Genetic diversity of NS3 protease from Brazilian HCV isolates and possible implications for therapy with direct-acting antiviral drugs

The hepatitis C virus (HCV) NS3 protease has been one of the molecular targets of new therapeutic approaches. Its genomic sequence variability in Brazilian HCV isolates is poorly documented. To obtain more information on the magnitude of its genetic diversity, 114 Brazilian HCV samples were sequenced and analysed together with global reference sequences. Genetic distance (d) analyses revealed that subtype 1b had a higher degree of heterogeneity (d = 0.098) than subtypes 1a (d = 0.060) and 3a (d = 0.062). Brazilian isolates of subtype 1b were distributed in the phylogenetic tree among sequences from other countries, whereas most subtype 1a and 3a sequences clustered into a single branch. Additional characterisation of subtype 1a in clades 1 and 2 revealed that all but two Brazilian subtype 1a sequences formed a distinct and strongly supported (approximate likelihood-ratio test = 93) group of sequences inside clade 1. Moreover, this subcluster inside clade 1 presented an unusual phenotypic characteristic in relation to the presence of resistance mutations for macrocyclic inhibitors. In particular, the mutation Q80K was found in the majority of clade 1 sequences, but not in the Brazilian isolates. These data demonstrate that Brazilian HCV subtypes display a distinct pattern of genetic diversity and reinforce the importance of sequence information in future therapeutic approaches.

Relationship between grain colour and preharvest sprouting-resistance in wheat

Since red alleles (R) of the genes that control grain colour are important for the improvement of preharvest sprouting resistance in wheat and there are three independently inherited loci, on chromosomes 3A, 3B and 3D of hexaploid wheat, it is possible to vary the dosage of dominant alleles in a breeding program. The objective of this work was to evaluate the dosage effect of R genes on preharvest sprouting, in a single seed descent population, named TRL, derived from the cross between Timgalen, white-grained wheat, and RL 4137, red-grained wheat. The study was carried out using sprouting data in ripe ears obtained under artificial conditions in a rainfall simulator over three years. According to the results there is a significant effect on preharvest sprouting provided by colour and a weaker effect of increasing R dosage. However, the significant residual genotypic variation between red lines and all lines (reds and whites) at 0.1% level showed that preharvest sprouting was also controlled by other genes. There are no significant correlations between sprouting and date of ripeness or between ripeness, R dosage and colour intensity.

Características agronômicas de seis genótipos de maracujazeiro-azedo cultivados no Distrito Federal

Este trabalho teve como objetivo avaliar as características agronômicas de seis genótipos de maracujazeiro-azedo cultivados no Distrito Federal. O experimento foi desenvolvido na área experimental da Fazenda Água Limpa da Universidade de Brasília, no DF. Utilizou-se o delineamento em blocos casualizados, com quatro repetições, seis tratamentos e sete plantas por parcela. Foram avaliados os genótipos: Rubi Gigante, EC-3-0, EC-L-7, RC-3, Redondão e Gigante Amarelo. Os parâmetros analisados, durante seis meses (17 colheitas), foram: produtividade total estimada, quantidade de frutos e coloração da casca. O genótipo Rubi Gigante teve a maior produtividade (16,69 t/ha-1), enquanto o RC-3 a menor (2,92 t/ha-1). Foram observados resultados semelhantes quanto às variáveis: produtividade total e quantidade de frutos de tamanho: primeira, 1B e 1A em relação aos seis genótipos. Não houve diferença significativa a 5%, pelo teste de Tukey, para: % de frutos de coloração amarela, rosa e roxa; massa fresca de frutos amarelos e rosas; quantidade de frutos rosas e roxos, e também quantidade de frutos de tamanho 2A e 3A em relação a todos os genótipos avaliados. Os genótipos Rubi Gigante, Redondão, EC-3-0 e EC-L-7 podem ser recomendados para o cultivo no DF e região geoeconômica, se confirmarem a superioridade quando avaliados por um maior período e em diferentes condições ambientais.

