La Imagen de China en la prensa española : una visión desde los diarios: El País, ABC, El Periódico y La Vanguardia

El presente trabajo se centra en evidenciar la valoración de China que la prensa española realiza en sus informaciones, recoger la imagen china en la misma prensa española, e identificar los mecanismos que se emplean en la construcción y difusión de esta imagen, a través de la investigación sobre la selección de las piezas informativas en los diarios de difusión nacional: El País, ABC, El Periódico y La Vanguardia, teniendo en cuenta sus distintas líneas editoriales. Acudimos principalmente a una combinación de tres metodologías: la metodología de corte documental, la cuantitativa (análisis del contenido) y la cualitativa (análisis socio-semiótico). Entre las conclusiones aportadas destacamos que, contrariamente a lo que suponíamos, en general la prensa española presenta una imagen neutral de China. Además, debido a las distintas líneas editoriales, existen varias diferencias respecto al tratamiento que realizan sobre China los perióticos objeto de estudios

IκB-ζ controls the constitutive NF-κB target gene network and survival of ABC DLBCL.

Constitutive activation of the nuclear factor-κ B (NF-κB) pathway is a hallmark of the activated B-cell-like (ABC) subtype of diffuse large B-cell lymphoma (DLBCL). Recurrent mutations of NF-κB regulators that cause constitutive activity of this oncogenic pathway have been identified. However, it remains unclear how specific target genes are regulated. We identified the atypical nuclear IκB protein IκB-ζ to be upregulated in ABC compared with germinal center B-cell-like (GCB) DLBCL primary patient samples. Knockdown of IκB-ζ by RNA interference was toxic to ABC but not to GCB DLBCL cell lines. Gene expression profiling after IκB-ζ knockdown demonstrated a significant downregulation of a large number of known NF-κB target genes, indicating an essential role of IκB-ζ in regulating a specific set of NF-κB target genes. To further investigate how IκB-ζ mediates NF-κB activity, we performed immunoprecipitations and detected a physical interaction of IκB-ζ with both p50 and p52 NF-κB subunits, indicating that IκB-ζ interacts with components of both the canonical and the noncanonical NF-κB pathway in ABC DLBCL. Collectively, our data demonstrate that IκB-ζ is essential for nuclear NF-κB activity in ABC DLBCL, and thus might represent a promising molecular target for future therapies.

Implementação do abc (activity-based costing) numa empresa prestação de serviços de saúde

O presente trabalho cujo título é Implementação do ABC numa empresa prestadora de serviços de Saúde, tem como finalidade a obtenção do grau de licenciatura em Contabilidade e Administração e tem como principal objectivo a implementação do método ABC numa pequena e média empresa de prestação de serviços de saúde, como um instrumento de apoio á gestão. Para a introdução da Contabilidade de Gestão na empresa, há que se escolher um método/sistema de apuramento de gastos que espelha a realidade da empresa, e de uma certa forma o ABC é o método ideal para apuramento de resultados sem distorções. O ABC (Activity-Based Cost) apura os resultados através da relação de causa-efeito, considerando que as actividades é que geram gastos e os objectos de custeio é que consomem as actividades. É aplicável tanto nas empresas industriais como nas empresas prestadoras de serviços, apesar de inicialmente ter sido concebido para as empresas industrias, isto é, para as grandes empresas devido aos avultados recursos financeiros e humanos como também pelo tempo necessário para a sua implementação. Mas o modelo matricial apresentado por Roztcki et al (1999) permite a aplicação deste método nas PME com poucos recursos financeiros e de tempo, utilizando uma folha de cálculo no Excel. Será este modelo a ser proposto e poderá ser implementado na clínica. O modelo apresentado foi testado num estudo de caso realizado numa clínica. Com a realização dos testes foi detectado algumas dificuldades e limitações, as maiores dificuldades encontradas foram a identificação das actividades e dos cost drivers, devido à complexidade do sector. A implementação foi concluída com sucesso, proporcionando informações detalhadas dos gastos dos produtos/serviços prestados em toda a clínica. This work was done as a requisite for obtaining a degree in Accounting and Administration, and is titled “The Implementation of ABC – Activity Based Cost in a company that provides health services”. Its main purpose is to analyze the implementation of ABC method in a small and medium-sized enterprise which provides health services to support decision making by the Managers. To adopt management accounting in a company, it’s necessary to choose a cost qualifying system that reflects the reality of the company and in a certain way ABC is the method which can determine the results without any distortion. ABC (Activity-Based Cost) determines the results through cause-and-effect relationship, whereas the activities generate spending while costing objects consume the activities. It’s applicable both in industrial companies as in services providers, although it was initially designed for industrial companies, that is, to large companies, due to the huge financial and human resources existent as well as by the time required for its implementation. But the matrix model presented by Roztckiet al (1999) allows application of this method in small and medium-sized enterprises with limited financial resources and time, using a spreadsheet in Excel. This model will be proposed and could be implemented in any clinic. The model was tested in a case study, undertaken in a private clinic. With the realization of the tests, some problems and limitations were detected, and the major difficulties encountered were the identification of activities and cost drivers, due to the complexity of the sector. The implementation was completed successfully, providing detailed information of the products services spending throughout the clinic.

