Salivary Alpha-amylase Activity Research Articles

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

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 increases the release of salivary alpha-amylase. Although some studies have evaluated salivary alpha-amylase under psychosocial stress, no studies have demonstrated the change in the salivary alpha-amylase (sAA) activity level in schizophrenic patients. We examined the relationship between sAA level and psychiatric state in patients with schizophrenia ( n = 54) using a portable and rapid hand-held monitor to investigate sAA. The sAA activity in the patients was significantly higher than that in the control subjects ( n = 55) ( p < 0.01). The correlation between amylase level and psychiatric symptoms was highly significant ( r = 0.37, p < 0.01). These findings indicate that higher increases in sAA may indicate severe psychiatric symptoms. These results indicate a predominant role of the sympathetic nervous system in the secretion of sAA, together with parasympathetic withdrawal, under psychosocial stress.

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

The aim of this study was to examine cardiovascular [heart rate variability (HRV)] and autonomic nervous system activation (by evaluating salivary alpha-amylase activity) that occur in professors both to, and after, the delivery of a lecture to 200 students and to determine whether gender is an influencing factor upon response. Fifty-two participants (26 women and 26 men) collected eight unstimulated saliva samples on 2 days (one a working day on which the lecture was given, the other a non-work or rest day). They also completed the Trait version of the State-Trait Anxiety Inventory (STAI) to assess their dispositional anxiety on the rest day and the State section of the STAI 15 min before and 10 min after their lecture, repeated at the same hour on the control (rest) day. The Perceived Stress Scale (PSS) was also recorded 15 min before the lecture. Continuous RR intervals were recorded before and after the lecture and the following HRV parameters were calculated: total spectral power (P (TOT)); the spectral power of the low frequency component (P (LF)); the high frequency component (P (HF)); and the ratio LF/HF. A reduction (P < 0.05) in the HF and HFnu component of HRV and an increase in the LH/HF ratio (P < 0.05) were observed at the end of the lecture. AA activity measured on the teaching day was significantly higher than that noted on the resting day. Lecturing resulted in a significant increase in the secretion of the stress marker alpha-amylase. Men and women did not differ in trait and state anxiety and no gender differences for HRV or AA activity were found.

Maternal Correlates of Children’s Stress Functioning Following a Major Natural Disaster

Concordant assessments of psychological functioning, salivary alpha-amylase (sAA), and cortisol activity were taken for children (n = 28, 6–10 yrs) and their mothers two months after being displaced by Hurricane Katrina and living in a relocation camp. Multilevel regression models revealed that the psychological functioning of the displaced children did not differ from that of demographically matched controls (n = 19 children), but the displaced group had higher sAA activity and lower cortisol levels. Compared to control mothers, displaced mothers showed higher sAA activity; greater symptoms of depression, distress, and anxiety; and lower self-esteem. Maternal psychological functioning was related to children’s endocrine activity. The results suggest that pervasive stress exposure may be associated with unique psychobiological distress regulation for mother–child dyads.

Propranolol selectively blocks the enhanced parietal old/new effect during long-term recollection of unpleasant pictures: A high density ERP study

Evidence from both animal and human research suggests that the formation of emotional memories is triggered by the beta-adrenergic system. To confirm whether modulation of central beta-adrenergic transmission is specifically involved in the neural signature of memory performance, the pre-encoding effect of propranolol (80 mg) on event-related potentials (ERPs) was measured in a placebo-controlled, double-blind, parallel-group study in 46 male healthy subjects using high density EEG and source imaging analysis during encoding and retrieval (after 1 week) of IAPS pictures of unpleasant, neutral and pleasant contents; for recognition 90 old pictures were randomly mixed with 90 new pictures. During retrieval correctly remembered old pictures elicited a significantly larger positive voltage change over the centro-parietal cortex than new pictures. Propranolol significantly reduced this old/new difference of the mean ERP amplitudes (500–800 ms) for unpleasant but not for neutral and pleasant memories. This effect correlated with salivary alpha-amylase activity, a surrogate for central adrenergic stimulation. In conclusion, propranolol selectively blocked the neural signature of unpleasant memories by mechanisms in which the parietal cortex seems to be specifically involved.

