Guanosine protects against behavioural and mitochondrial bioenergetic alterations after mild traumatic brain injury.

Traumatic brain injury (TBI) constitutes a heterogeneous cerebral insult induced by traumatic biomechanical forces. Mitochondria play a critical role in brain bioenergetics, and TBI induces several consequences related with oxidative stress and excitotoxicity clearly demonstrated in different experimental model involving TBI. Mitochondrial bioenergetics alterations can present several targets for therapeutics which could help reduce secondary brain lesions such as neuropsychiatric problems, including memory loss and motor impairment. Guanosine (GUO), an endogenous neuroprotective nucleoside, affords the long-term benefits of controlling brain neurodegeneration, mainly due to its capacity to activate the antioxidant defense system and maintenance of the redox system. However, little is known about the exact protective mechanism exerted by GUO on mitochondrial bioenergetics disruption induced by TBI. Thus, the aim of this study was to investigate the effects of GUO in brain cortical and hippocampal mitochondrial bioenergetics in the mild TBI model. Additionally, we aimed to assess whether mitochondrial damage induced by TBI may be related to behavioral alterations in rats. Our findings showed that 24 h post-TBI, GUO treatment promotes an adaptive response of mitochondrial respiratory chain increasing oxygen flux which it was able to protect against the uncoupling of oxidative phosphorylation (OXPHOS) induced by TBI, restored the respiratory electron transfer system (ETS) established with an uncoupler. Guanosine treatment also increased respiratory control ratio (RCR), an indicator of the state of mitochondrial coupling, which is related to the mitochondrial functionality. In addition, mitochondrial bioenergetics failure was closely related with locomotor, exploratory and memory impairments. The present study suggests GUO treatment post mild TBI could increase GDP endogenous levels and consequently increasing ATP levels promotes an increase of RCR increasing OXPHOS and in substantial improve mitochondrial respiration in different brain regions, which, in turn, could promote an improvement in behavioral parameters associated to the mild TBI. These findings may contribute to the development of future therapies with a target on failure energetic metabolism induced by TBI.

Can short-term exposure to copper and atrazine be cytotoxic to microalgae?

Aquatic environments can be easily contaminated due to anthropogenic activities that may affect local biota. Microalgae are abundant and have an important role on the food chain. Consequently, they stand out as promising models for studies of contaminants. This study investigated the cytotoxic effects of atrazine and copper (separate and mixture) exposure in microalgae Desmodesmus communis, as well as its cellular defense due to ABC (ATP-binding cassette) proteins activity against the xenobiotics. We analyzed two different ABC proteins activity pathways: P-gp, which is responsible for nonspecific substance efflux, and MRP that is associated with metals efflux. It was observed that the microalgae exposure to atrazine (90 nM) and copper (141 nM) has been considered cytotoxic. When contaminants were mixed, only the combination of both highest concentrations tested was cytotoxic. The P-gp blocker, verapamil, demonstrated that the contaminants tested caused proteins inhibition. However, the MK-571 (MRP blocker) did not block pump activity. There was an inverse relationship between ABC protein activity and cytotoxicity; non-cytotoxic conditions suggest increased activity of microalgae defense proteins.

In Vitro Adult Astrocytes are Derived From Mature Cells and Reproduce in Vivo Redox Profile.

Astrocytes are versatile cells involved in synaptic information processing, energy metabolism, redox homeostasis, inflammatory response, and structural support of the brain. Recently, we established a routine protocol of cultured astrocytes derived from adult and aged Wistar rats, which present several different responses compared to newborn astrocytes, commonly used to characterize the role of the astrocytes in the central nervous system. Previous studies hypothesized that astrocyte cultures prepared from adult animals derive from immature precursors present in the adult tissue throughout life. Since our group has already demonstrated that the glial functionality of adult astrocytes differs from newborn cultures, the aim of this study was to confirm that our in vitro astrocytes were derived from mature cells. Therefore, we evaluated cytoskeleton proteins, such as glial fibrillary acidic protein and vimentin, as well as Sox10, an essential marker of immature glial cells, in ex vivo tissue and in in vitro astrocytes from the same animals (1, 90, and 180 days old). In addition, we examined the mitochondrial functionality and the cellular redox homeostasis. Our results suggest that adult and aged astrocytes are derived from mature cells and that changes in mitochondrial parameters in ex vivo tissue were reproduced in in vitro astrocytes. J. Cell. Biochem. 118: 3111-3118, 2017. © 2017 Wiley Periodicals, Inc.

