We investigated the effects of HMG-CoA reductase inhibitors (statins) on the activity and concentration of plasma cholesterol ester transfer protein (CETP) in 30 hypercholesterolemic patients. Patients were divided into three groups according to TaqIB polymorphism of the CETP gene. The activity (158 ± 23% control, mean ± SEM) and concentration (4.1 ± 1.0 mg/l) of plasma CETP were significantly (p < 0.005) higher in the subjects with the B1B1 genotype than B2B2 genotype (106 ± 25% and 2.5 ± 1.1 mg/l, respectively). Plasma CETP activity and concentration levels in the B1B2 group were intermediate between those of the B1B1 and B2B2 groups, and significantly (p < 0.05) low compared with the B1B1 group. Both the activity and concentration of plasma CETP were positively correlated with the LDL-cholesterol concentration (r = 0.608, p < 0.0005 and r = 0.552, p < 0.005, respectively). The administration of statins significantly reduced not only the activity (p < 0.01) but also the concentration (p < 0.05) of plasma CETP in hypercholesterolemic patients. Taken together, we confirmed that statins would be effective in increasing HDL levels in Japanese B1B1 carriers, because of a lower concentration of HDL cholesterol and higher level of plasma CETP compared to the other genotypes. The genetic variation in the CETP gene may be one important factor in designing better treatments.
We tried to investigate whether accumulation of visceral fat assessed by a simple but widely used ultrasonography was associated with common carotid atherosclerosis in women. The subjects were consecutively admitted in-patients whose body mass index was more than 22 kg/m2. Those with cardiorenal or nutritional disorders that would affect blood pressure, and lipid and glucose metabolism were excluded. An ultrasonographic evaluation with a 7.5 MHz linear type B-mode probe was performed by a specialist to determine the intima-media thickness (IMT) of the common carotid artery and maximum thickness of preperitoneal fat (Pmax) at the anterior surface of the liver and the minimum thickness of subcutaneous fat (Smin) of the abdomen. The Pmax/Smin ratio which was termed the abdominal wall fat index (AFI), was then calculated. The subjects were 241 women aged 69 ± 12 (range, 26-104) years. Multiple regression analysis using the IMT as an object variable, adjusted by various risk factors as explanatory variables showed that AFI [odds ratio, 2.995; 95% confidence interval, 1.106-8.109] was a significant independent contributing factor along with known risk factors such as age, systolic blood pressure, total-cholesterol, and HDL-cholesterol. AFI was useful in evaluating disorders of metabolism, circulation and atherosclerosis.
An association between total white blood cell (WBC) count and the risk of coronary heart disease (CHD) is well known. In the present study, we investigated the relationship between WBC counts and other clinical factors including coronary risk factors, and periodontal disease in 1,620 adults. Factors found to have a significant association with the WBC count by multivariate regression analysis were age (negative association), body mass index (negative association), serum triglycerides, high-density lipoprotein cholesterol (negative association), and periodontal levels in the smoking group. Whereas, in the non-smoking group, systolic blood pressure and diastolic blood pressure (negative association) were included and serum triglycerides were excluded. Non-smoking subjects who were free of periodontal disease had a mean WBC count of 4.69 ± 1.0 × 109 cells/l. This value can be used as a reference for WBC counts in healthy subjects. Our data suggest that values significantly higher than this could be an important indicator of inflammation derived from cigarette smoking or periodontal disease. Further study is necessary to determine whether an improvement in the WBC count could prevent the occurrence of CHD.
Cerebral vessels have certain distinct anatomical and developmental characteristics which are well known, but their characteristic genetic expression profile remains as yet only poorly understood. We investigated gene expression in the rat cerebral artery in comparison with the rat descending aorta, two locations which have obviously different anatomical and developmental characteristics. Since the contamination of cerebral small arteries by arachnoid tissue is to a certain extent inevitable, we also performed a gene expression analysis of arachnoid tissue as a background. In an effort to obtain the necessary quality and quantity of total RNA, a novel freeze-fracture apparatus minimizing the time required for the entire procedure from tissue separation to RNA preparation was used. With the material obtained, a group of genes highly expressed in each tissue was detected by oligonucleotide microarray analysis. In the circle of Willis, peptide-19 (PEP-19), connexin-37 (CXN-37), growth arrest-and DNA damage-inducible gene (GADD45), and the putative G protein coupled receptor RA1c, Notch-1, and jagged-1 were predominantly expressed. In arachnoid tissue, bone morphologic protein (BMP)-7, BMP-6, beta defensin-1, neuroendocrine protein 7B2, thiol-specific antioxidant protein, IL-18, beta-chain clathrin-associated protein complex AP-1, and angiopoietin-2 were highly expressed. In the aorta, most of the abundantly expressed genes related to lipid metabolism. By means of oligonucleotide microarray analysis, the distinct gene expression profiles in the circle of Willis arachnoid tissue, and aorta were made evident. From these findings it is reasonable to conclude that a functional interaction exists between the circle of Willis and arachnoid tissue.
