Mikiko Kobayashi-Miura

Association analysis of the NOD2 gene with susceptibility to graft-versus-host disease in a Japanese population

Members of the nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) family participate in the innate immune system, exerting widespread effects on cytokine secretion, autophagy, and apoptosis. Recent studies in Caucasians revealed the association between mutants of NOD2, a member of the NLR family, and severity of acute graft-versus-host disease (GVHD). NOD2 polymorphism screening has been recommended for donor selection and risk assessment at bone marrow transplantation. To investigate whether NOD2 plays a role in the pathogenesis of GVHD in a Japanese population, we examined DNA from 142 bone marrow transplant patient/donor pairs to detect genetic variation in the NOD2 gene. No genetic variants of NOD2 were associated with the severity of acute GVHD in our patients. However, a weak association between a single nucleotide polymorphism in the NOD2 gene (R471C) and acute myeloid leukemia in the bone marrow patients (p = 0.029, odds ratio 4.08, 95% CI 1.22–13.67) was detected. This polymorphism was not prevalent in 479 Crohn’s disease (CD) patients in Japan. These results suggest that, in the Japanese population, unlike the Caucasian, NOD2 is not a major contributor to susceptibility to severe acute GVHD.

GM-CSF-Independent CD1a Expression in Epidermal Langerhans Cells: Evidence from Human CD1A Genome-Transgenic Mice

the primer-binding regions, our present study might underestimate the frequencies detectable in the skin cancer samples. Nevertheless, the copy number of the viruses per human genome within the positive tested samples was very low. In the representative examples depicted in Figure 1c, HPyV6 showed much higher cycle thresholds than the cellular gene control thyroid peroxidase, which is normally unaffected by gains and losses in MCC and thus should represent a curve for two copies per cell (Paulson et al., 2009). The more frequent presence of HPyV6 compared with HPyV7 among the skin tumor samples is in line with the serological data available for these viruses (Schowalter et al., 2010). The observations of a similar detection rate of HPyV6 and HPyV7 on the skin surface of healthy volunteers, the higher seropositivity for HPyV6, and an increased presence of HPyV6 in skin tumor samples are suggestive for either a broader cell tropism, a higher survival capability within the skin, or a more efficient skin entry mechanism of HPyV6. The low viral load presence of HPyV6 and HPyV7 in some of the cancer samples reflects the situation of MCV, which can also be detected at low quantities in different tissue specimens irrespective of the pathological status (Loyo et al., 2010). In summary, the low level presence of HPyV6 and HPyV7 within the 108 analyzed skin cancer samples, i.e., clearly lower than one viral copy per cell, does not deliver any evidence for a significant role of these polyomaviruses for the pathology of the respective skin cancers, but it certainly exclude a role of these viruses in the maintenance of the malignant phenotype of the analyzed skin cancers.

Association Between Feeling Upon Awakening and Use of Information Technology Devices in Japanese Children

Background The objective of this study was to clarify the relationship between feeling upon awakening (FA) and time spent using information technology (IT) devices by children in kindergartens, elementary schools, and junior high schools in Shimane, Japan. Methods In October 2008, a self-report survey was distributed to 2075 children in kindergartens (n = 261), elementary schools (n = 1162), and junior high schools (n = 652) in Shimane, Japan. The questionnaire gathered data on sex, school year, feeling upon awakening, and time spent using IT devices after school (television, videos on television, video games, personal computers, and cellular phones). After adjusting for sex and school year, data were analyzed by multivariate logistic regression analysis to calculate odds ratios (ORs) and 95% confidence intervals (CIs). Results A total of 2030 children completed this survey (response rate, 97.8%). Negative FA was associated with watching television more than 2 hours/day (OR = 1.51, 95% CI = 1.23–1.85), playing video games more than 30 minutes/day (1.50, 1.20–1.87), and using personal computers more than 30 minutes/day (1.35, 1.04–1.75). Conclusions Time spent using IT devices affected the FA of children in kindergarten through junior high school. We propose the development of guidelines regarding the appropriate amount of time this population should spend using IT devices.