Olive oil as an alternative to boar semen cryopreservation / Aceite de oliva como una alternativa para la criopreservación de semen suino / O azeite de oliva como uma alternativa para a criopreservação do sêmen suíno

The aim of this study was to evaluate different concentrations of olive oil for cryopreservation of boar semen. A total of 21 eyaculates of 18 males with motility ≥70% were used. For freezing, the semen was diluted in treatments: control (Cont-egg yolk), "Koroneiki" oil in egg yolk at 0.25% (A025), 0.50% (A050), 0.75% (A075) and 1.0% (A10). Straws with 5.107 sperm/ml were frozen and stored in liquid nitrogen. Thawing was done at 37ºC for 30 seconds and motility, mitochondrial functionality, DNA integrity, oocite penetration rate and the number of sperm per oocite (in the in vitro penetration trial) were evaluated. The variables were compared using the Kruskal-Wallis test. Even though no statistical difference was found between control and treatments containingolive oil (p>0.05), the treatment with 0.25% oil concentration showed tendences towards sperm protection, outperforming the controls in mitochondrial functionality and preservation of the fertilizing capabilities in the in vitro penetration trial. This is why, olive oil could represent an alternative to help cryopreservation of boar semen.

El objetivo de este estudio fue evaluar diferentes concentraciones de aceite de oliva para la criopreservación de semen de verraco. Se utilizaron 21 eyaculados de 18 machos con motilidad igual o superior al 70%. Para la congelación, el semen se diluyó en los tratamientos: control (Cont) (lactosa yema), 0,25% (A025); 0,50% (A050); 0,75% (A075) y 1,0% (A10) de aceite de la variedad "Koroneiki" en lactosa yema. Pajas con 5.107 espermatozoides / ml, fueron congeladas y almacenadas en nitrógeno líquido. La descongelación se produjo a 37°C durante 30 segundos y se evaluó: la motilidad, la funcionalidad de la mitocondria, la integridad del ADN, la tasa de penetración de ovocitos y el número de espermatozoides por ovocito (en el ensayo de penetración in vitro). Las variables se compararon mediante la prueba de Kruskal-Wallis. Aunque no hubo diferencia estadística entre el control y los tratamientos que contienen aceite de oliva (p> 0,05), la concentración de 0,25% de este aceite mostró tendencias de protección de esperma, superando el control en la funcionalidad de las mitocondrias y en la preservación de la capacidad fertilizante en el ensayo de penetración in vitro. Por lo tanto, el aceite de oliva puede representar una alternativa para ayudar en la criopreservación de semen porcino

O objetivo de este estudo foi avaliar diferentes concentrações de azeite de oliva para a criopreservação de sêmen suíno. Utilizaram-se 21 ejaculados de 18 machos com motilidade igual ou superior ao 70%. Para o congelamento, o sêmen se dilui-o nos tratamentos: controle (Cont) (lactose-gema), 0,25% (A025); 0,50% (A050); 0,75% (A075) e 1,0% (A10) de azeite da variedade "Koroneiki" em lactose-gema. Palhetas com uma dose de 5.107 espermatozoides/ ml foram congeladas e armazenadas em nitrogênio liquido. O descongelamento se produz a 37°C durante 30 segundos e se avaliaram: motilidade, funcionalidade da mitocôndria, integridade do DNA, taxa de penetração de ovócitos e o número de espermatozoides por ovócito (no teste de penetração in vitro). As variáveis se compararam mediante o teste de Kruskal-Wallis. Ainda que não houve diferença estatística entre o controle e os tratamentos que continham azeite de oliva (p> 0,05), a concentração de 0,25% de azeite de oliva teve tendências de proteção do esperma, superando o controle na funcionalidade das mitocôndrias e na preservação da capacidade fertilizante no teste de penetração in vitro. Assim sendo, o azeite de oliva pode representar uma alternativa para ajudar na criopreservação do sêmen suíno.

Functional integrity of Colossoma macropomum (Cuvier, 1816) sperm cryopreserved with enriched extender solutions

Cryoprotectant solutions are used to protect the sperm from alterations caused by the low temperature in the cryopreservation process. We evaluated the quality of Colossoma macropomum semen after freezing, using dimethyl sulfoxide (DMSO) as a cryoprotectant, combined with two extender solutions (T1 - Solution 1: Glucose 90.0 g/L, Sodium Citrate 6.0 g/L, EDTA 1.5 g/L, Sodium Bicarbonate 1.5 g/L, Potassium Chloride 0.8 g/L, Gentamycin Sulphate 0.2 g/L, and T2 - Solution 2: Glucose 90.0 g/L, ACP(r)-104 10.0 g/L). Motility rate and motility time did not differ between T1 and T2 and were lower than fresh semen. The number of normal sperm was significantly different in treatments T1 (15.1%) and T2 (21.9%), and both showed a reduction in the percentage of normal sperm compared to fresh semen (57.4%). The values found for the rates of fertilization and hatching, mitochondrial functionality and sperm DNA, did not differ between the treatments (T1 and T2). Regarding membrane integrity, there was a higher percentage of spermatozoa with intact membranes in T1 (53.4%) than T2 (43.7%). The extender solutions, combined with 10% DMSO, maintained the sperm DNA intact in almost all the C. macropomumsperm cells, however there was a loss in their functionality.(AU)