Monoclonal antibodies (Mabs) are valuable reagents for the purification, characterization and immunolocalization of proteins. In this study, we raised Mabs against human peroxisome proliferator-activated receptors (PPARs) using baculovirus particles displaying surface glycoprotein gp64-fusion proteins as the immunizing agent. In this system, to display fusion proteins on the viral surface, the amino terminal sequences of human PPARδ and PPARγ2 are inserted in-frame between the signal sequence and the mature domain of the gp64 nucleotide sequence. Mabs were raised by immunization with whole virus without a purification of the target antigens. The Mabs generated by this novel method were shown to recognize not only the gp64-PPARs fusion protein, but also mature, expressed proteins by a wide variety of techniques, including immunohistochemistry, immunoblotting, and electrophoretic mobility shift assays (EMSAs). Transfection of the transfer vector containing a nucleotide sequence encoding less than 30 amino acids along with linearized baculovirus DNA allows for the production of a high affinity antibody against the corresponding mature form. This method is of potential utility in that it allows the production of valuable antibodies without the requirement of a protein purification step.
Tetrahydrocurcumin (THC) is an antioxidative substance which is derived from curcumin by hydrogenation. Curcumin is the main component of turmeric and is responsible for the yellow color of curried foods. First, LDL derived from a normal human volunteer was incubated in the presence of an antioxidant with 10 μM CuSO4 at 37°C for 2 hours. All antioxidants tested (THC, curcumin, probucol, and α-tocopherol) dose-dependently (1-10 μM) inhibited the oxidative modification of LDL. Probucol was the strongest, followed by THC, α-tocopherol, and curcumin. Next, in order to evaluate the antioxidative activity of THC in vivo, we fed rabbits diets containing 1% cholesterol with or without 0.5% THC and examined their effects on oxidative stress and atherosclerosis. Animals were divided into two groups: the control group rabbits (n = 12) were fed a normal chow diet and the experimental group (n = 12) was fed a diet containing 0.5% THC for one week. Then, 1% cholesterol was added to the diets and the animals were allowed to feed further for either 6 (n = 4 for each group) or 12 weeks (n = 8 for each group). Although serum cholesterol levels rapidly increased after starting the high cholesterol diet, no difference was observed between the control and THC groups. TBARS formation in the absence of added copper ion was inhibited in the LDL separated from THC-treated animals compared with that from control animals. THC treatment tended to inhibit the area covered with atherosclerotic lesions compared with the control, although this was not significant (28.8 ± 17.5% vs. 40.0 ± 23.7%, p = 0.2). Formation of Nε-(hexanoyl) lysine, 4-hydroxynonenal and dityrosine in liver and kidney also had a tendency to be inhibited by THC treatment. Although free THC was not detected in serum and liver, THC was detected in samples treated with β-glucuronidase and sulfatase, suggesting that THC is present as a conjugate with glucuronic acid or sulfate. In conclusion, the present results suggest that curcuminoids, particularly THC, which are contained in turmeric, may be useful as a functional food factor.
The purpose of Holicos-PAT was to investigate the efficacy of serum lipid lowering by pravastatin against coronary heart disease (CHD) and cerebrovascular disease (CVD) in the Japanese population. Hypercholesterolemic men and women (n = 2,232), aged 40-70 years, were followed up for 5 years, while they were receiving pravastatin (group P, n = 1,422) or only diet therapy (group C, n = 810). The primary endpoint was CHD (a composite of onset or worsening of angina pectoris, performing CABG or PTCA, non-fatal myocardial infarction, death from CHD including heart death or sudden death). The secondary endpoints were comprised of CVD, total mortality, variation of serum lipid and apoprotein levels, and a relationship between the LDL-C level and occurrence of CHD. For several reasons (proving to meet the exclusion criteria after registration, etc.), 1,290 cases of group P and 749 cases of group C were used as subjects for the primary analysis. The mean follow-up period was 4.5 years in group P and 4.2 years in group C for events of CHD. The mean LDL-C level (SD) in group P was 176 (29) mg/dl and decreased to 134 (29) mg/dl one year later. This effect continued during the follow-up period. CHD events occurred in 9.2/1000 patient-years for men and 2.4/1000 patient-years for women without a history of CHD. CHD events occurred in 55.3/1000 patient-years for men and 23.6/1000 patient-years for women with a history of CHD, which was 6 times higher in men and 10 times higher in women than in those without a history of CHD, respectively. The adjusted relative risk ratio of group P to group C for CHD events was 0.74 (95%Cl: 0.47-1.19). In the patients with a history of CHD, the ratio was 0.55 (95%Cl: 0.30-1.00). The effect was apparent in the patients with a history of CHD. The incidence of myocardial infarction in Japanese patients with hypercholesterolemia living in the Hokuriku district was apparently lower, than the worldwide incidence, indicative that pravastatin may have a tendency to inhibit the occurrence of events of arteriosclerotic disease.