Type 21 Design B Port Side

print number 66; Initials Lower Right: ELW (Everett Longley Warner)

Relatório de estágio com vista à obtenção do grau de mestre em Educação Pré-escolar e Ensino do 1º Ciclo do Ensino Básico

Este relatório foi elaborado com vista à obtenção de grau mestre em Educação PréEscolar e Ensino do 1º Ciclo do Ensino Básico. Descreve e reflete o processo da minha intervenção junto de um grupo de crianças em idade de creche (12 e 24 meses) que frequentava o Infantário Rainha Santa Isabel. O presente trabalho tem como principal objetivo a congregação de pressupostos teóricos e metodológicos que, nesta etapa de formação, foram o ponto de partida para colocar em prática diversos pressupostos, crenças e valores. Estes, ao longo da minha formação académica foram tendo cada vez mais importância, do ponto de vista do meu desenvolvimento futuro profissional. Um outro objetivo foi pesquisar e repensar os afetos partilhados entre adulto/criança, no contexto educativo da Creche, uma vez que a primeira infância é um processo fulcral e irreversível no desenvolvimento cognitivo, social e emocional da criança. Após todo o processo reflexivo, realizando ao longo da intervenção pedagógica, foi possível verificar que o desenvolvimento e aprendizagem das crianças estão sustentados numa relação de afetos, confiança e empatia. A intervenção dos adultos e a participação das crianças dependem grandemente do ambiente emocional que é criado e recriado.

Functional characterization by genetic complementation of aroB-Encoded dehydroquinate synthase from Mycobactetium tuberculosis H37Rv and its heterologous expression and purification

The recent recrudescence of Mycobacterium tuberculosis infection and the emergence of multidrug-resistant strains have created an urgent need for new therapeutics against tuberculosis. The enzymes of the shikimate pathway are attractive drug targets because this route is absent in mammals and, in M. tuberculosis, it is essential for pathogen viability. This pathway leads to the biosynthesis of aromatic compounds, including aromatic amino acids, and it is found in plants, fungi, bacteria, and apicomplexan parasites. The aroB-encoded enzyme dehydroquinate synthase is the second enzyme of this pathway, and it catalyzes the cyclization of 3-deoxy-D-arabino-heptulosonate-7-phosphate in 3-dehydroquinate. Here we describe the PCR amplification and cloning of the aroB gene and the overexpression and purification of its product, dehydroquinate synthase, to homogeneity. In order to probe where the recombinant dehydroquinate synthase was active, genetic complementation studies were performed. The Escherichia coli AB2847 mutant was used to demonstrate that the plasmid construction was able to repair the mutants, allowing them to grow in minimal medium devoid of aromatic compound supplementation. In addition, homogeneous recombinant M. tuberculosis dehydroquinate synthase was active in the absence of other enzymes, showing that it is homomeric. These results will support the structural studies with M. tuberculosis dehydroquinate synthase that are essential for the rational design of antimycobacterial agents.

Genetic differentiation between sympatric populations of Bacillus cereus and Bacillus thuringiensis

Little is known about genetic exchanges in natural populations of bacteria of the spore-forming Bacillus cereus group, because no population genetics studies have been performed with local sympatric populations. We isolated strains of Bacillus thuringiensis and B. cereus from small samples of soil collected at the same time from two separate geographical sites, one within the forest and the other at the edge of the forest. A total of 100 B. cercus and 98 B. thuringiensis strains were isolated and characterized by electrophoresis to determine allelic composition at nine enzymatic loci. We observed genetic differentiation between populations of B. cereus and B. thuringiensis. Populations of a given Bacillus species-B. thuringiensis or B. cereus-were genetically more similar to each other than to populations of the other Bacillus species. Hemolytic activity provided further evidence of this genetic divergence, which remained evident even if putative clones were removed from the data set. Our results suggest that the rate of gene flow was higher between strains of the same species, but that exchanges between B. cereus and B. thuringiensis were nonetheless possible. Linkage disequilibrium analysis revealed sufficient recombination for B. cereus populations to be considered panmictic units. In B. thuringiensis, the balance between clonal proliferation and recombination seemed to depend on location. Overall, our data indicate that it is not important for risk assessment purposes to determine whether B. cereus and B. thuringiensis belong to a single or two species. Assessment of the biosafety of pest control based on B. thuringiensis requires evaluation of the extent of genetic exchange between strains in realistic natural conditions.

