Affinity purification of 61- and 65-kDa rat brain corticotropin-releasing factor receptors and receptor-associated G proteins.

Corticotropin-releasing factor (CRF) has been shown to have a central role in physiological adaptation to stress. It is recognized for stimulating the release of adrenocorticotropin from the anterior pituitary gland, and has more recently been implicated as a regulator of autonomic and immunological responses to stress. Much confusion has surrounded the characterization of CRF receptors, with proteins of varying molecular weights having been identified but never purified and characterized. Recently, two CRF receptors have been cloned from brain and pituitary gland, but evidence from in-situ hybridization studies suggests that further CRF receptor types exist. We therefore developed two techniques which enable the isolation of CRF receptors from whole rat brain. The use of a solid-phase CRF analogue affinity column and elution using a competing ligand resulted in the purification of a single protein of 61 kDa. A second technique was devised which allowed the co-isolation of associated signalling proteins and the identification of CRF bound species following purification. CRF was covalently cross-linked to receptors and the complex purified using antibodies specific for the ligand. This enabled the purification of a CRF receptor of approximately 65 kDa and associated alpha and beta gamma G protein subunits. This study demonstrates the successful isolation of CRF receptors which are of different molecular weights to those previously observed from affinity cross-linking studies or predicted from cloned genes. In addition, we confirm the involvement of G proteins in CRF stimulated cell signalling by demonstrating their association with purified CRF receptor.

Updated radiative forcing estimates of 65 halocarbons and nonmethane hydrocarbons

The direct radiative forcing of 65 chlorofluorocarbons, hydrochlorofluorocarbons, hydrofluorocarbons, hydrofluoroethers, halons, iodoalkanes, chloroalkanes, bromoalkanes, perfluorocarbons and nonmethane hydrocarbons has been evaluated using a consistent set of infrared absorption cross sections. For the radiative transfer models, both line-by-line and random band model approaches were employed for each gas. The line-by-line model was first validated against measurements taken by the Airborne Research Interferometer Evaluation System (ARIES) of the U.K. Meteorological Office; the computed spectrally integrated radiance of agreed to within 2% with experimental measurements. Three model atmospheres, derived from a three-dimensional climatology, were used in the radiative forcing calculations to more accurately represent hemispheric differences in water vapor, ozone concentrations, and cloud cover. Instantaneous, clear-sky radiative forcing values calculated by the line-by-line and band models were in close agreement. The band model values were subsequently modified to ensure exact agreement with the line-by-line model values. Calibrated band model radiative forcing values, for atmospheric profiles with clouds and using stratospheric adjustment, are reported and compared with previous literature values. Fourteen of the 65 molecules have forcings that differ by more than 15% from those in the World Meteorological Organization [1999] compilation. Eleven of the molecules have not been reported previously. The 65-molecule data set reported here is the most comprehensive and consistent database yet available to evaluate the relative impact of halocarbons and hydrocarbons on climate change.

(65)Zn(2+) transport by isolated gill epithelial cells of the American lobster, Homarus americanus

Gills are the first site of impact by metal ions in contaminated waters. Work on whole gill cells and metal uptake has not been reported before in crustaceans. In this study, gill filaments of the American lobster, Homarus americanus, were dissociated in physiological saline and separated into several cell types on a 30, 40, 50, and 80% sucrose gradient. Cells from each sucrose solution were separately resuspended in physiological saline and incubated in (65)Zn(2+) in order to assess the nature of metal uptake by each cell type. Characteristics of zinc accumulation by each kind of cell were investigated in the presence and absence of 10 mM calcium, variable NaCl concentrations and pH values, and 100 mu M verapamil, nifedipine, and the calcium ionophore A23187. (65)Zn(2+) influxes were hyperbolic functions of zinc concentration (1-1,000 mu M) and followed Michaelis-Menten kinetics. Calcium reduced both apparent zinc binding affinity (K (m)) and maximal transport velocity (J (max)) for 30% sucrose cells, but doubled the apparent maximal transport velocity for 80% sucrose cells. Results suggest that calcium, sodium, and protons enter gill epithelial cells by an endogenous broad-specificity cation channel and trans-stimulate metal uptake by a plasma membrane carrier system. Differences in zinc transport observed between gill epithelial cell types appear related to apparent affinity differences of the transporters in each kind of cell. Low affinity cells from 30% sucrose were inhibited by calcium, while high affinity cells from 80% sucrose were stimulated. (65)Zn(2+) transport was also studied by isolated, intact, gill filament tips. These intact gill fragments generally displayed the same transport properties as did cells from 80% sucrose and provided support for metal uptake processes being an apical phenomenon. A working model for zinc transport by lobster gill cells is presented.

