; Pathak, Rishija; Majumdar, Susmita; Gupta, Suvroma

2014-01-01

343

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

PubMed Central

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

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

2014-01-01

344

Biochemical properties of alpha-amylase from peel of Citrus sinensis cv. Abosora.  

PubMed

alpha-Amylase activity was screened in the peel, as waste fruit, of 13 species and cultivars of Egyptian citrus. The species Citrus sinensis cv. Abosora had the highest activity. alpha-Amylase AI from Abosora peel was purified to homogeneity using anion and cation-exchange, and gel filtration chromatographies. Molecular weight of alpha-amylase AI was found to be 42 kDa. The hydrolysis properties of alpha-amylase AI toward different substrates indicated that corn starch is the best substrate. The alpha-amylase had the highest activity toward glycogen compared with amylopectin and dextrin. Potato starch had low affinity toward alpha-amylase AI but it did not hydrolyze beta-cyclodextrin and dextran. Apparent Km for alpha-amylase AI was 5 mg (0.5%) starch/ml. alpha-Amylase AI showed optimum activity at pH 5.6 and 40 degrees C. The enzyme was thermally stable up to 40 degrees C and inactivated at 70 degrees C. The effect of mono and divalent metal ions were tested for the alpha-amylase AI. Ba2+ was found to have activating effect, where as Li+ had negligible effect on activity. The other metals caused inhibition effect. Activity of the alpha-amylase AI was increased one and half in the presence of 4 mM Ca2+ and was found to be partially inactivated at 10 mM Ca2+. The reduction of starch viscosity indicated that the enzyme is endoamylase. The results suggested that, in addition to citrus peel is a rich source of pectins and flavanoids, alpha-amylase AI from orange peel could be involved in the development and ripening of citrus fruit and may be used for juice processing. PMID:19941088

Mohamed, Saleh Ahmed; Drees, Ehab A; El-Badry, Mohamed O; Fahmy, Afaf S

2010-04-01

345

The effect of concentration of tannin-rich bean hulls (Vicia faba L.) on activities of lipase (EC 3.1.1.3) and alpha-amylase (EC 3.2.1.1) in digesta and pancreas and on the digestion of lipid and starch by young chicks.  

PubMed

The effect of different concentrations of tannin-rich field-bean (Vicia faba L.) hulls at 0, 20, 50, 150 and 300 g/kg dietary inclusion on the activities of lipase (EC 3.1.1.3) and alpha-amylase (EC 3.2.1.1) in digesta and pancreas and on the digestion of lipid and starch was studied in 2-3-week-old male broiler chicks. Low dietary concentrations of tannins (20 and 50 g hulls/kg) enhanced the activity of lipase in digesta from both the jejunum and ileum, the 20 g hulls/kg effecting the greatest enhancement, but no stimulatory effect on the activity of digesta alpha-amylase was observed. High dietary concentrations of tannins (150 and 300 g hulls/kg) inhibited both lipase and alpha-amylase activities in digesta from both the jejunum and ileum, the 30 g hulls/kg causing the most inhibition. Tannins did not increase the activities of lipase or alpha-amylase in pancreatic homogenates, but at high concentrations (150 and 300 g hulls/kg) they lowered slightly the pancreatic activity of alpha-amylase. Nutrient digestion was less influenced by the concentration of tannins than digesta enzyme activities. PMID:1931901

Longstaff, M A; McNab, J M

1991-07-01

346

Starch deposition and amylase accumulation during somatic embryogenesis in bamboo (Dendrocalamus hamiltonii).  

PubMed

In monocots, the zygotic embryo is protected and nourished by an endosperm. In the present study starch deposition and amylase accumulation was noticed during somatic embryogenesis in stem callus of a bamboo, Dendrocalamus hamiltonii. SEM studies revealed that starch grains were clearly visible in the scutellum during the maturation stage of the somatic embryo. As the somatic embryo developed further, the scutellum got reduced with corresponding increase in amylase. The amylase activity was tested periodically at different developmental stages of embryos. The role of scutellum in somatic embryos for starch deposition and amylase accumulation is discussed. PMID:15022840

Godbole, Savita; Sood, Anil; Sharma, Madhu; Nagar, Pramod Kumar; Ahuja, Paramvir Singh

2004-02-01

347

Tumours of salivary tissue  

PubMed Central

A clinical study of 401 cases of tumour of salivary tissue has been made and the results reported, together with a review of the literature. The histological variations of such tumours are classified and discussed. Images

Cameron, J. Malcolm

1961-01-01

348

Salivary gland biopsy  

MedlinePLUS

... 21. Lacey J.Diagnostic Imaging and Fine-Needle Aspiration of the Salivary Glands. In: Flint PW, Haughey BH, Lund LJ, et al, eds. Cummings Otolaryngology: Head & Neck Surgery. 5th ed. Philadelphia, ...

