UMEB - Unidade Móvel para Ensaio da Barra de Tração

Um equipamento denominado "Unidade Móvel de Ensaio da Barra de Tração - UMEB", foi desenvolvido na FCA/UNESP de Botucatu para realizar ensaios de tratores em solo agrícola. Construída a partir de um reboque ("trailer"), a UMEB foi adaptada para servir como carro dinamométrico instrumentado, utilizado na avaliação do desempenho de tratores submetidos a ensaios na barra de tração. Sua massa total é de 10.500 kg sustentados por um conjunto de seis rodados pneumáticos. Ensaios de campo mostraram que a UMEB proporcionou força de tração acima de 35 kN, mantendo-a constante, em diferentes condições de superfície do solo, mesmo quando a velocidade de deslocamento foi modificada.

Avaliação da resistência do solo à penetração em diferentes solos com a variação do teor de água

Com os crescimentos da população e da demanda por alimentos, novas tecnologias têm surgido visando ao aumento da produtividade e à redução nos custos de produção. Se por um lado essas tecnologias elevam a produção e reduzem os custos, por outro o tráfego intenso de máquinas e tratores alteram a estrutura do solo, tornando-o compactado, comprometendo, assim, a produtividade agrícola. Neste trabalho, relacionou-se o teor de água do solo com a sua resistência à penetração, para quatro tipos de solo e quatro condições de umidade, utilizando penetrômetro hidráulico-eletrônico com sistema de navegação DGPS. Os resultados mostraram que ocorreram modificações nas condições de atrito entre o cone e o solo com o aumento do teor de água, facilitando a penetração da haste, tornando o solo mais plástico devido à ação lubrificante entre as suas partículas. Os maiores valores de resistência do solo à penetração foram detectados mais próximos do limite de contração do solo. Houve correlação negativa entre o índice de cone e o teor de água, sendo a função de melhor ajuste a linear decrescente para todos os solos, com altos coeficientes de determinação. Nas condições de solo mais úmido, os valores de resistência do solo à penetração podem ser considerados não impeditivos para o crescimento de raízes. O tipo de solo também se mostrou significativamente influente nos resultados de índice de cone.

Agregação de ume nitossolo vermelho distroférrico sob sistemas de plantio direto, preparo convencional e mata nativa

O presente estudo teve como objetivo avaliar o efeito do tempo de adoção do sistema plantio direto, comparativamente com área de mata nativa e preparo convencional, utilizando a distribuição de agregados do solo em um Nitossolo Vermelho distroférrico. Os tratamentos foram: mata nativa (MN), preparo convencional (PC), plantio direto com um ano (PD1), plantio direto com quatro anos (PD4), plantio direto com cinco anos (PD5) e plantio direto com 12 anos (PD12). Amostras de agregados foram coletadas aleatoriamente dentro de cada tratamento, nas profundidades de 0-5 e 10-15 cm. Após o peneiramento dos agregados em água, esses foram separados nas classes de agregados de > 2 mm; < 2 mm; 2-1 mm e < 1 mm. O tempo de adoção no sistema plantio direto favoreceu a agregação do solo. O diâmetro médio ponderado (DMP) dos agregados do solo e a percentagem de agregados maiores do que 2 mm foram crescentes com o tempo de adoção no sistema plantio direto, na profundidade de 0-5 cm. A MN e o PD12 apresentaram maiores DMPs na camada de 0-5 cm. O PC apresentou maior percentual de agregados menores do que 1 mm.

Desempenho de um trator agrícola equipado com pneus radiais e diagonais com três níveis de lastros líquidos

A utilização correta de pneus em tratores, tanto em relação ao seu tipo quanto à calibração de sua pressão interna, e a lastragem ideal para cada condição de carga são fatores que influem significativamente no desempenho do trator. Esta pesquisa teve como objetivo comparar o desempenho de um trator equipado com pneus radiais e com pneus diagonais, para três condições de lastragem líquida (0%, 40% e 75% de água), em três condições superficiais de um Nitossolo Vermelho distrófico (superfície firme, preparada e com cobertura vegetal de resto de milho) e em três velocidades teóricas de deslocamento (4 km h-1, 5 km h-1 e 7 km h-1), informadas no painel do trator, correspondendo às marchas B1, B2 e C1. O melhor desempenho do trator, equipado com pneu diagonal, ocorreu na condição de 75% de água nos pneus, apresentando maior velocidade de deslocamento, menor patinhagem do trator, menor consumo horário de combustível e gerando maior potência na barra de tração. Com pneus radiais, o melhor desempenho do trator ocorreu na condição de 40% de água nos pneus, proporcionando maiores velocidades de deslocamento do trator, menores patinhagens, menores consumos, horário e específico de combustível, e maiores potência e rendimento na barra de tração.

