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PubMed1999

Activities of enzymes involved in glutamine metabolism in connection with energy production in the gastrointestinal tract epithelium of newborn, suckling and weaned piglets.

Changes in the activity of enzymes involved in glutaminolysis and energy metabolism in the entire gastrointestinal (GI) tract of developing piglets are presented for the first time. The activities of glutaminase, glutamate dehydrogenase, oxoglutarate dehydrogenase, isocitrate dehydrogenase and alanine aminotransferase in the epithelium along the gastrointestinal tract from newborn, suckling (2-4 weeks old) and weaned (9 weeks old) piglets were investigated. The activity of glutaminase in the epithelium from the small intestine and colon was higher (p < 0.05) in weaned piglets than in newborn and suckling piglets. In addition, glutamate dehydrogenase and alanine aminotransferase activities in the small intestinal epithelium were higher (p < 0.05) for weaned piglets than for newborns. The activity of oxoglutarate dehydrogenase in the epithelium of the small intestine was significantly lower in newborn and suckling piglets compared with weaned individuals. The activity of isocitrate dehydrogenase in the epithelium along the gastrointestinal tract was higher (p < 0.05) for suckling and weaned piglets than for newborn piglets. The present data indicate that the utilization of substrates for energy production differs markedly between the stomach, small intestine and colon of growing piglets. Also, the capacity of enzymes in the epithelium of the GI tract to utilize acetyl-CoA as an energy substrate in the tricarboxylic acid cycle increased with piglet age. The epithelium of the GI tract of the newborn, suckling and weaned piglets showed a high capacity to metabolize alpha-ketoglutarate.

Aging↗
PubMed2006

Diagnosis and surgical management of vascular ectasia in a dog.

CASE DESCRIPTION: An 8-year-old male Golden Retriever was evaluated because of an 8-week history of intermittent diarrhea with melena and hematochezia that were not responsive to medical treatment and resulted in severe anemia. CLINICAL FINDINGS: Exploratory celiotomy with intestinal and colonic biopsy revealed mild enterocolitis but did not result in diagnosis of the cause of melena and hematochezia. Endoscopy of the upper portion of the gastrointestinal tract and colonoscopy were performed. Multifocal areas of coalescing, tortuous mucosal blood vessels were observed in the cecum and all regions of the colon. A diagnosis of vascular ectasia (VE) was made on the basis of the endoscopic and histologic appearance of the lesions. TREATMENT AND OUTCOME: An ileorectal anastamosis was performed. Melena and hematochezia resolved within 3 days after surgery, and the anemia resolved within 6 weeks after surgery. Surgical resection of the cecum and colon and feeding of a highly digestible diet resulted in long-term (22 months) resolution of clinical signs. CLINICAL RELEVANCE: Initial exploratory celiotomy with intestinal and colonic biopsy failed to reveal the VE lesions responsible for the melena, hematochezia, and anemia. Endoscopic evaluation was necessary for detection of the colonic VE lesions. Surgical resection of the cecum and colon and feeding of a highly digestible diet may result in a favorable outcome in affected dogs.

Anastomosis, Surgical↗
PubMed2001

Is intestinal ischemia a risk of laparoscopy? An experimental study in rabbits.

BACKGROUND AND PURPOSE: In the literature, there are few reports documenting intra-abdominal organ necrosis following laparoscopic procedures. This experimental study was planned to investigate whether intestinal ischemia develops during laparoscopic procedures and if laparoscopy could cause intestinal necrosis. MATERIAL AND METHODS: Two experimental groups, each consisting of 10 adult New-Zealand rabbits, were used in this study. The first group comprised the study group which had pneumoperitoneum, the second group comprised the normal animals serving as the controls. A cervical tracheostomy was performed to achieve successful general anesthesia in both groups. In the study group, intraperitoneal CO2 insufflation was carried out and intraabdominal pressure (IAP) was adjusted so as not to exceed the arterial blood pressure. After 20 min high IAP period, the intraabdominal gas was aspirated. Five minutes later, samples of both small intestine and colon tissue were taken. In the control group, tissue samples were taken 25 min after anesthesia was achieved. Xanthine oxidase (XO) and malondialdehyde (MDA) levels were measured as indicators of intestinal ischemia and lipid peroxidation in the intestinal tissues. Statistical analysis was done to compare the XO and MDA levels of the small intestines and colons of both groups. RESULTS: The mean colonic XO levels were 1.323+/-1.17 and 0.217+/-0.27 (U/mg protein) in study and control groups, respectively. This difference was statistically significant (t = 2.60, p<0.05). The other comparisons with regard to XO and MDA levels were statistically not significant. CONCLUSION: Our results demonstrate that intraperitoneal CO2 insufflation in which intraabdominal pressure was adjusted to be lower than arterial blood pressure may affect oxygenization of the colon.

