Stratification of ST-elevation myocardial infarction patients based on soluble CD40L longitudinal changes.

Involvement of soluble CD40 ligand (sCD40L) in thrombosis and inflammation on the context of coronary artery disease is currently being revised. In that perspective, we had studied the association of sCD40L with markers of platelet activation and markers of endothelial and vascular function. On that cohort, a stratification of patients with acute myocardial infarction (AMI) 1 month after percutaneous coronary intervention (PCI) was observed based on concentrations of sCD40L. The study intended to identify the groups of AMI patients with different profiles of sCD40L concentrations and verify how medication, clinical evolution, biochemical data, and markers of regulation of endothelial function at genetic (endothelial nitric oxide synthase polymorphisms) and post-transcriptional levels (circulating microRNAs) affect sCD40L serum levels. Lower quartiles of sCD40L (<2.3 ng/mL) were associated with higher concentrations of N-terminal pro-brain natriuretic peptide (NT-proBNP), high frequency of G894T polymorphism, and altered expression of a set of microRNAs assumed to be involved in the regulation of endothelial and cardiac function and myocardium hypertrophy, relative to patients in sCD40L upper quartiles. A characteristic sCD40L variation pattern in STEMI patients was identified. Low levels of sCD40L 1 month after PCI distinguish STEMI patients with worse prognosis, a compromised cardiac healing, and a persistent endothelial dysfunction, as given by the association between sCD40L, NT-proBNP, G894T polymorphism, and specific profile of miRNA expression. These results suggest sCD40L could have a prognostic value in STEMI patients.

Changes of soluble CD40 ligand in the progression of acute myocardial infarction associate to endothelial nitric oxide synthase polymorphisms and vascular endothelial growth factor but not to platelet CD62P expression.

Reported in vitro data implicated soluble CD40 ligand (sCD40L) in endothelial dysfunction and angiogenesis. However, whether sCD40L could exert that influence in endothelial dysfunction and angiogenesis after injury in acute myocardial infarction (AMI) patients remains unclear. In the present study, we evaluated the association of sCD40L with markers of platelet activation, endothelial, and vascular function during a recovery period early after AMI. To achieve this goal, the time changes of soluble, platelet-bound, and microparticle-bound CD40L levels over 1 month were assessed in AMI patients and correlated with endothelial nitric oxide synthase (eNOS) polymorphisms, vascular endothelial growth factor (VEGF) concentrations, and platelet expression of P-selectin (CD62P). The association of soluble form, platelet-bound, and microparticle-bound CD40L with CD62P expression on platelets, a marker of platelet activation, was also assessed to evaluate the role of CD40L in the thrombosis, whereas the association with eNOS and VEGF was to evaluate the role of CD40L in vascular dysfunction. This work shows for the first time that time changes of sCD40L over 1 month after myocardial infarct onset were associated with G894T eNOS polymorphism and with the VEGF concentrations, but not to the platelet CD62P expression. These results indicate that, in terms of AMI pathophysiology, the sCD40L cannot be consider just as being involved in thrombosis and inflammation but also as having a relevant role in vascular and endothelial dysfunction.

Prognostic value of VEGF in patients submitted to percutaneous coronary intervention.

We examined the longitudinal changes of VEGF levels after percutaneous coronary intervention for predicting major adverse cardiac events (MACE) in coronary artery disease (CAD) patients. VEGF was measured in 94 CAD patients' serum before revascularization, 1-month and 1-year after. Independently of clinical presentation, patients had lower VEGF concentration than a cohort of healthy subjects (median, IQ: 15.9, 9.0-264 pg/mL versus 419, 212-758 pg/mL; P < 0.001) at baseline. VEGF increased to 1-month (median, IQ: 276, 167-498 pg/mL; P < 0.001) and remained steady to 1-year (median, IQ: 320, 173-497 pg/mL; P < 0.001) approaching control levels. Drug eluting stent apposition and previous medication intake produced a less steep VEGF evolution after intervention (P < 0.05). Baseline VEGF concentration <40.8 pg/mL conveyed increased risk for MACE in a 5-year follow-up. Results reflect a positive role of VEGF in recovery and support its importance in CAD prognosis.