DESENVOLVIMENTO DE CALOS em EXPLANTES DE CUPUAÇUZEIRO (Theobroma grandiflorum Schum.) em FUNÇÃO da CONCENTRAÇÃO DE AUXINAS E do MEIO LÍQUIDO

Objetivou-se estudar o efeito da concentração de auxina e do meio líquido sobre o desenvolvimento de calos de cupuaçuzeiro. Segmentos de eixos embrionários e cotilédones, obtidos de frutos de cupuaçu dos tipos Mamorana e Redondo, foram cultivados em 4 meios de cultura diferentes: 1) meio MS (50%), suplementado com 2,4-D (1; 2; 4; 8 mg/L); 2) sais N6 (SIGMA) (4 g/L), acrescidos de 2,4-D (0; 2; 4 mg/L) e ANA (0; 3; 5 mg/L); 3) igual ao anterior, suplementado apenas com ANA (3 mg/L); e 4) meio MS, acrescido com ANA (1 mM). Calos com aspecto branco e brilhante foram observados em segmentos de eixos embrionários e cotilédones, cultivados nas menores concentrações de meio 1 (1 e 2 mg/L), enquanto nas maiores concentrações (4 e 8 mg/L) se observou a formação de calos e massa calosa branco-opaca, em eixos embrionários e em segmentos cotiledonares, estas estruturas tornaram-se escuras dentro de oito semanas. Usando o meio 2, um grande número de raízes foram formadas, enquanto o mesmo meio suplementado apenas com ANA (3; 5 mg/L) originou uma massa calosa. A combinação de ANA e 2,4-D, 3 e 2 mg/L, respectivamente, promoveu a formação de calos brancos e raízes. A transferência das culturas para meio líquido, sem regulador de crescimento, promoveu aumento de tamanho dos explantes e escurecimento dos mesmos. O cultivo desses explantes no meio 3 resultou no aparecimento de calos amarelos, com aspecto friável, que permaneceram com a mesma aparência no meio 4.

Draft Genome Sequence of the Nitrogen-Fixing Symbiotic Bacterium Bradyrhizobium elkanii 587

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

Does Immunotherapy Protect Equines from Reinfection by the Oomycete Pythium insidiosum?

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Sodium levels for Japanese quail at initial phases

The aim of this research was to determine the nutritional requirements of sodium for Japanese quail (Coturnix coturnix Japonica) during the periods of 1 to 21 d and 22 to 40 d of age, as well as to evaluate the residual effect on egg production and densitometry bone traits from 41 to 63 d. Two experiments were developed. Experiment 1: 360 Japanese quail were used, from 1 to 21 d of age. Treatments consisted of 5 sodium levels (0.06, 0.12, 0.18, 0.24, and 0.30%). Experiment 2: 240 Japanese quail were used, from 22 to 40 d. Treatments consisted of 5 sodium levels (0.04, 0.12, 0.20, 0.28, and 0.36%). In both experiments, weight gain, final weight, and feed conversion presented a quadratic trend, whereas water intake presented a linear trend. Treatments did not affect the densitometry of bone traits, although they presented a quadratic influence on tibia ash, calcium, and calcium: phosphorus ratio. Therefore, the nutritional requirement of sodium for Japanese quail from 1 to 21 d and from 22 to 40 d is 0.222% and 0.253%, respectively.

Development and application of a Saccharomyces cerevisiae-expressed nucleocapsid protein-based enzyme-linked immunosorbent assay for detection of antibodies against infectious bronchitis virus

A Saccharomyces cerevisiae-expressed nucleocapsid (N) polypeptide of the M41 strain of infectious bronchitis virus (IBV) was used as antigen in a recombinant yeast-expressed N protein-based enzyme-linked immunosorbent assay (Y-N-ELISA). The Y-N-ELISA was rapid, sensitive, and specific for detecting chicken serum antibodies to IBV, and it compared favorably with a commercial ELISA.