Animals↗
PubMed1987

In vivo colonization of the mouse large intestine and in vitro penetration of intestinal mucus by an avirulent smooth strain of Salmonella typhimurium and its lipopolysaccharide-deficient mutant.

The relative abilities of an avirulent Salmonella typhimurium strain with wild-type lipopolysaccharide (LPS) character, SL5319, and a nearly isogenic LPS-deficient mutant, SL5325, to colonize the large intestines of streptomycin-treated CD-1 mice in vivo and to penetrate colonic mucus in vitro were studied. Previously it had been shown that, when fed simultaneously to streptomycin-treated mice (approximately 10(10) CFU each), the S. typhimurium strain with wild-type LPS colonized at 10(8) CFU/g of feces indefinitely, whereas the LPS-deficient mutant dropped within 3 days to a level of only 10(4) CFU/g of feces. In the present investigation, when SL5325 was allowed to colonize for 8 days before feeding mice SL5319 or when it was fed to mice simultaneously with an Escherichia coli strain of human fecal origin (10(10) CFU each), both strains colonized indefinitely at 10(7) CFU/g of feces. Moreover, when the wild-type and LPS-deficient mutant strains were fed to mice simultaneously in low numbers (approximately 10(5) CFU each) the strains survived equally well in the large intestines for 8 days, after which the LPS-deficient mutant was eliminated (less than 10(2) CFU/g of feces), whereas the wild-type colonized at a level of 10(7) CFU/g of feces. In addition although both strains were able to adhere to mucus and epithelial cell preparations in vitro, the wild-type strain was shown to have greater motility and chemotactic activity on CD-1 mouse colonic mucus in vitro and to more rapidly penetrate and form a stable association with immobilized colonic mucosal components in vitro. Based on these data, we suggest that the ability of an S. typhimurium strain to colonize the streptomycin-treated mouse large intestine may, in part, depend on its ability to penetrate deeply into the mucus layer on the intestinal wall and subsequently, through growth, colonize the mucosa.

Animals↗
PubMedDate not supplied

Cytolytic capabilities of lamina propria and intraepithelial lymphocytes in normal and chronically inflamed human intestine.

Cell-mediated lymphocyte cytotoxicity in ileum and colon of patients with ulcerative colitis (UC), Crohn's disease (CD) and controls was investigated. Frequencies of cells expressing perforin and Fas-ligand (FasL) were determined by immunomorphometry. mRNA expression of perforin, granzyme B and FasL in T cells and subsets was assayed by reverse transcriptase-polymerase chain reaction. Cytotoxicity of intraepithelial and lamina propria lymphocytes was analysed without ex vivo activation in three functional assays: (1) anti-CD3-dependent T-cell receptor (TCR)-/CD3-mediated redirected cytotoxicity, (2) Fas-/FasL-mediated TCR-/CD3-independent cytotoxicity and (3) natural killer (NK) cell cytotoxicity. Inflammation in ileum of CD patients caused increased frequency of perforin-expressing cells and enhanced perforin-dependent TCR-/CD3-mediated cytotoxicity. In contrast, lymphocytes in the inflamed colon of UC or Crohn's colitis patients did not display this cytotoxicity nor did lymphocytes of normal colon. Normal colon lymphocytes showed spontaneous Fas-/FasL-mediated cytotoxicity. This activity was retained but not enhanced in inflamed UC colon. In contrast, a significant increase of FasL-expressing cells was seen in situ. Inflammation did not induce NK cell activity in colonic lymphocytes. Intestinal lymphocytes comprise effectors active in two different cytolytic processes. 'Classical' cytotoxic T lymphocytes in small intestine and lymphocytes executing TCR-/CD3-independent FasL-/Fas-mediated killing of unknown biological role present throughout the intestinal mucosa. Ongoing normal cytolytic processes seem to be enhanced by chronic inflammation.

