Effects of fenofibrate therapy on circulating adipocytokines in patients with primary hypertriglyceridemia

BackgroundWe investigated effects of fenofibrate therapy on endothelial dysfunction and adipocytokine profiles.MethodsA randomized, single-blind, placebo-controlled, cross-over study was conducted in 53 patients with primary hypertriglyceridemia. We administered placebo or fenofibrate 160 mg daily for 8 weeks.ResultsWhen compared with placebo, fenofibrate therapy substantially lowered plasma levels of TNF-α by 6 ± 3% (P = 0.014) and hsCRP from 1.10 to 0.90 mg/l (P = 0.004). When compared with placebo, fenofibrate therapy increased plasma levels of adiponectin by 17 ± 4% (P = 0.001), insulin sensitivity by 4 ± 1% (as assessed by QUICKI, P = 0.009), and decreased plasma levels of leptin and resistin by 4 ± 7% (P = 0.022) and 10 ± 3% (P = 0.001), respectively. There were correlations between percent changes in QUICKI and percent changes in adiponectin levels (r = 0.279, P = 0.043) or leptin (r = ?0.280, P = 0.042).ConclusionsFenofibrate therapy significantly reduced pro-inflammatory biomarkers and improved adipocytokines levels and insulin sensitivity in hypertriglyceridemic patients.     

Impact of type 2 diabetes mellitus on diffuse inflammatory activation of de novo atheromatous lesions: Implications for systemic inflammation

AimsLocal coronary and systemic inflammation is pronounced in patients with diabetes mellitus (DM). Intracoronary thermography detects local inflammation and C-reactive protein (CRP) is a marker of systemic inflammation. We investigated whether or not, in patients with DM, thermal heterogeneity of culprit lesions (CLs) correlates with that of non-culprit lesions (NCLs) and with systemic inflammation.MethodsWe included DM patients who had two angiographically significant lesions and were undergoing percutaneous coronary intervention. We measured the temperature difference (ΔT) between the lesion and proximal vessel wall.ResultsWe included 104 (n = 208 lesions) patients: 32 (n = 64 lesions) had DM and 72 (n = 144 lesions) were non-DM (control group). ΔT was increased in DM in both CLs and NCLs (CLs: DM = 0.12 ± 0.06 °C; no DM = 0.06 ± 0.04 °C; P < 0.01 versus NCLs: DM = 0.13 ± 0.08 °C versus no DM = 0.06 ± 0.05 °C; P < 0.01). Patients with DM had similar ΔT in CLs and NCLs (P = 0.49). A linear correlation was detected between heat production in all lesions and CRP (R = 0.45; P < 0.01), which was attributed to the correlation of ΔT in lesions of patients with DM and CRP (R = 0.32; P < 0.01). In lesions of patients with low CRP, a greater rate of discrepancy was found, as 100% of lesions in patients with DM versus 66.1% of lesions of patients without DM had a high ΔT in one or both lesions (P < 0.01).ConclusionIn patients with DM, local inflammatory activation is diffuse and correlates with systemic inflammation. However, low systemic inflammatory activation does not always predict an increase in local thermal heterogeneity.     

Risk factors of renal failure and severe complications in patients with emphysematous pyelonephritis-a single-center 15-year experience

IntroductionEmphysematous pyelonephritis (EPN) is a rare but severe infection of renal parenchyma. Risk factors of renal failure in patients survived from EPN are not clear.MethodsThe authors retrospectively reviewed the patients with a diagnosis of EPN at Taipei Veterans General Hospital from January 1, 1995, to December 31, 2009. The authors analyzed the demographic, characteristics and the treatment modalities of those patients. The renal function of survivors after EPN episode had been followed for 1 year.ResultsA total of 23 patients with a mean age of 62.8 ± 17.1 years were enrolled. Mean hospital duration was 31.8 ± 21.6 days. Fifteen (65.2%) patients had a history of diabetes mellitus. Mean serum HbA1c level among the diabetic patients was 11.7 ± 3.3. More than half of patients had Escherichia coli in culture. Eleven (47.8%) patients received antibiotic treatment alone. Twelve (52.2%) patients received the concurrent percutaneous drainage and antibiotics. The overall mortality rate was 8.6%. Shock, long hospital duration and the extensive classes of computed tomography image were correlated with poor outcome. A higher initial serum creatinine level (2.8 ± 1.4 versus 1.6 ± 0.8, P = 0.015) and receiving invasive therapy (67.7% versus 12.5%, P = 0.017) significantly contributed to chronic kidney disease in the follow-up. Shock is also an independent predictor of the poor outcome in those patients (P = 0.026).ConclusionsIn the current era, antibiotics alone provide a high success rate for the treatment of EPN. Invasive therapy is a predictor of development of chronic kidney disease. Initial resuscitation and antibiotic therapy are still the cornerstone and have the benefit of the preservation of renal function.     

