67 research outputs found
FYB polymorphisms in Brazilian patients with type I diabetes mellitus and autoimmune polyglandular syndrome type III
The aim of this study was to perform an association study between seven Fyn-binding protein gene (FYB)-tag single nucleotide polymorphisms (SNPs) and type I diabetes mellitus (T1DM), as well as with disease age of onset. We also assessed the role of FYB SNPs in the insurgence of autoimmune polyglandular syndrome type III (APSIII), characterized by the simultaneous presence of autoimmune thyroid disease and celiac disease, in patients with T1DM from a Northeastern Brazilian population. One hundred and seventy-seven patients with T1DM and 190 healthy individuals were genotyped for seven tag SNPs, covering most of the FYB locus, using real-time polymerase chain reaction amplification. There was no significant difference in the distribution of allele and genotype frequencies among patients and healthy individuals. Moreover, none of the tag SNPs were associated either to T1DM age of onset or to the insurgence of APSIII. However, since the FYB protein is a key component in T cell response, its gene variants might play a role in protein function, which might be testable in a population with different genetic backgrounds or by using functional assays
A rapid screening of ancestry for genetic association studies in an admixed population from Pernambuco, Brazil
Genetic association studies determine how genes influence traits. However, non-detected population substructure may bias the analysis, resulting in spurious results. One method to detect substructure is to genotype ancestry informative markers (AIMs) besides the candidate variants, quantifying how much ancestral populations contribute to the samples' genetic background. The present study aimed to use a minimum quantity of markers, while retaining full potential to estimate ancestries. We tested the feasibility of a subset of the 12 most informative markers from a previously established study to estimate influence from three ancestral populations: European, African and Amerindian. The results showed that in a sample with a diverse ethnicity (N = 822) derived from 1000 Genomes database, the 12 AIMs had the same capacity to estimate ancestries when compared to the original set of 128 AIMs, since estimates from the two panels were closely correlated. Thus, these 12 SNPs were used to estimate ancestry in a new sample (N = 192) from an admixed population in Recife, Northeast Brazil. The ancestry estimates from Recife subjects were in accordance with previous studies, showing that Northeastern Brazilian populations show great influence from European ancestry (59.7%), followed by African (23.0%) and Amerindian (17.3%) ancestries. Ethnicity self-classification according to skin-color was confirmed to be a poor indicator of population substructure in Brazilians, since ancestry estimates overlapped between classifications. Thus, our streamlined panel of 12 markers may substitute panels with more markers, while retaining the capacity to control for population substructure and admixture, thereby reducing sample processing time
Postmenopausal Osteoporosis reference genes for qPCR expression assays
Osteoporosis (OP) is a multifactorial disease influenced by genetic factors in more than half of the cases. In spite of the efforts to clarify the relationship among genetic factors and susceptibility to develop OP, many genetic associations need to be further functionally validated. Besides, some limitations as the choice of stably expressed reference genes (RG) should be overcome to ensure the quality and reproducibility of gene expression assays. To our knowledge, a validation study for RG in OP is still missing. We compared the expression levels, using polymerase chain reaction quantitative real time (qPCR) of 10 RG (G6PD, B2M, GUSB, HSP90, EF1A, RPLP0, GAPDH, ACTB, 18 S and HPRT1) to assess their suitability in OP analysis by using GeNorm, Normfinder, BestKeeper and RefFinder programs. A minimal number of two RG was recommended by GeNorm to obtain a reliable normalization. RPLP0 and B2M were identified as the most stable genes in OP studies while ACTB, 18 S and HPRT1 were inadequate for normalization in our data set. Moreover, we showed the dramatic effects of suboptimal RG choice on the quantification of a target gene, highlighting the importance in the identification of the most appropriate reference gene to specific diseases. We suggest the use of RPLP0 and B2M as the most stable reference genes while we do not recommend the use of the least stable reference genes HPRT1, 18 S and ACTB in OP expression assays using PBMC as biological source. Additionally, we emphasize the importance of individualized and careful choice in software and reference genes selection
