Biossíntese e recentes avanços na produção de celulose bacteriana

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

Análise de polimorfismos dos genes da rota quinureniana em pacientes com meningite bacteriana.

Bacterial meningitis (BM) is still an important infectious disease causing death and disability. Invasive bacterial infections of the central nervous systems (CNS) generate some of the most powerful inflammatory responses known, which contributes to neuronal damage. The DNA microarray technology showed alterations in the kynurenine (KYN) pathway that is induced in BM and other diseases associated with inflammation, leading to brain injury. Our main aim was to search SNPs previously described in the KYN path enzymes to investigate a putative association of this SNPs with imbalanced in this pathway in patients with BM. The patients included in this study were 33 males and 24 females, with ages varying from 02 months to 68 years. SNPs were located inside of the domain conserved in KYNU, IDO, KATI and KATII. Primers were designed for analysis of SNPs already described by PIRA-PCR followed by RFLP. The analysis of KYNU+715G/A SNP found a heterozygous frequency of 0.033. We did not found the variant allele of SNP KYNU+693G/A, KATI+164T/C, KATII+650C/T and IDO+434T/G. Despite of previews studies showing the importance of KYN pathway we did not found one association of these SNPs analyzed with susceptibility or severity of MB in study population.

Associação entre polimorfismos em enzimas de reparo de DNA e ocorrência de meningite bacteriana.

Despite advances in vaccine development and therapy, bacterial meningitis (BM) remains a major cause of death and long-term neurological disabilities. As part of the host inflammatory response to the invading pathogen, factors such as reactive oxygen species are generated, which may damage DNA and trigger the overactivation of DNA repair mechanisms. It is conceivable that the individual susceptibility and outcome of BM may be in part determined by non synonymous polymorphisms that may alter the function of crucial BER DNA repair enzymes as PARP-1, OGG-1 and APE-1. These enzymes, in addition to their important DNA repair function, also perform role of inflammatory regulators. In this work was investigated the non synonymous SNPs APE-1 Asn148Glu, OGG-1 Ser326Cys,PARP-1 Val762Ala, PARP-1 Pro882Leu and PARP-1 Cys908Tyr in patients with bacterial meningitis (BM), chronic meningitis (CM), aseptic meningitis (AM) and not infected (controls). As results we found increased frequency of variant alleles of PARP-1 Val762Ala (P = 0.005) and APE-1 Asn148Glu (P=0.018) in BM patients, APE-1 Asn148Glu in AM patients (P = 0.012) and decrease in the frequency of the variant allele OGG-1 Ser326Cys in patients with CM (P = 0.013), regarding the allelic frequencies in the controls. A major incidence of individuals heterozygous and/ or polymorphic homozygous in BM for PARP-1 Val762Ala (P= 0.0399, OD 4.2, 95% IC 1.213 -14.545) and PARP-1 Val762Ala/ APE-1 Asn148Glu (P = 0.0238, OD 11.111, 95% IC 1.274 - 96.914) was observed related to what was expected in a not infected population. It was also observed a major incidence of combined SNPs in the BM patients compared with the control group (P=0.0281), giving evidences that SNPs can cause some susceptibility to the disease. This combined effect of SNPs seems to regulate the principal cytokines and other factors related to BM inflammatory response and point the importance of DNA repair not only to repair activity when DNA is damaged, but to others essential functions to human organism balance.

Ativação local da rota da quinurenina por moduladores inflamatórios durante a meningite bacteriana.

