Immunometabolic and Vascular Health Responses among High Endurance Trained Subjects.

In this study, we aimed to examine the impact of high endurance training on vascular health parameters and immune-endocrine responses against modified low-density lipoprotein (LDL) particles. This observational, cross-sectional study included high endurance-trained and healthy non-trained subjects. Vascular ultrasound was used to assess vascular health parameters based on carotid intima-media thickness and endothelial function (flow-mediated dilation). Enzyme-linked immunosorbent assays were used to measure interleukin (IL)-8 and IL-10, autoantibody isotypes anti-oxidized LDL (oxLDL) and anti-apolipoprotein B (ApoB-D) peptide. Plasma levels of the corticosterone and 17 α-hydroxyprogesterone hormones were analyzed by mass spectrometry. This study enrolled 96 subjects, of whom 44 were high endurance trained and 52 were healthy non-trained individuals. Smaller carotid intima-media thickness values were observed in the high-endurance trained than in the healthy non-trained males, while no differences were observed between female groups. Flow-mediated dilation measurements did not differ by training or sex. The humoral immune responses to IgG anti-oxLDL and IgM anti-ApoB-D autoantibodies showed an isotype imbalance between the high-endurance trained and the non-trained groups. Immunoendocrine parameters showed inverse correlations between 17 α-hydroxyprogesterone concentrations and carotid intima-media thickness measurements. Direct correlations were found between IL-10 concentrations and flow-mediated dilation measurements. Chronic high-endurance exercise modulates immune-endocrine and vascular health parameters, in a sex-dependent manner.

Occupational exposure of foundry workers assessed by the urinary concentrations of 18 elements and arsenic species.

BACKGROUND: Some raw materials applied in Fe foundry industries may contain potentially toxic elements. Thus, foundry worker's occupational exposure is a constant health concern. METHOD: In this study, 194 urine samples from foundry workers were analyzed by inductively coupled plasma mass spectrometry for biomonitoring of Al, As, Ba, Cd, Co, Cr, Cs, Cu, Fe, I, Mn, Ni, Pb, Sb, Sn, Se, U and Zn. Moreover, arsenic speciation was performed in representative samples of production sector workers (group A) and administration sector workers (group B). RESULTS: Concentrations of As, Pb, Cd, Cu, Cs, I, Sb in urines from group A were higher than those found for group B. Samples of group A presented Cs, Ni, Mn, Pb, U and Zn concentrations higher than values reported for exposed workers assessed by other studies. Forty-four samples from group A exceed As-reference limits. Group A had approximately seven times more inorganic As (as arsenite) and 14 times more organic As (as dimethyl As) than group B. A statistically significant difference was observed in the elemental concentration in the workers' urine by the time in the function. Moreover, alcohol consumption is probably influencing the urine concentration of As, Ba, Cd, Co, Cu, Fe, I, Se and Zn. CONCLUSION: The monitored foundry workers are exposed to potentially toxic elements and more attention must be given to their health. Therefore, workplace safety conditions must be improved, and constant biomonitoring is necessary to ensure workers' health.

The effects of perceived chronic stress on the fMRI correlates of attentional control in women managers.

The aims of the current study were to examine differences in brain responses to cognitive control in stressed and non-stressed women managers. Stress complaints are highly prevalent among active workers and play an important role in women managers mental health and cognitive functioning. Psychosocial stress has been associated with differential executive functions in this population, but little is known about the neural correlates underlying such differences. We examined brain responses of a particular group of workers that has been proposed to have a different response to stress as a function of their status (i.e., managers). Stressed (n = 19) and non-stressed women managers (n = 21) were scanned using functional magnetic resonance imaging (fMRI) during the performance of a cognitively demanding task. We used the Stroop color-word task to compare neural activation associated with the suppression of a predominant response tendency (i.e., word reading) and the initiation of an appropriate behavior alternative (i.e., naming word color). Despite similar behavioral performances, stressed managers exhibited increased activation in the occipital cortex during response inhibition. No regions were more activated in the non-stressed relative to the stressed group. This finding of greater activation has been interpreted as compensatory brain response to maintain performance in front of cognitive challenge.

Metabolic studies of a patient harbouring a novel S487L mutation in the catalytic subunit of AMPK.

