Fruit, vegetable and bean intake and mortality from cardiovascular disease among Japanese men and women: the JACC Study.

To examine the association of plant-based food intakes with CVD and total mortality among Japanese. In the Japan Collaborative Cohort Study for Evaluation of Cancer Risk, 25 206 men and 34 279 women aged 40-79 years, whose fruit, vegetable and bean intakes were assessed by questionnaire at baseline in 1988-90, were followed for 13 years. Deaths from total stroke, stroke subtypes, CHD and total CVD, according to the International Classification for Diseases 10th Revision, were registered. During 756 054 person-years of follow-up, there were 559 deaths from total stroke, 258 from CHD, 1207 from total CVD and 4514 from total mortality for men, and for women, 494, 194, 1036 and 3092, respectively. Fruit intake was inversely associated with mortality from total stroke (the multivariable hazard ratio (HR (95 % CI)) in the highest v. lowest quartiles = 0.67 (0.55, 0.81)), total CVD (HR = 0.75 (0.66, 0.85)) and total mortality (HR = 0.86 (0.80, 0.92)). Vegetable intake was inversely associated with total CVD (HR = 0.88 (0.78, 0.99)). Bean intake was inversely associated with other CVD (HR = 0.79 (0.64, 0.98)), total CVD (HR = 0.84 (0.74, 0.95)) and total mortality (HR = 0.90 (0.84, 0.96)). Further adjustment for other plant-based foods did not alter the association of fruit intake with mortality from total stroke, total CVD and total mortality, but attenuated the associations of vegetables and beans with mortality risk. In conclusion, intakes of plant-based foods, particularly fruit intake, were associated with reduced mortality from CVD and all causes among Japanese men and women.

Protein tyrosine kinase 2beta as a candidate gene for hypertension.

Protein tyrosine kinase 2beta (PTK2B) is a member of the focal adhesion kinase family and is activated by angiotensin II through Ca2+-dependent pathways. An evidence exists that PTK2B is involved in cell growth, vascular contraction, inflammatory responses, and salt and water retention through activation of the angiotensin II type 1 receptor. To examine the contribution of PTK2B, we sequenced the PTK2B gene using 48 patients with hypertension, identified 62 genetic polymorphisms, and genotyped six representative single nucleotide polymorphisms in population-based case-control samples from 3655 Japanese individuals (1520 patients with hypertension and 2135 controls). Multivariate logistic regression analysis after adjustments for age, body mass index, present illness (hyperlipidemia and diabetes mellitus), and lifestyle (smoking and drinking) showed -22A>G to have an association with hypertension in men (AA vs. AG+GG: odds ratio=1.27; 95% confidence interval: 1.02-1.57; P=0.030). Another polymorphism, 53484A>C (K838T), in linkage disequilibrium with -22A>G showed a marginal association with hypertension in men (AA vs. AC+CC: odds ratio=1.25; 95% confidence interval: 0.99-1.57; P=0.059). Diastolic blood pressure was 1.6 mmHg higher in men with the AC+CC genotype of 53484A>C than those with the AA genotype (P=0.003), after adjustments for the same factors. These polymorphisms are in linkage disequilibrium with others in a range of 113 kb in PTK2B. The intracellular distribution of the recombinant PTK2B protein and that of the mutant protein with T838 were indistinguishable even after angiotensin II stimulation, both proteins localizing at a focal point in the peripheral area in the cells. Thus, a haplotype in PTK2B may play a role in essential hypertension in Japanese.

Association of genetic polymorphisms of endothelin-converting enzyme-1 gene with hypertension in a Japanese population and rare missense mutation in preproendothelin-1 in Japanese hypertensives.

