Standardization of platelet parameters determination in blood samples collected with EDTA

The Abbott Cell-Dyn 3000 automated hematological analyzer prepares a histogram of platelet volume, which is measured by a technique using electrical impedance, inferring from its platelet count (PLT), mean platelet volume (MPV) and platelet distribution width (PDW). This equipment also calculates the plateletcrit (PCT). These platelet parameters may be important to evaluate platelet function, but they require standardization, because platelets swell when in contact with ethylenediaminetetra-acetic acid salts, hence an increase in blood sample storage time produces artificially increased results. To assess the effect of storage time on MPV, PLT, PDW and PCT, blood samples from 23 sickle cell anemia patients during steady state (Group I) and 50 from healthy controls (Group II) were placed in Vacutainer® tubes with dipotassium ethylenediaminetetra-acetic acid and measured over a period of 1440 minutes (24 h) at the following times: immediately after the venipuncture (time 0), 15, 30, 60, 120, 240, 360, 480 and 1440 minutes. The mean values of MPV and PCT were significantly increased (p<0.00001) along the storage time in both groups. The mean values of PLT and PDW were practically stable (p>0.05) throughout the storage time in both groups.

An optimised electrochemical biosensor for the label-free detection of C-reactive protein in blood

C-reactive protein (CRP) is an acute phase protein whose levels are increased in many disorders. There exists, in particular, a great deal of interest in the correlation between blood serum levels and the severity of risk for cardiovascular disease. A sensitive, label-free, non-amplified and reusable electrochemical impedimetric biosensor for the detection of CRP in blood serum was developed herein based on controlled and coverage optimised antibody immobilization on standard polycrystalline gold electrodes. Charge transfer resistance changes were highly target specific, linear with log. CRP. concentration across a 0.5-50. nM range and associated with a limit of detection of 176. pM. Significantly, the detection limits are better than those of current CRP clinical methods and the assays are potentially cheap, relatively automated, reusable, multiplexed and highly portable. The generated interfaces were capable not only of comfortably quantifying CRP across a clinically relevant range of concentrations but also of doing this in whole blood serum with interfaces that were, subsequently, reusable. The importance of optimising receptor layer resistance in maximising assay sensitivity is also detailed. © 2012.

Determination of salicylate in blood serum using an amperometric biosensor based on salicylate hydroxylase immobilized in a polypyrrole-glutaraldehyde matrix

The use of an amperometric biosensor for the salicylate determination in blood serum is described. The biosensor is based on salicylate hydroxylase (EC 1.14.13.1) electropolymerized onto a glassy carbon-working electrode with polypyrrole and glutaraldehyde, to improve the biosensor lifetime. The hexacyanoferrate (II) was also incorporated to work as a redox mediator to minimize possible interferences. The salicylate is enzymatically converted to catechol, which is monitored amperometrically by its electrooxidation at +0.170 V versus SCE (saturated calomel electrode). Salicylate determination was carried out maintaining the ratio between β-NADH and salicylate at 4:1 (30°C). The amperometric response of the biosensor was linearly proportional to the salicylate concentration between 2.3 x 10-6 and 1.4 x 10-5 mol l- 1, in 0.1 mol l-1 phosphate buffer (pH 7.8), containing 0.1 mol l-1 KCl and 5.0 x 10-4 mol l-1 Na2H2EDTA, as supporting electrolyte. The recovery studies, in the presence of several interfering compounds, showed recoveries between 96.4 and 104.8%. The useful lifetime of the biosensor in the concentration range evaluated was at least 40 days, in continuous use. Blood serum samples analyzed by this biosensor showed a good correlation compared to the spectrophotometric method (Trinder) used as reference, presenting relative deviations lower than 7.0%. (C) 2000 Elsevier Science B.V.

