小谷 和彦

OBJECTIVE: It was the aim of this study to investigate whether there is any relationship between oxidative stress, as assessed by the diacron reactive oxygen metabolite (d-ROM) test, and carotid atherosclerosis among hypercholesterolemic patients. SUBJECTS AND METHODS: A well-defined group of patients with type II hypercholesterolemia (n = 81, mean age 59 years) was studied to observe the correlation between the levels of serum d-ROMs and carotid artery intima-media thickness (IMT) using B-mode ultrasound, in relation to the traditional atherosclerotic risk factors (age, sex, smoking, body mass index, blood pressure, glucose and lipid panels). RESULTS: The mean level in low-density lipoprotein cholesterol (LDL-C) in this population was 4.45 mmol/l, d-ROMs were 323.2 Carr U, and IMT was 0.91 mm. A multiple regression analysis revealed a positive and significant correlation between IMT and d-ROMs (β = 0.27, p < 0.05), along with age and LDL-C. CONCLUSION: These results indicate that the increased oxidative stress levels using the d-ROM test, independent of aging and increased LDL-C levels, may be associated with carotid atherosclerosis even in hypercholesterolemic patients.

Hypertension (HT) and C-reactive protein (CRP) are risk factors of cardiovascular diseases (CVD). The current study's purpose was to investigate the relationship between serum CRP levels and daily lifestyles, including physical activity, in Japanese HT patients. Lifestyle factors, blood pressure (BP), blood cholesterol, glucose, and CRP were measured in a total of 312 HT patients (153 men/159 women, mean age: 62.6 y). Women with physical activity of ≥ 1 time/week showed significantly lower CRP levels than those without it (p < 0.05). The data suggest that regular physical activity could reduce the CRP levels in HT patients, thereby maybe preventing CVD.

The uncoupling protein-1 (UCP1) gene is of major importance for regulation of body weight and lipid/lipoprotein metabolism. Our cross-sectional study has shown that subjects with the G/G genotype of the -3826 A/G polymorphism in the UCP-1 gene have higher levels of serum high-density lipoprotein cholesterol (HDL-C) than those with other genotypes. Low circulating HDL-C level has been regarded as a major atherosclerotic risk factor. We therefore investigated whether the -3826 A/G polymorphism affects the obesity- and lipid-related parameters during a low-calorie diet (LCD) intervention. In 32 obese women (49.9 +/- 8.4 years of age), anthropometric, physiological and biochemical characteristics were measured before and after a 2-month LCD treatment, which restricted each subject to the same energy intakes, such as 5,120 kJ/day. The -3826 A/G polymorphism was detected using a PCR-restriction fragment-length polymorphism method. There were 6 subjects with the A/A genotype, 15 with the A/G genotype and 11 with the G/G genotype. The LCD intervention decreased weight (P < 0.001) and serum HDL-C levels (P < 0.05) in all subjects. There was no difference in the levels of change in weight, nutrient intake, physiological measurements in energy expenditure, and fat oxidation between subjects with and without the G allele. In contrast, the degree of the reduction in the HDL-C levels was significantly smaller in subjects with the G allele than those without the G allele. These results suggest that the G allele at -3826 in the UCP1 gene may ameliorate the reduction in serum HDL-C levels in obese women during LCD.

BACKGROUND: Lifestyle modification improves the pathophysiology of lipid disorder, leading to the development of cardiovascular disease (CVD). While oxidized low-density lipoprotein (oxLDL) may be involved in this mechanism, various oxLDL measurements have recently been developed and therefore further detailed studies are called for in this area. Our aim was to investigate the effects of lifestyle modification on serum amyloid A-LDL (SAA-LDL), a new oxLDL, in subjects with primary lipid disorder. METHODS: A total of 141 asymptomatic subjects (women/men=100/41, mean age 57.6 years) with>or=1 lipid abnormality (circulating high levels of low-density lipoprotein cholesterol [LDL-C] and triglyceride [TG] or low levels of high-density lipoprotein cholesterol [HDL-C]), who completed a 6-month lifestyle modification program in combination with diet and exercise, were analyzed. In the pre- and post-intervention, the metabolic variables including SAA-LDL were assessed. RESULTS: During our intervention, the body mass index, blood pressure, LDL-C, TG, glucose and SAA-LDL significantly decreased, while HDL-C significantly increased. Multiple regression analysis revealed that the change levels of TG (positive) and HDL-C (inverse) were significantly and independently correlated to those of SAA-LDL. CONCLUSIONS: These results suggest that SAA-LDL may contribute to the link between lipid disorder and the development of CVD, and that the application of SAA-LDL measurements may be useful for the assessment of the risk of CVD as a biochemical marker.