349

Analysis of the salivary gland transcriptome of Frankliniella occidentalis.  

PubMed

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

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

2014-01-01

350

Salivary and serum analysis in children diagnosed with pneumonia.  

PubMed

The aim of the current study was to evaluate specific markers for pneumonia by using a non-invasive assessment of inflammatory/oxidative biomarkers in saliva accompanying a routine serum analysis. No study evaluating saliva of children with pneumonia has been published previously. Salivary analysis was performed in 15 children diagnosed with lobar pneumonia and in a parallel group of 16 children matching in age and gender in whom there was no respiratory illness, and compared to the serum analysis obtained routinely in both groups of children. Salivary flow rate was lower in the patients' group as was uric acid concentration (by 60%). Increase in salivary concentrations of almost all parameters analyzed was found: Ca, P, and Mg concentrations were higher in the patients' group by 23%, 55%, and 33%, respectively, while LDH, total protein amylase and albumin concentrations were higher by 275%, 79%, and 42%, respectively. In the serum, white cell counts and neutrophils were significantly higher, and sodium level significantly lower in the patients' group. Compositional changes were in the range of 3-80% while the saliva alterations were more profound, in the range of 42-275%. The results demonstrated in the current study indicate salivary analysis as a potentially novel tool for children with pneumonia. Human salivary collection and analysis is a non-invasive tool that could provide additional information for diagnosis and follow-up of pneumonia, especially in children. This is especially beneficial for pediatric patients, as salivary collection is simple, non-invasive, and patient-friendly. Pediatr Pulmonol. 2014; 49:569-573. © 2013 Wiley Periodicals, Inc. PMID:23532916

Klein Kremer, Adi; Kuzminsky, Ela; Bentur, Lea; Nagler, Rafael M

2014-06-01

351

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

Microsoft Academic Search

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

Yvonne R. Carter; Michael Ashburner

1972-01-01

352

Deep metaproteomic analysis of human salivary supernatant  

PubMed Central

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

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

2012-01-01

353

Genetically engineered human salivary histatin genes are functional in Candida albicans: development of a new system for studying histatin candidacidal activity  

Microsoft Academic Search

Histatins are a structurally related family of salivary proteins known as histidine-rich proteins that are produced and secreted by the human major salivary glands. In vitro, histatins are potent cytotoxic proteins with selectivity for pathogenic yeasts including Candida albicans. Studies that investigate the mechanism of action of histatin proteins upon this important human pathogen have used a candidacidal assay in

Didi Baev; Xuewei Li; Mira Edgerton

354

Calibration of the Ektachem Amylase method using human reference materials and the Phadebas Blue Starch method as an interim reference method.  

PubMed

In a multilaboratory study a consensus was established to calibrate the Ektachem Amylase method to reproduce the results of the Phadebas Blue Starch method. The calibration graph has a slope = 3.39 and intercept = 25. For the Ektachem Amylase method the reaction-rate ratio between salivary and pancreatic amylase was calculated to be 0.89, relative to that of the Phadebas Blue Starch method. A calibration value for the Nordic Amylase Calibrator to be used on Ektachem analysers was determined to be close to 469 U l-1 for the current batch. However, since the difference from the stated value of 460 U l-1 is negligible, the authors recommend the use of the stated value for this and future batches of the Nordic Amylase Calibrator. An error of around 10% introduced by the presence of salivary amylase is comparable to the methods accepted by the Scandinavian Committee on Enzymes. Introduction of a consensus calibration reduced the interlaboratory variation by up to 40% at all levels. PMID:8903112

Mogensen, P K; Mřller, J; Nyborg, L; Magid, E; Wimmelmann, N; Uldall, A

1996-10-01

355

Further Experiments on Gibberellin-Stimulated Amylase Production in Cereal Grains  

ERIC Educational Resources Information Center

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

Coppage, Jo; Hill, T. A.