Temperature Dependence of the Intergranular Critical Current Density in Uniaxially Pressed Bi(1.65)Pb(0.35)Sr(2)Ca(2)Cu(3)O(10+delta) Samples

We have studied the normal and superconducting transport properties of Bi(1.65)Pb(0.35)Sr(2)Ca(2)Cu(3)O(10+delta) (Bi-2223) ceramic samples. Four samples, from the same batch, were prepared by the solid-state reaction method and pressed uniaxially at different compacting pressures, ranging from 90 to 250 MPa before the last heat treatment. From the temperature dependence of the electrical resistivity, combined with current conduction models for cuprates, we were able to separate contributions arising from both the grain misalignment and microstructural defects. The behavior of the critical current density as a function of temperature at zero applied magnetic field, J (c) (T), was fitted to the relationship J (c) (T)ae(1-T/T (c) ) (n) , with na parts per thousand 2 in all samples. We have also investigated the behavior of the product J (c) rho (sr) , where rho (sr) is the specific resistance of the grain-boundary. The results were interpreted by considering the relation between these parameters and the grain-boundary angle, theta, with increasing the uniaxial compacting pressure. We have found that the above type of mechanical deformation improves the alignment of the grains. Consequently the samples exhibit an enhance in the intergranular properties, resulting in a decrease of the specific resistance of the grain-boundary and an increase in the critical current density.

Avaliação de processo do Programa de Regionalização do Turismo em 65 municípios turísticos brasileiros

Este estudo apresenta uma pesquisa realizada em 65 municípios turísticos brasileiros, no período de dezembro de 2007 a fevereiro de 2008, cujo objetivo é avaliar a implementação do Programa de Regionalização do Turismo nesses municípios. Este programa é o eixo principal das políticas públicas do turismo brasileiro, lançado pelo Ministério do Turismo em 2004. Para compreensão dessa política pública do turismo, realizou-se uma pesquisa documental nos arquivos do Ministério do Turismo, neste descrevem-se o Plano Nacional de Turismo (2003-2007) e o Programa de Regionalização do Turismo; também se realizou uma revisão de literatura sobre os princípios e conceitos em que se alicerça o programa: descentralização participativa, integração, sustentabilidade e a elaboração de uma matriz para avaliação de processo utilizada neste trabalho. Faziam parte da pesquisa as 27 capitais estaduais, o Distrito Federal e mais 37 municípios localizados em consolidados destinos turísticos (Floresta Amazônica, Pantanal Mato-grossense, Serras Gaúchas, Cidades Históricas de Minas, Litoral do Nordeste e outros). Por meio da pesquisa de campo e observação sistemática in loco, 23 pesquisadores coletaram informações dos gestores de turismo locais, utilizando-se de formulários fechados. Estes formulários forma elaborados tendo em vista os objetivos dos nove módulos operacionais previstos no Programa de Regionalização do Turismo e seus indicadores de resultados previamente determinados. As respostas, depois da tabuladas e calculadas suas frequências, foram transformadas em gráficos de colunas para fornecer uma visão clara da atual situação do programa em relação à sua implementação nos municípios. Analisando os resultados, obteve-se que, dos nove módulos do programa, quatro foram implementados com eficácia restrita nos municípios, necessitando de ajustes em suas ações operacionais, por parte dos municípios; outros quatro módulos alcançaram resultados mais modestos quanto à sua implementação, demandando melhor acompanhamento e correções por conta dos gestores de turismo; e um módulo teve resultado ineficaz, pois foi implementado em apenas sete municípios, este sim, merecendo maior atenção na sua estruturação, nos seus objetivos, competências delegadas e estratégias. Confrontando esses resultados com a revisão teórica aqui levantada, verificou-se que o processo descentralizador aflorou a fragilidade dos municípios que não cumprem com suas atribuições previstas no programa; evidenciou-se uma fraca integração entre municípios e entre setores público/privado, no sentido de formarem ¿redes¿ de relacionamento e mostrou que o principal programa público de turismo do Brasil está carente de monitoramento e avaliação.

Condições de saúde bucal e autopercepção de saúde bucal da população do grupo etário de 65-74 anos da região da serra/Rio Grande do Sul (RS)

Este estudo tem como objetivo compreender a autopercepção das condições de saúde bucal do grupo etário de 65-74 anos da Região da Serra/RS. Utilizou dados do SBBrasil, coletados pela SES/RS. A população final da amostra foi constituída de 618 idosos, sendo 57% de mulheres. Esta coleta foi realizada através de um questionário com questões fechadas sobre dados sócio-demográficos e questões de autopercepção em saúde bucal, bem como de um exame bucal. A análise dos dados foi feita através da regressão logística multinomial. O exame clínico revelou que quase a metade dos idosos está desdentada e classificou sua saúde bucal como boa ou ótima. Em relação a presença de dor, 28,8% dos indivíduos relataram que sentiram dor nos seis meses que antecederam a entrevista. A dor permaneceu estatisticamente associada à classificação da saúde bucal (OR= 2,3; IC95%: 1,24-4,44) e da mastigação (OR=1,9; IC%95: 1,07-3,24). A necessidade de prótese total permaneceu associada, após o ajuste, com a classificação da aparência dos dentes e gengiva (OR=0,3; IC95%: 0,11-0,78), da mastigação (OR=0,2; IC95%: 0,09- 0,46) e da autopercepção da influência da saúde bucal nos relacionamentos (OR=3,4; IC95%: 1,47-7,75). A renda pessoal manteve associação, após o ajuste, com a classificação da fala (OR=4,5; IC95%: 1,34- 15,12). A escolaridade, após o ajuste, manteve associação com a autopercepção da influência da saúde bucal nos relacionamentos (OR=1,9; IC95%: 1,06-3,43). Conclui-se que a dor e a necessidade de prótese total têm forte relação com as questões de autopercepção em saúde bucal.

