Molecular characterisation of newly identified HIV-1 infections in Curitiba, Brazil: preponderance of clade C among males with recent infections

As in many areas of Brazil, the AIDS epidemic in Curitiba is relatively stable, but surveillance is important to support public policy. The molecular characteristics of HIV may be instrumental for monitoring epidemic trends. We evaluated plasma HIV-1 RNA (n = 37) from 38 cases presenting with positive serology, who were among 820 consenting volunteers visiting the downtown counselling and serology testing centre. Seroprevalence was 4.6% (CI 95% 3.2-6.3) and the estimated HIV incidence, as defined by the BED assay, was 2.86 persons/years (CI 95% 1.04-4.68). An additional set of contemporaneous, anonymous samples from a local laboratory was also analysed (n = 20). Regions of the HIV-1 polymerase (n = 57) and envelope (n = 34) were evaluated for subtyping, determination of mosaic structure, primary drug resistance mutations (pDRM), envelope V3 loop motifs and amino acid signatures related to viral tropism. HIV-1 clade B was observed in 53% of cases; HIV-1C in 30% and BC mosaics in 14%, with one F genome and one CF mosaic. Clade C infection was associated with recent infections among males (p < 0.03). Stanford surveillance pDRM was observed in 8.8% of sequences, with 7% showing high level resistance to at least one antiretroviral drug. Tropism for CXCR4 co-receptor was predicted in 18% of envelope sequences, which were exclusively among clade B genomes and cases with serological reactivity to chronic infection.

Molecular characterization of Echinococcus granulosusfrom Peru by sequencing of the mitochondrial cytochrome C oxidase subunit 1 gene

Echinococcus granulosus, the etiologic agent of cystic echinococcosis (CE) in humans and other animal species, is distributed worldwide. Ten intra-specific variants, or genotypes (G1-G10), have been defined based on genetic diversity. To determine the genotypes present in endemic areas of Peru, samples were collected from cattle (44), sheep (41) and humans (14) from Junín, Puno Huancavelica, Cusco, Arequipa and Ayacucho. DNA was extracted from protoscolex and/or germinal layers derived from 99 E. granulosus isolates and used as templates to amplify the mitochondrial cytochrome C oxidase subunit 1 gene. The resulting polymerase chain reaction products were sequenced and further examined by sequence analysis. All isolates, independent of the host, exhibited the G1 genotype. Phylogenetic analysis showed that three isolates from Ayacucho shared the same cluster with microvariant G1(4). The G1 genotype is considered the most widespread and infectious form of E. granulosusworldwide and our results confirm that the same patterns apply to this country. Therefore, these findings should be taken into consideration in developing prevention strategies and control programs for CE in Peru.

Response to treatment in Brazilian patients with chronic hepatitis C is associated with a single-nucleotide polymorphism near the interleukin-28B gene

A single-nucleotide polymorphism (SNP) upstream of interleukin (IL)28B was recently identified as an important predictor of the outcome of chronic hepatitis C patients treated with pegylated interferon plus ribavirin (PEG-IFN/RBV). The aim of this study was to investigate the association between the IL28B gene polymorphism (rs12979860) and virological response in chronic hepatitis C patients. Brazilian patients (n = 263) who were infected with hepatitis C virus (HCV) genotype 1 and were receiving PEG-IFN/RBV were genotyped. Early virological response (EVR) (12 weeks), end-of-treatment response (EOTR) (48 weeks), sustained virological response (SVR) (72 weeks) and relapse were evaluated using conventional and quantitative polymerase chain reaction (PCR) assays. The frequency of the C allele in the population was 39%. Overall, 43% of patients experienced SVR. The IL28B CC genotype was significantly associated with higher treatment response rates and a lower relapse rate compared to the other genotypes [84% vs. 58% EVR, 92% vs. 63% EOTR, 76% vs. 38% SVR and 17% vs. 40% relapse rate in CC vs. other genotypes (CT and TT), respectively]. Thus, the IL28B genotype appears to be a strong predictor of SVR following PEG-IFN/RBV therapy in treatment-naïve Brazilian patients infected with HCV genotype 1. This study, together with similar research examining other SNPs, should help to define adequate protocols for the treatment of patients infected with HCV genotype 1, especially those with a poor prognosis.

