Soy and fish, first focused experimentally on their preventive effect on stroke, were epidemiologically investigated worldwide for the association of the biomarkers of their intakes, isoflavones and taurine, in 24-h urine (24U) with cardiovascular disease risks. Since these biomarkers were inversely related with coronary heart disease mortality, cross-sectional and cohort studies, particularly among Japanese regularly taking soy and fish diets, proved that soy and fish intakes estimated by their biomarkers in 24U or dietary questionnaires were related with lower cardiovascular risks. Intervention studies of soy and/or fish indicated cardiovascular risk reduction and the effective mechanisms involved in it.

Objective: Since nutrition could extend lifespan even in stroke-prone rats developing stroke genetically, our WHO-Collaborating Center for Primary Prevention of Cardio-vascular Diseases (CVD) started world-wide hearth examination to investigate nutrition good for healthy longevity by collecting 24-hour urine (24U) samples. Design: Urinary biomarkers of nutritions, sodium for salt, potassium (K) for vegetables and fruits, isoflavones(Is) for soy, taurine(Ta) for fish and magnesium (Mg) for beans and nuts were analyzed in 24U among 4211 participants (49.7% females(F)), aged 48–56 years in 50 populations from 22 countries according to WHO-CARDIAC(Cardiovascular Diseases and Alimentary Compression) Study Protocol, and studied comparatively in 6 Japanese(J), 6 Mediterranean(M) populations from Greece, Italy(2), Spain(2), Portugal and 6 Euro-Western(EW) populations from Sweden, Ireland, Scotland, Canada, Australia, New Zealand. Results with Discussion: The quintile analyses of Ta and Mg/creatine ratio in the world were inversely associated with CVD risks. The Odds ratios of obesity, hypercholesterolemia and hypertension in the lowest quintile of both Ta and Mg were 6.3, 4.6 and 1.8, respectively. J and M populations had significantly higher Ta and Mg therefore, lower CVD risks than EW, but J and M took more salt increasing the risk of stroke (Table 1). Despite the high salt intake, M had the merit of significantly high K intake to attenuate the adverse salt effect. J had a merit to take Is from soy containing Mg. Since Is intake was inversely associated with lower BMI in the world populations, high Japanese Mg intake was related to high Is intake which was inversely associated with obesity in Japan. In fact, the Japanese populations who took significantly higher T and Mg had lower BMI as well as higher HDL and folic acid in the blood, expected to prevent atherosclerosis and dementia. In contrast to M and EW, particularly severe obesity over BMI 30 was obviously less in J, ranking 182nd among 190 populations (WHO). The mortality rates of COVID-19 were significantly positively related to the ratio of severe obesity among 20 major countries in the world and this mortality was obviously lower in J, suggestive of the potential of J diet for post-pandemic longevity. Conclusion: 24U biomarker analyses indicated low-salted J and M diets are recommendable for CVD prevention. J diet is so far better than M diet because of lower CVD risks such as obesity and lipidemia, which were proven by urinary biomarkers to be related with traditional fish and soy intakes.

Budgerigar fledgling disease virus (BFDV) represents the first avian member of the Polyomavirus family. In contrast to mammalian polyomaviruses BFDV exhibits unique biological properties, in particular it is able to cause an acute disease with distinct organ manifestations in affected birds. Here we present the complete nucleotide sequence of the BFDV genome, consisting of 4980 bp. When compared to published nucleotide sequences of other polyomaviruses, the BFDV genome exposes a number of very similar structural features, and undoubtedly qualifies as a member of that family of viruses. The most important differences include a large T antigen remarkably reduced in size, and an origin of replication region with fundamental deviations from the origin structure of all other polyomaviruses. The specific characteristics of the BFDV genome may be used to place this virus into a distinct subgroup within the Polyomavirus family and may give a clue to the elucidation of its extraordinary biological properties.

