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Objectives: To evaluate the association between inflammation, oxidative stress, and circulating progenitor cell (CPC) number and redox equilibrium, vascular lesions and accelerated atherosclerosis in rheumatoid arthritis (RA). Method: Circulating CD34+ cells were isolated from 33 RA patients and 33 controls. Reactive oxygen species (ROS) levels and mRNA expression of manganese superoxide dismutase (MnSOD), catalase (CAT), glutathione peroxidase type 1 (GPx-1) antioxidant enzymes, and the gp91phox-containing nicotinamide adenine dinucleotide phosphate (NADPH) oxidase NOX2 were measured in CD34+ cells. C-reactive protein (CRP), fibrinogen, erythrocyte sedimentation rate (ESR), carotid intima-media thickness (cIMT), and arterial stiffness (AS) were also evaluated. We investigated the relationships between inflammatory markers, vascular parameters, cell number, and antioxidant enzymes. Results: CD34+ cell number was lower in RA patients than in controls. In CD34+ cells from RA patients, ROS levels, MnSOD mRNA, and NOX2 mRNA were higher, while mRNA expression of GPx-1 and CAT was significantly lower. The AS, pulse wave velocity (PWV), and augmentation index (AIx) were higher, as was cIMT. CD34+ cell number was inversely correlated with CRP, ROS, PWV, and AIx, and with the CAT/MnSOD and GPx-1/MnSOD ratios. CRP was correlated with MnSOD mRNA, PWV, and AIx but not with CAT and GPx-1 mRNA. Conclusions: Our data show a link between inflammation, oxidative stress, and the impairment of the antioxidant system of CPCs and their number, and with arterial stiffness in RA subjects. This could suggest a perspective on the accelerated development of vascular damage and atherosclerosis in RA. [ABSTRACT FROM AUTHOR]

BACKGROUND AND AIMS: Aging is associated with an increased risk of developing atherosclerosis. Subjects over 80 years of age without cardiovascular disease provide a model to investigate the protective factors increasing their resistance to atherosclerotic disease. Platelet-activating factor acetylhydrolase (PAF-AH) is an enzyme associated with low density lipoprotein (LDL) and high density lipoprotein (HDL) inactivating plateletactivating factor (PAF) and preventing LDL oxidation by hydrolysis of oxidized phospholipids. The aim of the present study was to evaluate the contribution of the PAFAH gene Arg92His, Ile198Thr and Ala379Val polymorphisms to resistance toward developing cardiovascular events in healthy Sicilian octogenarians. METHODS: Distribution of PAF-AH genotypes and activity, and biochemical parameters, were compared between 100 octogenarians and 200 healthy adults. RESULTS: The individuals in the elderly group displayed significantly higher levels of HDL-C (p<0.001) and plasma (p<0.001) and HDL (p<0.001) PAF-AH activity. Analysis of PAFAH genotype distributions showed no significant differences between octogenarians and controls. No differences among PAF-AH genotypes with respect to plasma and HDL PAF-AH activity were found in either group. CONCLUSIONS: Our results provide no evidence of a significant association between the PAF-AH gene Arg92His, Ile198Thr and Ala379Val polymorphisms and successful aging in Sicilians. They also emphasize that, in these subjects, aging is characterized by increased levels of PAF-AH activity and HDL-C. [ABSTRACT FROM AUTHOR]

This study was designed to determine whether the levels of soluble intercellular adhesion molecule-1 (sICAM-1) and tumor necrosis factor-alpha (TNF-alpha) were elevated in subjects with uncomplicated hypertension who presented with no other risk factors or evidence of athero-sclerosis. The effects of administration of an angiotensin type-1 antagonist (losartan) on the serum concentrations of these molecules were also examined. Twenty hypertensive (HT) subjects (12 men and 8 women, mean age 49.1 +/-7.2 years) without other risk factors or cardiovascular disease received placebo for 4 weeks. The patients were then treated with losartan (50 mg/day) for 24 weeks. After 4, 12, and 24 weeks of losartan treatment, sICAM-1 and TNF-alpha levels were measured. The same parameters were measured in 20 normotensive control subjects (C), matched for sex and age. HT had sICAM-1 and TNF-alpha basal values higher than C (respectively 351.7 +/-97.4 vs 201.6 +/-32.3 ng/mL, p<0.001 and 31.8 +/-2.4 vs 15.3 +/-2.2 pg/mL, p<0.001). There was a positive correlation between sICAM-1 and TNF-alpha levels, but no correlation in HT between the average diastolic and systolic blood pressure (clinic and ambulatory monitoring) and the sICAM-1 or TNF-alpha levels was observed. Losartan treatment caused a significant decrease of sICAM-1 levels at the end of the first month of treatment (300.2 +/-64.4 ng/mL, p<0.05), but the values reverted to the basal levels at the following time points. No variation of TNF-alpha levels during losartan treatment was observed. These results show that patients with uncomplicated mild essential hypertension presented with high plasma ICAM-1 and TNF-alpha concentrations. Although all the patients were responsive to the antihypertensive treatment with losartan, their plasma concentrations of TNF-alpha were not modified, and sICAM-1 concentrations decreased only for a short period of time. This suggests that in uncomplicated hypertension other factors besides the blood pressure modulate the endothelial inflammation. [ABSTRACT FROM AUTHOR]