Efeitos de probióticos sobre a digestibilidade, escore fecal e características hematológicas em cães

Avaliaram-se os efeitos da suplementação de dois tipos de probióticos para cães filhotes, que receberam dois tipos de dieta - de alta e de baixa qualidade -, sobre a digestibilidade dos nutrientes, escore fecal e parâmetros sanguíneos. Foram utilizados 18 animais, distribuídos em três tratamentos. No tratamento 1, controle, os cães receberam somente a ração; no tratamento 2, ração com probiótico 1 (Bifidobacterium) e, no tratamento 3, ração com probiótico 2 (Lactobacillus). O experimento foi dividido em duas fases. Verificaram-se que os valores médios do coeficiente de metabolizabilidade da energia bruta (CMEB) na fase 1, caracterizada pela troca da dieta Super Premium para a dieta Standard, apresentaram resultados significativos (P<0,05), sendo os melhores resultados obtidos nos animais que receberam o probiótico 2. Não houve diferenças significativas (P>0,05) para o escore fecal e para as análises hematológicas.

Renal excretion of zinc in normal individuals during zinc tolerance test and glucose tolerance test

The urinary excretion, renal clearance, and tubular reabsorption of zinc were investigated in 30 adult healthy subjects under basal conditions and during the zinc and glucose tolerance tests. After a 12h overnight fast, each subject was submitted to renal clearance of zinc. The procedures were performed between 8.00 and 12.00 a.m., after emptying the bladder and ingestion of 4 ml deionized water/kg body weight at 8.00 a.m. The first urine sample was collected at 10.00 a.m., and the second at 12.00 a.m. A dose of 110 mg ZnSO47H2O was administered orally to each subject, diluted in 20 ml deionized water, at time 0 min. Blood samples were collected from an antecubital vein at times -30, 0, and 30 min and at 30 min intervals up to 240 min. Glucose was administered intravenously (0.5 ml 50%/kg body weight) during the first 3 min of the test, and blood samples were collected from an unconstricted, contralateral, antecubital vein at times -30, 0, 3, 5, 10, 20, 30, 45, 60, and 90 min. The results showed that urinary zinc excretion, and renal zinc clearance were significantly higher during the zinc and glucose tolerance tests than in the control condition. On the other hand, renal zinc clearance was more elevated during tile glucose tolerance test than during the zinc tolerance test. Variations in zinc tubular reabsorption and glomerular filtration rate were not detected. The results suggest that urinary excretion and renal clearance of zinc in healthy subjects increase during acute zinc ingestion and glucose infusion, Although zinc ingestion raised urinary zinc excretion, glucose infusion was more effective in increasing renal zinc clearance. These normal parameters are important in the investigation of diabetic patients with serum and urine zinc changes.

Avaliação clínica e hematológica em bezerros Nelore infectados experimentalmente com isolados de Babesia bigemina das regiões Sudeste, Nordeste e Norte do Brasil

A comparative study was made regarding the clinical and hematological alterations caused by isolates of Babesia bigemina from southeastern, northeastern and northern Brazil in experimentally infected Nelore calves. Eighteen calves between 7 and 9 months of age, without antibodies against Babesia sp and raised free from ticks, were used. Three animals were previously inoculated with 2.0×109 parasitic erythrocytes (PE) for each stabilate. The other 15 calves were subdivided into three groups, with five animals each, that were subinoculated with 1.0×1010 PE of the respective isolates. The clinical and hematological alterations were evaluated by the determination of parasitaemia, haemogram, plasmatic fibrinogen, reticulocyte count, descriptive analysis of the bone marrow and erythrocytic osmotic fragility, for 30 days, totalizing seven moments of observation. The follow-up of the immunological response by the indirect fluorescent antibody test was carried out daily until the 10th day after inoculation (DAI) and after that, on the 15th, 20th, 25th and 30th DAI. A mild clinical manifestation of the disease was observed. The laboratory findings revealed low levels of parasitaemia; decrease of the erythrogram values; absence of reticulocytes, initial decrease in the total count of leukocytes, neutrophils and lymphocytes with a posterior elevation of the number of these cells; hypercellularity of the erythrocytic series and decrease of the myeloid: erythroid relation which was more accentuated between the 8th and 12th DAI, and an increase of the erythrocytic osmotic fragility in the groups inoculated with the Southeast and Northeast isolates. None of the three isolates of B. bigemina gave rise to the clinical characteristic form of the disease, although they induced an humoral immune response.