Effects of short-term low- and high-carbohydrate diets on postprandial metabolism in non-diabetic and diabetic subjects

Background and aimLow-fat high-carbohydrate diets raise plasma triacylglycerol (TG) concentrations. To test whether the nature of the carbohydrate affects metabolic responses, we conducted a randomized cross-over study using a short-term, intensive dietary modification.Methods and resultsEight non-diabetic subjects and four subjects with diet-controlled type 2 diabetes participated. They followed three isoenergetic diets, each for 3 days: high-fat (50% energy from fat), high-starch and high-sugar (each 70% energy from carbohydrate). Normal foods were provided. We measured plasma TG and glucose concentrations, fasting and after a standard test meal, on day 4 following each dietary period. Fasting TG concentrations were greatest following the high-sugar diet (mean ± SEM for all subjects 1900 ± 420 μmol/l) and lowest following high-fat (1010 ± 130 μmol/l) (P = 0.001); high-starch (mean 1500 ± 310) and high-fat did not differ significantly (P = 0.06). There was a greater effect in the diabetic subjects (diet × diabetes status interaction, P = 0.008). Postprandial TG concentrations were similarly affected by prior diet (P < 0.001) with each diet different from the others (P ≤ 0.01). The elevation of fasting TG on the high-sugar versus high-fat diet was strongly related to the average fasting TG concentration (P = 0.01 across both diabetic and non-diabetic subjects). Fasting glucose concentrations were not affected by prior diet but postprandial glucose concentrations were (P = 0.018), with significantly higher values after the high-fat than the high-sugar diet (P = 0.03).ConclusionsThe short-term TG-raising effect of a very low-fat diet is dependent upon the nature of the carbohydrate, with a greater effect of a sugar-rich than a complex-carbohydrate-rich diet.     

Electrocardiographic findings in patients with primary dysmenorrhea

IntroductionPrimary dysmenorrhea (PD), which is characterized by painful menstrual cycles, is one of the common clinical problems in young adult women. The aim of this study was to investigate the risk of cardiac arrhythmias in PD patients by using the electrocardiographic (ECG) parameters.MethodsForty patients diagnosed with PD and 30 age-matched normal controls were included in this study. ECGs were performed by using 12-leads with 10 mV amplitude and 25 mm/sec velocity. P and QT waves were manually marked along the isoelectric line. P maximum, P minimum, QT maximum and QT minimum were measured on the surface 12-leads ECG, and the P wave and QT dispersions were calculated.ResultsThere was not any significant correlation of P wave dispersion and QT dispersion between the age, sex, body mass index, hemoglobin, fasting blood glucose or any other laboratory parameters. P wave dispersion was significantly longer in the PD group than the control group (61.4 ± 19 msec versus 57 ± 14 msec, P = 0.01). The P minimum duration was significantly shorter in the PD group compared with the control group (36 ± 16 msec versus 41 ± 9 msec, P = 0.03). QT dispersion was significantly higher in the PD group compared with normal controls (76 ± 23 msec versus 58 ± 16 msec, P = 0.02).ConclusionThese results show that PD can be associated with cardiac arrhythmias, especially atrial fibrillation, by increasing P wave dispersion and ventricular arrhythmia risk because of an increased QT interval.     

Placenta growth factor expression in human atherosclerotic carotid plaques is related to plaque destabilization