Objective. To identify, evaluate, and synthesize evidence on the predictive power of circulating endothelial progenitor cells (EPCs) in cardiovascular disease, through a systematic review of quantitative studies. Data Sources. MEDLINE was searched using keywords related to "endothelial progenitor cells" and "endothelium" and, for the different categories, respectively, "smoking"; "blood pressure"; "diabetes mellitus" or "insulin resistance"; "dyslipidemia"; "aging" or "elderly"; "angina pectoris" or "myocardial infarction"; "stroke" or "cerebrovascular disease"; "homocysteine"; "C-reactive protein"; "vitamin D". Study Selection. Database hits were evaluated against explicit inclusion criteria. From 927 database hits, 43 quantitative studies were included. Data Syntheses. EPC count has been suggested for cardiovascular risk estimation in the clinical practice, since it is currently accepted that EPCs can work as proangiogenic support cells, maintaining their importance as regenerative/reparative potential, and also as prognostic markers. Conclusions. EPCs showed an important role in identifying cardiovascular risk conditions, and to suggest their evaluation as predictor of outcomes appears to be reasonable in different defined clinical settings. Due to their capability of proliferation, circulation, and the development of functional progeny, great interest has been directed to therapeutic use of progenitor cells in atherosclerotic diseases. This trial is registered with registration number: Prospero CRD42015023717.

Hypothesis/introduction: The aim of this study was to assess the antihypertensive efficacy and safety of aliskiren versus ramipril or losartan in hypertensive patients with type 2 diabetes mellitus, microalbuminuria and uncontrolled hypertension, despite the use of optimal conventional antihypertensive therapy., Materials and Methods: In this open-label active comparator study, 126 patients were randomly assigned to receive 24 weeks of additional therapy with aliskiren (Group A) or either losartan or ramipril (Group B), according to whether a patient was already treated with an angiotensin-converting enzyme inhibitor or an angiotensin receptor blocker, respectively., Results: After 24 weeks, both treatment groups experienced a significant reduction of systolic blood pressure (-11.37% and -8.47%, respectively; both p <0.001 vs. baseline) and diastolic blood pressure levels (-10.67% and -9.28%, respectively; both p <0.001 vs. baseline), with a greater reduction of mean systolic values in Group A compared with Group B (p <0.001). Furthermore, after six months microalbuminuria was significantly decreased in both treatment groups (-67.62% and -49.1%, respectively; both p <0.001), with a reduction rate in Group A significantly higher than in Group B (p<0.001)., Conclusions: The addition of aliskiren to optimal conventional therapy provided a higher reduction of blood pressure and urinary albumin excretion when compared with the addition of losartan or ramipril., (© The Author(s) 2014.)

Background and Aims: Suboptimal vitamin D status was recently acknowledged as an independent predictor of cardiovascular diseases and all-cause mortality in several clinical settings, and its serum levels are commonly reduced in Rheumatoid Arthritis (RA). Patients affected by RA present accelerated atherosclerosis and increased cardiovascular morbidity and mortality with respect to the general population. In RA, it has been reported an impairment of the number and the activity of circulating proangiogenic haematopoietic cells (PHCs), including CD34+, that may play a role in endothelial homeostasis. The purpose of the study is to investigate the association between vitamin D levels and PHCs, inflammatory markers, and arterial stiffening in patients with RA., Methods and Results: CD34+ cells were isolated from 27 RA patients and 41 controls. Vitamin D levels, C-reactive protein (CRP), fibrinogen, pulse wave velocity (PWV), and carotid intima-media thickness (cIMT) were also evaluated. CD34+ count and vitamin D levels were lower in RA patients as compared to controls, while fibrinogen, CRP, PWV and cIMT were higher in RA patients. CD34+ cell number appeared to be associated with vitamin D levels, and negatively correlated to fibrinogen and early atherosclerosis markers (PWV and cIMT); vitamin D levels appear also to be inversely associated to fibrinogen., Conclusions: RA patients with moderate disease activity presented with low vitamin D levels, low CD34+ cell count, increased PWV and cIMT; we found that vitamin D deficiency is associated to CD34+ cell reduction in peripheral blood, and with fibrinogen levels. This suggests that vitamin D might contribute to endothelial homeostasis in patients with RA.

