Tohru Fujii

Bifidobacterium breve enhances transforming growth factor beta1 signaling by regulating Smad7 expression in preterm infants.

Objectives: Transforming growth factor (TGF) &bgr;1 displays a broad spectrum of activities in mucosal regulation, including induction of oral tolerance, potent anti-inflammatory effects, mucosal IgA expression and effects on epithelial cell proliferation and differentiation. The present study examined the effect of probiotics on the immunologic system of preterm infants in relation to TGF-&bgr; signaling. Methods: Subjects comprised 19 preterm infants divided into 2 groups: receiving Bifidobacterium breve supplementation (B. breve group) and without supplementation (controls). Blood samples were collected from both groups on days 0, 14 and 28 after birth. Serum cytokine levels were measured using enzyme-linked immunosorbent assay, and expression levels of the TGF-&bgr; signaling molecule, Smad, were examined using semiquantitative reverse transcriptase-polymerase chain reaction. Results: Serum TGF-&bgr;1 level was elevated on day 14 and remained elevated on day 28 in the B. breve group. Level of messenger RNA expression was enhanced for Smad3 and reduced for Smad7 (antagonistic Smad) after B. breve administration relative to levels in controls on day 28. Conclusions: These results demonstrated that the administration of B. breve to preterm infants can up-regulate TGF-&bgr;1 signaling and may possibly be beneficial in attenuating inflammatory and allergic reactions in these infants.

Effects of highly purified eicosapentaenoic acid on erythrocyte fatty acid composition and leukocyte and colonic mucosa leukotriene B4 production in children with ulcerative colitis.

Background n-3 Polyunsaturated fatty acids (PUFAs) have been suggested as a treatment for ulcerative colitis (UC). However, the efficacy of n-3 PUFAs against UC has not been examined in children. Therefore, the authors investigated the effects of eicosapentaenoic acid (EPA) on fatty acid composition and leukotriene (LT) production in children with UC. Methods For 2 months the authors administered highly purified EPA ethyl ester (EPA-E) (1.8 g/d) to children with UC in remission. Colonic mucosal histology, fatty acid composition of erythrocyte membrane phospholipids, and LTB4 production by leukocytes and colonic mucosa were measured before and 2 months after the initiation of EPA-E treatment. Results No patients relapsed during the study period, and no significant differences were detected in laboratory findings obtained before and 2 months after the initiation of EPA-E ingestion. There were no significant differences in mucosal histologic scores before and 2 months after EPA-E treatment. The EPA levels in erythrocyte membranes 2 months after the initiation of EPA-E treatment were significantly higher than before treatment, but the other fatty acids showed no significant changes. LTB4 production by leukocytes and rectal mucosa after 2 months of EPA-E treatment was significantly lower than before treatment. Conclusion EPA-E treatment increased the levels of EPA in erythrocytes and decreased LTB4 levels produced by leukocytes and colonic mucosa. To assess the concomitant clinical changes, we should examine the long-term effects of EPA-E ingestion on the maintenance of remission in children with UC.

Increased mucosal expression of GATA-3 and STAT-4 in pediatric ulcerative colitis

Background:  Serum pro‐inflammatory cytokine levels are frequently elevated in the acute phase of pediatric inflammatory bowel disease (IBD). Because the role of pro‐inflammatory cytokine in the acute phase of pediatric IBD has not been well investigated, the serum levels of pro‐inflammatory cytokines and the expression of Th1 and Th2 signaling molecules in mucosa from the acute phase of pediatric IBD were examined.

Neonatal transient eosinophilic colitis causes lower gastrointestinal bleeding in early infancy.

Background: Lower gastrointestinal bleeding (LGB), particularly in newborns, is of serious concern and requires urgent investigation and hospital care. Whereas allergic proctocolitis caused by food protein is a significant cause of LGB in infants with eosinophilia, there are several cases of diseases with symptoms similar to those of allergic proctocolitis but without an apparent allergic reaction influence. Patients and Methods: We examined 2 neonates using rectosigmoidoscopy who showed eosinophilia and experienced fresh LGB soon after birth and before their first feedings. Serum eosinic cationic protein (ECP) and platelet activating factor (PAF) levels were also examined in the second case to confirm the involvement of eosinophils for its pathogenesis. Results: Both patients were in a clinically stable condition, and their abdomens were soft. The results of their blood analyses, abdominal radiographs, and stool cultures were normal, but they had gross eosinophilia: the eosinophil counts were 9014/mm3 (patient 1) and 1955/mm3 (patient 2). Rectosigmoidoscopy with colonic mucosal biopsy revealed nodular lymphoid hyperplasia with a pale mucosal surface and massive oozing with diffuse eosinophilic infiltration in the lamina propria. In patient 2 the serum ECP and PAF levels were elevated to 123 μg/L (normal, <14.7) and 13.1 μmol/L/min (normal, <6). A few days after intravenous hydration therapy, LGB was no longer detected, and the serum ECP and PAF levels returned to normal. Conclusions: Inasmuch as these infants had LGB similar to allergic proctocolitis without any allergic reactions, we suggest that infiltrated eosinophils in the colonic mucosa could be involved in the pathogenesis of LGB in early infancy.

