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Mucosal Side (mucosal + side)
Selected AbstractsIncreased bacterial permeation in long-lasting ileoanal pouchesINFLAMMATORY BOWEL DISEASES, Issue 8 2006Anton J. Kroesen MD Abstract Background and Aims: Bacterial overgrowth appears to play an important role in the pathogenesis of ileoanal pouches. Therefore, the capability of bacterial permeation and its determinants is of great interest. The aim of this study was to examine bacterial permeation in the ileoanal pouch and to correlate the results with the degree of inflammation, the epithelial resistance, the mucosal transport function, and the age of the ileoanal pouches. Materials and Methods: Biopsies were taken from 54 patients before colectomy (n = 13; preileal pouch-anal anastomosis [IPAA]), and closure of ileostomy (n = 7; deviation), <1 year after closure of ileostomy (n = 8; intact pouch I), >1 year after closure of ileostomy (n = 16; intact pouch II), in the case of pouchitis (n = 11), and in 11 controls. Tissues were mounted in a miniaturized Ussing chamber. Escherichia coli was added to the mucosal side of the Ussing chamber, and the permeation was proven by serosal presence of E. coli. Epithelial and subepithelial resistance was determined by transmural impedance analysis. Active Na+ -glucose cotransport and active Cl, secretion were measured. Specimens were analyzed by fluorescent in situ hybridization with oligonucleotide probes targeting the bacterial 16s ribosomal RNA. The bacteria in and on the tissue were enumerated. Results: Bacterial permeation occurred in 2 of 13 pre-IPAA, 2 of 7 deviations, 0 of 8 intact pouch I, 9 of 16 intact pouch II, 5 of 11 pouchitis specimens, and 0 of 11 ileum controls. The frequency of bacterial permeation in the intact pouch II group is higher than in the intact pouch I group (P < 0.001). Epithelial resistance, mannitol fluxes, electrogenic chloride secretion, sodium-glucose cotransport of the bacterially permeated specimens versus nonpermeated of the intact pouch II group, and the pouchitis group and subepithelial resistance remained unchanged. Intramural bacteria could be detected by fluorescence in situ hybridization mainly in long-lasting pouches, but there was no correlation with bacterial permeation. Conclusions: The long-lasting ileoanal pouch is associated with increased bacterial permeability. This is not correlated with a disturbed function of the pouch mucosa but could be a precursor of pouchitis. [source] Influence of deoxynivalenol on the D -glucose transport across the isolated epithelium of different intestinal segments of laying hensJOURNAL OF ANIMAL PHYSIOLOGY AND NUTRITION, Issue 5-6 2007W. A. Awad Summary Deoxynivalenol (DON) decreases glucose absorption in the proximal jejunum of laying hens in vitro and this effect is apparently mediated by the inhibition of the sodium d -glucose co-transporter. DON could modulate the sugar transport of other intestinal regions of chickens. For this purpose, we have measured the effects of DON on the Na+d -glucose co-transporter, by addition of DON after and before a glucose addition in the isolated epithelium from chicken duodenum, jejunum, ileum, caecum and colon by using the Ussing chamber technique in the voltage clamp technique. The data showed in all segments of the gut that the addition of d -glucose on the mucosal side produced an increase in the current (Isc) compared with the basal values, the Isc after glucose addition to the small intestine was greater than the Isc of the large intestine compared with the basal values, specially of the jejunum (p < 0.002), indicating that the jejunum is the segment that is the best prepared for Na+ - d -glucose co-transport. Further addition of 10 ,g DON/ml to the mucosal solution decreased the Isc in all segments and the Isc returned to the basal value, especially in the duodenum and mid jejunum (p < 0.05). In contrast, the addition of 5 mmol d -glucose/l on the mucosal side after incubation of the tissues with DON in all segments had no effect