1973-01-01

356

Salivary Gland Cancer: Risk Factors  

MedlinePLUS

... Board , 4/2014 Risk Factors Cancer.Net Guide Salivary Gland Cancer Overview Statistics Medical Illustrations Risk Factors Symptoms ... and health care choices. The cause(s) of most salivary gland cancers are unknown, but the following factors may ...

357

Ingestion of potato starch decreases chymotrypsin but does not affect trypsin, amylase, or lipase activity in the pancreas in rats  

Microsoft Academic Search

Adaptation of digestive enzymes in the pancreas was evaluated in rats fed diets composed of potato starch as a carbohydrate source. Male Sprague-Dawley rats at 7 weeks were fed 4 different diets containing 60% sucrose, cornstarch, or 2 kinds of starch derived from different potato varieties. Enzyme activity in the pancreas of rats was determined at 0, 1, 3, and

Hitoshi Mineo; Kyo Ishida; Nao Morikawa; Sayako Ohmi; Ayaka Machida; Takahiro Noda; Michihiro Fukushima; Hideyuki Chiji

2007-01-01

358

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

PubMed Central

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

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

2013-01-01

359

Cloning and characterization of alpha-amylase from Atlantic salmon (Salmo salar L.).  

PubMed

Amylase has a lower activity in carnivorous fish species, particularly in Atlantic salmon. We report the first cloning of a salmonid alpha-amylase cDNA from Atlantic salmon, a major species in aquaculture. By amino acid alignment of several species, we identified a seven amino acid deletion in one of the large loops of the enzyme in relatively close proximity to the active site, that could impair substrate binding. We also found the signal peptide to be less hydrophobic compared to other species. This may affect import into ER during protein synthesis. Active site residues were shown to be conserved. Amylase mRNA expression was shown in pancreatic tissue, liver, and in the heart. Using blocked p-nitrophenyl-maltoheptaoside as a substrate, we measured a low amylase activity in Atlantic salmon intestinal content, which was about half of the activity measured in Atlantic cod, whereas activity measured in rainbow trout was fourteen times higher. Amylase activities in all three species showed similar degree of reduction in hydrolytic activity in a dose-response trial with a wheat amylase inhibitor preparation. This indicates similar specific activity per amylase molecule. PMID:17020811

Frřystad, Marianne K; Lilleeng, Einar; Sundby, Anne; Krogdahl, Ashild

2006-12-01

360

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

PubMed Central

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

Hamdan, Imad I.; Afifi, Fatima U.

2010-01-01

361

Inhibitory effects of chlorogenic acids from green coffee beans and cinnamate derivatives on the activity of porcine pancreas ?-amylase isozyme I  

Microsoft Academic Search

Nine kinds of chlorogenic acids (CGAs) account for 80% of the total CGA content in green coffee beans. They consist of three subgroups of caffeoylquinic acids (CQAs), feruloylquinic acids (FQAs), and dicaffeoylquinic acids (diCQAs). We previously reported the inhibitory effects of 5-CQA on porcine pancreas ?-amylase (PPA) isozymes, PPA-I and PPA-II. In this paper, we investigated the PPA-I inhibition by

Yusaku Narita; Kuniyo Inouye

2011-01-01

362

Analysis of salivary transcripts and antigens of the sand fly Phlebotomus arabicus  

