Metabolic development of the small intestinal mucosa in rodents.
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Animals↗
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[Proceedings: Structural and functional aspects of pathologic pictures of the small intestine mucosa].
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Biopsy↗
The early transcriptional response of pig small intestinal mucosa to invasion by Salmonella enterica serovar typhimurium DT104.
Salmonella enterica serovar typhimurium (S. typhimurium) species are a leading cause of human invasive gastroenteritis. There is increasing in vitro evidence about Salmonella interaction with isolated cells or cell lines (macrophages, and enterocytes) on the molecular level, however, very little is known about in vivo interactions during actual invasion. We investigated the early interaction of S. typhimurium with intact small intestinal mucosa, in a pig model. Intestinal segments were infected with or without S. typhimurium DT104, and perfused. Whole mucosal gene expression was analyzed by cDNA array on 0, 2, 4, and 8h post-infection. Invasion resulted in the upregulation of only eight transcripts in jejunal mucosa, among those the proinflammatory IL-8 (at 4h only), and the antiinflammatory STAT3 (at 4 and 8h). The limited number of differentially expressed genes found here in vivo compared to in vitro is most likely due to the presence of multiple, heterogenous cell interactions in intact mucosa. Furthermore, it is concluded that S. typhimurium evades strong host responses by downregulating the local inflammatory response.
Animals↗
Eicosanoids content in small intestinal mucosa of children with celiac disease.
Celiac disease (CD) is characterized by diarrhea, growth retardation, and weight loss in genetically susceptible subjects on a gluten-containing diet. The exact pathogenesis of CD is still obscure, but it is considered to be immunologically mediated. We have previously shown elevated prostaglandin E2 (PGE2) and thromboxane B2 (TxB2) content in small intestinal mucosa obtained from active celiac children. In the present study, we found significantly elevated PGE2, leukotriene B4 (LTB4), and leukotrienes C4, D4, and E4 (LTC4D4E4) content in small bowel mucosa from children suffering from CD on a gluten-containing diet in comparison to control subjects. PGE2 was 25,278 +/- 7,761 vs. 4,478 +/- 426 pg/mg of protein (mean +/- SEM), respectively. LTB4 was 8,807 +/- 3,706 vs. 403 +/- 63 pg/mg of protein (mean +/- SEM), respectively. LTC4D4E4 was 15,369 +/- 4,085 vs. 2,998 +/- 279 pg/mg of protein (mean +/- SEM), respectively. We conclude that the elevated content of arachidonic acid metabolic products via cyclooxygenase and lipoxygenase pathways may contribute to the diarrhea and may be involved in the pathogenesis of mucosal injury.
Adolescent↗
[Histological studies on cow intestinal mucosa and electron microscopic studies of the ruminal epithelium of cows fed a deficient diet for a prolonged time].
Histological and histochemical studies were carried out on the gastro-intestinal mucosa of three experimental cows (roughages/maize silage) and three control animals. The animals were slaughtered on termination of the long-term trial. Mucosa samples were taken, for further study, from the rumen, the duodenum, the jejunum, the large intestine and the appendix. Histochemical analysis did not reveal any essential differences in the activities of non-specific esterase, alkaline and acid phosphatase and lactic acid dehydrogenase in the mucosa of the rumen, the large and the small intestine and the appendix of both the experimental animals and the controls. The experimental animals were found to exhibit a higher rate of glutamate-dehydrogenase activity in the ruminal mucosa and in the mucosa of the large and small intestine. A higher succinate dehydrogenase activity was observed in the ruminal mucosa of the experimental animals, relative to that of the controls, while the activity in the intestinal mucosa was decreased. Only slight changes were noted in the activity of the enzymatic systems tested. Electron microscopic studies did not reveal any differences in the ultrastructure of the epithelial cells of the ruminal mucosa in both the experimental animal and the controls.
Acid Phosphatase↗
The palmitic acid binding properties of cytosolic proteins located in the villus and crypt zones of bovine intestinal mucosa.
The palmitic acid binding capacity of cytosolic proteins in three preparations obtained by differential scraping of bovine intestinal mucosa were compared. The data indicated that the palmitic acid binding activities depended on the position that the cells occupied along the crypt-villus axis, as shown from the level of alkaline phosphatase activity. Proteins with palmitate binding properties in the high- and low-molecular-weight cytosolic proteins in the villus zone bound 1.24 +/- 0.41 and 1.54 +/- 0.16 pmol palminate/micrograms protein respectively. The binding decreased to 0.50 +/- 0.25 and 1.10 +/- 0.23 pmol palmitate/micrograms for the proteins in the crypt zone. Ammonium sulphate fractionation and gel filtration chromatography indicated that the low-molecular-weight cytosolic proteins obtained from light mucosal scrapings contained the highest palmitate binding activity. These results suggest that the cytosolic proteins located in the villus zone may play a role in the absorption of fatty acids.
