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

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.     

Preventing baker's asthma: an alternative strategy

BACKGROUND: Although baker's asthma has attracted considerable research interest over the last 30 years, success in its prevention has been slow to achieve. This paper describes the results of an alternative preventive strategy, based on an observation that the excess of sensitization in bread bakers is largely due to IgE-mediated allergy to fungal amylase, contained in bread improvers. The practical application of the strategy has been to limit bread improver exposures to <1 mg/m3 [8 h time-weighted average (TWA)], whilst exposures to all other ingredients, including flour, have been limited to <10 mg/m3 (8 h TWA). METHOD: The paper describes the findings of in-house respiratory health surveillance and dust sampling programmes, from a UK food company whose primary interests are milling and baking, over the period following the introduction of the strategy to target the reduction in bread improver exposure. RESULTS: Over the 10 year period of surveillance, the incidence of symptomatic sensitization in the bread baking sector (2240 per million employees per annum) was greater than for the other flour-using groups (330 per million employees per annum), despite broadly similar total inhalable dust exposures. There was an overall reduction in the incidence of new cases of symptomatic sensitization, from 2085 per million employees per year in the first 5 years of the surveillance programme, to 405 per million employees per year in the subsequent 5 years. CONCLUSION: The strategy of targeting bread improver exposure is an effective approach for the prevention of new cases of symptomatic sensitization in bread bakeries.     

Acute pancreatitis coincident with valproate use: a critical review

PURPOSE: Acute pancreatitis has been associated with a number of medications, including valproate (VPA). Valproate-coincident pancreatitis is uncommon and is usually associated with other risk factors; however, the United States Food and Drug Administration has issued a box warning for risk of acute pancreatitis with VPA products. We reviewed cases of pancreatitis in VPA-treated patients from the clinical database of VPA/divalproex trials and compared the incidence of elevated amylase levels between VPA- and placebo-treated patients. METHODS: Report rates of acute pancreatitis coincident with VPA use in 34 clinical trials were calculated. Incidence rates of amylase elevations above the normal range in three clinical placebo-controlled migraine trials were compared between VPA- and placebo-treated patients. RESULTS: Among 3,007 VPA-treated patients in 34 clinical trials, two reports of pancreatitis were considered by investigators to be probably related to VPA. Both patients recovered. Similar rates of amylase elevations were observed in VPA- (5.9%) and placebo-treated (6.1%) patients in the three migraine headache trials. CONCLUSIONS: VPA-coincident acute pancreatitis is uncommon and idiosyncratic. Checking amylase levels in the absence of other clinical signs and symptoms provides little value for predicting pancreatitis. Physicians should be guided by clinical symptoms of pancreatitis to identify cases.     

High-calorie, rice-derived, short-chain, glucose polymer-based oral rehydration solution in acute watery diarrhea

In this study, we have compared the effects of the World Health Organization oral rehydration solution (WHO ORS) and an ORS containing short polymers of glucose (Amylyte ORS) at a high caloric density (five times) and comparable osmolality, on stool output, duration of diarrhea, weight gain and fluid and electrolyte balance, in randomized, open-labeled, controlled clinical trials in five centers. A total of 198 male children (4 months to 10 years) with acute diarrhea (<72 h after onset) were assigned by random allocation to either WHO ORS or Amylyte ORS at five centers in Asia. Children were stratified according to grade of dehydration (mild, moderate or severe) and the initial purging rates during the first 6h (low (<2ml/kg/h), moderate (2–5ml/kg/h) and high (>5ml/kg/h) purgers). The clinical characteristics of the children in the two treatment groups were comparable. Amylyte ORS reduced stool volumes significantly in children with severe dehydration (285.4 ± 74.2 versus 75.5 ±20.0 ml/kg; p < 0.05) and in children with a high initial purging rate (200.3 ±42.8 versus 130.5 ± 9.1 ml/kg; p < 0.05). This was accompanied by a significant (276.4 ±14.6 versus 227.6 ± 11.8 ml/kg; p < 0.01) reduction in ORS requirements in the Amylyte ORS treated group, the effect being greatest in children with severe dehydration (491.5 ± 108.5 versus 155.7 ± 27.3 ml/kg; p < 0.01) or high initial purging rates (394.2 ± 66.2 versus 316.8 ± 34.8 ml/kg; p < 0.05). In mild or moderate dehydration and low or moderate purgers, duration of diarrhea in children treated with Amylyte ORS was not reduced. A significant (p < 0.05) reduction in duration of diarrhea was found in children with severe dehydration (56.4 ± 1.8 versus 34.0 ± 5.6h) or high purging rates (51.5 ± 7.1 versus 38.2 ± 4.8 h). Significantly (p < 0.0001) greater weight gain (520.3 ± 48.5 g versus 228.5 ± 42.1 g) and percent gain in weight (4.8 ± 0.4% versus 2.3 ± 0.4%) were found in children given Amylyte ORS compared with those given WHO ORS. Amylyte ORS rehydrated children with acute diarrhea, reduced stool volume, duration of diarrhea and ORS requirements in children with severe dehydration or high initial purging rates, and improved weight gain. Amylyte ORS has the advantage of containing short polymers of glucose that provide over five times the calories per liter than the glucose-based WHO ORS.     

