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Expression of receptors for verotoxin 1 from Escherichia coli O157 on bovine intestinal epithelium.

Human enterohaemorrhagic Escherichia coli (EHEC) infection most commonly arises, either directly or indirectly, from cattle, which act as a reservoir host for these bacteria. In man, EHEC disease can be severe, whereas EHEC do not normally cause disease in cattle. Verotoxins (VTs) are the main virulence factors in human disease but no role for VT has been ascribed in cattle; however, this study shows for the first time that VT receptor is expressed by the bovine intestinal tract. VT bound to crypt epithelial cells of the small (ileum and jejunum) and large (caecum and colon) intestine independently of the animals' age. VT also bound to discrete cell subsets in the bovine kidney and to submucosal lymphoid cells but not to vasculature. Analysis of tissues for isoforms of the VT receptor, Gb3, confirmed the presence of the receptor in the bovine intestinal epithelium and kidney. A distinct pattern of Gb3 receptor isoform mixtures was observed in the bovine kidney. This, together with the general absence of receptors on vasculature, could contribute to the apparent resistance of cattle to systemic effects of VT. Expression of Gb3 on the bovine intestinal epithelium, together with previously described effects, may affect EHEC colonisation in its reservoir hosts and hence the potential for distribution to man.

Animals↗

[Intestinal perforation caused by chicken bone mimicking perforated colonic diverticulitis].

Intestinal Perforation can occur in less than 1% of cases of ingestion of foreign bodies. Clinical suspicion is very important to make the diagnosis since many medical conditions can simulate this pathology. We present a case of a 63 years old man who swallowed a chicken bone that perforated the sigmoid colon and produced a clinical picture mimicking a perforated colonic diverticulitis. At surgery, the bone was removed without any evidence of abscess formation. A Hartmann procedure was performed and we reanastomosed it 3 month later. The patient followed an uneventful postoperative course. We demonstrated that early detection and opportune surgical treatment decrease the risk of developing complications such as abscess formation, intestinal fistula or obstruction.

Animals↗

Mouse model for colonization and disease caused by enterohemorrhagic Escherichia coli O157:H7.

Enterohemorrhagic Escherichia coli O157:H7 isolates produce Shiga-like toxins and carry a 60-megadalton plasmid which encodes an adhesin for Henle 407 intestinal cells. A streptomycin-treated mouse model was used to compare the intestinal colonizing capacity of E. coli O157:H7 strain 933 with that of its 60-megadalton plasmid-cured derivative, strain 933cu. When fed individually to mice, both 933 and 933cu maintained a stable number of organisms per gram of feces, and the greatest numbers of 933 or 933cu were isolated from cecal and proximal colonic epithelial cells. When 933 and 933cu were simultaneously fed to mice, 933cu was unable to maintain a stable level of colonization in about two-thirds of the mice tested. However, in one-third of the mice, the number of 933cu in feces began to increase rapidly until a stable level of co-colonization with 933 was attained. The isolate from these mice, 933cu-rev, was excreted in high numbers when fed alone to mice and was found on epithelial cells throughout the entire large bowel and distal small intestine. Moreover, 933cu-rev grew in mucus from all segments of the intestine and at higher levels than strain 933 or 933cu. Only mice fed strain 933cu-rev died. Histopathological studies confirmed that mice fed 933cu-rev died from bilateral renal cortical tubular necrosis consistent with toxic insult, perhaps due to Shiga-like toxins. The virulence of 933cu-rev may reflect its ability to grow well in mucus and colonize the small as well as large bowel.

Animals↗

Quinolones and colonization resistance in human volunteers.