Endothelin-1 (EDN1), a 21-amino acid peptide, is a potent vasoconstrictor with various pharmacological responses. EDN1 is synthesized from a 212-amino acid precursor protein, preproEDN1, through multiple proteolytic steps. Endothelin-converting enzyme (ECE) cleaves a Trp73-Val74 peptide bond in big-EDN1 to give rise to mature EDN1. In this study, we examined the possible association of genetic variations in ECE1 with hypertension in a general Japanese population and searched for missense mutations in and around the EDN1 polypeptide. We genotyped 5 single nucleotide polymorphisms (SNPs) in the ECE1 gene in 1,873 individuals from a general Japanese population and identified one SNP associated with hypertension in women (rs212528: TT vs. TC+CC: odds ratio=1.40; 95% confidence intervals: 1.04-1.89; p=0.026), after adjusting for confounding factors. The systolic blood pressure in women with the CC genotype was 6.44 mmHg higher than that in those with the TT genotype (p=0.007), after adjusting for the same factors. Next, to identify the missense mutations that may influence the biological activity of EDN1, we sequenced the genomic region that encodes EDN1 in 942 Japanese hypertensive patients. We identified a novel missense mutation, G36R, in one hypertensive patient, but no mutations were observed in EDN1. A gene polymorphism in EDN1, Lys198Asn, has been reported to be associated with hypertension in obese subjects. Taken together, these findings reveal that the EDN-ECE pathway is an important system involved in essential hypertension in Japanese.

Polymorphisms in vitamin K-dependent gamma-carboxylation-related genes influence interindividual variability in plasma protein C and protein S activities in the general population.

gamma-Glutamyl carboxylation, a reaction essential for the activity of vitamin K-dependent proteins, requires the concerted actions of gamma-glutamyl carboxylase (GGCX), vitamin K 2, 3-epoxide reductase complex 1 (VKORC1), and the chaperone calumenin (CALU). We evaluated the contribution of genetic polymorphisms in VKORC1, GGCX, and CALU to interindividual variation in the activities of plasma protein C and protein S. We sequenced these 3 genes in 96 Japanese individuals and geno-typed 9 representative single-nucleotide polymorphisms in 3655 Japanese individuals representative of the general population. The mean activity of protein C in women bearing the GG genotype of GGCX 8016G>A (130.8% +/- 1.5%, n = 156) was significantly greater (P = .002) than that of individuals with either the AG (126.8% +/- 0.7%, n = 728) or the AA (125.4% +/- 0.6%, n = 881) genotype, after adjusting for confounding factors. The GGCX 8016G>A change leads to the substitution of Gin for Arg at amino acid residue 325 (Arg 325 Gln). This effect was comparable to that of a previously defined polymorphism in the protein C promoter. Mean protein S activity was influenced by the VKORC1 3730G>A and CALU 20943T>A genotypes, after adjusting for confounding factors. Thus, polymorphisms in genes involved in the vitamin K-dependent gamma-carboxylation reaction influence interindividual variation in the activities of protein C and protein S in the general population.

Genetic variations of HSD11B2 in hypertensive patients and in the general population, six rare missense/frameshift mutations.

Mutations in the gene encoding 11beta-hydroxysteroid dehydrogenase type 2, HSD11B2, cause a rare monogenic juvenile hypertensive syndrome called apparent mineralocorticoid excess (AME). In AME, defective HSD11B2 enzyme activity results in overstimulation of the mineralocorticoid receptor (MR) by cortisol, causing sodium retention, hypokalemia, and salt-dependent hypertension. Here, we have studied whether genetic variations in HDS11B2 are implicated in essential hypertension in Japanese hypertensives and the general population. By sequencing the entire coding region and the promoter region of HDS11B2 in 953 Japanese hypertensives, we identified five missense mutations in 11 patients (L14F, n = 5; R74H, n = 1; R147H, n = 3; T156I, n = 1; R335H, n = 1) and one novel frameshift mutation (4884Gdel, n = 1) in a heterozygous state, in addition to 19 genetic variations. All genetic variations identified were rare, with minor allele frequencies less than 0.005. Four of 12 patients with the missense/frameshift mutations showed renal failure. Four missense mutations, L14F, R74H, R147H, and R335H, were successfully genotyped in the general population, with a sample size of 3,655 individuals (2,175 normotensives and 1,480 hypertensives). Mutations L14F, R74H, R147H, and R335H were identified in hypertensives (n = 6, 8, 3, and 0, respectively) and normotensives (n = 8, 12, 5, and 0, respectively) with a similar frequency, suggesting that these missense mutations may not strongly affect the etiology of essential hypertension. Since the allele frequency of all of the genetic variations identified in this study was rare, an association study was not conducted. Taken together, our results indicate that missense mutations in HSD11B2 do not substantially contribute to essential hypertension in Japanese.

Mortality in the JACC study till 1999.

