Effect of environmental mycobacteria (Mycobacterium avium) on immunity induced by a DNA vaccine (DNAhsp65) against tuberculosis

The efficacy of BCG vaccine (attenuated Mycobacterium bovis) against pulmonary tuberculosis varies enormously among different populations. The prevailing hypothesis attributes this variation to interactions between the vaccine and mycobacteria common in the environment. Studies have revealed that most protective antigens expressed by the antituberculous vaccine are conserved in M. avium, supporting the hypothesis that exposure to environmental mycobacteria generates a cross-reactive immune response that interferes with BCG efficacy. In this study we investigated the effect of a prior exposure to heat-killed M. avium on the immune response and the protective efficacy induced by a genetic vaccine pVAXhsp65 (hsp65 gene from M. leprae inserted in pVAX vector) against experimental tuberculosis. To evaluate the effect on the immune response, female BALB/c mice were initially injected with distinct doses (0.08×106, 4×106, and 200×10 6) of heat-killed M. avium by subcutaneous route. Three weeks later, the animals were immunized with 3 doses of DNAhsp65 by intramuscular route (100μg/15 days apart). Control groups received only M. avium, vaccine (pVAXhsp65), vector (pVAX) or saline solution. Cytokine production and antibody levels were determined by ELISA. To evaluate the effect on the protective efficacy, animals were initially sensitized with 200×106 heat-killed CFU of M. avium by subcutaneous route and then immunized with 3 doses of pVAXhsp65 (100μg/15 days apart) by intramuscular route. Control groups were injected with saline, pVAX (4 doses), pVAXhsp65 (4 doses), M. avium or M. avium plus pVAX (3 doses). Fifteen days after last DNA dose, the animals were infected with 1×104 viable CFU of H37Rv M. tuberculosis by intratracheal route. Thirty days after challenge, the animals were sacrificed and the bacterial burden was determined by counting the number of CFU in the lungs. Lung histological sections were also analyzed. Splenic cells from primed animals produced more IL-5 but less IFN-gamma than non-primed ones. Also, prior contact with M. avium determined higher production of IgG1 and IgG2a anti-hsp65 antibodies in comparison to control groups. However, this higher immune response did not decrease the bacterial burden in the lungs. In addition, prior sensitization with M. avium decreased the parenchyma preservation observed in the group immunized only with pVaxhsp65. These results indicate that environmental mycobacteria can interfere with immunity and protective efficacy induced by DNAhsp65.

Efeitos da luminosidade no crescimento de mudas de Caesalpinia ferrea Mart. ex Tul. (Leguminosae, Caesalpinoideae)

Caesalpinia ferrea is a species used a lot as a medicinal plant, for urban arborization and landscape design in the state of Amapa. Yet there is not much ecophysiological information available on it. Light is an important environment factor that controls processes associates with the accumulation of dry matter, contributing thus for plant growth. The object of this research was to study the effect of different luminosity levels on the growth of seedlings of this species. The seedlings were transplanted to plastics bags containing a mixture of soil and sand (2:1), which were maintained in the full sun, under artificial shade with reduction of 50% and 70% of the luminosity and under natural shade of a closed canopy of forest. The experimental design was completely randomized with five replicates. Seedlings submitted to natural shade showed strong growth inhibition. In the full sun, the seedlings presented higher net assimilatory rate (NAR), lower shoot/root ratio (SRR) and lower leaf area ratio (LAR). The results showed little difference in the growth and biomass allocation between seedlings kept in 50 and 70% shade; the seedlings submitted to this treatment presented higher values of SRR and LAR. This indicates plasticity which influences a possible increase in light capture and is important, therefore, to keep the seedling growth and survival under low light levels. The results as a whole showed morphological and physiological adjustment to different light levels in Caesalpinia ferrea.

Myotoxic phospholipases A2 isolated from Bothrops brazili snake venom and synthetic peptides derived from their C-terminal region: Cytotoxic effect on microorganism and tumor cells

