Microscopic colitis in children with chronic diarrhea.

OBJECTIVE: The aim of the present study was to study microscopic colitis (MC) in children with special reference to its role in chronic diarrhea and changes in mucosal biopsies. METHODS: A total of 100 consecutive children ages 3 to 12 years, with nonbloody diarrhea (passage of ≥3 loose stools per day) of >12 weeks' duration were screened and 26 were enrolled in the study in which no specific etiology could be found and colonoscopy did not reveal any mucosal abnormality. Colonic biopsies were evaluated for the presence of lymphocytic colitis or collagenous colitis and those with the characteristic changes were defined to have MC (group A). Colonic biopsies from patients with MC were compared with biopsies from patients with chronic diarrhea but no evidence of MC (group B). One hundred children ages 3 to 12 years with bleeding per rectum were screened and colonic biopsies from 45 patients (group C) who had colonic mucosal changes but no vascular or polyp lesion were compared with patients with MC. RESULTS: Of the 26 patients with chronic diarrhea, MC was found in 5 (3 lymphocytic colitis and 2 collagenous colitis). Significantly higher polymorphonuclear infiltration was seen in group A as compared with group B (13.8 [5.4-20.6] vs 7.2 [0-19.6]; P = 0.03) or group C (13.8 [5.4-20.6] vs 4 [0-13.4]; P = 0.007). Intraepithelial lymphocytes (12 [4-32] vs 4 [0-24]; P = 0.008) and basement membrane thickening (3.5 [2.9-10.6] vs 2.5 [1.6-5.86]; P = 0.008) were also significantly higher in group A as compared with group C. CONCLUSIONS: MC was found to be present in children with nonbloody chronic diarrhea in children. Further multicentric studies may provide adequate data on its prevalence.

Effects of zinc supplementation as adjunct therapy on the systemic immune responses in shigellosis.

BACKGROUND: Zinc is lost during diarrheal diseases, and zinc deficiency induces intestinal morphology-altering inflammatory responses that zinc supplementation can correct. OBJECTIVE: We assessed the in vivo effect of zinc supplementation on systemic and mucosal responses in mildly to moderately malnourished (defined as <-1 but >-2 and <-2 but >-3 weight-for-height z scores, respectively, based on the National Center for Health Statistics growth reference) children with shigellosis. DESIGN: A double-blind placebo-controlled trial was conducted in Shigella flexneri-infected children aged 12-59 mo. Daily for 14 d, elemental zinc (20 mg) and multivitamins (vitamins A and D, thiamine, riboflavin, and nicotinamide) plus calcium were given at twice the US recommended dietary allowance to the zinc group (n=28), and multivitamins plus calcium were given to the control group (n=28). All subjects received standard antibiotic therapy. RESULTS: There was no significant interaction between zinc supplementation and time, but zinc supplementation showed a significant effect on serum zinc concentrations. With a >or=4-fold increase in serum shigellacidal antibody titers from baseline used as the cutoff, the proportion of children with shigellacidal antibody response was greater in the zinc group than in the control group (P<0.03). There was a significant (P=0.02) treatment x time interaction for the proportions of circulating CD20+ and CD20+CD38+ cells, which were higher on day 7 in the zinc group than in the control group (P<0.007). No effect was seen on histopathologic features or the expression of innate and inflammatory mediators in the rectum. CONCLUSION: Adjunct therapy with zinc during acute shigellosis significantly improved seroconversion to shigellacidal antibody response and increased the proportions of circulating B lymphocytes and plasma cells.

Cytoskeletal changes during poliovirus infection in an intestinal cell line.

BACKGROUND & OBJECTIVES: Although polioviral replication has been extensively studied, cytoskeletal changes in the host cell during poliovirus replication have not been extensively investigated. We studied the ultrastructural and cytoskeletal changes in host cells during poliovirus infection. METHODS: Fluorescence staining of filamentous actin with a fluorescein-isothiocynate labelled mycotoxin, in the absence and presence of microfilament inhibitors cytochalasins B and D, and electron microscopy were used to investigate the role and fate of actin microfilaments during poliovirus infection, morphogenesis and release in an intestinal cell line, HRT-18. RESULTS: At 10 h post-infection, fluorescence staining of actin showed focal areas of fluorescence in the cytoplasm. By 16 h, these became more prominent and increased in number, and by 18-22 h they coalesced to enclose areas of the cytoplasm. These changes in the actin profile were confirmed by electron microscopy, where small actin bundles appeared in association with vesicles, increased in size, number and thickness, enclosed areas of cytoplasm with numerous vesicles and were finally seen in association with crystalline arrays of virus near the periphery of the cells. The addition of microfilament inhibitors cytochalasins B and D, after the initial period of adsorption resulted in complete inhibition of changes in the actin profile and of viral release, indicating that microfilament inhibitors prevented both polymerization of actin and movement of the virus within the cell. INTERPRETATION & CONCLUSION: In poliovirus infection, both intracellular movement and release of virus appear to be related to cytoskeletal changes, particularly involving actin microfilaments.