Genomic Strategy Identifies a Missense Mutation in WD-Repeat Domain 65 (WDR65) in an Individual With Van der Woude Syndrome

Genetic variation in the transcription factor interferon regulatory factor 6 (IRF6) causes and contributes risk for oral clefting disorders. We hypothesized that genes regulated by IRF6 are also involved in oral clefting disorders. We used five criteria to identify potential IRF6 target genes; differential gene expression in skin taken from wild-type and Irf6-deficient murine embryos, localization to the Van der Woude syndrome 2 (VWS2) locus at 1p36-1p32, overlapping expression with Irf6, presence of a conserved predicted-binding site in the promoter region, and a mutant murine phenotype that was similar to the Irf6 mutant mouse. Previously, we observed altered expression for 573 genes; 13 were located in the murine region syntenic to the VWS2 locus. Two of these genes, Wdr65 and Stratifin, met 4 of 5 criteria. Wdr65 was a novel gene that encoded a predicted protein of 1,250 amino acids with two WD domains. As potential targets for Irf6 regulation, we hypothesized that disease-causing mutations will be found in WDR65 and Stratifin in individuals with VWS or VWS-like syndromes. We identified a potentially etiologic missense mutation in WDR65 in a person with VWS who does not have an exonic mutation in IRF6. The expression and mutation data were consistent with the hypothesis that WDR65 was a novel gene involved in oral clefting. (C) 2011 Wiley-Liss, Inc.

Structural and magnetic characterization of MnxZn1-xFe2O4 (x=0.2; 0.35; 0.65; 0.8; 1.0) ferrites obtained by the citrate precursor method

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Synthesis and characterization of PLZT (9/65/35) by the Pechini method and partial oxalate

ZrTiO4 (ZT), obtained by the Pechini method, was used as precursor for obtaining PLZT (lead lanthanum zirconium titanate). An aqueous solution of oxalic acid was prepared with particles of ZT, Pb(NO3)(2) and La2O3. After the PbC2O4 and La2O3 precipitate on ZT particles, the materials were calcined and X-ray diffraction (XRD) showed the cubic phase of PLZT. This material was sintered, in two steps, and a density of about 8.0 g/cm(3) was obtained. After the second sintering the XRD pattern showed the occurrence of tetragonal and rhombohedral phases. This was caused by a stoichiometric deviation and the material showed a high optical transparency. (C) 1998 Elsevier B.V. B.V. All rights reserved.

Structural characterization of organometallic-derived 9.5/65/35 PLZT ceramics

This study consisted of an investigation of the influence of powder preparation on the phase and chemical compositions and microstructure of 9.5/65/35 PLZT materials sintered in an oxygen atmosphere. The powders with the formula Pb0.905La0.095(Zr0.65Ti0.35)(0.976)O-3+3.5 wt.% PbO were prepared by the polymeric organometallic precursor method (the Pechini method and the partial oxalate procedure). Phase composition was determined by X-ray diffraction of powder and EDS analysis, while grain size was determined based on the micrograph obtained from SEM. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Sintering and characterization of PLZT (9/65/35)

PLZT(9/65/35) obtained by association between the Pechini method (ZT) and partial oxalate (PLZT) was prepared. The stoichiometric phase and monophasic (cubic) PLZT obtained by calcination did not occur after sintering. The sintering process, by using two stages, caused a liquid phase formation due to a PbO excess (5 and 10 wt%). Samples with high density (similar to 8 g/cm(3)) and optical transparency(similar to 12%) were obtained. However, an equilibrium between the excess of PbO of sample/atmosphere PbO leads to a segregated PbO phase on the boundaries of the microstructure. A diffusion of Zr, Ti and La ions from PLZT to PbO phase promoted a stoichiometric deviation of the matrix and modified the optical and dielectric characteristics. (C) 2000 Elsevier B.V. Ltd and Techna S.r.l. All rights reserved.

Synthesis and characterization of 9.5/65/35 PLZT prepared from the polymeric precursors

The 9.5/65/35 PLZT was prepared from the polymeric precursors (the Pechini and partial oxalate process) and by sintering in two stages in an oxygen atmosphere. After thermal treatment at 400 degreesC, the powders were calcinated and sintered at 1200 degreesC with slow heating and cooling rates. The second stage of sintering consisted of hot pressing at the same temperature in oxygen atmosphere. After calcination of PLZT powders obtained by both methods, as well as after sintering of PLZT obtained by Pechini process, the paraelectric cubic phase was formed. After sintering of PLZT obtained by partial oxalate procedure, small tetragonality of crystal structure was observed. After hot pressing PLZT was pseudocubic. SEM microstructural analyses were carried out of the sintering and hot pressed samples and indicated the small grain size less than 2 mum. (C) 2001 Elsevier B.V. B.V. All rights reserved.