Aim: Familial hypercholesterolemia (FH) patients are at particular risk for premature coronary artery disease (CAD) caused by high levels of low density lipoprotein (LDL). Administration of statins enabled us to reduce LDL-C levels in heterozygous FH patients. To evaluate the impact of statins on the clinical fate of heterozygous FH, a retrospective study was performed.
Methods: We analyzed the clinical influence of statins on age at the first clinical onset of CAD in 329 consecutive FH patients referred to the lipid clinic of the National Cardiovascular Center. Among 329 heterozygous FH patients, the onset of CAD was identified in 101.
Results: The age at onset of CAD was 58.8±12.5 years in the 25 patients on statins at onset, significantly higher than that in the 76 patients not on statins (47.6±10.5 years) (p <0.001). The average age at CAD onset was significantly higher after widespread use of statins (54.2±13.2 years in 48 patients, Group 1) compared to before October 1989 when statins were approved in Japan (46.9±9.6 years in 53 patients; Group 2, p=0.002). A significant difference was seen between Groups 1 and 2 in the variables, including sex, prevalence of smoking habit, LDL-C, and the use of statins, aspirin and probucol. After adjusting for these variables, only statin use was independently associated with the difference in age at CAD onset by multivariable analysis.
Conclusion: Statins have improved the clinical course of patients with heterozygous FH.

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Aim: This immunohistochemical investigation was to analyze the relationship between platelet activation/aggregation, inflammatory cell infiltration, the differentiation state of neointimal smooth muscle cells (SMCs), expression of platelet-derived growth factor (PDGF), and endothelial cell regeneration at sites of bare-metal stents (BMS) in patients with acute coronary syndrome (ACS).
Methods: Sixteen coronary arteries after stenting were obtained at autopsy from ACS patients. Serial frozen sections were stained with antibodies against SMCs (1A4, HHF-35, CGA-7), macrophages, neutrophils, endothelial cells, GP IIb/IIIa, P-selectin, PDGF-B, and PDGF-β receptor.
Results: Up to 12 days after BMS, the stent sites contained P-selectin-positive activated platelets with neutrophil infiltration. From 24 to 55 days after BMS, parts of the platelet thrombi were still positive for P-selectin, and infiltration of neutrophils and macrophages was also found. Neointimal SMCs at these stages stained positive with 1A4 but negative with CGA-7, and PDGF-B and PDGF-β receptor were expressed in macrophages and SMCs. At sites from 3 months onward, platelet thrombi and neutrophil infiltration were not detected, and the neointima contained increased numbers of highly differentiated SMCs with CGA-7 positivity. The P-selectin-positive area was positively correlated with the neutrophil count and macrophage-positive area (neutrophils, r=0.86, p<0.0005; macrophage, r=0.66, p<0.05). In contrast, the P-selectin-positive area was negatively correlated with the HHF-35-positive area and CGA-7-positive area (HHF-35, r=-0.90, p<0.0001; CGA-7, r=-0.82, p<0.005).
Conclusion: These observations suggest that P-selectin-positive platelet thrombi in the neointima are positively associated with the inflammatory cell infltration and reversely associated with the phenotypic redifferentiation of neointimal SMCs after BMS in ACS patients.

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Aim: The present study examined the association between the radial augmentation index (AI), a marker of arterial wave reflection, and the MEGA risk prediction score (MEGA score), an indicator of coronary heart disease (CHD) risk, in middle-aged men with mild to moderate hypercholesterolemia.
Methods: Radial AI was measured during a company health examination in 266 men (age: 47±5 years) with total cholesterol levels ranging 220-270 mg/dL who were not taking antihypertensive, lipid-lowering, or antidiabetic agents. The MEGA score was calculated based on sex, age, low- and high-density lipoprotein cholesterol, blood pressure, glucose level, and smoking status. The higher MEGA score indicates increased CHD risk. A MEGA score ≥ 22 corresponds to a 5-year CHD risk ≥ 2.5% and we defined a MEGA score ≥ 22 as a high estimated CHD risk.
Results: The mean AI was 74.4±12.6%. A high estimated CHD risk was seen in 32 subjects (12.0%). After adjusting for height and heart rate, the AI was higher in subjects with a high estimated CHD risk (81.5±10.6%) than in those without (73.4±10.4%, p<0.001). The odds ratio for high estimated CHD risk in the highest tertile of AI was 8.14 (p=0.002) in comparison to the lowest tertile, after adjusting for multiple potential confounders which did not constitute the MEGA score.
Conclusion: The radial AI was positively associated with the estimated risk of CHD. These results suggest the usefulness of radial AI as a risk marker for future onset of CHD in middle-aged men with mild to moderate hypercholesterolemia.