Inflammatory markers and incident heart failure risk in older adults: the Health ABC (Health, Aging, and Body Composition) study.

OBJECTIVES: The purpose of this study was to evaluate the association between inflammation and heart failure (HF) risk in older adults. BACKGROUND: Inflammation is associated with HF risk factors and also directly affects myocardial function. METHODS: The association of baseline serum concentrations of interleukin (IL)-6, tumor necrosis factor-alpha, and C-reactive protein (CRP) with incident HF was assessed with Cox models among 2,610 older persons without prevalent HF enrolled in the Health ABC (Health, Aging, and Body Composition) study (age 73.6 +/- 2.9 years; 48.3% men; 59.6% white). RESULTS: During follow-up (median 9.4 years), HF developed in 311 (11.9%) participants. In models controlling for clinical characteristics, ankle-arm index, and incident coronary heart disease, doubling of IL-6, tumor necrosis factor-alpha, and CRP concentrations was associated with 29% (95% confidence interval: 13% to 47%; p < 0.001), 46% (95% confidence interval: 17% to 84%; p = 0.001), and 9% (95% confidence interval: -1% to 24%; p = 0.087) increase in HF risk, respectively. In models including all 3 markers, IL-6, and tumor necrosis factor-alpha, but not CRP, remained significant. These associations were similar across sex and race and persisted in models accounting for death as a competing event. Post-HF ejection fraction was available in 239 (76.8%) cases; inflammatory markers had stronger association with HF with preserved ejection fraction. Repeat IL-6 and CRP determinations at 1-year follow-up did not provide incremental information. Addition of IL-6 to the clinical Health ABC HF model improved model discrimination (C index from 0.717 to 0.734; p = 0.001) and fit (decreased Bayes information criterion by 17.8; p < 0.001). CONCLUSIONS: Inflammatory markers are associated with HF risk among older adults and may improve HF risk stratification.

O Sistema de Custeio “Abc” (Activity Based Costing) Como Instrumento Privilegiado de Gestão Empresarial em Cabo Verde

Com a internacionalização da economia e um mercado cada vez mais competitivo e globalizado, os gestores começam a ter a consciência de que é necessário melhorar a eficiência e reestruturar a empresa, voltando-a para eficácia, tornaram-se metas comuns no moderno ambiente de negócios. O conhecimento exacto dos custos, seu perfeito controlo e coerente mensuração passam a ser uma necessidade das empresas para que seus gerentes possam tomar decisões estratégicas e, consequentemente, fazer melhor uso dos recursos organizacionais, cada vez mais limitados. Para atender a esses anseios, as organizações vêm buscando, cada vez mais, utilizar o sistema de custeio ABC (Activity Based Costing) ou custeio baseado em actividades, como sendo, um método de apuramento de custos que tem vindo a ganhar alguma popularidade nos últimos tempos. Este método fornece informação mais correcta e precisa sobre os custos, a qual é extremamente útil para auxiliar os gestores na tomada de decisões. Apesar de ser um conceito universal, a implementação bem sucedida do ABC não é igual em todas as organizações e deve ser adaptada a uma estratégia, estrutura, capacidade e necessidade únicas da empresa. O object ivo deste trabalho é aprofundar o conhecimento sobre este método “ABC”, estudando a sua aplicabilidade nas empresas de Cabo Verde, mas concretamente na cidade da Praia. Pretende-se, também, identificar as dificuldades no decorrer da sua implementação e averiguar se as empresas que adoptam ou estão a adoptar o ABC consideram-no uma ferramenta de gestão empresarial.