Objective: Cigarette smokers present early signs of vascular damage and systemic inflammation. Biglycan (BGN), an ubiquitous component of extracellular matrix orchestrating several physiological functions, has recently been indicated as a major source of low-density lipoprotein retention in the normal arterial intima-media layer. We evaluated whether BGN-mRNA expression was enhanced in peripheral monocytes of smokers with no additional cardiovascular risk factors (CVRFs), and if it was associated with altered carotid arterial stiffness (AS) or intima media thickness (cIMT). We also evaluated plasma markers of systemic and vascular inflammation, and correlation with BGN-mRNA., Methods: Two-hundred-fifty-one young smokers were enrolled, with no additional CVRFs, and 60 controls. Plasma lipids, fibrinogen, C-reactive protein (CRP), interleukin-6 (IL-6), AS and cIMT were assessed. A smoke exposure index (SEIx) was calculated., Results: Fibrinogen, CRP, AS indices, cIMT, and BGN-mRNA were higher in smokers compared to controls; HDL-C levels were lower, no difference was detected in IL-6 levels. After stratification of smokers in quartiles based on SEIx values, smokers in the highest quartiles presented highest fibrinogen, CRP, AS, cIMT, BGN, and also IL-6 values, and lowest HDL-C., Conclusion: BGN-mRNA was enhanced in young smokers, compared to controls, and appears associated to a proatherogenic profile, characterized by increased fibrinogen, CRP, and IL-6, lower HDL-C, altered AS and cIMT values, particularly in those with higher SEIx: the more cigarettes smoked over years, the more marked the alterations. Although we cannot state whether BGN have a direct causal role in inducing, maintaining and developing vascular damage, including intima-media wall thickening and arterial stiffening, our data could suggest that it may represent a link between proatherogenic status induced by cigarette smoking, and the development and progression of vascular damage., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Objectives: Circulating proangiogenic haematopoietic cells (PHCs), including CD34+ cells, play an important role in endothelial homeostasis. Among PHCs, CD34+ cells are the largest cell population, thus, much of the regenerative/reparative potential of PHCs may be attributed to CD34+ cells. Our aim was to determine the association between inflammation and CD34+ cell number, intracellular levels of reactive oxygen species (ROS) and expression of Toll-like receptor 3 (TLR3) and interleukin 1β (IL-1β), arterial stiffness (AS) indices, and carotid intima-media thickness (cIMT) in patients affected by rheumatoid arthritis (RA)., Methods: CD34+ cells were isolated from 24 RA patients and 26 matched controls. ROS levels, TLR3 and IL-1β expression were measured. C-reactive protein (CRP), fibrinogen, AS, and cIMT were also evaluated., Results: CD34+ count was lower in RA patients as compared to controls. In CD34+ cells from RA patients, ROS, TLR3 and IL-1β expressions were increased compared to controls. In RA patients, we found higher CRP and fibrinogen levels, and higher values of Pulse Wave Velocity (PWV) and Augmentation Index (AIx), both AS indices, and of cIMT. CD34+ cell numbers were inversely correlated with CRP, TLR3, IL-1β, ROS, and AS indices. TLR3 levels were related to CRP, IL-1β, fibrinogen and ROS. IL-1β levels were correlated with expression of CRP, ROS, and PWV., Conclusions: Inflammatory status in RA is associated with an increased expression of TLR3 and of IL-1β in CD34+ cells, which appear to affect cell number. These new findings suggest a perspective on accelerated atherosclerosis and vascular damage in RA.

We investigated whether different degrees of hypertension-related cardiovascular involvement are associated with changes in circulating proangiogenic hematopoietic cell (PHC) numbers and/or phenotypes and/or in the PHC redox system in hypertensive individuals with isolated arterial stiffening (AS) hypertensives or with both carotid intima-media thickening and left ventricular hypertrophy (LVH) hypertensives. We also evaluated microRNA (miRs) 221 and 222 (miRs221/222) expression in CD34+ cells, the relationship between these miRs and cell number and reactive oxygen species (ROS) levels, and the expression of manganese superoxide dismutase (MnSOD), catalase (CAT) glutathione peroxidase type-1 (GPx-1) and gp91phox-containing nicotinamide-adenine-dinucleotide-phosphate-oxidase (NOX2). Proangiogenic hematopoietic cells (PHCs) from hypertensive patients and controls were isolated by flow cytometry. PHCs were higher in hypertensives than in controls but were lower in LVH than in AS hypertensives. In CD34+ cells from AS hypertensives, NOX2, MnSOD, CAT and GPx-1 were overexpressed; ROS, miRs and NOX2 were also increased and were associated with cell number. In LVH, we found an imbalance in the cell redox system; MnSOD showed the highest values, whereas CAT and GPx-1 were lower than in AS hypertensives. Intracellular ROS, miRs and NOX2 were higher and inversely associated with cell number. In AS hypertensives, the redox balance may sustain the increase in PHCs; by contrast, in hypertensives with more advanced lesions, redox imbalance may result in increased oxidative stress and cell reduction.

Cardiovascular (CV) diseases and related complications are the main causes of morbidity and mortality in the elderly. CV progenitor cells, including CD34+ cells, play a role in delaying the progression of atherosclerosis. In the present study we observed 100 octogenarians for seven years, in order to address the question of whether CD34+ cell number is a predictor of longevity in selected survivors. We also checked for associations of cell expression of manganese superoxide dismutase (Mn-SOD), catalase (CAT), and glutathione peroxidase type-1 (GPx-1) antioxidative enzymes, with number of CD34+ progenitor cells and mortality. We found that in very old subjects the number of CD34+ cells at baseline were higher in subjects who reached older age at death or were still living at the end of observation period, with respect to subjects who died from all causes, including CV deaths. On the other hand, HDL-C plasma levels and, with the exception of diabetes, the classic CV risk factors (hypertension, smoking, hypercholesterolemia) showed a loss of their predictive power. A significant association between the redox system of CD34+ cells and mortality was also observed. These data suggest that, even in the elderly, CD34+ cells maintain their role in predicting mortality. CD34+ cells could thus be considered as a biomarker of longevity., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Circulating progenitor cells (CPCs), including endothelial progenitor cells (EPCs), have a key role in endothelium repair. Cellular NADPH oxidase (Nox) enzymes, including Nox-containing gp91phox, represent a source of reactive oxygen species (ROS); ROS trigger protective signals but may also have detrimental effects. Cellular defenses against ROS include the enzymes manganese superoxide dismutase (MnSOD), catalase (CAT) and glutathione peroxidase type-1 (GPx-1). We investigated the relationships of CPCs with cellular gp91phox, MnSOD, CAT, GPx-1 and ROS levels and with carotid intima-media thickness (cIMT) and stiffness in hypertensives without additional risk factors for cardiovascular disease. CPCs from 53 newly diagnosed, untreated hypertensives and from 29 controls were isolated and identified by flow cytometry. gp91phox, MnSOD, CAT, and GPx-1 mRNA and protein expression and ROS generation were evaluated in enriched samples of CD34(+) cells; cIMT and stiffness were assessed. Hypertensives showed higher arterial stiffness (P < 0.001) but no difference in cIMT with respect to controls. ROS generation was slightly increased (P=0.04), whereas gp91phox, MnSOD, CAT and GPx-1 were significantly higher (P < 0.001) with respect to controls, as was CPC number (P < 0.001), but EPCs were no different. CPC and EPC numbers correlated with gp91phox, ROS and fibrinogen (P < 0.001); moreover, gp91phox, MnSOD, CAT and GPx-1 were correlated with CPC number. In early phases of arterial hypertension, before the development of wall thickening and even in the presence of arterial mechanical impairment, CPC number may be increased to maintain an adequate number of EPCs in peripheral blood.

