We characterized the 3272-26A-->G mutation in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, creating an alternative acceptor splice site in intron 17a, that competes with the normal one, although we predict from consensus values, with lower efficiency. We analyzed five Cystic Fibrosis (CF) Portuguese patients with the 3272-26A-->G/F508del genotype. Besides clinical and haplotype characterization of those patients, we report here results from CFTR transcript analysis in nasal brushings from all five patients. RT-PCR analysis supports alternative splicing in all patients and carriers, but not in controls. By sequencing, we determined that the alternative transcript includes 25 nucleotides from intron 17a, which predictively cause frameshift and a premature stop codon. The use of this alternative splice site causes a reduction in the levels of normal transcripts from the allele with this mutation and, most probably, of normal protein as well. By immunocytochemistry of both epithelial primary cell cultures and slices from CF polyps, CFTR protein is detected at the cell membrane, with three different antibodies. Ussing chamber analysis of one nasal polyp shows a high sodium absorption, characteristic of CF. Altogether, the results suggest that the main defect caused by the 3272-26A-->G mutation is a reduction in normal CFTR transcripts and protein and therefore this mutation should be included in class V, according to Zielenski and Tsui.

The aim of this study was to evaluate the effects of silver nitrate in the in vitro development and conservation of passion fruit plants. Experiments were carried out at the Laboratory of Culture and Tissues, in the National Cassava and Fruits Research Center (EMBRAPA), using microcuttings of Passiflora gibertii N. E. Br., previously cultivated in vitro. The microcuttings were placed in MS media supplemented with silver nitrate at concentrations of 0, 1, 2, 4 and 8 mg·L −1 , and, during 30, 60 and 90 days, the following variables were identified: Shooting length (cm), number of roots, number and coloration of leaves. A completely randomized design with 30 replications in a split-plot scheme was employed. Polynomial regression equations were adjusted in analysis of variance to compare averages of treatments. The obtained results support the assumption of high sensitivity of passion fruit tissues to ethylene, which reflects in the lost of vigor and in the induction of culture senescence by its accumulation. Based on obtained results, and in order to mitigate this problem, it is possible to indicate an addition of 2 mg·L −1 of silver nitrate in culture media, for the micropropagation of passion fruit plant, allowing the controlling of the atmosphere inside the culture test tubes, during the establishment of explants and other steps of the process.How to cite this paper: Faria, G.A., Felizardo, L

Passiflora foetida, uma espécie de maracujá silvestre, tem sido pesquisado devido ao seu potencial farmacêutico e sua característica agronômica de resistência ao Fusarium spp., possibilitando a sua utilização como porta-enxerto para espécies de maracujá suscetíveis a morte prematura. Assim, visando a propagação dessa espécie, a micropropagação é uma das formas mais utilizadas, sendo necessário a utilização de acessos de uma planta matriz para, por organogênese, gerar novas plântulas com o mesmo genótipo. Desta forma, este trabalho teve como objetivo estabelecer protocolo para a propagação in vitro de P. foetida pela indução de gemas por organogênese, testando-se duas concentrações de sais minerais do meio MS, sem adição de fitormônios.  Para o estabelecimento in vitro foram utilizados gemas apicais e laterais obtidas a partir de plantas matrizes de P. foetida, cultivadas em casa de vegetação. Os experimentos foram realizados em delineamento inteiramente casualizado. Aos 60 dias após a inoculação dos explantes foram avaliados: número de gemas, comprimento da brotação, número de folhas, escore de coloração, escore de desenvolvimento e número de raiz. Para garantir a precisão dos resultados foram realizadas técnicas estatísticas aplicadas à análise de dados e otimização dos resultados. O desenvolvimento das plantas por organogênese apresentou valores estatisticamente similares em ambas as concentrações, podendo concluir que o meio MS com metade das concentrações de nutrientes foi o mais viável para o estabelecimento in vitro dessa espécie.

A previsão do comportamento de fenômenos extremos torna-se de suma importância para toda população, devido às consequências causadas pelo aquecimento global, o que torna o estudo da Teoria de Valores Extremos (TVE) cada dia mais imprescindível em estudos de variáveis ambientais. Aplicada com a finalidade de descrever o comportamento de eventos raros, tem sido aplicada em meteorologia nas precipitações máximas, temperaturas mínimas, ventos máximos, haja vista que informações do tipo são de grande importância para o planejamento urbano, industrial, agronômico e políticas públicas mitigadoras de impacto e de desenvolvimento das cidades e da sociedade. Tendo em vista a importância do conhecimento das possíveis precipitações mensais máximas, este trabalho tem como objetivo estimar a probabilidade de precipitação pluvial mensal máxima esperada para diferentes períodos utilizando-se uma série histórica de dados pluviométricos da região de Ilha Solteira, bem como verificar o nível de ajuste dos dados ao modelo Generalized Extreme Values (GEV) utilizando o método de máxima verossimilhança para estimação dos parâmetros do modelo e verificação do ajuste em todos os períodos de chuva considerados, respectivamente. A distribuição de Gumbel foi a mais adequada para esse estudo de caso, exceto no período de estiagem da região.

