The aim of this investigation was to evaluate the effects of 3 overtraining (OT) protocols on the glial activation and apoptosis in the spinal cords of mice. Rodents were divided into control (C; sedentary mice), overtrained by downhill running (OTR/down), overtrained by uphill running (OTR/up) and overtrained by running without inclination (OTR). The incremental load test, ambulation test, exhaustive test and functional behavioural assessment were used as performance evaluation parameters. 36 h after the exhaustive test, the dorsal and ventral parts of the lumbar spinal cord (L4-L6) were dissected for subsequent protein analysis by immunoblotting. The OT protocols led to similar responses of some performance parameters. The ventral glial fibrillary acidic protein (GFAP) protein levels were diminished in the OTR/up and OTR compared to CT and OTR/down groups. The ventral ionized calcium binding adaptor molecule 1 (Iba-1), and the dorsal GFAP and Iba-1 protein levels were increased in the OTR/down compared to the other groups. The ratio between the cleaved capase-3/caspase-3 and cleaved caspase-9/caspase-9 measured in the spinal cord were not sensitive to the OT protocols. In summary, the OTR/down activated the glial cells in the motor (i. e. Iba-1) and sensory (i. e. GFAP and Iba-1) neurons without leading to apoptosis.
This study evaluated the activity of lactoferrin on the multiplication of probiotic Lactobacillus casei in vitro and in Minas fresh cheese. Growth curves of L. casei in BHI broth containing 1 mg/mL and 2 mg/mL of lactoferrin were performed. Additionally, Minas fresh cheeses added of L. casei (control) and added of L. casei and lactoferrin (2 mg/g and 4 mg/g) were produced and stored at 5°C during 28 days. Cheeses were analyzed for pH, titratable acidity and enumeration of L. casei and psychotropic microorganisms in days 1, 7, 14, 21 and 28 and for the centesimal composition in day 1. The experiment was repeated three times, and data were analyzed by ANOVA and Tukey test at 5% significance level. When tested in vitro, L. casei multiplication was stimulated by lactoferrin at a concentration of 2 mg/ mL, but this activity has not been verified in the cheese, even in that added by lactoferrin at 4 mg/g. Psychotropic population in the cheeses added by lactoferrin did not differ from control cheese (P>0.05), demonstrating that no antimicrobial activity occurred in the products. More studies should be performed to evaluate the antimicrobial effects of lactoferrin in foods, since they have more variables that may affect the activity of this protein when comparing to in vitro tests. Key words: Antimicrobial activity, biologic activity, cheese, milk products, probiotics ResumoO presente estudo avaliou a atividade da lactoferrina sobre a multiplicação de Lactobacillus casei probiótico in vitro e no queijo Minas frescal. As curvas de crescimento de L. casei em caldo BHI contendo lactoferrina nas concentrações de 1 mg/mL and 2 mg/mL foram construídas. Adicionalmente, queijos Minas frescal contendo somente L. casei (controle) e L. casei e a lactoferrina (2 mg/g e 4 mg/g) foram produzidos e armazenados a 5°C durante 28 dias. Foram realizadas as análises de pH, acidez livre titulável e enumeração de L. casei e microrganismos psicrotróficos no dia 1 (dia da produção) e após 7, 14, 21 e 28 dias de armazenamento. Além disso, no dia 1, foram realizadas as análises de composição centesimal. Os experimentos foram realizados em triplicata e os dados obtidos foram analisados utilizando-se ANOVA e teste de Tukey a 5% de significância. Nos ensaios in vitro a multiplicação de L. casei foi estimulada pela presença da lactoferrina na concentração de 2 mg/mL. Entretanto, esta atividade não foi observada no queijo, mesmo quando a concentração de 4 mg/g de lactoferrina foi utilizada. A população de microrganismos psicrotróficos presentes nos queijos adicionados de lactoferrina não diferiram das populações observadas para os queijos controle (P>0,05), demostrando que não houve atividade antimicrobiana nos produtos. Estudos futuros devem ser conduzidos para avaliar os efeitos antimicrobianos da lactoferrina em alimentos, pois são muitas as variáveis que afetam a atividade desta proteína quando comparados aos resultados obtidos in vitro.
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