Syntaxin 7 complexes with mouse Vps10p tail interactor 1b, Syntaxin 6, vesicle-associated membrane protein (VAMP)8, and VAMP7 in B16 melanoma cells

Syntaxin 7 is a mammalian target soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) involved in membrane transport between late endosomes and lysosomes. The aim of the present study was to use immunoaffinity techniques to identify proteins that interact with Syntaxin 7. We reasoned that this would be facilitated by the use of cells producing high levels of Syntaxin 7, Screening of a large number of tissues and cell lines revealed that Syntaxin 7 is expressed at very high levels in B16 melanoma cells. Moreover, the expression of Syntaxin 7 increased in these cells as they underwent melanogenesis. From a large scale Syntaxin 7 immunoprecipitation, we have identified six polypeptides using a combination of electrospray mass spectrometry and immunoblotting. These polypeptides corresponded to Syntaxin 7, Syntaxin 6, mouse Vps10p tail interactor 1b (mVti1b), alpha -synaptosome-associated protein (SNAP), vesicle-associated membrane protein (VAMP)8, VAMP7, and the protein phosphatase 1M regulatory subunit. We also observed partial colocalization between Syntaxin 6 and Syntaxin 7, between Syntaxin 6 and mVti1b, but not between Syntaxin 6 and the early endosomal t-SNARE Syntaxin 13. Based on these and data reported previously, we propose that Syntaxin 7/mVti1b/Syntaxin 6 may form discrete SNARE complexes with either VAMP7 or VAMPS to regulate fusion events within the late endosomal pathway and that these events may play a critical role in melanogenesis.

Relationship of high energy expenditure and variation in dietary intake with reporting accuracy on 7 day food records and diet histories in a group of healthy adult volunteers

Objectives: To assess the accuracy of reporting from both a diet history and food record and identify some of the characteristics of more accurate reporters in a group of healthy adult volunteers for an energy balance study. Design: Prospective measurements in free-living people. Setting: Wollongong, Australia. Subjects: Fifteen healthy volunteers (seven male, eight female; aged 22 -59 y; body mass index (BMI) 19 - 33 kg/m(2)) from the local community in the city of Wollongong, Australia. Interventions: Measurement of energy intake via diet history interview and 7 day food records, total energy expenditure by the doubly labelled water technique over 14 days, physical activity by questionnaire, and body fat by dual-energy X-ray absorptiometry. Results: Increased misreporting of energy intake was associated with increased energy expenditure (r = 0.90, P < 0.0001, diet history; r(s)=0.79, P=0.0005, food records) but was not associated with age, sex, BMI or body fat. Range in number of recorded dinner foods correlated positively with energy expenditure (r(s)=0.63, P=0.01) and degree of misreporting (r(s)=0.71, P=0.003, diet history; r(s)=0.63, P=0.01, food records). Variation in energy intake at dinner and over the whole day identified by the food records correlated positively with energy expenditure (r=0.58, P = 0.02) and misreporting on the diet history (r=0.62, P=0.01). Conclusions: Subjects who are highly active or who have variable dietary and exercise behaviour may be less accurate in reporting dietary intake. Our findings indicate that it may be necessary to screen for these characteristics in studies where accuracy of reporting at an individual level is critical. Sponsorship: The study was supported in part by Australian Research Council funds made available through the University of Wollongong.

2,6,,9-trioxabicyclo[3.3.1]nona-3,7-dienes and 2,4,6,8-tetraoxaadamantanes: Novel chiral spacer units in macrocyclic polyethers

The unusual chiral heterocyclic systems, trioxabicyclo[3.3.1]nona-3,7-dienes (bridged bisdioxines), are incorporated as novel spacer molecules into macrocyclic polyether ring systems of various sizes (8, 9 as well as 11-15) by cyclocondensation reaction of the! bisacid chloride 4b or bisesters 6,7 and 10, with several ethylene glycols. The 2:2 macrocycles 12-14 are obtained in approximately 50:50 mixtures of diastereomers. These conclusions are mainly based on HPLC data presented in Table I as well as X-ray analyses of (1R,5R)-8c (space group Pbca, a = 10.163(3) Angstrom, b = 18.999(4) Angstrom, c = 36.187(10) Angstrom, V = 6987(3) Angstrom(3), Z = 8, d(calc) = 1.218 g cm(-3), 6974 reflections, R = 0.0553.), mesolrac-11 (space group P (1) over bar, a = 10.472(5) Angstrom, b = 16.390(5) Angstrom, c = 17.211(5) Angstrom, alpha = 98.69(2)degrees, beta = 93.04(2)degrees, gamma = 98.52(2)degrees, V = 2879.3(18) Angstrom(3), Z = 2, d(calc) = 1.173 g cm(-3), 11,162 reflections, R = 0.0945) and meso-12 (space group P2(1)/c, a = 9.927(2), b = 18.166(3), c = 17.820(3) Angstrom, beta = 96.590(10)degrees, V = 3192.3(10)Angstrom(3), Z = 4, D-c = 1.109 g cm(-3), 3490 reflections, R = 0.0646). The 1:1 macrocycles 8b,c are also formed by intramolecular transesterification of the open-chain bisesters 7b,c and their formation is favored by the use of metal ions as templates. The bridged bisdioxine moieties in 8b and 12 are converted into the corresponding chiral tetra-oxaadamantane spacers to afford macrocycles 16 and 17. Preliminary metal ion complexation studies with selected species (8c, 11-14) were also performed.

