Vitamin B complex exerts its function in energy metabolism, immune function and DNA synthesis, methylation and repair(Reference Mikkelsen and Apostolopoulos1). Vitamin B deficiency has been associated with cardiovascular disorders, particularly in ageing population(Reference Mikkelsen and Apostolopoulos1).
Among the B complex group, vitamin B1 and vitamin B3 come mainly from cereals, beef and pork, seeds and nuts, and yeast(Reference Suzuki, Goto and Ota2). Vitamin B1 deficiency results in beriberi, a neurological and cardiovascular disorder(Reference Eshak and Arafa3), while deficient vitamin B3 can cause pellagra-induced dilated cardiomyopathy(Reference Crook4). The potential positive impacts of vitamin B1 and B3 on cardiovascular health were suggested in several animal studies and supplemental clinical trials. In a Langendorff perfused rat hearts, vitamin B1 excreted protective effects against myocardial ischaemic injury via maintaining mitochondrial size and ATP levels(Reference Yamada, Kusakari and Akaoka5). A randomised controlled trial on chronic heart failure patients who used diuretics reported that 300 mg/d of vitamin B1 supplementation for 28 d increased the left ventricular ejection fraction by 3·9 %(Reference Schoenenberger, Schoenenberger-Berzins and der Maur6). On the other hand, 1500–2000 mg/d vitamin B3 supplementation was shown to decrease LDL-cholesterol, TAG and lipoprotein(a) levels, while increasing HDL-cholesterol level(Reference McKenney7,Reference Linder, Willmann and Kantartzis8) . A meta-analysis of clinical trials suggested that vitamin B3 supplements significantly reduced major coronary events, stroke and other cardiovascular events(Reference Bruckert, Labreuche and Amarenco9).
Despite the abundant evidence on cardiovascular beneficial effects of vitamins B1 and B3 from animal studies(Reference Eshak and Arafa3,Reference Wexler and Lutmer10,Reference Freye11) and supplemental clinical trials(Reference Canner, Berge and Wenger12,Reference Schandelmaier, Briel and Saccilotto13) , no human observational studies so far have investigated the associations of dietary vitamins B1 and B3 intakes with risk of CVD. Previous studies indicated that food sources rich in vitamins B1 and B3 such as fish/seafood and vegetables were associated with a reduced risk of mortality from CVD(Reference Yamagishi, Iso and Date14,Reference Nagura, Iso and Watanabe15) . Yet, the evidence on dietary vitamin B complex/CVD association was mainly directed to vitamins B2, B6, B12 and folate, while the effects of dietary vitamins B1 and B3 intakes were not studied. Another issue is that most of the clinical trials on cardiovascular risk used high dosages of vitamin B1 (200–300 mg/d) and B3 supplement (1500–2000 mg/d), while the RDA of vitamin B1 for Japanese men and women aged 50–69 years were 1·3 and 1·1 mg/d, respectively, and RDA of vitamin B3 were 14 and 11 mg/d, respectively(Reference Shibata, Fukuwatari and Imai16). Among Japanese men aged 30–49 years, the estimated average requirement and RDA were 1·2 and 1·4 mg/d, respectively, for vitamin B1, and those for vitamin B3 were 13 and 15 mg/d, respectively. Among Japanese women aged 30–49 years, the corresponding estimated average requirement was 0·9 and 1·1 mg/d, and RDA was 10 and 12 mg/d. The estimated average requirement and RDA of vitamins B1 and B3 in Japanese men and women aged over 70 years were even less than any other age groups(Reference Shibata, Fukuwatari and Imai16). Not all individuals prefer or can afford vitamin supplements for their health; thus, improving the dietary vitamins intakes is more achievable and acceptable by the general population. Therefore, after established effects of supplementary intakes of vitamins B1 and B3 have been determined, studying the cardiovascular impacts of dietary intakes of vitamins B1 and B3 is now warranted. Owing to the research gap in the field of epidemiology and similar food sources of vitamins B1 and B3, their associations with CVD mortality were hypothesised in the present study. Therefore, we aimed to investigate the associations of dietary vitamins B1 and B3 intakes with risk of CVD mortality, which was considered as a proxy of CVD incidence risk, among Japanese men and women using the Japan Collaborative Cohort (JACC) study, a nationwide, community-based prospective cohort study.
