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Host gene expression in the colon of gnotobiotic interleukin-2-deficient mice colonized with commensal colitogenic or noncolitogenic bacterial strains: common patterns and bacteria strain specific signatures.

BACKGROUND: Specific pathogen-free (SPF), but not germfree (GF), interleukin (IL)-2-deficient (IL-2-/-) mice develop inflammatory bowel disease (IBD) at 10 to 15 weeks of age. Gnotobiotic IL-2-/- mice monocolonized with E. coli mpk develop IBD at 25 to 33 weeks of age but not B. vulgatus mpk, E. coli Nissle 1917, or mice cocolonized with both E. coli mpk and B. vulgatus. METHODS: To determine genes regulated by these commensal bacteria, host gene expression in the colon of 8-week-old IL-2-/- mice was compared by using microarrays and semiquantitative reverse-transcription polymerase chain reaction. Colonization with E. coli mpk/B. vulgatus or SPF microbiota altered the gene expression profile more profoundly than monocolonization with either B. vulgatus, E. coli mpk or E. coli Nissle indicating that the complexity of the gene expression pattern is influenced by the diversity of the microbiota. RESULTS: A small but distinct group of genes could be defined which might be associated with colitis development. Thus, 8 week old E. coli mpk IL-2-/- mice rone to colitis compared to E. coli Nissle, B. vulgatus and E. coli mpk/B. vulgatus IL-2-/- mice displayed a lower expression of the anti-inflammatory RegIII family genes such as RegIII[gamma] and pancreatitis associated protein (PAP) and peroxisome proliferator-activated receptor-[gamma] regulated genes such as adipsin and adiponectin. CONCLUSION: The increased expression of these genes in B. vulgatus colonized mice might be associated with prevention of E. coli mpk triggered colitis in E. coli mpkM/B. vulgatus IL-2-/- mice.

Adipocytes↗

Differential expression of carcinoembryonic antigen and nonspecific crossreacting antigen genes in human colon adenocarcinomas and normal colon mucosa.

The level of mRNA for carcinoembryonic antigen (CEA) and nonspecific crossreacting antigen (NCA) in human colon adenocarcinomas and normal colon mucosa was analyzed by Northern blot hybridization using as probes 32P-labeled CEA cDNA and synthetic oligodeoxyribonucleotides specific to CEA and NCA mRNA sequences. The major 3.5-kb mRNA and a minor 4.2-kb mRNA are shown to be CEA-specific and expressed in both tissues, albeit at slightly different degrees, suggesting that the expression of CEA is regulated posttranscriptionally. Another minor mRNA of 2.9 kb is NCA-specific and expressed predominantly in cancerous tissues, suggesting its usefulness as a marker for colon cancer.

Adenocarcinoma↗

Acidification of the colonic mucins following polyvalent colonization of the germ-free rat.

Colonic mucins of germ-free (GF) and conventional rats (CV) were compared. After isolation by gel filtration on Sepharose CL-4B and purification by density gradient centrifugation, the content of isolated colonic mucins was estimated by determination of PAS positive carbohydrates. Purified mucins were subjected to carbohydrate and amino acid analysis and separated into mucin subclasses by ion exchange chromatography. While the total amount of colonic mucins was not statistically different in GF and CV animals, analysis of carbohydrate composition demonstrated an increased amount of sialic acid in CV rat mucin. This was in accordance with results of ion exchange chromatography, revealing a significant higher amount of negative charged mucin subclasses in CV mucin, compared to the germ-free counterpart. The results of amino acid analysis were similar in both groups. The compositional differences in carbohydrate moieties are attributed to modulations by the intestinal flora. A selective bacterial degradation of the neutral mucin subclasses and modifications in the mucin composition due to a stimulated synthesis are discussed.

Amino Acids↗

Relationship between the mouse colonizing ability of a human fecal Escherichia coli strain and its ability to bind a specific mouse colonic mucous gel protein.