PubMed Central

Background Sand fly saliva plays an important role in blood feeding and Leishmania transmission as it was shown to increase parasite virulence. On the other hand, immunity to salivary components impedes the establishment of infection. Therefore, it is most desirable to gain a deeper insight into the composition of saliva in sand fly species which serve as vectors of various forms of leishmaniases. In the present work, we focused on Phlebotomus (Adlerius) arabicus, which was recently shown to transmit Leishmania tropica, the causative agent of cutaneous leishmaniasis in Israel. Results A cDNA library from salivary glands of P. arabicus females was constructed and transcripts were sequenced and analyzed. The most abundant protein families identified were SP15-like proteins, ParSP25-like proteins, D7-related proteins, yellow-related proteins, PpSP32-like proteins, antigen 5-related proteins, and 34 kDa-like proteins. Sequences coding for apyrases, hyaluronidase and other putative secreted enzymes were also represented, including endonuclease, phospholipase, pyrophosphatase, amylase and trehalase. Mass spectrometry analysis confirmed the presence of 20 proteins predicted to be secreted in the salivary proteome. Humoral response of mice bitten by P. arabicus to salivary antigens was assessed and many salivary proteins were determined to be antigenic. Conclusion This transcriptomic analysis of P. arabicus salivary glands is the first description of salivary proteins of a sand fly in the subgenus Adlerius. Proteomic analysis of P. arabicus salivary glands produced the most comprehensive account in a single sand fly species to date. Detailed information and phylogenetic relationships of the salivary proteins are provided, expanding the knowledge base of molecules that are likely important factors of sand fly-host and sand fly-Leishmania interactions. Enzymatic and immunological investigations further demonstrate the value of functional transcriptomics in advancing biological and epidemiological research that can impact leishmaniasis.

Hostomska, Jitka; Volfova, Vera; Mu, Jianbing; Garfield, Mark; Rohousova, Iva; Volf, Petr; Valenzuela, Jesus G; Jochim, Ryan C

2009-01-01

363

PREFERENTIAL SYNTHESIS OF ?-AMYLASE BY BACILLUS STEAROTHERMOPHILUS IN THE PRESENCE OF 5-METHYL-TRYPTOPHAN1  

PubMed Central

Welker, N. E. (Western Reserve University, Cleveland, Ohio), and L. Leon Campbell. Preferential synthesis of ?-amylase by Bacillus stearothermophilus in the presence of 5-methyl-tryptophan. J. Bacteriol. 87:828–831. 1964.—Washed-cell suspensions of Bacillus stearothermophilus induced with pure maltose preferentially synthesized ?-amylase in the presence of 5-methyl-tryptophan (5-MT). 5-MT did not inhibit the formation of active ?-amylase or the incorporation of proline-C14 into ?-amylase. In contrast, p-fluorophenylalanine inhibited the formation of active ?-amylase by 92% and the incorporation of proline-C14 into the enzyme by 95%. p-Fluorophenylalanine and 5-MT inhibited cellular protein synthesis, as measured by proline-C14 incorporation, by 74 and 72%, respectively.

Welker, N. E.; Campbell, L. Leon

1964-01-01

364

Partial Purification and Characterization of a Thermostable Actinomycete ?-Amylase  

PubMed Central

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

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

1984-01-01

365

Diet and the evolution of human amylase gene copy number variation  

PubMed Central

Starch consumption is a prominent characteristic of agricultural societies and hunter-gatherers in arid environments. In contrast, rainforest and circum-arctic hunter-gatherers and some pastoralists consume much less starch1-3. 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 is correlated positively with salivary amylase protein levels, and that individuals from populations with high-starch diets have on average more AMY1 copies than those with traditionally low-starch diets. Comparisons with other loci in a subset of these populations suggest that the level of AMY1 copy number differentiation is unusual. This example of positive selection on a copy number variable gene is one of the first in the human genome. Higher AMY1 copy numbers and protein levels likely improve the digestion of starchy foods, and may buffer against the fitness-reducing effects of intestinal disease.

Perry, George H.; Dominy, Nathaniel J.; Claw, Katrina G.; Lee, Arthur S.; Fiegler, Heike; Redon, Richard; Werner, John; Villanea, Fernando A.; Mountain, Joanna L.; Misra, Rajeev; Carter, Nigel P.; Lee, Charles; Stone, Anne C.

2008-01-01

366

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

PubMed

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

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

2012-01-01

367

Radioisotope study of salivary glands  

SciTech Connect

The book discusses the use of radioisotope methods in the diagnosis of salivary gland diseases. Anatomical and physiological features of the salivary gland are summarized and radiotracer deposition processes are described. Clinical applications of scintigraphy are detailed. The degree of functional impairment due to various inflammatory diseases is contrasted by means of semiquantitative computerized methods with follow-up therapeutic results. Post-irradiatory involvement and possible functional recovery of salivary glands are also considered. The contents discussed are: Salivary Gland Physiology and Radioisotope Uptake. Radioisotope Study of Salivary Glands. Radioisotope Studies Under Normal Conditions. Survey of Radiographic Methods. Dosimetric Assessment. Conclusions and Index.