Crosstalk between inflammation, iron metabolism and endothelial function in Behçet's disease.

Behçet's disease (BD) is a rare chronic vasculitis of unclear etiology. It has been suggested that inflammatory response has an important role in BD pathophysiology. Herein, we aimed to study the interplay between inflammation, iron metabolism and endothelial function in BD and search for its putative association with disease activity. Twenty five patients clinically diagnosed with BD were selected and twenty four healthy age-sex matched individuals participated as controls. Results showed an increase of total number of circulating white blood cells and neutrophils, serum transferrin, total iron binding capacity, mieloperoxidase (MPO), ceruloplasmin (Cp), C reactive protein, ß2 microglobulin and Cp surface expression in peripheral blood monocytes in BD patients comparatively to healthy individuals (p < 0,05). Of notice, the alterations observed were associated to disease activity status. No significant differences between the two groups were found in serum nitric oxide concentration. The results obtained suggest an important contribution from innate immunity in the pathogenesis of this disease. In particular, surface expression of leukocyte-derived Cp may constitute a new and relevant biomarker to understand BD etiology.

T lymphocytes alterations are associated with oxidized LDL, troponin T, white blood cells and C-reactive protein during acute myocardial infarction.

BACKGROUND: Auto-immune responses are associated with oxidized LDL (ox-LDL) release, a key factor in plaque destabilization. Data on the relationship between ox-LDL and T lymphocytes in human populations remains scarce. T cells also react with other molecules from the lesion and/or damage the myocardium. OBJECTIVE: The objective of the present study was to examine the relationship between circulating T lymphocytes, ox-LDL, markers of myocardial necrosis (cTnT), myocardial dysfunction (N-terminal pro-brain natriuretic peptide - NT-proBNP) and inflammation (C-reactive protein - CRP) in the setting of acute myocardial infarction. METHODS: A longitudinal study of 55 patients with ST-elevation myocardial infarction (STEMI) were evaluated at three time points: admission, 2 and 40 days following admission, together with 30 patients with stable angina (SA) and 56 subjects without coronary artery disease serving as controls (CTR). RESULTS: STEMI patients had maximal ox-LDL values and minimal levels of CD3+ T lymphocytes at admission, which was normalized during the recovery period. The increasing trend of CD3+ T cells was positively associated with an ox-LDL decline over time. CRP and cTnT longitudinal variations were negatively associated with the CD3+ T-cell increasing trend. These associations were not found in SA patients or controls. CONCLUSIONS: The associations found between CD3+ T lymphocytes, ox-LDL and cTnT suggest a specificity of the immune response in AMI towards arterial and myocardial inflammation and remodelling.

Serial changes in oxidized low-density lipoprotein associated with culprit vessel in ST-elevation myocardial infarction--a promising marker?

The aim of the present study was to investigate variations in oxidized LDL (oxLDL) at the onset of acute myocardial infarction (AMI) and over the recovery period, exploring their relationship with coronary disease severity. A follow-up of 50 AMI patients was evaluated against 25 healthy volunteers (reference group). The AMI patients were evaluated at three time points: at admission before the administration of IIb/IIIa inhibitors and angioplasty, and two and 40 days after intervention. Plasma oxLDL concentrations were measured by ELISA. oxLDL was found to be significantly higher in AMI patients in the acute phase relative to reference levels, decreasing progressively over the recovery period. The results also demonstrated that oxLDL levels were decreased in patients with the left circumflex artery (LCX) as culprit vessel compared to the left anterior descending coronary (LAD) or right coronary artery (RCA). The results highlight a significant increase in oxLDL concentration related to coronary artery disease severity, as conditions such as LCX lesions are usually associated with a favorable prognosis, contrasting with LAD-associated conditions that can compromise large areas of myocardium. The results thus suggest that oxLDL may constitute a promising marker in assessment of AMI evolution.

Influence of dyslipidemia and smoking on redox markers in humans--a critical study.

