Metal-ion-induced expression of gene fragments from subseafloor micro-organisms in the Kumano forearc basin, Nankai Trough.

AIMS: Using substrate-induced gene-expression (SIGEX) screening on subseafloor sediment samples from the Nankai Trough, Japan, we identified gene fragments showing an induction response to metal ions. METHODS AND RESULTS: Environmental DNA libraries in Escherichia coli host cells were tested by the addition of metal ions (Ni2+ , Co2+ , Ga3+ or Mo6+ ), followed by cell sorting of clones exhibiting green fluorescence upon co-expression of green fluorescence protein downstream of the inserted gene fragments. One clone displayed Ni2+ -specific induction, three clones displayed Ga3+ -specific induction and three clones displayed an induction response to multiple metal ions. DNA sequence analysis showed that a variety of genes showed induction responses in the screened clones. CONCLUSIONS: Using the SIGEX approach, we retrieved gene fragments with no previously identified response to metal ions that exhibited metal-ion-induced expression. This method has the potential to promote exploration of gene function through gene-induction response. SIGNIFICANCE AND IMPACT OF THE STUDY: We successfully linked gene-induction response with sequence information for gene fragments of previously unknown function. The SIGEX-based approach exhibited the potential to identify genetic function in unknown gene pools from the deep subseafloor biosphere, as well as novel genetic components for future biotechnological applications.

Determination of HLA-A, -C, -B, -DRB1 allele and haplotype frequency in Japanese population based on family study.

The present study investigates the human leucocyte antigen (HLA) allele and haplotype frequencies in Japanese population. We carried out the frequency analysis in 5824 families living across Japanese archipelago. The studied population has mainly been typed for the purpose of transplant, especially the hematopoietic stem cell transplantation (HSCT). We determined HLA class I (A, B, and C) and HLA class II (DRB1) using Luminex technology. The haplotypes were directly counted by segregation. A total of 44 HLA-A, 29 HLA-C, 75 HLA-B, and 42 HLA-DRB1 alleles were identified. In the HLA haplotypes of A-C-B-DRB1 and C-B, the pattern of linkage disequilibrium peculiar to Japanese population has been confirmed. Moreover, the haplotype frequencies based on family study was compared with the frequencies estimated by maximum likelihood estimation (MLE), and the equivalent results were obtained. The allele and haplotype frequencies obtained in this study could be useful for anthropology, transplantation therapy, and disease association studies.

Validity, reliability and acceptability of the Japanese version of the General Well-Being Schedule (GWBS).

The General Well-Being Schedule (GWBS) is a widely used, self-administered questionnaire to measure subjective well-being. It contains 18 items and was originally hypothesized to have six domains; however, the following studies suggest other factorial models. We devised the schedule's Japanese edition following the standard procedure and tested its psychometric properties considering Japanese cultural characteristics. Subjects were 1224 middle-aged participants undergoing health checkups in Adachi, Tokyo. The reliability of the original version was high: alpha coefficient, 0.90; test-retest Pearson's correlation coefficient, 0.81; and intra-class correlation coefficient, 0.85. Confirmatory factor analyses supported the structural validity of the 6-factor, the following 3-factor and 4-factor models (goodness of fit index: 0.932, 0.946 and 0.937, respectively). A new 3-factor model developed after the exclusion of item 11, which had no correlation with the other variables, was proved to have a higher fitness index (0.958) than the other models. This 17-item version showed adequate reliability (alpha coefficient, 0.91; test-retest Pearson's correlation coefficient, 0.81; and intra-class correlation coefficient, 0.85) and concurrent validity compared with the General Health Questionnaire, State-Trait Anxiety Inventory, Center for Epidemiologic Studies Depression Scale, Self-rating Depression Scale and Profile Of Mood State (correlation coefficients: -0.75 to -0.53). The median time required to complete the schedule was five minutes. Completing this instrument was easy for 80% of the subjects. These findings attest to the adequacy of the 17-item Japanese GWBS for the comprehensive assessment of individual inner state.

[Validity of the values of maximal oxygen intake for women recommended in the 'Quantity of exercise for developing health' by Ministry of Health and Welfare].

The purpose of this study was to examine the value of maximal oxygen intake per kilogram body weight (VO2max/wt) recommended for women between 20 and 29 years of age by the Ministry of Public Welfare. The subjects were 199 female college students, who were not in the habit of drinking and smoking. Results obtained were as follows: After controlling the effects of age and percent body fat (%fat), VO2max/wt showed a significant correlations with %fat, high-density lipoprotein cholesterol and systolic blood pressure. The values of VO2max/wt corresponding to borderline values of these CHD risk factors ranged from 27.6 to 29.5ml/kg.min. The highest value of VO2max/wt was very close to the value recommended for women older than 60 years of age by the Ministry of Public Welfare. The subjects were then divided into 4 groups; at above and below the value of VO2max/wt (35ml/kg.min) recommended for women between 20 and 29 years of age, and at above and below the value of VO2max/wt (31ml/kg.min) recommended for women older than 60 years of age by the Ministry of Public Welfare. There were no significant differences among 4 groups in occurrence of abnormal values in CHD risk factors. The results of this study support the validity of the value of VO2max/wt (31ml/kg.min) recommended for women older than 60 years of age by the Ministry of Public Welfare. However, it seems that it is not necessary to recommend the value of VO2max/wt (35ml/kg.min) for women between 20 and 29 years of age, who were not in the habit of drinking and smoking.

Relationship of cigarette smoking to blood pressure and serum lipids and lipoproteins in men.

1. The relationship of cigarette smoking to blood pressure and serum lipids and lipoproteins was studied in 7608 men, ranging from 40 to 59 years of age. Analyses were performed separately for non-drinkers and drinkers. 2. After adjusting age and body mass index (BMI) in non-drinkers and age, BMI and alcohol intake in drinkers in forward stepwise multiple regression analysis, there was a dose-dependent negative relationship between cigarette smoking and diastolic blood pressure (DBP) and high density lipoprotein cholesterol (HDL-C), regardless of drinking habit. There was a dose-dependent positive relationship between cigarette smoking and the ratio of total cholesterol (TC) to HDL-C (TC:HDL-C) in non-drinkers, but not in drinkers. There was a dose-dependent negative relationship between cigarette smoking and TC and a positive relationship between cigarette smoking and triglycerides (TG) in drinkers, but not in non-drinkers. 3. After matching age and BMI in non-drinkers, subjects who smoked more than 30 cigarettes/day had significantly lower mean values of systolic blood pressure (SBP; 4.3%; P < 0.05), DBP (3.0%; P < 0.01) and HDL-C (15.5%; P < 0.01) and higher mean values of TC:HDL-C (25.0%; P < 0.01), TG (46.8%; P < 0.01) and beta-lipoprotein (12.0%; P < 0.01) than non-smokers. In drinkers, after matching age, BMI, and alcohol intake, subjects who smoked more than 30 cigarettes/day had significantly lower mean values of SBP (2.8%; P < 0.05), DBP (4.8%; P < 0.01), HDL-C (17.3%; P < 0.01) and TC (4.4%; P < 0.01) and higher mean values of TC:HDL-C (15.4%; P < 0.01) and TG (45.1%; P < 0.01) than non-smokers. 4. Although the results are somewhat variable, the present study reveals that cigarette smoking is negatively associated with SBP and DBP and unfavourably associated with lipids and lipoproteins, regardless of drinking habit.