Molecular cloning reveals nearly half of patients with Crohn's disease have an antibody to peroxiredoxin 6-like protein.

BACKGROUND AND AIM: Crohn's disease (CD) is a chronic inflammatory bowel disease (IBD) of unknown etiology. We aimed to identify the etiological agent of CD using a molecular cloning strategy that was particularly focused on identifying agents causing immune abnormalities and infectious agents. METHODS: We constructed a cDNA library derived from the inflamed intestinal tissue of a CD patient, and screened 1.5 million clones in this library with the serum from another typical CD patient. The expressed cDNA clones that positively reacted with the serum were then expressed as fusion proteins with glutathione S-transferase, and western blotting was performed using the sera of 22 CD, 13 ulcerative colitis (UC), and 16 non-IBD patients. RESULTS: We identified nine positive clones that did not contain any viral or bacterial genomic DNA. Of these, we selected one clone (clone 50) with which the typical CD patient's serum most strongly reacted. Clone 50 is highly homologous to the antioxidant protein peroxiredoxin 6. In western blotting, the sera of 47.6% CD patients (small intestine type 80%, large and small intestine type 43%, large intestine type 0%) showed strong reactivity to clone 50, none of the UC patients were reactive to clone 50, and 18.8% of non-IBD patients were very weakly reactive to it. We also found that the expression of peroxiredoxin 6 was significantly increased in inflamed intestinal epithelia of CD. CONCLUSION: The present study first showed that some CD patients have an antibody against peroxiredoxin 6-like protein, which may be involved in the pathogenesis of CD.

Cost-effectiveness analysis of a universal rotavirus immunization program in Japan.

In anticipation of the imminent licensure of rotavirus vaccine, we evaluated the cost-effectiveness of rotavirus vaccine in Japan by taking into account the considerable variations in the incidence of rotavirus-associated hospitalizations previously reported in the literature. We assumed that the variation was due to local differences in healthcare utilization practices rather than a true difference in the incidence of severe rotavirus gastroenteritis. Thus, a Markov model was constructed such that the sum of rotavirus-associated hospitalizations and outpatient visits was set a constant value of 129 cases per 1,000 child-years. We calculated the direct medical cost, the indirect cost, and the quality-adjusted life year (QALY) loss in children aged less than 5 years. For the base case scenario, the incremental cost-effectiveness ratio (ICER) per QALY gained was 9.8 million Japanese yen from the healthcare perspective, but it was 900,000 Japanese yen from the societal perspective, making the program of universal immunization against rotavirus highly cost-effective. Furthermore, the universal immunization program was found to be cost-effective from the societal perspective for any of the previously reported incidence rates of rotavirus-associated hospitalization. Thus, the introduction of the rotavirus vaccine into the childhood immunization schedule and its co-administration with other childhood vaccines will be a cost-effective public health intervention in Japan.

The bovine lactophorin C-terminal fragment and PAS6/7 were both potent in the inhibition of human rotavirus replication in cultured epithelial cells and the prevention of experimental gastroenteritis.

Rotaviruses are the leading cause of severe dehydrating diarrhea in children worldwide. We have found that high-M(r) glycoprotein fraction (F1) from cow's milk whey has potent inhibitory activity against human rotavirus (HRV) in cell culture. The present study was undertaken to identify and characterize the components responsible for this inhibitory activity. F1 was initially heated at 95 degrees C for 30 min, rendering milk antibodies inert, subjected to ammonium sulfate fractionation, and then resolved by two-dimensional polyacrylamide gel electrophoresis. After electroelution, we found that a heat-stable milk protein lactophorin C-terminal fragment (LP16) and bovine milk fat globule membrane protein PAS6/7 strongly inhibited the replication of HRV MO strains in MA104 cells. Furthermore, we found that prophylactic oral administration of F1 once before inoculation of the HRV MO strain obviously prevented the development of diarrhea in vivo. These non-immunoglobulin components are a promising candidate for a prophylactic food additive against HRV infection.

Healthcare-associated viral gastroenteritis among children in a large pediatric hospital, United Kingdom.