Bacterial functional redundancy along a soil reclamation gradient

A strategy to measure bacterial functional redundancy was developed and tested with soils collected along a soil reclamation gradient by determining the richness and diversity of bacterial groups capable of in situ growth on selected carbon substrates. Soil cores were collected from four sites along a transect from the Jamari tin mine site in the Jamari National Forest, Rondonia, RO, Brazil: denuded mine spoil, soil from below the canopy of invading pioneer trees, revegetated soil under new growth on the forest edge, and the forest floor of an adjacent preserved forest. Bacterial population responses were analyzed by amending these soil samples with individual carbon substrates in the presence of bromodeoxyuridine (BrdU), BrdU-labeled DNA was then subjected to a 16S-23S rRNA intergenic analysis to depict the actively growing bacteria from each site, the number and diversity of bacterial groups responding to four carbon substrates (L-serine, L-threonine, sodium citrate, and or-lactose hydrate) increased along the reclamation-vegetation gradient such that the preserved forest soil samples contained the highest functional redundancy for each substrate. These data suggest that bacterial functional redundancy increases in relation to the regrowth of plant communities and may therefore represent an important aspect of the restoration of soil biological functionality to reclaimed mine spoils. They also suggest that bacterial functional redundancy may be a useful indicator of soil quality and ecosystem functioning.

Exigências de lisina digestível para aves de corte da linhagem ISA Label criadas em semiconfinamento

Foram realizados três experimentos para determinar as exigências de lisina digestível para aves da linhagem ISA Label, de ambos os sexos, criadas em semiconfinamento durante as fases: inicial (1 a 28 dias), de crescimento (28 a 56 dias) e final (56 a 84 dias). em cada experimento, foram utilizadas 480 aves, alojadas em 24 piquetes, cada um contendo abrigo coberto de 3,13m² e área de pastejo de 72,87m². O delineamento experimental utilizado foi o inteiramente ao acaso, em esquema fatorial 4x2 (níveis de lisina e sexo) com três repetições de 20 aves cada. Os níveis de lisina digestível avaliados foram: 0,850; 0,970; 1,090 e 1,210% na fase inicial; 0,750; 0,870; 0,990 e 1,110% na fase de crescimento e 0,640; 0,760; 0,880 e 1,000% na fase final. Foram mensuradas as variáveis de desempenho, característica de carcaça, deposição de proteína e gordura corporal, peso e teor de proteína das penas. Com base nos resultados de desempenho, recomendam-se 1, 041; 1,006 e 0,760% de lisina digestível em rações para aves ISA Label nas fases inicial, de crescimento e final, respectivamente.

Influência da altura das garras dos pneus de um trator em área de plantio direto

This work was carried out to evaluate the performance of a farm tractor fitted with two sets of tires with high lugs and another set of tires with tallow lugs in straw without tillage (corn straw). The travel speeds used were approximately 4, 5, 6 and 7 km h(-1) and a constant pulling force of 25 kN was fixed. Tractor traction, forward speed, slip and consumption of fuel were measured and drawbar power, the ratio between the consumption and power and traction coefficient were calculated. It was observed that the tractor performance was similar to high and low lug tire conditions, in an area covered with corn straw.

Effect of Usnic Acid on Candida orthopsilosis and C. parapsilosis

The activity of usnic acid against Candida orthopsilosis and Candida parapsilosis on planktonic and biofilm conditions was investigated by using a broth microdilution and microplate methods. Potent in vitro activities against different Candida species were obtained. The metabolic activity of sessile cells of C. parapsilosis complex was reduced by 80% at four times the 80% inhibitory concentration. The in vitro studies support further efforts to determine whether usnic acid can be used clinically to cure patients with Candida infections.

Altered Ex Vivo Expression of Caspase 8, Caspase 9, and Bcl-2 Is Associated with T-Cell Hyporeactivity in Patients with Paracoccidioidomycosis

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