Heteroplasmy in Hair: Study of Mitochondrial DNA Third Hypervariable Region in Hair and Blood Samples

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The placenta as a barrier for toxic and essential elements in paired maternal and cord blood samples of South African delivering women

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Evaluation of a portable clinical analyzer for the determination of blood gas partial pressures, electrolyte concentrations, and hematocrit in venous blood samples collected from cattle, horses, and sheep

Objective To compare results reported for blood gas partial pressures, electrolyte concentrations, and Hct in venous blood samples collected from cattle, horses, and sheep and analyzed by use of a portable clinical analyzer (PCA) and reference analyzer (RA).Animals-Clinically normal animals (24 cattle, 22 horses, and 22 sheep).Procedures-pH; Pco(2); Po(2); total carbon dioxide concentration; oxygen saturation; base excess; concentrations of HCO(3)(-), Na(+), K(+), and ionized calcium; Hct; and hemoglobin concentration were determined with a PCA. Results were compared with those obtained for the same blood sample with an RA. Bias (mean difference) and variability (95% confidence interval) were determined for all data reported. Data were also subjected to analyses by Deming regression and Pearson correlation.Results-Analysis of Bland-Altman plots revealed good agreement between results obtained with the PCA and those obtained with the RA for pH and total carbon dioxide concentration in cattle, K(+) concentration in horses and sheep, and base excess in horses. Except for Na(+) concentration and Hct in horses and sheep, correlation was good or excellent for most variables reported.Conclusions and Clinical Relevance-Data from blood gas and electrolyte analyses obtained by use of the PCA can be used to evaluate the health status of cattle, horses, and sheep. Furthermore, the handheld PCA device may have a great advantage over the RA device as a result of the ability to analyze blood samples on farms that may be located far from urban centers. (Am J Vet Res 2010;71:515-521)

Direct and Simultaneous Determination of Cd Cu, and Se in Blood Samples by Graphite Furnace Atomic Absorption Spectrometry

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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.

Evaluation of zebu nellore cattle blood samples using the Cell-Dyn 3500 hematology analyzer

The Cell-dyn 3500 is a multiparameter flow cytometer, which may analyze samples from several species performing several simultaneous analyses. It is able to perform white blood cells, red blood cells and platelet counts, besides differential leukocyte counts, packed cell volume and hemoglobin determination. Cell-Dyn 3500 performs total leukocyte count both optically and by impedance. The equipment may choose one or other method, based on the reliability of the results. Erythrocyte and platelet counts are determined by impedance. Leukocyte differentiation is based on an optical principle, using separation in multiangular polarized light. The objective of this study was to compare the results of complete blood count of Zebu Nellore heifers from Cell-dyn 3500, with those obtained from a semi-automated cell counter (Celm CC 510) and the manual technique. Blood samples were collected from the jugular vein in 5 mL EDTA vacuum tubes from 58 Nellore heifers, at 24 months of age. Samples were processed in parallel in the three different techniques. Results were analyzed using paired t test, Pearson's correlation and the Bland-Altmann method. There was a strong correlation for all parameters analyzed by Cell-Dyn 3500, manual method and semi-automated cell counter, except for basophils and monocytes counts. These results confirm that this analyzer is reliable for blood samples analysis of zebu cattle.

Improving DNA extraction quality by the salting-out method to prepare blood samples for real-time PCR.

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Alcohol levels in cerebrospinal fluid and blood samples from patients under pathological conditions

We measured alcohol levels by the Cordebard method in 148 CSF samples from individuals who had abstained from alcohol for at least 7 days prior to the beginning of the study. Each blood sample was accompanied by a CSF sample from the same patient. CSF samples found to be normal after analysis were used as controls. Mean alcohol concentration in blood did not differ significantly between the control group and the groups with altered CSF. The group with altered CSF had statistically higher alcohol levels in CSF than in blood. CSF lactate, glucose and protein levels were not correlated with alcohol level. The results suggest the presence of endogenous alcohol in the CSF, with levels increasing in the presence of pathological processes involving the nervous system.

