Studies on L-arginase of the small intestine. I. Topographical distribution and some properties of the small intestine L-arginase in the rat.
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Small intestinal absorption of vitamin K2 was investigated in vitro. Experiments with increasing concentrations of the vitamin up to 900 nM revealed linerity between the concentration and the rate of absorption (r = 0.99). Addition of metabolic uncouplers and inhibitors such as 2,4-dinitrophenol, sodium azide, and potassium cyanide did not decrease the rate of absorption of the vitamin (P less than 0.05). Absorption rate of the vitamin increased when taurocholate was replaced by a nonionic detergent, Pluronic F-68. The addition of butyric and octanoic acids to the incubation solution caused an increase in the absorption rate of vitamin K2. No change in the absorption of the vitamin occurred in the presence of oleic and linoleic acid. Addition to vitamins K1 and K3 to the incubation solution did not change the rate of vitamin K2 absorption. These findings suggest that vitamin K2 is absorbed by the small bowel by a passive noncarrier-mediated diffusion process. The rate of diffusion varied when the lipid and bile salt composition of the incubation solution was modified. Distal intestinal absorption of vitamin K from bacterial sources coupled with colonic absorption of the vitamin may be the major constant source of vitamin K in mammals.
A method for measuring brush border membrane enzymes from small intestinal biopsies by crossed immunoelectrophoresis is presented. The use of a brush border specific antiserum made isolation of the brush border membrane before analysis unnecessary. This prevented loss of material which, together with inactivation of enzymes, was a limiting factor in previous studies of brush border enzymes from peroral biopsies. In 58 biopsies from patients without gastrointestinal disorders a close correlation between antigenic activity and corresponding enzymatic activity was shown for the following enzymes: sucrase-isomaltase (EC 3.2.1.48-EC 3.2.1.10), lactase-phlorizin hydrolase (EC 3.2.1.23-EC 3.2.1.62), microvillus aminopeptidase (microsomal, EC 3.4.11.2) and dipeptidyl peptidase IV (EC 3.4.14.X). The immunoelectrophoretic patterns of intestinal mucosa near the ligament of Treitz, and in jejunum and ileum were established. The method presented is thought to be of value in further studies of the molecular basis of brush border diseases.
The activities of the digestive enzymes, maltase [EC 3.2.1.20], sucrase [EC 3.2.1.26], trehalase [EC 3.2.1.28], Leucine aminopeptidase [EC 3.4.11.1], and alkaline phosphatase [EC 3.1.3.1] were measured in various regions of the small intestine of rats. The activities of all these enzymes were much higher in the jejunum than in the ileum, and in the distal regions of the ileum no sucrase, trehalase or alkaline phosphatase activity was detected. In the jejunum, the activities of all the enzymes tested exhibited clear circadian variations with the highest activity at 0000-0400 h and the lowest at 1200 h when the rats were fed ad libitum. In the ileum, maltase and sucrase also exhibited circadian variations, but the amplitude of the rhythm was smaller than that in the jejenum. Trehalase and alkaline phosphatase did not show any circadian variation in the ileum. Leucine aminopeptidase showed a circadian variation in the ileum with the same amplitude as in the jejunum. The phase of the circadian variations shifted about half a day when the rats were fed in the daytime, but the amplitude of the rhythm did not change.
The enlargement of the small intestine of mice during lactation is due, at least in part, to hyperplasia in the mucosal crypts and villi. The number of cells per crypt increases by 130% and the cell production rate by 63% after 15 days of lactation. These parameters were measured from crypt squashes and sections using both double-label and PLM techniques. Neither the numbers of crypts and villi in the small intestine nor the turnover time of post-mitotic cells on the villi changed. A number of factors appear to act in concert during lactation to trigger this increase in epithelial cell number in the small intestine. The experiments reported suggest a role for the increased quantity of food consumed by the lactating animal, for changing hormonal levels, and for the increased demands placed on the body by milk production.
