Hiroaki Itou

Systemic and local evidence of increased Fas-mediated apoptosis in ulcerative colitis

Abstract. Background and aims: Recent studies suggest that Fas-mediated apoptosis is involved in the pathogenesis of inflammatory bowel disease (IBD). This study was conducted to clarify whether soluble forms of Fas (sFas) and Fas ligand (sFasL) are concerned with inflammation in IBD. Methods and patients: Concentration of serum sFas and sFasL was measured by enzyme-linked immunosorbent assay in 10 patients with ulcerative colitis (UC), 10 with Crohns disease (CD) in both active and remission stages, and 20 controls. Expression of Fas and sFas in colonic mucosa was examined by western blot. Distribution of Fas and FasL in colonic mucosa was examined by immunohistochemistry in 20 UC, 20 CD, and 10 non-IBD colitis patients and in 10 controls. Apoptotic cells were examined by TUNEL. Results: Concentration of systemic sFas was significantly lower in active UC than controls. The number of FasL-containing cells was significantly higher in active UC than in remission UC, non-IBD colitis, and controls. Apoptotic cells were increased in active UC. Conclusions: Our results demonstrate that systemic and local Fas-mediated apoptosis is promoted in UC, which might be involved in the pathogenesis in UC.

Infliximab as a treatment for systemic amyloidosis associated with Crohn's disease

Systemic AA amyloidosis is a serious and potentially fatal complication of Crohn’s disease.1 This serious complication is associated with approximately 0.9–2.5% of patients with Crohn’s disease but renal failure due to renal amyloidosis is one of the most common causes of death for patients with Crohn’s disease.2 Although drugs such as azathioprine, colchicines, and dimethylsulphoxide are suggested to delay the progression of renal amyloidosis, the efficacy of these drugs on renal amyloidosis has not been fully elucidated.2 Infliximab is a chimeric anti-tumour necrosis factor α (TNF-α) monoclonal antibody. This drug has been proven …

Novel Evidence Suggesting Clostridium difficile Is Present in Human Gut Microbiota More Frequently than Previously Suspected

Prevalence rate of Clostridium difficile in healthy human adults is believed to be very low. Our RT‐PCR system using glass powder, which can eliminate PCR inhibitors, detected C. difficile toxin B mRNA in 16 of 30 fecal samples (53.3%) from healthy human adults. In contrast, we failed to detect toxin B in the same fecal samples by PCR using DNA templates extracted with phenol‐chloroform. Our results suggest that PCR inhibitors in feces carried through phenol‐chloroform extraction procedure might suppress the sensitivity of PCR and that C. difficile is actually present in human gut microbiota more frequently than previously suspected.

No evidence of persistent mumps virus infection in inflammatory bowel disease

BACKGROUND AND AIM There is controversy regarding whether paramyxovirus infection is causally associated with inflammatory bowel disease (IBD). The latest cohort study claimed that atypical measles and mumps infections in childhood may be risk factors for later IBD. This study was conducted to clarify the validity of a causal link between persistent mumps virus infection and IBD. SUBJECTS AND METHODS (1) Amplification of the mumps virus genome was performed in both intestinal specimens (ulcerative colitis 15, Crohns disease 15, control 10) and peripheral blood lymphocytes (PBL) (ulcerative colitis seven, Crohns disease six, control three) by reverse transcription-polymerase chain reaction (RT-PCR) followed by Southern hybridisation using primers specific to the viral genome encoding phosphoprotein or haemagglutinin-neuraminidase. (2) Titre of serum antimumps IgG was measured in 16 patients with ulcerative colitis, in 16 patients with Crohns disease, and in 16 normal controls using an enzyme linked immunosorbent assay. RESULTS (1) The mumps virus genome was not detected by RT-PCR in intestinal specimens or PBL in any case. (2) Antimumps IgG titre was positive in 7/16 ulcerative colitis, 10/16 Crohns disease, and 11/16 control specimens. The mean (SEM) titre of antimumps IgG was 12.281 (7.831) in ulcerative colitis, 7.675 (1.608) in Crohns disease, and 8.637 (1.969) in controls, with no significant difference between the three groups. CONCLUSION We could not find any evidence to support a causal link between persistent mumps virus infection and IBD.

Altered expression of epimorphin in ulcerative colitis.

Background and Aim: Epimorphin is suggested to play a key role in the morphogenesis of epithelial cells. We focused on epimorphin and attempted to clarify the relationship between expression of this molecule and inflammatory bowel disease (IBD).

Efficacy of ecabet sodium enema on steroid resistant or steroid dependent ulcerative colitis

Ecabet sodium (ES), a 12-sulfodehydroabietic acid monosodium salt derived from an ingredient in pine resin, has been shown to topically enhance several gastric mucosal defensive factors.1 ES is widely used for the treatment of gastritis and gastric ulcer in Japan. A recent open label study demonstrated that ES also has therapeutic effects in active ulcerative colitis (UC).2 In that study, the authors demonstrated that six of seven UC patients with proctosigmoiditis achieved complete remission by treatment with ES enema. However, successive clinical trials of the efficacy of ES enema in UC patients have not been reported. Here we demonstrate the efficacy of ES enema in steroid resistant or steroid dependent UC patients. …

Sucralfate prevents the delay of wound repair in intestinal epithelial cells by hydrogen peroxide through NF-κB pathway