To investigate the association of the Japanese diet with risks for lifestyle-related diseases, the biomarkers of seafood and soybean consumption, taurine (T) and soy isoflavones (I), and others were analyzed in 24-hour urine (24U) samples collected from participants of the Cardiovascular Diseases and Alimentary Comparison (CARDIAC) Study coordinated by the World Health Organization (WHO). The data of T and I normalized for creatinine content in 24U were divided into five quintiles, T1 to T5, and I1 to I5. The total data of the collected samples were divided into 25 groups, which were obtained by 5 (T1-T5) × 5 (I1-I5) according to 24U excretions of T and I corresponding to the intake of seafood and soybeans from the least to the highest, respectively. Since these two nutrients were often consumed together in the Japanese diet, this characteristic was expressed as J1 to J5 based on the amounts of 24U T and I excretions. The risks for lifestyle-related diseases, obesity (body mass index, BMI), and cholesterolemia became lower during the transition from J1 to J5, while HDL cholesterol levels became higher from J1 to J5. On the contrary, urinary salt excretion and the sodium (Na)/potassium (K) ratio became higher from J1 to J5. Systolic blood measure was significantly lower in J3 than in J5. Diastolic blood pressure was also significantly lower in J3 than in J1. In conclusion, the higher the J score, which corresponds to Japanese dietary habits, the lower the BMI and cholesterol levels, as well as mortality rate from coronary heart disease, but the higher the average life expectancy among the Japanese. However, these higher J scorings were associated with high-salt intake and high Na/K ratios; therefore, they contributed to high blood pressure and high mortality rate caused by stroke in Japan. These results indicate that low-salt intake should be recommended to the Japanese who are consuming seafood and soy regularly in order to maintain lower blood pressure and to extend healthy life expectancy with a lower risk of stroke. Moreover, high scorings of the Japanese diet correspond to the high intake of magnesium (Mg) which is rich in seafood including seaweeds and soy. Therefore, low-salt seafood and soy intake is expected to reduce the incidence of the metabolic syndrome, the risk of which is inversely related to T and Mg intake.

Plant polyphenols have various health effects. Genistein, which is abundant in soybeans, and epigallocatechin-3-gallate, which is abundant in green tea, are major flavonoids, a subclass group of polyphenols. Several epidemiological studies have shown that these flavonoids have beneficial effects against cancer and cardiovascular diseases. However, other studies did not show such effects. Several confounding factors, including recall bias, are related to these inconsistent findings, and the determination of metabolites in the urine may be useful in reducing the number of confounding factors. Equipment, which can be used by research participants to collect samples from a portion of voided urine within 24 h without the help of medical workers, has been developed for epidemiological investigations. Previous studies, in which flavonoid metabolites in these urine samples were measured, revealed that soy intake was correlated with a reduced risk of certain types of cancer and cardiovascular diseases worldwide. Although soybeans and green tea consumption may have protective effects against cancer and cardiovascular diseases, further clinical studies that consider different confounding factors are required to provide evidence for the actual impact of dietary flavonoids on human diseases, including cancer and cardiovascular diseases. One possible mechanism involved is discussed in relation to the downregulation of reactive oxygen species and the upregulation of 5′-adenosine monophosphate-activated protein kinase elicited by these flavonoids.

Objective: Since stroke was prevented by taurine(T)- and magnesium(Mg)-rich diets even in salt-loaded stroke-prone SHR models, we examined the association of 24-hour urinary(24U) T and Mg/creatinine (C) ratios with cardiovascular risks (CVR) world-wide and studied these biomarkers in Japanese and Mediterranean populations (J, M) in comparison with Euro-Western populations(EW). Design: The association of 24U T and Mg/C with CVR was cross-sectionally analyzed among 4211 participants, 49.7% females(F), aged 48–56 years in 50 populations from 22 countries according to the WHO-CARDIAC study protocol and further analyzed comparatively in 6 J (864, 53.8%F), 6 M(574, 50.2%,F from Greece, Sicily, Italy, Spain, Portugal) and 6 EW(549,45.9% F from Sweden,Ireland, Scotland, Canada, Australia,New Zealand). Design and method: In a world-wide linier regression analysis, T/C and Mg/C were inversely associated with BMI, systolic and diastolic blood pressure (SBP, DBP) and serum total cholesterol (TC) significantly (P for linear trend <0.001, respectively). The prevalence of obesity, hypertension and hypercholesterolemia among the subjects of the lowest quintile of T/C (Mg/C) was 2.84 (2.49), 1.22 (1.49) and 2.20 (2.39) times higher (P < 0.05 or <0.001) than those in the subjects of the highest quintile. In a comparison of J and M (both males and females) with EW, 24U T and Mg/C in both populations were significantly higher than in EW, and SBP, DBP, TC and non-HDL cholesterol were significantly lower in J and M, despite their significantly higher 24U salt, than in EW, indicating that these biomarkers could contribute to lower coronary heart disease mortality, benefiting longevity in J and M. Conclusion: T and Mg are regarded as beneficial dietary factors for reducing CVR world-wide and are related with the merits of J and M diets compared with EW diets despite the higher salt intake, indicating that low salted T- and Mg-rich dietary intakes are recommendable for promoting CVR reduction.

