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Bacterial colonization of the large intestine in newborns delivered by cesarean section.

Colonization of the gastrointestinal tract in newborns delivered by Cesarean section occurs during the first days of life by bacteria provided by the environment. It is more rapid in breast-fed infants than in bottle-fed babies. The intestinal flora is more diversified if the infants receive formula feeding. The first bacteria encountered are facultative anaerobes; they remain predominant during the first two weeks of life. In comparison to vaginal delivery, there were low levels of strict anaerobes after Cesarean section; members of the Bacteroides fragilis group were still completely lacking after 14 days of life and Bifidobacteria were only isolated sporadically.

Bacteria, Anaerobic↗

An approach to analyze mechanisms of intestinal adaptation following total proctocolectomy.

We hypothesized that epithelial cells of the remnant small intestine display "colonic" phenotype after total proctocolectomy. The aims of the present study were to identify preferentially expressed molecules in the colon or in the small intestine and to evaluate mRNA levels of those in the ileal pouch. Differential gene expression was investigated between the small intestine and the colon by using cDNA microarray and was confirmed by Northern blotting. Expression of three colonic mRNAs (3-hydroxy-3-methylglutaryl-coenzyme A synthase 2, deleted malignant brain tumors 1, carcinoembryonic antigen-related cell adhesion molecule 1) and one "small intestinal" (microsomal triglyceride transfer protein) mRNA were compared between the control and the ileal pouch mucosae by quantitative reverse transcriptase-polymerase chain reaction. Seventy-four clones were differentially expressed with more than a threefold difference. Differential expression was confirmed in all mRNAs examined, including 3-hydroxy-3-methylglutaryl-coenzyme A synthase 2 and microsomal triglyceride transfer protein. The mucosal expression of carcinoembryonic antigen-related cell adhesion molecule 1 mRNA in the ileal pouch was enhanced in humans. The remnant ileum develops some, but not all, colonic phenotype after total proctocolectomy. Comparative study of epithelial gene expression between the small intestine and the colon enables us to analyze mechanisms of intestinal adaptation after total proctocolectomy.

Adaptation, Physiological↗

Molecular cloning and characterization of a novel UDP-GlcNAc:GalNAc-peptide beta1,3-N-acetylglucosaminyltransferase (beta 3Gn-T6), an enzyme synthesizing the core 3 structure of O-glycans.

The core 3 structure of the O-glycan, GlcNAcbeta1-3GalNAcalpha1-serine/threonine, an important precursor in the biosynthesis of mucin-type glycoproteins, is synthesized by UDP-N-acetylglucosamine:GalNAc-peptide beta1,3-N- acetylglucosaminyltransferase (beta3Gn-T; core 3 synthase). The core 3 structure is restricted in its occurrence to mucins from specific tissues such as the stomach, small intestine, and colon. A partial sequence encoding a novel member of the human beta3Gn-T family was found in one of the data bases. We cloned a complementary DNA of this gene and named it beta3Gn-T6. The putative amino acid sequence of beta3Gn-T6 retains the beta3Gn-T motifs and is predicted to comprise a typical type II membrane protein. The soluble form of beta3Gn-T6 expressed in insect cells showed beta3Gn-T activity toward GalNAcalpha-p-nitrophenyl and GalNAcalpha1-serine/threonine. The beta1,3-linkage between GlcNAc and GalNAc of the enzyme reaction product was confirmed by high performance liquid chromatography and NMR analyses. beta3Gn-T6 effectively transferred a GlcNAc to the GalNAc residue on MUC1 mucin, resulting in the synthesis of a core 3 structure. Real time PCR analysis revealed that the beta3Gn-T6 transcript was restricted in its distribution, mainly to the stomach, colon, and small intestine. We concluded that beta3Gn-T6 is the most logical candidate for the core 3 synthase, which plays an important role in the synthesis of mucin-type O-glycans in digestive organs.

Amino Acid Sequence↗

Bifidobacterium longum, a lactic acid-producing intestinal bacterium inhibits colon cancer and modulates the intermediate biomarkers of colon carcinogenesis.

