Association of the missense Glu298Asp variant of the endothelial nitric oxide synthase gene with myocardial infarction.

OBJECTIVES: We examined the possible association between the missense Glu298Asp variant of the endothelial nitric oxide synthase (eNOS) gene and myocardial infarction (MI). BACKGROUND: Endothelium-derived nitric oxide (NO) plays a key role in the regulation of vascular tone. Recently, we reported that a missense Glu298Asp variant in exon 7 of the eNOS gene is a possible genetic factor involved in the pathogenesis of coronary spasm. Endothelium-derived NO also has vasoprotective effects by suppressing platelet aggregation, leukocyte adhesion and smooth muscle cell proliferation. METHODS: We screened 285 patients with an MI and 607 control subjects in Kumamoto Prefecture, Japan. Genotypes were determined by polymerase chain reaction-restriction fragment-length polymorphism analysis. RESULTS: The frequency of the missense Glu298Asp variant was significantly higher in the MI group than in the control group (21.1% vs. 13.3%, p = 0.003, odds ratio 1.73 for the dominant effect of the eNOS T allele). Multiple logistic regression analysis showed that the missense Glu298Asp variant was an independent risk factor for MI, as was diabetes mellitus, hypertension, cigarette smoking, hypercholesterolemia and body mass index. CONCLUSIONS: There was a significant association of the missense Glu298Asp variant of the eNOS gene with MI. This marker-disease association may be due to the impaired effects of NO on the cardiovascular system: dysregulation of vascular tone, platelet aggregation and leukocyte adhesion and smooth muscle cell proliferation, all of which promote coronary atherosclerosis and thrombosis.

Primary biliary cirrhosis. A patient with adverse reactions to tiopronin and autoimmune hemolytic anemia with reticulocytopenia.

A 32-year-old woman with asymptomatic primary biliary cirrhosis had autoimmune hemolytic anemia associated with reticulocytopenia and thrombocytopenia despite an intensely erythroid bone marrow. Her anemia was successfully treated with oral prednisolone and intravenous pulse methylprednisolone, with a rapid response of reticulocytosis and sustained erythrocytosis. Tiopronin therapy was later initiated and resulted in fever, rash, exacerbation of the liver disease, and positive direct and indirect antiglobulin tests.

Clinical characteristics of Japanese men with glucokinase gene beta-cell promoter variant.

OBJECTIVE: To study the association between a variant at the position of -30 of beta-cell-specific promoter of the glucokinase gene and glucose tolerance in the Japanese general population and to assess the clinical characteristics of subjects with the variant. RESEARCH DESIGN AND METHODS: The genotype of 657 Japanese men aged 51.0 +/- 8.8 years (mean +/- SD) was analyzed by an allele-specific assay using polymerase chain reaction-restriction fragment length polymorphism. RESULTS: The variant allele frequency was 0.188 in subjects with normal glucose tolerance, 0.211 in subjects with impaired glucose tolerance, and 0.176 in diabetic subjects. In subjects with fasting plasma glucose levels <140 mg/dl, homozygous subjects for the promoter variant had significantly higher plasma glucose levels 60 min after oral glucose administration when compared with subjects without the variant allele. A cross-sectional analysis showed age-related elevation of basal glucose levels only in subjects without the promoter variant. Individuals heterozygous for the variant had significantly lower levels of HDL cholesterol than normal subjects. HDL cholesterol values were lower in homozygous people than in normal and heterozygous subjects, although the differences were not statistically significant. CONCLUSIONS: The beta-cell promoter variant in homozygous state was associated with impaired glucose tolerance, but not with diabetes, and low HDL cholesterol levels in Japanese men. It is unlikely that the glucose intolerance associated with the promoter variant is progressive with age.

Codon 972 polymorphism of the insulin receptor substrate-1 gene in impaired glucose tolerance and late-onset NIDDM.