Background and purposePlacenta growth factor (PlGF) mediates angiogenesis and inflammation, but its role in human atherosclerosis is unknown. This study was designed to test the hypothesis that PlGF-expression in human atherosclerotic carotid plaques is related to inflammation, vascularization and clinical plaque instability.MethodsThe expression of PlGF, C-reactive protein (CRP) and CD40L was analyzed with Western blots in carotid plaques of 60 patients. Cellular infiltration (CD68, CD3) and vascularization (von-Willebrand-factor) was assessed by immunohistochemistry.ResultsSymptomatic patients showed higher levels of PlGF than asymptomatic patients (115.4 ± 8.2 versus 83.6 ± 10.5 densitometric units (DU), p < 0.05) and higher grading for inflammatory cells and microvessels (CD3: 2.3 ± 0.1 versus 0.6 ± 0.1, p < 0.001, CD68: 2.4 ± 0.1 versus 0.8 ± 0.1, p < 0.001, microvessels: 2.3 ± 0.1 versus 1.5 ± 0.1, p < 0.01). PlGF-expression showed a positive correlation to the expression of CRP (r = 0.5, p < 0.001) and CD40L (r = 0.4, p < 0.01).ConclusionsPlGF-expression within human atherosclerotic lesions is associated with plaque inflammation and microvascular density, suggesting a role for PlGF in plaque destabilization and, thus, in clinical manifestation of the disease.     

The implication of obesity on total antioxidant capacity in apparently healthy men and women: the ATTICA study

Background and aimWe evaluated the association of obesity with serum total antioxidant capacity (TAC), in a population-based sample of 3042 adults.Methods and resultsDuring 2001–2002 we randomly enrolled 1514 men (18–87 years old) and 1528 women (18–89 years old), from the Attica area in Greece into the study, and the sample was stratified by the age-sex distribution of the region (census 2001). Among several variables we also measured serum TAC and weight, height, waist and hip circumferences. Waist circumference greater than 102 cm for men and 88 cm for women was considered an indicator of central fat.Methods and resultsMean waist circumference was 98 ± 13 cm in men and 84 ± 22 cm in women (P < 0.001), while mean hip circumference was 106 ± 28 cm in men and 103 ± 13 cm in women (P < 0.001). Central fat prevailed in 53% of men and 45% of women (P < 0.001). Male participants with central fat exhibited 5% lower TAC concentrations compared to leaner individuals (214 ± 35 vs. 226 ± 33 μmol/L, P = 0.04) and female participants with central fat exhibited 7% lower TAC concentrations (256 ± 38 vs. 239 ± 27 μmol/L, P = 0.03). Similarly, obese or overweight male participants had 6% lower TAC concentrations compared to normal weight (217 ± 33 vs. 234 ± 39 μmol/L, P = 0.03) and female obese or overweight participants had 10% lower TAC concentrations (226 ± 32 vs. 250 ± 30 μmol/L, P = 0.02) compared to the others.ConclusionsOur results suggest an inverse relationship between body fat, central adiposity and antioxidant capacity, irrespective of age and various other potential confounders, namely smoking, physical activity, dietary habits, blood pressure, glucose levels, and lipid concentrations.     

Unstable shoes increase energy expenditure of obese patients

BackgroundErgonomic unstable shoes, which are widely available to the general population, could increase daily non-exercise activity thermogenesis as the result of increased muscular involvement. We compared the energy expenditure of obese patients during standing and walking with conventional flat-bottomed shoes versus unstable shoes.MethodsTwenty-nine obese patients were asked to stand quietly and to walk at their preferred walking speed while wearing unstable or conventional shoes. The main outcome measures were metabolic rate of standing and gross and net energy cost of walking, as assessed with indirect calorimetry.ResultsMetabolic rate of standing was higher while wearing unstable shoes compared with conventional shoes (1.11 ± 0.20 W/kg^?1vs 1.06 ± 0.23 W/kg^?1, P = .0098). Gross and net energy cost of walking were higher while wearing unstable shoes compared with conventional shoes (gross: 4.20 ± 0.42 J/kg^?1/m^?1vs 4.01 ± 0.39 J/kg^?1/m^?1, P = .0035; net: 3.37 ± 0.41 J/kg^?1/m^?1vs 3.21 ± 0.37 J/kg^?1/m^?1; P = .032).ConclusionIn obese patients, it is possible to increase energy expenditure of standing and walking by means of ergonomic unstable footwear. Long-term use of unstable shoes may eventually prevent a positive energy balance.     

