Lateral hypothalamus lesions influences water and salt intake, and sodium and urine excretion, and arterial blood pressure induced by L-NAME and FK 409 injections into median preoptic nucleus in conscious rats

Male Holtzman rats weighting 200-250 g were anesthetized with zoletil 50 mg/Kg (tiletamine chloridrate 125,0 mg and zolazepan chloridrate 125,0 mg) into quadriceps muscle and submitted an electrolytic lesion of the lateral hypothalamus (LH) and a stainless steel cannula was implanted into their median preoptic nucleus (MnPO). We investigated the effects of the injection into the (MnPO) of FK 409 (20 mug/0.5 mul), a nitric oxide (NO) donor, and N-W-nitro-L-arginine methyl ester (L-NAME) 40 mug/0.5 mul, a nitric oxide synthase inhibitor (NOSI), on the water and sodium appetite and the natriuretic, diuretic and cardiovascular effects induced by injection of L-NAME and FK 409 injected into MnPO in rats with LH lesions. Controls were injected with a similar volume of 0.15 M NaCl. L-NAME injected into MnPO produced an increase in water and sodium intake and in sodium and urine excretion and increase de mean arterial pressure (MAP). FK 409 injected into MnPO did not produce any change in the hydro electrolytic and cardiovascular parameters in LH-sham and lesioned rats. FK 409 injected before L-NAME attenuated its effects. These data show that electrolytic lesion of the LH reduces fluid and sodium intake as well as sodium and urine excretion, and the pressor effect induced by L-NAME. LH involvement with NO of the MnPO excitatory and inhibitory mechanisms related to water and sodium intake, sodium excretion and cardiovascular control is suggested. (C) 2004 Elsevier B.V. All rights reserved.

Isolation and characterization of Flavobacterium columnare (Bernardet et al. 2002) from four tropical fish species in Brazil

Flavobacterium columnare é o agente etiológico da columnariose em peixes de água doce, ocasionando enfermidade na pele e nas brânquias, provocando freqüentemente um grande número de mortalidade. O objetivo deste estudo foi o isolamento e a caracterização de Flavobacterium columnare em peixes tropicais no Brasil. Piracanjuba (Brycon orbignyanus), pacu (Piaractus mesopotamicus), tambaqui (Colossoma macropomum) e cascudo (Hypostomus plecostomus) foram examinados externamente com relação a sinais característicos de columnariose, como manchas acinzentadas na cabeça, região dorsal e pedúnculo caudal dos peixes. A amostragem compreendeu a coleta de 50 exemplares de peixes, representando as quatro diferentes espécies escolhidas para este estudo. Amostras para o isolamento foram obtidas através de raspado com swab estéril das lesões e do rim dos peixes clinicamente diagnosticados como acometidos por columnarios e imediatamente semeados em meios de culturas artificiais (líquido e sólido) próprios para o estudo de Flavobacterium segundo Carlson e Pacha (1968). No meio líquido, houve o desenvolvimento de microrganismos que observados em gota pendente apresentaram a forma de bacilos finos, longos, móveis por deslizamento. Através da coloração de Gram, apresentaram morfologia de bacilos finos, Gram negativos, agrupados em colunas. em meio sólido, as colônias eram pequenas, cinza-amareladas, com borda em forma de raiz. No total, foram obtidos quatro isolamentos: 01 cepa de Brycon orbignyanus; 01 cepa de Piaractus mesopotamicus; 01 cepa de Colossoma macropomum; e 01 cepa de Hypostomus plecostomus. A caracterização bioquímica das amostras, como absorção do vermelho Congo, produção de flexirrubina, produção de H2S e redução do nitrato, sugere que os isolamentos poderiam ser classificados como Flavobacterium columnare.

Expression of the sigma35 and cry2AB genes involved in Bacillus thuringiensis virulence

Muitos genes estão envolvidos nos mecanismos de esporulação da bactéria Bacillus thuringiensis. A regulação e expressão desses genes resultam em uma produção massiva da proteína Cry, responsável pela morte das larvas de muitos insetos. Neste trabalho monitorou-se a expressão de genes de Bacillus thuringiensis, ao longo de três fases de seu desenvolvimento. Foram construídos macroarrays de DNA dos genes selecionados, cujas seqüências estão disponibilizadas no GenBank. Estes genes foram hibridizados com cDNAs obtidos de B. thuringiensis kurstaki HD-1. As sondas de cDNA foram sintetizadas a partir da transcrição reversa do RNA da bactéria, extraído durante as fases de crescimento logarítmico, estacionária e esporulativa, marcadas com 33PadCTP. A expressão diferencial encontrada foi significativa para dois genes de B.thuringiensis, um relacionado aos fatores sigma (sigma35) e outro ao gene cry (cry2Ab). Detectaram-se diferenças entre as médias de expressão do fator sigma e do gene cry2Ab. Os valores máximos de expressão diferencial foram obtidos para o gene codificador do fator sigma35 na fase log e na fase esporulativa. Na análise de médias observou-se expressão do gene cry2Ab apenas na fase log; no entanto, de forma bem mais baixa quando comparado com a expressão de sigma35, nas três fases.