An absorption screen was performed in 10 chronic alcoholic patients within a few days of admission due to an acute alcoholic episode. Impaired absorption of d-Xylose was noted in three patients and low leucocyte ascorbic acid and serum folate levels in five. No abnormality was detected in jejunal histology. The absorption of water and electrolytes from the jejunum was studied in these patients using a triple-lumen tube perfusion system. The mean rate of absorption of water in the alcoholic subjects (50-0 +/- 2-3 ml/h) was significantly lower (P less than 0-001) than the mean value in 14 healthy control subjects (205 +/- 15-9 ml/h). A significant reduction of Na+ and Cl-absorption was also demonstrated in the alcoholic subjects. These results indicate that patients with acute-on-chronic alcoholism may have a function impairment of water and electrolyte absorption from the jejunum. This may, in part, account for some of the nutritional deficiencies in such patients and for symptoms such as diarrhoea which may be present.
Small intestine neuroendocrine tumors (SI-NETs) frequently present as multifocal primaries. We commonly observe microscopic lesions in the superficial layer of the small intestine of SI-NET patients. We aimed to define them as small intestinal neuroendocrine tumorlets (SINTs) and explore their clinical and biological significance. Twenty multifocal and twenty unifocal SI-NETs patients who received resection at a single institution were retrospectively reviewed. Four hundred and forty six archived pathological slides were examined for microscopic lesions located in the lamina propria, muscularis mucosa, and superficial submucosa. Clinicopathological associations and progression-free survival were analyzed. Previously published genomic data were re-analyzed. SINTs were identified in 50% of multifocal and 30% of unifocal SI-NET patients. Median SINT size was 95 μm, with a median distance of 2.2 mm from the nearest mass. Compared to the 'true unifocal' group (unifocal without SINT), the 'multifocal-spectrum' group (multifocal or unifocal with SINT) had higher BMI (median: 27.6 vs 22.8, P = 0.0060), higher rates of perineural invasion (OR: 5.5, P = 0.044), larger mesenteric mass (median: 2.6 vs 1.6 cm, P = 0.034), and more advanced pT stage (pT3 or pT4, OR: 7.2, P = 0.018). Genomic re-analysis suggested that 13% of cells in multifocal primary tumors could share clonal origins, possibly indicating clonal spread via SINTs. SINTs may serve as a new biomarker for multifocal spectrum with local aggressiveness. The actual frequency of multifocal SI-NET may be higher than currently recognized in clinical practice. Further studies are needed to validate their prognostic and biological significance.
Total and specific lactase activities in the small intestine of Chester White and Hampshire pigs were measured at 1, 8, 15, 22, 29, and 43 days of age. The small intestine was divided into 10 segments of equal length, the proximal 10 cm of each segment was scraped, and the scrapings were homogenized for use in the lactase determinations. Significant breed, age, and segment differences were observed for both specific and total activities. In both breeds, the total lactase activity at 1 day of age was lower than that at any other age. After reaching maximal levels at 15 days of age, the total activity declined with the loss of activity occurring primarily in the ileum. At 1 and 8 days of age, the total lactase activities for the two breeds were similar, but the Chester White pigs had higher activities at all other ages. The pattern of changes in specific activity with age was similar for both breeds. The mean specific activity was highest at 1 and 8 days of age and then fell progressively to minimal levels at 43 days of age. Chester Whites had higher specific activities than Hampshires during the first 4 weeks of life, but at 6 weeks of age there was little difference between the breeds. The peak lactase activity, expressed as total or specific activity, occurred in the proximal one-third of the small intestine of both breeds, and the distal one-third of the gut had relatively low activities as the animals matured.
To study the effect of intraperitoneal S(-)sulpiride (1-15 mg/kg), R(+)sulpiride (5-10 mg/kg), metoclopramide (1-15 mg/kg), cisapride (10 mg/kg) and domperidone (5-10 mg/kg) on intestinal progression, rats were given the test drug followed by oral lactulose. Their hydrogen excretion was used to calculate the small bowel transit time (SBTT) and maximum peak time (MPT). Metoclopramide (7.5 mg/kg) had the greatest effect on SBTT (-25%), followed by S(-)sulpiride and domperidone. S(-)sulpiride (10 mg/kg) had the greatest activity on the MPT (-35.2%) followed by metoclopramide. R(+)sulpiride and cisapride did not modify SBTT and MPT. In conclusion S(-)sulpiride is the isomer active on intestinal transit and DA2-receptors seem important targets in the modulation of intestinal progression, since S(-)sulpiride, metoclopramide and domperidone are DA2-receptor antagonists, and R(+)sulpiride and cisapride are not. The H2 breath test proved a valid method for measuring the effect of drugs on the small intestine in animals.
