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Background: A reliable, sensitive and practicable method for the measurement of intestinal lactase activity is needed., Method: The assay is based on the continuous measurement of released glucose by a coupled hexokinase/glucose-6-phosphate dehydrogenase (HK/G6PDH) reagent., Results: The procedure was first optimized for lactase from rat intestinal mucosa. The optimum pH is 6.5 and apparent Km was 17 mmol/l for lactose. The procedure was also adapted on a Cobas Mira automated analyzer; progress curves of the reaction rate can be continuously monitored. The automated assay correlated strongly with the conventional method of Dahlqvist (r(2) = 0.996). The described method has also been applied to human intestinal mucosa biopsies after determination of the catalytic properties of human enzyme (optimum pH 6.0 and apparent Km 34 mmol/l)., Conclusion: The HK/G6PDH method is reliable, rapid, sensitive and easy to perform both manually as well as in the automated version. It is optimized for human and rat intestinal lactase., (Copyright 2003 S. Karger AG, Basel)

A new method for the assay of maltase and sucrase is reported. The method makes use of mutarotase, hexokinase and glucose 6-phosphate dehydrogenase as ancillary enzymes. The reaction is linear at least up to a delta E/min of 0.13.

A collection of wild-type and reference strains of Streptococcus mutans and Streptococcus sobrinus was compared using a set of fermentation and enzymic tests in order to establish a simple scheme for the differentiation of these two species isolated from selective media. Tests for the production of acid from N-acetylglucosamine, arbutin and melibiose, and the presence of alpha-galactosidase and alpha-glucosidase activities were useful in differentiating these species. Strains of Streptococcus mutans unable to ferment melibiose were also frequently unable to ferment other sugars; a consideration of the genetic control and organization of the genes involved in melibiose metabolism indicates that certain of the conventional characterization tests are not measurements of independent biochemical capabilities.

Rat jejunal disaccharidases were measured after feeding high fat fibre-free, high fat pectin, high fat cellulose, high fat galactomannan and high fat tannin diets for 27 weeks. Compared with the high fat fibre-free diet, pectin lowered both lactase and sucrase levels (p less than 0.025). Galactomannan and tannin lowered only the lactase level (p less than 0.05), while cellulose had no significant effect on disaccharidase levels. These data suggest that dietary fibre components have individual and specific effects on jejunal disaccharidase levels, in the presence of a high fat diet.

Previous work in our laboratory and in others suggest that protein malnutrition plays an important role in the pathogenesis of hepatic dysfunction after jejunoileal bypass for morbid obesity. This experimental study was undertaken to attempt to correlate hepatic dysfunction (the criterion used was the bromsulphalein clearance) to morphological and enzymatic adaptation of the functioning intestine in the rat. It was observed that the period of impaired bromsulphalein clearance is concomitant with a slight ileal morphological adaptation and especially with a period of selective adaptation of maltase and sucrase activities, whereas there is no increase in aminopeptidase activity. These data support the hypothesis that after jejunoileal bypass a preferential absorption of carbohydrates along with a protein deficiency state could occur and as in kwashiorkor it results in an impaired nutritional status, a major contributing factor to bypass-induced liver injury.

Wistar rat jejunal disaccharidases were measured after feeding low fat, low fat + 5% pectin, low fat + 0.4% galactomannan, high fat, high fat + 5% pectin and high fat + 0.4% galactomannan diets for 16 weeks. All rats fed high fat diet had significantly lower jejunal sucrase and maltase levels when compared with their respective low fat groups. Lactase was significantly lowered in the high fat pectin group compared with the low fat pectin group, but was not significantly different when comparing the high fat or high fat galactomannan with their respective low fat groups. There was no significant difference in lactase, sucrase or maltase levels between the low fat groups, or between the high fat groups. We conclude that in Wistar rats an increased dietary fat level lowers jejunal sucrase, maltase and lactase levels, while the gel-forming substances pectin and galactomannan added at the levels of 5% and 0.4%, respectively, have no effect.

Activities of seminal plasma alpha-D-glucosidase, beta-D-galactosidase, alpha-amylase, chymotrypsin-like enzyme and RNAase were studied in normo spermic, oligospermic and non-obstructive azoospermic men. No significant change in the activities of the aforesaid enzymes were found amongst the three catagories, classified according to sperm density.