CaracterizaĆ§Ć£o do perfil comportamental e de competĆŖncia social de indivĆduos com a sĆndrome del22q11.2
Deletions and duplications of the 22q11.2 region in spermatozoa from DiGeorge/velocardiofacial fathers
Alterations in gene expression profiles correlated with cisplatin cytotoxicity in the glioma U343 cell line
Gliomas are the most common tumors in the central nervous system, the average survival time of patients with glioblastoma multiforme being about 1 year from diagnosis, in spite of harsh therapy. Aiming to study the transcriptional profiles displayed by glioma cells undergoing cisplatin treatment, gene expression analysis was performed by the cDNA microarray method. Cell survival and apoptosis induction following treatment were also evaluated. Drug concentrations of 12.5 to 300 Ī¼M caused a pronounced reduction in cell survival rates five days after treatment, whereas concentrations higher than 25 Ī¼M were effective in reducing the survival rates to ~1%. However, the maximum apoptosis frequency was 20.4% for 25 Ī¼M cisplatin in cells analyzed at 72 h, indicating that apoptosis is not the only kind of cell death induced by cisplatin. An analysis of gene expression revealed 67 significantly (FDR < 0.05) modulated genes: 29 of which down- and 38 up-regulated. These genes belong to several classes (metabolism, protein localization, cell proliferation, apoptosis, adhesion, stress response, cell cycle and DNA repair) that may represent several affected cell processes under the influence of cisplatin treatment. The expression pattern of three genes (RHOA, LIMK2 and TIMP2) was confirmed by the real time PCR method
Receptor Complementation and Mutagenesis Reveal SR-BI as an Essential HCV Entry Factor and Functionally Imply Its Intra- and Extra-Cellular Domains
HCV entry into cells is a multi-step and slow process. It is believed that the
initial capture of HCV particles by glycosaminoglycans and/or lipoprotein
receptors is followed by coordinated interactions with the scavenger receptor
class B type I (SR-BI), a major receptor of high-density lipoprotein (HDL), the
CD81 tetraspanin, and the tight junction protein Claudin-1, ultimately leading
to uptake and cellular penetration of HCV via low-pH endosomes.
Several reports have indicated that HDL promotes HCV entry through interaction
with SR-BI. This pathway remains largely elusive, although it was shown that HDL
neither associates with HCV particles nor modulates HCV binding to SR-BI. In
contrast to CD81 and Claudin-1, the importance of SR-BI has only been addressed
indirectly because of lack of cells in which functional complementation assays
with mutant receptors could be performed. Here we identified for the first time
two cell types that supported HCVpp and HCVcc entry upon ectopic SR-BI
expression. Remarkably, the undetectable expression of SR-BI in rat hepatoma
cells allowed unambiguous investigation of human SR-BI functions during HCV
entry. By expressing different SR-BI mutants in either cell line, our results
revealed features of SR-BI intracellular domains that influence HCV infectivity
without affecting receptor binding and stimulation of HCV entry induced by
HDL/SR-BI interaction. Conversely, we identified positions of SR-BI ectodomain
that, by altering HCV binding, inhibit entry. Finally, we characterized
alternative ectodomain determinants that, by reducing SR-BI cholesterol uptake
and efflux functions, abolish HDL-mediated infection-enhancement. Altogether, we
demonstrate that SR-BI is an essential HCV entry factor. Moreover, our results
highlight specific SR-BI determinants required during HCV entry and
physiological lipid transfer functions hijacked by HCV to favor infection
Assessing the human immune system through blood transcriptomics
Blood is the pipeline of the immune system. Assessing changes in transcript abundance in blood on a genome-wide scale affords a comprehensive view of the status of the immune system in health and disease. This review summarizes the work that has used this approach to identify therapeutic targets and biomarker signatures in the field of autoimmunity and infectious disease. Recent technological and methodological advances that will carry the blood transcriptome research field forward are also discussed
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