Adult↗
PubMed1975

An in vitro inhibitor of Na-K-ATPase present in an adenosinetriphosphate preparation.

An in vitro inhibitor of Na-K-ATPase was discovered in a commonly used preparation of ATP made by the Sigma Chemical Company, St. Louis, Mo. (Sigma grade ATP). As measured by a reliable and widely used assay system in which phosphate liberation in measured colorimetrically, Na-K-ATPase activity in the rat kidney, small intestine and colon was about 50% lower then Sigma grade ATP was used as substrate as compared to another Sigma Chemical Company product II ATP. Mg-ATPase and adenylate cyclase assays were unaffected by substituting Sigma grade for grade II ATP. The inhibitor of Na-K-ATPase could not be identified. Sigma grade ATP probably should not be used when measuring the activity of Na-K-ATPase in the rat kidney, small intestine, or colon.

Adenosine Triphosphatases↗
PubMed1994

recA mutations reduce adherence and colonization by classical and El Tor strains of Vibrio cholerae.

Two recA mutants of Vibrio cholerae (classical and El Tor biotypes) were constructed by disruption of the wild-type recA gene with mutated recA sequences of V. cholerae cloned in the suicide vector pGP704. Mutants defective in the recA gene were compared with their respective RecA+ parent strains with regard to their adherence to isolated rabbit intestine and colonization of intestine of infant mice. The recA mutation in V. cholerae was found to diminish adherence and markedly affected colonization.

Animals↗
PubMed2000

Intimin-mediated tissue specificity in enteropathogenic Escherichia coli interaction with human intestinal organ cultures.

The hallmark of enterohemorrhagic Escherichia coli (EHEC) and enteropathogenic E. coli (EPEC) adhesion to cultured human host cells is intimate attachment and the formation of attaching and effacing (A/E) lesions. Recently, EHEC O157:H7 was shown to induce A/E lesions on human intestinal explants. Unlike EPEC, which colonized the small intestine, EHEC adhesion was restricted to follicle-associated epithelium (FAE) of ileal Peyer's patches. This study tested the hypothesis that the bacterial adhesin intimin contributes to tissue specificity. Complementing the eae gene mutation in CVD206 (derived from EPEC strain E2348/69) with EPEC eaealpha (encoding intimin-alpha) restored the ability to colonize small intestinal mucosa like the parent strain. In contrast, complementing with EHEC eaegamma (encoding intimin-gamma) resulted in the strain adhering and inducing A/E lesion on Peyer's patches, similar to EHEC. An intimin-gamma-positive O55:H7 EPEC also targeted FAE. Thus, intimin contributes to the tissue specificity of A/E lesion-forming microbial pathogens.

Adhesins, Bacterial↗
PubMed1988

Potential impact of colonic irrigation on the indigenous intestinal microflora.

Under normal physiological conditions, the indigenous intestinal microflora is stable. The stability protects the host from colonization of nonindigenous microorganisms, including pathogens. Indigenous microorganisms capable of producing toxins are held in check by competing microflora, but any accidentally absorbed toxins are efficiently detoxified by the liver in the absence of hepatic pathology. Antibiotics are known to disrupt the normal flora leading to increased susceptibility to infection with other members of the normal flora or to toxins produced by them. It is possible that mechanical disruption of the flora by colonic irrigation could have a similar effect. Inadequately disinfected colonic irrigation machines have been the source of documented iatrogenic infection with Entamoeba histolytica. The potential for iatrogenic transmission of AIDS in a similar fashion must be considered.

