A common inhibitory binding site for zinc and odorants at the voltage-gated K+ channel of rat olfactory receptor neurons

This study compared the effects of zinc and odorants on the voltage-gated K+ channel of rat olfactory neurons. Zinc reduced current magnitude, depolarized the voltage activation curve and slowed activation kinetics without affecting inactivation or deactivation kinetics. Zinc inhibition was potentiated by the NO compound, S-nitroso-cysteine. The pH- and diethylpyrocarbonate-dependence of zinc inhibition suggested that zinc acted by binding to histidine residues. Cysteine residues were eliminated as contributing to the zinc-binding site. The odorants, acetophenone and amyl acetate, also reduced current magnitude, depolarized the voltage activation curve and selectively slowed activation kinetics. Furthermore, the diethylpyrocarbonate- and pH-dependence of odorant inhibition implied that the odorants also bind to histidine residues. Zinc inhibitory potency was dramatically diminished in the presence of odorants, implying competition for a common binding site. These observations indicate that the odorants and zinc share a common inhibitory binding site on the external surface of the voltage-gated K+ channel.

Site-Directed Mutagenesis of Dimethyl Sulfoxide Reductase from Rhodobacter capsulatus: Characterization of a Y114 → F Mutant

A system for expressing site-directed mutants of the molybdenum enzyme dimethyl sulfoxide reductase from Rhodobacter capsulatus in the natural host was constructed. This system was used to Generate and express dimethyl sulfoxide reductase with a Y114F mutation. The Y114F mutant had an increased k(cat) and increased K-m toward both dimethyl sulfoxide and trimethylamine N-oxide compared to the native enzyme, and the value of k(cat)/K-m was lower for both substrates in the mutant enzyme. The Y114F mutant, as isolated, was able to oxidize dimethyl sulfide with phenazine ethosulfate as the electron acceptor but with a lower k(cat) than that of the native enzyme. The pH optimum of dimethyl sulfide: acceptor oxidoreductase activity in the Y114F mutant was shown to be shifted by +1 pH unit compared to the native enzyme. The Y114F mutant did not form a pink complex with dimethyl sulfide, which is characteristic of the native enzyme. The mutant enzyme showed a large increase in the K-d for DMS. Direct electrochemistry showed that the Mo(V)/Mo(IV) couple was unaffected by the Y114F mutant, but the midpoint potential of the Mo(VI)/Mo(V) couple was raised by about 50 mV. These data confirm that the Y114 residue plays a critical role in oxidation-reduction processes at the molybdenum active site and in oxygen atom transfer associated with sulfoxide reduction.

Vitamin B12 stalls the 80 S ribosomal complex on the hepatitis C internal ribosome entry site

The effect of cyanocobalamin (CNCbl, vitamin 1312) on hepatitis C virus internal ribosome entry site (HCV IRES)-dependent initiation of translation was studied by ribosomal toeprinting and sucrose gradient centrifugation analysis. These results suggested that CNCbl did not inhibit HCV IRES-dependent translation by a competitive binding mechanism. CNCbl allowed 80 S elongation complex formation on the mRNA, but stalled the initiation at that point, effectively trapping the 80 S ribosomal complexes on the HCV TRES. CNCbl had no effect on cap-dependent mRNA, consistent with the known mRNA specificity of this translational inhibitor. To help elucidate the mechanism, comparative data were collected for the well-characterised translation inhibitors cycloheximide and 5'-guanylyl-imidophosphate, Although CNCbl stalled HCV IRES-dependent translation at approximately the same step in initiation as cycloheximide, the mechanisms of these two inhibitors are distinct. (C) 2002 Elsevier Science Ltd. All rights reserved.

U-Series dating of Liujiang hominid site in Guangxi, Southern China

It has been established that modern humans were living in the Levant and Africa ca. 100 ka ago. Hitherto, this has contrasted with the situation in China where no unequivocal specimens of this species have been securely dated to more than 30 ka. Here we present the results of stratigraphic studies and U-series dating of the Tongtianyan Cave, the discovery site of the Liujiang hominid, which represents one of the few well-preserved fossils of modern Homo sapiens in China. The human fossils are inferred to come from either a refilling breccia or a primarily deposited gravel-bearing sandy clay layer. In the former case, which is better supported, the fossils would date to at least similar to 68 ka, but more likely to similar to 111-139 ka. Alternatively, they would be older than, similar to 153 ka. Both scenarios would make the Liujiang hominid one of the earliest modem humans in East Asia, possibly contemporaneous with the earliest known representatives from the Levant and Africa. Parallel studies on other Chinese localities have provided supporting evidence for the redating of Liujiang, which may have important implications for, the origin of modem humans. (C) 2002 Elsevier Science Ltd. All rights reserved.

