Toxoplasma gondii: Evaluation of an intranasal vaccine using recombinant proteins against brain cyst formation in BALB/c mice

The purpose of this work was to evaluate protective activity against brain cyst formation in BALB/c mice intranasally vaccinated with recombinant proteins from Toxoplasma gondii. The recombinant proteins rROP2, rGRA5 and rGRA7 were used in vaccine preparation. Thirty-three female mice were divided into three groups, these animals received two doses by intranasal route at days 0 and 21 as follows; group 1 (G1, n = 11) received 12.5 mu g of each recombinant protein plus 0.5 mu g of cholera toxin, group 2 (G2, n = 11) received phosphate buffer saline (PBS) plus 0.5 mu g of cholera toxin, and group 3 (G3, n = 11) received PBS only. At challenge day (day 33) three animals from each group were euthanatized for IgA measure from intestine. Mice were infected orally with 50 cysts from the VEG strain at day 33. At challenge day the G1 animals had high immunoglobulin A levels, however, they only showed IgG antibody titers against rROP2 and rGRAT Animals from G1 also exhibited strong resistance to cyst formation compared with the control group (G3, P < 0.05). However, we did not observe differences in protection against brain cyst formation between G1 and G2 (P > 0.1). These results indicate that intranasal immunization in BALB/c mice with recombinant proteins rROP2, rGRA5 and rGRA7 associated with cholera toxin induced partial protection, when compared with G3, against tissue cyst formation after oral infection with tissue cysts from T gondii. (c) 2007 Elsevier B.V. All rights reserved.

Effect of wounds applied to the bases of herbaceous cuttings on the rooting of four Japanese apricot clones (Prunus mume Sieb. et Zucc.) in an intermittent mist system

The Japanese apricot (Prunus mume Sieb. et Zucc.) is a fruit tree of the Rosaceae family which produces very acid and bitter fruits, highly appreciated by Orientals. In Brazil, this species has been studied as a rootstock for peach and nectarine trees, its main advantage being the reduction in plant vigour, which can favour the production of compact trees and orchard cultural treatments. This study was conducted in the Vegetable Production Department of FCAV/UNESP, Jaboticabal Campus, São Paulo State, Brazil, and the objective was to examine the effect of wounding the herbaceous cutting bases on the rooting of four Japanese apricot clones. The clones were obtained from plants under cultivation in the Instituto Agronomico de Campinas, Brazil, and were identified as Clones 02, 05, 10 and 15. The stock plants, obtained through herbaceous cuttings, were maintained under lath house conditions (50% of natural light). Cuttings 12 cm long with 3 to 5 leaves were collected from these clones and prepared. The experiment was carried out in a completely randomised design with 4 repetitions of 20 cuttings per replication, in a factorial 4 x 2 design, the clone factor having 4 levels (Clones 02, 05, 10 and 15) and the wounding factor at 2 levels of incisions into the cutting base (with and without). All the cuttings were treated with 2000 mg.L-1 of IBA for five seconds. Differences between the clones were observed concerning the rooting percentage, dead cuttings, number and length of roots. The incision (wound) at the base of the herbaceous cuttings of the Japanese apricot increased the number of roots and improved the distribution of these in the damaged tissue but the results were not considered sufficiently beneficial to make the treatment worthwhile.

Efeito do comprimento de estacas herbáceas de dois clones de umezeiro (Prunus mume Sieb & Zucc.) no enraizamento adventício

O umezeiro (Prunus mumeSieb & Zucc.) é uma rosácea de folhas caducas, nativa da China, cujos frutos e flores são muito apreciados pelos povos orientais. No Brasil, alguns estudos foram realizados visando a sua utilização como porta-enxerto para pessegueiro e nectarineira, dadas as suas características de adaptação, rusticidade, redução do porte da planta e compatibilidade com algumas cultivares de Prunus persica. O presente estudo foi conduzido em câmara de nebulização sob ripado, pertencente ao Departamento de Produção Vegetal da FCAV/UNESP, Câmpus de Jaboticabal-SP. Objetivou-se verificar a influência de quatro comprimentos de estacas herbáceas no enraizamento de dois clones de umezeiro. O material vegetal, identificado como Clone 10 e Clone 15, foi oriundo do Programa de Melhoramento Genético do Instituto Agronômico de Campinas-SP. O experimento foi constituido de fatorial 2 x 4, em blocos casualizados, sendo o fator clone em 2 níveis (Clone 10 e Clone 15) e o fator comprimento de estaca em 4 níveis (12; 15; 18 e 25cm). Pelos resultados observados, verificou-se diferença entre os clones somente na porcentagem de estacas brotadas e número de raízes por estaca. O comprimento da estaca influenciou na porcentagem de enraizamento e na mortalidade das estacas, sendo que estacas maiores tenderam a apresentar maiores porcentagens de enraizamento e menores de mortalidade. As estacas com 12cm, embora apresentando menor número de raízes por estaca, são recomendadas por permitirem a obtenção de um maior número de estacas por planta-matriz. Houve efeito significativo da interação entre os fatores para número e comprimento de raízes.