AMP-activated protein kinase (AMPK) regulates many different metabolic pathways in eukaryote cells including mitochondria biogenesis and energy homeostasis. Here we identify a patient with hypotonia, weakness, delayed milestones and neurological impairment since birth harbouring a novel homozygous mutation in the AMPK catalytic α-subunit 1, encoded by the PRKAA1 gene. The homozygous mutation p.S487L in isoform 1 present in the patient is in a cryptic residue for AMPK activity. In the present study, we performed the characterization of mitochondrial respiratory properties of the patient, in comparison to healthy controls, through the culture of skin fibroblasts in order to understand some of the cellular consequences of the PRKAA1 mutation. In these assays, mitochondrial respiratory complex I showed lower activity, which was followed by a decrement in the mtDNA copy number, which is a probable consequence of the lower expression of PGC-1α and PRKAA1 itself as measured in our quantitative PCRs experiments. Confirming the effect of the patient mutation in respiration, transfection of patient fibroblasts with wild type PRKAA1 partially restore complex I level. The preliminary clinic evaluations of the patient suggested a metabolic defect related to the mitochondrial respiratory function, therefore treatment with CoQ10 supplementation dose started four years ago and a clear improvement in motor skills and strength has been achieved with this treatment.

Determination of rivaroxaban in patient's plasma samples by anti-Xa chromogenic test associated to High Performance Liquid Chromatography tandem Mass Spectrometry (HPLC-MS/MS).

Rivaroxaban is an oral direct factor Xa inhibitor, therapeutically indicated in the treatment of thromboembolic diseases. As other new oral anticoagulants, routine monitoring of rivaroxaban is not necessary, but important in some clinical circumstances. In our study a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was validated to measure rivaroxaban plasmatic concentration. Our method used a simple sample preparation, protein precipitation, and a fast chromatographic run. It was developed a precise and accurate method, with a linear range from 2 to 500 ng/mL, and a lower limit of quantification of 4 pg on column. The new method was compared to a reference method (anti-factor Xa activity) and both presented a good correlation (r = 0.98, p < 0.001). In addition, we validated hemolytic, icteric or lipemic plasma samples for rivaroxaban measurement by HPLC-MS/MS without interferences. The chromogenic and HPLC-MS/MS methods were highly correlated and should be used as clinical tools for drug monitoring. The method was applied successfully in a group of 49 real-life patients, which allowed an accurate determination of rivaroxaban in peak and trough levels.

Detecting alcohol and illicit drugs in oral fluid samples collected from truck drivers in the state of São Paulo, Brazil.

OBJECTIVE: Alcohol and drug use by truck drivers is a current problem in Brazil. Though there is evidence that alcohol consumption is occurring in higher proportions, the use of stimulant drugs to avoid fatigue and to maintain the work schedule has also been reported. The purpose of this study was to estimate the incidence of alcohol and illicit drug use among truck drivers on São Paulo state roads. São Paulo is the most populous state in Brazil and has the largest industrial park and economic production in the country. METHODS: Data were assessed not only using a questionnaire but also, and more reliably, through toxicological analysis of oral fluid samples. Between the years 2002 and 2008, 1250 oral fluid samples were collected from truck drivers on the roads during morning hours. The samples were tested for the presence of alcohol, cocaine, tetrahydrocannabinol (THC), and amphetamine/methamphetamine. A previously published, validated gas chromatographic (gas chromatography-flame ionization detection and gas chromatography-mass spectrometry) method was applied to the samples for alcohol and drug detection. RESULTS: Of the total analyzed samples, 3.1 percent (n = 39) were positive: 1.44 percent (n = 18) were positive for alcohol, 0.64 percent (n = 8) for amphetamines, 0.56 percent (n = 7) for cocaine, and 0.40 percent (n = 5) for THC. In one case, cocaine and THC were detected. The results are indicative of the extent of alcohol and drug use by truck drivers in the state of São Paulo, Brazil. CONCLUSIONS: This research provides evidence that not only alcohol but also illicit drug use is a real problem among professional drivers. The use of these substances should be controlled to better promote safe driving conditions on Brazilian roads.

Determination of a method for extraction of coenzyme Q10 in human plasma: optimization of the use of surfactants and other variables.

OBJECTIVE: To establish a routine for the extraction of the total levels of CoQ10 in human plasma through the Ultra High Performance Liquid Chromatography (UHPLC). METHODS: Two extraction protocols were tested: a) methanol: hexane and b) 1-propanol. The following parameters were analyzed: extraction temperature (19ºC and 4ºC), extraction tubes (glass and polypropylene), and surfactants (SDS, Triton X-100, Tween-20) at different concentrations, i.e., 1%, 3%, 5% and 10%. RESULTS: The results showed that the method of extraction of CoQ10 in a sample of human plasma at 4ºC, using solvents methanol: hexane (85:15, v/v) in the presence of surfactant Tween-20 at 3% and polypropylene tubes showed better efficiency and reproducibility when compared to the method with 1-propanol. CONCLUSION: By the analyses performed, it was possible to observe that the addition of the surfactant Tween-20 promoted an increase in the recovery of CoQ10 by the methanol:hexane extraction method. This method showed good reproducibility, with a low coefficient of variation and high sensitivity, since CoQ10 was detected in samples of plasma of a control individual using a UV-type detector. The use of UHPLC equipment allowed a total analysis with total run time of 3.5 minutes, enabling the rapid achievement of results, considered mandatory for laboratory routines.