BackgroundRecent studies have shown that sucralfate (SF) has therapeutic effects on colonic inflammation in ulcerative colitis. The aim of this study was to clarify the function of SF for wound repair in intestinal epithelial cells (IEC).Methods(1) Activation of signal proteins [ERK1/2 mitogen-activated protein kinase (MAPK), IκB-α] in IEC-6 cells after stimulation with 10−4 M potassium sucrose octasulfate (SOS), which is the functional element of SF, was assessed by Western blot. (2) Induction of transforming growth factor (TGF)-β1, TGF-α, EGF, and cyclooxygenase-2 (COX-2) mRNA after stimulation of IEC-6 cells with SOS was assessed by reverse transcriptase-polymerase chain reaction. (3) IEC-6 cells were wounded and cultured for 24 h with various concentrations of SOS in the absence or presence of 20 μM H2O2. Epithelial migration or proliferation was assessed by counting migrating cells or bromodeoxyuridine (BrdU)-positive cells across the wound border.Results(1) SOS activated IκB-α, but it did not activate ERK1/2 MAPK. (2) SOS enhanced the expression of COX-2 mRNA, but it did not change the mRNA expression of other growth factors. (3) SOS did not enhance wound repair in IEC-6 cells, but it decreased the number of dead cells (maximum, 74%) (P < 0.01) in a dose-dependent manner and prevented the diminishment of epithelial migration (maximum, 61%) (P < 0.01) and proliferation (maximum, 37%) (P < 0.05) induced by H2O2. These functions of SOS were suppressed by the NF-κB and COX-2 inhibitors.ConclusionsSOS prevented the delay of wound repair in IEC-6 cells induced by H2O2, probably through induction of COX-2 and an anti-apoptotic mechanism. These effects of SOS might be given through the activation of the NF-κB pathway.

A case of pseudoobstruction of the intestine associated with scleroderma dramatically responding to antibiotics.

To the Editor: Scleroderma is a systemic disease of unknown etiology characterized by excessive deposition of collagen and other connective tissue components in the skin and multiple organs.1,2 Scleroderma is commonly complicated by gastrointestinal involvement, which is characterized by dilatation of the gastrointestinal tract and reduction of peristalsis, sometimes resulting in pseudoobstruction of the intestine.3 Because pseudoobstruction is intractable and relapses are frequent despite careful treatment, it is difficult to manage patients with pseudoobstruction. Malnutrition due to pseudoobstruction has been reported to be the primary causes of death in scleroderma.3 Thus, it is very important to manage the pseudoobstruction and rescue such patients. Here, we describe a patient with intractable pseudoobstruction of the intestine associated with scleroderma, who significantly responded to treatment with antibiotics. In April 2001, a 65-year-old man visited our hospital with swelling of the skin of the fingers and breast. He was diagnosed as having scleroderma on the basis of findings of macroscopic and microscopic sclerosis cutanea and Raynaud’s phenomenon. In September 2004, he became conscious of abdominal fullness, vomiting, and inability to defecate. He visited another hospital, which suggested that he suffered from pseudoobstruction of the intestine associated with scleroderma. He underwent decompression with a long intestinal tube and total parenteral nutrition with administration of metoclopramide. His clinical symptoms of the patient soon improved, but he experienced frequent relapses of pseudoobstruction. Thus, he was referred to our hospital on 4 December 2004. The physical examination on admission revealed abdominal distension, a decrease in bowel sounds, and stiffness and swelling of the skin of the hand and foot. Hematologic and serum chemistry showed hypoalbuminemia (3.0g/dl) and anemia (RBC, 245 × 104/μl, Hb 7.0 g/dl). Antinuclear antibody was strongly positive (1 : 320), with a speckled pattern. Antiscleroderma 70 antibody and anticentromere antibody were also positive (1 : 22.7 and 1 : 35.1, respectively). Fecal culture showed normal gut flora and no pathogenic bacteria. The plain film of the abdomen on admission demonstrated significantly dilated loops of small intestine with air–fluid levels (Fig. 1A). Abdominal ultrasonography also showed dilatation of loops of the small bowel. Endoscopic examination of the upper gastrointestinal tract and the colon and terminal ileum showed no obstructive or inflammatory lesions. The small intestine series demonstrated significant dilatation and hypomotility of the small intestine, but no mechanical obstruction was found (Fig. 1B,C). Based on these findings, we confirmed the diagnosis of pseudoobstruction of the intestine associated with scleroderma. Therefore, the patient was treated with dimeticon and a Chinese herbal drug (dai-kenchu-to; Tsumura, Tokyo, Japan), but the abdominal symptoms and X-ray findings did not improve. Taking into account the association of bacterial overgrowth with his symptoms, we added a treatment with kanamycin and lactulose. In response, the abdominal symptoms of the patient soon improved. However, abdominal symptoms recurred after 2 weeks. We changed antibiotics from kanamycin to metronidazole on the 28th hospital day because Lock et al.4 recommended treatment with metronidazole for pseudoobstruction associated with scleroderma. After the treatment with metronidazole (750 mg/day), the abdominal symptoms of the patient dramatically improved in 3 days, and the X-ray findings also significantly improved (Fig. 1D). He started oral ingestion of food on the 47th hospital day, but abdominal symptoms did not recur. He was discharged from our hospital on the 65th hospital day. Pseudoobstruction in the patient recurred 6 months later, when levofloxacin (300 mg/day) significantly improved his abdominal symptoms. This case suggests that bacterial overgrowth in the intestine might be closely involved with abdominal symptoms on the part of patients with intractable pseudoobstruction associated with scleroderma. For such patients, treatment with specific antibiotics such as metronidazole or new quinolones might improve their abdominal symptoms even when pathogenic bacteria cannot be identified in the patient’s feces.