Desenvolvimento inicial no campo de pessegueiros 'Aurora-1' enxertados em clones de umezeiro e 'Okinawa' propagados por estacas herbáceas

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

Enraizamento de estacas herbáceas de quatro clones de umezeiro (Prunus mume Sieb. et Zucc.) durante o inverno ameno, em Jaboticabal-SP

Estudos realizados no Brasil com o umezeiro (Prunus mume Sieb. et Zucc.) relatam promissoras perspectivas de utilização desta espécie como porta-enxerto para pessegueiro e nectarineira, em função de sua rusticidade, adaptação ao inverno brando, compatibilidade com Prunus persica, redução do vigor das plantas e melhoria da qualidade dos frutos. Entretanto, em função da propagação por sementes, tem sido observadas diferenças de vigor entre as plantas, resultando em pomares muito heterogêneos. Assim, o presente estudo teve por objetivo estudar o enraizamento de estacas herbáceas de quatro clones de umezeiro (Clones 02, 05, 10 e 15) durante o inverno ameno, em Jaboticabal-SP. O experimento foi conduzido entre os meses de junho e agosto, sendo avaliado aos 70 dias após a estaquia. Pelos resultados obtidos, foi possível concluir que é viável a propagação dos clones estudados por enraizamento de estacas herbáceas durante o inverno. Foram observadas diferenças entre os clones quanto à porcentagem de enraizamento, porcentagem de estacas com calo, número e comprimento das raízes. No conjunto das variáveis analisadas, os melhores resultados foram obtidos com os Clones 10 e 15.

Conditions affecting production of functional muscle recombinant alpha-tropomyosin in Saccharomyces cerevisiae

Yeasts are attractive hosts for heterologous protein production as they follow the general eukaryotic post-translational modification pattern. The well-known Saccharomyces cerevisiae has been used to produce a large variety of foreign proteins. The proper function of muscle tropomyosin depends on a specific modification at its N-terminus. Although tropomyosin has been produced in different expression systems, only the recombinant protein produced in the yeast Pichia pastoris has native-like functional properties. In this paper we describe the production of functional skeletal muscle tropomyosin in the yeast S. cerevisiae. The recombinant protein was produced in high amounts and production was strongly affected by genetic and environmental factors, including plasmid copy number, promoter strength, and growth media composition. (C) 2003 Elsevier B.V. (USA). All rights reserved.

A novel system for large-scale gene expression analysis: bacterial colonies array

In the present work, we report the use of bacterial colonies to optimize macroarray technique. The devised system is significantly cheaper than other methods available to detect large-scale differential gene expression. Recombinant Escherichia coli clones containing plasmid-encoded copies of 4,608 individual expressed sequence tag (ESTs) were robotically spotted onto nylon membranes that were incubated for 6 and 12 h to allow the bacteria to grow and, consequently, amplify the cloned ESTs. The membranes were then hybridized with a beta-lactamase gene specific probe from the recombinant plasmid and, subsequently, phosphorimaged to quantify the microbial cells. Variance analysis demonstrated that the spot hybridization signal intensity was similar for 3,954 ESTs (85.8%) after 6 h of bacterial growth. Membranes spotted with bacteria colonies grown for 12 h had 4,017 ESTs (87.2%) with comparable signal intensity but the signal to noise ratio was fivefold higher. Taken together, the results of this study indicate that it is possible to investigate large-scale gene expression using macroarrays based on bacterial colonies grown for 6 h onto membranes.