The feeding preferences of Spodoptera frugiperda (J. E. SMITH) (Lepidoptera: Noctuidae) on cotton plant varieties

Avaliou-se atratividade e não-preferência alimentar de lagartas recém-eclodidas de Spodoptera frugiperda por partes de plantas e plantas de variedades de algodoeiro. Testes foram realizados a 27 ± 1ºC, UR de 70% ± 10% e fotofase de 14h. Folhas, brácteas, botões florais e cascas de maçãs da variedade BRS Itamarati-90 e folhas de Fibermax-966, Fibermax-977, DeltaOpal, DeltaPenta, BRS Acala-90, Coodetec-408, Coodetec-409, Coodetec-410, BRS-Cedro, BRS-Ipê, BRS-Aroeira, IPR-96, IPR-120, BRS-Araçá, IAC-24 e BRS Itamarati-90 foram utilizadas nos testes de atratividade e não-preferência para alimentação, com e sem chance de escolha. Utilizaram-se 20 lagartas de S. frugiperda por placa de Petri (sistema de arena) por teste, com 10 repetições. Contaram-se lagartas para avaliar atratividade por 60 min e não-preferência para alimentação por 24 h. Folha foi mais atrativa e preferida para alimentação por lagartas de S. frugiperda. em livre escolha, Coodetec-410 foi mais atrativa e BRS Acala-90, Fibermax-966 e DeltaPenta, as de menor atratividade à S. frugiperda; BRS-Araçá, mais preferida para alimentação e BRS-Cedro, BRS Itamarati 90, DeltaPenta, Coodetec-408 e BRS-Aroeira, menos preferidas. Considera-se 46 min., tempo mais adequado para avaliar atratividade de algodoeiro a lagartas de S. frugiperda.

Processamento mínimo em goiabas 'Paluma' e 'Pedro Sato': 2. Avaliação química, sensorial e microbiológica

Foram utilizados frutos de goiabeiras das cultivares Pedro Sato e Paluma, provenientes de pomar comercial, no estádio de maturação de vez, correspondente à coloração verde-mate e considerados ótimo para o consumo. Os frutos foram inicialmente imersos em solução de hipoclorito de sódio (150mg de cloro.L-1) por 5 minutos, para desinfecção superficial. Pessoas treinadas, utilizando proteção adequada e equipamentos desinfetados descascaram os frutos, cortaram-nos longitudinalmente ao meio e eliminaram a polpa com as sementes, em ambiente a 12°C. Após enxágüe com água clorada (20mg de cloro.L-1) foram embalados em contentores de tereftalato de polietileno (PET) com tampa. Estas unidades foram armazenadas a 3°C por 10 dias. Foram realizadas análises microbiológicas ao longo do período. Determinaram-se quimicamente os conteúdos de lignina, ácido ascórbico, acidez total titulável, sólidos solúveis totais e porcentagem de solubilização das pectinas, bem como as variáveis sensoriais de textura, sabor e preferência. A textura tornou-se mais frágil e os conteúdos de ácido ascórbico se reduziram, ao longo do período de armazenamento, nos produtos de ambas as cultivares. Durante este período houve aumento no conteúdo de lignina e manutenção dos conteúdos de sólidos solúveis totais (SST), acidez total titulável (ATT) e da relação SST/ATT. O produto da cultivar Pedro Sato apresentou menor perda de textura que o da 'Paluma', sendo considerado pelos provadores como o mais saboroso e, portanto, o mais preferido. Devido aos cuidados higiênicos tomados durante o processamento o produto apresentou baixa contagem microbiana (< 10³ UFC.g-1), em todas as avaliações efetuadas.

Uso de agentes microbianos e químico para o controle de Meloidogyne incognita em soja

Nematoides de galhas constituem importante grupo de patógenos da cultura da soja e o manejo integrado é uma das principais medidas de controle que visam à redução de perdas econômicas. Neste trabalho foi avaliada a eficácia dos fungos Paecilomyces lilacinus (Thom.) Samsom e Pochonia chlamydosporia (Goddard) Zare & Gams (sinonímia Verticillium chlamydosporium), de um produto comercial à base de Bacillus sp. (Nemix) e do nematicida químico Aldicarb no controle de Meloidogyne incognita em soja, variedade M-SOY 6101. O experimento foi realizado em casa-de-vegetação no delineamento experimental de blocos casualizados com nove tratamentos (três produtos biológicos usados no tratamento de sementes com ou sem a aplicação em pós-emergência, Aldicarb aplicado apenas em pós-emergência e duas testemunhas) e quatro repetições. Aldicarb reduziu o número de ovos e de juvenis do nematoide. P. lilacinus foi o mais atuante dos agentes biológicos, favorecendo a manutenção da quantidade de matéria seca da raiz de soja e reduzindo o número de ovos. O produto Nemix e P. chlamydosporia somente tiveram ação efetiva na redução do número de ovos do nematoide. Com base nos resultados, foi possível concluir que o agente químico e os agentes biológicos avaliados neste trabalho tiveram moderada atividade no controle de M. incognita em soja.

