The effect of sex/gender on cardiovascular pharmacology.

Cardiovascular diseases differ between men and women as do outcomes after therapeutic interventions. Management of these diseases, however, is generally guided by evidence from trials conducted predominantly in men, with few studies focused on women alone. Our goal is to review the sex/gender differences in cardiovascular therapies, which show many areas of uncertainty regarding women due to their small enrolment in clinical trials; thus, in some cases, firm conclusions about efficacy in women are difficult to obtain.

Carotid restenosis is associated with plasma ADMA concentrations in carotid endarterectomy patients.

Background: The aim of this work has been to study the association between plasma asymmetric dimethylarginine (ADMA) concentrations and carotid stenosis in a group of 64 patients undergoing carotid endarterectomy (CEA).

Methods: Arginine, ADMA and symmetric dimethylarginine (SDMA) were measured using capillary electrophoresis with UV detection. An evaluation of plasma concentrations of total cysteine (tCys) and total homocysteine (tHcy) was also performed.

Field-amplified sample injection combined with pressure-assisted capillary electrophoresis UV detection for the simultaneous analysis of allantoin, uric acid, and malondialdehyde in human plasma.

The allantoin/uric acid (All/UA) ratio and malondialdehyde (MDA) plasma levels have been proposed as important markers for monitoring oxidation triggered by the action of free radicals (FR). Here, we describe an easy field amplified sample injection capillary electrophoresis method with UV detection for the separation and quantification of All, UA, and free MDA in human plasma. The plasma samples were simply filtered through centrifugation membrane tubes for protein elimination and directly injected on a capillary without complex cleanup and/or sample derivatization procedures.

Improved method for plasma ADMA, SDMA, and arginine quantification by field-amplified sample injection capillary electrophoresis UV detection.

Here, we describe an easy field-amplified sample injection capillary electrophoresis method with UV detection for the separation and detection of free plasma arginine and dimethylated arginines. The analytes were baseline-separated within 22 min by using 50 mmol/L Tris phosphate pH 2.3 as running buffer.

Evaluation of methylation degree from formalin-fixed paraffin-embedded DNA extract by field-amplified sample injection capillary electrophoresis with UV detection.

Alterations in global DNA methylation are implicated in several pathobiological processes. The tissues stored as paraffin blocks represent an important source of DNA for retrospective genetic and epigenetic analysis on a large scale. Therefore, we developed the first capillary electrophoresis method able to measure global methylation in formalin-fixed, paraffin-embedded (FFPE) DNA extracts.

High sensitivity of reverse-hybridization methodology in the detection of KRAS mutations from formalin-fixed paraffin-embedded colorectal cancer samples.

Colorectal cancer is ranked the third most common cancer worldwide in terms of incidence and the second in terms of mortality. Recent advances in therapeutic approaches to colorectal cancer have identified a potential role of anti-epidermal growth factor receptor (EGFR) targeted therapies as adjuvant treatment in advanced disease. New evidences showed that patients harboring KRAS mutations on codons 12 and 13 are not responsive to anti-EGFR monoclonal antibodies.

Plasma asymmetric dimethylarginine (ADMA) levels and atherosclerotic disease in ankylosing spondylitis: a cross-sectional study.

Conclusive data about the prevalence of endothelial dysfunction and atherosclerotic process in ankylosing spondylitis (AS) patients with respect to the general population are lacking. Elevated plasma levels of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, have been reported in clinical conditions associated with endothelial dysfunction and atherosclerotic disease. We performed a cross-sectional study to evaluate plasma ADMA levels and atherosclerotic disease in AS patients.

Quantification of histidine, 1-methylhistidine and 3-methylhistidine in plasma and urine by capillary electrophoresis UV-detection.

We describe a new CE method with UV-detection for the quantification of histidine (His) and its methylated forms 1-methylhistidine and 3-methylhistidine, both in plasma and urine. Analytes were basically resolved using a 60 mmol/L Tris-phosphate run buffer pH 2.2 in less than 12 min.

Capillary electrophoresis with laser-induced fluorescence detection for ATP quantification in spermatozoa and oocytes.

We describe a new capillary electrophoresis laser-induced fluorescence (CE-LIF) method for the quantification of adenosine 5'-triphosphate (ATP) in spermatozoa and oocytes. The optimization of the precapillary derivatization reaction between ATP and 4,4-difluoro-5,7-dimethyl-4-bora-3a,4adiaza-s-indacene-3-propionyl ethylene diamine hydrochloride (BODIPY FL EDA) has been described. BODIPY-ATP conjugate was analysed in an uncoated fused silica capillary of 75 μm ID and 50 cm effective length using a 10 mmol/L tribasic sodium phosphate buffer, pH 11.

