ABSTRACT The objective of this work was to characterize reference bacteria strains, belonging to the genus Bacillus and species of correlated genera, by simplified morphological and biochemical methods. The morphological characterization is based on the aspects of the colonies, as well as cytomorphology of the species, by optical and scanning microscopy. For biochemical characterization, the sensitivity test to antimicrobials by disk-diffusion is performed. Moreover, the strains were characterized by extracting intracellular proteins. Characteristics such as shape, color, and consistency of the colonies, in addition to the type of spore and production of protein crystals were determinants for the morphological characterization of these species. The antibiogram revealed high resistance to β-lactam group antibiotics, in species of Bacillus cereus s.l group. In Bacillus subtilis s.l. group there was high susceptibility to antibiograms, mainly for species of B. subtilis. The protein profile provided specific protein patterns for some species, mainly bands of 130 e 65 kDa for B. thuringiensis, 140 e 130 kDa for Lysinibacillus sphaericus, and 115 kDa for Brevibacillus laterosporus. Our results showed that the morphological and biochemical characterizations, provided a simple identification, with easy interpretation, and low cost. genera methods colonies microscopy diskdiffusion disk diffusion performed Moreover proteins shape color βlactam β lactam antibiotics sl s l antibiograms B 13 6 thuringiensis 14 sphaericus 11 laterosporus characterizations identification interpretation cost 1

Abstract Anatomical and histochemical studies were carried out during the somatic embryogenesis of Syagrus oleracea (Mart.) Becc. from immature inflorescences. Immature rachillas were inoculated on Murashige and Skoog medium with 4.52 μM 4-amino-3,5,6-trichloropicolinic acid (Picloram) and 2,4-diclorophenoxyacetic acid. Auxin concentrations were gradually reduced for the formation of somatic embryos, while for plantlet regeneration, embryos were inoculated on a medium without growth regulator. After 270 days, two types of embryogenic calluses were observed in explants under the effect of Picloram: nodular, with a smooth surface and those without definite shape, with an irregular surface, both presenting a meristematic zone with polysaccharide content. Callus progression to the differentiation stage allowed for the conversion of somatic embryos, some with a well-defined protoderm and procambial bundles. The histochemical analysis of the somatic embryos did not identify the presence of polysaccharide and protein reserves, which emphasizes the necessity for optimization of the maturation conditions.

ABSTRACT Brachiaria brizantha is a forage grass well adapted to tropical areas and cultivated in millions of hectares in Brazil. The apomictic mode of reproduction in this species, in addition to differences in ploidy between sexual and apomictic plants, impairs crossbreeding. The development of a methodology to transform apomictic cultivars will provide an option to introduce agronomic important traits to B. brizantha cv. Marandu. In addition, it will open the possibility to study in vivo the function of candidate genes involved in the apomictic reproduction. The objective of this work was to evaluate peeled seeds, isolated embryo from mature seeds, embryogenic calluses and embryogenic cell suspensions, as target explant for genetic transformation via biolistics. Plasmids bearing the marker genes gus and hptII under the control of the rice actin 1 promoter (pAct1-Os) or the maize ubiquitin 1 promoter (pUbi1Zm) were used. All the target-explants used were suitable for transient gene expression after bombardment, showing gus expression and resistance to hygromycin. Using embryogenic calluses and cell suspensions as target tissues, transgenic plants were regenerated and transgenes detected.

O clima organizacional, um dos construtos de maior complexidade na área organizacional, apresenta características que podem ser percebidas de formas distintas por diferentes profissionais. Este estudo teve como objetivo desenvolver e validar a Escala de Clima Organizacional para Organizações de Saúde. Para validar o instrumento foram investigados 668 trabalhadores da área de saúde, que responderam a 100 itens, distribuídos em 13 dimensões. Foi utilizada para estudo da validade a análise fatorial e, para exame da consistência interna, o cálculo do alpha de Cronbach. Os resultados apontaram como melhor solução fatorial a estrutura de 64 itens, dispostos em sete fatores. A consistência interna da medida foi de 0,934. A Escala de Clima Organizacional para Organizações de Saúde demonstrou boa qualidade psicométrica, considerando-se o contexto cultural investigado. Sugere-se, entretanto, a ampliação dos estudos para outros tipos de amostra e organizações, de modo a verificar a validade transcultural do construto.

Organizational climate, as one of the most complex constructs in the organizational area, possesses characteristics that may be perceived in distinct ways by different professionals. This study aimed to develop and validate the Organizational Climate Scale for Health Organizations. In order to validate the tool, 668 health workers were investigated, responding to 100 items, distributed across thirteen dimensions. Factorial analysis was used to validate and calculate Cronbach's alpha in order to estimate internal consistency. The results suggest the best factorial solution was a structure composed of 64 items, organized into seven factors. The internal consistency was 0.934. The Organizational Climate Scale for Health Organizations shows good psychometric quality, considering the cultural context investigated. However, it is recommended that these studies be extended to other types of samples and organizations, in order to verify the cross-cultural construct validity.