Colon↗
PubMed2002

Ubiquitous production of macrophage migration inhibitory factor by human gastric and intestinal epithelium.

BACKGROUND & AIMS: Macrophage migration inhibitory factor (MIF) inhibits macrophage migration and has pleiotropic activities on immune and inflammatory responses, cell growth, and glucose metabolism. MIF is produced by T cells, macrophages, and endothelial cells. Because intestinal epithelial cells produce mediators important for regulating mucosal immune and inflammatory responses, we sought to determine if these cells produce MIF. METHODS: MIF expression was determined by immunostaining of human intestinal mucosa, intestinal xenografts, and cultured cells. MIF protein levels were quantitated by enzyme-linked immunosorbent assay and immunoblot analysis, messenger RNA was assessed by real-time reverse-transcription polymerase chain reaction, and functional activity was assessed by enzymatic and migration assays. RESULTS: MIF was abundantly expressed in vivo in gastric, small intestinal, and colonic epithelium and in epithelium lining human intestinal xenografts. MIF was also constitutively expressed at the messenger RNA and protein level by several cultured colon and gastric epithelial cell lines, and its expression in those cells was not up-regulated by the proinflammatory cytokines interleukin 1alpha, tumor necrosis factor alpha, or interferon gamma. Epithelial MIF from cultured cells was released predominantly from the apical side after Salmonella infection, had tautomerase activity, and arrested macrophage migration. CONCLUSIONS: Human intestinal epithelial cells are a major source of MIF, a molecule that can regulate macrophage migration, inflammation, and cell metabolism.

Animals↗
PubMed1990

Effect of dietary lipids on hepatic and intestinal monooxygenases in mice.

The effect of dietary lipids on hepatic and intestinal monooxygenases was studied by feeding C57BL/6N mice (for 2 wks) diets containing 5% and 23.5% (wt/wt) olive oil or corn oil. At the end of the feeding period, we measured arylhydrocarbon hydroxylase (AHH) activity in S9 preparations from liver, small intestine, and colon; and, using the same S9 preparations from the liver, we observed the activation of the following three dietary promutagens: 2-amino-3-methylimidazo(4,5-f)quinoline, 2-amino-3,8-dimethylimidazo(4,5-f) quinoxaline, and 2-amino-6-methyldipyrido(1,2-a:3',2'-d)imidazole. The results showed that high-fat diets increased hepatic AHH activity both in corn oil and olive oil diets compared with the low-fat diets; also, a 5% corn oil diet had significantly higher AHH activity compared with the 5% olive oil diet. AHH activity was, respectively, 48.6 +/- 5.1 and 79.5 +/- 11.4 pmol 3OH-benzo[a]pyrene formed/mg/min in the 5% and 23.5% olive oil diets and 66.1 +/- 5.1 and 83.9 +/- 12.2 in the 5% and 23.5% corn oil diets; values are means +/- SE, n = 16. The results also showed a significant increase in the ability of hepatic S9 fractions from animals on high-fat diets to activate promutagens in the Salmonella/plate test. On the contrary, AHH activity in the small intestine and colon was not affected by the fat content of the diet.

Animals↗
PubMed1996

Conserved expression of intestinal trefoil factor in the human colonic adenoma-carcinoma sequence.

