Background and Objectives: The gut microbiota is associated with human health and dietary nutrition. Various studies have been reported in this regard, but it is difficult to clearly analyze human gut microbiota as individual differences are significant. The causes of these individual differences in intestinal microflora are genetic and/or environmental. In this study, we focused on differences between identical twins in Japan to clarify the effects of nutrients consumed on the entire gut microbiome, while excluding genetic differences. Materials and Methods: We selected healthy Japanese monozygotic twins for the study and confirmed their zygosity by matching 15 short tandem repeat loci. Their fecal samples were subjected to 16S rRNA sequencing and bioinformatics analyses to identify and compare the fluctuations in intestinal bacteria. Results: We identified 12 genera sensitive to environmental factors, and found that Lactobacillus was relatively unaffected by environmental factors. Moreover, we identified protein, fat, and some nutrient intake that can affect 12 genera, which have been identified to be more sensitive to environmental factors. Among the 12 genera, Bacteroides had a positive correlation with retinol equivalent intake (rs = 0.38), Lachnospira had a significantly negative correlation with protein, sodium, iron, vitamin D, vitamin B6, and vitamin B12 intake (rs = −0.38, −0.41, −0.39, −0.63, −0.42, −0.49, respectively), Lachnospiraceae ND3007 group had a positive correlation with fat intake (rs = 0.39), and Lachnospiraceae UCG-008 group had a negative correlation with the saturated fatty acid intake (rs = −0.45). Conclusions: Our study is the first to focus on the relationship between human gut microbiota and nutrient intake using samples from Japanese twins to exclude the effects of genetic factors. These findings will broaden our understanding of the more intuitive relationship between nutrient intake and the gut microbiota and can be a useful basis for finding useful biomarkers that contribute to human health.

DNA methylation levels will be important for detection of epigenetic effects. However, there are few reports showing sex-related differences in the sensitivity to DNA methylation. To evaluate their sex-related individual differences in the sensitivity to methylation rigorously, we performed a systematic analysis of DNA methylation in monozygotic twins, an optimal model to evaluate them because the genetic backgrounds are the same. We examined 30 male and 43 female older monozygotic twin pairs recruited from the registry established by the Center for Twin Research, Osaka University. Their methylation levels were determined using the Infinium HumanMethylation450 BeadChip Kit (Illumina), which interrogated 485577 highly informative CpG sites at the single-nucleotide resolution, and the median methylation level was calculated for each of the 25657 CpG islands. Within-pair differences of methylation levels (WPDMs) were greater in male pairs than female pairs for 86.0 % of autosomal CpG islands, but were higher in female pairs than male pairs for 76.7 % of X chromosomal CpG islands. Mean WPDMs of CpG islands in each autosomal chromosome were significantly higher in male pairs than in female whereas that in X chromosome was significantly higher in female pairs than in male. Multiple comparison indicated that WPDMs in three autosomal and two X-chromosomal CpG islands were significantly greater in male pairs, whereas those in 22 X-chromosomal CpG islands were significantly greater in female pairs. Sex-related differences were present in the WPDMs of CpG islands in individuals with the same genetic background. These differences may be associated with the sexual influences in susceptibility of some diseases.

Background: Epigenetic factors associated with the development of autoimmune diseases are unclear. Monozygotic twin pairs discordant for positive antithyroglobulin autoantibodies (TgAb) are useful to examine the epigenetic factors because of their identical genetic background. This study aimed to clarify the discordant epigenetic differences affecting the development of TgAb. Methods: Subjects were selected from 257 Japanese monozygotic twins, recruited from the registry established by the Center for Twin Research at Osaka University. TgAb positive concordant (PC) pairs were 5.7% (four pairs) and 9.6% (18 pairs) of male and female pairs, respectively. TgAb discordant (DC) pairs were 11.4% (eight pairs) and 8.0% (15 pairs) of male and female pairs, respectively. TgAb negative concordant (NC) pairs were 78.6% (55 pairs) of male pairs and 74.3% (139 pairs) of female pairs. To perform stricter grouping, the cut-off value for positive TgAb was set to 50.0 IU/mL (TgAb negative: <28.0 IU/mL; TgAb positive: ≥50.0 IU/mL; TgAb borderline: ≥28.0 IU/mL and <50.0 IU/mL). Nineteen discordant (6 male and 13 female pairs) and 185 concordant pairs (48 male and 137 female pairs) for TgAb positivity were finally examined. DNA methylation levels of genomic DNA were evaluated using the Infinium HumanMethylation450 BeadChip Kit. Gene polymorphisms were also genotyped using the Omni5-4 BeadChip Kit to clarify genetic background specific for discordant twins. Results: No CpG sites were found with significant within-pair differences of methylation levels in TgAb DC pairs after correction for multiple comparisons. However, 155 polymorphisms specific for TgAb DC pairs were significantly different in genotype frequencies from those of concordant pairs, and none of them were located on the HLA region of chromosome 6. In TgAb DC pairs with some specific genotypes of these polymorphisms, four CpG sites were observed exhibiting significant within-pair differences in each DC pair, even after correction for multiple comparisons. Conclusions: This study found that the genetic background specific for TgAb DC twins who are susceptible to epigenetic changes are different from that specific for TgAb PC twins, and it clarified the genotype-based epigenetic differences in TgAb DC monozygotic twins.

The number of rubella cases has increased in Japan, especially among adults. Rubella infection in pregnant females can lead to congenital rubella syndrome (CRS). The Japanese government is promoting vaccination to prevent CRS. This study performs a cost-effectiveness analysis of the following four methods: (1) females who wished to become pregnant and partners, with an antibody-titer test; (2) females only, with an antibody-titer test; (3) females and males, without an antibody-titer test; (4) females only, without an antibody-titer test. A decision tree model with a hypothetical cohort of 500 males and 500 females was used for the analysis, and the parameters were obtained from previous studies. The number of avoidances of CRS was defined as the effect. Compared to the case where methods were not implemented, the number of CRS cases that can be prevented by implementing the methods was 0.0115589 by (1) and (3) and 0.0147891 by (2) and (4). The cost effectiveness of (1) to (4) was 287,413,677 JPY, 135,050,529 JPY, 388,524,974 JPY, and 197,744,219 JPY, respectively (1 JPY = 0.00963247 USD). Method (2) was the most cost-effective and did not change by sensitivity analysis. We conclude that the vaccination for females only with an antibody-titer test is recommended.

Bioelectrical impedance analysis (BIA)-derived phase angle (PhA) is a valuable parameter to assess physical health. However, the genetic and environmental aspects of PhA are not yet well understood. The present study aimed to estimate the heritability of PhA and investigate the relationships between PhA and anthropometric measurements. PhA and skeletal muscle mass index (SMI) were examined using multi-frequency BIA in 168 Japanese twin volunteers (54 males and 114 females; mean age = 61.0 ± 16.5 years). We estimated the narrow-sense heritability of these parameters and the genetic and environmental relationships between them using a genetic twin modeling. For the PhA, 51% (95% confidence interval: 0.33, 0.64) of the variance was explained by additive genetic effects, and 49% (95% confidence interval: 0.36, 0.67) was explained by unique environmental effects. The heritability of PhA was lower than the height, body weight, and body mass index. PhA shared almost no genetic variation with anthropometric measurements and SMI but shared an environmental variation (14%) with SMI. These findings suggest that the genes affecting PhA are different than those affecting anthropometric measurements and SMI. The correlation between PhA and SMI is caused by common environmental factors.