Levels of isoflavones, biomarkers of soy intake, in 24-hour urine (24U) were inversely related to coronary heart disease (CHD) mortality in the World Health Organization's Cardiovascular Disease and Alimentary Comparison Study. Considering 24 U isoflavone levels were highest and CHD mortality was lowest in the Japanese, who maintained the world's longest life expectancy, the association of regular soy intake with cardiometabolic risk was investigated in Japanese adults (20-49 years old) and elderly (50-79 years old). In multivariate analysis adjusted for age, sex, and drug treatments, mean 24 U isoflavone excretion was significantly inversely associated with insulin resistance in the elderly and significantly associated with blood folate and potassium in the elderly, but also positively associated with 24 U salt in the elderly. These findings indicate that low-salt soy should be recommended to improve glucose metabolism in elderly Japanese.

In the World Health Organization (WHO)-coordinated Cardiovascular Disease and Alimentary Comparison Study, isoflavones (I; biomarker for dietary soy) and taurine (T; biomarker for dietary fish) in 24-hour-urine (24U) were inversely related to coronary heart disease (CHD) mortality. High levels of these biomarkers are found in Japanese people, whose CHD mortality is lowest among developed countries. We analyzed the association of these biomarkers with cardiovascular disease risk in the Japanese to know their health effects within one ethnic population. First, to compare the Japanese intake of I and T with international intakes, the ratios of 24UI and 24UT to creatinine from the WHO Study were divided into quintiles for analysis. The ratio for the Japanese was the highest in the highest quintiles for both I and T, reaching 88.1%, far higher than the average ratio for the Japanese (26.3%) in the total study population. Second, 553 inhabitants of Hyogo Prefecture, Japan, aged 30 to 79 years underwent 24-U collection and blood analyses. The 24UT and 24UI were divided into tertiles and adjusted for age and sex. The highest T tertile, compared with the lowest tertile, showed significantly higher levels of high-density lipoprotein-cholesterol (HDL-C), total cholesterol, 24U sodium (Na) and potassium (K). The highest I tertile showed significantly higher folate, 24UNa and 24UK compared with the lowest tertile. The highest tertile of both T and I showed significantly higher HDL-C, folate, and 24UNa and 24UK compared with the lowest tertile. Thus, greater consumption of fish and soy were significantly associated with higher HDL-C and folate levels, possibly a contributor to Japan having the lowest CHD mortality and longest life expectancy among developed countries. As these intakes were also associated with a high intake of salt, a low-salt intake of fish and soy should be recommended for healthy life expectancy.

WHO-CARDIAC (Cardiovascular Diseases and Alimentary Comparison) Study revealed the quintile analyses of 24-h urinary (24 U) taurine (T) and magnesium (Mg) excretions were inversely related with cardiometabolic risks (CMR) such as obesity, hypertension and hypercholesterolemia in 50 population samples in the world. To exclude the influence of ethnicity in the study, 24 U T and Mg excretions were analyzed for the association with CMR in one ethnicity, Japanese population. 24 U T/creatinine (C) ratios were divided into 5 quintiles and the ratios of Japanese to the total of each quintile were analyzed from CARDIAC Study samples. The highest 24 U T quintile consisted of 60% Japanese, indicating high seafood consumption in Japanese. Over 600 Japanese aged 30-79 were invited to a health examination for blood pressure measurement and for fasting blood and 24 U samplings. Tertile analysis of 24 U T/C ratios in relation to CMR indicated the third tertile had significantly higher HDL cholesterol, 24 U potassium (K) and 24 U salt than the first (lowest) tertile. Tertile analysis of 24 U Mg/C ratios indicated the third tertile had significantly lower body mass index and significantly higher folic acid, 24 U isoflavones, K and salt than the first tertile after age and gender adjustment. The third tertile of both T/C and Mg/C had significantly lower body mass index, LDL/HDL and Na/K ratios, and significantly higher HDL cholesterol and folic acid than the first tertile, - indicating seafood eaters taking Mg rich diets had lower risks of obesity, atherosclerosis, hypertension and higher folic acid, beneficial for healthy longevity.