This paper reports the purification and biochemical/pharmacological characterization of two myotoxic phospholipases A2 (PLA2s) from Bothrops brazili venom, a native snake from Brazil. Both myotoxins (MTX-I and II) were purified by a single chromatographic step on a CM-Sepharose ion-exchange column up to a high purity level, showing Mr ∼ 14,000 for the monomer and 28,000 Da for the dimer. The N-terminal and internal peptide amino acid sequences showed similarity with other myotoxic PLA2s from snake venoms, MTX-I belonging to Asp49 PLA2 class, enzymatically active, and MTX-II to Lys49 PLA2s, catalytically inactive. Treatment of MTX-I with BPB and EDTA reduced drastically its PLA2 and anticoagulant activities, corroborating the importance of residue His48 and Ca2+ ions for the enzymatic catalysis. Both PLA2s induced myotoxic activity and dose-time dependent edema similar to other isolated snake venom toxins from Bothrops and Crotalus genus. The results also demonstrated that MTXs and cationic synthetic peptides derived from their 115-129 C-terminal region displayed cytotoxic activity on human T-cell leukemia (JURKAT) lines and microbicidal effects against Escherichia coli, Candida albicans and Leishmania sp. Thus, these PLA2 proteins and C-terminal synthetic peptides present multifunctional properties that might be of interest in the development of therapeutic strategies against parasites, bacteria and cancer. © 2008 Elsevier Inc. All rights reserved.

Effect of plastic film mulch on the production of butterhead lettuce cultivars under protected cultivation

The study was performed in a greenhouse with the aim of evaluating the production of butterhead lettuce cultivars with and without plastic film mulch in the summer. The cultivars studied were 'Elisa', 'Lídia', 'Herman' and 'Luísa'. The experimental design was a split plot, in which the main plot was plastic film mulch (with or without black polyethylene film) and the subplots were the cultivars, with three repetitions. The seedlings were grown in Styrofoam trays of 288 cells and transplanted 25 days after seeding in a space of 25 × 25 cm. Harvests were carried out 45 days after transplanting and the following variables were determined: fresh and dry weights of the aerial part, number of leaves per plant, plant height and the longitudinal and transverse diameters of the plant. Significant differences were found only among the cultivars for the characteristics studied, with no cover × cultivar interaction. Fresh weight of the aerial part of 'Lídia' (270.9 g) was greater than that of 'Herman' (142.5 g), but did not differ from those of 'Luísa' (229.8 g) or 'Elisa' (204.8). The dry weight of the aerial part of 'Lídia' (12.4 g) differed statistically only with respect to that of 'Herman' (7.9 g). The cultivars 'Luísa' and 'Elisa' had an aerial part dry weight of 11.4 and 11.2 g, respectively. The number of leaves per plant of the cultivars 'Luísa' (45.1) and 'Elisa' (43.0) was higher than for 'Herman' (24.05), but did not differ from that of the cultivar 'Lídia' (39.5). The mean transverse and longitudinal diameters, as well as the mean height of the plants, showed that 'Lídia', 'Elisa' and 'Luísa' had the greatest values. The cultivars 'Lídia', 'Elisa' and 'Luísa' were the most productive and plastic film mulch had no effect on lettuce production under protected cultivation.

Mutagenic effect of native species of the Brazilian Cerrado with anti-ulcerogenic activity

Neea theifera Oerted (Nyctaginaceae), Guapira noxia Linn. (Nyctaginaceae) and Hancornia speciosa Gomes (Apocynaceae) are plant species found in Brazilian Cerrado used popularly for the treatment of gastric ulcers. Here they are assessed for mutagenic activity by analysis of the reverse mutations induced in Salmonella typhimurium strains TA100, TA98, TA102 and TA97a, by extracts of the plants, with and without metabolic activation. Methanol and chloroform extracts of N. theifera and G. noxia and methanolic and aqueous extracts of H. speciosa were tested at five different concentrations. It was found that only the methanolic extract of H. speciosa exhibited a positive mutagenic effect, on strains TA98 and TA100 in the absence of metabolic activation. The phytochemical analysis of the species suggested that condensed tannins are the main compounds responsible for the observed effect.

Effect of the density and of the distance of Brachiaria decumbens staff on the initial growth of Coffea arabica L. Seedligns

The aim of this work was to evaluate the interference of three densities (4, 8 and 16 plants/m 2) of Brachiaria decumbens Stapf distributed at three different distances (0, 10 and 20 cm from coffee plants) on the initial growth of Coffea arabica L. seedlings. Dark red Latosoil was used as substrate to fill up 70 liters cement boxes. Coffee seedlings at 15 cm height and 8 leaves stage were planted in each box. B. decumbens seedlings were transplanted when they reached two to four leaves stage. The experimental design was a randomized blocks with ten treatments and three replications. At the end of the experiment were measured, height, leaf number, leaf area, chlorophyll content and dry weight of coffee plants. Dry weight of B. decumbens leaves was also evaluated. The leaf area and leaf dry weight was shown to be more sensitive parameters to the interference of Brachiaria plants. Dry weight of coffee plants were reduced from 60% to 88% when the B. decumbens were transplanted 0 cm from the coffee plant. With distance of 10 cm the reduction of dry weight increased with the increase of the density of the B. decumbens. In the coexistence of the coffee plants with 16 plants/m 2 of B. decumbens, there was a reduction in the dry weight of about 80% at the distances of 0 and 20 cm. Regarding leaf area, the reduction was 47% on the average in the density of 4 plants/m 2 of B. decumbens The reduction in leaf area at other densities was higher at the distance of 0 and 20 cm, and 55% reduction was obtained at the distance of 10 cm.