O Sistema Integrado ABC-EVA como ferramenta para a Tomada de Decisão

O sistema Activity Based Costing (ABC) surgiu em meados da década de 80 pela iniciativa dos autores Robin Kaplan, Robin Cooper e Thomas Johnson, com o intuito de suprir as deficiências apresentadas pelos sistemas de custeio tradicionais na imputação arbitrária dos custos indirectos. Alvo de vários elogios e críticas, o sistema ABC é, actualmente, percebido por muitos como a forma mais apropriada de calcular o custo dos produtos. No entanto, numa altura em que é dada cada vez mais importância à criação de valor, especialmente à criação de valor aos accionistas, o sistema ABC se demonstrou insuficiente, pois não considera o Custo de Oportunidade do Capital (COC) investido pelos accionistas. Neste sentido, autores como Narcyz Roztocki e Kim LaScola Needy vieram chamar a atenção desse facto e proporam a integração do sistema ABC ao indicador do desempenho Economic Value Added (EVA) como forma de responder a essa limitação visto que este indicador considera o custo do capital investido pelos accionistas. O estudo de caso ora apresentado visou compreender os fundamentos do sistema integrado ABC-EVA e, compreender os impactos desse sistema no processo decisório, através da sua implementação numa empresa do país. Para a implementação do sistema seguimos os passos enumerados por Roztocki et al. (2004) e Roztocki e Needy (1999c), para a implementação do sistema ABC e para a implementação do sistema integrado ABC-EVA, respectivamente. Dos resultados do estudo pudemos concluir que o sistema integrado ABC-EVA dota os gestores de uma informação mais completa sobre os custos e, por isso, é mais apropriada à tomada de decisões a nível de preços, custos, política de marketing, entre outros. Não obstante, o estudo de caso apresentou algumas limitações que, ainda que tenham condicionado as conclusões do estudo, acabaram por comprovar os resultados de alguns estudos referidos na revisão da literatura. Espera-se que a empresa objecto de estudo possa beneficiar, de alguma forma, do estudo realizado e que, se interesse em adoptar o sistema proposto ou outro, já que não possui nenhum.

Regional variations in ABC transporter expression along the mouse intestinal tract.

The ATP-binding cassette (ABC) family of proteins comprise a group of membrane transporters involved in the transport of a wide variety of compounds, such as xenobiotics, vitamins, lipids, amino acids, and carbohydrates. Determining their regional expression patterns along the intestinal tract will further characterize their transport functions in the gut. The mRNA expression levels of murine ABC transporters in the duodenum, jejunum, ileum, and colon were examined using the Affymetrix MuU74v2 GeneChip set. Eight ABC transporters (Abcb2, Abcb3, Abcb9, Abcc3, Abcc6, Abcd1, Abcg5, and Abcg8) displayed significant differential gene expression along the intestinal tract, as determined by two statistical models (a global error assessment model and a classic ANOVA, both with a P < 0.01). Concordance with semiquantitative real-time PCR was high. Analyzing the promoters of the differentially expressed ABC transporters did not identify common transcriptional motifs between family members or with other genes; however, the expression profile for Abcb9 was highly correlated with fibulin-1, and both genes share a common complex promoter model involving the NFkappaB, zinc binding protein factor (ZBPF), GC-box factors SP1/GC (SP1F), and early growth response factor (EGRF) transcription binding motifs. The cellular location of another of the differentially expressed ABC transporters, Abcc3, was examined by immunohistochemistry. Staining revealed that the protein is consistently expressed in the basolateral compartment of enterocytes along the anterior-posterior axis of the intestine. Furthermore, the intensity of the staining pattern is concordant with the expression profile. This agrees with previous findings in which the mRNA, protein, and transport function of Abcc3 were increased in the rat distal intestine. These data reveal regional differences in gene expression profiles along the intestinal tract and demonstrate that a complete understanding of intestinal ABC transporter function can only be achieved by examining the physiologically distinct regions of the gut.

IMPORTÂNCIA E UTILIZAÇÃO DO MÉTODO ABC (ACTIVITY BASED COSTING) COMO SISTEMA DE CUSTEIO – ANALISE DA SITUAÇAO NAS EMPRESAS INDUSTRIAIS DA CIDADE DA PRAIA