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Aim: The aim of this study was to clarify the relationship between alcohol intake and metabolic syn-drome in Japanese men and women.
Methods: Japanese female subjects (n=11,187) were divided into non-, light (<22 g ethanol/day) and heavy (≥ 22 ethanol/day) drinkers, and Japanese male subjects (n=19,398) were divided into non-, light (<22 g ethanol/day), heavy (≥ 22 and <44 g ethanol/day) and very heavy (≥ 44 g ethanol/day) drinkers. The mean level of each variable and the prevalence of each risk factor and metabolic syndrome were compared among the groups.
Results: In men and women, blood pressure and HDL cholesterol tended to be higher, and hemoglobin A1c tended to be lower with increased alcohol intake. Waist circumference showed U- and V-shaped relationships, and log-converted triglyceride showed J- and V-shaped relationships with alcohol intake in men and women, respectively. The prevalence of metabolic syndrome was lowest in light drinkers in men and women and was significantly higher in very heavy drinkers than in non-drinkers in men. In men, the odds ratio vs. non-drinkers for metabolic syndrome was significantly low in light drinkers, while the odds ratio was significantly high in very heavy drinkers. In women, a significantly low odds ratio vs. non-drinkers for metabolic syndrome was obtained in light drinkers.
Conclusion: Light drinking is associated with a lower risk of metabolic syndrome in Japanese men and women, while very heavy drinking is thought to increase the risk of metabolic syndrome in Japanese men.

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Aim: The clinical relevance of the suggested pleiotropic effects of hydroxymethylglutaryl coenzyme A reductase inhibitors (statins) is controversial. Aggressive statins effectively reduce lipid levels, but whether their other effects are more powerful than those of regular statins is unknown.
Methods: We enrolled 32 patients (mean age, 65 y; male, 23) who had undergone coronary revascularization over 6 months previously and whose serum LDL cholesterol levels persisted at >100 mg/dL, regardless of pravastatin (10 mg/day). Before and 1 and 6 months after switching to atorvastatin (10 mg/day), we evaluated lipid profiles, including RLP-C (remnant-like particle cholesterol), high sensitive CRP (hsCRP), soluble CD40 ligand (sCD40L), TBARS (thiobarbituric acid reactive substances), and endothelial function determined from flow-mediated dilation (FMD) of the brachial artery.
Results: One month on atorvastatin lowered LDL cholesterol by 24% (131 to 100 mg/dL, p<0.001). In addition, RLP-C, sCD40L and hsCRP significantly decreased, whereas FMD did not change. After 6 months of this therapy, FMD significantly improved compared to baseline values (5.1 vs 3.6%, p=0.04). Changes in FMD and in total and RLP cholesterol significantly correlated. Moreover, FMD was remarkably improved in patients who achieved target LDL levels (<100 mg/dL).
Conclusions: Switching from a regular to an aggressive statin can improve endothelial function at 6 months in patients with previous coronary artery disease. This effect is suggested to be mainly due to the lipid-lowering effect. Achievement and maintenance of the target LDL level by switching statins is beneficial in the clinical setting.

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Aim: Research suggests that sex may condition atherosclerosis development against different genetic backgrounds. This study addresses the hypothesis that this effect would be exerted by changes in the different apolipoproteins present in high-density lipoproteins.
Methods: ApoE-deficient mice of both sexes with Ola 129 and C57BL/6J genetic backgrounds were fed a chow diet for 14 weeks. At the end of the dietary intervention, the development of atherosclerotic lesions, apolipoproteins, lipid metabolism, inflammation and paraoxonase were assessed.
Results: Differences between atherosclerotic lesions in Ola 129 and C57BL/6J strains of apoE-deficient mice were sex-dependent and were only statistically significant in females. Plasma levels of HDL cholesterol and apolipoproteins related to these lipoparticles, such as apoA-I, apoA-II, apoA-IV, apoA-V and apoJ, were significantly different between these two strains and there were sex-related differences in some of these apolipoproteins. Hepatic steatosis was also related to the strain and was independent of sex. In females, changes in HDL cholesterol and apolipoproteins A-I and A-II were important determinants of atherosclerosis, while this was not the case in males.
Conclusions: Our results demonstrate that atherosclerosis-related differences between Ola129 and C57BL/6J genetic backgrounds in apoE-deficient mice are sex-dependent and that this finding is explained by the differences in HDL cholesterol and its apolipoprotein components, mainly apoA-I and A-II. Overall, our findings highlight the importance of taking sex into account in the analysis of atherosclerosis and lipid metabolism in animal models.