Soluções nutritivas no desenvolvimento do crisântemo cultivado em vaso

Although the information about nutritive solution for potted chrysanthemum is still incongruent, the objective of this work was to evaluate the effect of nutritive solutions in the development of Miramar chrysanthemum cultivated in pots at FCAV-UNESP, Jaboticabal-SP. The experiment had a randomized block design with split-plot evaluation and treatments corresponding to four nutritive solutions consisting of commercial fertilizers (S1, S2, S3, S4) evaluated at 0, 14, 28, 42, 56 and 70 days after rooting (DAE), with five replications. The plant height, leaf area, number of leaves, stem diameter, total dry matter mass of aerial part and roots; number and diameter of inflorescences, and physiological indexes of growth analysis were evaluated. The nutritive solutions did not interfere significantly in stem diameter, number and diameter of inflorescences and dry matter of roots, although S3 and S4 provided the highest plants, leaf area, number of leaves, total dry matter of aerial part. The physiological indexes showed that plants presented an accelerated growth from 14 to 28 DAE with S4 and S3 solutions and therefore they were considered the best solutions.

Effect of nozzle angle and air-jet parameters in air-assisted sprayer on biological effect of soybean asian rust chemical protection

Aiming at improving the efficiency control of Phakopsora pachyrhizi, this research evaluated different application techniques, using spray deposits and yield parameters of soybean crop. Two experiments were carried out in the experimental area of FCA/UNESP - Botucatu, SP, Brazil, in the soybean crop, Conquista variety, in the 2006/2007 season. The first experiment was arranged in random blocks with eight treatments and four replications. The treatments were conducted in factorial arrangement 4×2 (four air levels 0, 9, 11 and 29 km/h combined at two nozzle angles 0 and 30°) using AXI 110015 nozzles. Ten plants on each plot were selected for sampling spray deposits. Artificial targets were fixed on plants, two in the top and another two in the bottom part of plants (abaxial and adaxial leaf surface each one). For deposit evaluations, a cupric tracer was used and the amount of deposits was determined by a spectrophotometer. The second experiment was carried out in the same place and the treatments were of the same arrangement as the previous experiment, including control treatment (untreated plants). The spraying with triazole fungicide was realized in R2 and R5.2 growth stages of soybean with 142 l/ha spray volume. The nozzle angled of 30° combined with maximum air speed promoted the highest spray deposits on the soybean crop and influenced positively the control of the soybean Asian rust as well in the productivity of this crop.

Effect of fungicides on the mite fauna of prunus persica L. cultivars in Presidente Prudente, SP, Brazil

The objective of this study was to characterize the diversity of phytophagous and predatory mites on peach cultivars either with or without fungicide treatment, in the city of Presidente Prudente, State of São Paulo, Brazil. In order to evaluate the effect of fungicides, leaf samples were colleted at random from treated and untreated plants of the Tropical, Aurora 1 and Aurora 2 cultivars, from June 2004 to February 2006. From the results obtained it can be concluded that: plants with or without treatment showed high populations of predatory mites, indicating that the treatments were innocuous on the mite populations. A diversified composition of the mite community was observed. The Phytoseiidae family had the highest richness in numbers and species of mites. Euseius citrifolius and E. concordis were the most abundant species. Plants with or without treatment had high abundances of predatory mites, with a predominance of E. citrifolius.

Use of Physiological Parameters in Screening Drought Tolerance in Sugarcane Genotypes

The physiological response of four commercial sugarcane genotypes to water stress was evaluated by measuring the photochemical efficiency of the photosystem II (chlorophyll a fluorescence ratio, F v/F m), estimated chlorophyll content (SPAD unit), leaf temperature (LT) and leaf relative water content (RWC). A field trial was established in the subtropical area with well-watered and water-stressed genotypes, in completely randomized blocks with four replicates in a 4 × 2 × 3 factorial design (genotype × irrigation × evaluation date). Physiological measurements were done during a 90 day-period of formative stage of plants. The analysis of variance showed that the interaction of genotype × irrigation × evaluation date had a significant effect for three physiological markers tested, F v/F m, SPAD unit and RWC. Under non-stressed conditions, all genotypes showed similar responses for the four markers. Under water deficiency stress, two drought-tolerant genotypes, HOCP01-523 and TCP89-3505 displayed higher values for F v/F m, SPAD unit and RWC, and lower values for LT, and could be classified as tolerant. It is therefore possible to use these physiological water stress associated traits as scorable marker traits for selecting drought-tolerant sugarcane genotypes in future breeding programs. © 2011 Society for Sugar Research & Promotion.

