Genotoxic evaluation of the antimalarial drugs artemisinin and artesunate in human HepG2 cells and effects on CASP3 and SOD1 gene expressions

The malaria treatment recommended by the World Health Organization involves medicines derived from artemisinin, an active compound extracted from the plant Artemisia annua, and some of its derivatives, such as artesunate. Considering the lack of data regarding the genotoxic effects of these compounds in human cells, the objective of this study was to evaluate the cytotoxicity and genotoxicity, and expressions of the CASP3 and SOD1 genes in a cultured human hepatocellular liver carcinoma cell line (HepG2 cells) treated with artemisinin and artesunate. We tested concentrations of 2.5, 5, 7.5, 10, and 20 μg/mL of both substances with a resazurin cytotoxicity assay, and the concentrations used in the genotoxicity experiments (2.5, 5, and 10 μg/mL) and gene expression analysis (5 mg/mL) were determined. The results of the comet assay in cells treated with artemisinin and artesunate showed a significant dosedependent increase (P < 0.001) in the number of cells with DNA damage at all concentrations tested. However, the gene expression analysis revealed no significant change in expression of CASP3 or SOD1. Our data showed that although artemisinin and artesunate exhibited genotoxic effects in cultured HepG2 cells, they did not significantly alter expression of the CASP3 and SOD1 genes at the doses tested. ©FUNPEC-RP.

Meiotic pairing of B chromosomes, multiple sexual system, and Robertsonian fusion in the red brocket deer Mazama americana (Mammalia, Cervidae)

Deer species of the genus Mazama show significant inter and intraspecific chromosomal variation due to the occurrence of rearrangements and B chromosomes. Given that carriers of aneuploidies and structural rearrangements often show anomalous chromosome pairings, we here performed a synaptonemal complex analysis to study chromosome pairing behavior in a red brocket deer (Mazama americana) individual that is heterozygous for a Robertsonian translocation, is a B chromosome carrier, and has a multiple sex chromosome system (XY1Y2). The synaptonemal complex in spermatocytes showed normal chromosome pairings for all chromosomes, including the autosomal and sex trivalents. The electromicrographs showed homology among B chromosomes since they formed bivalents, but they also appeared as univalents, indicating their anomalous behavior and non-Mendelian segregation. Thus, synaptonemal complex analysis is a useful tool to evaluate the role of B chromosomes and rearrangements during meiosis on the intraspecific chromosomal variation that is observed in the majority of Mazama species. © FUNPEC-RP.

Chromosomal characteristics of a Brazilian whip spider (Amblypygi) and evolutionary relationships with other arachnid orders

We analyzed mitotic and meiotic cells of a Brazilian amblypygid, Heterophrynus longicornis, using conventional and molecular cytogenetic techniques (Giemsa staining, C-banding, Ag-NOR, and FISH with rDNA probe). This is the first study that focuses solely on amblypygid chromosomes; it was undertaken to add data on cytogenetic knowledge of this group and contribute to the understanding of chromosome evolution in the Arachnida. We found 2n = 66 for male and female individuals, monocentric chromosomes, and absence of morphologically differentiated sex chromosomes. C-banding showed heterochromatin in the pericentromeric region of most chromosomes. Mitotic and meiotic nuclei submitted to silver impregnation and FISH revealed, respectively, Ag-NORs and ribosomal genes in the terminal region of two chromosome pairs. Most chromosome features that we observed in H. longicornis are shared with species of other arachnid orders; however, the absence of morphologically differentiated sex chromosomes in amblypygid contrasts with the remarkable variety of sex chromosome systems recorded for the Araneae. Consequently, we conclude that analysis of species of the Tetrapulmonata clade is useful for understanding the trends of sex chromosome evolution in this arachnid group. © FUNPEC-RP.

Genetic structure of Aedes aegypti populations determined using pairwise comparisons

The biological characteristics of Aedes aegypti (Diptera, Culicidae), which is a vector of dengue and yellow fever, make this organism a good model for studying population structure and the events that may influence it under the effect of human activity. We assessed the genetic variability of five A. aegypti populations using RAPD-PCR technique and six primers. Four populations were from Brazil and one was from the USA. A total of 165 polymorphic DNA loci were generated. Considering the six primers and the five populations, the mean value of inter-population genetic diversity (Gst) was 0.277, which is considered high according to the Wright classification. However, pairwise comparisons of the populations gave variable Gst values ranging from 0.044 to 0.289. This variation followed the population's geographic distance to some extent but was also influenced by human activity. The lowest Gst values were obtained in the comparison of populations from cities with intensive commercial and medical contacts. These mosquito populations were previously classified as insecticide resistant, susceptible, or with decreased susceptibility; this parameter apparently had an effect on the Gst values obtained in the pairwise comparisons. ©FUNPEC-RP.