Características hematológicas, bioquímicas e biométricas de Piaractus mesopotamicus Holmberg, 1887 (Osteichthyes: Characidae) oriundos de cultivo intensivo

Hematological characteristics are important tools for diagnosis and prognostic features in fish population and they contribute to the comprehension of comparative physiology, phylogenetic relation, food habit and other ecological parameters. Red blood cell count (RBC), white blood cell count (WBC), thrombocytes counts, glucose, cortisol, hepatosomatic and splenosomatic relation were evaluated in Piaractus mesopotamicus. The values of hepatosomatic relation varied from 0.67% to 2.79% and splenosomatic relation from 0.011% to 0.108%. RBC 1.870 to 4.590 × 106/μL, hemoglobin concentration 5.7 to 12.1g/dL, haematocrit 24.0% to 40.0%, mean corpuscular volume 98.0 to 174.0 fL, mean corpuscular haemoglobin concentration 19.1 to 34.2g/dL, glucose 40.6 to 89.2mg/dL and cortisol 4.73 to 110.5ng/dL The number of total thrombocytes was 12,160.0 to 89,280.0/μL and WBC 2,019.0 to 47,473.0/μL. Differential count of leucocytes showed 54.1% limphocytes, 10.9% neutrophils, 10.3% monocytes, 9.9% PAS-positive granular leukocytes (PAS-GL), 6.7% eosinophils, 0.3% basophils and 7.8% immature leucocytes. Variation of these haematological characteristics in the same fish species was observed, being considered as the normal variation in the fish concerned. Hence it is incorrect to assume that biological characteristics in fishes are distributed normally.

Plication of the free wall of the left ventricle in dogs with doxorubicin-induced cardiomyopathy

Objective - To evaluate plication of the free wall of the left ventricle, which reduces the left ventricular area and volume, as a method to improve the left ventricular systolic function without cardiopulmonary bypass. Animals - 8 mixed-breed adult dogs. Procedure - Dilated cardiomyopathy (DCM) was induced in each dog by administration of doxorubicin (30 mg/m2, IV, q 21 d for 168 days). Two dogs died during induction of cardiomyopathy. Plication surgery was performed in 4 dogs. Two dogs did not ondergo to surgery (control group). Values for cardiac output (CO), 2-dimensional and M-mode echocardiography, arterial blood pressure, electrocardiography, blood cell counts, and serum biochemical analyses were recorded after induction of DCM (baseline) and 1, 2, 7, 15, 21, 30, 60, 90, 120, 150, and 180 days after plication surgery. Ambulatory ECG (Holter) recordings were conducted for 24 hours on the day of surgery. Results - 1 dog died after plication surgery. The remaining dogs undergoing ventricular plication had a significant improvement in CO, ejection fraction, and fractional shortening and reductions of left ventricular area and volume after surgery. Electrocardiographic and Holter recordings revealed premature ventricular complexes, which resolved without treatment during the first week after surgery. Clinical condition of the control dogs declined, and these 2 dogs died approximately 40 days after induction of cardiomyopathy. Conclusions and Clinical Relevance - Plication of the free wall of the left ventricle improved left ventricular systolic function in dogs with doxorubicin-induced cardiomyopathy. Additional studies are needed to evaluate its application in dogs with naturally developing DCM.