Relationship between serum levels of osteocalcin and atherosclerotic disease in type 2 diabetes

AimsTo analyze the relationship between serum levels of osteocalcin and parameters of atherosclerosis in patients with type 2 diabetes mellitus (T2DM).MethodsThis cross-sectional study of 78 patients with T2DM evaluated intima–media thickness, and the prevalence of coronary heart disease, atherosclerotic plaques and aortic calcifications. Serum osteocalcin levels were also determined by radioimmunoassay.ResultsThe patients’ mean age was 57.8 ± 6.4 years (duration of diabetes: 13.4 years; mean HbA_1c level: 8.01%), and 37.2% had coronary heart disease, 56% had an abnormal intima–media thickness, 26.9% had carotid plaques and 32.1% had aortic calcifications. Coronary heart disease was associated with higher levels of osteocalcin in male vs female patients (1.95 ± 1.36 vs 0.93 ± 0.86 ng/mL, respectively; P = 0.006). Also, higher concentrations of osteocalcin were found in female patients with vs without abnormal intima–media thicknesses (2.17 ± 1.84 vs 1.25 ± 0.67 ng/mL, respectively; P = 0.042), carotid plaques (2.86 ± 2.10 vs 1.43 ± 1.09 ng/mL, respectively; P = 0.03) and aortic calcifications (2.85 ± 1.97 vs 1.26 ± 0.83 ng/mL, respectively; P = 0.002). Serum osteocalcin levels were associated with coronary heart disease on multivariate logistic regression (odds ratio: 2.27, 95% confidence interval: 1.21–4.25; P = 0.01).ConclusionIn T2DM patients, serum osteocalcin levels were associated with parameters of atherosclerosis, suggesting that osteocalcin is involved not only in bone metabolism, but also in atherosclerotic disease.     

Effect of everolimus on pre-existing atherosclerosis in LDL-receptor deficient mice

ObjectiveProliferation signal inhibitors/mTOR-inhibitors have been shown to reduce de novo development of hypercholesterolemic atherosclerosis in animal models. However, their effect on pre-existing atherosclerosis has not yet been studied.Methods and resultsFeeding LDL-R-KO mice a high cholesterol diet for 12 weeks resulted in formation of moderate fibroatheroma (induction phase). Sixty mice received either everolimus (1 or 5 mg/kg) or no everolimus for further 12 weeks (treatment phase). Everolimus significantly enhanced hypercholesterolemia (plasma cholesterol +45%, p < 0.001). Atherosclerosis progressed obstructively in treated and non-treated mice. Everolimus (5 mg/kg) tended to reduced progression in aortic root lesions (0.28 ± 0.02 vs. 0.33 ± 0.03 mm², p = ns) and brachiocephalic lesions (0.044 ± 0.006 vs. 0.066 ± 0.012 mm², p = ns) but without significance. Everolimus (5 mg/kg) resulted in an arrest of CD68 positive plaque area (p = 0.03) and nearly halved CD68 fraction (p = 0.05) in aortic root lesions but not in brachiocephalic lesions. Taken together, despite a trend to reduced progression and inflammatory cell content there was less conclusive net effect of everolimus treatment than expected.ConclusionA higher potential of everolimus in the treatment of atherosclerosis might be obscured by its concomitant hypercholesterolemia. Considering stronger effects in previous studies we suggest that everolimus might exert more potent anti-atherogenic properties in earlier stages of atherogenesis than in advanced atherosclerosis.     

Plaque rupture and morphological characteristics of the culprit lesion in acute coronary syndromes without significant angiographic lesion: analysis by intravascular ultrasound

PurposeTo evaluate by intravascular ultrasound (IVUS) the characteristics of the culprit lesion with plaque rupture without significant angiographic stenosis after acute coronary syndromes (ACS).Patients and methodsAfter ACS, IVUS was performed in 68 patients (46.8 years ± 11.9) without significant angiographic stenosis (31 ± 15%). Plaque rupture was defined as a cavity within the plaque, communicating with the arterial lumen and having an overlying residual fibrous cap fragment. Qualitative analysis defined the type of plaque, and quantitative analysis evaluated plaque plus media area, plaque volume, plaque burden, and arterial remodeling index. Patients were divided into two groups: Group I with plaque rupture (25 patients) and Group II without plaque rupture (43 patients).ResultsAll patients with rupture showed soft or mixed plaque but no calcified plaque. In Group I, plaque rupture was associated with a larger plaque burden (49.8 ± 12.3% vs. 39.8 ± 12.1%, P < .0005), a more significant plaque plus media area (7.44 ± 2.9 vs. 5.24 ± 2.4 mm², P < .001), a greater plaque volume (151.9 ± 103.4 vs. 99.2 ± 81.6 mm³, P < .007), and a higher ratio of plaque volume over length (8.0 ± 3.8 vs. 5.6 ± 3.7 mm³/mm, P < .003). In Group I, positive remodeling was more frequent than intermediate remodeling (P < .03) or negative remodeling (P < .005). In Group II, there was no significant difference between the three types of remodeling.ConclusionThe plaque ruptures responsible for ACS frequently appear on voluminous plaques with a large plaque burden and positive arterial remodeling.     