Quantification of neurotransmitter amino acids by capillary electrophoresis laser-induced fluorescence detection in biological fluids.

The role of neurotransmitter amino acids (NAAs) in the functioning of the nervous system has been the focus of increasingly intense research over the past several years. Among the various amino acids that have important roles as neurotransmitters, there are alanine (Ala), glutamic acid (Glu), aspartic acid (Asp), serine (Ser), taurine (Tau) and glycine (Gly). NAAs are present in plasma, cells and--at trace levels--in all biological fluids, but complex components in biological matrices make it difficult to determine them in biological samples.

Determination of homocysteine thiolactone, reduced homocysteine, homocystine, homocysteine-cysteine mixed disulfide, cysteine and cystine in a reaction mixture by overimposed pressure/voltage capillary electrophoresis.

An elevated level of thiol amino acid homocysteine is associated with several complex disorders. Homocysteine ability to bind proteins, thereby modulating their structure and function, is proposed to be one of the mechanisms of homocysteine induced pathogenecity. Homocysteine and homocysteine thiolactone bind to protein cysteine and lysine residues respectively.

Increased low-density lipoprotein S-homocysteinylation in chronic kidney disease.

Background: Since low-density lipoprotein (LDL) S-homocysteinylation has been recently reported to enhance atherogenicity of lipoprotein, we have investigated the levels of homocysteine (Hcy) linked to LDL in chronic proteinuric patients in which lipid abnormalities highly contribute to the excess of morbidity and mortality.

Methods: We used capillary electrophoresis to measure LDL-bound thiol Hcy, cysteine (Cys), cysteinylglycine (Cys-Gly), glutathione (GSH), and glutamylcysteine (Glu-Cys) in 30 chronic kidney disease (CKD) individuals and 60 healthy volunteers.

Results: We found more elevated levels of total plasma Hcy, Cys, GSH and Glu-Cys in patients than in controls and also found that Hcy and Cys bound to LDL were significantly increased in nephropathic subjects.

Ultra-fast adenosine 5'-triphosphate, adenosine 5'-diphosphate and adenosine 5'-monophosphate detection by pressure-assisted capillary electrophoresis UV detection.

Herein, we report a new CE method to measure adenine nucleotides adenosine 5'-triphosphate, adenosine 5'-diphosphate, and adenosine 5'-monophosphate in red blood cells. For this purpose, 20 mmol/L sodium acetate buffer at pH 3.80 was used as running electrolyte, and the separation was performed by the simultaneous application of a CE voltage of 25 kV and an overimposed pressure of 0.

Differences in semen freezability and intracellular ATP content between the rooster (Gallus gallus domesticus) and the Barbary partridge (Alectoris barbara).

This study aimed to compare viability, ATP content, and DNA integrity of rooster (Gallus gallus domesticus) and Barbary partridge (Alectoris barbara) fresh and frozen spermatozoa in order to identify factors possibly related to differences in semen freezability. Ejaculates were obtained from March to May by the abdominal massage method from 3 adult roosters and 12 adult Barbary partridges. Semen was frozen with different cryoprotectants using Lake's diluents as a base medium: 1) glycerol 11%; 2) glycerol 11% and trehalose 70 mmol/L; 3) dimethylacetamide (DMA) 6%; 4) DMA 6% and trehalose 70 mmol/L.

Plasma thiols and taurine levels in central retinal vein occlusion.

Purpose: To determine the plasma levels of the sulfur-containing amino-acids homocysteine, cysteine, cysteinylglycine, glutamylcysteine, glutathione, and taurine in patients with central retinal vein occlusion (CRVO) and in healthy subjects and to ascertain whether there are statistically significant differences between patients and controls.

Methods: Laser-induced fluorescence capillary electrophoresis was used to measure the plasma levels of homocysteine, cysteine, cysteinylglycine, glutamylcysteine, glutathione, and taurine in 29 patients with CRVO and 80 age- and gender-matched control subjects. Wilcoxon or Student's t-test was used, when appropriate, to determine differences between groups.

Sexual dimorphic evolution of metabolic programming in non-genetic non-alimentary mild metabolic syndrome model in mice depends on feed-back mechanisms integrity for pro-opiomelanocortin-derived endogenous substances.

