Inga edulis fruits: a new source of bioactive anthocyanins.

The extraction conditions and chromatographic analysis from seeds of Inga edulis were optimized and provided one anthocyanin from aqueous fraction and a mixture of three anthocyanins from methanolic fraction. The pure anthocyanin obtained was subjected to structural modifications and the products obtained were subjected to chemical and pharmacological assays, as well as quantum chemical calculations based on DFT and TD-DFT methods. Hence, the anthocyanin fractions were evaluated for their chemical-pharmacological potential through chemical and biological assays: antioxidant activity by the DPPH, determination of the Solar Protection Factor (SPF) and cytotoxic activity (hepatocellular carcinoma infected with hepatitis C virus). The results indicated that even the anthocyanin and derivatized compounds having high antioxidant potential showed an SPF lower than six, which is lower than the minimum accepted by current Brazilian legislation. In addition, none of compounds presented significant cytotoxic activity against the tumour cell line studied.

Polysiloxane-poly(propylene oxide) hybrid discs as solid phase in anti-HCV detection using a recombinant core protein.

In this work, siloxane-poly(propylene oxide) discs (PPO disc) prepared using the sol-gel process were used as solid phase in enzyme-linked immunosorbent assays (ELISA) for the detection of anti-hepatitis C virus (HCV) antibodies. The HCV RNA from serum (genotype 1b) was submitted to the RT-PCR technique and subsequent amplification of the HCV core 408 pb. This fragment was cloned into expression vector pET42a and expressed in Escherichia coli as recombinant protein with glutathione S-transferase (GST). Cell cultures were grown and induced having a final concentration of 0.4 x 10(-3) mol L(-1) of IPTG. After induction, the cells were harvested and the soluble fraction was analyzed using polyacrilamide gel 15% showing a band with an approximate molecular weight of 44 kDa, the expected size for this GST-fused recombinant protein. The recombinant protein was purified and confirmed by immunological detection using HCV-positive serum and showed no cross-reactivity with positive samples for other infectious diseases. An ELISA was established using 1.25 ng of recombinant protein per PPO disc, a dilution of 1:10,000 and 1:40 for a peroxidase conjugate and serum, respectively, and solutions of hydrogen peroxide and 3,3',5,5'-tetra-methylbenzidine in a ratio of 1:1. The proposed methodology was compared with the ELISA conventional polystyrene-plate procedure and the performance of the PPO discs as a matrix for immunodetection gave an easy synthesis, good performance and reproducibility for commercial application.

Doença arterial coronariana: um novo paradigma na AIDS / Coronary artery disease: a new paradigm in AIDS

A inclusão de indivíduos portadores da infecção pelo HIV num grupo propenso às altraçòes metabólicas, fortalece a hipótese de que a AIDS contribui como um fator de risco para o desenvolvimento de desordens vaso-oclusivas. O curso e a progressão da AIDS, bem como as medidas terapêuticas contra o HIV têm sido capazes de mostrar uma infinidade de alterações metabólicas aos quais os portadores estão sujeitos. Esses transtornos afetam o metabolismo de componentes plasmáticos como os lipídeos e a homocisteína e tem sido verificado nos portadores do HIV como consequênciade três fatores predominantemente: (i) a própria infecção viral per se que altera os níveis de lipídeos plasmáticos; (ii) deficiências vitamínicas e de micronutrientes, favorecendo a hiper-homocisteinemia (iii) a terapia medicamentosa antiretroviral, que acompanha efeitos idiossincráticos no metabolismo lipídico do portador. Nesse contexto, o indivíduo infectado pelo HIV está inserido num rol de anormalidades que são fatores de risco para a aterogênese. Essas observações podem ser valiosas quando se verifica que, se a sobrevida do portador está sendo aumentada por ocasião da terapia mais efetiva, é por outro lado possível, que seu risco de desenvolver alteraçòes metabólicas e, portanto, iniciar um processo aterogênico, ou exacerbar algum preexistente, tenha também aumento proporcional

Fator de crescimento derivado de plaquetas humanas obtido por uma metodologia rápida e de baixo custo / Human platelet-derived growth factor obtained by means of a fast and inexpensive methodology