Escherichia coli F-18, isolated from the feces of a healthy human, is an excellent colonizer of the large intestines of streptomycin-treated CD-1 mice. E. coli F-18 Col-, a poor mouse colonizer relative to F-18, lacks a 3 X 10(7)-dalton plasmid present in E. coli F-18. Both strains are human type A erythrocyte hemagglutination negative, have identical surface hydrophobicities, contain the same number of lipopolysaccharide molecules with the same O-side chain length, and have identical amounts of capsule. Differences between the two strains were observed. The relative amounts of specific outer membrane proteins differed, and E. coli F-18 was less motile than E. coli F-18 Col-. The abilities of the two strains to bind mouse large intestine mucous gel was also examined. Although each strain was able to use mucous gel as the sole source of carbon and nitrogen with equal ability, E. coli F-18 bound between two and three times more mucous gel than did E. coli F-18 Col-. Most of the difference in mucous gel binding ability of the two strains was accounted for by the greater ability of E. coli F-18 lipopolysaccharide to bind a single 26,000-dalton mucous gel protein. E. coli J5-3, a typical K-12 strain that is also a poor colonizer relative to E. coli F-18, was identical to E. coli F-18 Col- with respect to mucous gel binding ability.

Animals↗

Extensive colonization of the porcine colonic epithelium by a spirochete similar to Treponema innocens.

Specimens of colonic mucosa from two pigs with diarrhea were examined by light and electron microscopy. The epithelial surfaces of both pigs were extensively colonized by large spirochetes morphologically compatible with Treponema hyodysenteriae or Treponema innocens. The microorganisms were intimately attached end-on to the luminal cells. A weakly beta-hemolytic spirochete similar to T. innocens was isolated from the colon of one of the pigs.

Animals↗

Factors influencing the implantation of colon cancer cells on the colonic suture line in rats.

An experimental model is proposed to study suture line recurrence. By giving enemas with K12 colon carcinoma cells, derived from a 1,2-dimethylhydrazine-induced colon cancer in male syngeneic BDIX rats, we were able to obtain a suture implantation in about 30% of the control animals. Possible systemic and local factors which might influence this implantation rate could be studied using this model. Only local irrigations of the colon lumen with sodium hypochlorite 0.2% prior to anastomosis produced a significant decrease in implantation rate. Irrigations with chlorhexidine-cetrimide 2.5%, the use of iodized suture material and altering the cellular immune system of the hosts by ciclosporin or excessive surgical stress, had no significant effect on the implantation rate.

Animals↗

Activated CD4+ and CD8+ cells in the colonic mucosa of ulcerative colitis patients: their relationship to HLA-DR antigen expression on the colonic epithelium and serum soluble CD25 levels.

This study was performed to clarify the relationship between activated (HLA-DR-expressing) CD4+ and CD8+ cells in the colonic lamina propria of ulcerative colitis and other immunological factors, i.e., epithelial DR expression, serum soluble CD25 levels, and colonic mucosal CD25+ cells. The frequency of epithelial DR expression was positively correlated with the numbers of CD4+ and CD8+ cells. The percentages activated CD4+/CD4+ cells were higher in mucosae with DR- epithelium than in mucosae with DR+ epithelium. The serum soluble CD25 levels were increased in ulcerative colitis, and there was an inverse correlation between these levels and the relative number of activated CD4+ cells in untreated active disease. These results suggest that interactions among mucosal CD4+ cells, colonic epithelium, and serum soluble CD25 might play an important role in the pathogenesis of ulcerative colitis.

Adolescent↗

Colon-specific delivery of budesonide from microencapsulated cellulosic cores: evaluation of the efficacy against colonic inflammation in rats.