Long-term use of oral nicorandil stabilizes coronary plaque in patients with stable angina pectoris

ObjectiveThe Impact of Nicorandil in Angina (IONA) trial demonstrated that the use of nicorandil, an anti-anginal drug, reduced future cardiovascular events in patients with stable angina. We hypothesized that nicorandil has beneficial effects on coronary arterial plaque characteristics and atherosclerogenesis.Methods and ResultsPreintervention intravascular ultrasound-virtual histology was performed prospectively in 65 consecutive patients with stable angina pectoris. There were no differences in coronary risk factors between the nicorandil (n = 16) and non-nicorandil (n = 49) groups. However, the nicorandil group demonstrated a larger %fibrous tissue (68 ± 10 vs. 62 ± 11%, P = 0.049) and a smaller %necrotic core tissue (11 ± 7 vs. 16 ± 10%, P = 0.049) compared with the non-nicorandil group. Multiple regression analysis showed that %necrotic core tissue (P = 0.045) was negatively and %fibrous tissue (P = 0.026) was positively associated with the use of nicorandil independent of statin use. We also analyzed the effect of nicorandil on atherosclerotic lesion formation in a mouse model of atherosclerosis. Lipid profiles were unaffected, but the area of atherosclerotic lesion and plaque necrosis were significantly reduced following 8-week nicorandil treatment in ApoE-deficient mice fed an atherogenic diet. Nicorandil significantly reduced the expression levels of endoplasmic reticulum stress markers, C/EBP homologous protein (CHOP) and glucose regulated protein/BiP (GRP78) in atherosclerotic lesions. Nicorandil significantly attenuated tunicamycin-induced CHOP upregulation in cultured THP-1 macrophages.ConclusionsNicorandil exerts its anti-atherogenic effect by mechanisms different from those of statins. Long-term nicorandil treatment is a potentially suitable second-line prevention therapy for patients with coronary artery disease.     

Serum amyloid A may potentiate prothrombotic and proinflammatory events in acute coronary syndromes

AimsElevated serum amyloid A (SAA) levels, like C-reactive protein (CRP), predict coronary events. Both induce monocyte tissue factor (TF), and peripheral blood mononuclear cells (PBMC) from patients with coronary artery disease (CAD) express higher TF in response to CRP. This study examined SAA induction of TF and tumour necrosis factor-α (TNF) in PBMC from patients with CAD and in monocytoid THP-1 cells.Methods and resultsPBMC from 26 males with CAD (15 stable angina, SA, and 11 acute coronary syndromes, ACS) and 14 male controls were stimulated with SAA. SAA promoted up to six-fold increase in TF activity (recalcification assay) on PBMC from patients, associated with elevated TF mRNA and protein. PBMC responded optimally when monocytes were adherent. Unlike CRP, SAA induced TF and TNF in THP-1 cells. SAA-induced TNF was dose-dependently inhibited by HDL. PBMC from patients with ACS expressed more basal TF (257.4 ± 46.8 mU/10⁶ PBMC vs. 131.0 ± 12.5 mU/10⁶ PBMC, P = 0.003), and greater SAA-induced TF than cells from controls, whereas no difference was found between SA and controls (ACS 2246 ± 493, SA 1364 ± 206, controls 1091 ± 113 mU/10⁶ PBMC, with SAA 250 ng/mL, P = 0.002 ACS vs. controls across the dose range). Importantly, SAA-induced TNF levels (ELISA) were much higher in patients with ACS than SA or controls (ACS 211 ± 41, SA 108 ± 16, controls 73 ± 11 pg/mL, with SAA 250 ng/mL, P = 0.001 ACS vs. controls or P = 0.013 ACS vs. SA across the dose range). SAA-induced TF and TNF correlated positively with serum SAA levels in CAD, but not controls.ConclusionsSAA is a prothrombotic and proinflammatory mediator in ACS which may contribute to atherogenesis and its complications.     