Association of plasma free amino acids with hyperuricemia in relation to diabetes mellitus, dyslipidemia, hypertension and metabolic syndrome

Previous studies demonstrated independent contributions of plasma free amino acids (PFAAs) and high uric acid (UA) concentrations to increased risks of lifestyle-related diseases (LSRDs), but the important associations between these factors and LSRDs remain unknown. We quantified PFAAs and UA amongst Japanese subjects without LSRDs (no-LSRD, n = 2805), and with diabetes mellitus (DM, n = 415), dyslipidemia (n = 3207), hypertension (n = 2736) and metabolic syndrome (MetS, n = 717). The concentrations of most amino acids differed significantly between the subjects with and without hyperuricemia (HU) and also between the no-LSRD and LSRD groups (p < 0.05 to 0.001). After adjustment, the logistic regression analyses revealed that lysine in DM, alanine, proline and tyrosine in dyslipidemia, histidine, lysine and ornithine in hypertension, and lysine and tyrosine in MetS demonstrated significant positive associations with HU among the patients with LSRDs only (p < 0.05 to 0.005). By contrast, arginine, asparagine and threonine showed significant inverse associations with HU in the no-LSRD group only (p < 0.05 to 0.01). For the first time, we provide evidence for distinct patterns of association between PFAAs and HU in LSRDs, and postulate the possibility of interplay between PFAAs and UA in their pathophysiology.

Alteration in plasma free amino acid levels and its association with gout

BackgroundStudies on the association of plasma-free amino acids with gout are very limited and produced conflicting results. Therefore, we sought to explore and characterize the plasma-free amino acid (PFAA) profile in patients with gout and evaluate its association with the latter.MethodsData from a total of 819 subjects (including 34 patients with gout) undergoing an annual health examination program in Shimane, Japan were considered for this study. Venous blood samples were collected from the subjects and concentrations of 19 plasma amino acids were determined by high-performance liquid chromatography–electrospray ionization–mass spectrometry. Student’s t-test was applied for comparison of variables between patient and control groups. The relationships between the presence or absence of gout and individual amino acids were investigated by logistic regression analysis controlling for the effects of potential demographic confounders.ResultsAmong 19 amino acids, the levels of 10 amino acids (alanine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, serine, tryptophan, valine) differed significantly (P < .001 to .05) between the patient and control groups. Univariate logistic regression analysis revealed that plasma levels of alanine, isoleucine, leucine, phenylalanine, tryptophan and valine had significant positive associations (P < .005 to .05) whereas glycine and serine had significant inverse association (P < .05) with gout.ConclusionsThe observed significant changes in PFAA profiles may have important implications for improving our understanding of pathophysiology, diagnosis and prevention of gout. The findings of this study need further confirmation in future large-scale studies involving a larger number of patients with gout.

Complete solubilization of cartilage using the heat-stable protease thermolysin for comprehensive GAG analysis

Articular cartilage comprises collagens, proteoglycans, and glycosaminoglycans (GAGs) together with water, in hyaline matrixes. Articular cartilage is resistant to proteolytic solubilization for comprehensive GAG analyses partly because of assemblies of collagen fibers with thermolabile hydrogen bonds. In this study, we used the heat-stable protease thermolysin to digest collagen in solid articular cartilage at 70 °C and compared the efficiencies of collagen digestion and GAG extraction to those with collagenase digestion at 50 °C. Overnight digestion with thermolysin completely solubilized cartilage, whereas collagenase with >10-times higher proteolytic activity digested <20% of collagen. Following thermolysin treatments, almost all GAGs were extracted from the cartilage, whereas only 56% of GAGs were extracted after collagenase digestion. Disaccharide analyses of extracted GAG chains revealed >98% extraction efficiencies of several GAG classes from thermolysin-treated cartilage, compared with <60% extraction efficiencies using collagenase, depending on GAG classes. These results indicate that thermolysin allows complete GAG extraction from solid articular cartilage and that complete solubilization is required for accurate and reproducible analyses of cartilage GAGs. Hence, thermolysin offers a tool for complete solubilization of cartilage prior to comprehensive GAGomic analysis, and is likely applicable to other collagen-rich tissues such as ligaments, skin, and blood vessels.

Quantitative analyses of rat articular cartilage matrix during the maturation

among which RGA, REPRESSOR OF GA1-3, is commonly regarded as the most vital one. In the transgenic plants carrying pER:RGAΔ17-HA expressing the gain-of-function of RGA in a estradiol-inducible manner, the embryo showed the similar phenotype with the one sprayed with PAC. On the contrary, the embryo in della quandruple mutant rga gai rgl1 rgl2 displayed larger size, mimicking the one with exogenous application of GA3, which further proved the important effects imposed by gibberellin in this process. Seed embryogenesis is followed by storage reserve accumulation, desiccation and dormancy. Surprisingly, gibberellins function as a seemingly negative force in the later phase. Both previous research and our experiment data showed that seed storage protein and oil content decreased in the della mutant with reduced seed dormancy or even vivpary phenotype, demonstrating the possibility that gibberellins are also involved in the seed maturation stage. In summary, gibberellin may functions both in the early and later periods of seed development.