Human intestinal trefoil factor (ITF), a mucosa-associated trefoil peptide, has been implicated in maintenance of mucosal integrity and may denote commitment to goblet cell differentiation. We have quantitated and localized ITF expression in normal and neoplastic colons, determined the molecular forms present, and examined the relationships among ITF expression and mucin production and increasing dysplasia. Normal and neoplastic human colonic mucosa (n = 30) were extracted for quantitation by ITF radioimmunoassay and size determination by gel filtration and immunoblotting. Paraffin sections of normal bowel, hyperplastic polyps, adenomatous polyps, and adenocarcinoma were examined for ITF immunohistochemistry and mucin histochemistry. The predominant molecular species in both normal and neoplastic colon was a 7-kd monomer. Staining was localized to goblet and Paneth cells and the luminal surface in normal colon and in most adenomatous polyps but not hyperplastic polyps; this colocalized with periodic acid-Schiff histochemistry. ITF staining of undifferentiated cells was seen with increasing dysplasia, and Golgi region immunoreactivity was highly conserved in adenocarcinoma, independent of the presence of periodic acid-Schiff-positive mucin. ITF concentration in colonic extracts was 10 to 200 pmol/g. Levels in normal (88.1 +/- 15.1 pmol/g) and malignant (90.1 +/- 12.7 pmol/g) tissue were comparable. In carcinomas, there were significant associations among ITF expression and degree of differentiation and mucin presence. Loss of expression was associated with tumor necrosis and advanced Duke's stage. ITF is uniformly processed in normal and neoplastic colons. Goblet cell-derived ITF is associated with stainable mucin production. ITF synthesis by non-goblet colonocytes, however, is highly conserved in neoplastic differentiation.

Adenoma↗
PubMed1987

Evidence for regulation of human colonic mucosal immunoglobulin secretion by intestinal lymphoid cells.

In vitro immunoglobulin secretion has been studied using isolated colonic mucosal lymphoid cell populations obtained from 41 patients with non-inflammatory intestinal disease. Cell fractions were separated into intra-epithelial and lamina propria-enriched populations. The secretion of immunoglobulins by mucosal cells appeared to be independent of mitogen stimulation. When intra-epithelial lymphocyte preparations were co-cultured with autologous lamina propria lymphocytes, the secretion of IgM was significantly depressed but that of IgA and IgG was preserved. Co-cultured of mucosal cells with autologous peripheral blood lymphocytes resulted in suppression of pokeweed mitogen stimulated and unstimulated immunoglobulin secretion. Lamina propria T cells were shown to provide helper function for the in vitro secretion of IgA and IgM but not IgG, by autologous peripheral blood B cells. Immunoglobulin secretion by lamina propria lymphocytes was shown to be partly dependent on the concentration of E-rosette forming cells. These experiments demonstrate that human colonic mucosal lymphoid cells contain specific populations of helper and suppressor cells which selectively control intestinal immunoglobulin secretion.

Biopsy↗
PubMed1998

Evaluation of intestinal pressure-controlled colon delivery capsule containing caffeine as a model drug in human volunteers.

The delivery ability of a pressure-controlled colon delivery capsule (PCDC) containing caffeine as a test drug was evaluated after oral administration to healthy male human volunteers. The driving force causing PCDC disintegration in the intestinal tract is the physiological luminal pressure which results from peristalsis. Three kinds of PCDCs having different thickness of a water-insoluble polymer membrane was prepared by coating the inner surface of the gelatin capsules with ethylcellulose (EC). The mean thickness were 40 +/- 1 (SE) for type 1, 44 +/- 1 for type 2 and 50 +/- 1 micron for type 3 PCDC, respectively. Caffeine was dissolved with a suppository base (PEGs 400 and 1000) and the capsules were filled. Doses were 15, 45 or 75 mg. After blank saliva samples were obtained, test preparations were orally administered to the volunteers and saliva samples were collected for 1 min intervals hourly from 1 to 10 h in the fasted state study, and from 1 to 20 h and at 25 h in the fed state study. Caffeine concentrations in the saliva samples were analyzed by HPLC. The maximum salivary caffeine excretion rate increased as the oral caffeine dose increased. The maximum salivary caffeine excretion rate increased predominantly compared to the pre-dose level in 75 mg dose study. Therefore, all following studies were performed with this dose. The first appearance time of caffeine into the saliva, TI, was used as a parameter to estimate the disintegration time of test preparations in the gastrointestinal tract. The mean TI of types 1, 2, and 3 PCDCs were 3.0 +/- 0.4, 4.0 +/- 0.4 and 4.5 +/- 0.3 h, respectively. After oral administration of 75 mg caffeine in pain gelatin capsule as a reference preparation, caffeine appeared in the saliva within 0.5 h. The mean hardness of the PCDCs were 1.05 +/- 0.10 (type 1), 1.55 +/- 0.06 (type 2) and 2.08 +/- 0.15 newton (type 3), respectively. There were good correlations between three parameters: EC coating membrane thickness, hardness and TI (determination coefficient r2 = 0.935 between TI and thickness, r2 = 0.998 between thickness and hardness, r2 = 0.958 between hardness and TI). The effect of food intake on the delivery ability was examined with type 3 PCDCs. Food intake prolonged the mean TI, from 4.5 +/- 0.3 to 7.8 +/- 1.3 h. This increase is thought to be ascribed to prolonged gastric emptying time. Comparison with reported colon arrival times indicates that the type 3 PCDC functions in colon delivery of caffeine and is thought to be applicable to other drugs.