Performance of diagnostic biomarkers in predicting liver fibrosis among hepatitis C virus-infected Egyptian children

The aim of the present study was to identify specific markers that mirror liver fibrosis progression as an alternative to biopsy when biopsy is contraindicated, especially in children. After liver biopsies were performed, serum samples from 30 hepatitis C virus (HCV) paediatric patients (8-14 years) were analysed and compared with samples from 30 healthy subjects. All subjects were tested for the presence of serum anti-HCV antibodies. Direct biomarkers for liver fibrosis, including transforming growth factor-β1, tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), hyaluronic acid (HA), procollagen type III amino-terminal peptide (PIIINP) and osteopontin (OPN), were measured. The indirect biomarkers aspartate and alanine aminotransferases, albumin and bilirubin were also tested. The results revealed a significant increase in the serum marker levels in HCV-infected children compared with the healthy group, whereas albumin levels exhibited a significant decrease. Significantly higher levels of PIIINP, TIMP-1, OPN and HA were detected in HCV-infected children with moderate to severe fibrosis compared with children with mild fibrosis (p < 0.05). The diagnostic accuracy of these direct biomarkers, represented by sensitivity, specificity and positive predictive value, emphasises the utility of PIIINP, TIMP-1, OPN and HA as indicators of liver fibrosis among HCV-infected children.

Desenvolvimento e exigências térmicas de Orius insidiosus (Say) (Hemiptera, Anthocoridae)

A temperatura exerce grande influência no desenvolvimento dos insetos e o conhecimento desse aspecto é essencial para subsidiar o uso de inimigos naturais como agentes de controle biológico, bem como para a sua criação massal. O presente trabalho teve como objetivo avaliar o efeito de diferentes temperaturas no desenvolvimento de Orius insidiosus (Say, 1832), bem como as suas exigências térmicas. O experimento foi conduzido em câmaras climáticas, a 16, 19, 22, 25, 28 e 31±1&deg;C; UR de 70±10% e fotofase de 12 horas. Como alimento foram utilizados ovos de Anagasta kuehniella (Zeller, 1879). O período embrionário foi de 14,0; 8,9; 6,6; 4,8; 3,9 e 3,3 dias nas temperaturas de 16, 19, 22, 25, 28 e 31&deg;C, respectivamente. Ninfas de todos os instares (independente do sexo que deram origem) foram influenciadas pela temperatura quanto ao seu desenvolvimento, com redução nesse período com o aumento da temperatura. Machos e fêmeas, na temperatura de 25&deg;C, apresentaram um período de desenvolvimento em torno de 12 dias. A temperatura base da fase de ovo foi de 11,78&deg;C e a da fase ninfal foi de 12,27&deg;C e de 13,03&deg;C, para machos e fêmeas, respectivamente. A constante térmica para a fase de ovo foi de 63,75 e para a fase de ninfa de 161,97 e 157,24 graus-dia, para machos e fêmeas, respectivamente. A temperatura de 25&deg;C foi a mais adequada para o desenvolvimento de O. insidiosus.

Velocidade da água e a distribuição de larvas e pupas de Chirostilbia pertinax (Kollar) (Diptera, Simuliidae) e macroinvertebrados associados

O presente trabalho objetivou investigar a relação entre a velocidade da água e a distribuição dos estágios imaturos de Chirostilbia pertinax (Kollar, 1832) e os macroinvertebrados bentônicos associados. As coletas foram realizadas em cinco pontos, no trecho superior do Rio dos Sinos, no Estado do Rio Grande do Sul. A fauna bentônica foi amostrada com substratos artificiais, que permaneceram instalados nos locais de maior corenteza d'água por períodos de 14 dias. Antes do recolhimento dos substratos foram realizadas medidas de velocidade d'água no local da sua instalação. Essas medidas foram agrupadas em três classes: baixa (0,20 a 0,63 m/s), média (0,64 a 1,07 m/s) e alta (1,08 a 1,50 m/s) para posterior relação com a distribuição da fauna coletada. Para avaliar a influência da velocidade de água sobre a distribuição dos macroinvertebrados e a abundância de C. pertinax foi realizada uma regressão múltipla. As coletas resultaram em 39.598 indivíduos, sendo 33.418 simulídeos e 6.180 macroinvertebrados associados. Entre os simulídeos 5.704 espécimes correspondem a larvas do último ínstar e pupas, dos quais 828 são de C. pertinax. Quanto à distribuição dessa espécie, 30,8% ocorreram na classe de baixa velocidade d'água, 24,9% na velocidade média e 44,3% na alta velocidade. Foi observado que a abundância de C. petinax aumenta à medida que os macroinvertebrados predadores diminuem e os organismos não predadores aumentam (GLM, R² = 0,207; F 2,50 = 6,536; P = 0,003). Esses resultados revelam que os imaturos de C. pertinax têm uma tendência em ocupar um microhábitat com menor ocorrência de predadores.

