ABSTRACT
The study included the purification of glutathione peroxidase enzyme (GPX) in the serum of women with breast cancer, which involved 60 samples of serum from women with breast cancer, and 30 samples from healthy individuals. The results of the study showed a significant decrease at a probability level of p<0.0001 for the activity of the GPX enzyme in the serum of women with breast cancer. Additionally, the GPX enzyme was purified from the serum of women with breast cancer through precipitation with ammonium sulfate and dialysis, and the use of DEAE-Cellulose ion exchange chromatography and gel filtration chromatography using Sephadex G-100, where a main protein band was separated, which was relied upon in determining the optimal conditions for the partially purified enzyme. The optimal conditions for the partially purified enzyme from the serum of women with breast cancer were determined and the highest activity was for the substrate concentration of 0.1 mM H2O2. The maximum speed Vmax was 3.125IU/L and the Michaelis-Menten constant Km was 0.0179 M using Lineweaver-Burk plot, the optimal pH was at 8.5, temperature at 37°C, and the highest activity time was at 5 minutes.
Subject(s)
Breast Neoplasms , Glutathione Peroxidase , Humans , Female , Breast Neoplasms/enzymology , Glutathione Peroxidase/blood , Glutathione Peroxidase/isolation & purification , Glutathione Peroxidase/chemistry , Hydrogen-Ion Concentration , Hydrogen Peroxide/chemistry , Middle Aged , Kinetics , Temperature , Chromatography, Ion Exchange , Chromatography, Gel , AdultABSTRACT
Increased oxidative stress during prolonged endurance exercises may result in muscle damage, fatigue and decreased performance. An adequate stress response during training is critical to obtain improved results and high animal welfare standards. The aim of this study was to evaluate the red blood cell haemolysate concentrations of superoxide dismutase (SOD), glutathione peroxidase (GPx), reduced glutathione (GSH) and catalase (CAT) and the plasma concentrations of malondialdehyde (MDA) from endurance horses in different distances at high speed in a tropical climate. Fifteen horses were tested; five at 160km (18.54 - 17.16km/h race speed), five at 120km (21.53 - 17km/h race speed) and five at 80km (20.06 - 18.01km/h race speed). Blood samples were collected at rest, immediately after and three hours after the horses left the final vet check and three, seven and fourteen days after the race. No significant increases (P > 0.05) in the levels of SOD, GPx, GSH, CAT or MDA were observed for any of the times or distances examined. Based on these observations, we conclude that reactive oxygen species (ROS) formation during exercise evokes specific adaptations, such as increased antioxidant/oxidative damage-repairing enzyme activity, increased resistance to oxidative stress and lower levels of oxidative damage...
Aumento do estresse oxidativo durante o exercício prolongado pode resultar em fadiga muscular, lesões e diminuição do desempenho. Uma adequada resposta a esse estresse durante o treinamento é fundamental para a obtenção de melhores resultados e bem-estar dos animais. O objetivo deste estudo foi avaliar a concentração de superóxido dismutase (SOD), glutationa peroxidase (GPx), glutationa reduzida (GSH) e catalase (CAT) no hemolisado sanguíneo e malondialdeído (MDA) plasmático em cavalos de enduro correndo em diferentes distâncias, com alta média de velocidade, em clima tropical. Quinze cavalos foram testados, cinco em 160km (18.54-17.16km/h), cinco em 120km (21.53-17km/h) e cinco em 80km (20.06-18.01km/h). Amostras de sangue foram coletadas em repouso, imediatamente e três horas depois que os cavalos passaram pela inspeção veterinária final e três, sete e 14 dias após a corrida. Não houve aumentos significativos (P>0,05) dos níveis de SOD, GPx, GSH, CAT ou MDA em nenhum tempo nem distâncias analisadas. Com base nessas observações, pode-se concluir que as espécies reativas de oxigênio (ROS) formadas durante o exercício provocam adaptações específicas, tais como atividade antioxidante aumentada da enzima, maior resistência ao estresse oxidativo e menores níveis de danos oxidativos...
Subject(s)
Animals , Catalase/isolation & purification , Horses/metabolism , Oxidative Stress/physiology , Glutathione Peroxidase/isolation & purification , Glutathione/isolation & purification , Malondialdehyde/isolation & purification , Superoxide Dismutase/isolation & purification , Physical Conditioning, Animal/adverse effects , Running/physiology , Physical Exertion , Veterinary Sports MedicineABSTRACT
To study the deficiency of minerals and its relationship with Paratuberculosis, blood, serum, and fecal samples were obtained from 75 adult bovines without clinical symptoms of the disease and from two bovines with clinical symptoms of the disease, from two beef herds with a previous history of Paratuberculosis in the Province of Buenos Aires, Argentina. Serum samples were processed by ELISA and feces were cultured in Herrolds medium. Copper, zinc and iron in serum were quantified by spectrophotometry and selenium was measured by the activity of glutathione peroxidase. We also determined copper, zinc, iron and molybdenum concentrations in pastures and the concentration of sulfate in water. Mycobacterium avium subsp paratuberculosis (Map) was isolated from 17.3% of fecal samples of asymptomatic animals and from the fecal samples from the two animals with clinical symptoms. All the Map-positive animals were also ELISA-positive or suspect, and among them, 84.6% presented low or marginal values of selenium and 69.2% presented low or marginal values of copper. The two animals with clinical symptoms, and isolation of Map from feces and organs were selenium-deficient and had the lowest activity of glutathione peroxidase of all the animals from both herds. All the animals negative to Map in feces and negative to ELISA had normal values of Se, while 13.8% of animals with positive ELISA or suspect and culture negative presented low levels of Se. Half of the animals that were negative both for ELISA and culture in feces were deficient in copper but none of them presented low values of selenium. The content of molybdenum and iron in pasture was high, 2.5 ppm and 1.13 ppm in one herd and 2.5 ppm and 2.02 ppm in the other, respectively, whereas the copper:molybdenum ratio was 1.5 and 5.2, respectively. These results do not confirm an interaction between imbalances of the micronutrients and clinical Paratuberculosis, but show evidence of the relationship between selenium...