Budesonide (BDS) is a potent corticosteroid that has important implications in the pharmacotherapy of inflammatory bowel disease, especially in the treatment of ulcerative colitis and Crohn's disease. BDS is available on the market in the form of enteric-coated preparations. However these products, similar to other available site-specific dosage forms, are not sufficiently selective to treat colonic inflammatory bowel disease. The objective of this study was to evaluate the efficacy of a new microparticulate system containing BDS, to treat experimentally induced colitis in rats. This microparticulate system consisted of BDS-containing hydrophobic cores, microencapsulated within an enteric polymer, which solubilizes at above pH 7, thus combining pH-sensitive and controlled-release properties. Colonic injury and inflammation were assessed by measuring colon/bodyweight ratio, myeloperoxidase (MPO) activity, and by scoring macroscopic and histological damage in colitic rats. Rats were treated orally with BDS, included in the developed system, once a day for 4 days after the induction of inflammation. A BDS suspension and BDS-containing enteric microparticles were included as control formulations in the experimental design. The administration of the new BDS delivery system significantly reduced the colon/bodyweight ratio compared with the administration of control formulations. Similarly, MPO activity and macroscopic and histological damage of the inflamed colonic segments decreased significantly when the BDS formulation was administered, compared with the results obtained after oral administration of the drug suspension. There were no significant differences, however, when the new treatment was compared with the control formulation consisting of simple enteric microparticles.

Administration, Oral↗

The Shiga toxin 2 B subunit inhibits net fluid absorption in human colon and elicits fluid accumulation in rat colon loops.

Shiga toxin (Stx)-producing Escherichia coli (STEC) colonizes the large intestine causing a spectrum of disorders, including watery diarrhea, bloody diarrhea (hemorrhagic colitis), and hemolytic-uremic syndrome. It is estimated that hemolytic-uremic syndrome is the most common cause of acute renal failure in infants in Argentina. Stx is a multimeric toxin composed of one A subunit and five B subunits. In this study we demonstrate that the Stx2 B subunit inhibits the water absorption (Jw) across the human and rat colonic mucosa without altering the electrical parameters measured as transepithelial potential difference and short circuit current. The time-course Jw inhibition by 400 ng/ml purified Stx2 B subunit was similar to that obtained using 12 ng/ml Stx2 holotoxin suggesting that both, A and B subunits of Stx2 contributed to inhibit the Jw. Moreover, non-hemorrhagic fluid accumulation was observed in rat colon loops after 16 h of treatment with 3 and 30 ng/ml Stx2 B subunit. These changes indicate that Stx2 B subunit induces fluid accumulation independently of A subunit activity by altering the usual balance of intestinal absorption and secretion toward net secretion. In conclusion, our results suggest that the Stx2 B subunit, which is non-toxic for Vero cells, may contribute to the watery diarrhea observed in STEC infection. Further studies will be necessary to determine whether the toxicity of Stx2 B subunit may have pathogenic consequences when it is used as a component in an acellular STEC vaccine or as a vector in cancer vaccines.

Adult↗

Pre-elastofibroma-like colonic polyp: another cause of colonic polyp.

We present a case of pre-elastofibroma-like lesion, a kind of elastic-producing fibrous tumor. The small colonic polyp, which was found in a 49-year-old asymptomatic man in association with a large colonic adenoma, showed submucosal nodular deposits of fine granular or fibrillar eosinophilic materials with interspersed fibroblastic cells. Elastic stain revealed these deposits to consist mainly of dark gray granular or partially fibrillar dense elastinophilic materials, most of which were digested with elastase. This stromal lesion somewhat resembled a pre-elastofibroma. Therefore, pre-elastofibroma-like lesions should be kept in mind as a possible origin of colonic polyp.

Colonic Polyps↗

Pathways of nodal metastasis in carcinomas of the cecum, ascending colon, and transverse colon: CT demonstration.

The distribution of regional lymph node metastases in carcinomas of the cecum, ascending colon, and transverse colon follows the vascular distribution in the ileocolic mesentery, ascending mesocolon, and transverse mesocolon. The location of these metastatic nodes can be recognized on CT scans when the anatomy of the vessels in the ileocolic mesentery and mesocolon is well understood. This knowledge is important in the preoperative staging of carcinomas of the colon for curative surgery and in the early detection of recurrent nodal disease after curative surgery.

Cecal Neoplasms↗

Transneuronal labeling from the rat distal colon: anatomic evidence for regulation of distal colon function by a pontine corticotropin-releasing factor system.

