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Background: Helicobacter pylori (HP) infection induces methylation silencing of specific genes in gastric epithelium. Various stimuli activate the nonselective cation channel TRPV4, which is expressed in gastric epithelium where it detects mechanical stimuli and promotes ATP release. As CpG islands in TRPV4 are methylated in HP-infected gastric epithelium, we evaluated HP infection-dependent changes in TRPV4 expression in gastric epithelium., Materials and Methods: Human gastric biopsy samples, a human gastric cancer cell line (AGS), and a normal gastric epithelial cell line (GES-1) were used to detect TRPV4 mRNA and protein expression by RT-PCR and Western blotting, respectively. Ca 2+ imaging was used to evaluate TRPV4 ion channel activity. TRPV4 methylation status was assessed by methylation-specific PCR (MSP). ATP release was measured by a luciferin-luciferase assay., Results: TRPV4 mRNA and protein were detected in human gastric biopsy samples and in GES-1 cells. MSP and demethylation assays showed TRPV4 methylation silencing in AGS cells. HP coculture directly induced methylation silencing of TRPV4 in GES-1 cells. In human samples, HP infection was associated with TRPV4 methylation silencing that recovered after HP eradication in a time-dependent manner., Conclusion: HP infection-dependent DNA methylation suppressed TRPV4 expression in human gastric epithelia, suggesting that TRPV4 methylation may be involved in HP-associated dyspepsia., (© 2016 The Authors. Helicobacter Published by John Wiley & Sons Ltd.)

Background:  High-molecular-weight cell-associated proteins (HM-CAP) assay is the most popular serological immunoassay worldwide and has been developed from US isolates as the antigens. The accuracy is reduced when the sera are from adults and children in East Asia including Japan. To overcome the reduced accuracy, an enzyme immunoassay using Japanese strain-derived HM-CAP (JHM-CAP) was developed, in which the antigens were prepared by exactly the same procedure as HM-CAP. The performance of JHM-CAP was better than that of HM-CAP in Japanese adults as well as in children. The higher sensitivity was because of the presence of 100-kDa protein that was absent in the preparation of HM-CAP antigen., Materials and Methods:  Immunoblot analysis and peptide mass fingerprinting methods were used to identify the distinctive 100-kDa protein present in JHM-CAP antigens. The peptide sequence and identification were analyzed by Mascot Search on the database of Helicobacter pylori. The identified protein was confirmed by immunoblot with a specific antibody and inhibition assay by the sera., Results: The distinctive 100-kDa protein was a fragment of CagA derived from Japanese clinical isolates, and the sera of Japanese patients had strongly reacted to the protein, probably to the exposed epitope on the fragmented CagA. The fragmentation of CagA had occurred in the process of antigen preparation in Japanese isolates, not in US isolates even under the same preparation., Conclusion:  The distinctive 100-kDa protein was a fragment of CagA protein of H. pylori derived from Japanese clinical isolates, and Japanese patients including children are likely to react strongly to the exposed epitopes on fragmented CagA., (© 2012 Blackwell Publishing Ltd.)

Problem: It is well known that the prevalence of several autoantibodies is higher in patients with recurrent miscarriages than in normal women. However, links between individual autoantibodies are unclear. The present study focuses on the possible association between beta 2-glycoprotein I (beta 2-GPI)-dependent anticardiolipin antibody (aCL), lupus anticoagulant (LA), and antinuclear antibody (ANA) in patients with recurrent miscarriages., Method of Study: Three hundred and one patients, with a history of two or more unexplained miscarriages, were studied. The titers of beta 2-GPI-dependent aCL and LA were then compared between single-antibody-positive and three-antibody-positive groups., Results: The prevalences of beta 2-GPI-dependent aCL, LA, and ANA were 3.3, 10.0, and 25.2%, respectively. Four of the 301 patients had all three antibodies. The LA titers in patients with positive values for three antibodies was significantly higher than in cases with only LA., Conclusion: beta 2-GPI-dependent aCL, LA, and ANA define three distinct, but partly related populations in patients with recurrent miscarriage. We should test at least two kinds of autoantibodies in recurrent aborters, because it has been found that, e.g., beta 2-GPI-dependent aCL and LA are predictors for miscarriages.

Problem: The purpose of this study was to determine whether the serum macrophage-colony stimulating factor (M-CSF) level is associated with early pregnancy loss in unexplained recurrent spontaneous abortion (RSA) patients., Method: We therefore compared preconceptional serum M-CSF levels between unexplained RSA patients and controls. The former comprised 44 bed-rest therapy patients and 43 intradermal immunization (IDI) patients receiving paternal lymphocyte therapy, who had experienced two and three or more consecutive first-trimester pregnancy losses, respectively. The controls were 46 healthy non-pregnant women. We also prospectively studied the association between M-CSF levels during pregnancy and adverse pregnancy outcomes. Sera from a total of 31 pregnant women, including 16 of the bed-rest therapy group and 15 of the IDI therapy group, were collected at the 4th, 6th, and 8th gestational weeks and were measured for M-CSF levels, using the enzyme-linked immunoadsorbent assay (ELISA) method established by Hanamura et al., Results: Serum M-CSF levels were significantly lower in the non-pregnant RSA patients (460.0 +/- 185.6 U/ml; mean +/- SD) than in the control group (726.5 +/- 134.0 U/ml) and also were lower at the 8th, but not the 4th or 6th gestational week in those patients of both the bed-rest and IDI therapy groups whose outcome was pregnancy failure., Conclusion: Thus a low level of serum M-CSF was found to be associated with unexplained recurrent pregnancy loss in both the preconceptional and conceptional phases. These results raise the possibility that M-CSF may play an important role in the maintenance of pregnancy and that it can be used as a parameter for determining individuals at risk.