UNLABELLED: Experimental research indicates that oxidative processes play a role in susceptibility to a large number of diseases. A better understanding of the parameters affecting redox balance could delay and even prevent such processes. OBJECTIVE: The present study aims to investigate blood parameters associated with antioxidant systems in a Portuguese population for the first time, taking into consideration gender, age range, lipid profile and smoking habits as influencing factors. DESIGN AND PARTICIPANTS: One hundred and eighty-three healthy Portuguese subjects of both genders were recruited from the metropolitan area of Lisbon. The group consisted of individuals aged from 20 to 70 years, who gave their informed consent before participating in the study. All subjects were screened to determine eligibility, which was based on a clinical report. Subjects were considered eligible if they had no acute or chronic illness and were not taking any drugs or dietary supplements that could compromise the values of the studied parameters. The subjects were then divided into different subgroups according to gender, age range, lipid profile and smoking habits. METHODS: Whole blood glutathione peroxidase activity and serum albumin, transferrin and uric acid were determined using commercially available kits. Superoxide dismutase activity in erythrocytes and thiobarbituric acid reactive substances in serum were measured using methods published elsewhere. RESULTS: Glutathione peroxidase activity was not affected by any of the studied variables, but superoxide dismutase activity decreased with smoking. Albumin levels remained unchanged under all conditions. Hyperlipidemia was associated with higher lipid peroxidation as well as higher uric acid levels. Gender was the strongest predictor for transferrin, total iron binding capacity and uric acid variations. Finally, a multivariate statistical model clearly separated genders and lipid profile and genders and smoking. CONCLUSIONS: The present study suggests that hyperlipidemia and smoking should be considered important selection criteria in epidemiological studies focusing on oxidative stress and on the atherosclerotic process.

Metallothionein levels in Algerian mice (Mus spretus) exposed to elemental pollution: an ecophysiological approach.

The potential use of metallothioneins (MTs) as biomarkers of trace metal contamination was evaluated for the first time in the Algerian mouse (Mus spretus). Mice were collected seasonally in an abandoned mining area (Aljustrel) and in a reference area, both located in southern Portugal. MT levels were quantified in liver and kidney by differential pulse polarography and hepatic elemental concentrations (Mn, Fe, Cu, Zn, Se) were determined by particle-induced X-ray emission. Hepatic iron and selenium concentrations were elevated in mice from Aljustrel mine when compared to reference animals. MTs levels were averagely higher in mice from Aljustrel than those originated from the reference area. A season-dependent significant effect was found on the hepatic and renal MT concentrations, characterized by higher levels in winter and lower in autumn. In contaminated mice positive relationship between liver elemental contents (Cu in autumn and Fe in winter) and MTs were found. The seasonal variation of MT suggests that probably physiological and environmental factors could influence hepatic and renal MT induction. Results seem to imply that some environmental disturbance occur in the vicinity of the Aljustrel mine. Therefore, for the management purposes MT levels should be followed in liver of M. spretus, especially in winter. Furthermore, other physiological factors that could influence MT expression and turnover in Algerian mouse should also be monitored.

Comparison of apolipoprotein E polymorphism and other atherosclerotic risk factors in two portuguese populations: the influence of insularity

The main objective of this study was to investigate the apolipoprotein (apo) E genotypes in Portuguese populations from mainland (Lisbon city) and from San Miguel Island, Azores' Archipelago (Ponta Delgada city) and to look for differences between these particular sites in apparently healthy subjects. Also, subjects with clinical diagnosis of atherosclerotic disease were investigated in San Miguel Island. In Lisbon, the genotypes distribution was: epsilon3/epsilon 3 > epsilon 3/epsilon 4 > epsilon 2/epsilon 3 > epsilon 4/epsilon 4 while that, for Ponta Delgada and regardless the health condition, was: epsilon 3/epsilon 3 > epsilon 2/epsilon 3 > epsilon 3/epsilon 4. Within Ponta Delgada control group, females and males had distinct genotype frequencies. The most common atherosclerotic risk factors as body mass index, blood hypertension and serum lipid parameters, presented some differences among the allelic subgroups of apo E. The major conclusions were: 1) an apparent influence of insularity in apo E polymorphism was observed; 2) both the high risk genotypes epsilon 2/epsilon 2 and epsilon 2/epsilon 4 were not found, even in patients; 3) curiously, the genotypes proportion in females was not homogenous among the three groups.