Transcription profiling of signal transduction-related genes in sugarcane tissues

A collection of 237,954 sugarcane ESTs was examined in search of signal transduction genes. Over 3,500 components involved in several aspects of signal transduction, transcription, development, cell cycle, stress responses and pathogen interaction were compiled into the Sugarcane Signal Transduction (SUCAST) Catalogue. Sequence comparisons and protein domain analysis revealed 477 receptors, 510 protein kinases, 107 protein phosphatases, 75 small GTPases, 17 G-proteins, 114 calcium and inositol metabolism proteins, and over 600 transcription factors. The elements were distributed into 29 main categories subdivided into 409 sub-categories. Genes with no matches in the public databases and of unknown function were also catalogued. A cDNA microarray was constructed to profile individual variation of plants cultivated in the field and transcript abundance in six plant organs (flowers, roots, leaves, lateral buds, and 1(st) and 4(th) internodes). From 1280 distinct elements analyzed, 217 (17%) presented differential expression in two biological samples of at least one of the tissues tested. A total of 153 genes (12%) presented highly similar expression levels in all tissues. A virtual profile matrix was constructed and the expression profiles were validated by real-time PCR. The expression data presented can aid in assigning function for the sugarcane genes and be useful for promoter characterization of this and other economically important grasses.

Effects of different soil tillage systems and coverages on soybean crop in the Botucatu Region in Brazil

Nowadays, agricultural practices should combine high yields with a sustainable use of resources. Different tillage practices and crop covers, if combined, may help to achieve both objectives. In this work, several traits of a soybean (Glycine max L. Merr) cultivar were studied under different conditions of tillage and previous soil coverages. The experiment was installed at Lageado Research Station, Botucatu county, SP, Brazil, on a Paleudult. It consisted of nine treatments (combining three systems of soil tillage and three cover crops) and 4 replicates, yielding 36 plots of a randomized block experimental design. The soil tillage systems considered were: (i) conventional tillage with two heavy harrowing and a levelling harrowing; (ii) chiseling, and (iii) no-tillage with chemical drying of vegetation. The three cover crops used were: black oat, sorghum and spontaneous vegetation. Analyzed variables were: plant height, initial and final plant densities, height of first pod insertion, weight of a thousand grains, number of pods per plant, number of grains per pod, and crop yield. No significant differences were observed for most of the analyzed variables; however, conventional tillage produced significantly heavier grains and a higher number of pods per plant. The selected covers were considered an excellent coverage prior to planting soybean in a crop rotation. The three tillage systems can be used for deployment of culture without compromising the development of soybean.

Modelagem de distorções entre sad 69 e sirgas 2000 pelo método de shepard e grades regulares

Brazil follows the tendency of some countries to update and/or review their fundamental geodetic network. The adoption of geocentric referentials like SIRGAS 2000, the new official reference system of the Geodetic Brazilian System has been an advance. Changes in referential implies in coordinates changes on the network stations as well as the network geometry. To make use of analogical and digital products which are already known in the old referentials are necessary approaches to the coordinate conversion, which minimize the distortions between the used reference frames. This paper presents a distortion modeling approach between reference frames, based on distortion grid generation by using the Shepard's method. To analyze the approach some experiments were performed with the generation of a 1 degrees x1 degrees distortion grid to model the distortions between SAD 69 (1996) and SIRGAS (2000) frames. The results in the test stations were promising, with an average reduction of 50% in the RMS coordinates after the distortions modeling.

Polymorphisms of Lewis and Secretor genes are related to breast cancer and metastasis in axillary lymph nodes

ABH and Lewis antigen expression has been associated with cancer development and prognosis, tumor differentiation, and metastasis. Considering that invasive ductal breast carcinoma (IDC) presents multiple molecular alterations, the aim of the present study was to determine whether the polymorphism of ABO, Lewis, and Secretor genes, as well as ABO phenotyping, could be associated with tumor differentiation and lymph nodes metastasis. Seventy-six women with IDC and 78 healthy female blood donors were submitted to ABO phenotyping/genotyping and Lewis and Secretor genotyping. Phenotyping was performed by hemagglutination and genotyping by the polymerase chain reaction with sequence-specific primers. ABO, Lewis, and Secretor genes were classified by individual single nucleotide polymorphism at sites 59, 1067, 202, and 314 of the Lewis gene, 428 of the Secretor gene, and 261 (O1 allele), 526 (O2 and B allele), and 703 (B allele). No association was found between breast cancer and ABO antigen expression (P = 0.9323) or genotype (P = 0.9356). Lewis-negative genotype was associated with IDC (P = 0.0126) but not with anatomoclinical parameters. Nonsecretor genotype was associated with axillary lymph node metastasis (P = 0.0149). In conclusion, Lewis and Secretor genotyping could be useful to predict respectively breast cancer susceptibility and axillary lymph nodes metastasis.