O fator de crescimento derivado de plaquetas (PDGF) é uma proteína catiônica armazenado principalmente nos grânulos-alfa plaquetário. O PDGF por ser altamente mitogênico, principalmente para fibroblastos e leiomiócitos, é de grande importância no processo de regeneração tecidual. Neste trabalho, o PDGF foi obtido a partir do lisado de plaquetas humanas (5 x 10 plaquetas/mL) vencidas, purificado por cromatografia de troca iônica em resina de CM-Sepharose. A proteína eluída na fração catiônica, foi identificada por anticorpos policlonais anti-PDGF (AA e BB) e, posteriormente, sua atividade mitogênica confirmada pela incorporação da [H3]-TdR usando fibroblastos da linhagem celular NIH/3T3 em cultura. A comparação entre os efeitos mitogênicos sobre o PDGF, fração catiônica, com uma proteína recombinante controle (PDGF-AB), revelou que a proteína parcialmente purificada induziu semelhante estimulação, 157.567 cpm e 165.796 cpm, respectivamente. Os resultados obtidos evidenciam a aplicabilidade dessas condições experimentais na obtenção do PDGF, preservando sua atividade biológica, através de procedimentos de baixa complexibilidade, eficiente e de custo operacional reduzido, possibilitando seu uso em experimentos de regeneração tecidual

CCR5 genotype and plasma ß-chemokine concentration of Brazilian HIV-infected individuals

The 32-bp deletion in the HIV-1 co-receptor CCR5 confers a high degree of resistance to HIV-1 infection in homozygous individuals for the deleted allele and partial protection against HIV-1 during disease progression in heterozygotes. Natural ligands for CCR5, MIP-1alpha, MIP-1ß and RANTES, have been shown to inhibit HIV replication in CD4+ T cells. In the present study, we examined the CCR5 genotype by PCR and the plasma levels of RANTES and MIP-1alpha by ELISA among blood donors (N = 26) and among HIV-1-infected individuals (N = 129). The control group consisted of healthy adult volunteers and HIV-1-infected subjects were an asymptomatic and heterogeneous group of individuals with regard to immunologic and virologic markers of HIV-1 disease. The frequency of the CCR5 mutant allele (delta32ccr5) in this population was 0.032; however, no delta32ccr5 homozygote was detected. These results could be related to the intense ethnic admixture of the Brazilian population. There was no correlation between circulating ß-chemokines (MIP-1alpha, RANTES) and viral load in HIV-infected individuals. RANTES concentrations in plasma samples from HIV+ patients carrying the homozygous CCR5 allele (CCR5/CCR5) (28.23 ng/ml) were higher than in the control samples (16.07 ng/ml; P<0.05); however, this HIV+ patient group (mean 26.23 pg/ml) had significantly lower concentrations of MIP-1alpha than those observed in control samples (mean 31.20 pg/ml; P<0.05). Both HIV-1-infected and uninfected individuals heterozygous for the delta32ccr5 allele had significantly lower concentrations of circulating RANTES (mean 16.07 and 6.11 ng/ml, respectively) than CCR5/CCR5 individuals (mean 28.23 and 16.07 ng/ml, respectively; P<0.05). These findings suggest that the CCR5 allele and ß-chemokine production may affect the immunopathogenesis of HIV-1

Rattlesnake venom: action upon erythrocytes and leucocytes of rats

Hemos estudiado la acción del veneno completo de serpiente de cascabel sobre los glóbulos rojos y leucocitos de ratas adultas machos y hembras. Los animales fueron quirúrgicamente canulados para realizar recolecciones de sangre directamente de la vena cava inferior, e inyectados intramuscularmente en el muslo con la mezcla de veneno obtenido de un gran número de cascbeles. Los signos mostrados por los animales fueron parálisis de la parte posterior del cuerpo, déficit de la coordinación motora y dificultad repiratoria, con muerte en algunos casos. La autopsia mostró hemorragias petequiales en el intestino y yeyuno, además de oscurecimiento visceral y obstrucción de los vasos sanguíneos verificada en el examen histopatológico. Los exámenes sanguíneos mostrarón cambios de color hacia marrón oscuro debido a la transformación de la hemoglobina en metahemoglobina. Los índices de sedimentación sanguínea mostraron una clara variación, especialmente 60 minutos después de inuectado el veneno. Ambos fenómenos pueden estar ligados al poder lítico de los venenos. Un interesante fenómeno fue el que los animales presentaron inicialmente leucopenia, que fue seguida por persistente leucocitosis. Los presentes reultados nos llevan a concluir que el veneno de cascabel tiene un relativo poder hemolítico que se incrementa con la concentración del veneno y con la concentración de las fracciones en veneno completo