Background: Cigarette smoking is involved in vascular inflammation and impairment of circulating progenitor cells (CPCs), including endothelial progenitor cells (EPCs). The study aim was to evaluate the redox balance of these cells in relation to smoking exposure., Methods: Circulating cells from 36 healthy smokers and 26 controls were isolated and identified by flow cytometry. ROS generation, mRNA and protein cell expression, and enzymatic activity of MnSOD, catalase, and GPx-1 were evaluated., Results: Smokers showed higher levels of CRP and fibrinogen and lower levels of HDL-C. ROS and MnSOD were higher (p<0.001), while catalase and GPx-1 were lower (p<0.001) as was EPC number (p<0.001) in smokers. CPC and EPC correlated with HDL-C, CRP, ROS and enzyme expression and activity., Conclusions: Our data suggest that smoking exposure involves antioxidant enzymes in CPCs and EPCs and that the inflammatory response in smokers plays an important role in impairing cells and their antioxidant functions., (Copyright © 2010 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.)

1. Recently, we demonstrated that biglycan (BGN) is increased in circulating monocyte cells from hypertensive patients and that angiotensin (Ang) II is able to increase BGN expression. The present study was designed to investigate the effects of treatment with the angiotensin AT(1) receptor antagonist losartan on monocyte BGN mRNA and protein expression in essential hypertension. 2. One hundred and twenty-six newly diagnosed hypertensive patients without additional risk factors for atherosclerosis and cardiovascular disease were treated with 100 mg losartan once daily for 6 months. Biglycan mRNA and protein expression was determined in monocytes isolated from peripheral blood before (T(0)) and after (T(1)) therapy. Plasma levels of interleukin (IL)-6, tumour necrosis factor (TNF)-alpha and high sensitivity C-reactive protein (hs-CRP) were also determined. In addition, BGN mRNA and protein expression was determined after the ex vivo addition of 1 micromol/L AngII to monocytes isolated from 20 randomly selected hypertensive patients. 3. Biglycan mRNA and protein expression, blood pressure and plasma levels of fibrinogen, IL-6, TNF-alpha and CRP were significantly lower at T(1) than at T(0). Variations in BGN expression were associated with inflammatory markers, but not directly with blood pressure. In AngII-stimulated monocytes, BGN mRNA and protein expression was significantly lower at T(1) that at T(0). Moreover, mean BGN mRNA expression in AngII-stimulated monocytes isolated from losartan-treated patients was similar to baseline expression in unstimulated monocytes from untreated patients. 4. The results of the present study show that losartan can reduce BGN expression in monocytes from hypertensive patients, without any linear association with blood pressure, suggesting that the effects of AngII on BGN expression in monocytes may be modulated, in part, by an AT(1) receptor blocker.

Background: Nonalcoholic fatty liver disease (NAFLD) represents a spectrum of diseases ranging from simple steatosis to nonalcoholic steatohepatitis (NASH) and is associated with familial combined hyperlipidaemia (FCHL). Currently, the invasive liver biopsy is considered as the gold standard for evaluating liver fibrosis (LF); however, liver stiffness measurement (LSM) by transient elastography (TE) trough FibroScan device may be employed to estimate LF noninvasively. The aim of this study was to evaluate the prevalence of NAFLD in FCHL subjects and to analyse LSM with TE to better identify those individuals with a potential risk of liver disease progression., Materials and Methods: Sixty subjects with FCHL (38 men, 22 women, mean age 46.4 +/- 10.9 years) were included in the study. We studied biochemical parameters including lipid profile, glucose, transaminase and insulin; blood pressure and waist circumference (WC) were measured; BMI and HOMA-index were calculated. Ultrasonography was performed to assess liver steatosis and carotid intima-media thickness (IMT). Liver fibrosis was measured by FibroScan., Results: Patients were classified according to have no (group 0: 19%), mild (group 1: 32%) or moderate-severe (group 2: 49%) steatosis. No difference was found between group 0 and 1 concerning all study parameters. WC (P < 0.05), BMI (P < 0.05), glucose (P < 0.05), insulin (P < 0.001), HOMA-index (P < 0.001) and LSM (6.03 +/- 1.9 Kpa vs. 4.2 +/- 0.5 Kpa, P < 0.001) were significantly higher in group 2 than groups 1 and 0. Furthermore, LSM correlated with insulin (P < 0.05), glucose (P < 0.05), HOMA-index (P < 0.001), transaminase (P < 0.01) and liver steatosis (P < 0.001). Regression analysis showed that LSM (P < 0.001) and NAFLD (P < 0.01) is associated with HOMA-index; NAFLD is also associated with WC (P < 0.05)., Conclusion: Our results suggest that in FCHL subjects, HOMA-index, an insulin resistance index, is strongly associated with liver steatosis and its progression. Furthermore, in these subjects, we propose the transient elastography to identify and follow up patients for the progression of hepatic disease.