Propylthiouracil, independent of its antithyroid effect, produces endothelium-dependent vasodilatation through induction of nitric oxide bioactivity

ObjectivePropylthiouracil (PTU), independent of its antithyroid effect, is recently found to have a potent antiatherosclerotic effect. The aim of this study is to investigate whether PTU has a beneficial effect on endothelial function.Methods and resultsNinety patients with a history of hyperthyroidism receiving either PTU (n = 45) or methimazole (MMI) (n = 45) during the euthyroid status were enrolled in this study. Brachial artery endothelium-dependent (flow-mediated dilatation [FMD]) and endothelium-independent (nitroglycerin-mediated dilatation) responses were assessed by high-resolution ultrasound image. Data for these two groups were compared with those of 41 healthy control subjects. The FMD values were significantly increased in patients maintained on PTU versus those in the MMI and control groups (9.3 ± 4.4%, 3.4 ± 2.5%, and 3.6 ± 3.4%, respectively; P < 0.01). Nitroglycerin-mediated dilatation had no significant difference between the PTU, MMI, and control groups (17.4 ± 7.5%, 15.9 ± 6.1%, and 17.5 ± 6.8%, respectively; P = 0.455). On multivariate analysis, no significant relationship was found between the FMD and thyroid hormone index levels. To further elucidate whether PTU has a direct effect on endothelial function, the effect of PTU on isolated segments of Sprague–Dawley rat aorta was studied. Vasodilatation induced by PTU was endothelium-dependent and could be blocked by pretreatment with nitric oxide (NO) inhibitors. PTU also increased NO formation in aortic segments.ConclusionsThis study demonstrated that PTU produced endothelium-dependent vasodilatation through thyroid-independent and NO-mediated mechanisms that may contribute to its beneficial effect on atherosclerosis.     

The FTO gene modifies weight, fat mass and insulin sensitivity in women with polycystic ovary syndrome, where its role may be larger than in other phenotypes

AimGenome-wide association studies have shown that variation in the FTO gene predisposes to obesity and related traits that are common features of polycystic ovary syndrome (PCOS). The aim of the present study was to assess the effect of FTO variation on obesity, insulin sensitivity, and metabolic and hormonal profiles in PCOS.MethodsWe examined 136 PCOS women (mean body mass index [BMI]: 28.28 ± 6.95 kg/m², mean age: 25.36 ± 5.48 years). Anthropometric measurement, euglycaemic–hyperinsulinaemic clamp and oral glucose tolerance tests and sex hormone assessments were performed. The study group was genotyped for the FTO rs9939609 polymorphism.ResultsBMI (29.0 ± 6.9 kg/m² vs 26.1 ± 6.8 kg/m²; P = 0.023), body weight (80.1 ± 20.7 kg vs 72.6 ± 20.2 kg; P = 0.048), fat mass (29.7 ± 1 6.6 kg vs 24.6 ± 17.7 kg; P = 0.045) and waist circumference (89.8 ± 16.7 cm vs 83.2 ± 17.1 cm; P = 0.028) were higher in carriers of at least one copy of the A allele. Differences in these parameters were more significant when comparing AA and TT homozygotes. Women with the AA genotype also had decreased insulin sensitivity (P = 0.025) and follicle-stimulating hormone (P = 0.036). In logistic-regression analyses, the association of the FTO gene polymorphism with insulin sensitivity was no longer significant when BMI was included in the model.ConclusionVariation in the FTO gene modifies weight, adiposity and other measures of obesity and insulin sensitivity in PCOS. The examined FTO gene variant appears to have a greater impact on obesity and related traits in PCOS than in other phenotypes. The effect on insulin sensitivity appears to be secondary to its influence on obesity and body fat.     

Effect of treatment with human apolipoprotein A-I on atherosclerosis in uremic apolipoprotein-E deficient mice

ObjectiveUremia markedly increases the risk of atherosclerosis. Thus, effective anti-atherogenic treatments are needed for uremic patients. This study examined effects of non-lipidated recombinant human apoA-I (h-apoA-I) and a recombinant trimeric apoA-I molecule (TripA-I) on lipid metabolism and atherosclerosis in uremic apoE?/? mice.Methods and resultsUpon intraperitoneal injection, h-apoA-I and TripA-I rapidly associated with plasma HDL and reduced mouse apoA-I plasma levels without affecting total or HDL cholesterol concentrations. The plasma half-life was ～36 h for TripA-I and ～16 h for h-apoA-I. Injection of h-apoA-I (100 mg/kg) or TripA-I (100 mg/kg) twice weekly for 7 weeks did not affect the cross-sectional area of atherosclerotic lesions in the aortic root, or the en face lesion area and cholesterol content in the thoracic aorta in uremic apoE?/? mice. Also, the treatments did not affect expression of selected inflammatory genes in the thoracic aorta or plasma concentrations of soluble ICAM-1 and VCAM-1. However, h-apoA-I-treated mice had larger smooth muscle cell-staining areas in aortic root plaques than PBS-treated mice (4.8 ± 0.8% vs. 2.5 ± 0.6%, P < 0.05).ConclusionsThe data suggest that long-term treatment with non-lipidated h-apoA-I or TripA-I might affect plaque composition but does not reduce atherosclerotic lesion size in uremic apoE?/? mice.     