Síntesis de espirobrasinoesteroides análogos de la 6-desoxocastasterona

The synthesis of two new spirostanic analogs of the natural occurring brassinosteroid 6-desoxocastasterone (1) is described. The scheme consists in the formation and elimination of tigogenin mesylate followed by catalytic dihydroxylation of the resulting D2-steroid (3) and acetylation of the 2a, 3a-diol introduced.Treatment diacetate (5) with NaNO2/BF3.Et2O and chromatography in alumina led to a 23-keto (6) which on reduction produced the 23S alcohol (8) as major product. Saponification of the 2a, 3a-diacetoxy-23-keto compound (6) and the 2a,3a-diacetoxy-23-hydroxy compound (8) led to the spirobrasinosteroids (7) and (9).13C NMR and ¹H RMN characteristics derived from substitution at C23 are briefly discussed.

Síntese e avaliação da atividade fitotóxica de derivados da α-Santonina

Mixtures of α-Santonin and various solvents were irradiated by either high or low pressure mercury lamps. The photochemical reactions afforded lumisantonin (11) (76% in acetonitrile), (3S,3aS,9bS)-3,6,6-trimethyl-3,3a,4,5-tetrahydronafto[1,2-b]furan-2,7(6H,9bH)dione (12) (100% in acetonitrile), 10α-acetoxy-3-oxo-1,7αH,6,11βH-guaia-4-en-6,12-olide (8) (26% in acetic acid), 10α-hydroxy-3-oxo-1,7αH,6,11βH-guaia-4-en-6,12-olide (10) (32%) and (E)-3-((3S,3aS,7aS)-3-methyl-2-oxo-6-(propan-2-ylidene)hexahydrobenzofuran-7-(7aH)-ylidene)propanoic acid (9) (44%) (in water/ acetic acid 1:1, v/v). Lactone 12 was also prepared by irradiation of lumisantonin in diethyl ether. Lactones 8 and 10 were converted, respectively, into the 10α-acetoxy-3β-hydroxy-1,7αH,6,11βH-guaia-4-en-6,12-olide (13) (87%) and 3β,10α-dihydroxy-1,7αH,6,11βH-guaia-4-en-6,12-olide (14) (75%) by sodium borohydride reduction. The effects of the compounds on the development of radicle of Sorghum bicolor and Cucumis sativus were evaluated.

Polarizabilidades e primeira hiperpolarizabilidades elétricas dipolares de ésteres E-4-amino-trans-1,3-butadienil-(1',2'-di-hidroxibenzeno) de boro, alumínio e gálio

Static electric dipole polarizabilities and first hyperpolarizabilites have been calculated for the title molecules and their 3' and 4'-nitro derivatives at ab-initio Hartree- Fock/6-31G(d, p) level. The influence of the pivotal p vacant 3A elements (B, Al or Ga) substitution on the electrical properties of these molecules is detailed. The axial vector components of the first hyperpolarizabilities β(0) of the push-pull 4'-nitro derivatives, -18.2×10-32 esu (B), -21.1×10-32 esu (Al) and -20.8×10-32 esu (Ga) are calculated to be as much as fourfold larger then that calculated for the p-nitroaniline, a reference organic molecule for comparison for this type of molecular property.

Synthesis, antimicrobial and cytotoxic activities of 5-benzylidene-2-[(pyridine-4-ylmethylene)hydrazono]-thiazolidin-4-one and 2-[(pyridine-4-ylmethylene) hydrazono]-thiazolidin-4-one derivatives

A new series of 5-benzylidene-2-[(pyridine-4-ylmethylene)hydrazono]-thiazolidin-4-ones 4a-l have been synthesized. These compounds were designed by a molecular hybridization approach. 2-[(Pyridine-4-ylmethylene)hydrazono]-thiazolidin-4-ones 3a-d were also obtained and used as intermediates to give the target compounds. The in vitro antimicrobial and cytotoxic activities were evaluated for both series. The intermediate 3b showed considerable antibiotic activity against B. subtilis and C. albicans. In the cytotoxic activity compounds 3b (IC50= 4.25 ± 0.36 µg/mL) and 4l (IC50= 1.38 ± 0.04 µg/mL) were effective for inhibition of human erythromyeloblastoid leukemia (K-562) and human lung carcinoma (NCI-H292) cell lines, respectively.