Methods
Study population and baseline data
Under the sponsorship of the Ministry of Education, Sports, and Science, the JACC study had the baseline survey (1988–1990) of 110 585 Japanese men (n 46 395) and women (n 64 190) aged 40–79 years from forty-five areas all over Japan. A detailed cohort profile of the JACC study was published previously(Reference Tamakoshi, Ozasa and Fujino17). Data on the baseline lifestyle and participants’ characteristics, including demographic data, medical history of chronic diseases, diabetes mellitus, hypertension, smoking, alcohol consumption, exercise, diet and other items, were compiled via a self-administered questionnaire (online Supplementary Methods). The questionnaire included a validated forty-food item/FFQ which was distributed in thirty-two areas; therefore, we started with 86 401 subjects from those thirty-two areas. After the exclusion of non-respondents to FFQ (n 24 614), we further excluded those who reported a medical history of CVD or cancer (n 3142) and those who had implausible energy intakes defined as outliers of mean ± three standard deviations (n 343). Finally, a total of 58 302 individuals were eligible for the present study (22 989 men and 35 313 women) (online Supplementary Fig. S1). Written informed consent was acquired from community leaders or the individuals. The protocol of JACC study was approved by the Medical Ethical Committees of Nagoya University School of Medicine.
Dietary intake assessment
The participants were required to choose one from five frequency responses to describe the usual consumption frequency of forty food and beverage items over the past 12 months without specification of the portion size. The five responses were rarely, 1–2 times/month, 1–2 times/week, 3–4 times/week and almost every day. These frequencies were transformed into weekly consumption scores of 0, 0·38, 1·5, 3·5 and 7·0 per week, respectively(Reference Tamakoshi, Ozasa and Fujino17,Reference Date, Fukui and Yamamoto18) . A validation study among eighty-five individuals using four 3-d weighed dietary records over a 1-year period as a reference standard determined the portion size for each food and validated the FFQ intakes. The amount of nutrients in each food was calculated by multiplying the weekly consumption scores by the estimated portion size. The values of vitamins B1 and B3 and other nutrients from each food category were calculated according to the Standardised Tables of Food Composition, 5th revised version(Reference Hirokadzu19) which listed the nutrients content in 100 mg of different foods. Thus, the total vitamins B1 and B3 intakes were calculated by summing their intakes from all over the foods in the FFQ. The details of computation of nutrient intakes from FFQ(Reference Date, Fukui and Yamamoto18) and the accuracy of food composition tables in Japan(Reference Watanabe, Suzuki and Hagiwara20,Reference Hernando and Maria21) were published previously. The Spearman rank correlation coefficients for vitamins B1 and B3 intakes between the FFQ and the four 3-d dietary records were 0·36 and 0·32, respectively, after energy adjustment(Reference Eshak, Iso and Muraki22). The energy-adjusted mean ± standard deviation intakes in mg/d from weighed dietary record and FFQ were 1·08 (sd 0·20) and 0·71 (sd 0·20) for vitamin B1, but the respective values for vitamin B3 were not reported(Reference Date, Fukui and Yamamoto18).
Mortality surveillance
The investigators annually or biannually confirmed the dates and causes of death in each area(Reference Tamakoshi, Ozasa and Fujino17). The International Classification of Diseases, 10th revision (ICD10) codes were applied to determine the underlying causes of death. In this study, our primary outcome was the total CVD mortality (ICD I01-I99). Cause-specific outcomes included mortalities from total stroke (ICD I60-I69), haemorrhagic stroke (ICD I60-I61), ischaemic stroke (ICD I63.0-I63.9), ischaemic heart disease (ICD I20-I25), myocardial infarction (ICD I20) and heart failure (ICD I50). This death certificate ascertainment was applied to all deaths within our cohort except for deaths that occurred outside of the original resident areas, which were treated as censored cases.
Statistical analysis
Energy-adjusted dietary intakes of vitamins B1 and B3 were categorised into five categorical groups (quintiles). The significance of differences in means or proportions of participants’ characteristics and known risk factors of CVD in each quintile was tested by the ANCOVA and χ 2 test.