O objetivo deste trabalho foi investigar prováveis modos de ação da levedura Cryptococcus magnus, que resultam no controle da antracnose (Colletotrichum gloeosporioides) em frutos de mamoeiro na póscolheita. A microscopia eletrônica de varredura foi utilizada para avaliar o efeito da levedura sobre inoculações realizadas após a colheita. Os resultados mostraram que C. magnus é capaz de colonizar a superfície de ferimentos nos frutos e vencer a competição por espaço e, provavelmente, por nutrientes. Além disso, C. magnus produz uma matriz de textura característica que afeta a integridade da hifa do patógeno. A competição por espaço e a produção de substâncias que afetam a integridade das hifas estão entre os mais importantes modos de ação desta levedura.

The objective of this work was to investigate possible modes of action of the yeast Cryptococcus magnus in controlling anthracnose (Colletotrichum gloeosporioides) on post harvested papaya fruits. Scanning electron microscopy was used to analyze the effect of the yeast on inoculations done after harvest. Results showed that C. magnus is able to colonize wound surfaces much faster than the pathogen, outcompeting the later for space and probably for nutrients. In addition, C. magnus produces a flocculent matrix, which affects hyphae integrity. The competition for space and the production of substances that affect hyphae integrity are among the most important modes of action of this yeast.

O objetivo deste trabalho foi selecionar e caracterizar, no Banco de Germoplasma de Bacillus spp., da Empresa Brasileira de Pesquisa Agropecuária, as estirpes de Bacillus thuringiensis mais tóxicas à Plutella xylostella, por métodos morfológicos, bioquímicos e moleculares. Das 203 estirpes testadas, sete causaram 100% de mortalidade e foram semelhantes à estirpe padrão utilizada, B. thuringiensis subsp. kurstaki. Elas apresentaram proteínas de 130 kDa e 65 kDa, presença de genes cry1 e cry2 e cristais bipiramidais, cubóides e redondos. As estirpes selecionadas oferecem novas perspectivas de controle de P. xylostella.

The aim of this work was to select and characterize the most toxic Bacillus thuringiensis strains, from the Germplasm Bank of Bacillus spp. of Empresa Brasileira de Pesquisa Agropecuária, against Plutella xylostella. Strains were characterized by morphological, biochemical and molecular methods. It was observed that seven out of the 203 strains tested showed high toxicity compared to the standard used B. thuringiensis subsp. kurstaki (HD-1), which showed 100% mortality. Selected strains showed features described for lepidoptera regarding the protein of 130 kDa and 65 kDa; profile and features were obtained through the PCR reactions, making possible to identify the presence of cry1 and cry2 genes. Moreover, the scanning electron microscopy showed the bipiramydal, cubed and round crystal forms. The selected strains offer new perspectives to control P. xylostella.

Neste trabalho foram realizados testes de patogenicidade com 210 estirpes de Bacillus thuringiensis contra larvas de Aedes aegypti e Culex quinquefasciatus, a fim de se determinar as mais eficazes. Estas estirpes foram isoladas de diversas regiões do Brasil e estão armazenadas na coleção de Bacillus spp. da Embrapa Recursos Genéticos e Biotecnologia. As estirpes selecionadas foram caracterizadas por métodos morfológicos (microscopia), bioquímicos (SDS-PAGE 10%) e moleculares (Reação em Cadeia da Polimerase). Foram selecionadas seis estirpes entomopatogênicas de Bacillus thuringiensis. Nenhuma das estirpes de Bacillus thuringiensis apresentou produtos de PCR esperados para a detecção dos genes cry4, cry11 e cyt1A. A patogenicidade das estirpes não está associada à presença das toxinas Cry4, Cry11 ou Cyt, assim, essas estirpes poderão ser utilizadas para a formatação de um bioinseticida alternativo contra mosquitos.

The objective of this work was to evaluate 210 Bacillus thuringiensis strains against Aedes aegypti and Culex quinquefasciatus larvae to select the most effective. These strains were isolated from different regions of Brazil and are stored in a Bacillus spp. collection at Embrapa Recursos Genéticos e Biotecnologia, Brasília, Brazil. The selected strains were characterized by morphological (microscopy), biochemical (SDS-PAGE 10%) and molecular (PCR) methods. Six B. thuringiensis strains were identified as mosquito-toxic after the selective bioassays. None of the strains produced the expected PCR products for detection of cry4, cry11 and cyt1A genes. These results indicate that the activity of mosquitocidal Brazilian strains are not related with Cry4, Cry11 or Cyt proteins, so they could be used as an alternative bioinsecticide against mosquitoes.