Neural circuits that are positioned to regulate rat distal colon function were identified by immunohistochemical detection of pseudorabies virus (PRV) and corticotropin-releasing factor (CRF). The distribution of PRV-immunoreactive neurons was examined in spinal cord and brain at increasing times (72-118 hours) after distal colon injection. At 72-80 hours, PRV-labeling was confined to the spinal cord, in the parasympathetic preganglionic column in the lumbosacral spinal cord and in the intermediolateral column of the thoracic spinal cord. At longer survival times (88 hours), PRV-immunolabeled neurons in the lumbosacral spinal cord were also distributed in superficial layers of the dorsal horn, the dorsal commissure, and around the central canal. Trans-synaptic labeling was identified in the medullary raphe nuclei, parapyramidal region, A5, Barrington's nucleus, A7, and the dorsal cap of the paraventricular nucleus of the hypothalamus after longer survival times (88-91 hours). Substantial labeling of the locus coeruleus, periaqueductal gray and forebrain regions occurred at later survival times (> or = 96 hours). In dual-labeled sections, CRF terminal labeling surrounded PRV-labeled neurons in the parasympathetic preganglionic column of the lumbosacral spinal cord. Additionally, many neurons in Barrington's nucleus, but not other CRF-containing nuclei, were double labeled for CRF and PRV. These results, taken with previous studies, support a convergence in transneuronal labeling from different pelvic viscera that may be related to coordination of overall pelvic visceral functions. Importantly, they provide an anatomic substrate for an impact of CRF from Barrington's nucleus in normal and pathophysiological functions of the distal colon.

Animals↗

[Laparoscopy-assisted colonic polypectomy or how to be helped by laparoscopy to prevent colectomy in benign colonic polyps considered to be unresectable by colonoscopy].

The aim of this study was to prospectively evaluate the role of laparoscopic surgery in all patients presenting with colonic polyps. From April 1994 to April 1996, 16 consecutive patients were treated. The laparoscopy starts the procedure, then a colonoscopy easily and rapidly locates the colonic lesion. Under laparoscopic supervision a new snare polypectomy is often possible. If polypectomy remains impossible, a mini-laparotomy is performed above the polyp and allows polypectomy by extra-peritoneal colotomy. In one case, the polyp was not found on colonoscopy. Colonoscopic polypectomy was possible in 6 cases (40%), and 9 mini-laparotomies were necessary (60%). No colectomy was performed. The final histologic result showed two Dukes A carcinomas justifying secondary wide colectomy. Laparoscopy-assisted polypectomy is a safe and efficient procedure, allows complete excision of polyps and may avoid a colonic resection.

Aged↗

[Urothelial tumor and colonic cancer in the context of a syndrome of hereditary predisposition to HNPCC colonic cancer].

The authors report the association of ureteric tumour and colon carcinomas in the context of hereditary predisposition to HNPCC colon cancer (hereditary non polyposis colon cancer). The recall the diagnostic criteria of HNPCC syndrome and emphasize the importance of guiding the clinical interview of patients with upper urinary tract tumours in order to detect a family history and the presence of gastrointestinal tumours.

Colonic Polyps↗

[Large villous adenoma of the colon in carcinomatous transformation: colonic resection after endoscopic excision].

Large colonic villous adenomas are benign neoplasms capable of malignant transformation with a higher frequency than other adenomas. Such transformation often requires surgical therapy after endoscopic resection. The aim of the present study was to establish the indications for surgery in a series of 13 cases of large colonic villous adenomas initially submitted to endoscopic resection. The patients (8 males and 5 females; mean age; 62 years) were observed over the period 1993-2000. All endoscopic resections were performed using the piecemeal technique. In 7 cases there were 2 endoscopic sessions and in one case 3; a single case of post-endoscopic bleeding was treated conservatively. In 5 cases, endoscopic resection was deemed not to be radical and these patients were submitted to surgical resection. Histology on the surgical specimens revealed 2 cases of carcinoma (T1 and T2, respectively), confirmation of colonic villous adenoma in 2 cases and the presence of inflammatory tissue in 1 case. Among the patients treated with endoscopic resection alone one death occurred at two years due to lung and systemic metastases probably due to the malignant adenoma. After a review of the literature and on the basis of their own experience, the authors stress the importance of a combined pathological and endoscopic approach to establish when surgery is required.

