AAV5-mediated gene transfer to the parotid glands of non-human primates

Salivary glands are potentially useful target sites for multiple clinical applications of gene transfer. Previously, we have shown that serotype 2 adeno-associated viral (AAV2) vectors lead to stable gene transfer in the parotid glands of rhesus macaques. As AAV5 vectors result in considerably greater transgene expression in murine salivary glands than do AAV2 vectors, herein we have examined the use of AAV5 vectors in macaques at two different doses (n3 per group; 1010 or 3 × 1011 particles per gland). AAV5 vector delivery, as with AAV2 vectors, led to no untoward clinical, hematological or serum chemistry responses in macaques. The extent of AAV5-mediated expression of rhesus erythropoietin (RhEpo) was dose-dependent and similar to that seen with an AAV2 vector. However, unlike results with the AAV2 vector, AAV5 vector-mediated RhEpo expression was transient. Maximal expression peaked at day 56, was reduced by & ∼80% on day 84 and thereafter remained near background levels until day 182 (end of experiment). Quantitative PCR studies of high-dose vector biodistribution at this last time point showed much lower AAV5 copy numbers in the targeted parotid gland (& ∼1.7%) than found with the same AAV2 vector dose. Molecular analysis of the conformation of vector DNA indicated a markedly lower level of concatamerization for the AAV5 vector compared with that of a similar AAV2 vector. In addition, cellular immunological studies suggest that host response differences may occur with AAV2 and AAV5 vector delivery at this mucosal site. The aggregate data indicate that results with AAV5 vectors in murine salivary glands apparently do not extend to macaque glands. © 2010 Macmillan Publishers Limited All rights reserved

Desempenho e uniformidade da distribuição de água de um pivô central Performance of a center pivot irrigation system and irrigation distribution uniformity

O objetivo deste experimento foi avaliar as características de desempenho de um equipamento de irrigação pivô central, de baixa pressão, bem como determinar a uniformidade de distribuição de água, tanto abaixo como acima da superfície do solo. Para a avaliação do desempenho do sistema utilizaram-se os coeficientes de uniformidade de Christiansen (CUC) e de distribuição (CUD), calculados a partir de dados da precipitação dos aspersores. O pivô central foi ensaiado nas velocidades de 25%, 50%, 75% e 100% da velocidade máxima de deslocamento do equipamento. Utilizou-se quatro linhas de coletores, uniformemente espaçados. Os valores encontrados do CUC e o CUD foram superiores ao mínimo recomendado para a cultura do milho, confirmando o bom desempenho do pivô central. Em todas as profundidades do solo estudadas os coeficientes de uniformidade foram superiores aos obtidos acima do solo, ocorrendo um aumento nos valores dos coeficientes de uniformidade abaixo da superfície do solo com o tempo.<br>The objective of this work was to evaluate some characteristics of a center pivot irrigation system equipament, as well as to determine the water distribuition uniformity, under and over the soil surface. The Christiansen uniformity coefficient (CUC) and distribuition (CUD) were used to evaluate the system. A low pressure center pivot was tested in four different speeds: 25%, 50%, 75% and 100% of the timer sensor, and in four collectors Unes spaced. Results indicated that both coeficients presented good performance. The values of CUC and CUD were above the minimum recommended for na irrigated corn field. The uniformity coefficients in all depths of the soil were higher than the above the soil surface. Howerer, the uniformity under soil surface increased with the time in all depths

Diversidade genética de Begomovirus que infectam plantas invasoras na região nordeste Genetic diversity of Begomovirus infecting weeds in northeastern Brazil

Os Begomovirus fazem parte de uma família numerosa de fitovírus denominada Geminiviridae. Eles infectam ampla gama de hospedeiras, incluindo muitas espécies cultivadas, como tomate (Lycopersicon esculentum), feijão (Phaseolus vulgaris), pimentão (Capsicum annuum), caupi (Vigna unguiculata), mandioca (Manihot esculenta) etc., além de plantas invasoras de várias espécies. Em alguns casos, plantas invasoras podem funcionar como reservatórios desses vírus para plantas cultivadas, mediante transmissão pelo inseto-vetor. No presente trabalho, plantas invasoras com sintomas de mosaico amarelo, deformação do limbo foliar e redução do crescimento foram avaliadas no tocante à presença de Begomovirus mediante a técnica de PCR, empregando-se oligonucleotídeos universais para detecção desses vírus. Foram avaliadas 11 amostras, correspondendo a 10 espécies, coletadas em municípios dos Estados de Alagoas, Pernambuco e Bahia. Algumas, como Herissantia crispa, Waltheria indica e Triumfetta semitriloba, são relatadas pela primeira vez como espécies hospedeiras de Begomovirus. Para estimar a variabilidade genética dos Begomovirus detectados, o produto de amplificação dos diversos isolados foi clivado com as enzimas de restrição EcoRI, HinfI e TaqI. Confirmando resultados obtidos para plantas cultivadas por outros grupos de pesquisa, foram observados padrões distintos de clivagem para os isolados estudados, evidenciando a grande variabilidade genética desses vírus.<br>Genus Begomovirus belong to the family Geminiviridae. Begomovirus is associated with a wide range of hosts, including many cultivated species such as tomato (Lycopersicon esculentum), dry beans (Phaseolus vulgaris), pepper (Capsicum annuum), cowpea (Vigna unguiculata), cassava (Manihot esculenta), etc., besides many weed species. It has been demonstrated that in some cases weeds act as virus reservoirs for cultivated plants. In the present work, weed samples presenting yellow mosaic, foliar malformation and size reduction were tested by PCR for infection by Begomovirus, using specific degenerate oligonucleotides. Eleven samples corresponding to 10 plant species were collected in the countryside towns in the states of Alagoas, Pernambuco and Bahia. Some plant species such as Herissantia crispa, Waltheria indica and Triumfetta semitriloba are reported for the first time as hosts for Begomovirus. To estimate the genetic diversity of the detected Begomovirus, the amplified products of several isolates were cleaved with each three restriction enzymes, EcoRI, HinfI, and TaqI. Different patterns were observed for the studied isolates, pointing out to a great genetic diversity for these viruses