O objetivo deste trabalho foi selecionar entre 300 estirpes de Bacillus thuringiensis as efetivas simultaneamente contra larvas de Spodoptera frugiperda J.E. Smith e Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae), Anthonomus grandis Boheman (Coleoptera: Curculionidae), Aedes aegypti Linnaeus e Culex quinquefasciatus Say (Diptera: Culicidae). Foram selecionadas duas estirpes de B. thuringiensis, denominadas S234 e S997, que apresentaram atividade contra as três ordens de insetos. As estirpes foram caracterizadas por métodos morfológicos, bioquímicos e moleculares. As mesmas apresentaram duas proteínas principais de 130 e 65 kDa, produtos de reação em cadeia da polimerase de tamanho esperado para a detecção dos genes cry1Aa, cry1Ab, cry1Ac, cry1B e cry2 e cristais bipiramidais, cubóides e esféricos.

The aim of this work was to select among 300 strains of Bacillus thuringiensis those which are simultaneously effective against larvae of Spodoptera frugiperda J.E. Smith and Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae), Anthonomus grandis Boheman (Coleoptera: Curculionidae), Aedes aegypti Linnaeus and Culex quinquefasciatus Say (Diptera: Culicidae). Two strains of B. thuringiensis were selected, S234 and S997, which presented activity against those three insect orders. Both strains were characterized by morphological, biochemical and molecular methods. They have presented two main proteins with 130 and 65 kDa, polimerase chain reaction products with expected sizes for detection of the genes cry1Aa, cry1Ab, cry1Ac, cry1B and cry2 and bipiramidal, cubical and spherical crystals.

O objetivo desse trabalho foi avaliar a influência da colesterol oxidase (Coase) na eclosão e viabilidade de larvas do bicudo-do-algodoeiro (Anthonomus grandis Boheman, 1843). Uma série de bioensaios foram conduzidos com ovos e larvas neonatas, os quais foram expostos a diferentes concentrações da enzima em dieta artificial. A sobrevivência das larvas foi afetada em todas as concentrações testadas, e a DL50, no sexto dia, foi de 53 mig/mL (IC 95%: 43-59). A Coase também interferiu na eclosão das larvas quando os ovos foram submersos por 15 min em solução de Coase, em diferentes concentrações. Observações feitas ao microscópio de luz e eletrônico no intestino médio das larvas alimentadas durante seis dias com dieta artificial contendo 53 mig/mL de Coase revelou regiões altamente vacuolizadas nas células epiteliais, bem como degradação parcial das membranas basal e microvilli.

The aim of this work was to evaluate the influence of the enzyme cholesterol oxidase (Coase) on emergence and viability of larvae of the cotton boll weevil (Anthonomus grandis Boheman, 1843). A series of bioassays was performed with eggs and neonate larvae exposed to different enzyme concentrations in artificial diet. Larval survival was affected at all enzyme concentrations tested, and the six-day LD50 was 53 mug/mL (CI 95%: 43-59). Coase also interfered with hatching of larvae after eggs were floated for 15 min in Coase solution at different concentrations. Observations at the light and electronic microscopic level of midguts from larvae fed on artificial diet containing 53 mug/mL of Coase and collected at six days revealed highly vacuolated regions in the epithelial cells as well as partial degradation of the basal membrane and microvilli.

OBJECTIVE:To assess the changes in the medicamentous treatment of elderly patients hospitalized with acute myocardial infarction occurring over an 8-year period. METHODS:We retrospectively analyzed 379 patients above the age of 65 years with acute myocardial infarction who were admitted to the coronary unit of a university-affiliated hospital from 1990 to 1997. The patients were divided into 2 groups, according to the period of time of hospital admission as follows: group 1 - from 1990 to 1993; and group 2 - from 1994 to 1997. RESULTS:The use of beta-blockers (40.8%chi 75.2%, p<0.0001) and angiotensin-converting enzyme inhibitors (42% chi59.5%, p=0.001) was significantly greater in group 2, while the use of calcium antagonists (42% chi 18.5%, p<0.0001) and general antiarrhythmic drugs (19.1% chi 10.8%, p=0.03) was significantly lower. No significant difference was observed in regard to the use of acetylsalicylic acid, thrombolytic agents, nitrate, and digitalis in the period studied. The length of hospitalization was shorter in group 2 (13.4±8.9 days chi 10.5±7.5 days, p<0.001). The in-hospital mortality was 35.7% in group 1 and 26.6% in group 2 (p=0.07). CONCLUSION: Significant changes were observed in the treatment of elderly patients with acute myocardial infarction, with a greater use of beta-blockers and angiotensin-converting enzyme inhibitors and a lower use of calcium antagonists and antiarrhythmic drugs in group 2. The length of hospitalization and the mortality rate were also lower in group 2, even though the reduction in mortality was not statistically significant.