O principal objectivo deste estudo é o de pesquisar o efeito da insularidade nos polimorfismos da apolipoproteína (apo) E em indivíduos saudáveis do continente (Lisboa) e de Ponta Delgada (Ilha de S. Miguel, Arquipélago dos Açores). Adicionalmente, estudar a distribuição dos seus genótipos em doentes com aterosclerose da Ilha de S. Miguel. Em Lisboa, a distribuição dos genótipos da apo E foi a seguinte: épsilon3/épsilon3 > épsilon3/épsilon4 > épsilon2/épsilon3 > épsilon4/épsilon 4, ao passo que em Ponta Delgada e independentemente da condição fisiológica foi: épsilon 3/épsilon 3 > épsilon 2/épsilon 3 > épsilon 3/épsilon 4. Distintas frequências genotípicas foram observadas entre homens e mulheres no grupo saudável de Ponta Delgada. O índice de massa corporal, hipertensão arterial e perfil lipídico, factores de risco associados ao processo aterosclerótico, revelaram algumas diferenças quando avaliados em função dos grupos alélicos. Neste estudo, os genótipos de risco da apo E, épsilon2/épsilon2 e épsilon2/épsilon4, não foram contabilizados. Curiosamente a proporção dos genótipos nas mulheres foi heterogénea nos 3 grupos estudados.

How does the greater white-toothed shrew, Crocidura russula, responds to long-term heavy metal contamination? -- A case study.

Heavy metals accumulation in parallel with the evaluation of physiological and biochemical effects resulting from continued metal exposure were considered here using for the first time the great white-toothed shrew Crocidura russula as an in vivo model. Shrews were originated from an abandoned lead/zinc mining area and from a reference area, both in Alentejo, southern Portugal. Hepatic contents of nickel, copper, zinc, cadmium, mercury and lead were quantified by Inductively Coupled Plasma Mass Spectrometer (ICP-MS). Haematological parameters (white blood cells, red blood cells, haemoglobin and haematocrit) were obtained in a Coulter Counter Analyser and biochemical markers of the redox balance (glutathione S-transferase, glutathione peroxidase, and glutathione reductase) activities were measured spectrophotometrically using a Duo-50 spectrophotometer. Compared with control animals, significantly higher concentration of hepatic cadmium (9.29 vs. 1.18 micorg/g dry weight) and nickel (1.56 vs. 0.343 microg/g dry weight) were detected in the shrews collected in the mining area. However, no significant changes were observed on haematological or enzymatic parameters in animals exposed to metal pollution. The obtained results show that shrews are good bioaccumulators of toxic heavy metals, but very tolerant to their effects, revealing an interesting long-term adaptation to polluted environments. In addition, this study provides reference values for haematological parameters and antioxidant enzymes levels in C. russula, which may be relevant for comparative purposes in further studies.

Polymorphism of the APOE locus in the Azores Islands (Portugal).

Our aim in this study is to report on the polymorphism of the APOE gene in the Azores Islands (Portugal) to obtain a population baseline of the existing variation in this locus, known to be one of the genetic determinants of plasma lipid levels. One hundred twenty-six Azorean individuals were typed for the APOE polymorphism using standard PCR-RFLP. Allele frequencies obtained for APOE*2, APOE*3, and APOE*4 were 6.75%, 83.73%, and 9.52%, respectively. The APOE*3/*3 genotype presented the highest frequency (69.84%), and the APOE*4/*4 genotype had the lowest frequency (0.79%). Genotype frequencies were in conformity with Hardy-Weinberg expectations. The observed genotype and allele frequencies were similar to those reported for other Iberian samples. Furthermore, Nei's gene diversity (H = 0.2864 +/- 0.0351) was similar to that reported for samples from mainland Portugal. The data generated from this study will be of importance in the context of ongoing studies concerning the factors that influence lipid levels in the Azorean population.