OBJECTIVE: To assess the relevance of a Gly-->Arg substitution in codon 972 of the insulin receptor substrate-1 gene in impaired glucose tolerance (IGT) and NIDDM. RESEARCH DESIGN AND METHODS: The genotype of 1,106 Japanese subjects consisting of 310 subjects with NIDDM, 305 subjects with IGT, and 491 normal control subjects was analyzed by an allele-specific assay using polymerase chain reaction and restriction fragment length polymorphism. RESULTS: The frequency of the variant allele was not different between subjects with NIDDM (0.021) and normal control subjects (0.020). However, subjects with IGT showed a significantly higher prevalence of the variant allele (0.041, P = 0.027). We found two homozygous individuals for the variant; both had IGT with mild insulin resistance. The allelic frequency tended to be lower in normal control subjects aged > 50 years than in younger control subjects. Conversely, in the subjects with IGT or NIDDM, the Gly972Arg substitution was more frequently found in subjects aged > 50 years. Furthermore, NIDDM patients with the variant allele had older ages of diagnosis than patients without the variant. CONCLUSIONS: The codon 972 variant may be associated with IGT and a subset of late-onset NIDDM in the elderly Japanese population.

Polymorphism in the 5'-leader cistron of the beta2-adrenergic receptor gene associated with obesity and type 2 diabetes.

We screened the 5'-untranslated region of the beta2-adrenergic receptor gene from 40 obese subjects by the PCR-direct sequencing technique. Two polymorphic sites were identified; a T-->C substitution at -47 and a T-->C substitution at -20. We further analyzed the association of the polymorphisms with obesity in 574 subjects by PCR and restriction digestion. The substitution at -47 was in tight linkage disequilibrium with that at -20. The polymorphisms were also in linkage disequilibrium with codon 16 and codon 27 polymorphisms. Subjects carrying the -47C/-20C allele had greater body mass index (25.5+/-4.5 vs. 24.4+/-4.1 kg/m2, p=0.007) and higher serum triglyceride levels (166+/-160 vs. 139+/-95 mg/dl, p=0.015) than -47T/-20T homozygotes. The variant allele frequency was significantly higher in obese subjects than in non-obese subjects (0.18 vs. 0.11, p=0.0026). Furthermore, an increased frequency of the variant allele was shown in diabetic patients compared with non-diabetic subjects (0.19 vs. 0.11, p=0.0005). The association may be attributable to the greater proportion of diabetic patients in the obese group. The exchange at -47 may alter the expression level of the beta2-adrenergic receptor gene, because the nucleotide substitution at -47 results in a Cys-->Arg exchange at the C terminal of the leader peptide. The -47C/-20C allele may be associated with genetic predisposition to obesity and obesity-related metabolic disorders.

Analysis of trinucleotide-repeat combination polymorphism at the rad gene in patients with type 2 diabetes mellitus.

A combined (GTT)n (ATT)n trinucleotide-repeat polymorphism designated as RAD1 has been identified at intron 2 of the rad gene on chromosome 16q. An association between the total length of the RAD1 locus and type 2 diabetes has been shown in white American subjects, but not in Finns. We genotyped 115 Japanese patients with type 2 diabetes and 114 nondiabetic control subjects at the RAD1 locus by the direct sequencing method, and found 16 RAD1 alleles composed of various combinations of GTTs and ATTs. Allele 14 consisting of four GTTs and seven ATTs accounted for the majority in both control subjects and diabetic patients, suggesting that RAD1 polymorphism is not a major genetic component for susceptibility to common forms of diabetes in the Japanese. There was no significant association between total repeat length and diabetes. However, the frequency of minor alleles containing five GTTs or three GTTs was significantly higher in diabetic patients versus nondiabetic subjects (4.8% v 0.9%, P = .012). Thus, genetic variability at the rad gene in linkage disequilibrium with RAD1 could be associated with a predisposition to type 2 diabetes in the Japanese population.

Follow-up studies on the distribution of haemoglobin levels in female farm workers.

Haemoglobin concentrations in about 1000 women agricultural workers in Japan were measured every year, except in 1972, during the period 1967-77. Improvements were noted in the course of this investigation, and these were predominantly associated with the fact that those in the study community began to pay attention to the problem of low haemoglobin levels and to improve their diet, with an increase in daily food intake, particularly of animal protein and iron. In addition, a marked decrease in anaemia caused by hookworm also played an important role. In this paper, the change in haemoglobin concentration during the period of study are described and the aetiology is discussed.

Beta 3-adrenergic receptor gene polymorphism is not a major genetic determinant of obesity and diabetes in Japanese general population.