Person-years of follow-up were calculated from the baseline in 1988–1990 to their first endpoint in this follow-up as follows: death, moving out or the end of follow-up, whichever came first. The follow-up for mortality from CVD was conducted until 31 December 2009 in general; however, in four areas the follow-up was stopped until 31 December 1999, in another four areas until 31 December 2003 and in two areas until 31 December 2008(Reference Tamakoshi, Ozasa and Fujino17). The Cox proportional hazard model was applied to calculate crude and multivariable-adjusted hazard ratios and 95 % CI for risk of mortality from CVD during the follow-up period (1988–2009) across quintiles of dietary vitamins B1 and B3 intakes. We confirmed no violation of the Cox proportional hazard assumption because there were no significant interactions between the categorical rank variables of dietary vitamins B1 and B3 intakes and a function of survival time for all the tested outcomes. Multiplicative interactions of vitamins B1 and B3 with sex were tested to decide on presenting the data sex specifically or for combined men and women. The hypothesised confounders included age, sex, medical history of hypertension and diabetes, smoking status, ethanol intake, hours of sports, hours of walking, quintiles of BMI, years of education, perceived mental stress, daily utilisation of multivitamin supplementation, energy-adjusted quintiles of Na and SFA intakes and quintiles of total energy intake. Details of these factors are given in online Supplementary Methods.
We assigned the median values to each quintile of vitamins B1 and B3 and tested their significance to calculate the trends across quintiles of vitamins B1 and B3 intakes. We further conducted a sensitivity analysis by excluding those who died within first 3 years of follow-up to avoid potential as-yet-undiagnosed diseases at baseline. All probability values for statistical test were two-tailed, and P < 0·05 was regarded as statistically significant. We applied the SAS statistical package (Version 9.4; SAS Institute Inc.) for statistical analysis.
Results
As shown in Table 1, participants in the highest quintile of both vitamins B1 and B3 intake were older, less educated, under less mental stress, had more walking time, had higher BMI and were less likely to be current smoker and to have a history of hypertension or diabetes. They also used multivitamin supplementation less frequently and consumed less alcohol but consumed more Na, SFA and total energy when compared with those in the lowest quintile. In this study, sources of vitamin B1 were 31 % from pork, 17 % from vegetables, 10 % from fish and 7 % from potatoes, while sources of vitamin B3 were 43 % from fish, 13 % from vegetables, 8 % from pork, 7 % from coffee and 6 % from green tea (data not shown in tables).
Table 1. Participants’ characteristics and dietary variables according to quintiles of dietary vitamins B1 and B3 intakes at baseline in a cohort of 22 989 men and 35 313 women with a total of 3371 CVD mortality cases
Since no interaction with sex was observed for the association of vitamins B1 and B3 with CVD and specific endpoints, we combined the results of men and women in the main analyses. During 960 225 person-years of follow-up for 58 302 participants, we documented a total of 3371 deaths due to CVD, among whom there were 1504 deaths due to stroke (549 of which were due to haemorrhagic stroke and 816 of which were due to ischaemic stroke), 699 deaths were due to ischaemic heart disease (including 524 deaths due to myocardial infarction) and 564 deaths were due to heart failure.
As shown in Table 2, the dietary intake of vitamin B1 was not associated with mortality from total stroke or its subtypes. On the other hand, a higher dietary vitamin B1 intake was associated with the reduced risk of ischaemic heart disease, myocardial infarction and total CVD; hazard ratios were 0·57 (95 % CI 0·40, 0·80; P for trend < 0·01), 0·56 (95 % CI 0·37, 0·82; P for trend < 0·01) and 0·85 (95 % CI 0·73, 0·99; P for trend = 0·03), respectively, in the highest v. lowest intake quintile. Moreover, the multivariable-adjusted hazard ratio of heart failure mortality in the highest v. lowest intake quintiles was 0·65 (95 % CI 0·45, 0·96; P for trend = 0·13).
Table 2. CVD mortality according to quintiles of vitamin B1 intake (Hazard ratios and 95 % confidence intervals)
1Adjusted for age, sex, and socio-economic status (educational status).
2Adjusted for age, sex, socio-economic status (educational status), and health behaviours (hours of walking, hours of sports, ethanol intake and smoking status).
3A full mode with adjustment for age, sex, educational status, hours of walking, hours of sports, ethanol intake, smoking status, history of hypertension and diabetes, BMI, perceived mental stress, multivitamin supplementation, quintiles of energy-adjusted Na and SFA intakes and total energy intakes.
For vitamin B3, as shown in Table 3, there was no association with the mortality from stroke or heart failure. Statistically significant inverse trends in risks of mortality from total CVD, haemorrhagic stroke, ischaemic heart disease and myocardial infarction were observed in the age- and sex-adjusted model. However, after the multivariate adjustment, these associations were weakened; the multivariable-adjusted hazard ratios in the highest v. lowest quintiles of dietary vitamin B3 were 0·90 (95 % CI 0·80, 1·03; P for trend = 0·13) for total CVD mortality, 0·74 (95 % CI 0·55, 1·01; P for trend = 0·16) for haemorrhagic stroke, 0·79 (95 % CI 0·60, 1·04; P for trend = 0·05) for ischaemic heart disease and 0·66 (95 % CI 0·48, 0·90; P for trend = 0·02) for myocardial infarction.