Estudo em raiz e ráquis foliar de spathelia excelsa: fitoquímica e atividade frente ao fungo Moniliophthora perniciosa associado ao cupuaçuzeiro (Theobroma grandiflorum)

The chemical composition of Spathelia excelsa (Krause) R. S. Cowan & Brizicky was investigated and the limonoids harrisonin (1) and deacetylspathelin (2), alkaloids folinin and casimiroin mixture (3a, b), plus a further casimiroin (3b) were identified in methanol extract from root. The CH2Cl2 extract from the rachis yielded protolimonoid 3β-angeloyl-21,24-epoxy-7α,21α,23α,25-tetrahydroxy-4α,4β,8β,10β-tetramethyl-25-dimethyl-14,18-cyclo-5α,13α,14α,17α-cholestane (4), and methanol extract, the limonoids limonin diosphenol (5) and perforatin (6), as well as the chromone biflorin (7). Harrisonin and biflorin were isolated for the first time in this genus. On the antifungal assay against witches' broom (Moniliophthoraperniciosa) compound 3b was found to be active.

Variability of the wheat powdery mildew pathogen Blumeria graminis f. sp. tritici in the 2003 crop season

Wheat (Triticum aestivum) powdery mildew, caused by the biotrophic fungus Blumeria graminis f. sp. tritici, is one of the most severe foliar diseases attacking this crop, reducing grain yields by 10% to 62% in Brazil. The disease can be controlled by genetic resistance of the host, but the pathogen has physiological specialization, which enables it to infect wheat cultivars that have remained resistant for years. The objective of this work was to evaluate the variability of pathogenic strains of B. graminis f. sp. tritici collected in Brazil and the effectiveness of wheat resistance genes to powdery mildew in the 2003 crop season. Plants of a differential series were inoculated with each monopustular isolate. Thirty-one combinations of effective and ineffective resistance genes were identified. Only the gene Pm4a+... remained totally effective to all isolates, and gene Pm6 was highly effective (below 10% of susceptibility), whereas genes Pm3a and Pm8 were totally ineffective (susceptible to all isolates). Genes Pm3c, D1, and D2 showed low effectiveness (above 50% of susceptibility), and genes Pm1, 2, 4a, 1+?, and 2+Mld had mean effective results to most strains (susceptibility between 10% and 49%). The virulence formula Pm1, 3c, 4a, 6, 1+?, 2+Mld, 4a+..., D2 (effective genes) / 2, 3a, 8, D1 (ineffective genes) was most frequently found, accounting for 15% of the occurrences. The most frequent number of ineffective genes was seven, ranging from three to ten.

Pequisas físicas e químicas do sistema hidrográfico da região lagunar de Cananéia: I - Cursos de águas

The present elaboration is a "previous note" about two watercourses, Maria Rodrigues and Baguaçú, belonging to Hydrografic System of the Cananeas Lagoon Region in the Southern Part of it he Coast of the Estado de São Paulo, Brazil, which is the objete of our physical and chemical researches. It is based on the first results obtained from our contact with that region, ttfie studies of which are being continued and will still have to confirm or not the previous conclusions, which we are presenting in this "provisional paper". We studied the Maria Rodrigues and the Baguacu under the discussion whether they are to be clasified as a river in the poper sense ot the word or waUer-courses of any other origin. For this purpose we established observatory-stations along their whole course where we collected quantities of water to be analised. The data included in the table nº 1 and 2 refer to the physical and chemical properties of these portions of water. These physical and chemical properties were discussed and compared one with the other according to the observatory-stations, at the same moment and the tide predominating on that occasion. The physical and chemical datas were obtained according to the norms recommended by the Conseil International permanent pour l'Exploration de la Mer, Copenhague. We also presented a sketch of both; we described their relations with regard to the Mar de Cananéia; we inserted comprehensible histograms of the distribution of the observatory-stations, each of them with the respective indices, and at last we tried to correlate the occurrence of the "plancton" with the physical and chemical properties of water as well in the Maria Rodrigues as in the Baguacú. From these researches result the following previous conclusions: 1 - By the data obtained up to now nothing could be formulated as to a cyclical or "seasonal" aspect. 2 - By the comparison of the phenomena which present the rivers in the proper sense of the word and those observed in the Maria Rodrigues and the Baguacú, we are convinced that both are water-courses of the classical type "marigot,". 3 - Their channels are maintained open by the effect of the tide and, more perphaps, by the infiltration-water deriving from the underground-water level owing to the type of the soil of that region, which in mean shows an elevation of 2-4 meters over sea-level. 4 - From tables 1 and 2 and histograms 2 and 3 we stated that the water, not only of the Maria Rodrigues but also of the Baguacú, conserve more or less the same physical and chemical properties, provided that: 1) the localities to be compared are the mouths or equidistant points from them in the lower course, and 2) the moment and tide must be the same. Tables 3a and 3b show sufficiently this supposition.