on the Isc (p > 0.05), suggesting that DON previously inhibited the Na+d -glucose co-transport. The blocking effects of DON in duodenum and jejunum were greater than the other regions of the gut. It can be concluded that the small intestine of laying hens has the most relevant role in the carrier mediated glucose transport and the large intestine, having non-significant capacity to transport sugars, appears to offer a minor contribution to glucose transport because the surface area is small. The effect of d -glucose on the Isc was reversed by DON in all segments, especially in the duodenum and jejunum, suggesting that DON entirely inhibited Na+ - d -glucose co-transport. This finding indicates that the inhibition of Na+ co-transport system in all segments could be an important mode of action for DON toxicity of hens. Zusammenfassung Deoxynivalenol (DON) erniedrigt in vitro die Glukoseabsorption im proximalen Jejunum von Legehennen. Dieser Effekt ist vermutlich durch eine Hemmung des Natrium- d -Glukose-Cotransportsystems bedingt. DON könnte außerdem den Glukosetransport in anderen Segmenten des Darms beeinflußen. Zu diesem Zweck haben wir Wirkungen von DON auf das Natrium- d -Glukose-Cotransportsystem gemessen, indem wir DON nach und vor einer Glukosezugabe auf isolierte Darmepithelien des Duodenums, Jejunums, Ileums, Caecums und des Kolons mittels der Ussing-Kammer-Technik in der Volt-Klemmtechnik einwirken ließen. Die erzielten Daten wiesen in allen Segmenten des Darms verglichen mit den Basalwerten einen Anstieg im Strom (Isc) auf, wobei die Isc des Dünndarms bei Glukosegabe signifikant größer als die des Dickdarms waren, was darauf hinweist, dass das Jejunum am besten für den Glukosetransport geeignet war. Eine DON-Zugabe von 10 ,g/ml zur mukosalen Lösung schwächte den Isc in allen Segmenten, wobei die Isc speziell im Duodenum und mittleren Jejunum zum Ausgangswert zurück kehrten. Im Gegensatz dazu brachte die mukosale Glukosezugabe nach der DON-Inkubation keinen signifikanten Anstieg der Isc (p > 0,05), was auf eine durch DON hervorgerufene Blockade des Natrium- d -Glukose-Cotransportsystems schließen ließ. Es kann daraus geschlossen werden, dass der Dünndarm von Legehennen den bedeutendsten Einfluß im Glukosetransportmechanismus nimmt und der Dickdarm aufgrund einer kleineren Oberfläche einen geringeren Beitrag zum Glukosetransport leistet. Dem Isc steigernden Effekt der Glukose konnte signifikant durch DON in den Darmsegmenten besonders im Duodenum und im Jejunum entgegen gewirkt werden, was auf eine umfassende Hemmung des Natrium- d -Glukose-Cotransportsystems hinweist. Die Resultate weisen darauf hin, dass eine Hemmung des Natrium- d -Glukose-Cotransportsystems in allen Darmsegmenten eine wichtige Rolle in der DON-Toxizität für die Henne darstellen könnte. [source] Modulation of Na+ transport across isolated rumen epithelium by short-chain fatty acids in hay- and concentrate-fed sheepJOURNAL OF ANIMAL PHYSIOLOGY AND NUTRITION, Issue 11-12 2003S. K. Uppal Summary The effect of increasing concentrations of short-chain fatty acids [SCFA; mixture of the Na+ salts of acetic acid (62.5%), propionic acid (25.0%) and of butyric acid (12.5%)] on Na+ transport of sheep rumen epithelium was studied in vitro. The conventional Ussing chamber method was used for measuring Na+ transport rates (22Na+), short-circuit current (Isc) and tissue conductance (GT) of isolated rumen epithelium. SCFA in the buffer solution on the mucosal side caused a linear increase of Jnet Na+ from 1.14, to 1.22, 1.78 and 2.50 ,eq/cm2/h in hay-fed sheep at 0, 15, 40 and 80 mmol/l SCFA, respectively. In a second study, the effect of higher SCFA concentrations [0 (control), 80, 100 and 120 mmol/l] was investigated with epithelia from two groups of sheep. One group was subjected to hay ad libitum, whereas the other received concentrate feed (800 g/day in equal portions at 7.00 am and 3.00 pm) and hay ad libitum. Epithelia from concentrate-fed sheep again showed a significant (p < 0.05) and linear increase in Jnet Na+ at 80, 