Resistance Training and Ovariectomy: Antagonic Effects in Mitochondrial Biogenesis Markers in Rat Skeletal Muscle.

Estrogen reduction is associated with a decline in skeletal muscle mitochondrial biogenesis. Molecular events associated with improvements in markers of mitochondrial biogenesis after resistance training and estradiol replacement are unknown. This study aimed to investigate the effects of ovariectomy, resistance training, and estradiol replacement on markers of mitochondrial biogenesis and protein expression related to oxidative capacity in the rat gastrocnemius pool. Estradiol replacement was performed using Silastic(®) capsules. During the 12-week resistance training, animals climbed a ladder with weights attached to their tails. Gene expression was analysed by RT-PCR, and protein content was determined by western blotting. Ovariectomy decreased the gene expression of the mitochondrial biogenesis markers PGC-1α (~73%), NRF-1 (~44%), and TFAM (~53%) (p<0.05) and decreased the protein expression of phosphorylated AMPK, CREB and AKT, which are related to oxidative capacity. Resistance training increased PGC-1α (~59%) and TFAM (~48%) expression compared to the Ovariectomy-Sedentary group. The combination of resistance training and estradiol replacement was superior to the ovariectomy-sedentary and ovariectomy-resistance training treatments regarding the gastrocnemius muscle. Estrogen deficiency altered the expression of genes and proteins that favour the development of a mitochondrial dysfunction phenotype, which was improved with resistance training and was partially improved by estradiol replacement.

Insulin-like growth factor binding proteins in follicular fluid from morphologically distinct healthy and atretic bovine antral follicles.

In bovine follicles 2-5 mm in diameter, two morphologically distinct types of healthy follicles and two types of atretic follicles have been described recently. Healthy follicles either have columnar basal granulosa cells with follicular basal lamina composed of many layers or 'loops' or they have rounded basal cells with a conventional single-layered, aligned follicular basal lamina. In atretic follicles, cell death either commences at the basal layer and progresses to the antrum (basal atresia) with macrophage penetration of the membrana granulosa or death progresses from the antrum in a basal direction (antral atresia). Little is known about how these different phenotypes develop. To determine whether insulin-like growth factor binding protein (IGFBP) levels in follicular fluid differ between these different types of follicles, we measured IGFBP levels in fluids from these follicles. A total of 61 follicles were assessed by light microscopy and characterized by morphological analysis as either healthy, with columnar or rounded basal granulosa cells, or as undergoing antral or basal atresia. The IGFBP concentration in the follicular fluid of individual follicles from the four groups (n = 12-20 per group) was identified by Western ligand blots using (125)I-insulin-like growth factor (IGF)-II as a probe. Insulin-like growth factor binding proteins 2, 3 (44 and 40 kDa), 4 (glycosylated and non-glycosylated) and 5 were observed. The levels (per volume of fluid) of IGFBPs 2, 4 and 5 were greater in atretic follicles than in healthy follicles. However, there were no statistical differences in levels of each IGFBP between either the two types of healthy follicle or between the two types of atretic follicles. Thus, IGFBP levels are not related to the different types of healthy or atretic follicles.

Quantitative analysis of human immunodeficiency virus type 1 antibody reactivity by western immunoblots: evaluation of relative antibody levels in seropositive individuals and mothers.

A quantitative analysis of antibody responses to human immunodeficiency virus type 1 (HIV-1) proteins using Western immunoblots and 125I-labeled protein A is reproducible and can be validated. The antibody levels obtained by Western immunoblots were compared with stoichiometric p24 radioimmunoassay over a wide range of antibody (correlation coefficient, .94; P less than .001). Antibody levels to gp160 and gp120 were validated using purified antigens. Analysis of antibody levels from 31 seropositive individuals revealed a statistically significant correlation between antibody levels to p24 and the other viral proteins except gp120. Anti-gag p24 antibody was strongly correlated with antibodies to other env products, specifically gp41 and gp160. Using the validated assay, HIV-1-infected mothers of infants were found to have highly variable levels of antibody to all viral proteins. Mothers of infected infants did not differ significantly from mothers of uninfected infants in antibody pattern or levels to any viral protein including gp120.