Background: Arterial stiffness is an important determinant of cardiovascular risk. It is associated with several cardiovascular risk factors, including hypertension, diabetes and cigarette smoking. However, there are conflicting data about the relationship between arterial stiffness and hypercholesterolemia. Furthermore, augmentation index (AIx), a measure of systemic arterial stiffness, has not been previously investigated in hypercholesterolemic (HCh) children. Aim of our study was to evaluate local and systemic arterial stiffness as well as carotid intima-media thickness (IMT) in HCh children and also to investigate the relation between serum cholesterol levels and arterial stiffness., Materials and Methods: We determined lipid profile, body mass index, blood pressure, heart rate, carotid IMT and several arterial stiffness parameters, as beta-index, elastic modulus (E(p)), arterial compliance (AC), pulse wave velocity (PWV) and AIx, in 44 untreated HCh children (mean age 10.7 +/- 2.8 years; 18 with familial hypercholesterolemia, FH, and 26 with primary hypercholesterolemia, PHC) and 18 age- and sex-matched controls. HCh children never received any medication, including antihypertensive and lipid lowering drugs., Results: Respect to controls and to PHC, FH had significantly higher (P < 0.001) beta-index (5.22 +/- 1.13 vs. 3.13 +/- 0.74 and 3.60 +/- 1.02), PWV (4.72 +/- 0.72 m s(-1) vs. 3.66 +/- 0.55 m s(-1) and 4.10 +/- 0.67 m s(-1)), AIx (3.55 +/- 3.97% vs. -4.43 +/- 4.09% and 0.61 +/- 2.39%) and E(p) (64.4 +/- 19.6 kPa vs. 36.2 +/- 11.3 kPa and 42.9 +/- 13.1), whereas AC (1.25 +/- 0.48 mm(2) kPa(-1) vs. 1.9 +/- 0.43 mm(2) kPa(-1) and 1.62 +/- 0.43 mm(2) kPa(-1)) was lower (P < 0.001). There was no significant difference in carotid IMT and blood pressure values between the groups. The multiple regression analysis showed a significant association of arterial stiffness values with plasma cholesterol levels (P < 0.0001)., Conclusion: Our findings show that local and systemic arterial stiffness are increased in asymptomatic, normotensive HCh children, suggesting that HCh plays a key role in arterial mechanical impairment since the paediatric age.

Background: Biglycan (BGN), an extracellular matrix proteoglycan, has been shown to convey pro-inflammatory signals. In the present study we investigated BGN expression and its correlation with plasma levels of inflammatory markers in hypertensive subjects with or without alteration of carotid intima media thickness (IMT)., Methods: We evaluated 123 untreated essential hypertensives with no additional risk factors for atherosclerosis nor signs of cardiovascular disease and 40 controls. Hypertensives were classified according to a normal (< or =1 mm) or increased (>1 mm) IMT. BGN-mRNA and protein expression were measured in unstimulated, LPS- and Angiotensin II (Ang-II)-stimulated blood monocytes. Plasma concentrations of interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-alpha) and high sensitivity-C-reactive protein (hs-CRP) were also measured., Results: We found increased levels of IL-6, TNF-alpha, hs-CRP, and BGN-mRNA and protein in hypertensives vs controls (1.72+/-0.60 vs 1 n-fold, and 3.60+/-0.75 vs 1 n-fold, both p<0.001). However, BGN expression was not significantly different between hypertensives with IMT < or =1 mm and >1 mm. Furthermore, in vitro addition of Ang II enhanced basal BGN-mRNA (in hypertensives: 3.57+/-1.08 vs 1.72+/-0.60 n-fold, p<0.001) and protein (in hypertensives: 4.92+/-0.42 vs 3.41+/-0.75, p<0.001) expression in monocytes., Conclusions: Our data provide evidence of an enhanced expression of BGN in essential hypertension. In addition we suggest that Ang II can mediate monocyte BGN production.

Background: People with hypertension display an inflammatory pattern that includes increased plasma concentrations of monocyte chemoattractant protein 1 (MCP-1) and C-reactive protein (CRP) and enhanced expression of tissue factor (TF) mRNA in blood monocytes., Methods: In this study, we investigated the relationship between CRP concentrations and TF and MCP-1 mRNA expression in unstimulated and lipopolysaccharide (LPS)-stimulated monocytes isolated from hypertensives with or without an increase in carotid intima-media thickness (IMT). We also investigated the expression of TF and MCP-1 mRNA and MCP-1 protein after in vitro addition of CRP to monocytes. We measured CRP (by immunonephelometry) and monocyte expression of TF and MCP-1 (by real-time PCR) in 80 untreated hypertensive patients without clinical cardiovascular disease (CVD) or additional risk factors for CVD compared with 41 controls. Based on IMT measured by carotid Doppler ultrasonography, patients were classified into the categories of normal (< or =1 mm) or abnormal (>1 mm). TF and MCP-1 mRNA and MCP-1 protein (by Western blotting) were measured after in vitro addition of CRP to monocytes from 10 randomized controls as well as 10 hypertensives with IMT < or =1 mm and 10 with IMT >1 mm., Results: CRP and TF and MCP-1 mRNA concentrations were significantly higher in IMT >1 mm hypertensives vs those with IMT < or =1 mm and controls. CRP had no effect on monocyte TF mRNA from either hypertensives or controls. CRP-stimulated monocytes from hypertensives, however, showed increased MCP-1 mRNA and protein expression compared with controls and LPS-stimulated cells., Conclusions: Our findings suggest that the inflammatory response of blood monocytes plays an important role in the development of atherosclerosis and hypertension.

