Prediction and Validation of Metabolic Dysfunction-Associated Fatty Liver Disease Using Insulin Resistance-Related Indices in the Japanese Population.

Background: Recently, metabolic dysfunction-associated fatty liver disease (MAFLD), which is based on evidence of hepatic steatosis and any of the following three conditions: overweight/obesity, presence of type 2 diabetes mellitus, or evidence of metabolic dysregulation, has been proposed. It is uncertain how indices that predict insulin resistance (IR) are associated with MAFLD. Methods: Among subjects who had undergone health examinations at our hospital, 1,257 (787 men and 474 women) who underwent fatty liver evaluation, were included in this cross-sectional study. The discriminatory ability of each index for MAFLD was tested using a receiver operating characteristic curve analysis. The associations between the homeostasis model assessment-IR (HOMA-IR), triglyceride to high-density lipoprotein cholesterol (TG/HDL-C) ratio, triglyceride-glucose (TyG) index, and MAFLD were investigated using multiple logistic regression analysis. Results: The mean age and body mass index of MAFLD subjects were 58.1 years and 26.0 kg/m2 in men and 63.0 years and 26.1 kg/m2 in women, respectively. The cutoff values of HOMA-IR, TG/HDL-C ratio, and TyG index in men and women were 1.40 (area under the curve [AUC]: 0.81) and 1.90 (AUC: 0.88); 1.56 (AUC: 0.75) and 1.06 (AUC: 0.78); and 8.62 (AUC: 0.75) and 8.45 (AUC: 0.80), respectively. All indices were significantly higher in both men and women with MAFLD. In the multivariate models, the odds of MAFLD were higher among both men and women in the highest tertile than those in the lowest tertile. When subjects were divided by sex, presence of MAFLD, and alanine aminotransferase (ALT) values, all indices were significantly higher in both men and women with high ALT levels who were diagnosed with MAFLD. Conclusion: IR strongly correlated with MAFLD, particularly in subjects with high ALT levels.

Association of High-Density Lipoprotein Subclasses Levels with Sleep Duration and Other Lifestyles in Middle-Aged and Elderly Women: Cross-Sectional Study.

Objective: We aimed at investigating the association of high-density lipoprotein subclasses (HDL2-C and HDL3-C) levels with sleep duration, in comparison to other lifestyles in middle-aged and elderly women. Materials and Methods: A total of 69 women aged older than 40 who underwent "Anti-aging Health Checkups" were enrolled in the study. The analyses were conducted for all the subjects using personal data regarding clinical characteristics and lifestyle. Sleep duration was categorized into two groups of less than or more than 6 hrs. First, an analysis was performed to assess the correlation of two major HDL subclasses with various factors. Next, a multiple regression analysis was conducted to identify the association for each HDL2-C and HDL3-C with lifestyles such as sleep duration, daily breakfast, dinner time, habitual exercise, and drinking. Moreover, we examined the associations between HDL2-C and sleep duration combined with other lifestyle factors such as dinner time, daily breakfast, habitual exercise, and drinking. Results: In comparison to lifestyles, sleep duration had a strong association with only HDL2-C after adjustment for confounders. The "less 6 hrs sleep" group in combination with the "no exercise habit" or the "routine drinking habit" significantly decreased HDL2-C levels more than the assumed reference group. Regarding breakfast, there is a significant association between the "less than 6 hrs sleep with no daily breakfast" and the "more than 6 hrs sleep with daily breakfast." Conclusion: The results of this study may suggest that sufficient sleep might be significant for maintaining appropriate HDL2-C levels in middle-aged and elderly women under the condition that lifestyle might change during the ongoing COVID-19 pandemic.

Estimated Elovl6 and delta-5 desaturase activities might represent potential markers for insulin resistance in Japanese adults.

