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Circadian rhythms in disaccharidases of rat small intestine and its relation to food intake.

The activities of maltase and sucrase of the small intestine were low at night and high in the daytime in rats which had been fed from 09.00 h to 15.00 h for 2 weeks. A remarkable rise of enzyme activities was observed at 08.00 h, 1 h before the start of feeding. The rhythmic changes in disaccharidase activities continued for at least 2 days after starvation, but completely disappeared after 5 days of starvation. It was suggested that the disaccharidase rhythms are not a direct consequence of food intake, but that anticipation of food intake acts as a trigger for initiation of the disaccharidase rhythms.

Circadian Rhythm↗

Intestinal brush border membrane-bound disaccharidases of the American alligator, Alligator mississippiensis.

1. The disaccharidase activities of the small intestines of American alligators (Alligator mississippiensis) were studied in epithelial scrapes and brush-border membrane preparations. 2. Maltase, isomaltase and trehalase activities were found. Activities of these enzymes were higher in the proximal small intestine and decreased distally. 3. Disaccharidase activities were enriched 12-15 times in brush-border membrane preparations, compared with mucosa/enterocyte crude homogenates and were co-enriched with the brush-border membrane marker alkaline phosphatase. 3. The pH optima were: maltase 6.5; isomaltase 5.6; and trehalase 5.8. The Q10 of maltase, the most active enzyme, was equal to 1.82. 4. In reptiles, as in mammals, disaccharidase activities may be correlated with feeding habits. The co-occurrence of sucrase and isomaltase may not be a common feature of vertebrates.

Alligators and Crocodiles↗

Intestinal disaccharidases and some renal enzymes in streptozotocin-induced diabetic rats fed sapogenin extract from bitter yam (Dioscorea polygonoides).

In this study, the effects of bitter yam sapogenin extract or commercial diosgenin on intestinal disaccharidases and some renal enzymes in diabetic rats were investigated. Diabetic male Wistar rats were fed diets supplemented with 1% sapogenin extract or commercial diosgenin for 3 weeks. Plasma glucose, intestinal disaccharidases and the activities of transaminases, acid phosphatase, glucose-6-phosphatase, ATP citrate lyase, glucose-6-phosphate dehydrogenase and pyruvate kinase were assessed for the level of metabolic changes in the kidney of diabetic rats. Sapogenin extract or commercial diosgenin supplementation resulted in a significant decrease in lactase and maltase activities in all three regions of the intestine compared to the diabetic control group. However, the test diets significantly reduced intestinal sucrase activity in the proximal and mid regions. Test diets supplementation resulted in a significant decrease in the activities of the transaminases compared to the normal and diabetic control groups. The activity of glucose-6-phosphatase was significantly increased while the activities of ATP citrate lyase, pyruvate kinase and glucose-6-phosphate dehydrogenase were significantly reduced in the kidney of the diabetic control rats compared to the normal group. Test diets supplementation did not significantly alter glucose-6-phosphatase, ATP citrate lyase and pyruvate kinase activities compared to the diabetic control. However, there was a significant increase in glucose-6-phosphate dehydrogenase activity toward the normal group. In conclusion, the consumption of bitter yam sapogenin extract or commercial diosgenin demonstrated hypoglycemic properties, which are beneficial in diabetes by reducing intestinal disaccharidases activities; however, bitter yam sapogenin extract may adversely affect the integrity of kidney membrane.

Animals↗

Comparison of endoscopic forceps biopsies and capsule biopsies in determining disaccharidase activity in the duodenum.

Endoscopic biopsy is an accepted technique for obtaining samples of small bowel mucosa for histological examination. In this study a comparison between endoscopic forceps biopsies and capsule biopsies in estimating disaccharidase enzyme activity in the small bowel was evaluated. Disaccharidase levels on 168 consecutive small bowel biopsies obtained by either endoscopic biopsy or capsule biopsy were reviewed retrospectively. There was no significant difference in enzyme activity in samples collected by either method. Another 18 patients had simultaneous endoscopic and capsule biopsies from the second part of the duodenum. Again, there was no difference in enzyme activity between samples collected by either technique at the same level in the duodenum. The disaccharidase activity in forceps biopsies taken from the second part of the duodenum is equivalent to those obtained by capsule biopsy.