BACKGROUND: We have been conducting a cohort study named "the Japan Collaborative Cohort Study (JACC Study) for Evaluation of Cancer Risk sponsored by the Ministry of Education, Science, Sports and Culture of Japan (Monbusho)" since 1988. The aim of this paper is to describe the mortality of our JACC cohort in the follow-up period from 1988 through 1999, to compare it with the mortality, especially cancer deaths, of the Japanese population in the same period and to compare the causes of mortality by district among the cohort. METHODS: We conducted a follow-up study of 110,792 Japanese inhabitants aged 40-79 years in 1988--1990 for about 10 years to the end of 1999. RESULTS: Of 46,465 males, 37,750 (81.2%) were alive, 7,238 (15.6%) were dead and 1,477 (3.2%) had moved out of the study areas. The figures were 57,016 (88.6%), 4,940 (7.7%) and 2,371 (3.7%) among 64,327 females, respectively. The mean follow-up period was 9.9 years. The proportion of cancer deaths by site in our cohort members was almost same as the Japanese population aged 40-79 years old in 1995. Sex-specific standardized mortality ratios of total deaths, all cancer deaths, and most cancers in our cohort were less than 100 in both males and females for total cohort and the cohort by district. CONCLUSION: Our cohort members appeared to be almost the same or slightly healthier and less likely to die from total causes and cancers than the general population.

Discrepancy between clinical and pathological diagnoses of CBD and PSP.

Progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD) are often clinically confused with each other. Moreover, the discrepancy between clinical and pathological diagnoses of CBD and PSP are still controversial. We report here two atypical cases of PSP and CBD. A 73-year old woman was admitted with right hand rigidity, limb kinetic apraxia and cortical sensory loss. Brain atrophy, hypoperfusion and hypometabolism predominantly in the left frontoparietal lobes indicated CBD clinically. Pathological studies revealed neuronal loss and spongy change without ballooned neurons (BN) in the cerebral cortex. Modified Gallyas-Braak (G-B) staining revealed neurofibrillary tangles (NFTs) and tufted astrocytes, indicating pathological diagnosis of PSP. A 75-year-old man admitted with vertical gaze palsy, neck dystonia, parkinsonism and dementia. Atrophy of the frontal lobes and tegmentum of the midbrain and symmetrical frontal hypoperfusion in SPECT indicated PSP. However, neuronal loss and BN in the frontal lobes and clusters of astrocytic plaques indicated CBD pathologically. The G-B staining was useful for differentiating between CBD and PSP, but our atypical cases bring up a new issue about differential diagnosis of CBD and PSP.

Stroke subtypes and lesion sites in Akita, Japan.

Stroke patients who underwent computed tomography (CT) were enrolled in a stroke registry in Akita, Japan, which comprised 7288 first-ever stroke cases during 1999 to 2001. Differences in age and sex were evaluated with respect to type-specific incidences and lesion sites. The incidence increased with age, except for subarachnoid hemorrhage (SAH) in men after age 40. The incidence of intracerebral hemorrhage (IH) and cerebral infarction (CI) was higher in men than in women, whereas that for SAH was lower. The mean age of putaminal hemorrhage was lower than that of thalamic hemorrhage, and the mean age of cortical infarction was higher than that of CI in perforator regions, the cerebellum, and the pons. In subjects age 70 years and older, the proportion of thalamic hemorrhage in IH was larger in women than in men; for those age 50 years and older, the proportion of cortical infarction in CI was larger in men than in women. The proportions of anterior communicating artery aneurysms in men and internal carotid artery aneurysms in women were largest in SAH for all age groups. In conclusion, thalamic hemorrhage was most common in elderly women and cortical infarction was most common in middle-aged and elderly men. The feature of SAH occurring at a higher incidence in women than in men, with a sexual difference in aneurysmal distribution, was also observed.

A novel missense mutation, F826Y, in the mineralocorticoid receptor gene in Japanese hypertensives: its implications for clinical phenotypes.