De Rossi, G.

1987-01-01

368

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

PubMed Central

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

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

1988-01-01

369

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

PubMed

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

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

1988-04-01

370

Multiple polyadenylation sites in a mouse alpha-amylase gene.  

PubMed

Two alpha-amylase mRNAs which differ in the length of their 3' non-translated region accumulate in the cytoplasm in both mouse liver and salivary gland tissues. The two species in each tissue are transcribed from the same gene (Amy-1A). The minor species is approximately 20-nucleotides preceding the poly(A) tract. Sequence analysis of genomic DNA shows that these extra 237 nucleotides are specified by sequences contiguous to those shared by the two mRNAs. These data demonstrate that transcription can proceed through the major polyadenylation site and that alternative polyadenylation sites are used in the Amy-1A gene. Sequences which trail the two polyadenylation sites exhibit extensive homology and might therefore be involved in polyadenylation or transcription termination. PMID:6166922

Tosi, M; Young, R A; Hagenbüchle, O; Schibler, U

1981-05-25

371

Pictorial essay: Salivary gland imaging  

PubMed Central

Salivary glands are the first organs of digestion secreting their digestive juices into the oral cavity. Parotid, submandibular, and sublingual glands are the major paired salivary glands in the decreasing order of their size. In addition, multiple small minor salivary glands are noted randomly distributed in the upper aerodigestive tract, including paranasal sinuses and parapharyngeal spaces. The imaging is directed to the major salivary glands. Commonly used imaging methods include plain radiography and conventional sialography. Recently, high-resolution ultrasonography (HRUS) is being increasingly used for targeted salivary gland imaging. However, the advent of cross-sectional imaging techniques such as computed tomography (CT) and magnetic resonance imaging (MRI) have revolutionized the imaging of salivary glands. This article illustrates the role of imaging in evaluating the variegated disease pattern of the major salivary glands.

Rastogi, Rajul; Bhargava, Sumeet; Mallarajapatna, Govindarajan Janardan; Singh, Sudhir Kumar

2012-01-01

372

Trastuzumab in Treating Patients With Metastatic or Recurrent Salivary Gland Cancer  

ClinicalTrials.gov

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

2013-02-27

373

Changes in functioning of rat submandibular salivary gland under streptozotocin-induced diabetes are associated with alterations of Ca2+ signaling and Ca2+ transporting pumps.  

PubMed

Xerostomia and pathological thirst are troublesome complications of diabetes mellitus associated with impaired functioning of salivary glands; however, their cellular mechanisms are not yet determined. Isolated acinar cells were loaded with Ca2+ indicators fura-2/AM for measuring cytosolic Ca2+ concentration ([Ca2+]i) or mag-fura-2/AM-inside the endoplasmic reticulum (ER). We found a dramatic decrease in pilocarpine-stimulated saliva flow, protein content and amylase activity in rats after 6 weeks of diabetes vs. healthy animals. This was accompanied with rise in resting [Ca2+]i and increased potency of acetylcholine (ACh) and carbachol (CCh) but not norepinephrine (NE) to induce [Ca2+]i transients in acinar cells from diabetic animals. However, [Ca2+]i transients mediated by Ca2+ release from ER stores (induced by application of either ACh, CCh, NE, or ionomycin in Ca2+-free extracellular medium) were decreased under diabetes. Application of inositol-1,4,5-trisphosphate led to smaller Ca2+ release from ER under the diabetes. Both plasmalemma and ER Ca2+-ATPases activity was reduced and the latter showed the increased affinity to ATP under the diabetes. We conclude that the diabetes caused impairment of salivary cells functions that, on the cellular level, associates with Ca2+ overload, increased Ca2+-mobilizing ability of muscarinic but not adrenergic receptors, decreased Ca2+-ATPases activity and ER Ca2+ content. PMID:16443349

Fedirko, N V; Kruglikov, I A; Kopach, O V; Vats, J A; Kostyuk, P G; Voitenko, N V

2006-03-01

374

Amylase creatinine clearance ratio after biliary surgery  

Microsoft Academic Search

The amylase creatinine clearance ratio (ACCR) is considered to be a more sensitive index of acute pancreatitis than the serum amylase level. Serial ACCR estimations were undertaken in 25 patients undergoing an elective cholecystectomy. Using accepted criteria, 28% of these patients developed, in the postoperative period, biochemical evidence of pancreatic gland damage, although the serum amylase level remained normal. This

L A Donaldson; W McIntosh; S N Joffe

1977-01-01

375

Salivary gland organogenesis.  