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Aim: Hypercholesterolemic patients with inflammation are at high risk for cardiovascular events. Statins exert anti-inflammatory action independent of their cholesterol-lowering action. This study sought to clarify whether statin therapy reduces inflammatory markers in hypercholesterolemic patients and to determine factors that predict the reduction in these markers.
Methods: Fasting concentrations of lipoproteins and inflammatory markers were measured in 54 hypercholesterolemic patients, and age- and gender-matched healthy volunteers. Carotid atherosclerosis was determined by ultrasonography. Blood samples were also analyzed in hypercholesterolemic patients after 4 weeks of statin therapy.
Results: The high-sensitivity C-reactive protein (hs-CRP) and serum amyloid A (SAA) protein concentrations did not differ between the two groups. Statin therapy reduced the median hs-CRP and SAA concentrations in hypercholesterolemic patients from 0.75 to 0.60 mg/L (p=0.05), and from 3.95 to 3.20μg/mL (p=0.20), respectively. These reductions were significant for both markers, but only in subgroups with high baseline concentrations. Statins exhibited different results for hs-CRP and SAA in the presence of carotid atherosclerosis.
Conclusions: Statin therapy reduces inflammatory markers in hypercholesterolemic patients, and this anti-inflammatory action is limited to patients whose inflammatory markers are elevated at baseline.

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Aim: Platelets plays a central role in hemostatic processes and consequently are similarly involved in pathological processes, such as arterial thrombosis and atherosclerosis. Herein we described the synthesis, antiplatelet profile and structure-activity relationship (SAR) of a new series of N'-substitutedphenylmethylene-1H-pyrazolo[3,4-b]pyridine-carbohydrazide derivatives (3a-3k).
Methods: These compounds were synthesized in good yield and tested in platelet aggregation assays using collagen, ADP and arachidonic acid as agonists. We also performed a SAR studies using SPARTAN' 08 program, in silico ADMET screening and the Lipinski “ rule of five ” using Osiris Property Explorer and molinspiration on-line programs.
Results: Interestingly, the new compounds were active against collagen and arachidonic acid (AA) with the two most actives compounds (3a and 3c - IC₅₀=61 μM and 68 μM respectively) almost 5-fold more potent than aspirin (IC₅₀=300 μM). These derivatives showed low theoretical toxicity risks in in silico ADMET screening and fulfilled the Lipinski rule of five, suggesting good oral biodisponibility.
Conclusion: This work showed carbohydrazide group as potential for designing new antiplatelets. On that purpose, 3a and 3c may act as prototypes to generate more efficient and safe molecules for treating thrombotic diseases.

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Aim: Acyl-coenzyme A: cholesterol acyltransferase1 (ACAT1) is an endoplasmic reticulum (ER)-resident enzyme that catalyzes the conversion of cholesterol into cholesteryl esters. We previously showed that cholesterolloaded macrophages produce numerous ER-derived vesicles with elevated ACAT1 enzyme activity; some of these vesicles were shown to be closely associated with Golgi-related organelle(s). The aim of this study was to investigate the translocation of ACAT1 vesicle in cholesterol-loaded macrophages.
Methods: To demonstrate association of ACAT1 with late endosomes/lysosomes (LE/LS), primary human macrophages with or without cholesterol-loading was subjected to confocal microscopy, immunoelectron microscopy, subcellular fractionation, and immunoadsorption assay. Furthermore, cholesterol esterification assay was also carried out to investigate function of ACAT1 associated LE/LS.
Results: Confocal fluorescence microscopy revealed that no significant ACAT1 signal was associated with the signal for LAMP2, a marker protein for LE/LS, in cholesterol non-loaded macrophages; however, approximately 20% of the total ACAT1 signals colocalized with the LAMP2 signal in cholesterol-loaded macrophages. ACAT1-positive membranes isolated by immunoadsorption using ACAT1-specific antibody contained LAMP2, demonstrating the association of ACAT1 and LE/LS. In addition, in macrophages phagocytosing latex beads, the close association of ACAT1 with LE/LS can be demonstrated in phagosomes isolated from cholesterol-loaded macrophages, not from non-loaded macrophages. Furthermore, cholesterol-loaded macrophages re-esterified aggregated LDL-derived cholesteryl ester even in the presence of U18666A, a reagent known to block egression of cholesterol from LE/LS.
Conclusion: Our results indicated that cholesterol-loaded human macrophages produce LE/LS in close association with ACAT1, and may promote efficient esterification of modified LDL-derived free cholesterol on LE/LS.

