Aim: Small, dense low density lipoprotein (sLDL) is known as an atherogenic lipoprotein and is often associated with metabolic syndrome (MS). A high frequency of sLDL is found in hypertriglyceridemic subjects. Also, fatty liver (FL) is often associated with MS; therefore, we studied whether the association of FL increases sLDL- cholesterol (C ) in subjects with MS.
Methods: In total, 207 patients were enrolled in this study and FL was estimated by echogram. The presence of MS was diagnosed according to the Japanese Guidelines for the Definition of Metabolic Syndrome.
Results: sLDL-C and sLDL-C/LDL-C in the MS group were higher than in the non-MS group. Also, sLDL-C and sLDL-C/LDL-C in the FL group were higher than in the non-FL group. The simple correlation coefficient (r) between plasma triglyceride and sLDL-C or sLDL-C/LDL-C in all subjects was 0.36 and 0.51. In the MS group, r values were 0.32 and 0.52 while, in the non-FL group, r was 0.32 and 0.38, respectively. Two-way ANOVA revealed that FL was a powerful determinant of plasma sLDL-C and sLDL-C/LDL-C, but MS was not. When we divided all subjects into four groups, i.e., MS(-)FL(-), MS(-)FL(+), MS(+)FL(-) and MS(+)FL(+), sLDL-C/LDL-C of MS(+)FL(+) was significantly higher than all other groups.
Conclusion: Association of MS and FL significantly increased sLDL-C and sLDL-C/LDL-C. The significant relationship between sLDL-C/LDL-C and plasma triglyceride in the FL group indicates that FL may produce triglyceriderich VLDL, a precurser of sLDL, thereby contributing to the appearance of sLDL particles in the plasma of MS patients with FL.

View full abstract

Aim: Pitavastatin significantly improved lipid profiles and reduced serum high-sensitivity C-reactive protein (hs-CRP) levels in a multi-center and prospective study. The aim of this study was to explore the effect of pitavastatin on serum levels of another inflammatory biomarker, interleukin-18 (IL-18), in a sub-analysis of the previous multi-center prospective study.
Methods: The subjects were 83 patients derived from the KISHIMEN study. Pitavastatin (1-2 mg/day) was administered for 12 months. Serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), remnant-like particle cholesterol (RLP-C), triglycerides (TG), IL-18, and high sensitivity C-reactive protein (hs-CRP) levels were measured.
Results: TC, LDL-C, and RLP-C levels were significantly reduced by 18.3%, 30.1%, and 21.0% (mean values) at 12 months after pitavastatin administration. TG levels were decreased by 9.8% in subjects whose basal TG levels were above 150 mg/dL. HDL-C levels were significantly increased at 6 months (11.9%). Pitavastatin did not significantly alter IL-18 levels in overall subjects, but reduced IL-18 levels in the highest quartile by 24.5% (median value) at 12 months. Pitavastatin significantly reduced hs-CRP levels by 28.6% in overall subjects and by 62.4% in the highest quartile at 12 months. There was a significant correlation between IL-18 and hs-CRP at baseline after both values were transformed into logarithms (Pearson's correlation coefficient, r=0.259, p=0.0181); however, percent changes in these levels were not significantly correlated.
Conclusion: Pitavastatin significantly improves lipid profiles, and reduces enhanced inflammation monitored by IL-18, as well as by hs-CRP, in hypercholesterolemic subjects.

View full abstract

Aim: We hypothesized that excessive suppression of platelet function due to antiplatelet therapy can increase the incidence of bleeding complications. The aim of the present study was to find whether we could predict bleeding events by measuring platelet function.
Methods: We enrolled 743 subjects whose platelet function was measured using a whole blood aggregometer based on a screen filtration pressure method. Of these subjects, 551 (74.2%) were treated with some type of antiplatelet agent. The endpoints were bleeding or ischemic events requiring hospitalization or extension of hospital stay. We prospectively compared the platelet function of subjects with and without bleeding or ischemic events.
Results: During 556±207 days of follow-up, 52 (7.0%) bleeding events and 20 (2.7%) ischemic events were observed. Kaplan-Meier analysis using the log-rank test revealed that an aggregation rate of < 20% induced by 8 μM adenosine diphosphate (ADP) was significantly associated with a greater number of bleeding events (11.9% vs. 5.2%; p=0.0007). Cox proportional hazards model showed that age > 75 years (hazard ratio [HR], 1.78; 95% confidence interval [CI], 1.03-3.10; p=0.039), estimated glomerular filtration rate < 60 ml/min/1.73m² (HR, 1.82; 95% CI, 1.06-3.18; p=0.031) and aggregation rate < 20% induced by 8 μM ADP (HR, 2.18; 95% CI, 1.24-3.80; p=0.0071) were independent predictors of bleeding events.
Conclusions: Low platelet function demonstrated using a whole blood aggregometer was an independent predictor of bleeding complications.