Effect of glycerol on GHR and IGF-1 gene expression in breast muscle and on growth of Japanese meat quails

We evaluated messenger RNA (mRNA) expression of the growth-hormone (GHR) and insulin-like growth factor (IGF-1) genes in 28-day-old Japanese meat quails fed diets containing 0, 8, or 12% dietary glycerol in substitution of corn. Total RNA was extracted from the breast muscle and the DNA was amplified with specific primers using real-time PCR. Feed conversion ratio and feed intake were evaluated. The birds fed 8 and 12% glycerol presented higher IGF-1 mRNA expression [0.059 and 0.049 arbitrary units (AU), respectively] relative to those not fed with glycerol (0.029 AU), while 12% glycerol reduced GHR mRNA expression (0.022 AU). Dietary inclusion of 8% glycerol promoted similar performance results (feed conversion) as the diet with no glycerol. We conclude that inclusion of glycerol in the diet affects GHR and IGF-1 gene expression in Japanese meat quails. However, considering the performance results and the expression of the GHR and IGF-1 genes, 8% glycerol may be safely included in the diet of meat quails. © FUNPEC-RP.

Bovine herpesvirus type 5 infection regulates Bax/BCL-2 ratio

Bovine herpesvirus 5 (BoHV-5) is an α-herpesvirus that causes neurological disease in young cattle and is also occasionally involved in reproductive disorders. Although there have been many studies of the apoptotic pathways induced by viruses belonging to the family Herpesviridae, there is little information about the intrinsic programmed cell death pathway in host-BoHV-5 interactions. We found that BoHV-5 is able to replicate in both mesenchymal and epithelial cell lines, provoking cytopathology that is characterized by cellular swelling and cell fusion. Viral antigens were detected in infected cells by immunofluorescence assay at 48 to 96 h post-infection (p.i.). At 48 to 72 h p.i., anti-apoptotic BCL-2 antigens were found at higher levels than Bax antigens; the latter is considered a pro-apoptotic protein. Infected cells had increased BCL-2 phenotype cells from 48 to 96 h p.i., based on flow cytometric analysis. At 48 to 96 h p.i., Bax mRNA was not expressed in any of the infected cell monolayers. In contrast, BCL-2 mRNA was found at high levels at all p.i. in both types of cells. BoHV-5 replication apparently modulates BCL-2 expression and gene transcription, enhancing production of virus progeny. © FUNPEC-RP.

Assessment of clinical scoring systems for the diagnosis of Williams-Beuren syndrome

Williams-Beuren syndrome (WBS) is a genetic disorder characterized by physical and intellectual developmental delay, associated with congenital heart disease and facial dysmorphism. WBS is caused by a microdeletion on chromosome 7 (7q11.23), which encompasses the elastin (ELN) gene and about 27 other genes. The gold standard for WBS laboratory diagnosis is FISH (fluorescence in situ hybridization), which is very costly. As a possible alternative, we investigated the accuracy of three clinical diagnostic scoring systems in 250 patients with WBS diagnosed by FISH. We concluded that all three systems could be used for the clinical diagnosis of WBS, but they all gave a low percentage of false-positive (6.0-9.2%) and false-negative (0.8-4.0%) results. Therefore, their use should be associated with FISH testing. © FUNPEC-RP.

Comparison of factor-analytic and reduced rank models for test-day milk yield in Gyr dairy cattle (Bos indicus)

We analyzed 46,161 monthly test-day records of milk production from 7453 first lactations of crossbred dairy Gyr (Bos indicus) x Holstein cows. The following seven models were compared: standard multivariate model (M10), three reduced rank models fitting the first 2, 3, or 4 genetic principal components, and three models considering a 2-, 3-, or 4-factor structure for the genetic covariance matrix. Full rank residual covariance matrices were considered for all models. The model fitting the first two principal components (PC2) was the best according to the model selection criteria. Similar phenotypic, genetic, and residual variances were obtained with models M10 and PC2. The heritability estimates ranged from 0.14 to 0.21 and from 0.13 to 0.21 for models M10 and PC2, respectively. The genetic correlations obtained with model PC2 were slightly higher than those estimated with model M10. PC2 markedly reduced the number of parameters estimated and the time spent to reach convergence. We concluded that two principal components are sufficient to model the structure of genetic covariances between test-day milk yields. © FUNPEC-RP.

An HPLC-UV method for the quantification of zidovudine in rat plasma

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Associações entre nível, oferta de atividade física no trabalho e atividade física de lazer

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influência da adição da espécie Acidithiobacillus ferrooxidans e de íons Cl- na dissolução da calcopirita (CuFeS'IND2')

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Metilação e expressão do gene BRCA1 em meningiomas

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

Funções de pedotransferência para a curva de resistência do solo à penetração

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Desenvolvimento e caracterização físico-química de sistemas líquido cristalinos contendo palmitato de retinol: quantificação química, avaliação da atividade antioxidante, estudos de liberação e bioadesão in vitro

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Variação sexual, ontogenética e ambiental do veneno de Bothrops jararaca da icro-região de Botucatu-SP: caracterização enzimática, bioquímica e farmacológica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)