Site-directed mutagenesis of the ATM promoter: Consequences for response to proliferation and ionizing radiation

Although ATM, the protein defective in ataxia-telangiectasia (A-T), is activated primarily by radiation, there is also evidence that expression of the protein can be regulated by both radiation and growth factors. Computer analysis of the ATM promoter proximal 700-bp sequence reveals a number of potentially important cis-regulatory sequences. Using nucleotide substitutions to delete putative functional elements in the promoter of ATM, we examined the importance of some of these sites for both the basal and the radiation-induced activity of the promoter. In lymphoblastoid cells, most of the mutations in transcription factor consensus sequences [Sp1(1), Sp1(2), Cre, Ets, Xre, gammaIre(2), a modified AP1 site (Fse), and GCF] reduced basal activity to various extents, whereas others [gammaIre(1), NF1, Myb] left basal activity unaffected. In human skin fibroblasts, results were generally the same, but the basal activity varied up to 8-fold in these and other cell lines. Radiation activated the promoter approximately 2.5-fold in serum-starved lymphoblastoid cells, reaching a maximum by 3 hr, and all mutated elements equally blocked this activation. Reduction in Sp1 and AP1 DNA binding activity by serum starvation was rapidly reversed by exposure of cells to radiation. This reduction was not evident in A-T cells, and the response to radiation was less marked. Data provided for interaction between ATM and Sp1 by protein binding and co-immunoprecipitation could explain the altered regulation of Sp1 in A-T cells. The data described here provide additional evidence that basal and radiation-induced regulation of the ATM promoter is under multifactorial control. (C) 2003 Wiley-Liss, Inc.

Comprasnet: o site da transpar??ncia das compras governamentais

O Site www.comprasnet.gov.br, utilizando recurso da Internet, est?? definindo novas estrat??gias para as Compras Governamentais. Entre as funcionalidades existentes, os usu??rios do governo, fornecedores e a sociedade contam com os seguintes servi??os: Legisla????o: trata dos assuntos legais pertinentes, Publica????es: voltado ??s not??cias de interesse comum entre os usu??rios; Servi??os de Livre Acesso: disponibiliza, entre outros, o SICAFWEB, Preg??o Eletr??nico, Consulta Licita????o; Servi??os por Assinatura: agrega valores para atender necessidades espec??ficas dos usu??rios cadastrados. Exemplo: Consulta Licita????es com acesso a itens e editais, Listas Preferenciais, etc. e, o SIASG que concentra F??rum de Discuss??o, Consultas Gerenciais, Homologa????o de Preg??o Eletr??nico, al??m dos diversos m??dulos que o comp??em

Cultural tourism in Northwest Portugal – the case of the world heritage site of Guimarães

A cidade de Guimarães, no noroeste português, é um espaço imbuído de um forte significado simbólico e cultural. A acreditação pela U.N.E.S.C.O. do seu centro histórico como Património Cultural da Humanidade, em Dezembro de 2001, contribuiu significativamente para aumentar o seu potencial em termos turísticos. Na realidade, desde aquela data, tem-se assistido a um aumento sustentado do número de visitantes. O presente capítulo debruça-se sobre a análise de alguns resultados de um inquérito por questionário aplicado aos residentes do município de Guimarães com o objetivo de avaliar a sua perceção dos benefícios que a atividade turística pode trazer. O inquérito foi implementado entre Janeiro e Março de 2010. Os resultados mostram que aqueles revelam uma perceção favorável dos impactos do turismo e que esta perceção está correlacionada com a idade e o nível de instrução dos inquiridos.