Background: Serum paraoxonase (PON1), a high density lipoprotein (HDL)-bound antioxidant enzyme, plays a role in atherosclerosis. An increase in PON1 activity has been reported following statin treatment., Objective: In the present study the following factors were evaluated: the influence of PON1 gene Q192R, L55M and T(-107)C polymorphisms on the response of LDL oxidisability and PON1 activity to atorvastatin treatment., Research Design and Methods: 205 Sicilian subjects with primary hypercholesterolaemia (HCh) and 69 healthy subjects as controls were concurrently enrolled. Hypercholesterolaemic patients were randomly divided into two groups: an atorvastatin group (10 mg/day atorvastatin) and a placebo group. Lipid profile, markers of LDL resistance to in vitro oxidation (lag-phase, oxidation rate and thiobarbituric acid-reactive substances), vitamin E content in LDL, PON1 activity and genotypes in both HCh and control subjects were determined at baseline. The same parameters were measured again after 3 weeks of treatment in both the atorvastatin and placebo groups., Results: HCh subjects showed significantly lower LDL resistance to oxidation, vitamin E content and PON1 activity levels than controls. A strong association was found among PON1 T(-107)C genotypes, LDL susceptibility to oxidation, vitamin E content and PON1 activity. After treatment, the atorvastatin group displayed a significant decrease in total cholesterol, LDL-cholesterol levels, and LDL susceptibility to oxidation, and an increase in vitamin E content and PON1 activity, compared with baseline values. Unlike PON1 activity levels, no difference among PON1 gene polymorphisms and reduction in markers of LDL oxidisability was observed., Conclusions: These results show, for the first time, that atorvastatin is able to improve the resistance to LDL oxidation independently of PON1 gene polymorphism.

Background: Atherosclerosis is a complex, chronic disease that usually arises from the converging action of several pathogenic processes, including hypertension, hyperlipidemia, obesity, and the accumulation of oxidized LDL. Platelet-activating factor acetylhydrolase (PAF-AH) is a LDL- and HDL-bound enzyme that hydrolyzes and inactivates PAF and prevents LDL-cholesterol oxidation, thus delaying the onset of atherosclerotic disease., Methods: We evaluated the relationship between variants of the PAF-AH gene polymorphisms Arg92His, Ile198Thr, and Ala379Val and the presence of carotid atherosclerosis in 190 hypercholesterolemic Sicilian individuals. Carotid artery intima-media wall thickness (IMT) was measured as an indicator of early atherosclerotic disease. The participants were classified according to having normal (< or =1 mm) or abnormal (> or =1 mm) IMT and were also investigated for physical characteristics and biochemical indices, including PAF-AH activity., Results: PAF-AH activity and LDL concentrations were significantly correlated in hypercholesterolemic patients, but plasma PAF-AH activity and HDL were not significantly correlated in either IMT group. No significant differences were detected among the PAF-AH gene polymorphisms in both groups after correction for age, sex, body mass index, plasma glucose and lipid concentrations, PAF-AH activity, blood pressure, and smoking habits. The analysis of PAF-AH genotype distribution showed no significant differences in percentage of 92, 198, and 379 genotypes in both IMT groups., Conclusion: Our data provided no evidence that PAF-AH polymorphisms influence PAF-AH activity and atherosclerosis in hypercholesterolemic Sicilian patients.

Age is associated with an enhanced low density lipoprotein (LDL) oxidation and atherosclerosis, thus, subjects over 80 years without cardiovascular disease provide a model to investigate the protective factors against atherosclerosis. Serum paraoxonase (PON1), an high density lipoprotein (HDL)-bound enzyme, prevents LDL oxidation. The aim of the present study was to evaluate the contribution of the PON1 promoter T(-107)C and coding region Gln192Arg (Q192R) and Leu55Met (L55M) polymorphisms to the resistance to develop cardiovascular events in Sicilian healthy octogenarians. Distribution of PON1 genotypes and activity, and biochemical parameters, were compared between 100 octogenarians and 200 adults. Individuals in the elderly group displayed significant higher levels of HDL-C (P < 0.001) and PON1 activity (P < 0.001). The analysis of PON1 genotypes distribution showed an higher percentage of (-107)CC among octogenarians compared with controls. A significant difference among T(-107)C genotypes respect to PON1 activity and HDL-C levels occurred in both groups. The CC genotype was associated with higher PON1 activity and HDL levels compared to the TT genotypes. In conclusion, our results provide a strong evidence that in healthy Sicilians ageing may be characterized by a low frequency of PON1 (-107)T 'risk' allele and by an high frequency of favourable genotypes such as (-107)CC, influencing PON1 activity and HDL-C levels.