Activation of the kynurenine (KYN) pathway (KP) by modulators of immune system has been observed during several neurological diseases. Here we assessed the association of chemo-/cytokine levels with the concentration of KP metabolites in cerebrospinal fluid (CSF) and plasma samples from patients with bacterial meningitis (BM). All samples were collected from 42 patients diagnosed with acute bacterial meningitis (ABM), aseptic meningitis, tuberculous meningitis and patients without infection neurological disorders. CSF and plasma concentration of metabolites from the KP was assessed by high pressure liquid chromatography (HPLC) and cytokines and chemokines by Bio-plex 200 suspension array system. Concentrations of the KP metabolites KYN and kynurenic acid (KYNA) were significantly higher in CSF of patients with ABM compared to other groups. Tryptophan (TRP), anthranilic acid (AA), 3-hydroxykynurenine (3HK) and 3-hydroxyanthranilic acid (3HAA) did not show statistical significance, although some of them presented a good accumulation during ABM. The expression of TNF-alpha, IL-6, IL-1beta, IFN-gamma, IL-10, IL-1 receptor antagonist (IL-1Ra), MIP-1alpha, MIP-1beta, MCP-1 and G-CSF was about 100-fold higher in CSF from ABM patients than other infected groups. In all CSF and plasma samples, the concentration of IL-2, IL-12(p70), IL-4, IL-8 and GM-CSF was not significant. ABM still showed significant concentrations of IL-6, IL-10, IL-1Ra and MCP-1 in plasma samples. Based on the comparison of KP metabolites concentrations between plasma and CSF samples we conclude that the activation of the tryptophan pathway upon BM occurs within the brain. This increase in KP metabolites is most due to activation of the KP by molecules as IFN-gamma and TNF-alpha in response to infection.

Método multirresíduo para monitoramento de contaminação ambiental de pesticidas na região de Bauru (SP) usando mel como bio-indicador

The presence of residues of the major groups of pesticides (organohalogen, organophosphorous, pyrethroids and organonitrogen) in representative samples of honey produced in Bauru (state of São Paulo, Brazil) was investigated from 1999 through2004. A multiresidue method was applied to honey samples to determine 48 pesticides with recoveries ranging from 76 to 95%. The limits of detection found were lower than 10 µg/kg for GC-MS-SIM. The results indicated that most pesticides found in the samples belonged to the organohalogen and organonitrogen groups. Residues of malathion were detected in almost all of the samples in high concentration.

Fatores de influência na produção de leite bovino sobre os níveis da contagem bacteriana total

The objective of this study was to evaluate the influence of milking procedures on the levels of total bacterial count (TBC) in bovine milk. In the first study the influences of procedures for hygienic milking, cleaning of milking equipment and milk cooling tanks on the TBC levels were evaluated. Four bulk samples of milk were collected from each tank in eight properties for TBC analysis, employing the flow cytometry method. A questionnaire was applied in each property to assess the current situation of milking procedures on each production system that took part on this research, followed by training of employees in good agricultural practices in the production of milk and monitoring of the TBC measurements. The methodology for analysis of longitudinal data was considered, focusing on random effects models. The results showed that the handling procedures for milking and the cleanliness of the cooling tank contributed to a further reduction in the levels of TBC raw milk cooling tanks. The second study aimed to describe the percentage of the properties that comply with the Normative Instruction Nº 51 (Brazil s IN 51) with regard to total bacterial count (TBC) in bovine milk. The study was conducted from January 2010 to July 2011. Milk samples were collected from the eight properties selected for TBC analysis by the flow cytometry method. Again, on each property a questionnaire was applied to assess the current situation of milking procedures on each production system that took part on this research, followed by training of employees in good agricultural practices in the production of milk and monitoring of the TBC measurements. The methodology of marginal models based on Generalized Estimate Equations (GEEs) was followed in the statistical analysis. The results showed that the handling procedures of the milking and the cleanliness of the cooling tanks contributed to a considerable percentage of the properties that reached the limits of TBC established by IN 51

Efeito da colhedora, velocidade e ponto de coleta na contaminação de sementes de soja por fungos

A pesquisa foi realizada com o objetivo de avaliar a qualidade sanitária de sementes de soja cv. Coodetec 217, colhidas por duas colhedoras operando em velocidades e pontos de coleta distintos. O delineamento foi inteiramente casualizado, no esquema fatorial 2x2x2. Os tratamentos foram constituídos por duas máquinas Massey Ferguson (MF 2003 e MF 2004), duas velocidades (4 e 7 Km.h¹) e dois pontos de coleta (tanque graneleiro e saída do tubo de descarga). A sanidade foi avaliada pelo método do papel de filtro, com quatro repetições de 50 sementes. Houve interação significativa para os fatores colhedora e ponto de coleta para a incidência de Penicillium sp. e Fusarium sp., entre colhedora e velocidade para Aspergillus spp., e, entre velocidade e ponto de coleta para Penicillium sp.. A presença de fungos associados às sementes, principalmente, Cladosporium sp. e Fusarium sp., independe do tipo de colhedora. Uma menor contaminação fúngica é observada nas sementes coletadas na saída do tubo de descarga à 4 Km.h-1. Para Fusarium sp., Aspergillus spp., e Penicillium sp. existe uma dependência dos fatores sobre suas incidências nas sementes da cultivar analisada, sob as condições estudadas.