[This corrects the article DOI: 10.1371/journal.pone.0176039.].

Detection of individual difference in salt-induced blood pressure (BP) elevation is important to know who should reduce salt intake for preventing hypertension, but the diagnosis of salt sensitivity requires a time-consuming protocol for testing BP changes in response to salt loading and depletion. Since WHO-CARDIAC Study indicated BP was higher in the people excreting more 24-hour urinary (24U) salt with higher heart rate (HR), ambulatory BP was monitored in a Korean population to investigate the association of nighttime BP and HR with 24-hour urinary salt excretion for detecting salt sensitivity.Total 101 males and females aged 30 to 61 years were examined for ambulatory BP (by TM2430, A&D, Tokyo, Japan) and 24U collected by aliquot cups.In the participants excreting 24U salt equal to or higher than (≥) the mean, 10.8 g, nighttime systolic and diastolic BP (SBP, DBP), in the group with HR ≥ the mean, 64.7 beats/min (18) were 119.8/77.1 mmHg, significantly higher than 109.2/67.8 mmHg in the group with HR.Significant nighttime BP elevation could be detected by 24U salt ≥ 10 g and nighttime HR ≥ 65 in this Korean population. Collecting 24U by using a simple aliquot cup and monitoring nighttime HR by a wrist watch-type HR detector are expected to detect salt sensitivity also in other populations, as noted in significantly higher BP in 50 CARDIAC Study populations with higher 24U salt and HR than their means.

OBJECTIVE OBJECTIVE It is now established that "dippers" whose blood pressure (BP) falls at night have lower cardiovascular risks than nondippers without nocturnal BP fall. Our previous study on the middle-aged general population in Korea indicated increased 24-hour urinary (24U) sodium excretion was associated with increased nighttime BPs. Therefore, we further analyzed the association of salt intakes estimated by 24U analysis with dipping in nocturnal BP and heart rate. DESIGN AND METHOD DESIGN AND METHOD In the rural area in South Korea 218 subjects aged 30 to 59 years were measured with casual and ambulatory BP monitoring (ABPM) (TM2430, A&D, Tokyo, Japan) and their 24U samples collected by aliquot cups were analyzed. RESULTS For both ABPM and 24U analyses, 148 subjects, aged 47.4 ± 8.3 years in average and female rates 57.4%, were accepted for data analysis by the 24U creatinine equation. Salt intake, previously proven to be associated with nighttime systolic and diastolic BP but not with casual and daytime BP, was further analyzed to be significantly inversely associated not only with nocturnal dipping in systolic and diastolic BP (P < 0.01, 0.001), but also in heart rate (P < 0.01). Correlations of the dipping of systolic (Fig) and diastolic BPs with heart rate were significant (P < 0.001, 0.01). CONCLUSIONS CONCLUSION Higher salt intake attenuates dipping not only in BP but also in heart rate. Nocturnal heart rate monitoring, more easily available than ABPM, may be useful for detecting nocturnal BP rise and salt sensitivity related to cardiovascular risks.

OBJECTIVEDetection of individual difference in salt-induced blood pressure (BP) elevation is important to know who should reduce salt intake for preventing hypertension, but the diagnosis of salt sensitivity requires a time-consuming protocol for testing BP changes in response to salt loading and depletion. Since WHO-CARDIAC Study indicated BP was higher in the people excreting more 24-hour urinary (24U) salt with higher heart rate (HR), ambulatory BP was monitored in a Korean population to investigate the association of nighttime BP and HR with 24-hour urinary salt excretion for detecting salt sensitivity.DESIGN AND METHODTotal 101 males and females aged 30 to 61 years were examined for ambulatory BP (by TM2430, A&D, Tokyo, Japan) and 24U collected by aliquot cups.RESULTSIn the participants excreting 24U salt equal to or higher than (≥) the mean, 10.8 g, nighttime systolic and diastolic BP (SBP, DBP), in the group with HR ≥ the mean, 64.7 beats/min (18) were 119.8/77.1 mmHg, significantly higher than 109.2/67.8 mmHg in the group with HR.CONCLUSIONSSignificant nighttime BP elevation could be detected by 24U salt ≥ 10 g and nighttime HR ≥ 65 in this Korean population. Collecting 24U by using a simple aliquot cup and monitoring nighttime HR by a wrist watch-type HR detector are expected to detect salt sensitivity also in other populations, as noted in significantly higher BP in 50 CARDIAC Study populations with higher 24U salt and HR than their means.