Phenotypic and genotypic characterization of Rhodococcus equi isolated from sputum

Introduction: Rhodococcus equi is an opportunistic pathogen, causing rhodococcosis, a condition that can be confused with tuberculosis. Often, without identifying M. tuberculosis, physicians initiate empiric treatment for tuberculosis. R. equi and M. tuberculosis have different susceptibility to drugs. Identification of R. equi is based on a variety of phenotypic, chromatographic, and genotypic characteristics.Objective: This study aimed to characterize bacterial isolates from sputum samples suggestive of R. equi.Methods: The phenotypic identification included biochemical assays; thin-layer chromatography (TLC) and polymerase chain reaction (PCR) were used for genotypic identification.Results: Among 78 Gram-positive and partially acid-fast bacilli isolated from the sputum of tuberculosis-suspected patients, 51 were phenotypically and genotypically characterized as R. equi based on literature data. Mycolic acid analysis showed that all suspected R. equi had compounds with a retention factor (R-f) between 0.4-0.5. Genotypic characterization indicated the presence of the choE gene 959 bp fragments in 51 isolates CAMP test positive. Twenty-two CAMP test negative isolates were negative for the choE gene. Five isolates presumptively identified as R. equi, CAMP test positive, were choE gene negative, and probably belonged to other bacterial species.Conclusions: The phenotypic and molecular techniques used constitute a good methodological tool to identify R. equi. (C) 2012 Elsevier Editora All rights reserved.

The effect of rifampicin and pyrazinamide on isoniazid pharmacokinetics in rats

Tuberculosis chemotherapy involves combination of the drugs isoniazid (INH), rifampicin (RMP) and pyrazinamide (PYR) for a 6-month period. The present work investigated the influence of RMP and PYR on the pharmacokinetic parameters of INH when groups of rats were pre-treated for 21 days with INH alone or in combination with RMP and/or PYR, in the following amounts per kg body weight: INH 100 mg; INH 100 mg + RMP 100 mg; INH 100 mg + PYR 350 mg; INH 100 mg + PYR 350 mg + RMP 100 mg. It was found that the co-administration of PYR caused an increase in the INH distribution volume (V-d/F), half-life of elimination t(1)/2(beta)) and clearance (Cl-T/F), and a decrease in the area under curve 0 to 24 h (AUC). Co-administration of RMP caused an increase in the Cl-T/F and a decrease in the AUC. The combination INH + PYR + RMP caused an increase in the Cl-T/F and a decrease in the AUC. These significant pharmacokinetic interactions between the tuberculostatic drugs might be related to differences in the therapeutic and toxic effects. Copyright 0( 2007 John Wiley & Sons, Ltd.

Analysis of Factor VIII polymorphic markers as a means for carrier detection in Brazilian families with haemophilia A

Haemophilia A is an X-linked, recessively inherited bleeding disorder of varying severity, which results from the deficiency of procoagulant factor VIII f(8). Linkage diagnosis using polymorphic markers in the f8 gene is widely used to detect carriers. The objective of this study was to verify the informativeness of three polymorphic markers in the Brazilian population, to evaluate the usefulness of such markers in carrier detection procedures. Sixty-three unrelated healthy volunteers and 10 haemophilic families were studied. Two microsatellite repeats and one HindIII RFLP markers were used. Carrier and non-carrier status could be determined in 80% of females investigated. Intron 13 markers presented the highest heterozygosity rate (79%) followed by intron 22 (68%) and intron 19 (57%). When all three markers were used together, linkage analysis informativeness increased significantly. We conclude that these markers are suitable for carrier detection in the Brazilian population and we recommend their use in combination to maximize diagnostic efficiency.

Production of Trichophyton mentagrophytes antigens and their characterization in mice

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

Experimental study of the conventional equation to determine a plate's moment of inertia

In this work, we describe an experimental setup in which an electric current is used to determine the angular velocity attained by a plate rotating around a shaft in response to a torque applied for a given period. Based on this information, we show how the moment of inertia of a plate can be determined using a procedure that differs considerably from the ones most commonly used, which generally involve time measurements. Some experimental results are also presented which allow one to determine parameters such as the exponents and constant of the conventional equation of a plate's moment of inertia.