The human colon can be described as a complex microbial ecosystem, comprising several hundred bacterial species. Some of these enteric bacteria are beneficial to the host and have been shown to exert antimutagenic and anticarcinogenic properties. We have investigated the colon tumor inhibitory activity of Bifidobacterium longum, a lactic acid-producing enterobacterium. The modifying effects of this lactic culture on colonic mucosal and/or tumor cell proliferation, ODC activity and ras-p21 oncoprotein expression in colon carcinogenesis were also analyzed. Male F344 rats were fed a modified AIN-76A diet containing 0 or 2% lyophilized cultures of B. longum and s.c. administered azoxymethane (AOM) dissolved in normal saline at a dose of 15 mg/kg body wt, once weekly for 2 weeks. Vehicle controls received an equal volume of normal saline s.c. Animals were maintained on control or experimental diets until termination of the study. Animals intended for analysis of cell proliferation were killed 20 weeks after the second AOM injection, whereas animals intended for colon tumor analysis and measurement of ODC activity and ras-p21 expression were killed 40 weeks after the last AOM injection. The data demonstrate that dietary administration of lyophilized cultures of B. longum resulted in significant suppression of colon tumor incidence and tumor multiplicity and also reduced tumor volume. Results also revealed that ingestion of B. longum significantly inhibited AOM-induced cell proliferation, ODC activity and expression of ras-p21 oncoprotein. Data suggest that oral administration of probiotic B. longum exerts strong antitumor activity, as indicated by modulation of the intermediate biomarkers of colon cancer, and consequently reduced tumor outcome.

Animals↗

An animal model of Mycobacterium avium complex disseminated infection after colonization of the intestinal tract.

Mycobacterium avium complex infections occur in 30%-80% of patients with AIDS. Recent evidence supports the gastrointestinal tract as the source of M. avium. Although a reproducible animal model exists, a model more closely resembling the infection in AIDS patients is needed to answer pertinent questions regarding response to therapy and prophylaxis. Beige mice were infected orally (1 x 10(8) or 1 x 10(4) cfu, five doses), and consistent, reproducible disseminated infections after 4 and 8 weeks, respectively, were obtained. Bacteremia was observed in none to 70% of the animals depending on the strain used, and mortality ranged from none to 33%, also depending on the strain used. Concomitant ingestion of ethanol (4% of daily dietary calories) was associated with a significant increase in the number of viable bacteria recovered from liver, spleen, and appendix compared with animals not receiving ethanol. The orally infected animal model closely resembles M. avium infection in humans and may be important in investigating prophylaxis and therapy of this infection.

Acquired Immunodeficiency Syndrome↗

Factors controlling the bacterial colonization of the intestine in breastfed infants.

This article summarizes the published data on the intestinal microflora in breastfed infants published during the last 15 y. Enterobacteria and enterococci are found in high numbers in most infants during the first week of life. Bifidobacteria and Bacteroides spp. are found in increasing numbers at the following weeks. The intestinal microflora in breastfed infants can also be followed by different biochemical parameters. Acetic acid is found in higher concentrations in breastfed than in formula-fed infants. Degradation of mucin starts later in breastfed than in formula-fed infants. The conversion of cholesterol to coprostanol is also delayed by breastfeeding. Geographical differences in the composition of the intestinal microflora in infants have been reported, i.e. enterobacteria, enterococci, bifidobacteria, lactobacilli and bacteroides show different occurrences in developed and developing countries. There are minor differences in the infant's intestinal microflora due to breastfeeding or/and formula feeding.

Bacteria, Aerobic↗

Colonization of the intestinal tract of conventional mice with Candida albicans and treatment with antifungal agents.

Conventional mice inoculated with Candida albicans per os were unable to maintain this organism in the intestinal tract as judged by decreasing numbers of yeast recoverable from feces. After inoculation with 10(7) cells/mouse, fecal counts ranged from 10(5) cells per g of feces to 5 x 10(3) cells per g of feces during a 12-day experimental period. Addition of various antibiotics to the drinking water did not result in any improvement in maintenance or stability of the gut population. A combination of X irradiation and administration of tobramycin or gentamicin, however, resulted in a stable population of C. albicans in the intestinal tract, with cell counts in the feces remaining constant at a level of about 10(6)/g of feces for a period of 10 to 15 days. The usefulness of this model in assessing the effect of experimental drugs on C. albicans infections of the gut was demonstrated by the fact that treatment with a new antifungal antibiotic (A9145), amphotericin B, 5-fluorocytosine, or nystatin resulted in a reduction in the fecal counts of C. albicans from experimentally infected animals.

Animals↗

Enteric beta-defensin: molecular cloning and characterization of a gene with inducible intestinal epithelial cell expression associated with Cryptosporidium parvum infection.