Pseudomonas syringae pv. tabaci in papaya seedlings

The natural occurrence of Pseudomonas syringae pv. tabaci causing leaf spot symptoms in papaya seedlings is reported. The pathogen was identified through biochemical, physiological, serological, and molecular assays and artificial inoculations in papaya plants. It was also shown that the strains were pathogenic to bean and tobacco plants. The restriction patterns obtained with Afa I, Alu I, Dde I, Hae III, Hpa II, Hinf I, Sau 3A I and Taq I of the PCR-RFLP of 16S-23S DNAr were identical to the P. s. pv. tabaci patterns. Primers corresponding to hrpL gene of P. syringae were also tested and the results grouped the papaya strains with P s. pv. tabaci. Bacterial strains were deposited at Coleção de Culturas IBSBF, Instituto Biológico, Campinas, Brazil, under access numbers 1687 and 1822.

Influence of different systems of minimum ventilation on air quality in broiler houses

To maintain a competitive development of the Brazilian aviculture, some measures must be taken to enable the identification and reduction of risks to the health of birds, as well as for the physical environment. Therefore, the aim of this study was to evaluate the effectiveness of three different systems of minimum ventilation (positive pressure - SVMP, negative pressure - SVMN and natural ventilation - SVMNat) in the air quality during the first 21 days of life of broiler chicks, during winter. Three points were selected along the length to verify ammonia, carbon monoxide and oxygen concentrations at 3a.m., 9a.m., 3p.m. and 9p.m., by the respiration level of birds and workers. The averages of pollutant gases did not exceed the tolerance levels for the three minimum ventilation systems evaluated, which is 20 and 10ppm for the birds level and 20 and 39ppm for the workers level, for ammonia and carbon monoxide, respectively. It was evident that the minimum ventilation systems were appropriately sized for the required minimum ambient air renovation, in respect to ventilation rates applied for the SVMN and SVMP systems, and with air velocity at levels that do not stress the chicks, including for the SVMNat. The three studied systems of minimum ventilation allowed the birds to externalize their productive performance, with values close to the ones considered satisfactory for all evaluated parameters, in accordance with the Brazilian aviculture standards.

Efeito da ventilação com diferentes frações inspiradas de oxigênio e do alopurinol na isquemia-reperfusão pulmonar em ratos

OBJETIVO: Avaliar o efeito da ventilação associada a frações inspiradas de oxigênio a 0,21 e 1,00 e do alopurinol (antioxidante) na isquemia-reperfusão pulmonar. MÉTODO: Foram utilizados 60 ratos Wistar, distribuídos aleatoriamente em seis grupos. O grupo 1 foi o controle; no grupo 2 os animais foram ventilados durante a isquemia-reperfusão pulmonar com FiO2 de 0,21; e no grupo 3, com FiO2 de 1,00. Os três grupos restantes 1A, 2A e 3A foram medicados com 100 mg/kg de alopurinol no pré-operatório e submetidos a procedimentos semelhantes aos grupos 1, 2 e 3, respectivamente. O modelo utilizado foi de isquemia-reperfusão normotérmica, in situ. O tempo de isquemia foi de 30 minutos, e o de reperfusão, de 10 minutos. Como parâmetros de avaliação foram utilizados a pressão arterial média sistêmica (PAM), a relação da pressão parcial de oxigênio/fração inspirada de oxigênio (PaO2/FiO2), a dosagem das substâncias reativas ao ácido tiobarbitúrico (TBARS) no tecido pulmonar e a relação entre peso pulmonar úmido e peso pulmonar seco. RESULTADOS: Em relação à PAM, ocorreu diminuição significante (p<0,05) entre os grupos 3 x 1, 2 x 2A e 3 x 3A. Na PaO2/FiO2 ocorreu diminuição significante (p<0,05) entre os grupos 3 x 2 e 3 x 3A. Nas TBARS ocorreu diminuição significante (p<0,05) entre os grupos 3 x 3A. Na relação peso pulmonar úmido/seco ocorreu aumento significante (p<0,05) entre os grupos 3 x 2, 2 x 2A e 3 x 3A. CONCLUSÕES: A ventilação com oxigênio a 21%, quando comparada à ventilação com oxigênio a 100%, apresentou diminuição menos acentuada da PAM, melhor relação entre PaO2/FiO2, e menor edema pulmonar. O uso de alopurinol no pré-operatório mostrou uma diminuição menos acentuada da PAM, melhor relação entre PaO2/FiO2, menor produção de TBARS e menor edema pulmonar, quando comparado aos resultados dos grupos que não o utilizaram.