PubMed

Our understanding of vertebrate salivary gland organogenesis has been largely informed by the study of the developing mouse submandibular gland (SMG), which will be the major focus of this review. The mouse SMG has been historically used as a model system to study epithelial-mesenchymal interactions, growth factor-extracellular matrix (ECM) interactions, and branching morphogenesis. SMG organogenesis involves interactions between a variety of cell types and their stem/progenitor cells, including the epithelial, neuronal, and mesenchymal cells, and their ECM microenvironment, or niche. Here, we will review recent literature that provides conceptual advances in understanding the molecular mechanisms of salivary gland development. We will describe SMG organogenesis, introduce the model systems used to study development, and outline the key signaling pathways and cellular processes involved. We will also review recent research focusing on the identification of stem/progenitor cells in the SMG and how they are directed along a series of cell fate decisions to form a functional gland. The mechanisms that drive SMG organogenesis provide a template to regenerate functional salivary glands in patients who suffer from salivary hypofunction due to irreversible glandular damage after irradiation or removal of tumors. Additionally, these mechanisms may also control growth and development of other organ systems. PMID:23801668

Knosp, Wendy M; Knox, Sarah M; Hoffman, Matthew P

2012-01-01

376

Nucleotide sequence of the G6-amylase gene from alkalophilic Bacillus sp. H-167.  

PubMed

The nucleotide sequence of the G6-amylase gene from alkalophilic Bacillus sp. H-167 was determined. The open reading frame of the gene consisted of 2865 base pairs, encoding 955 amino acids. The NH2-terminal amino acid sequence analysis of the G6-amylase indicated that the enzyme had a single peptide of 33 amino acid residues and the mature enzyme was composed of 922 amino acids, giving a molecular mass of 102,598. Identity of the NH2-terminal amino acid sequences among each component of the multiform G6-amylase suggested the proteolytic processing of the COOH-terminal side of the enzyme. The DNA sequence and the deduced amino acid sequence of the G6-amylase gene showed no homology with those of other bacterial alpha-amylases although the consensus amino acid sequences of the active center were well conserved. PMID:2227348

Shirokizawa, O; Akiba, T; Horikoshi, K

1990-07-01

377

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

PubMed

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

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

2014-01-01

378

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

PubMed Central

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.

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

2014-01-01

379

Rab3D regulates amylase levels, not agonist-induced amylase release, in AR42J cells.  

PubMed

Rab3D is a low molecular weight GTP-binding protein that associates with secretory granules in exocrine cells. AR42J cells are derived from rat pancreatic exocrine tumor cells and develop an acinar cell-like phenotype when treated with dexamethasone (Dex). In the present study, we examined the role of Rab3D in Dex-treated AR42J cells. Rab3D expression and localization were analyzed by subcellular fractionation and immunoblotting. The role of Rab3D was examined by overexpressing myc-labeled wild-type-Rab3D and a constitutively active form of Rab3D (Rab3D-Q81L) in AR42J cells. We found that Rab3D is predominantly membrane-associated in AR42J cells and co-localizes with zymogen granules (ZG). Following CCK-8-induced exocytosis, amylase-positive ZGs appeared to move towards the periphery of the cell and co-localization between Rab3D and amylase was less complete when compared to basal conditions. Overexpression of WT, but not mutant Rab3D, resulted in an increase in cellular amylase levels. Overexpression of mutant and WT Rab3D did not affect granule morphology, CCK-8-induced secretion, long-term (48 hr) basal amylase release or granule density. We conclude that Rab3D is not involved in agonist-induced exocytosis in AR42J cells. Instead, Rab3D may regulate amylase content in these cells. PMID:22367855

Limi, Saima; Ojakian, George; Raffaniello, Robert

2012-06-01

380

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

Microsoft Academic Search

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

KARINE WILLEMOT; PIERRE CORNELIS

1983-01-01

381

Cloning and expression of Lipomyces starkeyi ?-amylase in Escherichia coli and determination of some of its properties  