The host-tourist interaction in a world heritage site: the case of Guimarães

Guimarães, in the northwest of Portugal, is a city of strong symbolic and cultural significance and its nomination by UNESCO as world heritage, in 2001, enlarged its tourism potential. In this paper we present a few results of a survey that envisaged capturing the Guimarães residents’ perceptions of tourism impacts and their attitudes towards tourists. Specifically, one analyzes the type of relationship that exists between some socio-demographic groups and the perceived tourism impacts, as well as their socio-characteristics and the existing level of interaction between residents and tourists. The survey was implemented between January and March 2010 to a convenience sample of 540 inhabitants of the municipality of Guimarães resulting in 400 questionnaires with complete data. For this, we made use of various statistical techniques. Using a factorial analysis, we can conclude that the three factors used explain 52.3% of the variance contained in the original variables obtained from the survey. By another side, using a logit model in the analysis and taking as the dependent variable the frequent or very frequent contact with tourists, we found that only the variables referred to perceived positive impacts of tourism, education and the place of residence in urban areas have shown to be statistically significant. We are aware of the multiple ways the issue of residents’ perceptions and attitudes towards tourism can be approached and of the difficulties to get useful policy-oriented insights. This paper is a step in that trail.

Does gender affect visiting a world heritage site?

The city of Guimarães in Portugal is a place of strong symbolic and cultural significance, and the nomination of its historical center as a World Heritage Site in 2001 enhanced its tourism potential. This study presents the results of a survey conducted in 2010 and 2011 to capture the profile and motivations of tourists visiting Guimarães as a cultural tourism destination. The study addressed two main issues: whether males and females have similar or different preferences in choosing the city as their destination, and whether there are gender differences in the perception of the attributes of Guimarães. A better understanding of the gendered nature of the destination is a valuable cue for shaping products and services according to visitors’ preferences. The results suggest that both men and women are aware of the main elements responsible for the city’s World Heritage status. That the destination is a Heritage Site that also offers the opportunity to tour the region has a significant positive effect on male tourists’ choice of Guimarães. Regarding the perceived attributes of the city, results indicate minor gender differences with one exception: Women expressed more apprehension than men regarding the perceived security of the destination.

How does gender affect visiting a world heritage site?: the case study of Guimarães

To date few studies have been undertaken in Portugal dealing with the attitudes, motivations, and profile of tourists who visit World Heritage Sites. Also, few studies have dealt with destination image (e.g., Agapito, Mendes & Valle, 2010; Lopes, 2011). As far as it is known, none have approached the issue of gender differences in the choice of a Portuguese heritage destination. Since cultural tourism destinations need to differentiate themselves from each other, appropriate market segmentation must be based on a deep understanding of the customers’ motivations and preferences. Keeping in mind results from empirical literature (e.g., Silberberg, 1995; Beerli & Martin, 2004; Richards, 2004; Pérez, 2009; Sheng, Shen, & Chen, 2008), gender seems to be a possible approach to market segmentation, whether for Guimarães or for other cultural tourism destinations around the world. Located in the north-western region of Portugal, Guimarães is a city of strong symbolic and cultural significance, and the nomination of its historical centre as a World Heritage Site in 2001 enhanced its tourism potential. This study analyses the possible relation between gender and attitudes and motivations towards a World Heritage Site, such as Guimarães. Additionally, the empirical approach used in the study tries to capture differences in the perceived attributes of the city. Commonalities and distinctions within and between groups of tourists, by focusing on the specific characteristic of gender, were analysed. The study addressed two main questions: first, whether males and females have similar or different preferences in choosing the city as their destination; and, second, whether there are gender differences in the perception of the attributes of Guimarães. A better understanding of the gendered nature of the destination is a valuable cue for shaping products and services according to visitors’ preferences.

Does Gender Affect Visiting a World Heritage Site?

The city of Guimar˜aes in Portugal is a place of strong symbolic and cultural significance, and the nomination of its historical center as a World Heritage Site in 2001 enhanced its tourism potential. This study presents the results of a survey conducted in 2010 and 2011 to capture the profile and motivations of tourists visiting Guimar˜aes as a cultural tourism destination. The study addressed two main issues: whether males and females have similar or different preferences in choosing the city as their destination, and whether there are gender differences in the perception of the attributes of Guimar˜aes. A better understanding of the gendered nature of the destination is a valuable cue for shaping products and services according to visitors’ preferences. The results suggest that both men and women are aware of the main elements responsible for the city’s World Heritage status. That the destination is a Heritage Site that also offers the opportunity to tour the region has a significant positive effect on male tourists’ choice of Guimar˜aes. Regarding the perceived attributes of the city, results indicate minor gender differences with one exception: Women expressed more apprehension than men regarding the perceived security of the destination.