Effects of hyperoxic inhalation on psychological stress-induced salivary biomarkers

This study examined the effects of hyperoxic inhalation on psychological stress-induced salivary biomarkers. To induce psychological stress, eight males (22-24 year old) were performed a simple mathematical calculation. After the task, the subjects inspired either normal air or 100% O(2) for 30 min. The control subjects (control trial) did not perform the calculation task and inspired normal air. These three trials were randomly performed at an interval of at least one week, and the two calculation trials with and without 100% O(2) inhalation were performed using a single-blinded design. A tendency for increase in salivary cortisol (s-cortisol) and chromogranin A (s-CgA) concentrations, and a significant increase in salivary alpha-amylase (s-amylase) activity were observed following the task. Hyperoxic inhalation did not affect s-cortisol and s-CgA secretion, but decreased the s-amylase activity. Changes in the increased rate of s-amylase activity and s-CgA concentration showed a significant negative correlation with each other, after the task. These results imply that hyperoxic inhalation attenuates a part of autonomic excitability resulting from psychological stress. Although both s-amylase and s-CgA are employed as biomarkers of autonomic excitability, the s-amylase and s-CgA do not appear to be regulated by the same autonomic nervous system.

Salivary Alpha Amylase Levels under Conditions of Extreme Examination Stress

11 healthy adult graduate students were studied on two separate occasions 2 wk. apart at the same time of day. Self-rating of perceived stress and serial measurements of salivary alpha-amylase levels were performed at T1 = 15 min. before, T2 = 1 min. before, T3 = 1 min. after, and T4 = 15 min. after an event. The events were a highly stressful master's degree oral examination on the first occasion and 15 min. of typing on a computer on the second occasion as a control activity. A kinetic assay was used to estimate salivary alpha-amylase activity. Students rated higher stress on the oral examination day, and their salivary alpha-amylase activity was higher during the oral examination period than on the second less stressful control day.

Flavonoids for Controlling Starch Digestion: Structural Requirements for Inhibiting Human α-Amylase

In this study we investigated the structural requirements for inhibition of human salivary alpha-amylase by flavonoids. Four flavonols and three flavones, out of the 19 flavonoids tested, exhibited IC50 values less than 100 microM against human salivary alpha-amylase activity. Structure-activity relationships of these inhibitors by computational ligand docking showed that the inhibitory activity of flavonols and flavones depends on (i) hydrogen bonds between the hydroxyl groups of the polyphenol ligands and the catalytic residues of the binding site and (ii) formation of a conjugated pi-system that stabilizes the interaction with the active site. Our findings show that certain naturally occurring flavonoids act as inhibitors of human alpha-amylase, which makes them promising candidates for controlling the digestion of starch and postprandial glycemia.

Psychosocial stress induces working memory impairments in an n-back paradigm

In contrast to the substantial number of studies investigating the effects of stress on declarative memory, effects of stress on working memory have received less attention. We compared working memory (numerical n-back task with single digits) in 40 men exposed either to psychosocial stress (Trier Social Stress Test (TSST)) or a control condition. Task difficulty was varied using two conditions (2-back vs. 3-back). Salivary cortisol (as a marker of hypothalamus-pituitary-adrenal (HPA) activity) and salivary alpha-amylase (sAA as a marker of sympathetic nervous system (SNS) activity) were assessed immediately before and three times after the stress or control condition. As expected stress resulted in an increase in cortisol, sAA, and negative affect. Subjects exposed to stress showed significant working memory impairments in both workload conditions. The analysis of variance indicated a main effect of stress for reaction time as well as accuracy. In addition, for reaction time a stress-block interaction occurred. Follow up tests revealed that only during the first block at each level of difficulty performance was significantly impaired by stress. Thus, the effects of stress became smaller the longer the task was performed. Results provide further evidence for impaired working memory after acute stress and illustrate the time course of this phenomenon.