Purpose: Results from a recent study indicated that lower stearic acid/palmitic acid (SA/PA) and arachidonic acid/dihomo-γ-linolenic acid (AA/DGLA) ratios were associated with metabolically unhealthy obesity. However, this has not been extensively studied in the Japanese population. Methods: We recruited 291 Japanese subjects with serum free fatty acid profiles undergoing health examinations. Whole serum desaturase activity was estimated as the product: precursor ratio -SA/PA ratio for elongation of long-chain fatty acid family member 6 (Elovl6) and AA/DGLA for delta-5 desaturase (D5D). The determinants of Elovl6 and D5D activity were investigated using multiple regression analyses. Results: The Elovl6 and D5D activities exhibited a negative correlation with the logmatic-transformed TG/HDL-C ratio and TyG index. Multiple regression analyses revealed that the TG/HDL-C ratio and TyG index were negatively associated with Elovl6 and D5D activities. Most atherogenic markers were worse in the low Elovl6 or D5D activity group than in the high Elovl6 or D5D activity group. When study subjects were further stratified by TG levels, most atherogenic markers were the worst in the highest TG group in either the lowest Elovl6 or lowest D5D activity groups. Conclusion: The estimated Elovl6 and D5D activities might be useful markers of insulin resistance in Japanese subjects.

Oleic acid to stearic acid ratio might be a potential marker for insulin resistance in non-obese Japanese.

Results from a recent study indicate that a higher level of oleic acid/stearic acid ratio was associated with metabolically unhealthy obesity. This was further validated in cross-sectional and interventional studies; however, this was not extensively studied in a non-obese population. We recruited 260 Japanese subjects with serum free fatty acid profiles undergoing anti-aging health examinations. The determinants for oleic acid/stearic acid ratio were investigated using multiple regression analyses. To compare different markers, the subjects were classified based on oleic acid/stearic acid ratio and the combination of oleic acid/stearic acid ratio and triglyceride levels. The oleic acid/stearic acid ratio exhibited a positive correlation with the logmatic transformed triglyceride/high-density lipoprotein cholesterol ratio and the fasting triglycerides-glucose index, both of which were used as markers for insulin resistance. Multiple regression analyses revealed that the triglyceride/high-density lipoprotein cholesterol ratio and fasting triglyceride-glucose index were positively associated with the oleic acid/stearic acid ratio. Most markers were the worst in the highest triglyceride group in both oleic acid/stearic acid groups. In addition, most markers were worse in high oleic acid/stearic acid ratio group than low group. In conclusion, oleic acid/stearic acid ratio might be a useful marker for insulin resistance in non-obese Japanese subjects.

Usefulness of Triglyceride to High-Density Lipoprotein Ratio and Alanine Aminotransferase for Predicting Insulin Resistance and Metabolic Syndrome in the Japanese Population.

Background: Considering the fact that both alanine aminotransferase (ALT) and the triglyceride (TG)/high-density lipoprotein cholesterol (HDL-C) ratio are useful markers for cardiovascular disease, insulin resistance (IR), and metabolic syndrome (MetS), the possible association of ALT and TG/HDL-C ratio is conceivable, but the association is not well studied. Moreover, assessment of IR and MetS by a combination lipid and liver biomarkers has rarely been carried out. Methods: A total of 11,808 Japanese subjects who underwent an annual health examination and who were not taking medications were recruited. The association between TG/HDL-C ratio and ALT level was investigated using correlation and multiple linear regression analyses. Homeostasis model assessment of insulin resistance (HOMA-IR) and the number of MetS components were evaluated after subjects were stratified by TG/HDL-C ratio and ALT levels. Results: Correlation analysis revealed that ALT levels were positively associated with TG/HDL-C ratio. Moreover, multiple linear regression analysis revealed that TG/HDL-C ratio was positively associated with ALT levels, and the association was attenuated but persisted in both men and women after additional adjustment for visceral adiposity (body mass index, waist circumference, or fatty liver) and IR (HOMA-IR). HOMA-IR and MetS component number increased with either ALT or TG/HDL-C ratio in both men and women. HOMA-IR and MetS component number were particularly high (HOMA-IR: men 2.82, women 3.22, MetS component number: men 2.6, women 2.1) in subjects with highest ALT level and TG/HDL-C ratio. Conclusions: TG/HDL-C ratio showed a positive association with ALT levels in both sexes. Assessments of IR and MetS can become more precise by evaluating TG/HDL-C ratio and ALT, simultaneously.

Relationship between Serum Antioxidative Vitamin Concentrations and Type 2 Diabetes in Japanese Subjects.