Alkaline Phosphatase↗
Biosynthesis of glycoproteins in the intestinal mucosa. II. Influence of diets.
To determine the effect of diets on the biosynthesis of glycoproteins in the intestinal mucosa, male Sprague-Dawley rats were fed, ad libitum, high-carbohydrate high-fat and high-protein diets. In in vivo methodology, the animals from each dietary group received an intraperitoneal injection of [14C]-fucose, or [14C]-galactose or [14C]-glucosamine. Incorporated radioactivity was detected in the different subcellular fractions. In in vitro methodology, five glycosyl-transferases were studied on microsomes and cell sap. In vivo, only the high-fat diet induces a decrease in the incorporation of [14C]-glucosamine in the macromolecules. In vitro, only the fucosyl-transferase activity is modified by several diets: as compared with an increase in the activity of the control with the age of the animals, high-protein diet induces a slight activation after 2 weeks of diet, while high-carbohydrate and particularly high-fat diets inhibit the enzyme activity. The inhibition is the same when high-fat diet consists of either oil mixture or triolein or oleic acid or linoleic acid and becomes significant with a supplementation rate of 10%.
Animals↗
Factors stabilizing pyrroline-5-carboxylate synthase of rat intestine mucosa at a physiological temperature.
Mammalian pyrroline-5-carboxylate (PC) synthase in the mitochondrial membrane of rat small intestine mucosa possesses marked thermal instability at temperatures of 30 to 37 degrees C [Y. Wakabayashi, J. G. Henslee, and M. E. Jones (1983) J. Biol. Chem. 258, 3873-3882]. Factors stabilizing the enzyme activity at 37 degrees C were extensively examined by incubating the enzyme with various compounds before assay. In the presence of 60% sorbitol, the enzyme retained full activity for 30 min. Xylitol, glycerol, and fructose were also effective, although sucrose, ethylene glycol, polyethylene glycol and dimethyl sulfoxide were ineffective. AMP, GMP, IMP, and UMP (15 mM) were completely protective while ATP and adenosine were not. Phosphate and arsenate at 10 mM maintained 90 and 82%, respectively, of the original activity after 10 min. NADPH and NADP (3 mM) were protective whereas 3 mM NADH was not. The possibility that phosphate and NADPH are stabilizing PC synthase in vivo was discussed. Addition of 0.13 mM p-chloromercuriphenylsulfonic acid or 0.55 mM 5,5'-dithiobis-(2-nitrobenzoic acid) to the enzyme resulted in complete loss of activity, but prior addition of excess dithiothreitol to the enzyme prevented the inactivation, suggesting that a sulfhydryl group is involved in the activity.
Adenosine Monophosphate↗
Whipple's disease: a histological, immunocytochemical and electronmicroscopic study of the immune response in the small intestinal mucosa.
Whipple's disease is a multisystem disorder with protean manifestations and with poorly understood aetiopathogenesis. It is unclear how the immune system reacts, whether it functions normally or not, whether it protects the organism or is defeated in one way or another by the 'Whipple bacillus'. The purpose of our study was to assess humoral and cellular immunity at the level of the intestinal mucosa. This histochemical, immunocytochemical and electronmicroscopic study, based on 16 cases, has shown that the changes in components of the mucosal immune system in Whipple's disease are quite different from normal. The phagocytic capacity of the macrophages, assessed microscopically, is abnormal, the number of intra-epithelial lymphocytes is increased, the CD 4/CD 8 cell ratio is decreased and the IgM positive cells in the lamina propria outnumber the IgA positive cells. These changes may be inter-dependent.
Adult↗
[Immunohistologic changes of the intestinal mucosa in chronic inflammatory bowel diseases].
In the pathogenesis of inflammatory bowel disease abnormally increased immune reactions in the intestinal mucosa play a basic role. The revealing of these reactions was facilitated by the rapidly spreading immunohistological methods in last decades. This review discusses in detail the characteristic distribution of lymphocyte subsets and plasma cells in Crohn's disease and ulcerative colitis. Results indicating increased expression of adhesion molecules are also presented, as well as the different patterns of cytokine production in Crohn's disease and ulcerative colitis. The rapid epithelial turnover developing as a consequence of increased rate of proliferation and apoptosis in inflammatory bowel disease is also shown. The epithelial expression of MHC II antigen in colonic mucosa which is not observed in healthy subjects is also discussed. In addition, the pathogenetic role of lost oral tolerance in the development of inflammatory bowel is reviewed in detail.
Chronic Disease↗
Lymphocyte phenotypes in the intestinal mucosa of sheep infected with Trichostrongylus colubriformis.
