Neutralized and stained nuclei (from random 100-cells fields) wer

Neutralized and stained nuclei (from random 100-cells fields) were blindly analyzed by fluorescence microscopy (200×). Cells were scored

from 0 (undamaged) to 4 (maximally damaged), according to the tail intensity (size and shape), resulting in a single DNA damage score Afatinib in vivo for each cell, and consequently, for each group. Thus, every group could be ranged for damage index with a value from 0 (all cells no tail, 100 cells × 0) to 400 (all cells with maximally long tails, 100 cells × 4) (Collins et al., 1997). The index of DNA damage was calculated by multiplying the number of cells by its own index score and then summed up its results. Total protein content was determined by the modified method of Lowry as previously described (Peterson, 1977), using BSA as standard. Data are reported as mean ± standard

error mean (S.E.M.) and were analyzed by Student’s t-test. Values of P < 0.05 were considered significant. All analyses were performed using the SPSS program, Version 12.0 (SPSS, Chicago, IL). This work was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), FINEP/ Rede IBN01.06.0842-00 and National Institute of Science and Technology for Excitotoxicity and Neuroprotection (INCT-EN). "
“Stroke is the third leading cause of death in industrialized countries (Lewis et al., 2008) and the most Belnacasan order frequent cause of permanent disability in adults worldwide (Donnan et al., 2008). Three months following a stroke, 15–30% of stroke survivors are permanently disabled and 20% require institutional care. Deficits can include partial paralysis, difficulties with memory, thinking, language, and movements. STK38 In the western world, over 70% of individuals experiencing a stroke are over 65 years of age. Since life expectancy continues to

grow, the absolute number of individuals with stroke will further increase in the future (Lakhan et al., 2009). Transient global ischemia arises as a consequence of cardiac arrest and causes selective, delayed death of hippocampal CA1 neurons in humans and can produce serious neurobiological sequellae of which cognitive deficits are most prominent (Lo et al., 2003, Moskowitz et al., 2010, Tanaka et al., 2000 and Merchenthaler et al., 2003; Etgen et al., 2010). Over the last decade, data from many studies support the idea that estrogens provide neuroprotective effects in a variety of focal and global ischemia models (Lebesgue et al., 2009, Merchenthaler et al., 2003, Garcia-Segura et al., 2001, Toran-Allerand, 2004 and Shughrue and Merchenthaler, 2003). The potent feminizing hormone, 17 beta-estradiol (E2), is neuroprotective in a host of cell and animal models of stroke and neurodegenerative diseases.

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