Table 3. CVD mortality according to quintiles of vitamin B3 intake (Hazard ratios and 95 % confidence intervals)
1Adjusted for age, sex and socio-economic status (educational status).
2Adjusted for age, sex, socio-economic status (educational status) and health behaviours (hours of walking, hours of sports, ethanol intake and smoking status).
3A full mode with adjustment for age, sex, educational status, hours of walking, hours of sports, ethanol intake, smoking status, history of hypertension and diabetes, smoking status, BMI, hours of walking, hours of sports, educational status, perceived mental stress, ethanol intake, multivitamin supplementation, quintiles of energy-adjusted Na and SFA intakes and total energy intakes.
There were 456 participants who died within the first 3 years of follow-up, and excluding those subjects yielded no substantial changes in the associations of vitamins B1 and B3 with mortality from ischaemic heart disease and myocardial infarction (online Supplementary Table S1).
Discussion
In this large community-based prospective cohort study of Japanese men and women, higher dietary intakes of vitamins B1 and B3 were associated with reduced risks of mortality from total CVD, ischaemic heart disease and myocardial infarction. Neither dietary vitamin B1 nor vitamin B3 intake was associated with the mortality risk of stroke, except for a tendency towards a reduced risk of haemorrhagic stroke with a higher dietary vitamin B3 intake. Moreover, a higher dietary intake of vitamin B1 was associated with a reduced risk of heart failure.
As far to our knowledge, the present study is the first to investigate associations of dietary vitamins B1 and B3 intakes with risk of CVD mortality despite the abundant evidence from animal studies and clinical trials on vitamins B1 and B3 supplements. Vitamins B1 and B3 in animal studies and human clinical trials showed protective effects against myocardial ischaemia.
One study on dogs showed that administration of vitamin B1 decreased the metabolic needs of the heart, which was manifested as reduced myocardial oxygen consumption, mean peripheral pressure and left ventricular pressure up to 45, 25 and 10 % respectively(Reference Freye11). A clinical trial on ten healthy adults and ten type 2 diabetes patients reported improvements in the brachial artery vasoactivity and the endothelium-dependent vasodilatation in both groups after a week of daily intravenous administration of 100 mg of vitamin B1 (Reference Arora, Lidor and Abularrage23). Another randomised, cross-over and investigator-blinded trial on twenty adult healthy volunteers indicated the flow-mediated dilatation of the brachial artery was reduced by 50 % of its baseline diameter after smoking one cigarette, and the reduction in the flow-mediated vasodilatation with smoking one cigarette was only 25 % when 1050 mg/d oral benfotiamine was administered for 3 d before the experiment(Reference Stirban, Nandrean and Kirana24).
On the other hand, vitamin B3 is a candidate to lower the risk of CVD as it is known to decrease LDL-cholesterol, TAG and lipoprotein(a) levels, while increasing HDL-cholesterol level(Reference McKenney7). Among 8341 American men aged 30–64 years from Coronary Drug Project with previous myocardial infarction, 3000 mg/d vitamin B3 v. placebo for a follow-up of 15 years reduced 14 % of the mortality from total CVD and 26 % of the mortality from ischaemic attack after a mean follow-up of 15 years(Reference Canner, Berge and Wenger12). Additionally, a meta-analysis of twenty-three randomised controlled trials including 39 195 participants reported a pooled risk ratio (CI) of mortality from fatal or non-fatal myocardial infarction (OR: 0·93; 95 % CI 0·87, 1·00) for vitamin B3 (median dose: 2 g/d; median duration: 11·5 months) v. control(Reference Schandelmaier, Briel and Saccilotto13). Another meta-analysis of eleven randomised controlled trials including 6616 participants showed vitamin B3 (250–3000 mg/d) significantly decreased major coronary events (OR: 0·75; 95 % CI 0·65, 0·96), stroke (OR: 0·74; 95 % CI 0·59, 0·92) and any cardiovascular events (OR: 0·73; 95 % CI 0·63, 0·85)(Reference Bruckert, Labreuche and Amarenco9). In a recent meta-analysis of seventeen clinical trials including 35 760 participants, vitamin B3 therapy (100–4000 mg/d) was shown to be associated with reduction of acute coronary syndrome (relative risk: 0·74; 95 % CI 0·58, 0·96) and stroke (relative risk: 0·74; 95 % CI 0·59, 0·94)(Reference D’Andrea, Hey and Ramirez25). A meta-analysis including 9959 subjects reported similar results for total CVD events (OR: 0·66; 95 % CI 0·49, 0·89) and major coronary events (OR: 0·75; 95 % CI 0·59, 0·96) but not for stroke (OR: 0·88; 95 % CI 0·50, 1·54)(Reference Lavigne and Karas26). Another meta-analysis of thirteen trials of vitamin B3 treatment demonstrated a significant reduced risk of non-fatal myocardial infarction (risk ratio: 0·85; 95 % CI 0·73, 1·00), a weak association with CVD mortality (risk ratio: 0·91; 95 % CI 0·81, 1·02) and no association with stroke (risk ratio: 0·89; 95 % CI: 0·72, 1·10)(Reference Garg, Sharma and Krishnamoorthy27).