Neuroendocrine stress responses to an oral academic examination: No strong influence of sex, repeated participation and personality traits

Public speaking tasks have been widely used as laboratory stressors in human research. Fewer studies have investigated similar real life situations like oral examinations and results have been inconsistent. The present study investigated salivary cortisol (as a marker of hypothalamus–pituitary–adrenal (HPA) activity) and salivary alpha-amylase (sAA as a marker of sympathetic nervous system (SNS) activity) within the context of a university examination.Subjects were 40 undergraduate students who participated in an oral examination. Of these, 20 also participated in a second examination within a few weeks. Cortisol and sAA were measured immediately before and after the examination and on a control day. Additionally, subjects filled out personality questionnaires.A strong anticipatory increase in salivary cortisol and sAA as well as more modest further increases between the pre- and post-measurements were detected during the examination. Sex or oral contraceptive use had no influence on either measure. In addition, no significant differences between the first and second examination were observed.The findings indicate the neuroendocrine stress responses to laboratory stressors and to heralded real life stressors as well as the modulatory variables involved differ from each other.

Determinants of the diurnal course of salivary alpha-amylase

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 amylase and its determinants. Saliva samples were collected immediately after waking-up, 30 and 60 min later, and each full hour between 0900 and 2000 h by 76 healthy volunteers (44 women, 32 men). Compliance was controlled by electronic monitors. In order to control factors which might influence the diurnal profile of salivary alpha-amylase (such as momentary stress, mood, food, or body activity), at each sampling time point the subjects filled out a diary examining the activities they had carried out during the previous hour. Salivary alpha-amylase activity shows a distinct diurnal profile pattern with a pronounced decrease within 60 min after awakening and a steady increase of activity during the course of the day. Mixed models showed a relative independence of diurnal salivary alpha-amylase from momentary stress and other factors, but significant associations with chronic stress and mood. Our results suggest that diurnal profiles of salivary alpha-amylase are relatively robust against momentary influences and therefore may prove useful in the assessment of sympathetic nervous system activity. The findings underscore the need to control for time of day in studies using salivary alpha-amylase as a dependent variable.

Flat-Chip Microanalytical Enzyme Sensor for Salivary Amylase Activity

It is considered that measurement of salivary alpha-amylase activity is a useful tool for evaluating the sympathetic nervous system. The purpose of this research is to demonstrate a new design of a flat-chip microanalytical enzyme sensor (flat-chip sensor) for salivary amylase activity as a Micro-Electro-Mechanical Systems (MEMS), which may be used for wearable analytical systems. To meet this purpose, the biosensor needs to be miniaturized and to possess high-sensitivity. A pre-column and a flat-enzyme electrode were incorporated in a flow cell of volume 25.7 ml. In order to miniaturize the flow cell, two enzymatic membranes containing maltose phosphorylase obtained from Enterococcus hirae (MP membrane) and glucose oxidase and peroxidase (GOD-POD membrane) were immobilised on the same planar surface. As a result, a flat-chip sensor incorporating a flow cell as small as a C battery was produced. The optimum conditions of three parameters of the fabricated flat-chip sensor, the immobilising method of the enzymatic membrane, dropping volume of the mixed enzymatic solution and flow rate of the sample solution, were examined. An analytical system for 0-190 kU/l amylase activity with R(2) of 0.97 was fabricated with a sample volume of 50 microl. This research indicates the possibility of a wearable biosensor for continuous monitoring of salivary amylase activity.

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

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.

Human salivary alpha-amylase Trp58 situated at subsite -2 is critical for enzyme activity.

The nonreducing end of the substrate-binding site of human salivary alpha-amylase contains two residues Trp58 and Trp59, which belong to beta2-alpha2 loop of the catalytic (beta/alpha)(8) barrel. While Trp59 stacks onto the substrate, the exact role of Trp58 is unknown. To investigate its role in enzyme activity the residue Trp58 was mutated to Ala, Leu or Tyr. Kinetic analysis of the wild-type and mutant enzymes was carried out with starch and oligosaccharides as substrates. All three mutants exhibited a reduction in specific activity (150-180-fold lower than the wild type) with starch as substrate. With oligosaccharides as substrates, a reduction in k(cat), an increase in K(m) and distinct differences in the cleavage pattern were observed for the mutants W58A and W58L compared with the wild type. Glucose was the smallest product generated by these two mutants in the hydrolysis oligosaccharides; in contrast, wild-type enzyme generated maltose as the smallest product. The production of glucose by W58L was confirmed from both reducing and nonreducing ends of CNP-labeled oligosaccharide substrates. The mutant W58L exhibited lower binding affinity at subsites -2, -3 and +2 and showed an increase in transglycosylation activity compared with the wild type. The lowered affinity at subsites -2 and -3 due to the mutation was also inferred from the electron density at these subsites in the structure of W58A in complex with acarbose-derived pseudooligosaccharide. Collectively, these results suggest that the residue Trp58 plays a critical role in substrate binding and hydrolytic activity of human salivary alpha-amylase.