As soluções crioprotetoras são utilizadas para proteger os espermatozoides das alterações causadas por baixas temperaturas durante o processo de criopreservação. Avaliamos a qualidade do sêmen de Colossoma macropomumapós o congelamento, utilizando dimetilsulfóxido (DMSO) como crioprotetor, combinado com duas soluções diluidoras (T1 - Solução 1: Glicose 90,0 g/L, Citrato de Sódio 6,0 g/L, EDTA 1,5 g/L, Bicarbonato de Sódio 1,5 g/L, Cloreto de Potássio 0,8 g/L, Sulfato de Gentamicina 0,2 g/L, e T2 - Solução 2: Glicose 90,0 g/L, ACP(r)-104 10,0 g/L). A taxa de motilidade (%) e o tempo de motilidade (s) não diferiram entre T1 e T2, porém foram mais baixos do que no sêmen fresco. O número de espermatozoides normais foi significativamente diferente nos tratamentos T1 (15,1%) e T2 (21,9%), e ambos mostraram uma redução na porcentagem de espermatozoides normais, comparado ao sêmen fresco (57,4%). Os valores encontrados para as taxas de fertilização e eclosão, funcionalidade mitocondrial e DNA do esperma, não diferiram entre os tratamentos (T1 e T2). Para a integridade da membrana, houve uma porcentagem mais elevada de espermatozóides com a membrana intacta em T1 (53,4%) do que T2 (43,7%). As soluções diluentes combinadas com DMSO a 10% preservaram o DNA espermático intacto em quase todas as células do sêmen de C. macropomum, mas houve perda na funcionalidade dos mesmos.(AU)

Functional integrity of Colossoma macropomum (Cuvier, 1816) sperm cryopreserved with enriched extender solutions

Cryoprotectant solutions are used to protect the sperm from alterations caused by the low temperature in the cryopreservation process. We evaluated the quality of Colossoma macropomum semen after freezing, using dimethyl sulfoxide (DMSO) as a cryoprotectant, combined with two extender solutions (T1 - Solution 1: Glucose 90.0 g/L, Sodium Citrate 6.0 g/L, EDTA 1.5 g/L, Sodium Bicarbonate 1.5 g/L, Potassium Chloride 0.8 g/L, Gentamycin Sulphate 0.2 g/L, and T2 - Solution 2: Glucose 90.0 g/L, ACP(r)-104 10.0 g/L). Motility rate and motility time did not differ between T1 and T2 and were lower than fresh semen. The number of normal sperm was significantly different in treatments T1 (15.1%) and T2 (21.9%), and both showed a reduction in the percentage of normal sperm compared to fresh semen (57.4%). The values found for the rates of fertilization and hatching, mitochondrial functionality and sperm DNA, did not differ between the treatments (T1 and T2). Regarding membrane integrity, there was a higher percentage of spermatozoa with intact membranes in T1 (53.4%) than T2 (43.7%). The extender solutions, combined with 10% DMSO, maintained the sperm DNA intact in almost all the C. macropomumsperm cells, however there was a loss in their functionality.

As soluções crioprotetoras são utilizadas para proteger os espermatozoides das alterações causadas por baixas temperaturas durante o processo de criopreservação. Avaliamos a qualidade do sêmen de Colossoma macropomumapós o congelamento, utilizando dimetilsulfóxido (DMSO) como crioprotetor, combinado com duas soluções diluidoras (T1 - Solução 1: Glicose 90,0 g/L, Citrato de Sódio 6,0 g/L, EDTA 1,5 g/L, Bicarbonato de Sódio 1,5 g/L, Cloreto de Potássio 0,8 g/L, Sulfato de Gentamicina 0,2 g/L, e T2 - Solução 2: Glicose 90,0 g/L, ACP(r)-104 10,0 g/L). A taxa de motilidade (%) e o tempo de motilidade (s) não diferiram entre T1 e T2, porém foram mais baixos do que no sêmen fresco. O número de espermatozoides normais foi significativamente diferente nos tratamentos T1 (15,1%) e T2 (21,9%), e ambos mostraram uma redução na porcentagem de espermatozoides normais, comparado ao sêmen fresco (57,4%). Os valores encontrados para as taxas de fertilização e eclosão, funcionalidade mitocondrial e DNA do esperma, não diferiram entre os tratamentos (T1 e T2). Para a integridade da membrana, houve uma porcentagem mais elevada de espermatozóides com a membrana intacta em T1 (53,4%) do que T2 (43,7%). As soluções diluentes combinadas com DMSO a 10% preservaram o DNA espermático intacto em quase todas as células do sêmen de C. macropomum, mas houve perda na funcionalidade dos mesmos.