Adherent Escherichia coli in colorectal mucosal biopsies: a histological and ultrastructural evaluation.

BACKGROUND: Colorectal mucosal biopsies occasionally demonstrate the presence of bacteria adherent to the epithelium. This study evaluated the histological and ultrastructural correlates of such bacterial adherence. MATERIALS AND METHODS: Rectal mucosal biopsies from eight patients in whom histopathological examination of biopsies had earlier demonstrated adherent bacteria were examined by electron microscopy and by bacterial culture. Colorectal biopsies of 69 patients with adherent bacteria detected histologically were retrospectively evaluated for histological changes at sites proximal and distant to adherent bacteria. RESULTS: Escherichia coli of different serogroups were isolated from 7 of 8 rectal biopsies demonstrating bacterial adherence. All isolates showed diffuse or focal adherence to HEp-2 cell monolayers. Ultrastructural changes noted included microvillus damage, pedestal formation, actin web condensation, and protrusions of the apical cytoplasm of epithelial cells into the lumen towards the bacteria. Histological changes noted at light microscopy included reduction in epithelial cell height, focal epithelial cell degeneration, cryptitis and neutrophil infiltration at sites of bacterial adherence whereas these were usually absent at sites distant to adherent bacteria. Bacterial adherence was noted more often in biopsies from Crohn's disease patients than in patients without this diagnosis (P<0.001). CONCLUSION: Adherent Escherichia coli in colorectal biopsies were associated with focal epithelial damage and showed an association with Crohn's disease.

Risk of colorectal cancer in ulcerative colitis in India.

BACKGROUND: The risk for colorectal cancer (CRC) in ulcerative colitis (UC) in India is not known. METHOD: Retrospective cohort from a tertiary level hospital in South India. Analysis of archived records of all patients with UC who underwent colonoscopy and segmental biopsies over the last 25 years. Incidence densities and risk of developing high grade dysplasia or CRC was calculated and chi-squared test was performed for risk factors of interest. RESULTS: Complete records were available for 532 patients, 336 (63.2%) male. The mean (+/- SEM) duration of illness was 6.04 +/- 0.29 years. In total, 234 patients (44%) had pancolitis, 121 (22.7%) had left-sided colitis and 177 (33.3%) had proctitis or proctosigmoiditis. Overall, five (0.94%) patients developed carcinoma and one (0.19%) patient had high grade dysplasia. The incidence density and risk of developing either CRC or high grade dysplasia was zero in the first 10 years of disease. In those with disease duration of 10-20 years, incidence density was 2.34 per 1000 person years' duration (PYD) for all patients with colitis and 4.5 per 1000 PYD for patients with pancolitis alone. This corresponded to risks of 2.3% and 4.4%, respectively. For those with disease duration longer than 20 years, incidence density was 2.73 per 1000 PYD for all patients and 4.9 per 1000 PYD for patients with pancolitis. This corresponded to risks of 5.8% and 10.2%, respectively. Duration of disease beyond 10 years and extent of colitis were the only risk factors significantly associated with CRC. CONCLUSIONS: The risk of developing CRC is Indian patients with UC is lower than that reported from the West. Strategies for cancer surveillance in Indian patients with UC need to be tailored accordingly.

Vascular changes in duodenal mucosa in shigellosis and cholera.

Vascular endothelial cells are highly specialized cells with numerous sensory and modulator functions. Our previous studies show extensive microvascular changes in rectal mucosal vasculature of patients with acute infective diarrhea (Mathan and Mathan 1985a, Gut 26:710-717). We looked for changes in the duodenal mucosal vasculature in two naturally occurring diarrheal infections: shigellosis and cholera. Duodenal mucosal biopsies from 14 patients with shigellosis, 12 patients with cholera, and 10 healthy volunteers were examined under the electron microscope. There were extensive microvascular changes in the duodenum in shigellosis and cholera. Congestion and dilatation of capillaries and venules, stagnation of blood, thinning of the endothelial lining, and platelet clumping were commonly seen in both conditions. Endothelial damage was also common to both conditions but was mild to moderate in cholera and severe in shigellosis with frank hemorrhage, frequent formation of stress fibers, widening of intercellular spaces, cytoplasmic blebbing, cell fragmentation, and intravascular thrombosis. Erythrocyte aggregates, platelet aggregates, and leucocyte plugging lead to capillary obstruction. The arterioles were severely constricted. These changes in the endothelial lining of the microvasculature could contribute to the pathogenesis of the disease resulting in peripheral vascular insufficiency, inadequate oxygen delivery to intestine, and organ dysfunction. The factors influencing these changes, their implications, and possible therapeutic interventions are discussed.