View full abstract

Aim: Hyperplasia suppressor gene/mitofusion-2 (HSG/Mfn2) is a hyperplasia suppressor gene and an essential component of mitochondrial fusion machinery; however, the association between the single nucleotide polymorphism (SNP) of HSG/Mfn2 and hypertension is unclear.
Methods: In this study, 542 normotensive subjects (NT group) and 539 hypertensive patients (EH group) were screened for an association study between HSG/Mfn2 and hypertension.
Results: The results showed that the genotype distribution and allelic frequency of rs873457, rs2336384, rs1474868, rs4846085 and rs2236055 were significantly different (p lt; 0.05 for all) between EH and NT groups, although those of rs4240897 and rs873458 were not. When comparing the dominant model, significant differences still existed (p lt; 0.05 for all). The allelic frequency of rs4240897 was also slightly different between EH and NT groups (P=0.047). When subgrouped by sex, the genotype distribution and allelic frequency of all the SNPs (except rs873458) were significantly different in male (p lt; 0.05 for all) but not in female groups. For all the SNPs, only the allelic frequency of rs4240897 was obviously different in female NT and EH groups (p lt; 0.01). Logistic regression showed that body mass index and rs873457 were closely associated with BP after adjusting for age. The frequency of the C-G-A-A-A-C-C haplotype was significantly higher in essential hypertensive patients versus control individuals, both in the entire population, in male or female groups (p lt; 0.01 for all). As for other haplotypes, most were only significantly different in the entire population and male subjects.
Conclusion: The genetic variations of HSG/Mfn2 may be associated with hypertension in male Chinese.

View full abstract

Aims: Hypertrophied hearts at autopsy often display excessive coronary artery atherosclerosis, but the histopathology of coronary arteries in hearts with and without cardiomegaly has rarely been compared.
Methods: In this study, forensic autopsies provided hearts with unexplained enlargement plus comparison specimens. Right coronary artery was opened longitudinally and flattened for formalin fixation and H&E-stained paraffin sections were cut perpendicular to the endothelial surface. The micro-scopically observed presence or absence of a necrotic atheroma in the specimen was recorded. At multiple sites far removed from any form of atherosclerosis, measurements were taken of intimal thickness, numbers of smooth muscle cells (SMC) and their ratio, the thickness per SMC, averaged over the entire nonatheromatous arterial length. When the mean thickness per SMC exceeded a certain cutoff point, the artery was declared likely to contain a necrotic atheroma.
Results: The prevalence of specimens with necrotic atheromas increased stepwise with increasing heart weight, equally with fatal or with incidental cardiomegaly, and equally with hypertension- or obesity-related hypertrophy, rejecting further inclusion of appreciable age, race, or gender effects. The prevalence of specimens with thickness per SMC exceeding the cutoff point was almost always nearly identical to the prevalence of observed necrotic atheroma, showing the two variables to be tightly linked to each other with quantitative consistency across group comparisons of every form.
Conclusions: In summary, cardiomegaly, irrespective of the specific cause, seems to accelerate the risk of atheromas, and to do so by first altering the arterial architecture, especially by increasing intimal thickness per SMC.