The suppression of alimentary canal flora by the three quinolones nalidixic acid, ciprofloxacin and pefloxacin was investigated in fifteen volunteers. They received the three quinolone compounds in tablet form both uncoated and colon-coated. Escherichia coli suppression was poor under nalidixic acid, but complete under ciprofloxacin and pefloxacin for both administration forms. The indigenous anaerobic flora contributing to the control of aerobic Streptococcus faecalis and Candida albicans in the intestines ('colonization resistance') was not affected by nalidixic acid and pefloxacin, and only slightly by ciprofloxacin. Out of the three quinolone compounds, only colon-coated pefloxacin was associated with a considerable absorption rate at colonic level. Using these criteria of successful Escherichia coli clearing from the intestinal canal--left the indigenous flora more or less intact (in a 'selective' way)--and a good absorption rate, pefloxacin is found to be superior to ciprofloxacin and nalidixic acid. These results suggest that a colon-coated tablet with a low dose of pefloxacin is a promising administration form in the therapy of recurrent urinary tract infections and diarrhoeal diseases and in the prevention of gut colonization in immunocompromised hosts.

Adult↗

Live attenuated oral cholera vaccines.

Live, orally administered, attenuated vaccine strains of Vibrio cholerae have many theoretical advantages over killed vaccines. A single oral inoculation could result in intestinal colonization and rapid immune responses, obviating the need for repetitive dosing. Live V. cholerae organisms can also respond to the intestinal environment and immunological exposure to in vivo expressed bacterial products, which could result in improved immunological protection against wild-type V. cholerae infection. The concern remains that live oral cholera vaccines may be less effective among partially immune individuals in cholera endemic areas as pre-existing antibodies can inhibit live organisms and decrease colonization of the gut. A number of live oral cholera vaccines have been developed to protect against cholera caused by the classical and El Tor serotypes of V. cholerae O1, including CVD 103-HgR, Peru-15 and V. cholerae 638. A number of live oral cholera vaccines have also been similarly developed to protect against cholera caused by V. cholerae O139, including CVD 112 and Bengal-15. Live, orally administered, attenuated cholera vaccines are in various stages of development and evaluation.

Cholera↗

The bacterial colonization of the large bowel of pre-term low birth weight neonates.

The bacterial colonization of the large bowel of 11 pre-term, low birth weight neonates who were nourished by expressed breast milk was examined by culturing serial faecal samples and compared to that observed in eight breast-fed and seven formula-fed full-term neonates. Pre-term neonates were colonized by high counts of facultatively anaerobic bacteria from the first days of life while bifidobacteria colonized only six babies during the first week and appeared in only one baby before day 5. Bacteroides spp. and clostridia were isolated from seven and six pre-term babies respectively during week 1 and were first observed on day 2. The intestinal colonization of pre-term infants differed from that in full-term breast-fed infants in the high counts of facultatively anaerobic bacteria and late appearance of bifidobacteria, and from both groups of full-term infants in the early stable colonization by Bacteroides spp. It is postulated that the composition of the normal intestinal microflora of pre-term low birth weight babies contributes to their predisposition to neonatal necrotizing enterocolitis. Results are discussed in relation to the effect of infant feeding regimens on intestinal microbial populations.

Actinomycetaceae↗

Cloning, expression analysis, and chromosomal localization of a novel butyrophilin-like receptor.

We isolated a cDNA clone which shows a similarity with human butyrophilin from a human colon mucosa cDNA library. The cDNA is 1964 bases long, with one open reading frame encoding a protein of 433 amino acids. The deduced amino acid sequence shows an overall homology of 36.5% with the human butyrophilin protein. This gene is mainly expressed in small intestine, colon, testis, and leukocytes. The chromosomal location of the gene was determined on the chromosome 5q35 region by polymerase chain reaction-based analysis with both a human/rodent monochromosomal hybrid cell panel and a radiation hybrid mapping panel.

Amino Acid Sequence↗

Reduced enterobacterial and increased staphylococcal colonization of the infantile bowel: an effect of hygienic lifestyle?