Viruses are the major pathogens of community-acquired (CA) acute gastroenteritis (AGE) in children, but their role in healthcare-associated (HA) AGE is poorly understood. Children with AGE hospitalized at Alder Hey Children's Hospital, Liverpool, UK, were enrolled over a 2-year period. AGE was classified as HA if diarrhea developed > or =48 hours after admission. Rotavirus, norovirus, adenovirus 40/41, astrovirus, and sapovirus were detected by PCR. A total of 225 children with HA-AGE and 351 with CA-AGE were enrolled in the study. HA viral gastroenteritis constituted one fifth of the diarrheal diseases among hospitalized children and commonly occurred in critical care areas. We detected > or =1 virus in 120 (53%) of HA-AGE cases; rotavirus (31%), norovirus (16%), and adenovirus 40/41 (15%) were the predominant viruses identified. Molecular evidence indicated rotaviruses and noroviruses were frequently introduced into the hospital from the community. Rotavirus vaccines could substantially reduce the incidence of HA-AGE in children.

Incidence of rotavirus and all-cause diarrhea in northeast Brazil following the introduction of a national vaccination program.

BACKGROUND & AIMS: Rotavirus vaccines were introduced in Brazil in 2006; we evaluated their effects in the state of Sergipe, Brazil. METHODS: We performed a cross-sectional survey of children with diarrhea attending emergency services in Aracaju, Brazil, between October 2006 and April 2008 and a cluster sampling survey to assess vaccination coverage. Vaccine efficacy was assessed using the screening method. Diarrhea consultation and hospitalization data (2003-2007) were obtained from state and national surveillance systems. RESULTS: Rotavirus was detected in 59 of 534 stool samples (11%) from children attending emergency services. The number of rotavirus-positive samples decreased from 18 of 74 (24%) in 2006 to 31 of 321 (9.5%) in 2007 and 10 of 136 (7.4%) in 2008 (P < .01). Diarrhea severity was greater in children with rotavirus (P < .01) but decreased over time (P < .001). Of the rotaviruses detected, 56 of 59 (95%) were P[4]G2 genotype, 1 was P[4]G-non-typeable (NT), 1 was P[NT]G2, and 1 was P[NT]GNT. Diarrhea consultations decreased from 3020 in 2004 to 604 in 2007; reductions were greatest among children under 5 years old. Diarrhea hospitalizations decreased from 2121 in 2003 to 1176 in 2007. Vaccine coverage was 90.3%. Vaccines were highly effective against the strain P[8]G1; efficacy against P[4]G2 genotype was 89% (95% confidence interval: 0.87-0.92) in Aracaju and 95% in Sergipe. CONCLUSIONS: Since vaccines were introduced in 2006, there has been an overall reduction in diarrhea consultations and hospitalizations in northeast Brazil, with the greatest reductions in young children. This might have resulted from vaccination and improved sanitation. Although a single rotavirus genotype (P[4]G2) was recovered, vaccine efficacy was high against this genotype.

Rotavirus double-stranded RNA induces apoptosis and diminishes wound repair in rat intestinal epithelial cells.

BACKGROUND: Recent studies have shown that toll-like receptor 3 (TLR3) recognizes double-stranded RNA (dsRNA). Rotaviruses, having a dsRNA genome, infect intestinal epithelial cells (IEC) and cause acute gastroenteritis in young children. The aim of the present study was to clarify the pathophysiological function of rotavirus dsRNA in IEC. METHODS: Expression of TLR3 mRNA or protein in IEC cell lines (IEC-6, HT-29, Caco-2) was assessed by reverse transcription polymerase chain reaction (RT-PCR), Western blot analysis or immunohistochemistry. Induction of cytokines (TNF-alpha, interferon-beta, interleukin-6) mRNA and activation of signal proteins (ERK1/2 MAPK and IkappaB-alpha) in IEC after stimulation with rotavirus dsRNA were assessed by RT-PCR or Western blot analysis. IEC-6 cells were wounded and cell migration into wound areas after stimulation with rotavirus dsRNA (1-25 microg/mL) was assessed. Induction of apoptosis after stimulation with rotavirus dsRNA was also assessed. RESULTS: Expression of TLR3 mRNA and TLR3 protein was detected in IEC. Expression of TLR3 mRNA in IEC-6 tended to be up-regulated by exposure to IFN-gamma. Induction of cytokine mRNA and activation of the signal proteins were detected after stimulation with rotavirus dsRNA. Apoptosis was induced and epithelial migration into the wound area was dose-dependently diminished (44.1-94.4%, P < 0.01) by exposure to rotavirus dsRNA. Diminishment of wound repair was suppressed by anti-TLR3 antibody or caspase inhibitor. CONCLUSION: Rotavirus dsRNA induces severe apoptosis and diminishes wound repair in IEC through TLR3, which might be involved in the pathogenesis of rotavirus-induced enteritis.