The mechanismus of active transport in the small intestinal mucosa are discussed. A survey on the different defects of disturbed absorption is given and examples of 3 basic types of malabsorption are discussed.
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An influx of fluid into the lumen of the intestine similar to that seen in simple obstructional ileus may be provoked by introducing a hyperosmolal glucose solution into the bowel. In the otherwise intact small intestine the effect of this influx of fluid will be in accordance with a simple dilution curve. The intestinal mucosa thus functions in the manner of a semipermeable membrane permitting only hypo-osmolal fluids to enter the intestinal lumen and in amounts independent of parenteral fluid infusions, regardless of osmolality. This relationship persists even after the intestine has been totally obstructed for 3 days. The influx of fluid has the same principal characteristics, and the only limiting factor on the magnitude of this fluid shift to the intestine is the lack of fluids resulting from the marked dehydration of the organism due to ileus. Prerequisites for this are normal epithelial function and normal mucosal circulation. Thus it is clear that the organism in general and the small intestine in particular, even when exposed to prolonged obstruction, are still able to counteract intraluminal hyperosmolality by dilution with hypo-osmolal fluid.
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We have used the perforated-patch whole-cell recording mode of the patch-clamp technique to monitor membrane potential and measured cell volume changes by image analysis, to determine the nature of the response to secretagogues of isolated whole guinea-pig small-intestinal crypts. Vasoactive intestinal polypeptide (VIP) produced a dose-dependent depolarisation (EC50 = 30 nM) and an increase in membrane conductance that could be potentiated by carbachol. Similar depolarisations were observed with forskolin. The depolarisation induced by 100 nM VIP was smaller when pipette [Cl-] was 60 mM than when it was 145 mM, suggesting an effect through Cl- conductance activation. Carbachol alone produced a hyperpolarisation (EC50 = 2 microM). The Cl- channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB) produced a small hyperpolarization. When VIP was added in the presence of NPPB, the depolarisation was observed instead, consistent with the parallel activation of a K+ conductance. Both carbachol (100 microM) and VIP (100 nM) induced a 25%-30% shrinkage of crypts, which was maximal 8 min after addition of the secretagogue. The induced shrinkage was sustained in the continued presence of agonist and was reversed upon washout. Shrinkage induced by the agonists was abolished by increasing extracellular K+ from 6 mM to 20 mM and was inhibited partially in the presence of 100 microM anthracene-9-carboxylic acid in the bath. The decrease in volume induced by 100 nM VIP was totally abolished in the presence of 100 microM NPPB. The results are consistent with the view that both VIP and carbachol induce secretion in small-intestinal crypts.
Previous data have shown that the life span of small intestine epithelial cells in germ-free (GF) mice was 4.3 days, while that in conventional (CV) mice was 2.1 days, under ad libitum feeding. On the other hand, in the author's laboratory, it was also found that feeding conditions affected the cells' life span. That is, in CV mice the life span of the cells lengthened under restricted feeding (2.6 days), compared with under and libitum feeding (1.8 days). In the present experiment the life span of small intestine epithelial cells was investigated using radioautography, under controlled feeding (setting it equal to ad libitum feeding) and restricted feeding, in both CV and GF mice. Small intestine samples were taken from the middle part of duodenum, jejunum and ileum. Body weight changes, organ wet weights and intestine were also measured. In the lower part of the small intestine the effects of a restricted diet on epithelial cell life span prolongation appeared clearly in CV mice, but this effect was reduced in GF mice. This may be partly because the restricted group had slightly shorter villi in the case of GF mice.