Adult↗
PubMed2001

Evaluation of avian-specific probiotic and Salmonella enteritidis-, Salmonella typhimurium-, and Salmonella heidelberg-specific antibodies on cecal colonization and organ invasion of Salmonella enteritidis in broilers.

Salmonella Enteritidis colonizes the intestinal tract of poultry and causes foodborne illness in humans. Reduction of Salmonella Enteritidis colonization in the intestinal tract of poultry reduces potential carcass contamination during slaughter. The purpose of this study was to determine the effect of an avian-specific probiotic combined with Salmonella Enteritidis-, Salmonella Typhimurium-, and Salmonella Heidelberg-specific antibodies on the cecal colonization and organ invasion of Salmonella Enteritidis in broiler as well as on body weights. The treatment group was defined as chicks spray-vaccinated with Avian Pac Plus at the hatchery and given Avian Pac Plus for the first 3 days after placement. An intermediate treatment was given at 10 and 14 days, 2 days prior to vaccination and 2 days postvaccination. All birds were vaccinated with Newcastle disease vaccine, La Sota virus (one drop/eye) at 12 days of age. A final treatment was given 3 days preslaughter. The control group was defined as chicks not given Avian Pac Plus at any time. Six hours after oral administration of the probiotic suspension (treatment group) or water (control group) at placement, the chicks were challenged with Salmonella Enteritidis. All chickens were orally inoculated with 0.25 ml of Salmonella Enteritidis that contained 4 x 10(7) CFU/1.0 ml. Cecal colonization and organ invasion were evaluated for Salmonella Enteritidis on days 0, 1, 3, 7, 10, 17, 24, 31, 38, and 41. The probiotic-treated group had a significantly lower concentration of Salmonella Enteritidis cecal colonization at days 3, 7, 10, 17, 24, 31, 38, and 41 when compared to the nontreated, control group (P < 0.05). Similarly, there was a significant difference (P < 0.05) in the isolation of Salmonella Enteritidis from the internal organs (liver and spleen) when probiotic-treated and nonprobiotic-treated groups were compared. There was no significant difference (P > 0.05) in the mean body weight between the two experimental groups at each collection period. These results indicated that a combination of Lactobacillus acidophilus, Streptococcus faecium, and Salmonella Enteritidis, Salmonella Typhimurium, and Salmonella Heidelberg-Specific antibodies have a beneficial effect in reducing the colonization of Salmonella Enteritidis in market-aged broilers.

Animals↗
PubMed1995

Co-localization of NADPH diaphorase reactivity and vasoactive intestinal polypeptide in human colon.

Nitric oxide-containing nervous structures were localized in the human colon using NADPH diaphorase activity and nitric oxide synthase immunoreactivity. We found some, solitary NADPH diaphorase-reactive and nitric oxide synthase-immunoreactive neurons in the submucous plexus, while the myenteric plexus contained several neurons, often arranged in clusters, and nerve fibers showing these markers. The circular muscle layer contained a dense plexus of NADPH diaphorase-reactive nerves, which was greater than that in the longitudinal muscle layer. We report on co-localization of NADPH diaphorase activity and VIP immunoreactivity in several neurons of the myenteric ganglia. Such co-localization has not been reported previously for human colon. Localization of nitric oxide synthase and VIP in the myenteric plexus and in the nerves of circular muscle layer raises the possibility that nitric oxide contributes to the regulation of motility in the human colon.

Colectomy↗