A high intake of green leafy vegetables rich in antioxidative nutrients such as vitamin C and ß-carotene may protect against the risk of type 2 diabetes. Measurement of the circulating nutrient concentrations can indicate the nutrient status more directly, and vitamin C and carotenoids are recognized as good biomarkers for the intake of fruits and vegetables. The aim of this study was to investigate the relationships between serum antioxidative vitamin concentrations and type 2 diabetes in Japanese subjects. The study subjects comprised 506 men and 493 women who first underwent anti-aging health checks at Tokai University Tokyo Hospital. Serum concentration of vitamin (V) A, VC, α-tocoferol, ß-carotene, VB12, folate, ferritin and homocysteine, and fasting plasma glucose and HbA1c were used for analysis. Low levels of ß-carotene and VC were significantly associated with dysglycemia. Diabetic subjects showed significantly decreased ß-carotene and VC levels, and multivariate analyses suggested that low levels of ß-carotene and VC were factors related to diabetes. Low levels of ß-carotene and VC are significantly related to dysglycemia/type 2 diabetes, and encouraging people at a higher risk of diabetes to take more green vegetables may be useful as a dietary intervention to improve the antioxidative vitamin status and dysglycemia.

Factors associated with serum total homocysteine level in type 2 diabetes.

OBJECTIVES: The aim of this study was to investigate the determinants of serum total homocysteine level (tHcy) in patients with type 2 diabetes mellitus (DM) according to sex. METHODS: A total of 1,276 Japanese, diabetics (n = 280) with a control group of non-diabetics (n = 996), were enrolled into the study from 2003 to 2005. This cross-sectional study was conducted for all the subjects, using personal data regarding clinical characteristics and lifestyle. Multiple regression analysis was performed to analyze the association of tHcy with selected factors. RESULTS: In diabetic subjects, estimated glomerular filtration rate (eGFR) and serum creatinine levels (Cre), even those within the normal range, were strongly associated with tHcy after adjustment in both sexes; the standardized partial regression coefficient of eGFR for tHcy was -0.251, (p = 0.001) in diabetic men and -0.523, (p < 0.001) in diabetic women. Furthermore, the eGFR of the diabetics, except patients with nephropathy, also had significant association with tHcy in both sexes. Fasting plasma glucose levels and serum triglyceride levels were strongly associated with tHcy in diabetic men only. HbA1c was also associated with tHcy in diabetic men only, though not as significantly. Age and presence of hypertension were significantly associated with tHcy in women. CONCLUSIONS: This study suggests that there are some differences in the factors associated with tHcy between diabetics and non-diabetics, and between the sexes. There is, therefore, circumstantial evidence that elevated tHcy should be evaluated clinically. Because tHcy was strongly associated with eGFR and Cre, even within the normal ranges, tHcy may have important implications regarding the microangiopathy of the kidney and atherosclerosis.

NADH dehydrogenase subunit-2 237 Leu/Met polymorphism modifies the effects of alcohol consumption on risk for hypertension in middle-aged Japanese men.

NADH dehydrogenase subunit-2 237 leucine/methionine (ND2-237 Leu/Met) polymorphism is associated with longevity in the Japanese population, and the ND2-237Met genotype may exert antiatherogenic effects. To investigate whether ND2-237 Leu/Met polymorphism is associated with risk of hypertension, we conducted a cross-sectional study of 398 Japanese male subjects. The frequency of hypertension was significantly higher in ND2-237Leu genotypic men than in ND2-237Met genotypic men. On analysis of covariance, the interaction between ND2-237 Leu/Met polymorphism and habitual drinking was significantly associated with both systolic blood pressure and diastolic blood pressure. Multiple logistic regression analysis revealed that the ND2-237Met genotype, particularly in younger subjects (age <60 years), had a lower odds ratio for hypertension than the ND2-237Leu genotype. Moreover, the association of ND2-237 Leu/Met polymorphism with hypertension may depend on the frequency of alcohol consumption. The odds ratio for hypertension was significantly higher in daily drinkers with ND2-237Leu when compared with non- or ex-drinkers with ND2-237Leu. However, the association between the ND2-237Met genotype and hypertension may not depend on the frequency of alcohol consumption. The present results suggest that ND2-237 Leu/Met polymorphism is associated with hypertension and that modification of hypertension risk is dependent on alcohol consumption in middle-aged Japanese men.