The modern Western lifestyle may have altered the composition of the commensal microflora. Here, we investigated the first year's intestinal colonization pattern in 99 vaginally delivered Swedish infants and 17 delivered by cesarean section. Rectal swabs obtained at 3 d of age were cultured for aerobic bacteria and fecal samples obtained at 1, 2, 4, and 8 wk and at 6 and 12 mo of age were cultivated quantitatively for aerobic and anaerobic bacteria. Vaginally delivered infants more often had Escherichia coli compared with cesarean section-delivered infants, whereas the latter more frequently carried other enterobacteria, such as Klebsiella and Enterobacter. Independent of delivery mode, it took 2 mo until most infants were colonized by enterobacteria, traditionally the first colonizers. In contrast, coagulase-negative staphylococci colonized 99% of the infants from d 3 onwards. The poor adaptation of staphylococci to the gut was shown by declining population sizes after some weeks. Dominating anaerobes were initially bifidobacteria and clostridia, whereas Bacteroides initially colonized only 30% of vaginally delivered infants and increased very slowly in prevalence. Bacteroides colonization was delayed up to 1 y in cesarean section-delivered compared with vaginally delivered infants. Our results show that some "traditional" fecal bacteria are acquired late today especially in cesarean section-delivered infants, probably due to limited environmental circulation. In their absence, skin bacteria like staphylococci have become the first gut colonizers.

Anti-Bacterial Agents↗

Effects of oral Lactobacillus GG on enteric microflora in low-birth-weight neonates.

BACKGROUND: Colonization patterns, especially by anaerobic flora, may play an important role in neonatal gut function. Probiotics could affect disease risk either directly through colonization or indirectly by promoting changes in gut microbial ecology. METHODS: To study the ability of Lactobacillus GG(LGG) to colonize the neonatal gut and modify its microbial ecology, a prospective, randomized study was performed in 71 preterm infants of less than 2000 g birth weight. Infants less than 1500 g (24 treated, 15 control) received 10(9) LGG orally twice daily for 21 days. Those infants weighing 1500 to 1999 g (23 treated, 9 control) were treated for 8 days. Stools were collected before treatment and on day 7 to 8 (and day 14 and 21, in the infants weighing less than 1500 g) for quantitative aerobic and anaerobic cultures. RESULTS: Colonization with LGG occurred in 5 of 24 (21%) infants who weighed less than 1500 g versus 11 of 23 (47%) in larger infants. Colonization was limited to infants who were not on antibiotics within 7 days of treatment with LGG. There was a paucity of bacterial species at baseline, although larger infants had more bacterial species (1.59 +/- 0.13 (SEM) vs 1.11 +/- 0.12; P < 0.03) and higher mean log colony forming units (CFU) (8.79 +/- 0.43 vs 7.22 +/- 0.63; P < 0.05) compared with infants weighing less than 1500 g LGG. Treatment in infants weighing less than 1500 g resulted in a significant increase in species number by day 7, with further increases by day 21. This increase was mainly the result of increased Gram (+) and anaerobic species. No difference in species number was noted in controls. Mean log CFU of Gram (-) bacteria did not change in treated infants weighing less than 1500 g. However, Gram (+) mean log CFU showed a significant increase on day 21 (6.1 +/- 0.9) compared with day 0 (3.5 +/- 0.9) (P < 0.05). No significant changes in species number or quantitative counts were noted after LGG treatment in the infants weighing 1500 to 1999 g LGG was well tolerated in all infants. CONCLUSION: The neonatal response to a probiotic preparation is dependent on gestational and post-natal age and prior antibiotic exposure. Although LGG is a relatively poor colonizer in infants, especially those infants weighing less than 1500 g at birth, it does appear to affect neonatal intestinal colonization patterns.

Administration, Oral↗

[Problems of the so-called geriatric gastrointestinal diseases].