Hydrolytic kinetic resolution of terminal mono- and bis-epoxides in the synthesis of insect pheromones: routes to (-)-(R)- and (+)-(S)-10-methyldodecyl acetate, (-)-(R)-10-methyl-2-tridecanone, (-)-(R)-(Z)-undec-6-en-2-ol (Nostrenol), (-)-(1R,7R)-1,7-dime

Hydrolytic kinetic resolution (HKR) of functionalised epoxides using (salen)Co(OAc) complexes provides enantiomerically enriched epoxides and diols, which have been transformed into important insect sex pheromones. In this general approach, (-)-(R)- and (+)-(S)-10-methyldodecyl acetates from the smaller tea tortrix moth were obtained, as was (-)-(R)-10-methyltridecan-2-one from the southern corn rootworm. The (S)-epoxide obtained from undec-1-en-6-yne was transformed to (-)-(R)-(Z)-undec-6-en-2-ol (Nostrenol) from ant-lions. HKR of appropriate bisepoxides was also investigated, and transformations of the resulting bisepoxides and epoxydiols provided (-)-(1R,7R)-1,7-dimethylnonylpropanoate from corn rootworms, (-)-(6R,12R)-6,12-dimethylpentadecan-2-one from the female banded cucumber beetle, and (-)-(2S,11S)-2,11-diacetoxytridecane and (+)-(2S,12S)-2,12-diacetoxytridecane from female pea-midges. (C) 2002 Elsevier Science Ltd. All rights reserved.

Variation in coumarin 7-hydroxylase activity associated with genetic polymorphism of cytochrome P450 2A6 and the body status of iron stores in adult Thai males and females

The relationships between catalytic activity of cytochrome P450 2A6 (CYP2A6), polymorphism of CYP2A6 gene, gender and levels of body iron stores were analysed in a sample group of 202 apparently healthy Thais, aged 1947 years. Eleven individuals were found to have high activity of CYP2A6, judged by the relatively large amounts (11.2-14.6 mg) of 7-hydroyxcoumarin (7-OHC) excreted 3 h following administration of 15 mg of coumarin. Ten individuals, however, did not excrete any 7-OHC. Of these 10, four were found to have no CYP2A6 gene (whole gene deletion; CYP2A6*4 allele). The frequency of the CYP2A6 alleles; *1A, *1B and *4 in the whole sample group was 52, 40 and 8% while the frequency of the CYP2A6 gene types; *1A/* 1A, *1A/* 1B, *1B/* 1B, *1A/* 4, *1BI* 4, *4/* 4 was 29, 41, 16, 7, 5 and 2%. Subjects having CYP2A6* 1A/* 1B gene-type group were found to have higher rates of coumarin 7-hydroxylation compared with those of the CYP2A6* 1B/* 1B and CYP2A6* 1A/* 4 gene types. The inter-individual variability in CYP2A6 catalytic activity was therefore attributed in part to the CYP2A6 genetic polymorphism. Variation in CYP2A6 activity in this sample group was not associated with gender but, interestingly, it did show an inverse association with plasma ferritin; an indicator of body iron stores. Higher rates of coumarin 7-hydroxylation were found in individuals with low body iron stores (plasma ferritin < 20 μg/l) compared with subjects having normal body iron store status. Subjects (n = 16) with iron overload (plasma ferritin > 300 mug/l) also tended to have elevated rates of coumarin 7-hydroxylation. These results suggest an increased CYP2A6 expression in subjects who have excessive body iron stores. Further investigations into the underlying factors that may lead to increased expression of CYP2A6 in association with abnormal body iron stores are currently in progress in our laboratory. Pharmacogenetics 12:241-249 (C) 2002 Lippincott Williams Wilkins.