Monosodium glutamate in standard and high-fiber diets: Metabolic syndrome and oxidative stress in rats

Objective: This study determined the effects of adding monosodium glutamate (MSG) to a standard diet and a fiber-enriched diet on glucose metabolism, lipid profile, and oxidative stress in rats. Methods: Male Wistar rats (65 ± 5 g, n = 8) were fed a standard diet (control), a standard diet supplemented with 100 g of MSG per kilogram of rat body weight, a diet rich in fiber, or a diet rich in fiber supplemented with 100 g of MSG per kilogram of body weight. After 45 d of treatment, sera were analyzed for concentrations of insulin, leptin, glucose, triacylglycerol, lipid hydroperoxide, and total antioxidant substances. A homeostasis model assessment index was estimated to characterize insulin resistance. Results: Voluntary food intake was higher and feed efficiency was lower in animals fed the standard diet supplemented with MSG than in those fed the control, fiber-enriched, or fiber- and MSG-enriched diet. The MSG group had metabolic dysfunction characterized by increased levels of glucose, triacylglycerol, insulin, leptin, and homeostasis model assessment index. The adverse effects of MSG were related to an imbalance between the oxidant and antioxidant systems. The MSG group had increased levels of lipid hydroperoxide and decreased levels of total antioxidant substances. Levels of triacylglycerol and lipid hydroperoxide were decreased in rats fed the fiber-enriched and fiber- and MSG-enriched diets, whereas levels of total antioxidant substances were increased in these animals. Conclusions: MSG added to a standard diet increased food intake. Overfeeding induced metabolic disorders associated with oxidative stress in the absence of obesity. The fiber-enriched diet prevented changes in glucose, insulin, leptin, and triacylglycerol levels that were seen in the MSG group. Because the deleterious effects of MSG, i.e., induced overfeeding, were not seen in the animals fed the fiber-enriched diets, it can be concluded that fiber supplementation is beneficial by discouraging overfeeding and improving oxidative stress that is induced by an MSG diet. © 2005 Elsevier Inc. All rights reserved.

Importância da linha de base pré-exposição das colinesterases (ChEs) no controle ocupacional a praguicidas organofosforados e carbamatos

The most immediate toxic effects of a poisoning for organophosphate or carbamate are promoted by inhibition of cholinesterases (ChEs). However, a serious limitation to the diagnosis of mild organophosphate or carbamate poisoning and to preventive screening of organophosphate-exposed workers has been the large interindividual variability in ChEs activity. This makes it necessary to obtain a pre-exposure baseline measurement of enzyme activity as a basis for evaluating subsequent declines. Thus, the objective of this work is demonstrate the value of the pre-exposure baseline in the interpretation of the results of the ChEs activity of the carbamate or organophosphate-exposed workers. Using Nabb & Whitfield (1967) method to analyze blood of 48 workers the results have shown a erythrocyte cholinesterase range of 10.1 to 19.7 μmol/min/mL and plasm cholinesterase range of 2.2 to 6.9 μmol/min/mL. Despite the large interindividual variation presented, when one used the pre-exposure baseline it was possible to correlate symptoms of light poisoning of the exposure workers to carbamate with fall in the Er-ChE activity lesser then 30% of the pre-exposure baseline activity.

Abordagem diagnóstica dos pacientes com suspeita de trombose venosa profunda dos membros inferiores

Deep venous thrombosis is a relatively common disease, which can present pulmonary embolism as a complication in its acute phase, and later the post-thrombotic syndrome. Thus, diagnosis should be made as soon as possible, in order to prevent or minimize such complications. Several studies have shown that the symptoms and the clinical signs are inaccurate for the deep venous thrombosis diagnosis and that complementary exams are necessary. As an attempt to simplify the patients' assessment, Well et al., in 1997, developed a clinical prediction index that combines symptoms, signs and risk factors for deep venous thrombosis and managed to make a simpler approach through an association of this index with the complementary exams. Phlebography has been considered the gold standard of complementary exams. However, since it is an invasive exam and thus subject to complications, other diagnostic methods were introduced aiming at making the diagnostic approach simpler and less invasive. Doppler ultrasound, duplex scan, impedance plethysmography, computed tomography, and blood tests such as the D-dimer are some of the available methods for assessing the patient with suspicion of deep venous thrombosis. Among them, duplex scan has shown excellent accuracy and it is currently widely accepted as the first choice test for approaching the patient with deep venous thrombosis. Several authors have suggested an association of diagnostic methods to simplify and make the assessment of such patients more cost-effective, leading to the introduction of a wide range of diagnostic strategies. The different diagnostic methods used for assessing deep venous thrombosis are discussed, as well as a review of the literature on the accuracy, advantages and disadvantages of these methods. Copyright © 2005 by Sociedade Brasileira de Angiologia e Cirurgia Vascular.