100 and 120 mmol/l. However, in hay-fed sheep, the difference in increase among 80, 100 and 120 mmol/l SCFA was not significant, indicating that, above 80 mmol/l SCFA Jms and Jnet exhibit saturation. Moreover, Na+ fluxes (Jms and Jnet) were generally higher in concentrate-fed than in hay-fed sheep at all SCFA concentrations and significant differences were observed at 100 and 120 mmol/l SCFA. The obtained results confirm the effect of SCFA on Na+ transport and are in agreement with studies regarding feeding regimes and electrolyte transport in the rumen. The important new observation is the increase of Na+ transport in concentrate-fed sheep even at high concentrations of SCFA (100 and 120 mmol/l). The enhanced activity of the Na+/H+ exchanger at these SCFA concentrations supports the assumption that the capacity for regulating the intracellular pH by extrusion of protons is increased, suggesting an adaptation in concentrate-fed sheep. This adaptation could prevent possible disturbances of epithelial functions (transport and barrier) under conditions of increased SCFA absorption. [source] The effect of short chain fatty acids on calcium flux rates across isolated rumen epithelium of hay-fed and concentrate-fed sheepJOURNAL OF ANIMAL PHYSIOLOGY AND NUTRITION, Issue 1-2 2003S. K. Uppal Summary The present in vitro experiment was conducted to study the effect of two concentrations of short chain fatty acids [SCFA: 0 (control), 40 and 100 mmol/l in the buffer solution on mucosal side] on calcium ion (Ca2+) transport across the isolated rumen epithelium of two groups of sheep. One group was subjected to hay ad libitum, whereas the other to concentrate feed (800 g per day in equal portion at 7.00 and 15.00 hours and hay ad lib). The conventional Ussing chamber method was used for measuring the Ca2+ transport rates (45Ca), short-circuit current (Isc) and tissue conductance (GT) of isolated rumen epithelium. The SCFA significantly increased Isc of the epithelia of concentrate-fed sheep. In both hay- and concentrate-fed animals, 45Ca flux rates showed an almost linear increase in net flux rate () with rising concentrations of SCFA, as a result of a combined effect of a large increase in mucosal-to-serosal flux rates () and an almost linear, but small, decrease in serosal-to-mucosal flux rate (). In concentrate-fed sheep and were significantly higher in tissues incubated with SCFA compared with hay-fed animals. The well-known adaptable morphological and functional changes in the rumen epithelium attributable to concentrate feeding obviously include Ca2+ transport; such feeding therefore may be considered as a possible prophylactic measure in the prevention of milk fever. [source] In vitro studies on the effects of Saccharomyces boulardii and Bacillus cereus var. toyoi on nutrient transport in pig jejunumJOURNAL OF ANIMAL PHYSIOLOGY AND NUTRITION, Issue 1-2 2000G. Breves The probiotics Saccharomyces boulardii and Bacillus cereus var. toyoi are nonpathogenic microbes which have been shown to affect certain functions of the mucosal barrier in pig jejunum such as electrogenic ion transport capacity and paracellular permeability. The present studies were performed to investigate potential effects of the probiotics on jejunal nutrient transport such as sodium-dependent glucose transport or proton-dependent dipeptide transport. For this purpose the in vitro Ussing-chamber technique was applied in order to examine net electrogenic ion flux rates (short circuit currents, Isc) across isolated intact jejunal epithelia in the absence and presence of either 10 mmol/l glucose (mucosal side) or two-fold application of 5 mmol/l glycyl- l -sarcosine or glycyl- l -glutamine to the mucosal bathing solution. Brush border membrane vesicles (BBMV) were prepared in order to characterize kinetic parameters (Vmax, Km) of Na-dependent glucose transport. Intestinal tissues were obtained from growing pigs in a weight range between 23 and 33 kg. All animals were fed twice daily and received 0.8,0.9 kg/day of a standard diet. After