Ehrlichia canis morulae and DNA detection in whole blood and spleen aspiration samples

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Levels of DNA damage in blood leukocyte samples from non-diabetic and diabetic female rats and their fetuses exposed to air or cigarette smoke

dThe objective of the present study was to evaluate DNA damage level in blood leukocytes from diabetic and non-diabetic female Wistar rats exposed to air or to cigarette smoke, and to correlate the findings with levels of DNA damage detected in blood leukocyte samples from their fetuses. A total of 20 rats were distributed into four experimental groups: non-diabetic (control; G1) and diabetic exposed to filtered air (G2): non-diabetic (G3) and diabetic (G4) exposed to cigarette smoke. Rats placed into whole-body exposure chambers were exposed for 30 min to filtered air (control) or to tobacco smoke generated from 10 cigarettes, twice a day, for 2 months. Diabetes was induced by a pancreatic beta-cytotoxic agent, streptozotocin (40 mg/kg b.w.). At day 21 of pregnancy, each rat was anesthetized and humanely killed to obtain maternal and fetal blood samples for genotoxicity analysis using the alkaline comet assay. G2, G3 and G4 dams presented higher DNA damage values in tail moment and tail length as compared to G1 group. There was a significant positive correlation between DNA damage levels in blood leukocyte samples from G2 and G3 groups (tail moment); G3 and G4 groups (tail length) and G3 group (tail intensity) and their fetuses. Thus, this study showed the association of severe diabetes and tobacco cigarette smoke exposure did not exacerbate levels of maternal and fetal DNA damages related with only diabetes or cigarette smoke exposure. Based on the results obtained and taking into account other published data, maternal diabetes requires rigid clinical control and public health and education campaigns should be increased to encourage individuals, especially pregnant women, to stop smoking. (C) 2008 Elsevier B.V. All rights reserved.

Long-term facial artery catheter implantation for serial arterial blood sampling and invasive arterial blood pressure measurement in horses

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The effect of offering an energy and protein supplement to grazing canchim beef cows either postpartum or both pre- and postpartum on lipid blood metabolites and folliculogenesis

This study was designed to evaluate the effect of nutritional supplementation offered during the pre- and postpartum periods on serum cholesterol, triglycerides and total lipids of Canchim beef cows and their relationship with folliculogenesis. Thirty cows with predicted calving date between September and October, kept in pastures of Brachiaria brizantha cv. Marandu together with their calves, were randomly distributed into three experimental groups: the first received only a mineral mixture (Control Group, CG); the second group received a concentrate with 16% crude protein/kg dry matter (DM) and 3000 kcal digestible energy/kg DM offered for 45 days prepartum and 120 days postpartum (PREG); the third group received the concentrate from parturition until the 120th day postpartum (POSG). Consumption was estimated at 1% of body weight, and each cow received approximately 4.0 kg/day (fresh weight) of supplement. Blood samples were taken and an ultrasound examination of the ovaries was performed twice a week until the 60th day postpartum. The body condition score (BCS) and the weight of the cows were recorded at 15-day intervals from calving until the 60th day postpartum. Data are presented as mean +/- SEM. Mean weight and BCS at calving were, respectively, 448 +/- 54.9 kg and 6.2 +/- 0.25 (PREG); 432 +/- 71.1 kg and 5.5 +/- 0.69 (POSG); and 434 +/- 66.4 kg and 5.5 +/- 0.69 (CG). Total cholesterol (TC), triglycerides (TRIG) and total lipids (TLIP) were measured using colorimetry until the 60th day postpartum. TC averages were PREG 186 +/- 62.6 mg/dL, POSG 159 +/- 43.1 mg/dL and CG 133 +/- 35.1 mg/dL (P < 0.05). For TRIG, the means were PREG 29 +/- 11.3 mg/dL (P < 0.05), POSG 24 +/- 8.1 mg/dL and CG 26 +/- 12.1 mg/dL (P > 0.05). Serum concentrations of TLIP were PREG 588 +/- 145.6 mg/dL, POSG 512 +/- 137.6 mg/dL and CG 452 +/- 122.4 mg/dL (P < 0.05). The first dominant follicle (DF) was identified on Day 21 +/- 10.3 (PREG), 36 +/- 28.5 (POSG) and 51 +/- 32.8 (CG) after calving. The difference between PREG and CG was significant (P < 0.05). TC was positively correlated with the calving to first estrus interval (P < 0.05). Results showed that nutritional supplementation before parturition assured good body condition at calving and suggested that it was effective at increasing cholesterol availability to maintain ovarian follicle function and to favor earlier resumption of ovarian activity. (C) 2010 Published by Elsevier B.V.