Revitalização do Rio Golandim (RN/Brasil) após décadas de contaminação por efluentes industriais e domésticos: este rio está recuperado?

Environmental pollution causes the loss of the quality of aquatic resources and also affects the health of human beings. The Golandim River is located in São Gonçalo do Amarante (RN Brazil) and had its water recovered seven years ago by measurements of parameters of the water s quality analyses physicochemistry, microbiological and heavy metals. However, it is not well established if this river is truly recovered, so this study provides a wide knowledge about the quality of these waters. Therefore, this investigation was accomplished by assays of ecotoxicology utilizing Ceriodaphnia dubia and of genotoxicity of the river s water using a biomarker Tradescantia Pallida (Trad-MCN). In set, it carried through a study of environmental perception through questionnaires that approached questions related to the profile of the interviewed one, knowledge on the environment and of the river Golandim with the community that lives in the neighborhoods of the river to diagnosis as they perceive the environment where live and its problems. The assays of the water had been carried out by collecting samples in three different sites of the Golandim River. They were collected between the periods of December 2010 (dry season in the northeast of Brazil) and July 2011 (rainy season in the northeast of). The analysis of the data allowed observing that the majority of the inhabitants are adult and presents a global vision of what it is part of the environment, the majority mentions the pollution of the river as one of the problems of the city, considering it serious. The ecotoxicology assay showed that there was not acute toxicity in all three samples collected. Meanwhile, all these three samples demonstrated significant chronic toxicity. The results from the Trad-MCN assay presented an increase in the frequency of micronucleus in one of the sites analyzed (S3) (p<0.01), in both seasons collected. On the other hand, the sites S1 and S2 did not presented a significant increase of micronucleus using this bioassay. The analyses of chemicals detected an increase in the levels of some metals, in different seasons and samples, which can be associates with some compounds found in urban and industrial areas. On the other hand, the physicochemistries parameters demonstrated that the Golandim River is recouped, when compared with the values presented at the CONAMA s legislation. However, these results indicate the presence of compounds capable of inducing chromosomal mutation in plants. On the other hand, the parameters physicistchemistries demonstrate that the river Golandim is if recouping, since when compares the values observed with the legislation of the CONAMA. All these results point to the fact that the Trad-MCN assay was sensitive and efficient biomarker for chromosomal instability and the C. dubia ecotoxicology assay was as though an efficient biomarker of toxicity of water s quality. The results from Trad-MCN associated with the ecotoxicology demonstrates that these analyses are important for environmental monitoring, once the first bioassay described above indicates alterations at the standards of cells and the other one indicates alterations at the standards of organisms. This study alerts for the necessity to carry out biological assays for the analyses of the water s quality

Avaliação da contaminação da câmara anterior na cirurgia de facoemulsificação com implante de lente intra-ocular

Objetivo: Avaliar a existência de contaminação da câmara anterior durante a facectomia por facoemulsificação com implante de lente intra-ocular. Método: Foi realizado estudo prospectivo, avaliando-se 30 pacientes submetidos a facectomia por facoemulsificação com implante de lente intra-ocular, colhendo-se duas amostras de humor aquoso, uma obtida no início e outra no final da cirurgia. As amostras foram semeadas em meio de cultura para germes aeróbios, anaeróbios e fungos. Resultado: Todas as amostras avaliadas resultaram negativas. Conclusão: A contaminação da câmara anterior na cirurgia de facoemulsificação com implante de lente intra-ocular, usando os cuidados necessários, é infreqüente.