Decreased coumarin 7-hydroxylase activities and CYP2A6 expression levels in humans caused by genetic polymorphism in CYP2A6 promoter region (CYP2A6*9)

One of seven poor metabolizers of coumarin found in Thai subjects was previously genotyped as heterozygote for the CYP2A6*4 (whole deletion) and CYP2A6*9. Thus, we aimed to investigate the relationship between the genetic polymorphism in the TATA box of the CYP2A6 gene (CYP2A6*9), expression levels of CYP2A6 mRNA and coumarin 7-hydroxylase activities in human livers. Levels of CYP2A6 mRNA were quantified by real-time quantitative reverse transcriptase-polymerase chain reaction. The mean expression levels of CYP2A6 mRNA in individuals with CYP2A6*1/*4, CYP2A6*1/*9 and CYP2A6*4/*9 were 58%, 71% and 21% of the individuals genotyped as CYP2A6*1/*1, respectively. The mean in-vitro coumarin 7-hydroxylase activities in subjects carrying CYP2A6*1/*4, CYP2A6*1/*9 and CYP2A6*4/*9 were 41%, 71% and 12%, respectively, compared to those of the subjects judged as wild-type. Vmax values for coumarin 7-hydroxylation in the liver microsomes from human subjects with genotypes of CYP2A6*1/*1, CYP2A6*1/*4, CYP2A6*1/*9 and CYP2A6*4/*9 were 0.58, 0.26, 0.44 and 0.13 nmol/min/nmol total P450, respectively. CYP2A6 protein levels in human liver microsomes with the CYP2A6*4 and the CYP2A6*9 alleles were markedly decreased. These results suggest that the genetic polymorphism in the promoter region of the CYP2A6 gene (CYP2A6*9) reduced the expression levels of CYP2A6 mRNA and protein in human livers, resulting in the decrease of coumarin 7-hydroxylase activities. Individuals judged as CYP2A6*4/*9 were expected to be poor metabolizers, having extremely low activity of CYP2A6.

The synthesis and X-ray crystal structure of 9-carboxyhexahydro-7-methoxy-4a,7-ethano-benzopyran-5-en-1-one

9-Carboxyhexahydro-7-methoxy-4a,7-ethano-benzopyran-5-en-1-one (1) was prepared and examined by X-ray crystallography to probe its potential as a new peptide scaffold/template. The crystal structure of the anhydride precursor 7-(2-acetoxyethyl)-4-methoxy-3a,4,7,7a-tetrahydro-4,7-ethanoisobenzofuran-1,3-dione (6) is also reported.

Texto para discuss??o 7: Brasil s??culo XXI: a constru????o de um Estado eficaz

Em tempo algum da hist??ria a necessidade de se trabalhar a respeito da miss??o, da lideran??a e do desempenho das organiza????es sem fins lucrativos foi t??o urgente como nos dias atuais. E ?? essa urg??ncia que o presente trabalho, ???Brasil S??culo XXI ??? A constru????o de um Estado eficaz???, quer agregar ?? reflex??o da proposta de reforma do Estado brasileiro

Licitações sustentáveis: um estudo sobre a aplicação do decreto nº 7.746/2012 na contratação de serviços pela Universidade Federal do Espírito Santo – UFES

Este estudo objetiva analisar o tema Licitações Sustentáveis, as quais se destinam a integrar preocupações ambientais e sociais às aquisições públicas, no contexto do debate e da busca pela sustentabilidade. Foi realizado um estudo de caso para investigar como a Prefeitura Universitária da Universidade Federal do Espírito Santo (PU/UFES) está operacionalizando e lidando com a obrigatoriedade de adoção das licitações sustentáveis nas contratações de serviços efetuadas, considerando o período de 2010 a 2013. O principal intuito desta pesquisa foi identificar se essas contratações estão em harmonia com as diretrizes de sustentabilidade vigentes nas legislações nacionais, a partir da Instrução Normativa MPOG nº 01/2010 e, sobretudo, no que se refere ao Decreto nº 7.746/2012 e seus desdobramentos posteriores. Com isso, objetivou-se também apontar dificuldades operacionais e propor um Plano de Intervenção para melhorar a implementação de critérios sustentáveis nas referidas contratações. Em relação à metodologia utilizada, os métodos empregados foram o dedutivo e o estudo de caso. A pesquisa foi aplicada - quanto à natureza; qualitativa – quanto à forma de abordagem; descritiva e exploratória – quanto ao objetivo geral; e bibliográfica/documental – quanto aos procedimentos técnicos. O questionário/formulário e a entrevista semiestruturada foram os instrumentos utilizados para a coleta de dados. Os resultados da análise do estudo de caso realizado na PU/UFES revelam que apenas menos de um terço dos editais investigados apresentam critérios de sustentabilidade. As principais causas encontradas se dividem na falta de conhecimento dos atores envolvidos e na ausência de interesses, iniciativas e colaboração de setores relacionados. Com isso, o trabalho aponta para a necessidade de avanços em direção ao cumprimento das obrigações legais em vigor e à promoção da sustentabilidade nas contratações licitadas.