Levedura íntegra e derivados do seu processamento em rações para tilápia do Nilo: Aspectos hematológicos e histológicos

The effects of inclusion of whole yeast, autolyzed yeast and yeast cell wall on hematological parameters and gut villus perimeter were evaluated in juvenile Nile tilapia, after 80 experimental days. Isoproteic (32.0% DP) and isoenergetic (3200 kcal DE kg -1) practical diets were supplemented with three levels of whole yeast or autolyzed yeast (1.0, 2.0 and 3.0%) and three levels of yeast cell wall (0.1, 0.2 and 0.3%), plus a control diet (with no test microingredients). Red blood cell count, hemoglobin concentration, total plasmatic protein, hematocrit percentage, mean corpuscular volume, mean corpuscular hemoglobin concentration and gut villus perimeter were evaluated. Variations on hematological parameters in animals fed diets with whole yeast; autolyzed yeast and yeast cell wall were observed to be within normal ranges for this species. There was significant influence (p<0.05) of different levels of yeast and derivatives on intestinal villus perimeter. Results showed that experimental period and proposed levels of whole yeast, autolyzed yeast and yeast cell wall do not provide undesirable alterations on standard hematological parameters to Nile tilapia and can be safely used to compound diets for this species. Results also showed that supplement of yeast cell wall provide higher intestinal villus perimeter.

Antisperm antibodies in infertile men and their correlation with seminal parameters

Aim: Antisperm antibodies (ASA) in males cause the autoimmune disease 'immune infertility'. The present study intended to detect the presence of ASA and their incidence in men with unexplained infertility, as well as to evaluate the correlation between the presence of ASA and semen parameter alterations. Methods: Blood and sperm assessment were collected to carry out a direct and indirect mixed antiglobulin reaction (MAR) test and semen analysis in infertile and fertile men from the University Hospital of the Faculty of Medicine, Sao Paulo State University, Sao Paulo. Results: In the MARtest, 18.18% of infertile men were positive for ASA. In fertile men, no positivity was found. A significant correlation between the presence of ASA with an increased white blood cell count plus a decreased hypoosmotic swelling test result was observed. Conclusions: The results indicate that ASA are involved in reduced fertility. It is not ASA detection per.se that provides conclusive information about the occurrence of damage to fertility. The correlation between infertility and altered seminal parameters reinforce the ASA participation in this pathology. © 2007 The Authors Journal compilation. © 2007 Japan Society for Reproductive Medicine.

Avaliação do estresse oxidativo no sangue de consumidores habituais de suco de laranja

Two biomarkers of oxidative stress were evaluated in healthy volunteers treated with daily doses of orange juice that is well known source of vitamin C and citric flavanones, which have been associated with antioxidant, anti-inflammatory and hypolipidemic effects. The antioxidant capacity in the blood serum was evaluated by studying the scavenging of 2,2'-diphenyl-1-picrylhydrazyl (DPPH) free radical, and also lipid peroxidation was evaluated by the thiobarbituric acid reacting substances (TBARS) assay. The results have shown that the regular consumption of orange juice increased 150% and 200% the serum antioxidant capacity for women and men respectively, but it was no significant change in the serum lipid peroxidation. In conclusion, the increase of flavonones and vitamin C in the body due to the regular intake of orange juice expressively improved the antioxidant capacity, but without significant effect on the lipid peroxidation.