Role of the K(Ca)3.1 K+ channel in auricular lymph node CD4+ T-lymphocyte function of the delayed-type hypersensitivity model.

BACKGROUND AND PURPOSE: The intermediate-conductance Ca(2+)-activated K(+) channel (K(Ca)3.1) modulates the Ca(2+) response through the control of the membrane potential in the immune system. We investigated the role of K(Ca)3.1 on the pathogenesis of delayed-type hypersensitivity (DTH) in auricular lymph node (ALN) CD4(+) T-lymphocytes of oxazolone (Ox)-induced DTH model mice. EXPERIMENTAL APPROACH: The expression patterns of K(Ca)3.1 and its possible transcriptional regulators were compared among ALN T-lymphocytes of three groups [non-sensitized (Ox-/-), Ox-sensitized, but non-challenged (Ox+/-) and Ox-sensitized and -challenged (Ox+/+)] using real-time polymerase chain reaction, Western blotting and flow cytometry. KCa 3.1 activity was measured by whole-cell patch clamp and the voltage-sensitive dye imaging. The effects of K(Ca)3.1 blockade were examined by the administration of selective K(Ca)3.1 blockers. KEY RESULTS: Significant up-regulation of K(Ca)3.1a was observed in CD4(+) T-lymphocytes of Ox+/- and Ox+/+, without any evident changes in the expression of the dominant-negative form, K(Ca)3.1b. Negatively correlated with this, the repressor element-1 silencing transcription factor (REST) was significantly down-regulated. Pharmacological blockade of K(Ca)3.1 resulted in an accumulation of the CD4(+) T-lymphocytes of Ox+/+ at the G0/G1 phase of the cell cycle, and also significantly recovered not only the pathogenesis of DTH, but also the changes in the K(Ca)3.1 expression and activity in the CD4(+) T-lymphocytes of Ox+/- and Ox+/+. CONCLUSIONS AND IMPLICATIONS: The up-regulation of K(Ca)3.1a in conjunction with the down-regulation of REST may be involved in CD4(+) T-lymphocyte proliferation in the ALNs of DTH model mice; and K(Ca)3.1 may be an important target for therapeutic intervention in allergy diseases such as DTH.

[Quantitative analysis of hepatitis B surface antigen as a clinical marker].

There are 1.5 million hepatitis B virus (HBV) infected patients in Japan. Anti-viral therapy is important for chronic hepatitis B patients to prevent hepatocellular carcinoma. Recently, HBs antigen (HBsAg) quantification has been reported to be useful for not only HBV screening but also for monitoring of anti-viral treatment. In this paper, we evaluated the clinical utility of quantitative assay of HBsAg by HISCL HBsAg kit. Although there can be a significant difference in age, HBeAg positive/negative and viral genotype, there is not in the disease stage. Moreover, the weak correlation was confirmed between HBsAg and HBV-DNA levels with or without anti-virus treatment. In the clinical practice, as HBV-DNA becomes undetectable immediately by anti-viral therapy such as entecavir, it may be difficult to evaluate the efficacy. The monitoring of the HBsAg concentration in addition to HBV-DNA would be useful for the evaluation. Hence, the clinical role of HBsAg concentration could spread widely in Japan.

Frequent silencing of a putative tumor suppressor gene melatonin receptor 1 A (MTNR1A) in oral squamous-cell carcinoma.

Array-based comparative genomic hybridization (array-CGH) has good potential for the high-throughput identification of genetic aberrations in cell genomes. In the course of a program to screen a panel of 21 oral squamous-cell carcinoma (OSCC) cell lines for genome-wide copy-number aberrations by array-CGH using our in-house bacterial artificial chromosome arrays, we identified a frequent homozygous deletion at 4q35 loci with approximately 1 Mb in extent. Among the seven genes located within this region, the expression of the melatonin receptor 1 A (MTNR1A) messenger RNA (mRNA) was not detected or decreased in 35 out of the 39 (89%) OSCC cell lines, but was detected in immortalized normal oral epithelial cell line, and was restored in gene-silenced OSCC cells without its homozygous loss after treatment with 5-aza-2'-deoxycytidine. The hypermethylation of the CpG (cytosine and guanine separated by phosphate) island in the promoter region of MTNR1A was inversely correlated with its expression in OSCC lines without a homozygous deletion. Methylation of this CpG island was also observed in primary OSCC tissues. In an immunohistochemical analysis of 50 primary OSCC tumors, the absence of immunoreactive MTNR1A was significantly associated with tumor size and a shorter overall survival in patients with OSCC tumors, and seems to be an independent prognosticator in a multivariate analysis. Exogenous restoration of MTNR1A expression inhibited the growth of OSCC cells lacking its expression. Together with the known tumor-suppressive function of melatonin and MTNR1A in various tumors, our results indicate MTNR1A to be the most likely target for epigenetic silencing at 4q35 and to play a pivotal role during oral carcinogenesis.

Increased inhibition and decreased facilitation effect during a lexical decision task in children.

Widespread theories propose that implicit memory is established in the early stages of development, while explicit memory continues to develop until later stages. However, recent studies have argued that implicit memory changes developmentally. In order to elucidate the differences of implicit memory and semantic memory structures between children and adults, a single-word presentation method was developed using lexical decision tasks in which repeated real words, related real words, unrelated real words and pseudowords were presented in a list. Semantic priming and repetition priming using a single-word presentation method were employed in 25 children and 18 adults. Reaction time (RT) and correct rate for real words following pseudowords served as the base. Inhibition effect was defined as an increase in RT between a real word preceded by an unrelated real word and a real word preceded by a pseudoword. Facilitation effect was defined as a decrease in RT between a real word preceded by a related real word and a real word preceded by a pseudoword. Although the effect sizes of semantic priming did not differ between children and adults, the inhibition effect was larger and the facilitation effect was smaller in children. Repetition priming in children did not differ from that in adults. These findings challenge the idea of developmental invariance in implicit memory, suggesting that the strength of links between nodes in the semantic network increases and the range of spreading activation expands developmentally.