BACKGROUND: C-reactive protein (CRP) is an important risk factor for cardiovascular disease. Furthermore, it has been reported that levels of CRP are increased in patients with obstructive sleep apnea (OSA). The aim of this study was to examine the effects of long-term therapy with nasal continuous positive airway pressure (nCPAP) on CRP levels and to investigate whether compliance with nCPAP therapy more effectively attenuated markers of systemic inflammation in patients with OSA. METHODS AND RESULTS: Fifty-five patients (mean [+/- SEM] age, 55 +/- 2 years; 44 male patients, 11 female patients) with newly diagnosed moderate-to-severe OSA (apnea-hypopnea index > 20 events/h) were studied before and after 6 months of nCPAP treatment. There was a significant reduction in CRP levels after nCPAP therapy (before nCPAP therapy, 0.23 +/- 0.03 mg/dL; after nCPAP therapy, 0.17 +/- 0.02 mg/dL; p < 0.01). Additionally, we divided these patients into two groups based on adherence to nCPAP therapy. A group of patients using nCPAP > 4 h/d and > 5 d/wk were designated as the good compliance group. The decrease in CRP concentration was significant (before nCPAP therapy, 0.23 +/- 0.04 mg/dL; after nCPAP therapy, 0.16 +/- 0.03 mg/dL; p < 0.05) in the good compliance group but not in the poor compliance group (before nCPAP therapy, 0.24 +/- 0.05 mg/dL; after nCPAP therapy, 0.20 +/- 0.05 mg/dL; p = 0.21). Furthermore, we divided those patients into a high CRP group (>/= 0.2 mg/dL) and a normal CRP group (< 0.2 mg/dL) before nCPAP therapy. The significant decrease in CRP levels in the good compliance group was evident only in those patients with an initially elevated CRP level (before nCPAP therapy, 0.48 +/- 0.08 mg/dL; after nCPAP therapy, 0.29 +/- 0.06 mg/dL; p < 0.05). CONCLUSION: Appropriate use of nCPAP in patients with OSA may be required to decrease elevated CRP levels, with possible implications for cardiovascular morbidity and mortality.

BACKGROUND: Hemorheology plays an important role in the development of cardiovascular disease. The Micro Channel array Flow Analyzer (MC-FAN) (Hitachi Haramachi Electronics Co., Ltd., Bentencho, Japan) is currently considered a useful new device to analyze hemorheology. However, the relationships between various lifestyle habits and hemorheology, especially using MC-FAN, have still not been thoroughly investigated. HYPOTHESIS: The study was undertaken to determine whether there could be some correlations of lifestyle factors to hemorheology by MC-FAN. METHODS: A total of 250 asymptomatic Japanese subjects (male:female = 100:150; mean age = 49.7 y) without any medication were enrolled in the present study. Hemorheology was assessed by the whole blood passage time (WBPT) and was corrected by the saline passage time using MC-FAN. Subjects' lifestyle factors, such as smoking habits, alcohol habits, and physical activity, were self-reported. RESULTS: Partial correlation analysis for WBPT, after controlling for age, gender, hematocrit, white blood cell count, body mass index, blood pressure, blood biochemical measures, and all lifestyle habits, revealed a significant and inverse correlation between alcohol habits of 1-3 go (amount of alcohol intake) and WBPT (r = - 0.191, p = 0.003), in addition to a significant positive correlation between serum low-density lipoprotein (LDL) cholesterol and WBPT. CONCLUSIONS: These data suggest that alcohol habits may beneficially affect hemorheology by MC-FAN, expanding the protective effect of light-to-moderate alcohol consumption against cardiovascular disease.

Nine clones of non-pathogenic streptococci were isolated from the pharynges of seven healthy subjects, and grown on sheep blood agar plates with a hemolysis or gamma hemolysis, then cultured in LB broth for 16 hrs. Purified streptolysin O (SLO) purchased from Sigma Chemical Co. (Sigma-SLO), SLO antigen as a latex agglutination reagent from A company (A-SLO) and supernatants from four culture media were electrophoresed on 12% SDS-polyacrylamide gel and transferred to PVDF membranes. Immunological analyses of antibodies against SLO in healthy sera and proteins in culture medium demonstrated that healthy sera contained an antibody recognizing Sigma-SLO, A-SLO and a protein of the same size as SLO (SLO-like protein) in culture medium. These findings suggest that healthy subjects develop an antibody directed against SLO-like protein produced by non-pathogenic streptococci, and that this antibody cross-reacts with Sigma-SLO and A-SLO. Using DNA from Streptococcus pyogenes and non-pathogenic streptococci, the SLO gene and SLO-like protein gene were analyzed by direct sequencing with oligonucleotide primers designed to cover no. 74 to approximately 1900 of the SLO gene. There were three different bases resulting in amino acid substitution between the SLO gene and SLO-like protein gene, namely 101Lys (AAA) of SLO to Asn (AAT), 175Met (ATG) to Arg (AGG) and 185Asp (GAT) to Asn (AAT). Remaining 560 residues of 563 amino acids constituting SLO-like protein were homologous to SLO. Non-pathogenic streptococci on the pharynges of healthy subjects produce an SLO-like protein composed of amino acids similar to those of SLO, which induces an antibody against this SLO-like protein in serum. It is likely that an antibody against SLO-like protein in healthy sera cross-reacts with SLO and causes a pseudo-positive reaction on ASO measurement by the latex agglutination method using SLO antigen.