Effect of oils sources on blood lipid parameters of commercial laying hens

The experiment was carried out to verify if total cholesterol, HDL-cholesterol and triacylglicerol plasma levels are affected when laying hens are fed rations containing different dietary oil sources. One hundred sixty 50 week-old hens, assigned to four treatments with five replicates using 8 hens per replicate were used. The experimental period was of 84 days divided in 3 cycles of 28 days each. In the last day of each cycle, blood samples of 2 hens per replicate were randomly choose and blood samples were collected. on the other hand, blood was also collected at 7 am, 11 am and 3 pm aiming to study the daily changes of these lipids. Blood lipid parameters were not affected by different dietary oil sources (p > 0.05); however, HDL-cholesterol did change during the day, giving evidence that this lipid is indeed involved in the egg yolk formation.

Intraocular pressure and its correlation with blood gas parameters in crested caracaras (Caracara plancus) anesthetized with isoflurane and sevoflurane

Avaliou-se a pressão intra-ocular (PIO) e estimaram-se as correlações entre PIO e pressão de dióxido de carbono (PaCO2) e pH arterial de cinco caracarás (Caracara plancus), anestesiados com isofluorano (ISO) ou sevofluorano (SEV). Valores basais da PIO foram aferidos em ambos os olhos (M0). Cateterizou-se previamente a artéria braquial para obtenção de parâmetros hemogasométricos e cardiorrespiratórios. Anestesia foi induzida com ISO a 5V% e mantida por 40 minutos com 2,5V%. PIO e amostras de sangue foram avaliadas em diferentes momentos até o final do procedimento. Após recuperação, uma segunda anestesia foi realizada com SEV a 6% e mantida com 3,5%. Os parâmetros foram aferidos nos mesmos momentos estabelecidos previamente. A PIO decresceu significativamente (P=0,012) de M0 em todos os momentos e não houve diferença estatística entre ISO e SEV. Correlações significativas entre PIO e PaCO2 e entre PIO e pH sangüíneo foram observadas apenas para a anestesia com SEV. O pH sangüíneo decresceu paralelamente a PIO, enquanto a PaCO2 aumentou, em carcarás anestesiados com isofluorano e sevofluorano.

Organic and inorganic sources of zinc, copper and selenium in diets for dairy cows: intake, blood metabolic profile, milk yield and composition

The present study was carried out with the objective of evaluating the effects of feeding dairy cows with organic or inorganic sources of zinc (Zn), copper (Cu) and selenium (Se) on blood concentrations of these minerals, blood metabolic profiles, nutrient intake and milk yield and composition. Nineteen Holstein cows were selected and randomly assigned to two groups for receiving organic (n = 9) or inorganic (n = 10) sources of Zn, Cu and Se from 60 days before the expected date of calving to 80 days of lactation. Samples of feed, orts and milk were collected for analysis. Body condition score (BCS) was determined and blood samples were collected for analysis of Zn, Cu and Se concentrations, as well as for metabolic profile. Supplying organic or inorganic sources of Zn, Cu, and Se did not affect dry matter and nutrient intake, blood metabolic profile, milk yield and composition, plasma concentration of these minerals, and BCS or change the BCS in cows from 60 days before the expected date of calving to 80 days of lactation. An effect of time was observed on all feed intake variables, plasma concentrations of Zn and Se, milk yield, milk protein content, BCS and change in BCS.