Background and Objectives: Increased plasma low-density lipoprotein-cholesterol (LDL-C) levels in hypercholesterolemic subjects are associated with enhanced LDL oxidation that represents an additional risk for atherosclerotic disease. Human serum paraoxonase (PON1), a high-density lipoprotein (HDL) associated enzyme, has been shown to protect LDL from oxidation, thus playing an important role in reducing the risk of atherosclerosis. PON1 gene polymorphisms have been found to be associated with the variations in serum PON1 levels and activities, and with the risk for coronary artery disease (CAD). This study was performed to evaluate the contribution of the PON1 promoter (-107)T>C and the coding region Gln 192 Arg (Q192R) and Leu 55 Met (L55M) polymorphisms to the presence of carotid atherosclerosis in 208 Sicilian subjects with primary hypercholesterolemia., Methods: Carotid artery intima-media wall thickness (IMT) was measured as an indicator of early atherosclerotic disease. The subjects were classified according to whether they have a normal (1 mm) IMT. Subjects were also investigated for physical and biochemical parameters, including PON1 activity., Results: No significant differences were detected among the PON1 genotypes with respect to age, sex, BMI, plasma lipids, systolic blood pressure in both groups of patients. There were significant differences between PON1 genotypes with respect to PON1 activity. The 192QQ, 55MM and (-107)TT genotypes showed lower PON1 activity compared to the RR, LL and CC genotypes. The PON1 (-107)T>C genotype distribution in both IMT groups showed no significant differences in percentage of TT, CT and CC genotypes. Similar results were obtained analyzing the Q192R and L55M genotype frequencies. Stepwise forward logistic regression analysis confirmed the lack of association between PON1 genotypes and carotid abnormalities., Conclusions: In conclusion, our data provided no evidence of a significant association between either PON1 promoter (-107)T>C or coding region, Q192R and L55M, polymorphisms and early carotid atherosclerosis in Sicilian hypercholesterolemic subjects.

This study investigated the behavior of soluble intercellular adhesion molecule-1 (sICAM-1) and serum nitric oxide (NO) products, nitrite/nitrate (NO2-/NO3-), in subjects with primary hypercholesterolemia (HCh) without other risk factors and atherosclerosis. The effect of a short-term cholesterol-lowering treatment with atorvastatin, an HMG-CoA reductase inhibitor, on the levels of sICAM-1 and NO2-/NO3- were also investigated. After 4 weeks of placebo administration, 40 HCh (15 males and 25 females) were randomized in 2 groups: 20 subjects (atorvastatin group) received 10 mg/day of atorvastatin and the remaining 20 (placebo group) continued to take placebo. At baseline and after 4 and 12 weeks of atorvastatin or placebo administration, serum sICAM-1 and NO2-/NO3-levels were evaluated. The basal levels of these parameters were compared with those of 20 healthy subjects (C), matched for sex and age. Hypercholesterolemic subjects showed sICAM-1 and NO2-/NO3- basal values that were higher (331.7 +/- 60.3 ng/mL vs. 202.3 +/- 32.3 ng/mL, p<0.001) and lower (10.4 +/- 2.5 micromol/L vs. 20.7 +/- 4.4 micromol/L, p<0.01) than controls. No correlation between sICAM-1 or NO products and plasma cholesterol values was found, whereas there was an inverse correlation between sICAM-1 and NO2-/NO3- levels. Atorvastatin administration significantly decreased sICAM-1 and increased NO2-/NO3- levels, however these changes were not correlated with the reduction of plasma cholesterol. These data support the hypothesize that patients with HCh with no signs of arterial lesions, may have latent atherosclerosis, expressed as an increase of sICAM-1 and decrease in NO product levels. An improvement in the levels of these parameters after a short-time treatment with atorvastatin was also demonstrated.

This study was performed to determine whether the levels of soluble intercellular adhesion molecule-1 (sICAM-l) and soluble endothelial molecule-1 (sE-selectin) were elevated in subjects with hypercholesterolemia who presented with no other risk factors or evidence of atherosclerosis. The effects of administration of an HMG-CoA reductase inhibitor on the serum levels of these molecules were also examined. Forty hypercholesterolemic subjects (HCh) (19 males and 21 females), without hypertension or cardiovascular disease, received placebo for 4 weeks. The patients were then randomized in two groups; 20 of them (simvastatin group) were treated with simvastatin (20 mg/day) and the other 20 (placebo group) continued placebo administration. After 12 and 24 weeks of either simvastatin or placebo treatment, sICAM-1 and sE-selectin levels were measured. The same parameters were measured in 20 control subjects (C) with normal cholesterol levels, matched for sex and age. HCh had sICAM-1 basal values higher than C (352.4+/-57.9 ng/ml versus 114.9+/-89.6 ng/ml; P<0.001); however, sE-selectin basal values were not different in the two groups. No correlation was observed between HCh sICAM-1 levels and cholesterol levels (total and low-density lipoprotein). Furthermore, cholesterol-lowering treatment with simvastatin did not significantly diminish sICAM-1 levels. Our findings would support the hypothesis that patients with isolated hypercholesterolemia and without clinical atherosclerosis may be silent carriers of arterial subendothelial inflammation, expressed as an increase of sICAM-1.