A gain-of-function mutation resulting in the S810L amino acid substitution in the hormone-binding domain of the mineralocorticoid receptor (MR, locus symbol NR3C2) is responsible for early-onset hypertension that is exacerbated in pregnancy. The objective of this study was to test whether other types of missense mutations in the hormone-binding domain could be implicated in hypertension in Japanese. Here, we screened 942 Japanese patients with hypertension for the S810L mutation in exon 6 in the MR. We did not identify the S810L mutation in our hypertensive population, indicating that S810L does not play a major role in the etiology of essential hypertension in Japanese. However, we identified a novel missense mutation, F826Y, in three patients in a heterozygous state, in addition to four single nucleotide polymorphisms, including one synonymous mutation (L809L). The F826Y mutation is present in the MR hormone-binding domain and might affect the ligand affinity. The F826Y mutation was also identified in 13 individuals (5 hypertensives and 8 normotensives) in a Japanese general population (n=3,655). The allele frequency was 0.00178. The frequencies of the F826Y mutation in the hypertensive population (3/942) and in the hypertensive group (5/ 1,480) and the normotensive group (8/2,175) in the general population were not significantly different, suggesting that this mutation does not greatly affect hypertension. Although it is unclear at present whether or not the F826Y mutation makes a substantial contribution to the mineralocorticoid receptor activity, this missense mutation may contribute, to some extent, to clinical phenotypes through its effects on MR.

Relationship between abdominal visceral fat and lacunar infarcts in Japanese men.

BACKGROUND: The relationship between abdominal visceral fat accumulation and lacunar infarcts has not been previously investigated in Japanese men. METHODS AND RESULTS: The subjects were 637 middle-aged (40-64 years) and 222 elderly (65-79 years) men who participated in a health checkup program from 1999 to 2003. The association between lacunar infarcts identified by magnetic resonance imaging and cardiovascular risk factors, including abdominal visceral fat accumulation evaluated by computed tomography, was examined. The prevalence of lacunar infarcts was 4.9%. Hypertension was associated with lacunar infarcts among both the middle-aged men [age-adjusted odds ratio (OR)=2.9 (95% confidence interval (CI): 1.1-7.8)] and the elderly men [OR=5.1 (95%CI: 1.4-19.0)]. Abdominal visceral fat accumulation was slightly associated with lacunar infarcts among middle-aged men, but not among elderly men: OR in the highest (>or=117 cm(2)) vs lowest (or=143 cm(2)) was still slightly associated with lacunar infarcts after adjustment for age, hypertension, drinking and smoking among middle-aged men [OR=2.7 (95%CI: 0.8-9.1)]. CONCLUSIONS: This cross-sectional study suggests that abdominal visceral fat accumulation is a possible risk factor of lacunar infarcts, in addition to hypertension, in middle-aged Japanese men.

Diffusion-weighted MRI in evaluation of transient ischemic attack.

Diffusion-weighted magnetic resonance imaging (DWI) is a sensitive diagnostic tool for detecting recent ischemic lesions in patients with transient ischemic attacks (TIAs), but the interpretation of the presence or absence of DWI abnormalities in TIA patients still remains controversial. To elucidate the pathophysiology underlying those lesions, we analyzed DWI abnormalities in patients with recent TIAs. Based on 45 consecutive patients with TIAs who underwent DWI within 10 days of onset, demographic data and clinical manifestations were analyzed in relation to the DWI abnormalities. According to the method utilized in the Oxfordshire Community Stroke Study, clinical manifestations were classified into classical lacunar syndrome and non-lacunar symptoms. Based on the vascular distributions of ischemic lesions, the DWI abnormalities were classified into small-vessel and large-vessel lesions. DWI abnormalities were detected in 14 (31%) of 45 TIA patients. Seven (50%) of 14 DWI-positive patients had occlusive vascular lesions on intracranial magnetic resonance angiography, while only 5 (16%) of 31 DWI-negative patients had occlusive lesions (P < .05). No other demographic or clinical features, including risk factor and presence of cardiac disease, differed significantly between the DWI-positive and DWI-negative patient groups. Four (46%) of 9 DWI-positive patients who had a classical lacunar syndrome also showed small-vessel lesions on DWI, whereas all 5 patients who had non-lacunar symptoms showed large-vessel lesions. We concluded that although DWI abnormalities were detected in only one third of our TIA patients, DWI abnormalities were closely related to intracranial vascular occlusive lesions. The combination of DWI and MRA was useful for detecting large-artery lesions in patients displaying a classical lacunar syndrome.