Longevity-associated NADH dehydrogenase subunit-2 237 Leu/Met polymorphism influences the effects of alcohol consumption on serum uric acid levels in nonobese Japanese men.

NADH dehydrogenase subunit-2 237 leucine/methionine (ND2-237 Leu/Met) polymorphism is reportedly associated with longevity in the Japanese population. The ND2-237Met genotype may confer resistance to cardiovascular and cerebrovascular atherogenic diseases. Hyperuricemia is one of the risk factors for cardiovascular disease. To investigate whether ND2-237 Leu/Met polymorphism is associated with serum uric acid (SUA) levels, we conducted a cross-sectional study in 321 healthy Japanese male subjects. In nonobese (body mass index, BMI<25) male subjects, interaction between ND2-237 Leu/Met genotypes and drinking frequency on SUA levels was observed (P=0.031). The SUA levels were significantly higher in daily drinkers with ND2-237Leu than in non-daily drinkers with ND2-237Leu (P=0.018). In nonobese men, after adjustment for covariates, daily drinkers with ND2-237Leu had a significantly higher odds ratio (OR) for hyperuricemia (SUA> or =6.5 mg/dl: vs. daily drinkers with ND2-237Met, OR=3.26, 95% confidence interval (CI) 1.14-9.29; vs. non-daily drinkers with ND2-237Leu, OR=3.22, 95% CI 1.39-7.45; SUA> or =7.0 mg/dl: vs. non-daily drinkers with ND2-237Met, OR=3.53, 95% CI 1.00-12.4). However, in obese (BMI> or =25) men, no significant interaction between ND2-237 Leu/Met polymorphism and habitual drinking on SUA levels or on the risk for hyperuricemia was observed. These results suggest that ND2-237 Leu/Met polymorphism modulates the effects of daily alcohol consumption on SUA levels in nonobese Japanese men.

Personal features and dropout from diabetic care.

OBJECTIVE: To investigate the personal features associated with dropout from regular outpatient care among persons with type 2 diabetes mellitus (DM). METHODS: A total of 160 DM patients were enrolled in the study. As a retrospective analysis, outpatient's clinical characteristics, lifestyle, or social features were gathered from their medical records or interview sheets. All the subjects were divided into two groups by adherence to diabetic care, namely, 'dropout case' (DC), or 'ongoing case' (OC), and were subjected to comparative analysis. We called the patients who did not receive outpatient treatment from the clinic on a regular basis, including treatment from other clinics or dropout of diabetic care, as DC. In contrast, patients who regularly visited the clinic were defined as OC. An unconditional multiple logistic regression analysis was performed to analyze the association of a dherence to diabetic care with several personal features. RESULTS: Sixty-eight of 160 subjects (42.5%) were recognized as DC. The remaining 92 subjects (57.5%) were considered as OC. Young age (p=0.045), low plasma glucose (p=0.005) and hemoglobin A1c (HbA1c) levels (p=0.005), nonmedication (p<0.001) and no past history of DM (p=0.007) at the initial visit were the features related to dropout by crude analysis. Even after adjustment for age and gender by multivariate analysis, there remained significant inverse associations of dropout with HbA1c level, medical treatment (oral agents or insulin) and previous DM history. Neither occupation, distance from residence to clinic, smoking habit nor drinking habit was associated with dropout. Dropout mostly occurred after the initial or second visit. CONCLUSIONS: A mild condition of DM may be related to dropout from regular outpatient care. It may be necessary to clearly show the objectives and importance of regular visit to an outpatient clinic for diabetic care, particularly for screened mild DM cases in public health activities.

Longevity-associated mitochondrial DNA 5178 C/A polymorphism is associated with fasting plasma glucose levels and glucose tolerance in Japanese men.