To assess the contribution of a replacement of Trp at codon 64 of beta 3-adrenergic receptor by Arg to fat distribution and metabolic disturbances in Japanese general population, we examined the missense mutation in 1122 persons consisting of 817 men aged 50.0 +/- 8.9 years and 305 women aged 50.8 +/- 8.5 years in Kyushu, Japan. The incidence of Arg64 allele was 0.21; no age-dependent decrease of the allele frequency was observed, suggesting that the mutation was not associated with early mortality. The genotype was not significantly correlated with body mass index or the thickness of visceral fat estimated by ultrasonography. Glucose tolerance and glucose-induced insulin secretion were not significantly different among subjects with Trp/Trp, Trp/Arg and Arg/Arg at codon 64. Although in obese persons the ratio of heterozygotes for the mutation tended to be higher in subjects with impaired glucose tolerance than in subjects with normal glucose tolerance, the tendency was not observed in non-obese persons. Furthermore none of 39 non-obese individuals homozygous for the mutation was diabetic, whereas two out of six obese homozygous persons were diabetic. These observations suggest that the missense mutation may not be a main determinant of obesity in populations taking low fat/low energy Japanese-style diet and it may not be deleterious at least in non-obese individuals.

Glycemic index of single and mixed meal foods among common Japanese foods with white rice as a reference food.

OBJECTIVE: The objectives were to examine the feasibility of using white rice as a reference food in the study of glycemic index (GI) and to examine the GI values of both single and mixed meal foods among rice species, processed rice products, beans, and dairy products. DESIGN: Subjects were served with 50 g carbohydrate content of white rice at least two times (maximum three times) and test food once after separate overnight fasts. Capillary blood glucose measurements were carried out before and during 120 min after each food load. SETTING: The study was carried out in an outpatient setting. SUBJECTS: A total of 58 (38 females and 20 males) nondiseased subjects, mean aged 37 y and mean BMI 22 kg/m(2) were included. RESULT: The correlation between incremental area under curve of white rice and glucose was r=0.853 (n=10, P <0.0001) and white rice was considered suitable to be used as a reference food. Among mixed meal foods, the combination of carbohydrate foods with vinegar, dairy products, and bean products significantly decreased the GI value of white rice of 20-40%. The reduction of GI occurred whether the foods were taken together, before or after rice intake. GI of noodles such as udon, soba, and spaghetti showed low GI values. CONCLUSION: White rice could be used as a reference food in determining GI values of foods. A total of 32 single and mixed meal Japanese common food products were examined for their GI values.

Early detection of renal cell carcinoma by ultrasonographic screening--based on the results of 13 years screening in Japan.

Abdominal ultrasonographic (US) screening of 219,640 persons has been performed in the past 13 y, and 723 (0.33%) cases of malignant neoplasm were detected. Renal cell carcinoma (RCC) was detected in 192 cases (0.09% of the examinees). In almost all cases of RCC, no symptoms were evident and no abnormalities were detected in the blood chemistry tests or urinalyses. A total of 189 cases (98%) were resected curatively, and 38% of the tumors were less than 25 mm in size (T1). With respect to pTNM classification, 35% were pT1 and 52% were pT2. No metastasis to the lymph nodes or other organs was found in any case. The cumulative survival rate for cases resected was 97% at 5 y, and 95% at 10 y. Regarding US features of RCC, the internal echo pattern of half of T1 tumors showed homogeneous and hyperechoic, and became heterogeneous as they grew. Other notable US findings in cases of RCC were marginal hypoechoic zone (29%), anechoic component in the tumor (23%), and protrusion from the kidney (85%, 71% of the T1 tumors). US screening is useful for detection of RCC in the early stage. However, to detect small tumors, it is very important to know well the US features of RCC. For cost-effectiveness analysis, it is more effective to examine, not only the kidney, but other abdominal organs. It is expected that many other abdominal cancers, such as hepatocellular carcinoma, gallbladder cancer, pancreatic cancer, and so on, could be found in the early stage by broad implementation of US screening.

Lifestyle change improves individual health and lowers healthcare costs.