Fecal α1-antitrypsin Concentrations as a Measure of Enteric Protein Loss After Modified Fontan Operations

Background Little is known about the enteric protein loss in patients after a modified Fontan operation before the appearance of overt symptoms or signs of protein-losing enteropathy (PLE). The authors examined the possibility of using fecal &agr;1-antitrypsin concentration measurements for the early detection of postoperative PLE and in longer term postoperative monitoring of these patients. Methods The authors compared fecal &agr;1-antitrypsin concentrations in stool samples from 12 children 12.0 to 43.7 months after modified Fontan operations with those of 12 age-matched control subjects and examined the relationship between the &agr;1-antitrypsin levels and time since operation. The authors also compared fecal &agr;1-antitrypsin concentrations of stools from the same patients obtained at two different time points after surgery with intervals between samples ranging from 14.7 to 19.8 months. Results No significant differences in serum total protein and albumin levels were observed between patients after the modified Fontan operation and control subjects. The fecal concentrations of &agr;1-antitrypsin in patients after the Fontan operation were significantly (P < 0.01) higher than those in control subjects. There was no significant correlation between fecal &agr;1-antitrypsin concentrations and time elapsed after the Fontan operation. The fecal &agr;1-antitrypsin concentration increased significantly (P < 0.01) over periods of 14.7 to 19.8 months after the first measurement. Conclusion The results show that enteric protein loss begins before the appearance of hypoproteinemia in patients after a modified Fontan operation, and that the measurement of fecal &agr;1-antitrypsin concentrations in random stool samples is useful as an early indicator. To watch for the development of PLE after Fontan operation, it may be important to perform longitudinal follow-up examinations of enteric protein loss by measuring fecal &agr;1-antitrypsin concentrations early in the postoperative period.

ω-3 fatty acids attenuate mucosal inflammation in premature rat pups.

BACKGROUND Necrotizing enterocolitis (NEC) is a devastating intestinal disease of premature infants. Although ω-3 fatty acids are known to have antiinflammatory effects, their effect against NEC remains unclear. METHODS Mother rats fed a soybean-based, docosahexaenoic acid (DHA)- or eicosapentaenoic acid (EPA)-enriched diet from days 7 to 20 of gestation were examined. On day 20, the rat pups were delivered by abdominal incision, their intestines were removed, and messenger RNA was extracted. A rat NEC model was used to confirm the effects of ω-3 fatty acids on the inflamed intestine (n = 20-28). The expression of inflammatory molecules was analyzed by real-time polymerase chain reaction (n = 11-14). RESULTS The concentrations of DHA and EPA in the intestine were significantly increased in the DHA and EPA groups (P < .01). The expression of the antiinflammatory prostaglandin E2 receptor EP3 was increased in the DHA (P < .05) and EPA groups (P < .01). In the NEC model, the reduced incidence of colitis was confirmed in the DHA and EPA groups. The expression of peroxisome proliferator-activated receptor γ was increased (P < .05), and the inhibitor of nuclear factor-κB α/β decreased in both the DHA (P < .01) and EPA groups (P < .05). CONCLUSION Our findings indicate that ω-3 fatty acids are beneficial for protecting the premature intestine from inflammation by regulating eicosanoid- and nuclear factor-κB-related metabolite expression.

Enhanced production of interferon-gamma as a possible cause of protein-losing enteropathy after modified Fontan operation.

Protein-losing enteropathy (PLE) is a serious complication after modified Fontan operations in patients with congenital heart diseases (1–5). High central venous pressure with consequent intestinal lymphangiectasia has been suggested as the principal cause of PLE, but other potential contributing factors, such as inflammatory processes, have also been considered (1,4,5). Corticosteroid treatment for patients with PLE after a Fontan procedure has been reported as an effective therapy (1,4,5). Although the precise mechanism of action of corticosteroids is unclear, their effectiveness suggests that autoimmune and inflammatory processes may be one of the pathogenic factors of PLE. The local immune and inflammatory responses have not been fully investigated after Fontan procedure, and, to our knowledge, there has been no study of cytokine levels or electron microscopic findings in the small intestinal mucosa of patients with PLE after a Fontan operation. We report a boy who developed PLE after a modified Fontan operation. A jejunal biopsy was performed in this child before and after successful corticosteroid treatment of his PLE. Histologic analysis and measurement of the cytokine levels in the small intestinal mucosa were performed.