Static tests of constructive tires using hydraulic press on the soil

ABSTRACT The power consumption and load capacity of agricultural machines have grown and the effects of pressure on the soil by tires have been still little investigated. In concern with sustainable development, the relationship machine-tire-soil must be in balance to give more consistency on the best use of tires for a given load. This study aimed to evaluate four tires of two constructive types, the bias belted tires and radial tires, both with respective rim diameters of 22.5 and 26.5 inches with variables measuring the footprint, elastic deformation, sinkage and resistance to penetration. A hydraulic press with an attachment shaft for tire mounting and a box of soil in which the tire has been imposed on a load of 53.00 kN using nominal pressures recommended by the tire manufacturer. The radial construction tire with rim diameter of 26.5 inches obtained less sinkage and resistance to penetration; however, greater elastic deformation and footprint compared to other tires. The bias-belted tire with 22.5-inch rim presented the highest resistance to penetration and the lowest elastic deformation.

Multiple forms of cotyledonary b-galactosidases from Vigna unguiculata quiescent seeds

Cotyledonary b-galactosidases were isolated and partially purified from Pitiúba cowpea (Vigna unguiculata (L.) Walp.) quiescent seeds. The purification steps consisted of precipitation of the crude extract with ammonium sulphate in the range of 20-60% saturation, acid precipitation, DEAE-Sephadex ion-exchange chromatography and Lactosyl-Sepharose affinity chromatography. This purification process gave rise to three b-galactosidases-rich fractions: b-gal I, b-gal II and b-gal III, which were purified about 5, 509, and 62 fold, respectively. They reached maximal enzyme activity at different pH ranges: 3.5-4.5 for b-gal I, 3.0-3.5 for b-gal II, and 3.0-4.0 for b-gal III. Their maximal activities were reached when the temperature of the assay medium was 60° C, and preincubation of the enzymes at different temperatures has shown that they were heat-stable up to 50° C. There were no significant differences among the partially purified enzymes as far as their response to the different effectors tested, except for Mn2+ and EDTA, which affected differently b-gal I, b-gal II, and b-gal III. They were slightly affected by Mg2+, Ca2+, Zn2+, Co2+, tartarate, molybdate, glucose, and lactose, strongly inhibited by Cu2+ and galactose, and inactivated by Hg2+. These chemical and physical properties are similar to the ones found for other plant b-galactosidases. Although through this process of purification three isoforms of this enzyme were obtained, isoelectric focusing in polyacrylamide slab gel of these enzyme-proteins suggest that cotyledons of Pitiúba cowpea quiescent seeds possess four isoforms of b-galactosidases.

Immunological properties of gene vaccines delivered by different routes

Gene vaccines represent a new and promising approach to control infectious diseases, inducing a protective immune response in the appropriate host. Several routes and methods of genetic immunization have been shown to induce antibody production as well as T helper (Th) cell and cytotoxic T lymphocyte activation. However, few studies have compared the nature of the immune responses generated by different gene vaccination delivery systems. In the present study we reviewed some aspects of immunity induced by gene immunization and compared the immune responses produced by intramuscular (im) DNA injection to gene gun-mediated DNA transfer into the skin of BALB/c mice. Using a reporter gene coding for ß-galactosidase, we have demonstrated that im injection raised a predominantly Th1 response with mostly IgG2a anti-ßgal produced, while gene gun immunization induced a mixed Th1/Th2 profile with a balanced production of IgG2a and IgG1 subclasses. Distinct types of immune responses were generated by different methods of gene delivery. These findings have important implications for genetic vaccine design. Firstly, a combination between these two systems may create optimal conditions for the induction of a broad-based immune response. Alternatively, a particular gene vaccine delivery method might be used according to the immune response required for host protection. Here, we describe the characteristics of the immune response induced by gene vaccination and the properties of DNA involved in this process.