Comparação entre as técnicas de shouldice modificada por berliner e a de falci-lichtenstein na hernioplastia inguinal

OBJETIVO: Comparar os resultados da hernioplastia inguinal pelas técnicas de Shouldice modificada por Berliner (SB), que realiza dois planos de sutura superpostos e de Falci-Lichtenstein (FL) que utiliza prótese de polipropileno. MÉTODO: Foram estudados prospectivamente 312 pacientes operados pelo mesmo cirurgião entre 1997 e 2004. O Grupo 1 constou de 84 pacientes operados pela técnica de FL e o Grupo 2 foi formado por 228 pacientes operados pela técnica de SB. Todos eram masculinos, maiores de 18 anos e portadores de hérnias inguinais tipo 3A, 3B e 4 da classificação de Nyhus. Os grupos foram semelhantes em relação a idade, lado operado, tipo de hérnia, anestesia empregada e tempo de seguimento (média de 3,5 anos). RESULTADOS: A duração média dos procedimentos foi de 53 min no Grupo 1 e de 57 min no Grupo 2 (p=0,2982); a permanência hospitalar foi menor que 24h para 94% dos pacientes do Grupo 1 e 92% do Grupo 2 (p=0,8050); a incidência de complicações foi de 9,5% no Grupo 1 e de 12,3% no Grupo 2 (p=0,5557) sendo a mais comum o hematoma/equimose e a taxa de recidiva foi de 1,2% no Grupo 1 e de 5,4% no Grupo 2 (p=0,0935). Nenhum desses resultados apresentou diferença significante. CONCLUSÃO: A técnica SB mostrou-se comparável à técnica de FL em homens, maiores de 18 anos com hérnias tipo 3 A, 3 B e 4 de Nyhus, além de não exigir material protético.

Hepatitis C virus infection in a Brazilian population with sickle-cell anemia

Patients with sickle-cell anemia submitted to frequent blood transfusions are at risk of contamination with hepatitis C virus (HCV). Determination of HCV RNA and genotype characterization are parameters that are relevant for the treatment of the viral infection. The objective of the present study was to determine the frequency of HCV infection and the positivity for HCV RNA and to identify the HCV genotype in patients with sickle-cell anemia with a history of blood transfusion who had been treated at the Hospital of the HEMOPE Foundation. Sera from 291 patients were tested for anti-HCV antibodies by ELISA 3.0 and RIBA 3.0 Chiron and for the presence of HCV RNA by RT-PCR. HCV genotyping was performed in 19 serum samples. Forty-one of 291 patients (14.1%) were anti-HCV positive by ELISA and RIBA. Both univariate and multivariate analysis showed a greater risk of anti-HCV positivity in those who had started a transfusion regime before 1992 and received more than 10 units of blood. Thirty-four of the anti-HCV-positive patients (34/41, 82.9%) were also HCV RNA positive. Univariate analysis, used to compare HCV RNA-negative and -positive patients, did not indicate a higher risk of HCV RNA positivity for any of the variables evaluated. The genotypes identified were 1b (63%), 1a (21%) and 3a (16%). A high prevalence of HCV infection was observed in our patients with sickle-cell anemia (14.1%) compared to the population in general (3%). In the literature, the frequency of HCV infection in sickle-cell anemia ranges from 2 to 30%. The serological screening for anti-HCV at blood banks after 1992 has contributed to a better control of the dissemination of HCV infection. Because of the predominance of genotype 1, these patients belong to a group requiring special treatment, with a probable indication of new therapeutic options against HCV.