Mitochondrial DNA 5178 cytosine/adenine (Mt5178 C/A) polymorphism is reportedly associated with longevity in the Japanese population, and the Mt5178A genotype may resist the onset of type 2 diabetes. To investigate whether Mt5178 C/A polymorphism is associated with glucose tolerance, we conducted a cross-sectional study using the 75-g oral glucose tolerance test (OGTT) in which non-diabetic Japanese male subjects were classified into three subgroups by body mass index (BMI): BMI<22 (n=91); 22< or =BMI<25 (n=138); and BMI> or =25 (n=67). The frequency of Mt5178A was significantly lower among 'BMI<22' subjects exhibiting impaired fasting glucose and impaired glucose tolerance than among those with normal glucose tolerance. In the 'BMI<22' group, fasting plasma glucose (FPG) levels and plasma glucose levels at 60 and 120 min after glucose load (OGTT-1h and OGTT-2h, respectively) were significantly lower in the Mt5178A genotype than in the Mt5178C genotype. After adjusting for age, BMI, habitual smoking, habitual drinking and family history of diabetes, FPG levels and OGTT-2h levels were still significantly lower in the Mt5178A genotype than in the Mt5178C genotype. However, after adjusting for covariates, in both the '22< or =BMI<25' and 'BMI> or =25' groups, FPG levels were significantly higher in the Mt5178A genotype than in the Mt5178C genotype. Differences in the effect of alcohol consumption on FPG levels and glucose tolerance between the Mt5178 C/A genotypes were observed. The present results suggest that Mt5178 C/A polymorphism may be associated with FPG levels and glucose tolerance in middle-aged Japanese men.

Longevity-associated NADH dehydrogenase subunit-2 polymorphism and serum electrolyte levels in middle-aged obese Japanese men.

Mitochondrial DNA 5178 cytosine/adenine polymorphism, which is also called NADH dehydrogenase subunit-2 237 leucine/methionine (ND2-237 Leu/Met) polymorphism is associated with Japanese longevity. This polymorphism is widely associated with blood pressure, serum lipid levels, hematological parameters, intraocular pressure, and serum protein fraction levels. However, there have been no reports on the association between ND2-237 Leu/Met polymorphism and serum electrolyte levels. To investigate this relationship, we performed an association study in 321 healthy middle-aged Japanese men. Crude data showed that serum sodium levels and serum chloride levels were significantly lower in men with ND2-237 Met than in those with ND2-237 Leu (P = 0.021 and 0.003, respectively). Cigarette consumption and body mass index were significantly and positively associated with serum chloride levels (P = 0.002 and 0.008, respectively) and hemoglobin levels were significantly and negatively associated with them (P = 0.007) in ND2-237 Leu genotypic men. In men with ND2-237 Met, only hemoglobin levels were significantly and negatively associated with serum chloride levels (P = 0.025). After adjusting for covariates, only in male obese (body mass index> or =25) subjects, serum sodium and chloride levels remained significantly lower, and serum calcium levels appeared to be significantly higher in ND2-237 Met than in ND2-237 Leu (P = 0.013, <0.001, and 0.046, respectively). Longevity-associated NADH dehydrogenase subunit-2 polymorphism may influence serum electrolyte levels in middle-aged obese Japanese men.

Interaction between longevity-associated mitochondrial DNA 5178 C/A polymorphism and cigarette smoking on hematological parameters in Japanese men.

Mitochondrial DNA 5178 C/A (mt5178 C/A), namely NADH dehydrogenase subunit 2 237 Leu/Met, polymorphism is as reported in literature associated with longevity and susceptibility to ischemic heart disease or cerebrovascular disorders in the Japanese population. Previous reports suggested that mt5178A genotype exerts antiatherogenic effects. The aim of this study was to investigate whether mt5178 C/A polymorphism is associated with hematological parameters, such as thrombogenic risk factors for myocardial infarction and stroke, in 321 healthy Japanese men. No significant differences were observed between mt5178 C/A genotypes, but in subjects with body mass index (BMI) of < or = 23, this polymorphism influenced the effects of habitual smoking on hematological parameters. Red blood cell (RBC) counts were significantly lower and mean corpuscular hemoglobin (MCH) levels were significantly higher in smokers with mt5178A than nonsmokers with mt5178A. Platelet counts were significantly higher in smokers with mt5178C than nonsmokers with mt5178C. Cigarette consumption was strongly associated with RBC counts, mean corpuscular volume levels, and MCH levels for men with mt5178A, and was associated with platelet counts for those with mt5178C. Moreover, BMI was significantly positively associated with RBC counts and platelet counts only in men with mt5178A, age was significantly negatively associated with RBC counts only in men with mt5178C. These data suggest that mt5178 C/A polymorphism may influence the effects of cigarette smoking on hematological parameters in healthy BMI < or = 23 Japanese men.