View full abstract

Aim: Some randomized studies have shown a delay of up to a few years in the statin-related survival advantage, whereas others have demonstrated an early survival benefit for some patients. We examined the short-term effects of statins in patients with acute coronary syndrome (ACS), stratified according to baseline LDL-C.
Methods: Patients with ACS (n=180) were randomized to receive 6 months of atorvastatin (20 mg) in the Extended-ESTABLISH trial. Six months after ACS onset, all patients were treated with statins to achieve an LDL-C value of < 100 mg/dL. Patient outcomes were analyzed with respect to LDL-C at the time of ACS onset: high baseline (≥100 mg/dL, n=124) or low baseline (< 100 mg/dL, n=56) LDL-C.
Results: The cumulative incidence rates of major adverse cardiac and cerebrovascular events (MACCE) did not significantly differ between the early-statin and control groups in the high baseline groups at 6 months (p=0.158), whereas a significant benefit of early intensive statins appeared 1 year (p=0.034) later. In contrast, we found no significant short-term benefits of statins after either 6 months or 1 year in the low baseline group. Multivariate analysis showed that early intensive atorvastatin therapy was associated with a lower risk of MACCE at 1 year in the high baseline group (OR, 0.25; 95% CI, 0.05 to 0.83; p=0.035).
Conclusions: The effects of 6 months of intensive lipid-lowering therapy appear after 1 year in patients with ACS and baseline LDL-C ≥100 mg/dL.

View full abstract

Aim: The cardio-ankle vascular stiffness index (CAVI) is a new parameter that reflects the stiffness of the aorta, femoral artery and tibial artery as a whole. One of its conspicuous features is that CAVI is independent of blood pressure at measuring time, theoretically. But, it has not been experimentally proved yet. For confirmation, pharmacological studies were performed comparing with brachial-ankle pulse wave velocity (baPWV).
Methods: Used drugs were a β1-adrenoceptor blocker, metoprorol and an α1- adrenoceptor blocker doxazosin. Both were administered to 12 healthy volunteer men. CAVI and baPWV were measured every one hour for 6 hours using VaSera.
Results: When metoprolol (80 mg) was administered to 12 healthy volunteer men, systolic blood pressure decreased from 131.4±4.5 to118.3±4.1 mmHg (mean±SE) (p< 0.05) at the 3rd hour, and diastolic blood pressure decreased from 85.3±4.0 to 75.3±3.0mmHg (p< 0.05). baP-WV decreased from 13.93±0.46 to 12.46±0.49 m/sec (p< 0.05), significantly, but CAVI did not change (8.16±0.29 to 8.24±0.27) (p=0.449). ΔbaPWV at each time was significantly correlated with both Δsystolic and Δdiastolic blood pressures, but ΔCAVI was not correlated with either Δblood pressure. When doxazosin (4 mg) was administered to the same men, systolic blood pressure decreased from 130.2±4.6 to117.2±4.8 mmHg (p< 0.05) at the 3rd hour. Diastolic blood pressure also decreased from 85.1±4.1 to 74.2±3.9 mmHg (p< 0.05). baPWV decreased from 13.98±0.68 to 12.25±0.53 m/sec (p< 0.05), significantly. CAVI also decreased from 8.15±0.28 to 7.18±0.37 (p< 0.05), significantly.
Conclusion: These results suggested that CAVI was not affected by blood pressure at the measuring time directly, but affected by the changes of contractility of smooth muscle cells.