BACKGROUND: Polymorphism of Trp64Arg in the beta(3)-adrenergic receptor (beta(3)-AR) gene may play a critical role in lipid and lipoprotein metabolism by mediating lipolysis and thermogenesis. Since the frequency of Arg alleles of the beta(3)-AR gene is generally low among many populations, studies on the Arg/Arg genotype in relation to lipid and lipoprotein metabolism are required in countries such as Japan which has a relatively high frequency of the Arg allele. METHODS: We genotyped 275 clinically healthy Japanese (male/female, 134/141, mean 45.7 years) without medication for beta(3)-AR gene polymorphism by polymerase chain reaction-restriction fragment length polymorphism analysis, and investigated the effects of the gene polymorphism on clinical parameters including body mass index (BMI), blood pressure and serum lipid and lipoprotein concentrations. RESULTS: The genotype frequencies were: Trp/Trp, 68.0%; Try/Arg, 28.0% and Arg/Arg, 4.0%, with an allele frequency of 0.18 for Arg64. When subjects were divided into three groups according to the genotype, a significant increase of serum LDL-cholesterol (LDL-C) concentration in the Arg/Arg group (3.48 +/- 1.59 mmol/L) was observed when compared with those of the Trp/Trp and Arg/Trp groups (3.15 +/- 0.80, 3.25 +/- 0.92 mmol/L, respectively). Genotype differences did not show any significant effect on other parameters. Spearman's rank correlation demonstrated a significant relationship between LDL-C concentrations and the number of Arg alleles, age and BMI. Multiple regression analysis, using LDL-C concentration as a criterion variable and some factors including beta(3)-AR gene polymorphism as explanatory variables, revealed that the number of Arg alleles was a significant and independent factor for LDL-C concentrations, along with age and BMI. CONCLUSIONS: These findings suggested a role of the beta(3)-AR gene polymorphism in regulating lipid and lipoprotein metabolism, showing small but significant effects on elevated LDL-C values in subjects with Arg/Arg, but not Trp/Arg and Trp/Trp genotypes.

Sleeplessness can be linked to hypertension and its consequent complications such as cardiovascular disease. We investigated the cross-sectional relationship between blood pressure (BP) and self-reported sleep status among 227 healthy normotensive Japanese females (mean age: 47.0 years) in a clinical setting. Multiple logistic regression analysis for the highest quartile of diastolic BP (DBP) showed the body mass index (BMI) (odds ratio=1.26 [95% confidence interval=1.12-1.41]) and sleep status (sometimes lack: 2.19 [1.00-4.79], poor: 2.82 [1.17-6.80]) as significantly associated factors. The same analysis for systolic BP or pulse pressure showed that both age and BMI were significant factors and sleep status was not associated. A poorer sleep status may contribute to elevated DBP in healthy normotensive females.

Gamma-glutamyl transferase (GGT) is an enzyme present in serum and on most cell surfaces and serves as an oxidative stress marker. Although serum GGT is associated with hypertension development, little data are available on the associations between GGT and hypertension among populations with diabetes mellitus (DM). Our aim was to investigate the potential association between the changes in systolic or diastolic blood pressure (SBP/DBP) and the GGT level in type 2 DM subjects, in comparison with non-DM subjects. In 179 non-DM and 177 DM subjects, SBP/DBP, body mass index (BMI), fasting plasma glucose, serum asparate aminotransferase, alanine aminotransferase and GGT were measured at the baseline and after a 1-year period. Between these 2-measurement points, in non-DM subjects, SBP and DBP levels were significantly increased, while GGT tended to increase. In contrast, in DM subjects, the mean levels of SBP, DBP and GGT remained unchanged. Multivariate analysis revealed that in non-DM subjects the degree of increase in SBP was significantly and positively correlated to that of GGT (beta = 0.165), along with age and BMI. Likewise, the increase in DBP was correlated to that of GGT in non-DM subjects (beta = 0.170). In contrast, in DM subjects, the degree of increase in SBP was significantly correlated to that of only GGT (beta = 0.166). These results suggest that the presence of DM may attenuate the effects of GGT on DBP.