A calcineurin inhibitory protein overexpressed in Down's syndrome interacts with the product of a ubiquitously expressed transcript

The Down's syndrome candidate region 1 (DSCR1) protein, encoded by a gene located in the human chromosome 21, interacts with calcineurin and is overexpressed in Down's syndrome patients. As an approach to clarifying a putative function for this protein, in the present study we used the yeast two-hybrid system to identify DSCR1 partners. The two-hybrid system is a method that allows the identification of protein-protein interactions through reconstitution of the activity of the yeast GAL 4 transcriptional activator. The gene DSCR1 fused to the GAL 4 binding domain (BD) was used to screen a human fetal brain cDNA library cloned in fusion with the GAL 4 activation domain (AD). Three positive clones were found and sequence analysis revealed that all the plasmids coded for the ubiquitously expressed transcript (UXT). UXT, which is encoded in human Xp11, is a 157-amino acid protein present in both cytosol and nucleus of the cells. This positive interaction of DSCR1 and UXT was confirmed in vivo by mating the yeast strain AH109 (MATa)expressing AD-UXT with the strain Y187 (MATalpha) expressing BD-DSCR1, and in vitro by co-immunoprecipitation experiments. These results may help elucidate a new function for DSCR1 and its participation in Down's syndrome pathogenesis.

Role of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivity

The role of glycosphingolipids (GSLs) present in amastigote forms of Leishmania (Leishmania) amazonensis during infection of macrophages was analyzed, with particular emphasis on GSLs presenting the terminal Galpß1-3Galpa disaccharide. Macrophage invasion by L. (L.) amazonensis amastigotes was reduced by 37% when the disaccharide Galpß1-3Galp (1 mM) was added to the culture medium. The putative macrophage receptor/lectin for ß-Gal-globotriaosylceramide (Galpß1-3Galpa1-4Galpß1-4Glc pß1-1Cer) and other structurally related GSLs from L. (L.) amazonensis amastigotes were analyzed by micelles and parasite binding assay to peritoneal macrophage proteins fractionated by SDS-PAGE under nonreducing conditions. Micelles containing purified amastigote GSLs or a suspention of L. (L.) amazonensis amastigotes fixed with 2% formaldehyde were incubated with nitrocellulose membrane containing the macrophage proteins transferred by Western blotting. Binding of micelles containing purified GSLs from amastigote forms or fixed L. (L.) amazonensis amastigotes to nitrocellulose membrane was probed using monoclonal antibody ST-3, which recognizes the glycoepitope Galpß1-3Galpa1-R present either in the micelle preparation or on the amastigote surface. Macrophage protein with molecular mass ~30 kDa bound the amastigote GSL and appeared to be a doublet on electrophoresis. The specificity of this interaction was confirmed using fixed L. (L.) chagasi amastigotes, which do not express GSLs such as ß-Galp-globotriaosylceramides, and which do not bind to 30-kDa protein.

Transient high-level expression of ß-galactosidase after transfection of fibroblasts from GM1 gangliosidosis patients with plasmid DNA

GM1 gangliosidosis is an autosomal recessive disorder caused by the deficiency of lysosomal acid hydrolase ß-galactosidase (ß-Gal). It is one of the most frequent lysosomal storage disorders in Brazil, with an estimated frequency of 1:17,000. The enzyme is secreted and can be captured by deficient cells and targeted to the lysosomes. There is no effective treatment for GM1 gangliosidosis. To determine the efficiency of an expression vector for correcting the genetic defect of GM1 gangliosidosis, we tested transfer of the ß-Gal gene (Glb1) to fibroblasts in culture using liposomes. ß-Gal cDNA was cloned into the expression vectors pSCTOP and pREP9. Transfection was performed using 4 µL lipofectamine 2000 and 1.5-2.0 µg DNA. Cells (2 x 10(5)/well) were harvested 24 h, 48 h, and 7 days after transfection. Enzyme specific activity was measured in cell lysate and supernatant by fluorometric assay. Twenty-four hours after transfection, treated cells showed a higher enzyme specific activity (pREP9-ß-Gal: 621.5 ± 323.0, pSCTOP-ß-Gal: 714.5 ± 349.5, pREP9-ß-Gal + pSCTOP-ß-Gal: 1859.0 ± 182.4, and pREP9-ß-Gal + pTRACER: 979.5 ± 254.9 nmol·h-1·mg-1 protein) compared to untreated cells (18.0 ± 3.1 for cell and 32.2 ± 22.2 nmol·h-1·mg-1 protein for supernatant). However, cells maintained in culture for 7 days showed values similar to those of untreated patients. In the present study, we were able to transfect primary patients' skin fibroblasts in culture using a non-viral vector which overexpresses the ß-Gal gene for 24 h. This is the first attempt to correct fibroblasts from patients with GM1 gangliosidosis by gene therapy using a non-viral vector.