Characterization of the α-amylase Receptor of Streptococcus gordonii NCTC 7868

The purpose of the work described here was to investigate the mechanisms involved in the binding of salivary alpha-amylase to Streptococcus gordonii NCTC 7868 (Challis). Of six types of alpha-amylase studied, only mammalian forms of the enzyme were found to bind to S. gordonii cells. Salivary alpha-amylase binding was inhibited by treatment of cells with trypsin and pronase, but not with pepsin or sodium periodate. Presence of starch, dextrin, or maltoheptaose partially inhibited binding of the enzyme to S. gordonii. Both mutanolysin extracts of cells and culture supernatants contained alpha-amylase-binding activity, which was partially purified by Sepharose CL-6B and DEAE-ion-exchange chromatography. Western blotting detected four putative receptor bands--65 kDa, 15 kDa, 12.5 kDa, and one with a very high molecular weight; the lower-molecular-weight components may be products of proteolytic degradation of the high-molecular-weight material, but their true relationship has yet to be determined. Pre-treatment of salivary alpha-amylase with these putative receptors partially inhibited subsequent binding of the enzyme to S. gordonii cells. When bound to cells, only 19% of the salivary alpha-amylase activity was detectable, suggesting that alpha-amylase binds to the receptor at or near the active site of the enzyme.

Sialochemische Untersuchungen beim Morbus Sjögren

The concentrations of sodium, potassium, calcium, protein and lactoferrin and the enzymatic activity of salivary kallikrein, phosphohexoisomerase, alpha-amylase, lysozyme and IgA were measured in the parotid saliva of 12 patients suffering from Sjögren's syndrome. Compared with a control group (n = 31) remarkable differences were found in the composition of the parotid saliva in Sjögren patients. The possible reasons for these sialochemical alterations are discussed.

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

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

Preparation of human salivary alpha-amylase specific monoclonal antibody.

A mouse hybridoma cell line which produced an anti-human salivary alpha-amylase monoclonal antibody was obtained by fusion between mouse spleen cells immunized with human salivary alpha-amylase and mouse myeloma cells, followed by screening the hybridoma cells by enzyme-linked immunosorbent assay. The hybridoma cell line (27-4-1) secreted IgG. The monoclonal antibody produced by the hybridoma showed no inhibitory effect on the activity of human salivary alpha-amylase. The specificity and reactivity of this monoclonal antibody were examined by determining the activities of human salivary and pancreatic alpha-amylases bound to the monoclonal antibody immobilized on polystyrene balls or by enzyme immunoassay with the monoclonal antibody conjugated with beta-D-galactosidase. The results revealed that the monoclonal antibody produced by the hybridoma cell line was specific for salivary alpha-amylase and absolutely unreactive to pancreatic alpha-amylase.

Amylase inhibitor from wheat: its action and clinical application.

The electrophoretic and column chromatographic characteristics of an amylase inhibitor of wheat origin were investigated. Further, the clinical usefulness of this inhibitor for determining the ratio of pancreatic to salivary isoamylase activity in serum was evaluated. Amylase inhibitor inhibits the action of salivary alpha-amylase by making an amylase-inhibitor complex, which is easily separated into its individual component during electrophoresis with full recovery of amylase activity. Using the specific inhibitory effect of this inhibitor on salivary alpha-amylase activity, the ratio of pancreatic to salivary isoamylase activity (P/S) in serum was determined. There was a good correlation in P/S ratio in serum between the results obtained with the inhibitor method and those with electrophoretic method. The P/S ratio in sera from patients with acute pancreatitis was over 8.0, whereas that in sera from patients with salivary-type hyperamylasemia such as mumps, pulmonary diseases and following surgery was less than 0.1. However, hyperamylasemia due to macroamylase or renal failure could not be identified by the inhibitor method.