A close relationship between coffee intake and certain metabolic disorders is known. Caffeine, one of coffee components, can increase energy expenditure (EE), but there are considerable individual differences in the caffeine effects on EE, and the causes have not been fully established in humans. The Arg allele in the beta(3)-adrenergic receptor gene (beta(3)-AR), a marker for obesity-related traits, may be a contributor to individual variations in EE. This study investigated the effect of the Arg allele of beta(3)-AR on caffeine-induced increases in EE. In 44 healthy young women (21 +/- 1 years), physical characteristics, blood pressure, biochemical profiles and dietary nutritional intake were measured. A caffeine-loading test was conducted at a dosage of 4 mg per body weight (kg). EE was measured using an indirect open-circuit calorimeter for a 10-min period before, and at 30 min and 60 min after the caffeine-loading test. The beta(3)-AR Trp64Arg polymorphism was detected with a PCR-restriction fragment length polymorphism method. The frequency of the Arg allele was 24%. The distribution of the Trp/Trp, Trp/Arg, and Arg/Arg genotypes was 58%, 36%, and 6%, respectively. At the baseline, subjects with the Arg/Arg genotype had a significantly lower EE level than those with the Trp/Trp or Trp/Arg genotype. After the caffeine-loading test, there were caffeine-induced increases in EE in all genotypes, but there were no differences in the levels of increase among the genotypes. These findings suggest that the genotypes of beta(3)-AR Trp64Arg polymorphism might be not associated with caffeine-induced increases in EE levels.

Uncoupling protein 3 (UCP3) is considered to be associated with obesity, given its function in the regulation of energy and lipid metabolism. An increased body mass index (BMI) and a decreased level of high-density lipoprotein cholesterol (HDL-C) are risk factors for cardiovascular disease. The purpose of this study was to investigate whether the UCP3 promoter -55 C/T single nucleotide polymorphism (UCP3 -55 C/T SNP) was associated with obesity according to the criteria for Japanese (BMI > or = 25 kg/m2), BMI, and serum HDL-C levels in the general Japanese population. The subjects, numbering 282 and aged 65 +/- 13 years (mean +/- SD), were recruited through an annual health checkup of residents of Mima city, Tokushima, in Japan. Body mass index, blood pressure, biochemical indexes including lipid, and lipoprotein profiles were measured. The UCP3 -55 C/T SNP was determined with a fluorescence-based allele-specific DNA primer assay system. The frequency of the -55 T allele was 30.0%. Subjects with the T/T genotype had significantly higher HDL-C levels than those with the C/C genotype or the C/T genotype. Furthermore, subjects with the T/T genotype had a significantly lower BMI than those with the C/C genotype. A multivariate analysis revealed that the -55 T allele was a significant independent variable contributing to the variance in HDL-C levels and BMI. The T/T genotype was associated with a lower prevalence of obesity than the C/C and C/T genotypes, with an odds ratio of 0.358 (95% confidence interval, 0.132-0.972; P = .037). In conclusion, the UCP3 -55 C/T SNP was associated with elevated HDL-C levels and a reduced BMI, independent of modifiable factors such as lifestyle. Furthermore, this polymorphism, when expressed in its homozygous form, reduced the prevalence of obesity in Japanese.

BACKGROUND: It has been reported that a common G-->A single nucleotide polymorphism (SNP) at the position -866 of the uncoupling protein-2 promoter (UCP2-866 G/A SNP) modulates UCP2 expression in adipose tissue and pancreatic beta-cell function, and lipid profiles. Reduced low density lipoprotein (LDL) particle size is a significant predictor of the development for coronary artery disease. The purpose of this study was to investigate whether the UCP2-866 G/A SNP was associated with serum LDL particle characteristics in a general Japanese population. MATERIAL/METHODS: In 279 subjects (age 65+/-13 years), body mass index (BMI), percent body fat, blood pressure, and blood biochemical profiles were measured. The UCP2-866 G/A SNP was determined with a fluorescence-based allele-specific DNA primer assay system. LDL particle characteristics were analyzed by high-resolution polyacrylamide gel electrophoresis. RESULTS: The frequency of the -866 A allele was 47.8%. There was no difference in triglyceride, total cholesterol, LDL-cholesterol, HDL-cholesterol, and small dense LDL levels between genotypes. However, subjects with the -866 A/A genotype had significantly lower mean LDL particle size levels (263.5+/-4.9 angstroms) than those with the -866 G/G genotype (264.6+/-4.9 angstroms, P=0.034). Multiple regression analysis revealed that the -866 A/A genotype was a significant variable contributing to the variance in the reduced LDL particle size levels (P=0.012). CONCLUSIONS: The -866 A/A genotype may contribute to reduced LDL particle size levels, a significant risk factor for the development of coronary artery disease.