Effect of recombinant bovine somatotropin (bST) on follicular population and on in vitro buffalo embryo production

The objective of this study was to evaluate the effect of bovine somatotropin (bST) on ovarian follicular population in buffalo heifers and its influence on oocyte quality, recovery rates and in vitro embryo production. We tested the hypothesis that bST treatment in buffalo females submitted to an ovum pick-up (OPU) program Would improve the number of follicles recruited, oocyte quality and in vitro embryo production. A total of 10 heifers were assigned into two treatment groups: group bST (n = 5; receiving 500 mg of bST in regular intervals) and control group (n = 5; without additional treatment). Both groups were subjected to OPU sessions twice a week (every 3 or 4 days), for a total of 10 sessions per female, although due to procedural problems, only the first five OPU sessions produced embryos. The number of follicles and the diameters were recorded at all OPU sessions. The harvested oocytes were counted and classified according to their quality as either A, B, C, D or E, with A and B considered good quality. Cleavage and blastocyst production rates were evaluated 2 and 7 days after in vitro fertilization, respectively. The bST treatment increased the total number of antral follicles (> 3 mm in diameter; 12.2 compared with 8.7: p, < 0.05) and of small antral follicles (< 5 mm; 9.1 compared with 6.5; p < 0.05) per OPU session. The bST also tended to increase the number of oocytes recovered per session (5.2 compared with 4.1; p = 0.07), and enhanced the percentage of good quality oocytes (48.8% compared with 40.6%; p = 0.07), bST showed no effect on cleavage and blastocyst production rates (p > 0.05). The significant effects of performing repeated OPU sessions were decreasing the follicular population (p < 0.001) as well as the number of follicles aspirated (p < 0.001), and oocytes recovered (p < 0.02). In conclusion, bST treatment improves the follicular population, demonstrating its possible application in buffalo donors submitted to OPU programs. (c) 2008 Elsevier B.V. All rights reserved.

Influência da densidade básica da madeira de clones de Eucalyptus grandis x Eucalyptus urophylla na qualidade da polpa branqueada.

The study analyzed the wood basic density effect in two Eucalyptus grandis x Eucalyptus urophylla hybrid clones (440 kg/m(3) e 508 kg/m(3)) on bleached pulp quality (fiber dimensions and physical-mechanical properties). The woods performance on pulping, bleaching and beating results were analyzed. The Kraft pulping was carried out in forced circulation digester in order to obtain 17 +/- 1 kappa number targets. The pulps were bleached to 90 +/- 1 using delignification oxygen and D0EOPD1 bleaching sequence. Bleached pulp of low basic density clone showed, significantly, lowest revolutions number in the PFI mill to reach tensile index of 70 N.m/g, low Schopper Riegler degree and generated sheets with higher values to bulk and opacity. These characteristics and properties allow concluding that bleached pulp of low basic density clone was the most indicated to produce printing and writing sheets. The bleached pulp of high basic density clone showed higher values of bulk and capillarity Klemm and lower water retention value when analyzed without beating. The bleached pulp of high basic density clone showed more favorable characteristics to the production of tissue papers.

A naturally occurring recombinant isolate of Lettuce mosaic virus

LMV-Common and LMV-Most are two seed-borne types of Lettuce mosaic virus (LMV), genus Potyvirus. LMV-Most, but not LMV-Common, overcomes the resistance afforded to lettuce by two recessive genes, mo1(1) and mo1(2). An RT-PCR-based assay thought to be specific for LMV-Most also amplified LMV-Tn2, previously typified as LMV-Common. The sequence of selected regions along the genome indicated that LMV-Tn2 is a natural recombinant between LMV-Most and LMV-Common isolates, with a putative recombination site located within the P3 coding region. This is the first evidence of a naturally occurring LMV recombinant isolate.