A growing body of evidence suggests that endogenous antibiotics contribute to the innate defense of mammalian mucosal surfaces. In the cow, beta-defensins constitute a large family of antibiotic peptides whose members have been previously isolated from the respiratory and oral mucosa, as well as circulating phagocytic cells. A novel bovine genomic clone with beta-defensin-related sequence [corrected] related to those of these alpha-defensins was isolated and characterized. The corresponding cDNA was isolated from a small intestinal library; its open reading frame predicts a deduced sequence of a novel beta-defensin, which we designate enteric beta-defensin (EBD). Northern blot analysis of a variety of bovine tissues revealed that EBD mRNA is highly expressed in the distal small intestine and colon, anatomic locations distinct from those for previously characterized beta-defensins. EBD mRNA was further localized by in situ hybridization to epithelial cells of the colon and small intestinal crypts. Infection of two calves with the intestinal parasite Cryptosporidium parvum induced 5- and 10-fold increases above control levels of EBD mRNA in intestinal tissues. An anchored-PCR strategy was used to identify other beta-defensin mRNAs expressed in the intestine. In addition to that of EBD, several low-abundance cDNAs which corresponded to other beta-defensin mRNAs were cloned. Most of these clones encoded previously characterized beta-defensins or closely related isoforms, but two encoded a previously uncharacterized prepro-beta-defensin. Northern blot evidence supported that all of these other beta-defensin genes are expressed at levels lower than that of the EBD gene in enteric tissue. Furthermore, some of these beta-defensin mRNAs were abundant in bone marrow, suggesting that in enteric tissue their expression may be in cells of hematopoietic origin. Extracts of small intestinal mucosa obtained from healthy cows have numerous active chromatographic fractions as determined by an antibacterial assay, and one peptide was partially purified. The peptide corresponded to one of the low-abundance cDNAs. This study provides evidence of beta-defensin expression in enteric tissue and that the mRNA encoding a major beta-defensin of enteric tissue, EBD, is inducibly expressed in enteric epithelial cells. These findings support the proposal that beta-defensins may contribute to host defense of enteric mucosa.

Amino Acid Sequence↗

Structure, function and gene expression of epithelial mucins.

In this review the main characteristics, i.e., structure, function and gene expression, of the different mucins are discussed. Mucin-type molecules consist of a core protein moiety (apomucin) where a number of carbohydrate chains are attached to serines and threonines by glycosidic bonds. O-linked carbohydrates form up to 80% of the molecule and the length of the glucidic side chains varies from one to more than 20 residues. At least eight mucin-like genes have been isolated so far, and the main characteristic is the presence of a central domain composed of a variable number of "tandem repeats". The sequence homology of the central domain among the different members of the mucin-type family is limited, indicating that this internal domain is unique for each mucin. Thanks to the integrated results of genetic, immunological and biochemical studies, it is now possible to identify eight apomucin genes, namely MUC1, MUC2, MUC3, MUC4, MUC5AC, MUC5B, MUC6 and MUC7. MUC1 is the best characterized mucin and it is expressed on the apical surface of most polarized epithelial cells. The MUC1 gene has been cloned and sequenced. The MUC2 gene encodes a typical secretory gel-forming mucin which represents the predominant form in human intestinal and colon tissues. Another intestinal mucin is MUC3. The MUC4, MUC5AC and MUC5B genes have been isolated from a bronchial tissue cDNA library. The MUC4 and MUC5AC genes are mainly expressed in the respiratory tract, in gastric and reproductive mucosa, while MUC5B is highly detectable only in the bronchial glands. The MUC6 gene is expressed by gastric tissue and, recently, MUC7 has been cloned and sequenced using a salivary cDNA library.

Amino Acid Sequence↗

[Intestinal pneumatosis. Right colon cystic form with linear aspect on scan].

INTRODUCTION: Digestive pneumatosis corresponds to the presence of air in a wall of digestive tract. It is common to oppose primary cystic-type pneumatosis, involving the left colon, with linear-type pneumatosis that may affect the whole of the digestive tract. OBSERVATION: In a 41 year-old man presenting with Sharp's syndrome, abdominal pain occurred revealing an isolated right colon pneumatosis. Although abdominal tomodensitometry showed a strictly linear pneumatosis, colposcopy revealed voluminous cysts. CONCLUSION: With this case report, the limits of tomodensitometry in distinguishing linear from cystic forms are apparent. The importance of avoiding surgery is underlined. Our patient presented with a pneumo-peritonitis only requiring medical treatment since this disease generally regresses spontaneously.

Adult↗

[Adrenergic reactivity of the gastro-intestinal tract. II. Colon of the guinea pig and cecum of the mice].

A study has been made of the alpha- and beta-adrenoreceptors in the guinea pig colon and the mouse caecum using noradrenalina as agonist and alpha- and beta-adrenolytics separately or together. In the guinea pig colon alpha-adrenolytics used alone did not alter the action of noradrenalin: one beta-lytic (propranolol) used alone has slight non dose-dependent action. The alpha-lytics used together with beta-lytic strongly potential their effect. These results are interpreted as indicating that the preparation contains alpha-receptors as well as beta-receptors; activation of the alpha-receptors is observed only when the beta-mimetic effect of noradrenalin is inhibited. In the mouse caecum alpha-lytics prove devoid of effect either alone or when associated with beta-lytics; it is deduced therefore that this preparation contains only beta-receptors.

Animals↗

[The relationship between carcinoembryonic antigen (CEA) expression and histogenesis of gastric cancer (application of immunohistochemical and mucin histochemical techniques)].