Persistence of mucosal mast cells and eosinophils in Shigella-infected children.

Cells of the innate immune system and their mediators were studied at the single-cell level in the rectums of pediatric and adult patients with Shigella infection to better understand why children are at higher risk for severe infection. Adult patients had increased infiltration of mucosal mast cells (MMC) at the acute stage (3 to 5 days after the onset of diarrhea) and eosinophils in early convalescence (14 to 16 days after onset). Increased expression of stem cell factor and prostaglandin H synthase-1 (PGHS-1) was associated with increased tryptase-K(i)67-double-positive MMC in the acute stage and increased apoptosis of MMC, which led to a rapid decline in early convalescence. The eosinophils demonstrated increased expression of major basic protein (MBP), eotaxin, and CCR3, as well as increased necrotic death. The neutrophils showed enhanced alpha-defensin and lactoferrin expression in the acute phase. In contrast to adults, the pediatric patients demonstrated delayed accumulation of mast cells and eosinophils, while alpha-defensin expression persisted during convalescence. In contrast, neutrophil counts and lactoferrin expression were reduced in children compared to adults. The results suggest that children with shigellosis have a persistent activation of the innate immune response in the convalescent phase, indicating delayed elimination of Shigella antigens compared to adults.

Increased levels of inflammatory mediators in children and adults infected with Vibrio cholerae O1 and O139.

Investigations were carried out to study the production of factors associated with the innate immune response in the systemic and mucosal compartments in adults and children infected with Vibrio cholerae O1 and V. cholerae O139. The levels of nonspecific mediators of the innate defense system, i.e., prostaglandin E(2) (PGE(2)), leukotriene B(4) (LTB(4)), and lactoferrin (Lf), as well as myeloperoxidase (MPO), were elevated at the acute stage of the disease in stools obtained from both O1- and O139-infected adults and children. In the systemic compartment, the levels of Lf were increased after onset of disease, which in children remained elevated up to convalescence compared to the healthy controls. Increased concentrations of C-reactive protein were seen in the sera of adult cholera patients at the acute stage of infection. Elevated levels of the nitric oxide (NO*) metabolites (nitrite and nitrate [NO(2)(-) and NO(3)(-)]) were detected in plasma but not in urine. The activity of the scavenger of reactive oxygen species, superoxide dismutase, was higher in the plasma of adults immediately after the onset of disease, suggesting that an active scavenging of reactive oxygen species was taking place. The concentration of 8-iso-prostaglandin F(2 alpha) remained unchanged in the systemic and mucosal compartments in the study subjects. After the recovery of patients from cholera, the concentration of the majority of the metabolites decreased to baseline levels by day 30 after the onset of infection. Immunohistochemical staining showed increased tissue expression of MPO, Lf, and inducible nitric oxide synthase at the acute stage in the duodenal biopsies of adults and rectal biopsies obtained from children with cholera. Very little difference was seen in the levels of the different inflammatory mediators in patients infected with V. cholerae O1 or the encapsulated V. cholerae O139. In summary, these results suggest that elevated concentrations of Lf, MPO, PGE(2), LTB(4), and NO*, as well as other metabolites, during the acute stage of the disease indicate that the innate defense system, as well as the inflammatory process, is activated in both adults and pediatric patients infected with V. cholerae O1 and O139.

Adaptive and inflammatory immune responses in patients infected with strains of Vibrio parahaemolyticus.

In patients with diarrhea caused by Vibrio parahaemolyticus, antibody-secreting cell responses to thermostable direct hemolysin (TDH), lipopolysaccharide (LPS), and whole-cell bacteria were seen. TDH- and LPS-specific responses were seen in serum samples, and immunoglobulin A antibody responses were observed in stool. Levels of C-reactive protein and nitric oxide metabolites increased in the systemic circulation at the onset of illness. Tumor necrosis factor-alpha and lactoferrin levels were high during the acute stage in mucosal secretions and in plasma, whereas interleukin-1beta levels were high only in mucosal secretions. Duodenal and rectal biopsy specimens obtained at the onset of illness showed an acute inflammatory response. The lamina propria showed edema, congestion of blood vessels, and hemorrhage, with an increase in levels of polymorphonuclear neutrophils and macrophages. Strains belonging to different serotypes exhibited varying resistance to killing by serum; the O8:K21 strain was most sensitive. Infection with V. parahaemolyticus results in B cell responses and an acute inflammatory response that is self-limiting.