Densidade básica da madeira de sete espécies e três clones de eucalipto antes e durante o cultivo de shiitake

Avaliou-se a densidade básica da madeira e casca de sete espécies (E. saligna, E. grandis, E. urophylla, E. camaldulensis, E. citriodora, E. paniculata e E. pellita) e três clones de eucalipto (híbridos de E. grandis x E. urophylla) antes e durante o cultivo das linhagens LE-95/01 e LE-96/18 de shiitake (Lentinula edodes) em toras. Cada linhagem de shiitake foi inoculada em nove toras de cada tipo de eucalipto com 1 m de comprimento e 9 a 14 cm de diâmetro. Assim, o delineamento experimental foi inteiramente casualizado, com 20 tratamentos e 9 repetições, sendo cada repetição correspondente a uma tora. As toras foram mantidas em estufa climatizada, com temperatura de 25 ± 5ºC e umidade relativa do ar entre 60-80% durante 12 meses. Para a determinação da densidade básica, analisaram-se cunhas de discos e cascas de eucalipto recém-cortadas (sem inoculação das linhagens de L. edodes) e em cunhas de discos retirados de toras já inoculadas com as linhagens de L. edodes após 8 e 12 meses de incubação. Verificou-se que a densidade básica da madeira, ao longo do ciclo de cultivo, foi reduzida em todos os tipos de eucalipto.

Seletividade e absorção radicular do sulfentrazone em clones de eucalipto

O objetivo deste trabalho foi avaliar a seletividade e a absorção do sulfentrazone em clones de eucalipto. O primeiro experimento foi instalado em casa de vegetação, em delineamento inteiramente casualizado, com quatro repetições, no esquema fatorial 2 x 4, sendo duas doses do sulfentrazone (400 e 600 g ha-1) e quatro clones de eucalipto, híbridos de Eucalyptus grandis x E. urophylla (FB1, FB2, FB3 e FB4). Foram realizadas avaliações visuais de intoxicação das plantas de eucalipto e, no final do estudo, determinou-se a massa seca da parte aérea dos clones. No segundo experimento, foram utilizados os mesmos clones, sendo estes acondicionados em tubos falcon com 50 mL da solução contendo o sulfentrazone na concentração de 129 mM. As plantas de eucalipto permaneceram por 24 horas com as raízes imersas na solução e, em seguida, foi realizada a extração da seiva do xilema das plantas por meio de uma câmara de pressão. A concentração de sulfentrazone na seiva das plantas foi determinada através de cromatografia líquida e espectrometria de massas. O clone FB3 apresentou menor acúmulo de massa seca em relação aos demais, o que pode estar diretamente associado aos altos níveis de intoxicação observados. O clone FB2, apesar de mostrar elevada intoxicação, não apresentou níveis tão elevados de redução de massa seca em relação à testemunha. No tocante às concentrações de sulfentrazone nas plantas, elas foram proporcionais ao acúmulo de massa seca, o que indica que as variações na seletividade dos clones de eucalipto podem estar relacionadas à absorção diferenciada do herbicida.

Intensidade do mal das folhas em plantas jovens e adultas de seis clones de seringueira na região do Vale do Ribeira: South American leaf blight intensity evaluated in six clones of young and adult rubber trees in the Vale do Ribeira region, São Paulo state, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo comparativo da constituição nutricional da madeira e casca de espécies e clones de Eucalipto visando o cultivo de Shiitake em toras

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Clones promissores de seringueira para a região do Vale do Ribeira, São Paulo

Este trabalho objetivou apresentar resultados da primeira seleção de clones de seringueira (Hevea brasiliensis (Willd. ex Adr. de Juss.) Muell.Arg.) da série IAC 300, e amazônicos das séries IAN e Fx, em experimento de pequena escala, visando produção, crescimento e resistência ao mal-das-folhas. A produção e o vigor de 20 clones foram avaliados por dois e nove anos, respectivamente, em PariqüeraAçú, no Vale do Ribeira, SP. Os clones IAC 301, IAC 304, IAC 306 e IAC 319 produziram satisfatoriamente nos dois primeiros anos de sangria. Os clones amazônicos IAN 6323, Fx 3864 e IAN 2903, com produções de 1.078 kg, 945 kg e 900 kg/ha/ano, respectivamente, foram superiores à testemunha IAN 873 (878 kg/ha/ano). Os clones selecionados apresentaram crescimento vigoroso, com extremos de perímetro do caule, na abertura do painel, de 37,40 cm (IAN 4493) a 53,75 cm (IAN 6323), e percentual de plantas aptas à sangria de 7,0% (IAN 4493) a 100% (IAN 6323 e IAC 302), exceto os clones Fx 3899 e IAN 3044. O IAC 315, com 7,37 mm, mostrou maior espessura de casca virgem que o IAN 873 (6,44 mm). Os clones IAC 320, IAC 306 e IAC 315 foram os mais resistentes ao Microcyclus ulei.