The Generalized Additive Models for Location, Scale, and Shape (GAMLSS) are a recent class of models that further flexibilitythe distribution of the response variable. The regression analysis has been used to model biological phenomena, and its variousmodalities have met the need for its use with precision. However, there are situations in which the adjustment of models with moreflexible assumptions in the specification of the distribution of the response variable becomes indispensable, thus justifying the useof GAMLSS. The study of plant growth curves has full application in agricultural research; thus, it is crucial to know the habits ofgrowth and development of forest species is crucial for reforestation programs and in the most diverse researches. The study aimedto model the growth of Eucalyptus through the adjusting of Generalized Additive Models for Location, Scale, and Shape, in orderto promote improvements on crop productivity. Considering all parameters of the independent variable (time) under GAMLSSclass modeling, the distribution model ST3 presented better results.

The objective was to determine the effect of essential oils on the mortality and repellency of the Tenuipalpus heveae Baker mite (Acari, Tenuipalpidae). Leaf discs of rubber trees were deposited in Petri dishes containing a moistened cotton layer. Posteriorly, the essential oils were diluted to formulate solutions, which were sprayed onto the discs. The experimental design was completely randomized, in a 3 × 4 factorial design: neem, cedar and clove oils at doses 0, 10 μL, 50 μL and 100 μL, with five replicates, with ten adult females being considered per replicate. The exposure to the effect of essential oil was evaluated after 24, 48, 72 and 96 hours for the number of females dead and females in the cotton. The data were submitted to analysis of variance, the values were transformed into (x + 0.5)1/2, and the means were grouped by the Scott-Knot test at 5% probability for the treatments, the doses were submitted to regression analysis. From the 48 hours of evaluation, clove oil showed greater potential for adult mite mortality, being 1.38 in 48 hours, 1.40 in 72 hours and 1.50 in 96 hours. The dose of 100 μL presented a greater potential for the neem and clove oils. The clove essential oil has potential in the control of Tenuipalpus heveae.

The determination of the plot size is a practical matter pertinent to the experimental planning, and its optimal characterization allows to obtain higher precision and better quality in the results. Therefore, in this study, the main goal was to determine the plot size in experiments of passion fruit in two uniformity tests with Passiflora setacea and Passiflora alata. The experiment was constituted of a substrate at planting with 3 thirds of soil and 1 of barnyard manure. The soil was fertilizer with 3 kg of simple superphosphate and 0.5 kg of KCl by 1m³. Each species of Passiflora was considered a uniformity test with 40 basic units (BU). The evaluations of the experiments were done on 60 days after the transplant, noticing the tree's height, stem's diameter, number of leaves, number of buds, number of meristems and chlorophyll. Several plot sizes were simulated, in which each plant was first considered as a basic unit up to 40 plants per unit basic. For the estimation of optimum plot size, the maximum modified curvature method was used. The plot sizes varied with the specie, founding values as three to seven BU for Passiflora setacea and four BU to five for Passiflora alata.

An adequate experimental planning depends on several factors, such as homogeneity of the area and experimental unity, number of repetition and plot size. The adoption of an ideal plot size reduces the experimental error and, consequently, maximizes the information obtained in an experiment. Thus, the aim of the present study was to calculate the optimal plot size of eucalyptus culture in a greenhouse, in order to optimize labor and costs in the cultivation process of this economically important culture. To calculate the optimal plot size, several simulations were performed using the modified maximum curvature method. Throughout the experimental period, clonal seedlings of the Eurograndis hybrid were used and evaluated for height (H), stem diameter at the base (DB), chlorophyll content in old leaves (Chlor V) and new leaves (Chlor N) at 30, 60, 90 and 120 days after seedling transplantation. After 120 days, the seedlings were collected to determine the dry mass of the aerial part (DMAP) and the root system (DMRS). Results indicated that the optimal recommended plot size for eucalyptus, in a greenhouse, were 6 plants per plot.