Microsoft Academic Search

The Lipomyces starkeyi ?-amylase (LSA) gene encoding soluble starch-degrading ?-amylase was cloned and characterized from a derepressed and partially constitutive mutant for both dextranase and amylase activities. The nucleotide (nt) sequence of the cDNA fragment reveals an open reading frame of 1944 bp encoding a 619 amino acid (aa) mature protein (LSA) with a calculated molecular weight of 68.709 kDa

Hee Kyoung Kang; Jin Ha Lee; Doman Kim; Donal F. Day; John F. Robyt; Kwan-Hwa Park; Tae-Wha Moon

2004-01-01

382

Properties of an amylase from thermophilic Bacillus SP  

PubMed Central

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

de Carvalho, Raquel Vieira; Correa, Thamy Livia Ribeiro; da Silva, Julia Caroline Matos; de Oliveira Mansur, Luciana Ribeiro Coutinho; Martins, Meire Lelis Leal

2008-01-01

383

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

PubMed

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

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

2009-01-01

384

Temporal and compositional characteristics of salivary protein adsorption to hydroxyapatite.  

PubMed

Salivary proteins bind to enamel surfaces and hydroxyapatite in a highly selective manner. Numerous studies have identified these proteins as primarily proline-rich proteins, cystatins, statherin, and histatins. Previously, the hydroxyapatite-binding potential of these proteins had been characterized in systems consisting of singly purified protein and adsorbent. The purpose of this study was to investigate the adsorption of each protein in the presence of complete salivary secretion. Proteins, shown to adsorb to hydroxyapatite, were purified, biotinylated, and added back to the remaining proteins to form a series of reconstituted secretions. The adsorption of each biotinylated protein in the reconstituted secretion to hydroxyapatite was then measured as a function of time. Results indicated that three different adsorption patterns occur. A simple hyperbolic pattern is characteristic of amylase, glycosylated proline-rich protein (PRG), and cystatin. A faster adsorption process is observed for PRP-3, PRP-4, PIF-f, and statherin. A more complex pattern, exhibiting a rapid phase followed by a slower phase, is characteristic of PRP-1, PRP-2, PIF-s, and histatins. These results suggest that there are different adsorption processes involved in the binding of salivary proteins to hydroxyapatite. Two possible mechanisms are direct adsorption of protein to hydroxyapatite and indirect adsorption of protein by interacting with other proteins already bound to hydroxyapatite. PMID:8655778

Lamkin, M S; Arancillo, A A; Oppenheim, F G

1996-02-01

385

Crystal Structure of Bacillus subtilis ?-Amylase in Complex with Acarbose  

PubMed Central

The crystal structure of Bacillus subtilis ?-amylase, in complex with the pseudotetrasaccharide inhibitor acarbose, revealed an hexasaccharide in the active site as a result of transglycosylation. After comparison with the known structure of the catalytic-site mutant complexed with the native substrate maltopentaose, it is suggested that the present structure represents a mimic intermediate in the initial stage of the catalytic process.

Kagawa, Masayuki; Fujimoto, Zui; Momma, Mitsuru; Takase, Kenji; Mizuno, Hiroshi

2003-01-01

386

Organic solvent tolerance of halophilic a-amylase from a Haloarchaeon, Haloarcula sp. strain S-1  

Microsoft Academic Search

A halophilic archaeon, Haloarcula sp. strain S-1, produced extracellular organic solvent-tolerant ?-amylase. Molecular mass of the enzyme was estimated to be 70 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This amylase exhibited maximal activity at 50°C in buffer containing 4.3 M NaCl, pH 7.0. Moreover, the enzyme was active and stable in various organic solvents (benzene, toluene, and chloroform, etc.). Activity was not