We previously showed that 24-h urinary taurine (Tau) excretion was inversely associated with mortality due to coronary heart diseases (CHD) and stroke. The aim of this study was to examine the association between 24-h urinary Tau/creatinine (Cre) ratio and cardiovascular disease risk factors, including body mass index (BMI), blood pressure (BP), serum total cholesterol (TC) and prevalence of obesity, hypertension and hypercholesterolemia.

Serum, plasma and dietary magnesium (Mg) have been reported to be inversely associated with cardiovascular disease risk factors. We examined the associations between the 24-h urinary Mg/creatinine (Cre) ratio and cardiovascular disease risk factors, such as body mass index (BMI), blood pressure (BP), serum total cholesterol (TC) and prevalence of obesity, hypertension and hypercholesterolemia. A cross-sectional analysis was conducted among 4211 participants (49.7% women) aged 48-56 years in 50 population samples from 22 countries in the World Health Organization-coordinated Cardiovascular Diseases and Alimentary Comparison (CARDIAC) Study (1985-1994). In linear regression analyses, Mg/Cre ratio was inversely associated with BMI, systolic BP (SBP), diastolic BP (DBP) and TC (P for linear trend <0.001 for each). These associations were not markedly altered by adjustment for traditional risk factors, urinary markers or cohort effects. Multivariate-adjusted mean values for the subjects in the highest Mg/Cre ratio quintile were 6.3, 3.4, 5.3 and 4.6% lower than those for the subjects in the lowest quintile for BMI, SBP, DBP and TC (P<0.001, respectively). The prevalence of obesity, hypertension and hypercholesterolemia was 2.10 (95% confidence interval: 1.50, 2.95), 1.55 (1.25, 1.92) and 2.06 (1.63, 2.62) times higher (P<0.001, respectively) among the subjects in the lowest Mg/Cre ratio quintile than in the subjects in the highest quintile. These associations were not appreciably altered by adjustment for potential confounding variables. In conclusion, higher 24-h urinary Mg/Cre ratio was associated with lower cardiovascular disease risk factors, including BMI, BP, TC, obesity, hypertension and hypercholesterolemia.

To investigate the effects of daily supplementation with docosahexaenoic acid (DHA) on coronary heart disease risks in 38 middle-aged men with hypertension and/or hypercholesterolemia in Scotland, a five-week double-blind placebo-controlled dietary supplementation with either 2 g of DHA or active placebo (1 g of olive oil) was conducted. Percent composition of DHA in plasma phospholipids increased significantly in DHA group. Systolic and diastolic blood pressure and heart rate decreased significantly in DHA group, but not in placebo group. High-density lipoprotein cholesterol (HDL-C) increased significantly, and total cholesterol (TC)/HDL-C and non-HDL-C/HDL-C ratios decreased significantly in both groups. There was no change in TC and non-HDLC. We conclude that 2 g/day of DHA supplementation reduced coronary heart disease risk factor level improving blood pressure, heart rate, and lipid profiles in hypertensive, hypercholesterolemic Scottish men who do not eat fish on a regular basis.

The initial observation that taurine (T) prevented stroke in stroke-prone spontaneously hypertensive rats (SHRSP) led us to study the effects of T on cardiovascular diseases (CVD), as well as the epidemiological association of T and mortality rates, by using the data from WHO-coordinated Cardiovascular Disease and Alimentary Comparison Study, which covered 61 populations in 25 countries. In this study, 24 hour urine (24-U) samples were examined along with biomarkers of CVD risk. The mortality rate from ischemic heart disease (IHD), which was lowest among the Japanese compared to the populations of other developed countries, was positively related to total serum cholesterol (TC) and inversely related to 24-U taurine excretion (24-UT), as well as the n-3 fatty acid to total phospholipids ratio of the plasma membrane, both biomarkers of seafood intake. Analysis of 5 diet-related factors revealed that TC and BMI were positively associated with IHD mortality in both genders while Mg and T were negatively associated with IHD mortality. TC and sodium (Na) were negatively and positively associated with stroke mortality, respectively. 24-UT was negatively associated with stroke mortality. These five diet-related factors explained 61 and 49% of IHD and stroke variances in male, 63 and 36% of IHD and stroke variances in female, respectively.