We analysed DNA extracted from liver biopsy specimens and serum samples from 42 HCV-RNA-positive/HBsAg-negative subjects with chronic hepatitis. Twenty-eight of them were anti-HBs/anti-HBc-positive (group A), while 14 were negative for all HBV markers (group B). HBV sequences were found in hepatic DNA of 12 cases (11 of group A, one of group B), but in the serum of only two cases of group A. Sequencing analysis of pre-core region of HBV-DNA showed the presence of wild-type HBV in three cases, HBeAg-defective HBV in three cases, and the coexistence of both viral populations in six cases. These results indicate that HBV and HCV infection may coexist in HBsAg-negative chronic hepatitis, particularly in anti-HBs/anti-HBc-positive patients. However, HBV replication appears suppressed in these cases, and this state of latency may involve both wild and HBeAg-defective HBV types.

We examined nine chronic healthy hepatitis B surface antigen/antibody to hepatitis Be carriers with consistently normal liver chemistries and negative serum hepatitis B virus-DNA. Liver biopsy, performed twice, 10-11 years apart in all patients, showed normal histology and negative hepatitis B core antigen. DNA extracted from the second liver biopsy specimen, from 1 ml of serum from each patient and from an additional serum sample of 6 ml from two patients, was tested for pre-C/C and pre-S regions of hepatitis B virus-DNA by polymerase chain reaction amplification. Viral sequences were found in six of nine liver DNA extracts. In four cases both pre-C/C and pre-S regions were amplified, while the pre-C/C alone and the pre-S alone were detected in one case each. Direct sequencing of the amplified DNAs revealed no significant genomic changes in the pre-S and Core regions, while analysis of the pre-Core demonstrated the presence of a double viral population (wild-type and "e-defective") in four cases, and only "e-defective" hepatitis B virus in one case. No hepatitis B virus genomes were revealed in the serum sample when DNA was extracted from 1 ml of serum, while viral sequences were detected in both extracts of 6 ml of serum, indicating the presence of very low levels of viremia. These data suggest that episomal hepatitis B virus-DNA may persist for years in the liver of chronic healthy carriers in a latent state which may involve both wild-type and HBeAg-defective hepatitis B virus.

We studied the relations between genetic heterogeneity of pre-C region of hepatitis B virus (HBV) DNA and outcome of HBV infection in 5 infants with perinatal infection, 3 born to anti-hepatitis B e antigen (HBeAg), and 2 to HBeAg positive mothers. HBV infection developed in the babies at 3-4 months of age, but it resolved with seroconversion to anti-HBs in infants born to anti-HBe positive mothers, while the infection became chronic in the 2 babies born to HBeAg positive mothers. HBV-DNA extracted from the first hepatitis B surface antigen (HBsAg) positive serum sample of each baby was amplified and directly sequenced for the pre-core region. HBV-DNA sequences from 3 babies born to anti-HBe positive mothers showed at position 1896 the contemporary presence of 2 nucleotides (G+A), indicating a mixture of wild-type and "e minus" variant HBV. These findings suggest a possible co-transmission of the 2 viruses from anti-HBe positive mothers to newborns. HBV-DNA from babies born to HBeAg positive mothers showed wild-type sequences only. The results of this study suggest that the outcome of HBV infection in newborns depends not only on the host's immunocompetence and on viremia level in maternal blood, but also on heterogeneity of HBV. Transmission of mixed HBV populations appears associated with an early immunoelimination of the virus, while infection with wild-type HBV alone contributes to induction of chronicity.

We analyzed hepatitis B virus (HBV) genomes obtained from serum samples and liver biopsy specimen of a chronic HBsAg/anti-HBe carrier with hepatocellular carcinoma (HCC). Before the liver biopsy, performed at the time of HCC diagnosis, the patient had been followed for 2 years; the serum samples collected in that period resulted negative for HBV-DNA dot blot hybridization. The hepatic DNA was at first examined by Southern blot, but no HBV sequence was detected. Polymerase chain reaction (PCR) amplification revealed the presence of HBV genomes in DNA extracted from the liver tissue and from two serum samples collected, respectively, 1 and 2 years before the biopsy. Direct sequence of the amplified preC/C and preS regions showed that the viral populations present in serum and liver were identical and that they had a 34 nucleotide deletion in the preS2 region, while the preC region presented two mutations each introducing a translational stop codon, one at the carboxy terminal end and the other at the second codon of the region, both able to prevent HBeAg expression. These results identify a new HBV variant which was selected during a chronic infection, and had very low levels of replication as shown by its detection only after PCR amplification.

The presence of HBV-DNA sequences was evaluated in DNA extracted from serum samples, peripheral blood lymphocytes and liver biopsy specimens of five HBsAg/anti-HBe-positive carriers with chronic HDV infection. DNAs were tested by polymerase chain reaction (PCR) amplification technique using two pairs of oligonucleotide primers specific for the preC/C and S regions of the HBV; viral sequences were found exclusively in liver extracts and only in three out of the five cases. The direct sequencing of the amplified preC/C regions showed wild-type sequences in two cases, while in the third case a combination of 'wild' and 'e minus variant' viral populations was observed. Moreover, liver DNA of one positive case was electrophoresed through a low melting agarose gel and the following amplification, performed on DNA re-extracted from three different fragments of the gel, showed the presence of free HBV genomes but the absence of replicative intermediate forms. These results show that anti-HBe positivity is not constantly related to precore mutant HBV infection and suggest that HDV inhibits HBeAg production. Moreover, as it was observed in 'e minus' HBV variants, also during a chronic HBV wild-type infection, the viral replication might be suppressed to undetectable levels.