The significant increase in the number of people older than seventy forces the physician to be acquainted with both psychological and physical alterations induced by aging and to devote an ever increasing proportion of time for recognition and treatment os such alterations. In the medical sense, the biological and physiological age is more important than the chronological age. With increasing age there is--especially concerning the digestive tract and its accessory organs--a rise in the incidence of organic affections and a decline in the frequency of functional disorders. Besides it is wise to know, that the increasing age there is often a coexistence of multiple degenerative disorders and disease states, involving many body systems and organs. On the background of this recognition it is also important to know, that prognosis too varies with age because of the coexistence of individually prognosticated disease states and moreover to realize, that elderly patients do not tolerate invasive and prolonged surgical procedures. Structural or functional disturbances of the digestive organs by aging processes do not cause death per se, but can become one important factor; degenerative sclerotic vascular alterations bear relationship to the poorly contractile vasculature that brings up difficulties in the control of hemorrhagic gastroduodenal ulcers. Many gastrointestinal disorders in elderly patients occur with an equal frequency in younger patients, some are more common in the geriatric population; these include hiatal hernia, carcinoma of esophagus, stomach, pancreas, bile ducts and colon, intestinal obstruction (ileus) by neoplastic growth, gallstone ileus, external hernia and operative adhesions and especially diverticular diseases of the colon and its complications and ischemic colitis by mesenteric vascular occlusion. Cirrhosis of the liver is often diagnosed for the first time in the older age groups while acute viral hepatitis uses to run a cholestatic course and is therefore often misdiagnosed as mechanical obstruction. In general history is difficult to obtain, the response of the organism with temperature and white blood count to stress is often delayed and rigidity associated with an underlying inflammatory disease involving the peritoneum is often delayed and rigidity associated with an underlying inflammatory disease involving the peritoneum is often atypical. Because of this limited reaction to severe stress, early surgical intervention is imperative in the elderly patients.

Aged↗

Immunoglobulin E (IgE) and IgE-containing cells in human gastrointestinal fluids and tissues.

Human gastric, small intestinal, colonic and rectal mucosae were examined for IgE-containing cells by single- and double-antibody immunofluorescence techniques, and IgE in intesinal fluids was measured by a double-antibody radioimmunoassay. IgE-containing cells were identified in all tissue specimens and comprised about 2% of all immunoglobulin-containing cells. Although less numerous than cells containing IgA, IgM or IgG, they were remarkably numerous in relation to the concentration of IgE in serum (about 0-001% of total immunoglobulin). IgE immunocytes were significantly more numerous in stomach and proximal small bowel than in colon and rectum, and were very numerous at bases of gastric and duodenal peptic ulcers. Measurable IgE was found in seventy-eight of eighty-five (92%) intestinal fluids. Sucrose gradient ultracentrifugation analysis of four of the fluids revealed that the immunologically reactive IgE was largely in fractions corresponding to molecules of lower molecular weight than that of albumin, which suggests that IgE in gut contents is degraded by proteolytic enzymes. The presence of IgE-forming cells in gastrointestinal tissues, and IgE or a fragment of IgE in intestinal fluids, suggests that IgE antibodies are available for participation in local reaginic-type reactions in the gut.

Adult↗

[Fecal intestinal flora in patients with colon adenoma and colon cancer].

Fecal intestinal flora in patients with colon adenoma showed significant decrease of Bifidobacterium. A tendency of decrease of Bifidobacterium and increase of Clostridium-other was shown in adenomas with higher degree atypia, but no definite tendency could be found concerning its size. Compared with the control group, patients with colon cancer showed significant increase of Clostridium-other and decrease tendency of Bifidobacterium which was not relevant to its percentage of total circum ference involved by the cancer. From the above findings, it is suggested that tumor growth or malignant transformation has relation to Bifidobacterium and Clostridium-other.

Adenoma↗

Strong biotype and serotype cross-protective antibacterial and antitoxic immunity in rabbits after cholera infection.

We studied whether immunity evoked by infection with classical or El Tor V. cholerae 01 organisms in rabbits (the RITARD model) also gives protection against cholera caused by V. cholerae of heterologous biotype as well as serotype, and whether such protection is antibacterial and/or antitoxic. A primary infection with a classical Ogawa or El Tor Inaba strain resulted in intestinal colonization and diarrheal disease in a dose-related manner though the El Tor strain was more virulent. As few as 10(3) El Tor organisms gave disease in more than 90% of the rabbits as compared to 10(9) classical organisms (ED90); the El Tor strain also gave rise to diarrhea with earlier onset and of greater severity and longer duration. The primary infection induced strong protective immunity against later challenge with either the homologous or the heterologous strain in doses that corresponded to 1,000 x ED90. Protection was associated with marked inhibition of colonization, and when rabbits convalescing from cholera infection were challenged with graded doses of bacteria or purified toxin in ligated intestinal loops significant antibacterial as well as antitoxic immunity was evident. Titer rises in serum vibriocidal and anti-lipopolysaccharide antibodies were similar after infection with either strain, whilst antitoxin titer rises were more marked after El Tor infection. During infection V. cholerae 01 organisms seem to express protective antigens that stimulate immunity which extends across both biotype and serotype barriers.