Determination of a method for extraction of coenzyme Q10 in human plasma: optimization of the use of surfactants and other variables / Determinação de um método de extração de coenzima Q10 em plasma humano: otimização do uso de surfactantes e outras variáveis

Objective: To establish a routine for the extraction of the total levels of CoQ10 in human plasma through the Ultra High Performance Liquid Chromatography (UHPLC). Methods: Two extraction protocols were tested: a) methanol: hexane and b) 1-propanol. The following parameters were analyzed: extraction temperature (19ºC and 4ºC), extraction tubes (glass and polypropylene), and surfactants (SDS, Triton X-100, Tween-20) at different concentrations, i.e., 1%, 3%, 5% and 10%. Results: The results showed that the method of extraction of CoQ10 in a sample of human plasma at 4ºC, using solvents methanol: hexane (85:15, v/v) in the presence of surfactant Tween-20 at 3% and polypropylene tubes showed better efficiency and reproducibility when compared to the method with 1-propanol. Conclusion: By the analyses performed, it was possible to observe that the addition of the surfactant Tween-20 promoted an increase in the recovery of CoQ10 by the methanol:hexane extraction method. This method showed good reproducibility, with a low coefficient of variation and high sensitivity, since CoQ10 was detected in samples of plasma of a control individual using a UV-type detector. The use of UHPLC equipment allowed a total analysis with total run time of 3.5 minutes, enabling the rapid achievement of results, considered mandatory for laboratory routines.

Objetivo: Estabelecer uma rotina de extração dos níveis totais de CoQ10 em plasma humano por meio da análise por Cromatografia Líquida de Ultra Alta Eficiência (UHPLC). Métodos: foram testados dois protocolos de extração: a) metanol:hexano e b) 1-propanol. Os seguintes parâmetros foram analisados: temperatura de extração (19ºC e 4ºC), tubos de extração (vidro e polipropileno), surfactantes (SDS, Triton X-100, Tween-20) em diferentes concentrações 1%, 3%, 5% e 10%. Resultados: Os resultados mostraram que o método de extração de CoQ10 em amostra de plasma humano, a 4ºC, utilizando-se os solventes metanol:hexano (85:15, v/v) na presença do surfactante Tween-20 a 3% e tubos de polipropileno apresentou melhor eficiência e reprodutibilidade quando comparado ao método com 1-propanol. Conclusão: A adição do surfactante Tween-20 no processo de preparação de amostra promoveu um aumento na recuperação da CoQ10 pelo método de extração metanol:hexano observada pela boa reprodutibilidade das prelicatas, pelo baixo coeficiente de variação e alta sensibilidade uma vez que a CoQ10 foi detectada em amostras de plasma de um indivíduo controle utilizando-se um detector do tipo UV. Além disso, a utilização de um equipamento de UHPLC proporcionou a obtenção de uma análise com tempo total de corrida de 3,5 minutos, o que viabiliza a obtenção rápida de resultados, considerado mandatório para rotinas laboratoriais.

Determination of opiates in whole blood and vitreous humor: a study of the matrix effect and an experimental design to optimize conditions for the enzymatic hydrolysis of glucuronides.

Undoubtedly, whole blood and vitreous humor have been biological samples of great importance in forensic toxicology. The determination of opiates and their metabolites has been essential for better interpretation of toxicological findings. This report describes the application of experimental design and response surface methodology to optimize conditions for enzymatic hydrolysis of morphine-3-glucuronide and morphine-6-glucuronide. The analytes (free morphine, 6-acetylmorphine and codeine) were extracted from the samples using solid-phase extraction on mixed-mode cartridges, followed by derivatization to their trimethylsilyl derivatives. The extracts were analysed by gas chromatography-mass spectrometry with electron ionization and full scan mode. The method was validated for both specimens (whole blood and vitreous humor). A significant matrix effect was found by applying the F-test. Different recovery values were also found (82% on average for whole blood and 100% on average for vitreous humor). The calibration curves were linear for all analytes in the concentration range of 10-1,500 ng/mL. The limits of detection ranged from 2.0 to 5.0 ng/mL. The method was applied to a case in which a victim presented with a previous history of opiate use.