Tadamasa Fukushima; Toru Mizuki; Akinobu Echigo; Akira Inoue; Ron Usami

2005-01-01

387

Salivary PYY: A Putative Bypass to Satiety  

PubMed Central

Peptide YY3-36 is a satiation hormone released postprandially into the bloodstream from L-endocrine cells in the gut epithelia. In the current report, we demonstrate PYY3-36 is also present in murine as well as in human saliva. In mice, salivary PYY3-36 derives from plasma and is also synthesized in the taste cells in taste buds of the tongue. Moreover, the cognate receptor Y2R is abundantly expressed in the basal layer of the progenitor cells of the tongue epithelia and von Ebner's gland. The acute augmentation of salivary PYY3-36 induced stronger satiation as demonstrated in feeding behavioral studies. The effect is mediated through the activation of the specific Y2 receptor expressed in the lingual epithelial cells. In a long-term study involving diet-induced obese (DIO) mice, a sustained increase in PYY3-36 was achieved using viral vector-mediated gene delivery targeting salivary glands. The chronic increase in salivary PYY3-36 resulted in a significant long-term reduction in food intake (FI) and body weight (BW). Thus this study provides evidence for new functions of the previously characterized gut peptide PYY3-36 suggesting a potential simple and efficient alternative therapeutic approach for the treatment of obesity.

Gorbatyuk, Oleg; La Sala, Michael; Duncan, David; Aslanidi, George; Campbell-Thompson, Martha; Zhang, Lei; Herzog, Herbert; Voutetakis, Antonis; Baum, Bruce J.; Zolotukhin, Sergei

2011-01-01

388

Taxonomy of Salivary Gland Neoplasm  

PubMed Central

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

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

2014-01-01

389

Basic fibroblast growth factor in rat salivary glands  

Microsoft Academic Search

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

Osamu Amano; Yoshino Yoshitake; Katsuzo Nishikawa; Shoichi Iseki

1993-01-01

390

Characteristics of Two Forms of ?-Amylases and Structural Implication  

PubMed Central

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

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

1999-01-01

391

Purification and characterization of digestive amylase from the tasar silkworm, Antheraea mylitta (Lepidoptera: Saturniidae)  

Microsoft Academic Search

Digestive amylase was purified from larvae of Indian tasar silkworm, Antheraea mylitta using ammonium sulphate precipitation, glycogen complex precipitation and gel filtration chromatography. Specific activity increased from 0.673 AU\\/mg in the crude digestive juice to 94.80 AU\\/mg in the final purified sample. Activity of the purified enzyme was 15-fold less than that of the digestive amylase of silkworm. Bombyx mori.

J. Nagaraju; E. G. Abraham

1995-01-01

392

Isolation and identification of a new fungal strain for amylase biosynthesis.  

PubMed

Fungi are well known for their ability to excrete enzymes into the environment. The fungal isolate FSS60 was the best amylase producer among one hundred and thirty-six isolates obtained from Syrian soils and tested for amylase production. According to the sequence of the internal transcribed spacer (ITS) rDNA gene, the isolate was identified as Aspergillus flavus. Optimal initial pH for amylase production was found to be 9.0. The enzyme was optimally active at 50 degrees C and pH 5.0. PMID:19899621

Bakri, Yasser; Magali, Masson; Thonart, Philippe

2009-01-01

393

Beta-amylase in germinating millet seeds.  

PubMed

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

Yamasaki, Yoshiki

2003-11-01

394

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

ClinicalTrials.gov

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

2013-09-04

395

Extracellular production of beta-amylase by a halophilic isolate, Halobacillus sp. LY9.  

PubMed

A moderately halophilic strain LY9 with high amylolytic activity was isolated from soil sample obtained from Yuncheng, China. Biochemical and physiological characterization along with 16S rRNA sequence analysis placed the isolate in the genus Halobacillus. Amylase production started from the post-exponential phase of bacterial growth and reached a maximum level during the early-stationary phase. The isolate LY9 was found to secrete the amylase, the production of which depended on the salinity of the growth medium. Maximum amylase production was observed in the presence of 10% KCl or 10% NaCl. Maltose was the main product of soluble starch hydrolysis, indicating a ?-amylase activity. The enzyme showed optimal activity at 60°C, pH 8.0, and 10-12.5% of NaCl. It was highly active over broad temperature (50-70°C), NaCl concentration (5.0-20.0%), and pH (4.0-12.0) ranges, indicating its thermoactive and alkali-stable nature. However, activity dropped off dramatically at low NaCl concentrations, showing the amylase was halophilic. Ca(2+) was found to stimulate the ?-amylase activity, whereas ethylenediaminetetraacetic acid (EDTA), phenylarsine oxide (PAO), and diethyl pyrocarbonate (DEPC) strongly inhibited the enzyme, indicating it probably was a metalloenzyme with cysteine and histidine resid