ABSTRACT The objectives of this study were to genotype single nucleotide polymorphisms (SNP) AF159246:g.2959A>G (CAST/DdeI) and AF248054.2:g.6545C>T (CAPN4751) in beef cattle by PCR-RFLP (Polymerase Chain Reaction - Restriction Fragment Length Polymorphism), using the restriction enzyme DdeI for both SNP, and describe the use of these genotyping methodologies for the first time. For the SNP located in the CAST gene, new primers were designed, and for the SNP of the CAPN1 gene, the same primers previously described in the literature were used. Bonsmara, Caracu, Senepol, Nelore, and Angus bulls were chosen from among the most used bulls in breeding programs according to their genealogy and the lowest possible degree of parentage between them to ensure an experimental sample representative of the genetic variability in each breed. For the CAST and CAPN1 genes, respectively, the following number of animals were analyzed: Bonsmara (n = 25/22), Caracu (n = 25/26), Senepol (n = 25/24), Nelore (n = 26/26), and Angus (n = 25/24). The accuracy of these methodologies was confirmed by direct sequencing of PCR products generated for the two polymorphisms. The new primers developed for CAST/DdeI SNP detection and the use of DdeI enzyme for CAPN4751 SNP detection were effective in genotyping, since no inconclusive genotypes were observed for these genes. Thus, the genotyping of beef cattle using the PCR-RFLP technique for CAST and CAPN1 genes is robust, relatively inexpensive, and easy to perform in any basic molecular biology laboratory. If the association of these markers with traits of economic interest in beef cattle is confirmed in new studies, these methodologies may contribute to the selection of animals with superior genetics, i.e., with the potential to produce better-quality meat, either by marker-assisted selection or by the inclusion of these polymorphisms in high-density marker panels.

ABSTRACT Due to the severity of chronic hepatitis C, there are multiple factors that can negatively affect the quality of life of infected patients. The aim of this study was to evaluate changes in the health-related quality of life (HRQoL) in patients under second-generation direct-acting antiviral (DAA) (interferon-free) therapies and to assess treatment effectiveness. This was an observational study conducted in Curitiba (Brazil) using two instruments (a generic and a specific) for measuring the quality of life in patients with chronic hepatitis C, the Short Form-36 (SF-36) and the Chronic Liver Disease Questionnaire (CLDQ) for liver disease evaluation. The study included patients receiving any interferon-free therapy for hepatitis C treatment during 2016 and 2017. Data were collected before, during, and after treatment regarding the two questionnaires, effectiveness and safety. Fifty-six patients fulfilled all eligibility criteria and were included for analysis. Sustained virological response was obtained in 88% of the patients. They were mainly genotype 1, cirrhotic and treated with sofosbuvir combined with daclatasvir or sofosbuvir with simeprevir. Improvement in the quality of life was observed for several domains in both questionnaires (p < 0.05) in the comparison before and after treatment. Patients receiving sofosbuvir with daclatasvir had significantly lower scores compared to the group receiving sofosbuvir with simeprevir. Second-generation DAA therapies were effective and have considerably increased the HRQoL of patients with chronic hepatitis C virus.

The objective of the present study was to estimate the allelic and genotypic frequencies of the polymorphisms E2FB (AY138588.1: c.305C> T), located in the leptin gene (LEP), and TG5 (X05380.1:g.-422C>T), located in the thyroglobulin gene (TG), and evaluate the association of these polymorphisms in crossbred cattle of seven distinct genetic groups with the following traits: slaughter weight (SW), hot carcass weight (HCW), hot carcass yield (HCY), carcass fat thickness (CFT), ribeye area (REA), marbling (MARM) and shear force (SF). The animals were genotyped using the PCR-RFLP (Polymorphism Chain Reaction-Restriction Fragment Length Polymorphism) technique, using 201 products obtained from F1 Caracu × Nellore, Angus × Nellore and Valdostana × Nellore cows, mated to Canchim, Caracu and Red Angus bulls (only Caracu × Nellore cows were used with Red Angus bulls). The allelic and genotypic frequencies were compared using the Chi-squared test. Associations between the genotype of each polymorphism and the traits were analyzed using the General Linear Model (GLM) of statistical software SAS. The least squares means of genotypes of the polymorphisms were compared using Student's t test. The E2FB polymorphism in the LEP gene was associated with CFT, showing the potential for use in national programs for genetic improvement of beef cattle, through the inclusion of SNP in genotyping commercial tests. The TG5 polymorphism in the TG gene was not associated with any of the evaluated traits and was considered ineffective for selection of beef cattle in Brazilian herds.