Animals↗

Jejunal brush border microvillous alterations in Giardia muris-infected mice: role of T lymphocytes and interleukin-6.

Intestinal colonization with the protozoan Giardia causes diffuse brush border microvillous alterations and disaccharidase deficiencies, which in turn are responsible for intestinal malabsorption and maldigestion. The role of T cells and/or cytokines in the pathogenesis of Giardia-induced microvillous injury remains unclear. The aim of this study was to assess the role of T cells and interleukin-6 (IL-6) in the brush border pathophysiology of acute murine giardiasis in vivo. Athymic nude (nu(-)/nu(-)) CD-1 mice and isogenic immunocompetent (nu(+)/nu(+)) CD-1 mice (4 weeks old) received an axenic Giardia muris trophozoite inoculum or vehicle (control) via orogastric gavage. Weight gain and food intake were assessed daily. On day 6, segments of jejunum were assessed for parasite load, brush border ultrastructure, IL-6 content, maltase and sucrase activities, villus-crypt architecture, and intraepithelial lymphocyte (IEL) infiltration. Despite similar parasitic loads on day 6, infected immunocompetent animals, but not infected nude mice, showed a diffuse loss of brush border microvillous surface area, which was correlated with a significant reduction in maltase and sucrase activities and a decrease in jejunal IL-6 concentration. In both athymic control and infected mice, jejunal brush border surface area and disaccharidases were high, but levels of tissue IL-6 were low and comparable to the concentration measured in immunocompetent infected animals. In both immunocompetent and nude mice, infection caused a small but significant increase in the numbers of IELs. These findings suggest that the enterocyte brush border injury and malfunction seen in giardiasis is, at least in part, mediated by thymus-derived T lymphocytes and that suppressed jejunal IL-6 does not necessarily accompany microvillous shortening.

Animals↗

Tea as a potential chemopreventive agent in PhIP carcinogenesis: effects of green tea and black tea on PhIP-DNA adduct formation in female F-344 rats.

The heterocyclic amine 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is formed during the cooking of proteinaceous animal foods (meat, chicken, and fish). PhIP is a carcinogen in the Fischer 344 (F-344) rat; it induces mammary tumors in female rats and lymphomas and colon and prostate tumors in male rats. In F-344 rats, PhIP forms DNA adducts in various organs, including the target organs. Inhibition of PhIP-DNA adduct formation is likely to lead to inhibition of PhIP tumorigenicity. We have examined the chemopreventive properties of green tea and black tea in PhIP carcinogenesis by evaluating their effects on PhIP-DNA adduct formation in the female F-344 rat. Young adult animals were maintained on powdered AIN-76A diet while receiving regular drinking water or 2% (wt/vol) infusions of green tea or black tea for a total of six weeks. During Weeks 3, 4, and 5, all animals received PhIP by gavage (1 mg/kg/day). Three rats per group were euthanized on Days 1 and 8 after termination of PhIP exposure. DNA was isolated from a number of organs and analyzed for PhIP-DNA adducts by 32P-postlabeling assays. Compared with animals on regular drinking water, PhIP-DNA adduct formation was inhibited in small intestine, colon, liver, and mammary epithelial cells (MECs) of animals receiving green tea or black tea as the sole source of drinking fluid. Green tea inhibited adduct formation in colon, liver, and MECs (33.3-80.0%) on both days, but only on Day 8 (54.4%) in small intestine. Black tea inhibited adduct formation on both days in liver (71.4-80.0%), on Day 1 in colon (40.0%), and on Day 8 in small intestine (81.8%); it had no effect on MEC adducts. Neither green tea nor black tea had an effect on adduct levels in pancreas, lungs, white blood cells, heart, kidneys, spleen, cecum, or stomach. Similarly, these teas did not affect the rate of adduct removal (percent change from Day 1 to Day 8) in any organ. It is concluded that green tea and black tea are potential chemopreventive agents in PhIP-induced tumorigenesis in the F-344 rat.