Emissões otoacústicas por produtos de distorção em crianças de 2 a 7 anos

Com a descoberta das Emissões Otoacústicas (EOA), tornou-se possível analisar e investigar as funções auditivas das células ciliadas externas do órgão da audição. Podem ser espontâneas, evocadas transientes ou por produto de distorção. O teste das EOAEs-DP caracteriza-se por ser um exame objetivo, rápido, indolor, não-invasivo e de fácil aplicação tanto para clínica como para programas de triagem auditiva. FORMA DE ESTUDO: Coorte transversal. MATERIAL E MÉTODO: Foram avaliadas 105 crianças entre 2 e 7 anos de idade da creche "Lar da Criança", de Marília, SP. A avaliação constou de exame otorrinolaringológico completo e EOAEs-DP. Todas estas crianças, após prévia autorização dos responsáveis, foram submetidas a exame otorrinolaringológico completo e EOAEs-DP. RESULTADOS: Os resultados demonstraram que das 105 crianças avaliadas, 44,76% apresentavam cerume. 12 crianças permaneceram com cerume mesmo após uso de ceruminolítico e lavagem auricular ou não apresentaram consentimento informado de seus responsáveis. Portanto, estas foram excluídas do trabalho sendo o restante, 93 crianças, submetidas a avaliação das EOAEs-DP Após a realização das EOAEs-DP, verificou-se que 5,37% das crianças apresentaram exames alterados, sendo que 60% destas eram do sexo masculino e 60% com acometimento bilateral. DISCUSSÃO: Os resultados encontrados foram inferiores aos citados na literatura, assim como o predomínio do sexo masculino. Além disso, notou-se alta prevalência de cerume nos pacientes testados. CONCLUSÃO: É essencial uma avaliação otorrinolaringológica completa prévia. O exame de EOAEs-DP pode ser realizado em crianças para detecção precoce e prevenção de falhas no desenvolvimento cognitivo e psicoemocional.

Prevalência de cárie dentária em escolares de 7 a 12 anos de idade, na cidade de Araraquara, SP (Brasil), 1979

São apresentados os resultados de levantamento epidemiológico de cárie dentária em escolares de 7 a 12 anos de idade da cidade de Araraquara, SP, Brasil, cuja água de abastecimento público é fluoretada. O objetivo do estudo é verificar a prevalência de caria dentária e o nível de assistência odontológica oferecido a esta população. A população de estudo compõe-se de escolares nascidos e sempre residentes na cidade e escolares não nascidos e/ou nem sempre residentes na cidade; são comparados os índices CPO-D obtidos para os dois grupos.

Aspectos ecológicos da Leishmaniose tegumentar americana: 7-Capacidade vetorial flebotomínea em ambiente florestal primário do Sistema da Serra do Mar, região do Vale do Ribeira, Estado de São Paulo, Brasil

Um total de 5.038 indivíduos foram coletados, na região do Vale do Ribeira, Estado de São Paulo (Brasil), durante três anos consecutivos de captura de flebotomíneo, distribuído em dezenove espécies. As armadilhas CDC instaladas na floresta contribuíram com 92,2% e a isca humana com apenas 7,0%, enquanto que no peridomicílio a CDC rendeu 0,7%. De um modo geral, as densidades obtidas com o cálculo da média geométrica de Williams foram reduzidas e a espécie mais comum na área foi P.ayrozai. O resultado de seu grau de antropofilia impede atribuir-lhe papel vetorial importante. Sendo também reduzida a presença de L.intermedia, L. migonei e L. fischeri, até na isca humana, admite-se que essas populações não estariam preenchendo condições de transmissão da doença para o homem, no ambiente florestal, ao lado de outras com hábito fundamentalmente zoófilo. O quadro mostrado poderia indicar que a função vetorial flebotomínea na região estaria sendo desenvolvida por população sobrevivente à devastação, sugerindo que um novo padrão epidemiológico da doença no Brasil parece existir e em associação com focos de L.b. braziliensis.

Studies on mosquitoes (Diptera: Culicidae) and anthropic environment: 7- Behaviour of adults Nyssorhynchus anophelines with special reference to Anopheles albitarsis s.l. in South-Eastern Brazil

Results obtained with catches performed at several sites of the Ribeira Valley, SP (Brazil) are reported. Collections with Shannon type traps showed a concentration of An.albitarsis s.l. in the Experimental Farm of Pariquera-Açu County, associated with the development of irrigated rice cultivation. Two species of the complex, named A and B, were recognized. Species B predominated in the samples. Indoor and outdoor crepuscular human bait captures were performed a the domiciliary environment. The two species showed a generally low William's media valve of 0.2, the highest value obtained was 1.6. No differences were found between indoor and outdoor behaviours between the two species of the complex.

Unesp Informa, 2010, ano 1, número 7

Boletim elaborado pela Assessoria de Comunicação e Imprensa da Reitoria da UNESP