The mechanisms by which vitamins B1 and B3 might be protective against CVD mortality, especially those from ischaemic heart disease could be summarised here. Vitamin B1 deficiency was highly prevalent in patients with type 2 diabetes(Reference Thornalley, Babaei-Jadidi and Al Ali28), which is considered as one of the risk factors for ischaemic heart disease. In addition, vitamin B1 inhibits human infragenicular accelerated proliferation of arterial smooth muscle cells and mitigates atherosclerosis and endothelial dysfunction(Reference Avena, Arora and Carmody29). Another potential mechanism might be the protective effects of vitamin B1 against ischaemic injury via reducing the metabolic needs of heart(Reference Yamada, Kusakari and Akaoka5). For vitamin B3, the reduced CVD risk may be involved in the favourable effects of vitamin B3 on lipid metabolism(Reference McKenney7,Reference Linder, Willmann and Kantartzis8) . Vitamin B3 also has anti-inflammatory properties demonstrated by lowering C-reactive protein lipoprotein-associated phospholipase A2, inhibiting pro-atherogenic chemokines and enhancing serum levels of adiponectin(Reference Lavigne and Karas26). Moreover, an antihypertensive effect of vitamin B3 was also suggested(Reference Bays, Maccubbin and Meehan30).
We observed that a higher dietary vitamin B1 intake was associated with reduced risk of mortality from heart failure. Vitamin B1 deficiency was commonly considered to be correlated with a failing heart. In a meta-analysis of nine observational studies, the prevalence of vitamin B1 deficiency was higher with an OR of 2·5 (95 % CI 1·7, 3·9) in heart failure group than in control(Reference Jain, Mehta and Al-Ani31). Also known as wet beriberi or cardiac beriberi, vitamin B1 deficiency was characterised by peripheral neuropathy and muscle weakness resulting in heart failure(Reference Eshak and Arafa3). The vitamin B1 deficiency-related heart failure was attributed to the vitamin B1 role in energy metabolism(Reference Eshak and Arafa3). Some studies reported that vitamin B1 supplementation had beneficial effects on cardiac function(Reference Schoenenberger, Schoenenberger-Berzins and der Maur6), but the evidence is still inconclusive(Reference Eshak and Arafa3).
The dietary intake of vitamin B3 in our study tended to associate with a lower risk of mortality from haemorrhagic stroke. In a case–control study including sixty-nine stroke cases and sixty-nine controls, vitamin B3 was found to be inversely correlated with risk of stroke (OR: 0·17; 95 % CI 0·04, 0·82)(Reference Park32). However, some meta-analyses concluded that vitamin B3 had similar protective effects on both CHD and stroke outcomes(Reference Bruckert, Labreuche and Amarenco9,Reference D’Andrea, Hey and Ramirez25) , while others failed to find protective effects against stroke(Reference Lavigne and Karas26,Reference Garg, Sharma and Krishnamoorthy27) . The available studies and meta-analyses did not comment on the effect of vitamin B3 on stroke subtypes. Vitamin B3 was shown to reduce the blood pressure, an important risk factor for haemorrhagic stroke(Reference Bays, Maccubbin and Meehan30). On the other hand, vitamin B3 may promote vascular plasticity after an acute attack of stroke. In an animal experiment, Niaspan treatment of stroke in rats with diabetes promoted vascular remodelling and improved functional outcome(Reference Ye, Chopp and Cui33). Therefore, an impact of vitamin B3 on risk of haemorrhagic stroke mortality could be plausible.