Measurement of Rho-associated kinase (ROCK) activity in humans: validity of leukocyte p-MBS/t-MBS in comparison with vascular response to fasudil

BackgroundRho-associated kinases (ROCKs) have been shown to be involved in the pathogenesis of atherosclerosis. It is clinically important to estimate the degree of ROCK activity in humans. The purpose of this study was to confirm the validity of a leukocyte ROCK parameter as an index of ROCK activity in comparison with vascular response to a ROCK inhibitor.Methods and resultsWe evaluated the ratio of phospho myosin-binding subunit (p-MBS) on myosin light-chain phosphatase to total MBS in peripheral leukocytes by Western blot analysis and forearm blood flow (FBF) response to the ROCK inhibitor fasudil using strain-gauge plethysmography in 36 healthy subjects and 39 patients with cardiovascular diseases. Fasudil (3, 10, 30 μg/min) was infused intra-arterially for 5 min at each dose. Leukocyte p-MBS/total-MBS was higher in cardiovascular diseases than in healthy subjects (0.97 ± 0.37 vs. 0.51 ± 0.14; P = 0.002). Fasudil increased FBF from 4.9 ± 1.2 to 14.5 ± 5.7 mL/min/100 mL tissue (P < 0.0001) in patients with cardiovascular diseases, while fasudil did not alter FBF in healthy subjects. There was a significant relationship between leukocyte p-MBS/total-MBS and maximal FBF response to fasudil in all subjects (r = 0.72, P < 0.0001). There was also a significant correlation between p-MBS/total-MBS and maximal FBF response to fasudil in patients with cardiovascular diseases (r = 0.59, P < 0.0001). In healthy subjects, there was no significant correlation between the two parameters.ConclusionsThese findings suggest that assessment of leukocyte ROCK activity is minimally invasive and does not require pharmacologic intervention using ROCK inhibitors. Leukocyte p-MBS/total-MBS may be useful for evaluating ROCK activity in a clinical setting.     

Digitoxin Prolongs Survival of Female Rats With Heart Failure Due to Large Myocardial Infarction

BackgroundWe analyzed whether digitoxin affects the survival of rats with congestive heart failure.Methods and ResultsThe influence of digitoxin (0.1 mg·100 g·day, orally) on the survival of infarcted female rats (n = 170) randomized as Control Infarcted (CI, n = 85) or Digitoxin (D, n = 85) was evaluated for 280 days. Mean survival was 235 ± 7 days for CI and 255 ± 5 days for D (log-rank test: P = .0602). Digitoxin did not affect survival in rats with congestive heart failure from myocardial infarction <40% of the left ventricle, but did prolong survival in rats with infarction ≥40%. The log-rank test defined higher mortality (P = .0161) in CI >40% (56%) than in D >40% (34%), with a hazard ratio of 2.03. Pulmonary water content and papillary muscle mechanics were analyzed in CI (n = 7) and D (n = 14) survivors. Significant differences were observed regarding pulmonary water content (CI: 82 ± 0.3; D: 80 ± 0.3%; P = .0014), developed tension (CI: 2.7 ± 0.3; D: 3.8 ± 0.3 g/mm²; P = .0286) and +dT/dt (CI: 24 ± 3; D: 39 ± 4 mg mm²·s; P = .0109).ConclusionIn conclusion, long-term digitoxin administration reduced cardiac impairment after myocardium infarction, attenuated myocardial dysfunction, reduced pulmonary congestion, and provided the first evidence regarding the efficiency of digitoxin in prolonging survival in experimental cardiac failure.     