Long-term C-CO2 emissions and carbon crop residue mineralization in an oxisol under different tillage and crop rotation systems

Soil C-CO2 emissions are sensitive indicators of management system impacts on soil organic matter (SOM). The main soil C-CO2 sources at the soil-plant interface are the decomposition of crop residues, SOM turnover, and respiration of roots and soil biota. The objectives of this study were to evaluate the impacts of tillage and cropping systems on long-term soil C-CO2 emissions and their relationship with carbon (C) mineralization of crop residues. A long-term experiment was conducted in a Red Oxisol in Cruz Alta, RS, Brazil, with subtropical climate Cfa (Köppen classification), mean annual precipitation of 1,774 mm and mean annual temperature of 19.2 ºC. Treatments consisted of two tillage systems: (a) conventional tillage (CT) and (b) no tillage (NT) in combination with three cropping systems: (a) R0- monoculture system (soybean/wheat), (b) R1- winter crop rotation (soybean/wheat/soybean/black oat), and (c) R2- intensive crop rotation (soybean/ black oat/soybean/black oat + common vetch/maize/oilseed radish/wheat). The soil C-CO2 efflux was measured every 14 days for two years (48 measurements), by trapping the CO2 in an alkaline solution. The soil gravimetric moisture in the 0-0.05 m layer was determined concomitantly with the C-CO2 efflux measurements. The crop residue C mineralization was evaluated with the mesh-bag method, with sampling 14, 28, 56, 84, 112, and 140 days after the beginning of the evaluation period for C measurements. Four C conservation indexes were used to assess the relation between C-CO2 efflux and soil C stock and its compartments. The crop residue C mineralization fit an exponential model in time. For black oat, wheat and maize residues, C mineralization was higher in CT than NT, while for soybean it was similar. Soil moisture was higher in NT than CT, mainly in the second year of evaluation. There was no difference in tillage systems for annual average C-CO2 emissions, but in some individual evaluations, differences between tillage systems were noticed for C-CO2 evolution. Soil C-CO2 effluxes followed a bi-modal pattern, with peaks in October/November and February/March. The highest emission was recorded in the summer and the lowest in the winter. The C-CO2 effluxes were weakly correlated to air temperature and not correlated to soil moisture. Based on the soil C conservation indexes investigated, NT associated to intensive crop rotation was more C conserving than CT with monoculture.

Efecto de la fertilización fosforada sobre el contenido de cadmio en cuatro suelos de Chile

Los fertilizantes fosforados pueden contener cadmio (Cd) y al utilizarlos pueden entrar en las cadenas tróficas. El objetivo de este estudio fue evaluar el efecto de la fertilización fosforada sobre la concentración de Cd disponible en cuatro tipos de suelos de Chile, que pertenecían a los ordenes Inceptisol, Alfisol, Ultisol y Andisol. Los dos tratamientos usados fueron el testigo sin fertilización y la aplicación de una dosis de corrección con un fertilizante comercial, superfosfato triple (SFT), que tenía una concentración de 53,2 mg de Cd por kg de fertilizante. La dosis de fertilizante aplicada fue la necesaria para alcanzar un nivel de 30 mg kg-1 de POlsen. Los suelos se incubaron en estufa durante 90 días, a 25ºC y humedad de capacidad de campo. El diseño experimental fue completamente al azar, con tres repeticiones para cada suelo y donde la unidad experimental fue un contenedor con 250 g de suelo seco. Se midió Cd disponible a los 1, 2, 7, 14, 21, 36, 49, 63, 77 y 90 días después de la incubación. La aplicación de P en dosis agronómica, utilizando fertilizante con alto contenido de Cd, tuvo un efecto estadísticamente significativo sobre la concentración de Cd disponible en los suelos estudiados, sin embargo, no hubo efecto del tiempo de incubación en la disponibilidad de Cadmio.