a 9- to 10-day adaptation period the diets for treated animals were either supplemented for 8 days with 1.7×107 colony-forming units (CFU)/g feed of S. boulardii or for 3 weeks with 106 CFU/g feed B. cereus var. toyoi. Under basal conditions Isc values were not affected by different treatment protocols (controls: 0.74 ± 0.04 µeq/cm2 per h, n=9; S. boulardii: 0.74 ± 0.12 µeq/cm2 per h, n=7; B. cereus 0.68 ± 0.09 µeq/cm2 per h, n=5). Irrespective of dietary treatment, the addition of glucose resulted in significant increases of Isc indicating substantial onset of electrogenic net Na/glucose cotransport. Maximal Isc values occurred within 30 min and reached 2.79 ± 0.41 µeq/cm2 per h in control epithelia. This was significantly lower than found in S. boulardii (4.47 ± 0.43 µeq/cm2 per h, p < 0.05) and B. cereus var. toyoi tissues (4.45 ± 0.31 µeq/cm2 per h, p < 0.05). Gt values were 22.4 ± 1.3 mS/cm2 in control animals and were significantly lower as shown in S. boulardii (p < 0.01) and B. cereus var. toyoi (p < 0.01)-treated animals (28.4 ± 1.3 and 29.9 ± 0.8 mS/cm2, respectively). Vmax values of Na-dependent glucose uptake into BBMV differed significantly between controls (0.64 ± 0.08 nmol/mg protein per 10 s; n=5), S. boulardii (0.89 ± 0.06 nmol/mg protein per 10 s; n=5, p < 0.05) and B. cereus var. toyoi preparations (1.08 ± 0.05 nmol/mg protein per 10 s; n=3, p < 0.01). Km values were not significantly affected (control: 0.31 ± 0.04 mmol/l, S. boulardii: 0.29 ± 0.05 mmol/l, B. cereus var. toyoi: 0.21 ± 0.01 mmol/l). Irrespective of dietary treatment, application of the dipeptide model substances glycyl- l -sarcosine or glycyl- l -glutamine resulted in significant increases of Isc indicating marked stimulation of electrogenic net H+/dipeptide cotransport. Highest Isc responses occurred in B. cereus var. toyoi preparations and lowest were found in control tissues. However, these differences were not significant. Gt values were not affected by different dietary treatments. The results clearly demonstrate that oral administration of either S. boulardii or B. cereus var. toyoi stimulates Na-dependent glucose absorption in pig jejunum. [source] Development of a New Tissue-Engineered Sheet for Reconstruction of the StomachARTIFICIAL ORGANS, Issue 10 2009Masato Araki Abstract We have developed tissue-engineered digestive tracts composed of collagen scaffold and an inner silicon sheet and successfully used it to repair defects in parts of the esophagus, stomach, and small intestine. However, some improvements were demanded for clinical usage because the silicon sheet presented technical difficulties for suturing and endoscopic removal. New tissue-engineered sheet (New-sheet) was composed of a single-piece and reinforced collagen scaffold with biodegradable copolymer. One beagle dog was used to evaluate whether New-sheet could withstand suturing in comparison with native digestive tracts using a tensile tester. Seven beagle dogs had a 5-cm circular defect created in the stomach. New-sheet soaked with autologous peripheral blood or bone marrow aspirate was sutured to the gastric wall. Endoscopic, histological, and immunohistochemical assessment was performed to evaluate regeneration of the stomach up to 16 weeks. Tensile strength testing showed that the mucosal side of New-sheet had strength almost equivalent to the mucosa of the esophagus (P = 0.61). Endoscopically, regeneration of the mucosa started from the circumference after 4 weeks, but a small linear ulcer was still evident at 16 weeks. The regenerated stomach shrank by 60,80% of its original size and histologically showed villous mucosa and underlying dense connective tissue. Immunohistochemically, the regenerated area expressed ,-smooth-muscle actin but was negative for basic calponin, irrespective of the source of soaked blood. New-sheet shows sufficient strength for suturing, no dehiscence, and better biocompatibility for clinical use, although further examination will be necessary to create a functional digestive tract. [source] | ||||||||||