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Aim: Peripheral arterial disease (PAD) is associated with cerebrovascular disease, ischemic heart disease, and other cardiovascular disease. We investigated the prevalence of and factors related to PAD to clarify the relationship between PAD and carotid atherosclerosis in a cross-sectional populationbased study.
Methods: The study included 2,402 (900 males and 1,502 females; mean±SD=64.9±10.9 years) of 3,862 residents of two Japanese rural areas who reported for a free health examination in 2005 or 2006. An ankle brachial index value ≤0.9 was considered to be PAD. The carotid artery intima-media thickness (CA-IMT) was measured by carotid ultrasound.
Results: The prevalence of PAD was 1.71% (n=41) of all participants. The risk factors independently associated with a significantly higher risk of PAD, identified by multivariate analysis, are as follows: For males, age, dyslipidemia, and CA-IMT, and for females, age, waist circumference, and dyslipidemia.
Conclusion: The prevalence of PAD in Japan was confirmed to be lower than that of similar studies performed in other countries. PAD was strongly correlated with age and dyslipidemia in both sexes, carotid atherosclerosis in males, and abdominal fat in females.

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Aims: Whether a low ankle-brachial index can improve the prediction of all-cause and cardiovascular mortality on top of conventional risk factors remains unclear among patients with ischemic heart disease. The present study aimed to assess the association between the ankle-brachial index and mortality in Chinese patients.
Methods: This was an observational prospective study and 1,800 Chinese patients aged ≥35 years were followed-up from 2004 to 2007-2008.
Results: There were 280 deaths, of which 165 were attributable to cardiovascular disease. Compared with patients with an ankle-brachial index ≥1.1, the risk of mortality increased linearly in lower ankle-brachial index categories: patients with an ankle-brachial index of 0.9 to 1.1, 0.7 to 0.9, <0.7 had hazard ratios of 1.60, 2.07, and 3.08 for all-cause mortality and 1.89, 2.33, and 4.09 for cardiovascular mortality (p for trend <0.001), respectively. Addition of the ankle-brachial index significantly (p<0.001) increased the predictive value of the model for 3-year deaths compared with a model including risk factors alone. Comparison of areas under receiver operator characteristics curves confirmed that a model including the ankle-brachial index discriminated better than without.
Conclusion: There was an inverse association between the ankle-brachial index and mortality. Addition of the ankle-brachial index significantly improved the prediction of 3-year mortality over and above that of conventional risk factors. We recommend that the ankle-brachial index be incorporated into prognostic assessment for patients with ischemic heart disease.

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A 67-year-old man was admitted to our institution with sudden and pessistant chest pain for 3 days. Coronary angiography showed massive thrombotic occlusion of the right coronary artery. The patient received intracoronary thrombolysis with alteplase (recombinant tissue-type plasminogen activator, rt-PA). On repeated angiography, there was marked resolution of intracoronary thrombus. After percutaneous coronary intervention with stent implantation, the final result was complete revascularization of the right coronary artery (TIMI grade 3 distal flow). This case demonstrates that intracoronary rt-PA can result in local thrombus reduction in patients undergoing PCI, especially with a large thrombus burden.

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We describe a diabetic patient successfully treated for an acute mycotic aortic arch pseudoaneurysm with primary aldosteronism. The patient first complained of severe pain in the left upper extremity and left back with high C reactive protein (CRP) and high-grade fever. It was suspected that acute aortic dissection had developed in association with mycotic pseudoaneurysm of the aortic arch because of chest X-ray findings of enlargement of the aortic arch. Computed tomography (CT) of the aortic arch revealed an aortic aneurysm protruding in the superior direction. Staphylococcus aureus was detected in blood culture, suggesting a mycotic aortic aneurysm, and artificial blood vessel replacement of the aortic arch was performed. Intraoperative findings suggested aortic pseudoaneurysm, which consisted of mediastinal rupture of the aorta at the distal arch. Our patient had a 2-year history of type 2 diabetes mellitus and poor blood sugar control, even with twice-daily injection of insulin. Blood pressure was not always well controlled because of primary aldosteronism. Thus, it was speculated that hyperaldosteronism, as well as diabetes-associated atherosclerosis, had persisted for a long time. No reports have described mycotic pseudoaneurysm in the aortic arch in a diabetic patient associated with primary aldosteronism. It is necessary to note that serious vascular complications are possible if aldosteronism is left untreated or is treated insufficiently as essential hypertension.

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