In 1996 the Japanese Ministry of Health and Welfare discontinued the use of the term "Adult onset diseases" used for almost 40 years, and replaced it by "Lifestyle-related diseases"; a meaningful change. Lifestyle is one of the greatest problems facing Japan today. Considering the rapid aging of Japanese society and the continuing increase in medical costs, it is important for all citizens to be concerned not only about their own health but also about the impact on the financial health of the nation as a whole. Our data indicate that the greatest determining factor for increased healthcare costs is the percentage of patients hospitalized, and the variable with the greatest influence on this is the relative number of hospital beds available. On the other hand, the variable with the greatest inhibitory effect on the growth of healthcare costs is the availability of guidance to improve lifestyle factors.

Significance of measuring the quality of life in health evaluation.

To evaluate the QUIK-R, a revised self-completed questionnaire for the quality of life (QOL), and to clarify the significance of measuring QOL in health evaluations, the results of 1,017 office workers were analyzed. The QUIK-R showed a high reliability and a high validity. There were significant correlations between the total score and the subject's age, sex, smoking habit, sleeping time, and exercise. Forty-two subjects with a good QUIK-R score had poor physical examination results. Our results show the usefulness of the QUIK-R and the importance of measuring the QOL in health evaluations.

Prevalence and conditions of urinary incontinence among the elderly.

In Japan, elderly disorders and diseases have markedly increased in recent years, because of rapid aging and an increasing number of older persons. The situation is creating serious social and community problems. These disorders, particularly dysuria and urinary incontinence (UI), disturb the quality of life (QOL) in latelife. Few reports on UI have been published, but precise investigation into the community level remains to be made. Our presentation is the development, implementation and evaluation of elderly UI in Kumamoto Prefecture. This study includes 2,304 people (male: 856, female: 1,448), over 65 years of age, living in two different communities; one is an urban (K) and the other is a typical rural area (S). The rate of UI was in homebound elderly persons, male: 4.7%, female: 11.3%, and in nursing home residents, male: 16.2%, female: 23.2%. The condition of UI was: almost Urinary Urgency in male (61.5%), and Stress Incontinence (such as, caused by coughing, sneezing, and exercise) in female (46.3%). The influence of UI on the activity of daily life was investigated. Most of the male cases were giving concerns for family and community. In contrast, females hesitated to participate in group excursions and outdoor exercise, and had a tendency to live alone or indoors. However, most persons (81.5%) with UI did not visit a physician. From this investigation, we conclude that a community health care program and public support system are essential for proper understanding and solution of the elderly UI problem.

[A case of advanced gastric cancer responsive to intratumoral OK-432 injection].

An 83-year-old patient with advanced gastric cancer was given repeated endoscopic administration of OK-432 combined with systemic immunochemotherapy. The cancerous lesion gradually regressed following the treatment, and remarkable degeneration was observed in many tumor cells. Detailed immunohistochemical examination revealed that Leu 7-positive cells had markedly infiltrated the cancerous tissue. This finding suggests that NK cells may play an important role in tumor regression induced by intratumoral OK-432 therapy.

[The reliability and validity of a new self-completed questionnaire (QUIK)].

In order to evaluate the reliability and validity of a self-completed questionnaire (QUIK), devised to measured QOL, we examined the QUIK scores of elderly who visited the Kumamoto Health Administrative Center for a medical check-up in March 1994. The QUIK questionnaire, which is a close-ended and disease non-specific questionnaire, covered four domains such as physical functioning, emotional adjustment, interpersonal relationships, and attitudes toward life, interacting reciprocally. The mean and standard deviation on QUIK were much better in terms of total score (5.1 +/- 5.4), for each domain score in comparison with the patient group, and even in comparison with the non-patient group. The distribution of total scores on QUIK were as follows: excellent 15%, good 35%, fair 36%, poor 11%, very poor 2% and grossly impaired 0% according to a six-tiered rating scale. The internal consistency in terms of total score was alpha = 0.86. Very close correlation were seen among score, each domain score and satisfaction, being healthy and present state of feeling. If the cut-off points of total score were set between 9 and 10, the sensitivity were 0.65, specificity was 0.65 for the age index, sensitivity 1.00, validity 0.29 for the satisfaction index, while, sensitivity was 0.85 and validity 0.48, for the feeling index. There was a very close reciprocal correlation among the four domains, except for the relation between physical functioning and interpersonal relationship using multiple regression analysis. Further, significant correlations were obtained between the score in each domain and the score based on subtracting each domain score from the total score.