Genotoxicity of 2-[2-(acetylamino)-4-[bis(2-hydroxyethyl)amino]-5-methoxyphenyl]-5-amino-7-bromo-4-chloro-2H-benzotriazole (PBTA-6) and 4-amino-3,3'-dichloro-5,4'-dinitro-biphenyl (ADDB) in goldfish (Carassius auratus) using the micronucleus test and the comet assay.

2-[2-(Acetylamino)-4-[bis(2-hydroxyethyl)amino]-5-methoxyphenyl]-5-amino-7-bromo-4-chloro-2H-benzotriazole (PBTA-6) and 4-amino-3,3'-dichloro-5,4'-dinitrobiphenyl (ADDB) are two compounds, which show strong mutagenicity toward bacteria, that have been identified as major mutagens in river water in Japan. In the present study, we examined the genotoxicity of PBTA-6 and ADDB in goldfish (Carassius auratus) by the micronucleus test and single-cell gel electrophoresis (comet assay). The frequencies of micronuclei in gill cells gradually increased until 96h after i.p. injection of PBTA-6 and ADDB at doses of 50mg/kg body weight, and then decreased 144h after injection. PBTA-6 induced micronuclei in gill cells dose-dependently at a dose range of 1-100mg/kg body weight, giving significantly high frequencies at doses of 50 and 100mg/kg body weight. On the other hand, no significant increase was observed in the peripheral erythrocytes of goldfish exposed to PBTA-6 or ADDB. In the comet assay, values of DNA tail moment and tail length in peripheral erythrocytes increased significantly until 6h after the i.p. injection of PBTA-6 (50mg/kg body weight), only to decrease by 9h after injection. Both the DNA tail moment and tail length were dose-dependently increased by injections of PBTA-6 at doses ranging from 1 to 50mg/kg. Significantly high values for tail moment and tail length were found in peripheral erythrocytes 3h after an i.p. injection of ADDB and persisted for up to 6h. These results show that both PBTA-6 and ADDB have genotoxic effects in goldfish.

Longevity-associated mitochondrial DNA 5178 A/C polymorphism is associated with intraocular pressure in Japanese men.

BACKGROUND: Mitochondrial DNA 5178 adenine/cytosine (mt5178 A/C) polymorphism has been reported to be associated with longevity in Japanese individuals, with experimental findings concluding that mt5178 A is an anti-atherogenic genotype. The aim of this study was to determine whether mt5178 A/C polymorphism influences intraocular pressure (IOP), and its relationship with the development of arteriosclerosis. METHODS: Four hundred and forty-nine male volunteers who had visited a general hospital for medical check-up between August 1999 and August 2000 were enrolled. Of these, 386 Japanese men who had not undergone any medical treatment for hypertension, ocular hypertension or glaucoma were selected as subjects of this study. After these subjects were genotyped, a cross-sectional study regarding the relationship between genotype and IOP was conducted. RESULTS: Mean IOP was significantly higher in men with mt5178 C (13.3 mmHg) than in those with mt5178 A (12.7 mmHg) (P = 0.037). This significant difference in mean IOP between the two genotypes remained evident after adjusting for age, body mass index, blood pressure, habitual smoking and habitual drinking. Interactions between mt5178 A/C polymorphism and habitual smoking or daily alcohol consumption with regard to IOP were observed. According to multiple regression analysis, habitual smoking was significantly associated with IOP in men with mt5178 A (P for trend = 0.020), while daily alcohol consumption was significantly associated with IOP in those with mt5178 C (P for trend = 0.021). CONCLUSION: Longevity-associated mitochondrial DNA 5178 A/C polymorphism may be associated with IOP in Japanese men.