To the best of our knowledge, this is the first study to investigate the association of dietary intakes of vitamins B1 and B3 with the risk of mortality from CVD among Japanese population. The JACC study is a large, nationwide, community-based, prospective Japanese cohort study. The large sample size allowed us to investigate the associations of quintile categories of dietary vitamins B1 and B3 intakes with the risks of type-specific cardiovascular mortality as well as total CVD in Japanese population. Other strengths of this study included the prospective study design, the utilisation of a validated FFQ, the consistent endpoint determination and the exclusion of participants with CVD and cancer before the starting point of follow-up.
Limitations of this study mainly originate from the dietary assessment. The one-time measurement of dietary intakes cannot completely represent the consumption of nutrients during a long-term follow-up. The exclusion of 18 428 non-respondents to FFQ might result in a selection bias. Compared with 24 614 non-respondents to FFQ, the 61 787 respondents were more likely to be young and more educated (online Supplementary Table S2). Several research of the JACC study reported the underestimation of nutrients intakes, which could be attributed to the limited number of food items in the used FFQ. Second, we had no data about the exact amounts or types of vitamin supplementation. To our knowledge, in the past century, vitamin supplementation was not common among Japanese population; thus, we believe that it would not affect the result substantially. In this study, approximately 88 % of participants did not use any vitamin supplementation and only 3 % used it on daily basis. The exclusion of daily supplementation uses did not change the result. Third, in Japan, the accuracy of heart failure death certificate diagnosis is a questionable issue. It is generally believed that heart failure death was overestimated before 1994, because most deaths of unknown origin such as cardiac arrest or arrhythmic death were more likely diagnosed as unspecific heart failure(Reference Eshak, Iso and Yamagashi34). Therefore, approximately 27–50 % heart failure deaths were accounted for this misclassification(Reference Eshak, Iso and Yamagashi34). Lastly, we did not have data on biomarkers of atherosclerosis and endothelial dysfunction, lipid metabolism and systematic inflammation such as C-reactive protein, lipoprotein-associated phospholipase A2, pro-atherogenic chemokines or serum levels of adiponectin and cannot determine all confounding effects from some other nutrients, lifestyles and socio-economic factors.
Conclusions
In this prospective cohort study, higher dietary intakes of vitamins B1 and B3 were inversely associated with a reduced risk of mortality from CVD among Japanese men and women. Dietary intakes of these vitamins from their food sources are available, accessible, affordable, safe and more acceptable by the general population than supplementary intakes. Therefore, dietary intakes of food rich in these vitamins could be encouraged for decreasing the risk of CVD mortality. However, our findings warrant further studies in different populations.
Acknowledgements
The authors thank Dr. Kunio Aoki, Professor Emeritus, Nagoya, University School of Medicine and the former Chairman of the JACC Study, and Dr. Haruo Sugano, the former Director of the Cancer Institute, Tokyo, who greatly contributed to the initiation of the JACC study. The authors also thank all staff members involved in this study for their valuable help in conducting the baseline survey and follow-up.
This study has been supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT) (MonbuKagaku-sho); Grants-in-Aid for Scientific Research on Priority Areas of Cancer and Grants-in-Aid for Scientific Research on Priority Areas of Cancer Epidemiology from MEXT (nos. 61010076, 62010074, 63010074, 1010068, 2151065, 3151064, 4151063, 5151069, 6279102, 11181101, 17015022, 18014011, 20014026, 20390156, 26293138), and JSPS KAKENHI No.16H06277. This research was also supported by Grant-in-Aid from the Ministry of Health, Labour and Welfare, Health and Labor Sciences research grants, Japan (Comprehensive Research on Cardiovascular Disease and Life-Style Related Diseases: H20–Junkankitou (Seishuu)–Ippan–013; H23–Junkankitou (Seishuu)–Ippan–005); an Intramural Research Fund (22-4-5) for Cardiovascular Diseases of National Cerebral and Cardiovascular Center; Comprehensive Research on Cardiovascular Diseases and Life-Style Related Diseases (H26-Junkankitou (Seisaku)-Ippan-001) and H29–Junkankitou (Seishuu)–Ippan–003 and 20FA1002.
C. T. designed the study, analysed the data and wrote the manuscript. E. S. E. and H. I. conducted the technique review and reviewed and edited the manuscript. K. S. and A. T. made critical revisions of the manuscript. C. T. and H. I. had primary responsibility for final content. All authors read and approved the final manuscript.
The authors have no conflict of interest to declare.
Supplementary material
For supplementary material referred to in this article, please visit https://doi.org/10.1017/S0007114522001209
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