Ca2+ dependence of the release of noradrenaline during nerve degeneration in the parotid gland

Summary Amylase secretion in vitro was used as an indication of the degeneration activity of sympathetically denervated parotid glands. Seventeen and a half hours after sympathetic denervation, slices of parotid gland released amylase into the incubation medium at a rate higher than that observed for non-denervated glands. The time course of amylase release from denervated glands followed a bellshaped pattern similar to that observed in other denervated structures. Lowering extracellular Ca²⁺ to 0.25 mmol/l diminished significantly the release of amylase. Low Ca²⁺ however, did not decrease the amylase release in response to added noradrenaline.These results indicate that Ca²⁺ is required for the release of noradrenaline from degenerating nerve endings.     

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

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 done to look at the way these systems interact, especially when both systems are activated. Additionally, sex effects in CORT responses have been investigated relatively often, but possible sex differences in sAA levels and responses, or the way both systems interact has not been the focus of sufficient studies to yield a univocal conclusion. In this study we presented a group of healthy participants (n=80) with two mildly stressful tasks, consisting of an aversive picture rating task and a cold pressor stress (CPS) task. The second task was compared with a control task. We expected a rise in sAA level in response to the first task and sAA as well as CORT responses on the second task and explored the interaction between the two responses. Results indicate that sAA is indeed a sensitive marker in both psychologically and physically induced arousal paradigms, whereas a cortisol response was only observed in the CPS task. Men had higher sAA levels than women during the complete course of the study, but men and women were comparable in their responsivity to the tasks. No strong correlations between sAA and CORT responses were found.     

Immunoelectron microscopy of amylase in the human parotid gland

Summary Specimens of the human parotid gland were studied by immuno-electron microscopy for the presence of amylase. Both the protein A-gold technique and the biotin-avidin-gold technique were used on the same specimens. Different fixations were tried. Amylase was detected in the zymogen granules in high amounts. This enzyme could even be seen in glutaraldehyde fixed and routinely embedded material. The subcellular localization of this enzyme opens a new field of functional morphological studies and studies in special tumours including acinic cell carcinomas.Supported by the Deutsche Forschungsgemeinschaft and by the Hamburger Stiftung zur Förderung der KrebsbekämpfungDedicated to Prof Dr. med. Dres. h.c. Wilhelm Doerr, Heidelberg, in honour of his 70th birthday     

Decreased pancreatic CCK receptor binding and CCK-stimulated amylase release in Zucker obese rats

In Zucker obese rats the response to the effects of CCK on food intake and pancreatic exocrine function are decreased. However, it is unknown whether the decreased responsiveness is due to decreased receptor number and/or sensitivity or abnormal circulating concentrations of CCK. In these experiments percent total binding of 125I-CCK-33 to pancreatic acini from obese rats was one-half that in lean rats when data was expressed on a per microgram DNA basis (19.6 +/- 5.1 vs. 47.4 +/- 11.4, p less than 0.01). In a second experiment while the maximally effective dose of CCK for stimulating amylase secretion from dispersed pancreatic acini was similar in obese and lean rats (10(-10) M), less amylase was secreted in obese rats across the dose range tested (p less than 0.001). In contrast, carbachol had similar potency and efficacy in stimulating amylase release from obese and lean pancreatic acini. The increase of pancreas size by use of a trypsin inhibitor was greater in lean than obese rats (p less than 0.03). In addition, stimulation of amylase release by CCK from obese trypsin inhibitor-treated compared with control obese rats was greater than that from lean trypsin inhibitor-treated compared with control lean rats (p less than 0.002). However, overall, stimulation of amylase secretion by CCK was only 36% of control (p less than 0.001) and by carbachol was only 20% of control (p less than 0.001). Thus, increased size by increased cell number was associated with decreased response per cell.(ABSTRACT TRUNCATED AT 250 WORDS)     

Dynamics of beta adrenoceptor induced amylase release and cyclic AMP accumulation in the guinea pig submandibular gland

The dynamics of amylase release from the guinea pig submandibular gland were studied in vitro by applying a multi-channel microperifusion set. This technique makes it possible to measure time related enzyme release more accurately and to take samples of perifused tissue at short intervals. Stimulation of the ^-adrenoceptor with norepinephrine gives rise to a rapid initial enzyme discharge, detectable within 15 s. Administration of propranolol inhibits enzyme release, which is not restored after removal of the agent. Simultaneous measurements of tissue cyclic AMP during norepinephrine stimulation at various time intervals display a significant increase of cAMP as early as 15 s after stimulation of secretion. This increase of cAMP thus coincides with the discharge of amylase. In addition, cAMP continuously accumulates during 30 min of norepinephrine perifusion of the slices. The present study describes a valuable tool with high sensitivity for visualizing the relations between enzyme secretion from the salivary gland and the intracellular biochemical processes. The data obtained further indicate a close correlation between amylase and cAMP during the initial phase of enzyme discharge.