Identification of Suitable Reference Genes for Real Time Quantitative Polymerase Chain Reaction Assays on Pectoralis major Muscle in Chicken (Gallus gallus ).

Thirteen reference genes were investigated to determine their stability to be used as a housekeeping in gene expression studies in skeletal muscle of chickens. Five different algorithms were used for ranking of reference genes and results suggested that individual rankings of the genes differed among them. The stability of the expression of reference genes were validated using samples obtained from the Pectoralis major muscle in chicken. Samples were obtained from chickens in different development periods post hatch and under different nutritional diets. For gene expression calculation the ΔΔCt approach was applied to compare relative expression of pairs of genes within each of 52 samples when normalized to mitochondrially encoded cytochrome c oxidase II (MT-CO2) target gene. Our findings showed that hydroxymethylbilane synthase (HMBS) and hypoxanthine phosphoribosyl transferase 1 (HPRT1) are the most stable reference genes while transferrin receptor (TFRC) and beta-2-microglobulin (B2M) ranked as the least stable genes in the Pectoralis major muscle of chickens. Moreover, our results revealed that HMBS and HPRT1 gene expression did not change due to dietary variations and thus it is recommended for accurate normalization of RT-qPCR data in chicken Pectoralis major muscle.

Multilocus enzyme electrophoresis analysis and exoenzymatic activity of Candida albicans strains isolated from women with vaginal candidiasis.

Twenty-eight Candida albicans strains obtained from women with vaginal candidiasis were tested for phospholipase and proteinase production and clustered by multilocus enzyme electrophoresis (MLEE). The proteolytic and phospholipidic activity were considered moderate (0.56 ± 0.12 mm and 0.53 ± 0.09 mm, respectively) for all isolates. The isoenzymes malate dehydrogenase (MDH) and sorbitol dehydrogenase (SDH) showed strong intra-specific discriminatory power. The numerical and genetic interpretation of the bands produced by the isoenzymes tested presented similar discriminatory power. The genetic diversity of the isolates was measured by allelic and genic frequency, perceptual index of polymorphic loci (P = 87.5%), average number of alleles per locus, average number of alleles per polymorphic locus, average heterozygosity observed and average heterozygosity expected. We verified that three isoenzymatic loci (Adh, Gdh and Sdh-2) were not in Hardy-Weinberg equilibrium. A dendrogram constructed based on the genetic distance matrix of Nei showed seven clusters; 57.15% (16) of the isolates were considered highly related or indistinguishable, and 42.85% were considered moderately related or unrelated. We did not find a relationship between the clusters and the exoenzymes production.