Secado y almacenamiento de semillas de mandarino 'Cleopatra'

Los objetivos del trabajo fueron evaluar el efecto del tratamiento con productos químicos y contenido de humedad en la conservación de semillas de mandarino 'Cleopatra' (Citrus reshni Hort ex Tan). Semillas con 14,7% de humedad tratadas con cuatro funguicidas y 8-hidroxiquinoleina, fueron almacenadas a 4±2&deg;C durante un año, al igual que tres lotes de semillas con 5,0%, 14,7% y 26,8% de humedad, tratadas con y sin 8-hidroxiquinoleina. Se evaluó la germinación y emergencia al inicio del experimento, a los seis y 12 meses de almacenamiento. En la prueba con productos químicos, el almacenamiento de semillas con 14,7% de humedad sólo fue viable hasta los seis meses, con mayores porcentajes de germinación y emergencia (59,3% y 54,0%, respectivamente), mientras que, en las semillas sin tratamiento químico fue de 33,3% y 27,3%, respectivamente. En relación con el contenido de humedad de las semillas, a los seis meses de almacenamiento se obtuvo mayor emergencia de plántulas (63,6% y 58,0%) en semillas con 26,8% y 5%, mientras que a los 12 meses, las semillas con 5%, de humedad tuvieron 75,6% de emergencia.

Diversidade genética de aceroleiras (Malphigia emarginata D.C.), utilizando marcadores moleculares RAPD e características morfoagronômicas

O conhecimento da variabilidade genética e da relação entre diferentes acessos de aceroleira é importante para maximizar o uso dos recursos genéticos para futuros programas de melhoramento. O objetivo deste trabalho foi avaliar a divergência genética entre 48 acessos de aceroleira, por meio de marcadores moleculares RAPD e características morfoagronômicas. Foram utilizados 25 iniciadores, possibilitando obter 108 marcadores, sendo observadas 92 marcas polimórficas. Os marcadores obtidos foram analisados, usando os métodos de otimização de Tocher e hierárquico UPGMA, que gerou um dendrograma utilizando o índice de Jaccard. Os resultados mostraram uma concordância parcial entre os métodos de agrupamentos estudados, com a formação de 14 grupos. Os acessos ACE 023 e ACE 033 foram os mais distintos, apresentando distância genética de 0,58. A análise comparativa dos agrupamentos revelou que os marcadores RAPD, associados com características morfoagronômicas, foram eficientes para a discriminação dos acessos e que houve uma variabilidade genética potencial para o programa de melhoramento genético e informações úteis, como a indicação de acessos promissores para avaliação clonal.

Aspectos biológicos e técnica de criação do gorgulho-da-goiaba, Conotrachelus psidii marshall (Coleoptera: Curculionidae)

Considerada praga-chave da cultura da goiaba, Conotrachelus psidii Marshall (Coleoptera: Curculionidae) tem causado grandes prejuízos em vários países produtores desta fruta. A pesquisa objetivou avaliar os parâmetros biológicos da praga, sob condições controladas, através do desenvolvimento de uma técnica de criação que permitiu efetuar observações diárias de pré-pupas e pupas. Constatou-se que o substrato adequado para manutenção dessas fases foi o solo argiloso umedecido com viabilidade de 78%, sendo este o tipo de solo utilizado na técnica de criação, que foi desenvolvida para trabalhos em câmara climatizada, à temperatura constante de 25 ± 1º C, fotoperíodo de 12 horas e UR de 70% ± 10%. Quanto aos aspectos biológicos, a duração média do período de ovo, larva, pré-pupa, pupa e ovo-pupa foi de: 4,25; 12,2; 106,6; 14,88 e 138,4 dias, respectivamente. A duração média do período da fase adulta, no solo, início da alimentação, pré-oviposição, longevidade e pós-oviposição foi de, 22; 11; 51,9; 149,32 e 7,18 dias, respectivamente. O período médio de oviposição foi de 120,27 dias, com um número médio de 343,63 ovos por fêmea. A técnica de criação desenvolvida permitiu acompanhar diariamente o desenvolvimento de pré-pupas, pupas e adultos do gorgulho-da-goiaba, sem que eles necessitassem reconstruir nova câmara, mostrando ser menos estressante e onerosa do ponto de vista energético.