Monoclonal antibodies to ovine lymphocyte surface antigens were used in an immunohistochemical study of the intestine of sheep. In the epithelium CD8+ cells predominated whereas the majority of lamina propria T lymphocytes were CD4+. Infection of sheep with the parasitic nematode Trichostrongylus colubriformis including sufficiently large numbers of parasites to induce protective immunity did not alter the number of CD4+ or CD8+ lymphocytes in the intestinal mucosa. In contrast, exposure of naive sheep to a single large infection of T. colubriformis resulted in a substantial decrease in number of CD8+ cells and moderate decreases in number of CD4+ cells in the duodenal but not the jejunal mucosa. MHC class II antigens were not detected either in or on epithelial cells of the sheep small intestine.
Aging↗
Interaction of a self-emulsifying lipid drug delivery system with the everted rat intestinal mucosa as a function of droplet size and surface charge.
PURPOSE: To investigate the interaction of positively charged self-emulsifying oil formulations (SEOF) following aqueous dilution as a function of resulting emulsion droplet charge and size with rat everted intestinal mucosa, adherent mucus layer and Peyer's patches, using cyclosporine A (CsA) as a lipophilic model drug. METHODS: Droplet size determination (TEM technique) and zeta-potential measurements were used to characterize the resulting emulsions. For the ex vivo interaction study, the well-known rat intestine everted sac technique was used in combination with confocal microscopy. RESULTS: The positively charged oil droplets formed by SEOF dilutions at ratios of 1/50 and 1/10 elicited the stronger interaction with the mucosal surface. The positive charge of the smaller droplets was more readily neutralized, and even reversed in aqueous solutions containing moderate subphysiological mucin concentrations. Parameters such as droplet size, negativity of the epithelial mucosa potential and presence of the mucus layer on the epithelial surface affected drug mucosa uptake and the adhesion of the positively charged droplets to the rat intestinal mucosa. CONCLUSIONS: The enhanced electrostatic interactions of positively charged droplets with the mucosal surface are mostly responsible for the preferential uptake of CsA from the positively charged droplets as compared to negatively charged droplets irrespective of the experimental conditions used. The increased uptake of the CsA from the negatively charged oil droplets was consistent with the dilution extent, as expected, whereas in the positively charged droplets, an intermediate droplet size range was identified resulting in optimum drug uptake and clearly suggesting that drug uptake was not consistent with either dilution extent or droplet size.
Animals↗
Bioadhesion of lectin-latex conjugates to rat intestinal mucosa.
PURPOSE: The specific interactions between three lectin-latex conjugates and different structures of rat intestinal mucosa have been studied ex vivo. METHODS: These systems were prepared by covalent coupling of different ligands, i.e., tomato lectin (TL), asparagus pea lectin (AL), mycoplasma gallisepticum lectin (ML), and bovine serum albumin (BSA) as control, to poly(styrene) latexes. RESULTS: Using mucosa samples without Peyer's patches (PP), the extent of interaction of all three lectin-latex conjugates with the mucosa decreased from duodenum to ileum, probably due to progressive diminution of the mucin concentration along the gastrointestinal tract. The following order of interaction of the conjugates with the mucus gel layer was observed: TL > ML = AL (p < 0.05). For each lectin, these results corresponded well to the concentration of its specific sugar in the mucus. Using intestinal samples with PP, an important increase of interaction of the conjugates with the mucosa was found for ML (about 25%) and AL (about 50%), whereas the interaction of TL decreased about 25%. CONCLUSIONS: Photomicrographs with fluorescent latexes have confirmed the specificity of the ML- and AL-latex conjugates for the PP region and of the TL-latex conjugates for the mucus gel.
Adhesives↗
An in situ single-pass perfusion model for assessing absorption across the intestinal mucosa of the brushtail possum.
AIM: To develop an in situ animal model for assessing absorption of molecules across the intestinal mucosa of possums. METHODS: A surgical preparation was used to perfuse known concentrations of reference compounds (fluorescein and luteinising hormone-releasing hormone; LHRH) through measured sections of selected regions (jejunum, caecum, proximal colon) of the intestinal tract of 19 possums, over a 2-h period. Plasma concentrations of the compounds, which were perfused either with or without co-administration of a permeation enhancer (sodium deoxycholic acid; SDA), were determined in the perfusion effluent, peripheral and in some instances in the pre-hepatic circulation by spectrofluorometry (fluorescein) or radioimmunoassay (LHRH). Pharmacokinetic parameters of both compounds in the possum were determined over a period of up to 4 h in a further 30 animals (fluorescein, n = 6; LHRH n = 24), from their plasma profiles following intravenous (I/V) administration of a bolus dose. RESULTS: In animals perfused with 25 mg/ml fluorescein (Perfusion Experiment (PE) 1), the mean plasma concentration was 2.8 (SE 0.12) microg/ml in post-hepatic blood samples. When possums were perfused with 2.5 mg/ml fluorescein and 7 microg/ml LHRH (PE 2), mean plasma concentrations were 0.3 (SE 0.01) and 7.8 (SE 1.64) microg/ml fluorescein and 0.1 (SE 0.02) and 6.3 (SE 0.45) ng/ml LHRH, in the absence and presence of permeation enhancer, respectively. There was a poor correlation between pre-hepatic and post-hepatic concentrations. CONCLUSIONS: The single-pass in situ perfusion technique provided a useful model for investigating basic information on the absorption of biocontrol agents across the intestinal tract of possums, but had limitations that must be recognised.
