Forensic characterization of Brazilian regional populations through massive parallel sequencing of 124 SNPs included in HID ion Ampliseq Identity Panel.

Use of Massive Parallel Sequencing (MPS) techniques has been investigated by forensic community aiming introduction of such methods in routine forensic casework analyses. Interesting features presented by MPS include high-throughput, ability to simultaneous genotyping of significant number of samples and forensic markers, workflow automation, among others. Emergence of single nucleotide polymorphism (SNP) as forensic relevant markers was facilitated in this process, since concurrent typing of larger marker sets is necessary for obtaining same levels of individual discrimination provided by other marker categories. In this context, HID Ion Ampliseq Identity Panel is a commercial solution with forensic purposes comprising simultaneous analysis of 90 highly informative autosomal SNPs and 34 Y -chromosome superior clade SNPs for male lineage haplotyping. SNP typing can be obtained with smaller amplicons, and this panel was designed for efficient processing of critical or challenging forensic samples. In this work, a sample of 432 individuals from all five Brazilian geopolitical regions was evaluated with this panel, in order to access feasibility of this panel use in a national basis. Results obtained for all five regions, including forensic parameters, show that this marker set can be efficiently employed for Brazilian nationals in human identification or kinship determination applications, due to high levels of genetic discriminative information content displayed by Brazilians. Interpopulation comparison studies were executed among Brazilian regional populations and 26 worldwide populations, in order to access genetic stratification occurrence. Some levels of population structure were identified, and impact on database design was discussed. Y-chromosome haplotyping of Brazilian samples revealed high levels of European ancestry in Brazilian male lineages, and utility of haplotyping in real forensic casework is addressed. Finally, genotyping and sequencing efficiency with this panel were addressed, as an effort to appraise the adequacy of this panel use in Brazilian national forensic demands.

Dehydroepiandrosterone sulphate enhances IgG and Interferon-gamma production during immunization to tuberculosis in young but not aged mice.

Ageing of the endocrine system (endocrinosenescence) has been closely related to immunosenescence. Dehydroepiandrosterone sulphate (DHEAS), a steroid hormone produced by the adrenals with reported enhancing immunomodulatory properties, consistently decline during ageing in parallel to detrimental increase in peripheral glucocorticoids. We investigated here the adjuvant effects of DHEAS during intraperitoneal immunization to Mycobacterium tuberculosis heat shock protein 70 (mycHSP70) in old (24 months) as well as young (3 months) BALB/c mice. Both young and old mice had significantly higher Immunoglobulin G (IgG) levels following immunization. Young mice co-immunized with mycHSP70-DHEAS presented an early increase in specific IgG levels and showed increased Interferon-gamma production compared to old mice. Also, T cells of immunized young animals were consistently more resistant to the immunosuppressive effects of glucocorticoids and to DHEAS. DHEAS was not effective in modulating antigen-specific T-cell proliferation, Interleukin-2 production or percentage of recent activated T-cell subsets (CD4 + CD69 + and CD8 + CD69 +). Our data further indicate mycHSP70 as a putative good antigen in vaccine to tuberculosis. Our data also suggest that DHEAS produced adjuvant effects upon humoral and some cellular immune responses of young, but not old mice and indicate that immunization with DHEAS is capable of changing T-cell responses to steroids.

14-3-3 gene characterization and description of a second 14-3-3 isoform in both Echinococcus granulosus and E. multilocularis.

Members of the 14-3-3 protein family have been identified as regulatory molecules in intracellular signaling pathways and cell cycle control. Previously, the first Echinococcus 14-3-3 isoform (E14-3-3.1) was isolated from E. granulosus and E. multilocularis metacestode stages. Hyperexpression of this isoform was claimed to be associated with non-restricted tumor-like growth of the E. multilocularis metacestode. In this report, we describe the characterization of a 14-3-3 cDNA from E. granulosus and E. multilocularis corresponding to a second isoform of this family, E14-3-3.2. The characterized 14-3-3 gene was interrupted by two introns whose sequence and positions were conserved in both Echinococcus species. The deduced amino acid sequence of E14-3-3.2 showed 88% identity to the E14-3-3.1 isoform and 52% identity to a third Echinococcus isoform (E14-3-3.3) described by other authors. These findings, coupled to Southern blot analysis, suggest the presence of more than one 14-3-3 gene in Echinococcus. Phylogenenetically, the Echinococcus 14-3-3.1 and 14-3-3.2 isoforms appeared to cluster with zeta-type ("pro-tumorigenic") 14-3-3 isoforms from closely related organisms, whereas the E14-3-3.3 isoform grouped with 14-3-3 epsilon isoforms. The presence of more than one 14-3-3 isoform might indicate isoform-specific roles in the different parasite stages of Echinococcus.