View full abstract

Aim: The current study aimed to address the inconsistencies in association studies, specifically with reference to methylene tetrahydrofolate reductase (MTHFR) C677T polymorphism in the light of gene-gene and gene-nutrient interactions.
Methods: A case-control study was conducted to analyze four genetic polymorphisms i.e. thymidylate synthase (TYMS) 5'-UTR 28 bp tandem repeat, MTHFR C677T, methyltetrahydrofolate homocysteine methyltransferase (MTR) A2756G, methyltetrahydrofolate homocysteine methyltransferase reductase (MTRR) A66G using PCR-AFLP and PCR-RFLP methods; plasma folate and B12 using AxSYM kits; plasma homocysteine by reverse phase HPLC and nitric oxide using Griess reaction. Fisher's exact test, logistic regression analysis and multifactor dimensionality reduction analysis were used for statistical analysis of genetic parameters. Student's t-test was used for biochemical parameters.
Results: MTHFR C677T and MTRR A66G were found to increase the risk for CAD by 1.61-fold (95% CI: 1.04-2.50) and 1.92-fold (95% CI: 1.29-2.87) whereas TYMS 2R allele was found to reduce the risk for CAD (OR: 0.66, 95% CI: 0.49-0.88) by counteracting MTHFR and MTRR variant alleles. Significant gene-gene interactions were observed among TYMS/MTRR (P < 0.0001), MTR/TYMS/MTRR (P < 0.0001), and MTHFR/MTR/TYMS/MTRR (P < 0.0001). MTHFR was found to increase the risk (OR: 2.36, 95% CI: 1.28-4.37) only in the absence of the TYMS 2R allele, with marked impairment of the remethylation process (P=0.007). This impairment was predominant when the dietary folate was in the lowest tertile. In subjects with dietary folate intake in the highest tertile, no such impairment was observed.
Conclusion: Dietary folate status and TYMS 5'-UTR 28bp tandem repeat polymorphism are important effect modifiers of CAD risk associated with genetic variants in remethylating genes.

View full abstract

Aim: Microalbuminuria (MAU) and heart rate are established predictors of an adverse cardiovascular outcome. Recently, heart rate was described as an independent predictor of MAU in hypertensive patients, raising the question of a causal link.
Methods: In post-hoc analysis of the PROactive trial we examined the association of the baseline heart rate and MAU in diabetic patients with cardiovascular disease (n=5,110, mean age 62±8, 66% male) using logistic regression. Cox regression analysis was used to examine the independent impact of heart rate and MAU on the composite endpoint of all-cause mortality, myocardial infarction and stroke.
Results: Baseline heart rate was not associated with a significantly increased risk for MAU at baseline (OR 1.01 per 10 bpm, 95% CI 0.97-1.06, p=0.48) or MAU at the final visit (OR per 10 bpm 1.04, 95% CI 0.98-1.11, p=0.20). Similar results were observed in subgroups of patients with hypertensive blood pressure at baseline (OR 0.98 per 10 bpm, 95% CI 0.93-1.03, p=0.42) or patients with a history of hypertension (OR 1.02 per 10 bpm, 95% CI 0.98-1.07, p=0.31), respectively. Stratification by use of an ACE inhibitor/AT1-receptor blocker did also not change the results. In multivariate analysis, both heart rate and MAU were significantly predictive of a cardiovascular outcome.
Conclusion: There was no evidence of an association between heart rate and MAU in diabetic patients with cardiovascular disease, independently of whether hypertension was present or not, but both markers were independently predictive of a cardiovascular outcome. These results do not support a causal link between heart rate and MAU.

View full abstract

Aim: To determine whether culturable bacterial strains are present in human atheromatous tissue and to investigate their properties using culture, quantitative PCR, metagenomic screening, genomic and biochemical methods.
Methods: We analyzed femoral atherosclerotic plaque and five pairs of diseased and healthy arterial tissue for the presence of culturable bacteria using cell cultures and genomic analysis.
Results: Gram negative aerobic bacilli were cultivated from the plaque tissue. Ribosomal 16S DNA amplification and sequencing identified the isolates as Enterobacter hormaechei. The isolate was resistant to ampicillin, cefazolin, and erythromycin. A circular 10kb plasmid was isolated from the strain. Antibiotic protection assays of the isolate demonstrated invasive ability in a human monocytic cell line. To extend the study, five matched pairs of diseased and healthy aortic tissue were analyzed via quantitative PCR. Eubacterial 16S rDNA was detected in all specimens, however, E. hormaechei DNA was detected in surprisingly high numbers in two of the diseased tissues only.
Conclusions: While it is well documented that inflammation is an important risk factor for vascular pathophysiology, the association of bacteria with atherosclerosis has not been clearly established, in large part due to the inability to isolate live bacteria from atheromatous tissue. This is the first study providing direct evidence of Enterobacter spp. associated with atheromatous tissues. The data suggest that chronic infection with bacteria may be an under-reported etiologic factor in vascular pathogenesis. Importantly, characterization of the clinical isolate supports a model of atherogenesis where systemic dissemination of bacteria to atherosclerotic sites may occur via internalization in phagocytic cells.

View full abstract