Uso do gpr na caracterização da pluma de contaminação gerada por hidrocarbonetos a partir de um posto de combustível na praia de Búzios/RN

This work is the application of geophysical methods, using the Ground Penetrating Radar (GPR), with the objective of survey in a subsurface plume of contamination caused by a disabled gas station. The gas station is located on the Búzios beach in southern coast of the state to Rio Grande do Norte in an Area of Environmental Protection called Bonfim-Guaraíra. The interest to develop this work was the presence of contaminants (hydrocarbons) in a well located on the desktop, previously used for the abstraction of groundwater for residents living near the site. Were raised 15 geophysical survey lines totaling 775,48 lifting and installed 4 piezometer, to confirm the contamination and prepare a pluviometric map that helped in indicating the direction of local groundwater flow, thus showing the direction of movement of the probably plume of contamination. From the processing of the GPR lines was possible to identify two likely phases of contamination according to the classification proposed by Azambuja et al 2000, which are called phase absorbed and dissolved phase

Liberação intraocular de ofloxacina associada a lente de contato biossintética em ceratite bacteriana experimental em cães

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

Contaminação da pastagem com larvas infectantes de nematoides gastrintestinais após o pastejo de ovelhas resistentes ou susceptíveis à verminose

O grau de contaminação da pastagem por larvas de terceiro estágio (L3) de nematoides gastrintestinais foi avaliado em piquetes pastejados por ovelhas resistentes ou susceptíveis à verminose. O experimento foi realizado de 23 de novembro de 2007 a 22 de julho de 2008. Inicialmente, realizou-se a contagem de ovos por grama de fezes (OPG) de amostras individuais de 130 ovelhas adultas da raça Bergamácia. Dessas, foram selecionadas as nove ovelhas com contagens mais elevadas de OPG (susceptíveis) e as 10 com as menores contagens (resistentes). As ovelhas resistentes apresentaram menor contagem de OPG, maiores valores de volume globular, de proteína plasmática total e de eosinófilos sanguíneos, do que as ovelhas susceptíveis. O peso também foi maior no grupo resistente. Na pastagem, foram identificadas larvas de Haemonchus spp., Trichostrongylus spp. e Oesophagostomum spp. em média, as quantidades de L3 de Haemonchus spp. e de Trichostrongylus spp., na pastagem, foram 2,19 e 2,31 vezes, respectivamente, maiores nos piquetes pastejados pelo grupo susceptível do que nos do grupo resistente. Portanto, os animais susceptíveis devem ser eliminados do rebanho a fim de reduzir a contaminação da pastagem e otimizar a profilaxia das infecções por nematoides gastrintestinais.

Aplicação da resistividade e cargabilidade em estudos de contaminação de sedimentos por derivados de hidrocarbonetos

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

Método da eletrorresistividade aplicado no monitoramento temporal da pluma de contaminação em área de disposição de resíduos sólidos urbanos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Monitoramento da atenuação natural de pluma de contaminação pelo método de radar de penetração no solo (GPR)

O monitoramento da atenuação natural em áreas contaminadas tem se mostrado uma técnica alternativa e de baixo custo para a remediação de áreas contaminadas. A degradação por microorganismos é um dos processos mais importantes na atenuação natural de contaminantes, especialmente compostos de fase líquida não aquosa (NAPL). em muitos casos, a ação efetiva deste processo resulta na geração de ácidos orgânicos, que sob elevadas concentrações ocasionam a dissolução de minerais presentes em subsuperfície onde se encontra a contaminação, com conseqüente liberação de íons. O aumento na quantidade de íons colabora para o aumento da condutividade elétrica do meio. O princípio físico da técnica de Radar de Penetração no Solo (GPR) é a emissão de ondas eletromagnéticas de alta freqüência. A propagação da onda eletromagnética é condicionada à freqüência de sinal emitido e as propriedades elétricas do meio. O aumento da condutividade elétrica do meio resulta na atenuação do sinal e, por conseqüência, na diminuição da profundidade de penetração da onda eletromagnética. Este fator permite o monitoramento de áreas contaminadas sob atenuação natural a partir de análises temporais com o GPR. Este trabalho apresenta um estudo comparativo entre perfis de GPR adquiridos em 1998 e 2003 em uma área contaminada por compostos de fase líquida leve não aquosa (LNAPL), sob atenuação natural. Os resultados indicam um aumento da condutividade elétrica do meio, a partir da atenuação acentuada do sinal GPR observada nas seções de 2003. Este aumento pode estar associado à liberação de íons por dissolução de minerais, pelo ataque de ácidos orgânicos resultantes do processo de biodegradação.