Previously, we showed that our post-natal handling model induces pro-opiomelanocortin-derived (POMC) endogenous systems alterations in male mice at weaning. These alterations last up to adult age, and are at the basis of adult hormonal and metabolic conditions similar to mild metabolic syndrome/type-2 diabetes. Here, we evaluate how sex influences post-natal programming in these metabolic conditions.

A hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) method for genomic DNA methylation assessment by UV detection.

A hydrophilic interaction chromatography-based method, in combination with 1.7 microm ethylene bridged hybrid particle packed column (100 mm x 2.1 mm I.

Hemostatic gene polymorphisms in young Sardinian with non-fatal acute myocardial infarction.

Although the role of environmental factors in the development of acute myocardial infarction (AMI) has been clearly established, the role of genetic factors is still undefined. The aim of this study was to investigate the association between various gene polymorphisms in the haemostatic system and the risk of myocardial infarction in a very genetic restricted area population of Sardinian young adults with AMI. The study case-control involved 71 patients who had survived a first MI at a mean age of 47,2 years and 150 healthy subjects.

Albumin-bound low molecular weight thiols analysis in plasma and carotid plaques by CE.

We describe a new method for the quantification of low molecular weight thiols, as homocysteine, cysteine, cysteinylglycine, glutamylcysteine and glutathione bound to human plasma albumin. After albumin isolation and purification by SDS-PAGE, thiols were freed from protein with tri-n-butylphosphine and successively derivatized with 5-iodoacetamidofluorescein. Samples were then injected and quantified in about 18 min by CE with laser induced fluorescence detection.

Decreased plasma cysteinylglycine and taurine levels in branch retinal vein occlusion.

Purpose: Our aim was to determine the plasma levels of the sulfur-containing amino acids homocysteine, cysteine, cysteinylglycine, glutamylcysteine, glutathione and taurine in patients with branch retinal vein occlusion (BRVO) and in healthy subjects and to ascertain whether there are statistically significant differences between patients and controls.

Methods: Homocysteine, cysteine, cysteinylglycine, glutamylcysteine, glutathione and taurine plasma levels were measured in 40 patients with BRVO and 80 age- and gender-matched control subjects by using laser-induced fluorescence capillary electrophoresis methods. Wilcoxon's or Student's t test was used, when appropriate, to determine differences between the groups.

Improved rapid assay of plasma uric acid by short-end injection capillary zone electrophoresis.

A rapid and simple short-end injection capillary zone electrophoresis method was developed for the quantification of plasma uric acid. The separation was performed in an uncoated fused-silica capillary (50 microm ID, 60 cm total length, 10.2 cm effective length) by using as a background electrolyte a 75 mmol/L glycylglycine solution titrated with NaOH 5 mol/L to pH 9.

High dietary taurine reduces apoptosis and atherosclerosis in the left main coronary artery: association with reduced CCAAT/enhancer binding protein homologous protein and total plasma homocysteine but not lipidemia.

We sought to determine whether taurine could specifically protect against coronary artery disease during an atherogenic diet and whether taurine affects the lipid profile, metabolites of methionine, and endothelial atherogenic systems. Rabbits were fed one of the following diets for 4 weeks: (1) control diet; (2) 0.5% cholesterol+1.

S-homocysteinylated LDL apolipoprotein B adversely affects human endothelial cells in vitro.

Objective: In recent years elevated homocysteine (Hcy) levels have been widely recognized as a risk factor for cardiovascular diseases (CVDs) and a connection between hyperhomocysteinemia and lipid metabolism has been suggested to have a possible role in endothelial vascular damage as lipoprotein fractions contain higher Hcy levels in hypercholesterolemia, compared to normolipidemic individuals. However, the biochemical events underlying the interaction between Hcy and LDL are still poorly understood.

Methods And Results: Herein we have investigated the interaction of LDL with Hcy by measuring thiols S-linked to apoprotein using capillary electrophoresis and have evaluated the effect of S-homocysteinylated LDL on human endothelial cells (HECs).

Evaluation of low molecular mass thiols content in carotid atherosclerotic plaques.

Objectives: Despite the evidence that both homocysteine and cysteine are important risk factors for vascular disease and atherosclerosis no information are reported about their effective amount in plaque and on the relationship with the other low molecular weight thiols.

Design And Methods: We used capillary electrophoresis to measure thiols in both carotid plaque specimens and plasma samples from 37 patients undergoing carotid endarterectomy.

Results: Pearson's correlation shows that intraplaque homocysteine, cysteine and cysteinylglycine levels are related to their plasma concentrations.