Galantamine protects against lipopolysaccharide-induced acute lung injury in rats

Lipopolysaccharide (LPS)-induced endotoxemia triggers the secretion of proinflammatory cytokines and can cause acute lung injury (ALI). The high mobility group box 1 (HMGB1) protein plays an important role as a late mediator of sepsis and ALI. Galantamine (GAL) is a central acetylcholinesterase inhibitor that inhibits the expression of HMGB1. This study evaluated the effects of GAL by measuring levels of inflammatory mediators and observing histopathological features associated with LPS-induced ALI. Sixty 8-10 week old male Sprague-Dawley rats (200-240 g) were randomized into three groups as follows: control group, LPS group (7.5 mg/kg LPS), and LPS+GAL group (5 mg/kg GAL before LPS administration). Histopathological examination of lung specimens obtained 12 h after LPS administration was performed to analyze changes in wet-to-dry (W/D) weight ratio, myeloperoxidase (MPO) activity, and HMGB1 expression level. Additionally, plasma concentrations of tumor necrosis factor-α, interleukin-6, and HMGB1 were measured using an enzyme-linked immunosorbent assay at 0 (baseline), 3, 6, 9, and 12 h after LPS administration. Mortality in the three groups was recorded at 72 h. LPS-induced ALI was characterized by distortion of pulmonary architecture and elevation of MPO activity, W/D weight ratio, and levels of pro-inflammatory cytokines, including tumor necrosis factor-α, interleukin-6, and HMGB1. Pretreatment with GAL significantly reduced the LPS-induced lung pathological changes, W/D weight ratio, levels of pro-inflammatory cytokines and MPO activity (ANOVA). Moreover, GAL treatment significantly decreased the mortality rate (ANOVA). In conclusion, we demonstrated that GAL exerted a protective effect on LPS-induced ALI in rats.

Aloe vera extract as a promising treatment for the quality maintenance of minimally-processed table grapes

The effect of an edible film obtained from a commercial Aloe vera extract, on the quality maintenance of minimally processed grapes belonging to three different cultivars (Sugar One, Victoria and Black Magic) was evaluated by enzymatic (PPO, PME, β-GAL), physicochemical (pH, acidity, °Brix), and sensorial methods. All the analyzed parameters were measured in extracts obtained from minimally processed grapes packaged in ordinary atmosphere and stored at 4 °C for 15 days. Samples dipped into Aloe vera showed significant differences (p≤0.05) compared to untreated ones. The determination of such parameters and the evaluation of consumer acceptability were helpful to determine the effectiveness of the post-harvest treatment with Aloe vera for a storage period of 15 days.

Podocitúria na doença de Fabry

Resumo Introdução: A doença de Fabry (DF) é uma desordem lisossômica ligada ao cromossomo X ocasionada por mutações no gene que codifica a enzima lisossômica α-galactosidase A (α-GAL). A redução ou ausência da atividade dessa enzima leva ao acúmulo progressivo de gb3. A doença renal é uma importante consequência clínica da acumulação de Gb3. Podócito é o tipo celular mais afetado na doença renal, que mostra apenas uma resposta parcial à Terapia de Reposição Enzimática. Além disso, a disfunção podocitária é a principal contribuinte para a perda progressiva da função renal e pode ser encontrada alterada mesmo antes do início da microalbuminúria. Assim, a podocitúria na DF pode ser uma ferramenta importante para prever a doença renal. Objetivo: O objetivo deste estudo foi quantificar a excreção urinária de podócitos em pacientes com DF (V269M, n = 14) e controles saudáveis (n = 40), e relacioná-las com as variáveis sexo, idade, tempo de terapia e a razão albumina: creatinina (AUC). Métodos: Podócitos urinários foram identificados utilizando imunofluorescência para podocalixina e DAPI. O número de células podocalixina positivo foi contado e o número médio foi utilizado (faixa normal 0-0.6 podócitos/mL). Resultados: O número médio de podócitos na urina de pacientes com DF foi significativamente maior do que os controles saudáveis (p < 0.0001). Observou-se uma correlação positiva entre podocitúria e AUC (p = 0.004; r2 = 0.6417). Conclusão: A podocitúria pode ser uma ferramenta adicional para avaliar a progressão da doença renal em pacientes que se espera que tenha um fenótipo mais agressivo.