Effect of different pulse treatments on vase life of inflorescences of oncidium varicosum 'Samurai'

Oncidium varicosum is a species of orchid that is prevalent in South America. This species produces yellow flowers, which are commonly named golden shower, ballet dancer and Anita's flower. The study of the post-harvest physiology of flowers involves examination of factors related to quality loss of cut flowers, which is related not only to several physiological processes but also to interactions between the floral components and qualitative factors. In this context, our research involved the study of physiological aspects of the post-harvest physiology of cut inflorescences of Oncidium varicosum ('Samurai') after treatment with pulsing solutions containing silver salts, 8-HQC and sucrose. A reduction of relative water content of Oncidium flowers treated with pulsing STS (2 mM) was observed. The soluble carbohydrates and reducing sugars content decreased, and higher contents were measured in flowers treated with STS (2 mM). All parameters related to flower coloration were reduced in all treatments. Similar results were obtained for carotenoid content, except for the STS treatment (2 mM), which differed significantly from the other treatments on the 8th and 12th days of vase life. The inflorescences' longevity varied from 12 days (distilled water) to 15 days (STS 2 mM).

Desiccation tolerance in seeds of Annona emarginata (Schldtl.) H. Rainer and action of plant growth regulators on germination

Annona emarginata (Schldtl.) H. Rainer (araticum-de-terra-fria) is used as a rootstock for several species of Annonaceae. It is suggested that these seeds should be sown immediately after extraction and, therefore, they could be intolerant to desiccation. There are several mechanisms involved with desiccation tolerance. Soluble sugars, for example, can accumulate and act as osmoprotectants for the membrane system during desiccation. The aim of this study is to assess desiccation tolerance in A. emarginata seeds. In addition, we examined the soluble sugars involved in desiccation tolerance. Finally, we determined the effect of gibberellic acid (GA4+7) and N-(phenylmethyl)-aminopurine in promoting the germination of seeds with different water contents. The experiment consisted of a randomized 4×5 factorial design (desiccation levels × concentration of growth regulators). After drying, seeds containing 31 (control), 19, 12 and 5% water were incubated in different concentrations of GA4+7 N-(phenylmethyl)-aminopurine (0, 250, 500, 750 and 1000 mg L-1) for 60 hours. The experiment was conducted in a germination chamber with alternating temperature and photoperiod of 20oC for 18 hours of darkness and 30oC for 6 hours of light. We analyzed electrical conductivity, germination rate, mean germination time, germination speed, frequency and uniformity of germination, percentage of dormant seeds and soluble sugar profile in intact seeds through high-performance liquid chromatography (HPLC). The data were subjected to analysis of variance, and the means were compared using Tukey's test at a threshold of p<0.05. The results showed that seeds of A. emarginata appears to be desiccation tolerant and, also, that sucrose increases when seed water content is reduced to values as low as 12%, exogenous GA4+7+N-(phenylmethyl)-aminopurine improves its germinability.

Genotoxicity of the medicinal plant Maytenus robusta in mammalian cells in vivo.

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

EFFECT OF DIFFERENT ROOTSTOCKS ON GROWTH, CHLOROPHYLL A FLUORESCENCE AND MINERAL COMPOSITION OF TWO GRAFTED SCIONS OF TOMATO

Two tomato scions (cvs. 'Raf' and 'Gorety') were grafted on three different rootstocks: S. torvum, 'Beaufort' (Lycopersicum esculentum × Lycopersicum hirsutum) and intermediate grafting of eggplant 'Cristal' between tomato and S. torvum (double graft). Plants were grown in Mediterranean greenhouse conditions. The response to grafting was measured through growth parameters, Fv/Fm and leaf macronutrients analysis, and it was compared with non-grafted plants. The scions grafted on S. torvum in simple and double graft showed lower fresh and dry weight of leaves, number of commercial fruits, plant height, Fv/Fm and decreased their capacity to absorb several nutrients resulting in a lower mineral concentration in scions leaves, as a result of a thickened graft union. On the other hand, both scions showed a good response when grafted on the rootstock 'Beaufort', with which growth parameters, yield and photosynthetic capacity were similar to non-grafted plants. © 2013 Copyright Taylor and Francis Group, LLC.

Gastroprotective effect of the ethanolic extract and fractions obtained from syngonanthus bisulcatus Rul

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