Adult↗

Disaccharidase activities in small intestinal mucosa in patients with cystic fibrosis.

The disaccharidase activities in small intestinal biopsies were related to the morphology of the mucosa and the ages of 63 patients with cystic fibrosis and 177 healthy control subjects of Caucasian origin. In patients with CF and in the healthy control subjects under 5 years of age with normal intestinal mucosa, no low lactase activity was found. In those patients with CF who were over 5 years of age, one group had high and one group had low lactase activity, as occurs in healthy Caucasian control subjects of the same age. This finding supports the view that in patients with CF, lactase deficiency is not related to the disease entity. In patients with or without CF who had the same degree of mucosal atrophy, the decrease of disaccharidase activities followed the same pattern, indicating that enzyme activities are affected to the same extent by the damage of the mucosa. In patients with CF with pancreatic insufficiency, the disaccharidase activities were significantly (P less than 0.001) higher when compared to those in control subjects of the same age and ethnic group, although the increase was not uniform in all patients with cystic fibrosis.

Atrophy↗

Purification of a fraction of Giardia lamblia trophozoite extract associated with disaccharidase deficiencies in immune Mongolian gerbils (Meriones unguiculatus).

The effects on disaccharidase activities of challenging gerbils previously exposed to Giardia lamblia with fradions of the crude trophozoite extract were examined. Gel filtration of the soluble extract on a Sephacryl S-200 HR column resulted in 3 fradions: F1, F2 and F3. Only a challenge with fraction F1 (0.1 mg total dose) was found to induce disaccharidase deficiencies. Boiling F1 prior to challenge did not change this effect on the enzyme activities. However, the decreases were not obtained when the total F1 dose was reduced to 0.05 mg. Column chromatography of fraction F1 under dissociating and reducing conditions resulted in 2 further fractions: F1a and F1b. Challenging immune gerbils with F1b led to impairments of disaccharidose activity similar to those obtained with F1. Protein analysis of the crude extract, as well as the fractions of the extract, revealed several high and low molecular weight bonds. These findings indicate that a constituent(s) of fraction F1b is the portion of the parasite which induces disaccharidase deficiencies in immune gerbils. This fraction consists of proteins ranging in molecular weight from 32 to 200 kDa. In addition the G. lamblia fraction involved in the decreases in enzyme activity is heat-stable.

Animals↗

Small bowel disaccharidase activity in the rat as affected by intestinal resection and pectin feeding.

This study investigated the effects of small bowel resection (SBR) and a pectin-supplemented elemental diet (ED) on intestinal disaccharidase activity. Rats underwent placement of feeding gastrostomy and swivel apparatus. Control animals were returned to their cages while resected animals underwent an 80% SBR. Postoperatively, animals received either a pectin-free ED or the ED supplemented with 2% pectin. After 2 wk jejunal and ileal mucosal sucrase, maltase, and lactase activities and protein content were determined. Feeding the ED after SBR resulted in significant increases in all three ileal segmental disaccharidase activities but only maltase activity was significantly increased in the jejunum. The pectin-supplemented ED, however, significantly enhanced the adaptation of jejunal and ileal segmental sucrase, maltase, and lactase activity to SBR with the increase in all three jejunal disaccharidase activities being significantly greater than that of the resected animals fed the ED alone.

Adaptation, Physiological↗

Effect of an intestinal disaccharidase inhibitor (AO-128) on obesity and diabetes.

A new disaccharidase inhibitor, AO-128, showed 190-3900-fold more potent inhibition of purified rat small intestine sucrase-isomaltase (S-1) complex and 23-33-fold more potent inhibition of semipurified porcine small intestine disaccharidases than acarbose. AO-128 suppressed elevation of the blood glucose concentration after oral sucrose, maltose, and starch, but not after oral glucose, fructose, and lactose. The chronic addition of AO-128 to the diet produced antiobesity and antidiabetic actions in obese and/or diabetic animals. Undesirable side effects, such as diarrhea and soft feces, were observed only for the first 5-7 d and suppression of intestinal disaccharidase activities was observed even at the end of the experiment, suggesting that the suppressive or delaying effect of AO-128 on elevation of the postprandial blood glucose concentrations is involved in reduction in body weight gain and prevention and/or amelioration of the diabetic state. Thus, AO-128 is useful as an adjunct to the dietary management of obesity and diabetes.