Animals↗
Influence of surface-active food additives on the integrity and permeability of rat intestinal mucosa.
The influence of two surface-active food additives on the integrity and permeability of rat ileal mucosa has been studied. We determined the activity of N-acetyl-beta-glucosaminidase, a lysosomal enzyme, in the rat intestinal lumen after deposition of polyoxyethylene (20) sorbitan monostearate (polysorbate 60; Tween 60) or polyoxyethylene (20) sorbitan monooleate (polysorbate 80; Tween 80) in a section of ligated, cannulated gut. We also determined the activities of N-acetyl-beta-glucosaminidase, alkaline phosphatase, 5'-nucleotidase and phospholipase A2 in mixtures of isolated mucosal cells and polysorbate 60 or polysorbate 80. The activity of N-acetyl-beta-glucosaminidase was increased in the luminal contents of the cannulated gut 15 min after deposition of either polysorbate 60 or polysorbate 80 (10 mg/ml fluid instilled into gut). It was also increased in mixtures of mucosal cells and polysorbate 60 or polysorbate 80 (0.1-10 mg/ml). In contrast, the activities of alkaline phosphatase and 5'-nucleotidase were unaffected and that of phospholipase A2 was decreased by the presence of either polysorbate. These findings indicated that polysorbate 60 and polysorbate 80 released lysosomal enzymes from the intestinal mucosal cells and that these agents might damage the intestinal mucosa and increase its permeability. We therefore determined the intestinal permeability to sodium fluorescein in the absence and presence of polysorbate 60 or 80 and found that the permeability was slightly increased in the presence of either of the compounds at concentrations of 10 mg/ml fluid instilled into gut. It is possible therefore that surface-active food additives might impair the function of the mucosal barrier and increase the permeability of the gut to potentially toxic and pathogenic molecules.
5'-Nucleotidase↗
T cell receptor repertoire in the peripheral blood and intestinal mucosa of coeliac patients.
The alpha beta and gamma delta T cell receptor (TCR) repertoire in the peripheral blood and intestinal mucosa of six coeliac and six age-matched controls was analysed by reverse transcription and polymerase chain reaction (PCR). No TCR alpha and gamma delta restriction was observed in coeliacs and controls. However, V gamma 3 was expressed only in coeliac peripheral and intestinal T cells. V delta 2 was strongly expressed in coeliacs and scarcely transcribed in control cells. The unique expression of these gamma delta TCR in coeliac patients suggests that V gamma 3 and perhaps V delta 2 TCR-bearing lymphocytes may play a role in the pathogenesis of coeliac disease.
Celiac Disease↗
[New pathogenetic aspects of hepatitis B in relation to finding HBAg inhibitor in the intestinal mucosa and human feces].
Hepatitis B antigen (HBAg) positive sera became negative after in vitro incubation with homogenates of human faeces or intestinal mucosa. This was found to occur in all sera tested by various methods. These findings suggest the existence in the human intestine of a substance able to inactivate the HBAg and that it is not an antibody or of the nature of interferon. The presence of an inhibitor could explain why B-type hepatitis is seldom if ever faecally transmitted and also the low oral infectiousness of the B virus.
Antibodies, Viral↗
[Enzymatic activity of the pancreas and small intestinal mucosa in modelling of toxic hepatitis by administration of heliotrine].
The activity of pancreatic (alpha-amylase, protease, and lipase) and enteral (maltase, glycyl-L-leucine dipeptidase, monoglyceride lipase) was studied in the experiments on adult male Wistar rats weighting 350 +/- 35 g 1-3, 7, 10, 30, 60, and 90 days after injection of a single dose of 25 mg per 100 g body weight. Acute heliotrine intoxication (in a toxic hepatitis model) was shown to lead to a marked decrease in the activities of both pancreatic and intestinal enzymes. It is suggested that the changes may be associated with the direct action of heliotrine on the pancreas and small intestinal mucosa or with hepatic failure.
Animals↗