The distribution of CEA and mucin in gastric specimens of 134 cases was examined by immunohistochemical and mucin histochemical techniques. The results showed that in 85.58% of the gastric cancers CEA was positive, including all mucinous adenocarcinomas, signet-ring cell carcinomas and papillary adenocarcinomas. In tubular adenocarcinoma, there was a tendency of increased expression of CEA with the degree of cell differentiation. The positive rate of CEA in intestinal type of gastric cancer was higher than that in gastric type and stem cell type. Intestinal metaplasia with colonic type sulphomucin had a higher positive rate than that without sulphomucin. The positive rate of CEA in cancers secreting sulfomucin was higher than that in cancers without sulfomucin. It suggested that gastric cancers expressing CEA was histogenetically related to colonic type intestinal metaplasia and cancers without CEA expression might be evolved from gastric proper epithelium.

Adenocarcinoma↗

Molecular cloning of putative members of the Na/H exchanger gene family. cDNA cloning, deduced amino acid sequence, and mRNA tissue expression of the rat Na/H exchanger NHE-1 and two structurally related proteins.

Biochemical and pharmacological data support the existence of multiple forms of the Na/H exchanger (NHE). Two isoforms, termed NHE-1 and NHE-2, have recently been isolated from rabbit ileal villus epithelial cells (Tse, C. M., Ma, A. I., Yang, V. W., Watson, A. J. M., Levine, S., Montrose, M. H., Potter, J., Sardet, C., Pouysségur, J., and Donowitz, M. (1991) EMBO J. 10, 1957-1967; Tse, C. M., Watson, A. J. M., Ma, A. I., Pouysségur, J., and Donowitz, M. (1991) Gastroenterology 100, A258). To identify additional molecular forms of the exchanger, rat brain, heart, kidney, stomach, and spleen cDNA libraries were screened for their presence using an NHE-1 cDNA probe under low stringency hybridization conditions. cDNAs encoding rat NHE-1 and two structurally related proteins, designated NHE-3 and NHE-4, have been isolated. Based on the deduced amino acid sequences, NHE-1, -3, and -4 are similar in size, having relative molecular masses of 91,506, 92,997, and 81,427, respectively. Overall, the proteins exhibit approximately 40% amino acid identity to each other and have similar hydropathy profiles, suggesting that they have the same transmembrane organization. The predicted N-terminal transmembrane regions of the three proteins, which span between 453 and 503 amino acids, exhibit the highest degree of identity (45-49%). In contrast, the C-terminal cytoplasmic regions, which span between 247 and 378 amino acids, exhibit very low amino acid identity (24-31%). Tissue distribution studies reveal that the NHE-1 mRNA is present at varying levels in all tissues examined, whereas NHE-3 and NHE-4 mRNAs exhibit a more limited distribution. NHE-3 mRNA is expressed at high levels in colon and small intestine, with significant levels also present in kidney and stomach. NHE-4 mRNA is most abundant in stomach, followed by intermediate levels in small intestine and colon and lesser amounts in kidney, brain, uterus, and skeletal muscle. These data suggest that the molecular basis for the functional diversity of the Na/H exchanger in mammals is based, at least in part, on expression of multiple members of a gene family.

Amino Acid Sequence↗

[Peculiarities of microbial colonization of the intestinal tract in newborns and pre-term infants in intensive care units].

The aim of the study was to compare the development of intestinal microflora in clinically healthy newborns, born by mothers with physiological pregnancy, and in small premature infants, who were treated in intensive care units (ICU) using various regimens of antibacterial therapy. The study revealed that the most frequent bacteria found in the intestinal tract of healthy infants at the and of neonatal period were bifidobacteria, enterobacteria, and coagulase-negative staphylococci and enterococci. Together with large quantity of autochtonous bacteria, the study revealed conditionally pathogenic microorganisms, such as klebsiella and coagulase-positive staphylococci at the end of neonatal period. The intestinal microflora of premature infants in ICU, treated with a combination of third generation cephalosporins and aminoglycosides from the first hours of life, was characterized by total absence of indigenous microflora, and prevalence of enterococci and staphylococci. The results show that the first stage of antibacterial therapy of preterm infants in ICU should be based upon the principles of selective decontamination.

Age Factors↗

Bacterial interactions in the intestine of the newborn delivered by cesarean section.

The purpose of this study was to clarify the role of the intestinal anaerobic bacteria colonizing the intestine of the newborn delivered by cesarean section. Control of the intestinal microecology is dependent on many factors including intestinal peristalsis, the intraluminal environment, and microbial interactions, that deter the overgrowth of pathogens populations. Numerous factors help achieve this normal balance. The effect of feeding seems to induce bacteriological changes.

Bacteria, Anaerobic↗