Acarbose↗

Response of activity of jejunal disaccharidases and pancreatic amylase in young and middle-aged rats to a high carbohydrate diet.

Sprague-Dawley rats fed a nonpurified diet from weaning to 3 mo (young) or 12 mo (middle aged) of age were fed a low (5 cal%) starch (LST) purified diet for 2 wk. They were then fed a high (70 cal%) starch (HST) purified diet for up to 4 wk. Body weights of both age groups were greater in rats fed the LST than in those fed nonpurified diet throughout. The young, but not the middle-aged, group continued this gain while consuming HST. The levels of activity of intestinal disaccharidases of upper (proximal one-third) and lower (middle one-third) jejunum, and pancreatic alpha-amylase were lower in rats fed LST diet in both age groups than in rats fed HST. Similar increases of specific (per protein or DNA) activity were observed in both age groups throughout the first three (disaccharidase) to four (pancreatic alpha-amylase) days of feeding HST. Values of specific activity of the middle-aged group returned to initial levels with continued feeding of HST. In contrast, values in the young group tended to plateau (disaccharidases) or continued to increase (alpha-amylase). Differences in adaptability over 1 mo were most dramatic for alpha-amylase and sucrase, but negligible for lactase.

Aging↗

Intestinal disaccharidase and alkaline phosphatase activity in giardiasis.

We report results on determinations of small intestinal brush-border enzyme activities in 22 children (aged 11 months to 14 years) with giardiasis. In particular, activities of disaccharidases (lactase, sucrase, maltase) and of alkaline phosphatase were investigated. Forty-one percent of the patients, irrespective of age, had a demonstrable depression of disaccharidase activities, usually in a combination involving two or more enzymes. A depression of intestinal alkaline phosphatase activity was present in 33% of patients, and only in those who demonstrated disaccharidase deficiencies. Mild villus atrophy was present in two mucosal specimens, whereas all others showed normal villus morphology by light microscopy. The results obtained in this study suggest that giardiasis in otherwise healthy children does not cause marked structural damage to the small bowel mucosa, as seen by the light microscope. However, some form of damage to the brush border does occur frequently, as evidenced by a depression of brush-border enzymes. This damage most likely contributes to the diarrhea and also to the carbohydrate intolerance in these patients.

Adolescent↗

Disaccharidase activities, jejunal morphology, and carbohydrate tolerance in children with chronic diarrhea.

Children with chronic diarrhea were examined for their carbohydrate tolerance, small bowel morphology, and specific disaccharidase activities to determine whether disaccharidase enzymatic activity is related to villus height and thus may be viewed as an independent variable that controls digestive capacity. Results indicated that 65-78% of all abnormal biopsy samples had specific disaccharidase activities that fell within the normal range. Clinical disaccharide tolerance studies indicated that specific enzyme activity did not correlate with disaccharide tolerance.

Adolescent↗

Ethnic differences in intestinal disaccharidase values in children in Finland.

BACKGROUND: Intestinal disaccharidase activities tend to be low in villous atrophy, but there are only a few reports of enzyme activities in children with normal villous architecture. METHODS: In the current study the data were reviewed on disaccharidase activities in duodenal biopsy specimens of normal villous structure in 223 children undergoing upper gastrointestinal endoscopy in 1997 and 1998. The ancestry was Finnish in 188 children (median age 8.0 years; range, 0.2-18 years), African in 27 children (median age 5.0 years; range, 1-13 years), and other in eight children. RESULTS: The mean activities of lactase, sucrase, and maltase were significantly higher in Finnish children than in children of African origin (P < 0.0001, P < 0.002, and P < 0.02, respectively). Lactase activity decreased with increasing age (P < 0.001), but age had no significant effect on maltase and sucrase activities. Among Finnish children, 31% (59/188) had lactase activity below the established reference range of 20 units (units are micromoles of substrate hydrolyzed per minute at 37 degrees C per gram of protein) and one child had a probable sucrase-isomaltase deficiency. When these 60 children with low enzyme activities were excluded, the geometric means were lactase, 35.7 units (95% confidence interval [CI], 32.8-38.6 units); maltase, 241 units (95% CI, 225-258 units); and sucrase, 57.5 units (95% CI, 53.5-61.6 units). Among the children of African origin, lactase activity was decreased in 67% (18/27). All three enzyme activities were decreased in parallel more often among the African children (8/27) than among the Finnish children (9/188; P < 0.002). CONCLUSIONS: Ethnicity has a strong effect on disaccharidase values in children with normal villous structure. African children have lower activities of lactase, sucrase, and maltase in duodenal specimens than do children of Finnish origin.