Monitoring 6-thioguanine nucleotide concentrations in Japanese children and adolescents with inflammatory bowel disease.

Background and Aim:  6‐Mercaptopurine (6‐MP) and azathioprine (AZA) are widely used as maintenance therapy in children with inflammatory bowel disease (IBD). However, proper 6‐thioguanine nucleotide (6‐TGN) concentrations in Japanese children with IBD have not been reported.

Changes in the presence of urine Helicobacter pylori antibody in Japanese children in three different age groups

Background: The rates of acquisition and spontaneous eradication of Helicobacter pylori infection in children has yet to be established. To determine these rates in children living in an urban region of Japan, the levels of urine H. pylori antibodies in children of three different age groups were measured.

Pranlukast regulates tumour growth by attenuating IL-4 production in Kimura disease

The administration of pranlukast may regulate tumour growth and reduce allergic symptoms in patients with Kimura disease. Kimura disease is a chronic eosinophilic inflammatory disorder characterised by soft tissue tumours and enlarged lymph nodes in the head and cervical region, with increased peripheral eosinophil counts and elevated serum IgE levels. We describe two patients with Kimura disease who showed clinical improvement after pranlukast treatment. Case 1 was a 15-year-old boy suffering from a right cervical tumour, bronchial asthma, atopic dermatitis, and frequent urticaria. He was diagnosed as having Kimura disease (biopsy) at age 11 years. Tumour regression was observed following corticosteroid treatment and surgery, but the tumour never completely disappeared and re-appeared when treatment was stopped. Case 2 was a 12-year-old boy with a left cervical tumour. He had a past history of bronchial asthma, Henoch-Schönlein purpura, and frequent urticaria. He was diagnosed as having Kimura disease (biopsy) at age 10 years. Both patients were subsequently treated with pranlukast at a dosage of 450 mg/day. Reduced mass sizes, allergic symptoms, decreased eosinophil counts, and serum IgE levels were confirmed after 2 years of treatment in case 1 and 2 months in case 2. In contrast to corticosteroids, there were no side-effects observed during the treatment period. In addition, serum cytokine concentrations were evaluated using a commercially available ELISA kit (R&D Systems Inc., Minneapolis, MN) in case 2. Elevation of serum IL-4 (more than 300 times higher than normal), IL-5, and IL-6 levels before treatment and decreased serum IL-4 levels after treatments were confirmed (Table 1). Since these Th2 type cytokines are known to promote IgE production, eosinophil migration, and Th2 cell activation, it is understandable that patients with Kimura disease often exhibit high serum IgE levels and increased eosinophil counts. Pranlukast, a cysteinyl LT-1 receptor antagonist, is well known to have a clinical benefit in the treatment and management of allergic diseases. Several studies have suggested that pranlukast inhibits LPS-induced IL-6 and allergen-specific TNF-a and Th2 type cytokine production and the translocation of NF-jB, a ubiquitous transcription factor for genes that encode proinflammatory cytokines such as IL-1, IL-6, IL-8, and TNF-a [2, 3, 4]. Most of these reports are in-vitro studies and there are only a few papers showing successful reduction in serum cytokine levels in clinical trials with allergic patients. The present study demonstrated that the serum IL-4 level was markedly decreased to 50% of the initial value following the pranlukast treatment in Kimura disease patients. Interestingly there was almost no change in other Th2 cytokine levels after pranlukast treatment. A recent study has suggested that intracrine cysteinyl LT receptor-mediated signalling induced by LTC4 acts as an intracrine mediator of eosinophil IL-4 secretion [1]. Our findings suggest that pranlukast is beneficial in the clinical management of Kimura disease, possibly by blocking this intracrine cysteinyl LT receptor signalling and attenuating IL-4 production leading to tumour regulation, decreased eosinophil counts, and decreased levels of serum IgE and may reduce clinical allergic symptoms in these patients. Y. Ohtsuka (&) Æ T. Shimizu Æ T. Fujii Æ M. Saito Æ S. Nagata Y. Yamashiro Department of Paediatrics, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, 113-8421 Tokyo, Japan E-mail: yohtsuka@med.juntendo.ac.jp Tel.: +81-3-58021075 Fax: +81-3-58000216