PREVENTION OF CHILLING INJURY IN 'TOMMY ATKINS' MANGOES PREVIOUSLY STORED AT 5 ºC, USING HEAT TREATMENT AND RADIATION UV (UV-C)

ABSTRACT The objective of this study was to evaluate the effect of heat treatment and ultraviolet radiation (UV-C) in the prevention of chilling injury in mangoes cv. Tommy Atkins previously stored or not under injury condition after their transference to ambient condition. Fruits were divided into groups: two were hydrothermally treated (46.1 ºC/90 min; 55 ºC/5 min) and two were exposed to UV-C radiation (1.14 kJ m-2; 2.28 kJ m-2). These groups were stored under chilling injury conditions (5 ºC for 14 days), as established in preliminary tests. Other untreated groups were stored at 12 ºC or 5 ºC. After the storage period, they were transferred to ambient conditions (21.9 ºC; 55% RH) and the quality was evaluated. All the data were submitted to multivariate analysis as the tool to verify the simultaneous effect of the treatments under the quality parameters. The multivariate analysis indicated that the hydrothermal treatments at 46.1 °C/90 min and 55 °C/5 min and the UV-C radiation at doses of 1.14 kJ m-2 and 2.28 kJ m-2 were effective in minimized the symptoms of chilling injury in mangoes ‘Tommy Atkins’ stored at 5 °C for 14 days. However, after their transference to environmental condition at 21.9 °C, only the UV-C kept this control, especially at a dose of 2.28 kJ m-2. This treatment did not prevent the development of the characteristic color or affected the normal ripening and allowed the conservation of fruit for a period of 14 days at 5 °C, plus seven days of storage at environmental condition, which corresponds to the shipping transportation plus the time for sale.

Transformações químicas do (+)-10b,14-diol-allo-aromadendrano, isolado de duguetia glabriuscula r. e. fries (r. e. fries) (annonaceae) e avaliações biológicas de alguns derivados obtidos

The sesquiterpene (+)-allo-aromadendrane-10b-14-diol 1 was the lead compound to the preparation of several derivatives in order to test their biological activity against A. salina, C. sphaerospermum, E. coli and S. aureus. In this way the monoalcohols (+)-viridiflorol 4, 9 and 11 were synthesized from 1 together with the acetal 6, the ketal 7, and the ketone 8. The oxirane 3 and nitrile 5 were also prepared using as an intermediate the tosylate derivative 2.

In vitro trypanocidal evaluation of pinane derivatives from essential oils of ripe fruits from Schinus terebinthifolius Raddi (Anacardiaceae)

Essential oils of ripe fruits from Schinus terebinthifolius (Anacardiaceae), obtained using a pilot extractor and a Clevenger apparatus were chemically characterized. Due the high amount of (-)- α-pinene in both oils, this monoterpene was tested against the protozoan parasite Trypanosoma cruzi, showing a moderate potential (IC50 63.56 µg/mL) when compared to benznidazole (IC50 43.14 µg/mL). Otherwise, (-)- α-pinene oxide did not showed anti-trypanosomal activity (IC50 > 400 µg/mL) while (-)-pinane showed an IC50 of 56.50 µg/mL. The obtained results indicated that the epoxydation of α-pinene results to the loss of the anti-parasitic activity while its hydrogenation product, contributed slightly to the increased activity.

Biochemical properties of Bacillus sp. ITP-001 lipase immobilized with a sol gel process

This work presents biochemical characterization of a lipase from a new strain of Bacillus sp. ITP-001, immobilized using a sol gel process (IB). The results from the biochemical characterization of IB showed increased activity for hydrolysis, with 526.63 U g-1 at pH 5.0 and 80 ºC, and thermal stability at 37 ºC. Enzymatic activity was stimulated by ions such as EDTA, Fe+3, Mn+2, Zn+2, and Ca+2, and in various organic solvents. Kinetic parameters obtained for the IB were Km = 14.62 mM, and Vmax = 0.102 mM min-1 g-1. The results of biochemical characterization revealed the improved catalytic properties of IB.