Adolescent↗

Disaccharidase activities in dyspeptic children: biochemical and molecular investigations of maltase-glucoamylase activity.

BACKGROUND: Maltase-glucoamylase enzyme plays an important role in starch digestion. Glucoamylase deficiency is reported to cause chronic diarrhea in infants, but its role in dyspeptic children is unknown. METHODS: Glucoamylase and other disaccharidase specific activities were assayed from duodenal biopsy specimens in 44 children aged 0.5-18 years (mean, 10 +/- 5 years) undergoing endoscopy to evaluate dyspeptic symptoms. All subjects had normal duodenal histology. Intestinal organ culture was used to evaluate synthesis and processing of maltase-glucoamylase. Sequencing of the maltase-glucoamylase coding region was performed in subjects with low activity or variation of isoform in organ culture. RESULTS: Twenty-two of the dyspeptic children had one or more disaccharidases with low specific activity. Twelve subjects (28%) had low activity of glucoamylase. Eight subjects had low activities of glucoamylase, sucrase, and lactase. Low glucoamylase activity was not correlated with the isoform phenotype of maltase-glucoamylase as described by metabolic labeling and sodium dodecyl sulfate electrophoresis. Novel nucleotide changes were not detected in one subject with low glucoamylase activity or in two subjects with variant isoforms of maltase-glucoamylase peptides. CONCLUSION: Twelve of 44 dyspeptic children had low specific activity of duodenal maltase-glucoamylase. Eight of these children had low specific activity of all measured disaccharidases.

Biopsy↗

Disaccharidase activities in jejunal fluid.

Jejunal fluid and mucosal tissue were obtained simultaneously from the same jejunal site in a group of 29 children by a modified biopsy procedure. Lactase, maltase, and sucrase activities were measured in both fluid and mucosal specimens using the same analytical method. The fluid enzyme activities showed highly significant positive correlations with the same enzyme activity in the relevant tissue samples. Relative concentrations of disaccharidase enzymes represented by sucrase: lactase activity ratios also showed a highly significant positive correlation between fluid and tissue. This close relation suggests that the mucosa is the sole or predominant source of disaccharidase activity in the intestinal fluid. The results of kinetic studies comparing tissue and fluid enzyme characteristics also indicate a mucosal origin for the fluid enzyme activities. We conclude that disaccharidase activities in jejunal fluid reflect closely local tissue values and that these measurements may be useful in assessing mucosal enzyme activity in infants in whom jejunal biopsy is not possible.

Adolescent↗

Microvillous surface area in secondary disaccharidase deficiency.

Electron microscopy was used to measure microvillous surface area in seven small intestinal biopsies showing secondary disaccharidase deficiency and in five biopsies having normal disaccharidase levels. There were significant reductions in microvillous surface area in the enzyme deficient group, although the microvilli were not severely damaged. Histological abnormality was not always present with disaccharidase deficiency. There was no significant difference in intraepithelial lymphocyte counts between the two groups.

Adolescent↗

Features of small intestinal pathology (epithelial cell kinetics, intraepithelial lymphocytes, disaccharidases) in a primary Giardia muris infection.