Animals↗

Effect of resection of small intestine on the interaction of vasoactive intestinal peptide with rat colonic epithelial cells.

Vasoactive intestinal peptide (VIP) receptors and VIP-dependent cyclic AMP production were studied in rat colonic epithelial cells 3 days after a 60% resection of the small intestine. Basal cyclic AMP levels were similar in both control and resected animals. The potency, but not the efficiency, of the peptide on the stimulation of cyclic AMP production was diminished in cells from resected rats. Accordingly, the affinity of VIP receptors, but not the binding capacity, decreased as a consequence of the loss of a part of the small intestinal mucosa. These observations are consistent with the known inhibitory role of cyclic AMP on cell proliferation in colonic epithelium and other tissues and suggest a participation of VIP acting through the cyclic nucleotide in the compensatory hyperproliferative response of the colon following massive resection of the small intestine.

Animals↗

The combined effects of dietary fat, protein, and energy intake on azoxymethane-induced intestinal and renal carcinogenesis.

Two 3 x 3 factorial experiments were conducted to examine the effects of dietary protein (8, 16, and 32% of energy from casein) and dietary fat (12, 24, and 48% of energy from corn oil) on the initiation and promotion of azoxymethane-induced carcinogenesis in rats. For the initiation study, 33 weanling male Sprague-Dawley rats were randomized to each of nine diets fed ad libitum. Azoxymethane was administered s.c. between the fourth to sixth weeks of feeding, providing a total dose of 6 mg/100 g body weight. All rats were subsequently fed a common diet containing 16% energy from protein and 24% energy from fat for an additional 30 to 38 weeks. For the promotion study, all rats were fed a common diet containing 16% of energy from protein and 12% of energy from fat until the completion of azoxymethane administration, when 33 rats were randomized to each of nine diets varying in fat and protein content and fed these diets until sacrifice. Low-protein diets during the initiation phase were associated with increased risk of renal adenocarcinomas (P less than 0.001) and mesenchymal (P = 0.005) malignancies. No other statistically significant relationships were found between the levels of dietary fat or protein and the prevalence of malignant lesions of the small intestine, colon, or kidney in either the initiation or promotion study (although polypoid adenocarcinoma of the colon increased suggestively from 13 to 19 to 26% of rats with increasing dietary protein during initiation). Results of a multiple logistic regression analysis, combining both studies, showed that ad libitum energy intake was significantly associated with intestinal carcinogenesis. The odds of finding an intestinal adenocarcinoma increased by 6.2 +/- 2.6% (SE) for each additional kilocalorie of mean daily ad libitum intake (P = 0.014). The quintile of rats which consumed the least averaged 60 kcal/day, while the most voracious quintile averaged 74 kcal/day. This 14 kcal/day difference in mean ad libitum intake corresponded to more than a doubling (146% increase) of the odds of developing an intestinal adenocarcinoma. These studies suggest that ad libitum energy intake is a critical factor modulating experimental colon carcinogenesis.

Adenocarcinoma↗

The activity of halofuginone in immunosuppressed rats infected with Cryptosporidium parvum.

Halofuginone was evaluated for its activity against experimentally induced infection due to Cryptosporidium parvum in rats rendered immunosuppressed with dexamethasone. The drug's activity was dose-related and both prophylaxis and therapy reduced the rate and severity of infection in the small intestine and caecum. Prophylactic treatment reduced infection of the common bile duct, but therapeutic administration did not and neither form of treatment reduced the infection rate in the colon. Intestinal infection recurred at a level comparable to that of untreated controls when treatment was discontinued. Treatment with halofuginone may reduce the severity of acute cryptosporidiosis, but is less efficacious for chronic cryptosporidiosis involving the colon and extraintestinal tissues, a manifestation increasingly seen in the immunocompromised host.

Animals↗