Circulating angiogenic cell populations, vascular function, and arterial stiffness

ObjectiveSeveral bone marrow-derived cell populations have been identified that may possess angiogenic activity and contribute to vascular homeostasis in experimental studies. We examined the extent to which lower quantities of these circulating angiogenic cell phenotypes may be related to impaired vascular function and greater arterial stiffness.MethodsWe studied 1948 Framingham Heart Study participants (mean age, 66 ± 9 years; 54% women) who were phenotyped for circulating angiogenic cells: CD34+, CD34+/KDR+, and early outgrowth colony forming units (CFU). Participants underwent non-invasive assessments of vascular function including peripheral arterial tone (PAT), arterial tonometry, and brachial reactivity testing.ResultsIn unadjusted analyses, higher CD34+ and CD34+/KDR+ concentrations were modestly associated with lower PAT ratio (β = ?0.052 ± 0.011, P < 0.001 and β = ?0.030 ± 0.011, P = 0.008, respectively) and with higher carotid-brachial pulse wave velocity (β = 0.144 ± 0.043, P = 0.001 and β = 0.112 ± 0.043, P = 0.009), but not with flow-mediated dilation; higher CD34+ was also associated with lower carotid-femoral pulse wave velocity (β = ?0.229 ± 0.094, P = 0.015). However, only the association of lower CD34+ concentration with higher PAT ratio persisted in multivariable analyses that adjusted for standard cardiovascular risk factors. In all analyses, CFU was not associated with measures of vascular function or arterial stiffness.ConclusionsIn our large, community-based sample of men and women, circulating angiogenic cell phenotypes largely were not associated with measures of vascular function or arterial stiffness in analyses adjusting for traditional risk factors.     

Effect of an oral astaxanthin prodrug (CDX-085) on lipoprotein levels and progression of atherosclerosis in LDLR(-/-) and ApoE(-/-) mice

Oxidative stress and inflammation are key promoters of atherosclerosis and myocardial damage. When orally administered, the novel astaxanthin prodrug CDX-085 delivers high levels of the xanthophyll antioxidant astaxanthin that protects LDL from oxidation and reduces primary thrombosis. In this study, we analyzed whether delivery of astaxanthin from administration of the CDX-085 prodrug reduces plasma lipoprotein levels and the progression of atherosclerosis in low-density lipoprotein receptor negative (LDLR^?/?) and apolipoprotein E deficient (ApoE^?/?) mice.MethodsRelative circulating levels of astaxanthin derived from CDX-085 administration compared to administration of pure astaxanthin was initially evaluated in a canine model. In mouse Study #1, 16 wild-type and 16 LDLR^?/? mice on 0.5% cholesterol diet supplemented with either 0.0%, 0.08%, 0.2% and 0.4% CDX-085 were used to assess plasma levels and lipoprotein biodistribution measured by FPLC after 4 weeks treatment. In Study #2, 36 male LDLR^?/? mice were randomized to a 0.5% cholesterol chow diet (CHOW group, n = 12) or 0.5% cholesterol chow fortified with 0.08% CDX-085 (n = 12) or 0.5% cholesterol chow with 0.4% CDX-085 (n = 12) for 12 weeks. In Study #3, 34 male ApoE^?/? mice were randomized in the same fashion as the Study #2 and fed similar diets for 9 weeks.ResultsCDX-085 administration was shown to result in significantly higher levels of circulating astaxanthin (p < 0.001 ANOVA) over a 72 h period compared to pure, non-esterified astaxanthin in a single-dose pharmacokinetic study in beagles. In Study #1, plasma astaxanthin levels were 5–9-fold higher in LDLR^?/? mice compared to wild-type mice. Astaxanthin was highly distributed among all lipoprotein fractions, generally reflecting cholesterol content of lipoproteins. In Study #2, administration of CDX-085 resulted in significantly lower total cholesterol levels (528 ± 68 mg/dL vs. 550 ± 67 mg/dL vs. 602 ± 80 mg/dL, p = 0.047) and aortic arch atherosclerosis (9.0 ± 4.2% vs. 9.8 ± 3.5% vs. 13.2 ± 3.6%, p = 0.023) in the 0.4% CDX-085 group compared to the 0.08% CDX-085 and CHOW groups, respectively. In ApoE^?/? mice, a 72% reduction in triglycerides in the 0.4% CDX-085 group and 50% reduction in the 0.08% CDX-085 groups was noted compared to CHOW group (final levels 17 ± 11 mg/dL vs. 30 ± 15 mg/dL vs. 60 ± 32 mg/dL, respectively, p = 0.001).ConclusionOral administration of the novel astaxanthin prodrug CDX-085 shows that it distributes among lipoproteins. CDX-085 lowers total cholesterol and aortic arch atherosclerosis in LDLR^?/? mice and triglyceride levels in ApoE^?/? mice and shows promise for further evaluation in human studies.