In an attempt to correlate host and parasite-related events occurring during the course of a primary Giardia infection in the mouse we have measured epithelial cell kinetics, enzymes, and intraepithelial lymphocytes at different stages of the infection. New methods were developed for the accurate measurement of parasite numbers and distribution within the gut. In jejunum a modest decrease in villus length and intraepithelial lymphocytes at week 1 preceded a pronounced disaccharidase deficiency at week 2, the time of maximum trophozoite numbers, whereas crypt lengthening and increased cell production became maximal at week 3. As trophozoite numbers fell the intraepithelial lymphocyte count and disaccharidase values rose. With the exception of the intraepithelial lymphocyte count, which followed the same pattern as in jejunum but two weeks later, the changes seen in the ileum were the opposite of those in jejunum, suggesting rapid ileal adaptation. The results indicate that the disaccharidase deficiency associated with giardiasis is likely to represent a direct effect of the parasite on the brush border rather than enterocyte immaturity, whereas the intraepithelial lymphocyte response reflects host immunity to the parasite. Profound adaptive changes occur throughout the small intestine in response to a relatively localised insult.

Animals↗

Evaluation of differential disaccharide excretion in urine for non-invasive investigation of altered intestinal disaccharidase activity caused by alpha-glucosidase inhibition, primary hypolactasia, and coeliac disease.

BACKGROUND/AIM: The reliability of a quantitative method for the non-invasive assessment of intestinal disaccharide hydrolysis was assessed. METHODS: Differential excretion of intact disaccharide, expressed as ratios of lactulose to appropriate hydrolysable disaccharides in urine collected following combined ingestion, has been investigated in healthy volunteers with drug induced alpha-glucosidase inhibition, in subjects with primary hypolactasia, and patients with coeliac disease. RESULTS: Oral administration of the alpha-glucosidase inhibitor 'Acarbose' (BAY g 5421, 200 mg) together with sucrose and lactulose increased the urinary sucrose/lactulose excretion ratios (% dose/10 h) fivefold. The effect was quantitatively reproducible, a higher dose of 'Acarbose' (500 mg) increasing the excretion ratio to about 1.0 indicating complete inhibition of intestinal sucrase activity. The suitability of the method for measuring differences in dose/response and duration of action was assessed by comparing three different alpha-glucosidase inhibitors (BAY g 5421, BAY m 1099, and BAY o 1248) and found to be satisfactory. Subjects with primary adult hypolactasia had urine lactose/lactulose excretion ratios raised to values indicating reduced rather than complete absence of lactase activity whereas sucrose/lactulose ratios were not significantly affected. 'Whole' intestinal disaccharidase activity assessed by this method demonstrated impairment of lactase, sucrase, and isomaltase in eight, one, and seven, respectively, of 20 patients with coeliac disease. By contrast in vitro assay of jejunal biopsy tissue indicated pan-disaccharidase deficiency in all but five of these patients. This shows the importance of distinguishing between 'local' and 'whole' intestinal performance. CONCLUSIONS: Differential urinary excretion of ingested disaccharides provides a reliable, quantitative, and non-invasive technique for assessing profiles of intestinal disaccharidase activity.

Acarbose↗

Combined assessment of intestinal disaccharidases in congenital asucrasia by differential urinary disaccharide excretion.

Investigation of intestinal disaccharide hydrolysis and permeability by means of a non-invasive differential sugar absorption test was performed in a family containing two siblings with primary sucrase-isomaltase deficiency. The procedure, which depends on measurement of urinary excretion ratios after the oral administration of lactose, sucrose, palatinose, lactulose and L-rhamnose, is capable of simultaneous determination of intestinal lactase, sucrase, and isomaltase activity and lactulose:rhamnose permeability. The results corresponded well with those of disaccharidase assay and histological findings in jejunal biopsy tissue obtained from the patients. Palatinose proved a satisfactory substrate for in vivo assessment of intestinal isomaltase activity. The method described provides a reliable and comprehensive assessment of intestinal disaccharide hydrolysis, and simultaneous estimation of permeability assists discrimination of primary from secondary disaccharidase deficiency. The ability to assess three different disaccharidase activities in addition to intestinal permeability by means of a single test, and the simplicity of preservation and transport of urine samples for sugar analysis, makes this a convenient, definitive method for the investigation of defective sugar absorption in both patients and population groups.

Child↗