Adenoma, Villous↗

Extensive liver metastasis from human colon cancer in nude and scid mice after orthotopic onplantation of histologically-intact human colon carcinoma tissue.

Clinically-relevant animal models of human cancer are greatly needed for the study of human cancer biology and the development of new cancer therapeutics and diagnostics. We report here that by orthotopically transplanting histologically-intact human colon cancer to the colon of the immunodeficient nude and scid mouse mutants that extensive local growth and liver metastases occur consistently even after extensive in vivo orthotopic passage. We demonstrate that the liver metastases arise by hematogenous spread. The models described in this report for human colon cancer should prove useful for individual cancer patients as well as for basic and applied studies to develop improved treatment.

Animals↗

Cell surface sialoprotein alterations in metastatic murine colon cancer cell lines selected in an animal model for colon cancer metastasis.

Alterations in cell surface proteins and glycoproteins may play a key role in determining the metastatic behavior of tumor cells. The cell surface proteins of a series of related murine colon cancer cells selected in an animal model for colon cancer metastasis (R. S. Bresalier et al., Cancer Res., 47: 1398-1406, 1987) were therefore compared by a variety of biochemical methods. Lactoperoxidase-catalyzed iodination of cell surface proteins followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated quantitative and qualitative differences in the cell surface protein profiles of parental cell line 51B (low metastatic potential) and its metastatic derivatives 51B LiM 5 and 51B LiM 6. Labeling of sialic acid-containing proteins suggested that, in the case of at least four of these proteins (Mr 170,000, 120,000, 95,000, and 55,000), this represented an increase in radioactive labeling of sialoglycoproteins from the metastatic lines. Affinity chromatography of solubilized 125I-labeled cell membrane proteins revealed a 2- to 3-fold increase in wheat germ agglutinin and Sambucus nigra lectin binding associated with the metastatic lines, compared to the poorly metastatic parent. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of material eluted from these columns demonstrated enhancement of proteins from the metastatic cells corresponding in molecular weight to the previously identified major sialoglycoproteins. Neuraminidase-releasable membrane-associated sialic acid and sialyltransferase activities were 2- to 3-fold higher in the metastatic cell lines compared to the parental line. Liver colonization after intrasplenic injection of the various lines into syngeneic mice was dramatically reduced by prior removal of cell surface sialic acid. Immunohistochemical staining of primary and metastatic tumors formed after cecal injection of parental 51B suggested selective metastasis by wheat germ agglutinin-binding tumor cells. These results further support the concept that cell membrane sialylation is important in determining the metastatic potential of cancer cells.

Animals↗

[A clinical study of 76 cases in carcinoma of the colon and rectum with metastasis to the peri-aortic lymph nodes. With particular reference to the skip metastasis in carcinoma of the sigmoid colon to the peri-aortic lymph nodes].

The therapeutic result by tumorous site were evaluated in 76 cases of carcinoma of the colon and rectum with the confirmed presence of metastasis to the peri-aortic lymph nodes. The results revealed that significant good out-comes were noted in the relatively non-curative cases of carcinoma of the sigmoid colon, with particularly good prognosis being found in six cases with skip metastasis. There was a 26.1% (6/23) incidence of the skip metastasis in carcinoma of the sigmoid colon with the peri-aortic lymph nodes metastasis. We confirmed two lymphatic routes of the skip metastasis; one route was from the lymph nodes near the tumor to the para-aortic lymph nodes directly, and the other was from the lymph nodes near the tumor to the lymph nodes at the aortic diffract directly and finally to the para-aortic lymph nodes. In the all six cases, the peri-aortic lymph nodes metastasis occurred only in inferior site of the inferior mesenteric artery. The average number of the peri-aortic lymph nodes metastasis were as few as 1.7 in these cases. The prognosis was favorable with three of the six cases scoring a three-year survival and included a five-year survival case.

Adult↗