O presente trabalho cujo tema é Importância e Utilização de Método ABC (Actvity Based Costing) como Sistema de Custeio, tem como objetivo principal analisar a importância do método ABC no processo de tomada de decisão por parte dos gestores das empresas. Kaplan e Cooper, na década de 80 do século passado, demonstraram que os sistemas de custeio tradicionais são insuficientes para satisfazer as necessidades atuais das empresas, visto que um dos maiores problemas atualmente enfrentados pelas empresas em relação ao custeio dos produtos é a dimensão sempre crescente dos custos indiretos. Por outro lado, os sistemas tradicionais de apuramento de custo não imputam adequadamente os custos indiretos da produção aos produtos, dificultando uma análise de custos. E diante disso, criaram o sistema de custeio baseado na atividade. O ABC parte de um princípio de base muito simples: os recursos são consumidos por atividades e estas são consumidas pelos objetos de custeio. Este sistema de custeio veio dinamizar e dar outra credibilidade à Contabilidade de Gestão, pela importância que dá às atividades e pela visão da organização e do método de gestão, permite em princípio, um mais exato custeio dos produtos e uma mais apurada avaliação de desempenho. Ao identificar atividades que não acrescentam valor, vem facultar informação que permite uma reorganização da estrutura da empresa, e a sua consequente melhoria em termos de rentabilidade e competitividade. O presente estudo resulta do levantamento realizado e das análises das informações e dados das grandes e médias empresas industriais da Cidade da Praia, através dum inquérito por questionário. De acordo com o resultado obtido, concluiu-se que a maioria das grandes e médias empresas industriais da Cidade da Praia não utilizam o método ABC devido ao desconhecimento desta técnica e também verificou-se que o ABC é um importante instrumento de tomada de decisão por parte dos gestores das empresas.

La tendencia del periodismo tradicional a la cultura light en las portadas de El País y ABC

El present projecte analitza el panorama de la premsa tradicional espanyola a partir dels anys 60 fins als últims dies. El treball proposa un estudi sobre la tendència dels periòdics cap a una cultura light caracteritzada per la seva levitat i la creixent aposta per l'espectacle. Així mateix, es reflexiona sobre el grau d'importància de les relacions públiques com a fonts informatives per als mitjans de comunicació. Aquest escenari mediàtic es posa en pràctica contrastant una varietat de portades de l'ABC i El País a partir del seu primer punt de trobada.

Método de determinação de enxofre lábil com membrana de troca aniônica

A disponibilidade de enxofre no solo pode ser explicada pela inter-relação dos fatores intensidade, quantidade e capacidade-tampão. O fator quantidade refere-se à reserva lábil do nutriente, ou seja, a quantidade que, presente na fase sólida do solo, possa passar à solução, no tempo necessário para o ciclo vital de uma cultura de ciclo curto ou anual. Com o objetivo de desenvolver um método para determinar a quantidade de enxofre lábil do solo, utilizaram-se amostras de três solos que foram incubadas, por 60 dias, após receberem os seguintes tratamentos: (a) amostra não desinfetada e sem adição de enxofre (-D, -S); (b) amostra desinfetada sem adição de enxofre (+D, -S); (c) amostra não desinfetada com adição de enxofre (-D, +S), e (d) amostra desinfetada e com adição de enxofre (+D, +S). Para a determinação do enxofre lábil, foram utilizadas membranas de troca aniônica de 50 x 20 mm saturadas com o íon Cl-. Para definir o método de determinação do enxofre lábil, realizaram-se três ensaios, sendo: (1) extração com diferente número de membranas de troca aniônica; (2) extrações sucessivas de 16 h cada, com uma membrana, e (3) extração com uma membrana por diferentes tempos de agitação (1, 2, 4, 8, 16, 24, 36, 48 e 72 h). Os resultados indicaram que a extração com agitação contínua por 48 h com uma membrana de troca aniônica constitui método viável e eficiente, tendo extraído uma elevada percentagem do enxofre lábil, denominado enxofre rapidamente lábil (SRL). O enxofre extraído entre 48 e 152 h foi chamado de enxofre lentamente lábil (SLL), enquanto o enxofre do solo que não foi extraído com um tempo de agitação de até 152 h foi denominado não-lábil (SNL).

Integrated Analysis of High-Resolution Gene Expression and Copy Number Profiling Identified Biallelic Deletion of CDKN2A/2B Tumor Suppressor Locus As the Most Frequent and Unique Genomic Abnormality in Diffuse Large B-Cell Lymphoma (DLBCL) with Strong Prognostic Value in Both GCB and ABC Subtypes and Not Overcome by a Dose-Intensive Immunochemotherapy Regimen Plus Rituximab. Results of a Prospective GELA Clinical Trial Program

Background and aim of the study: Genomic gains and losses play a crucial role in the development and progression of DLBCL and are closely related to gene expression profiles (GEP), including the germinal center B-cell like (GCB) and activated B-cell like (ABC) cell of origin (COO) molecular signatures. To identify new oncogenes or tumor suppressor genes (TSG) involved in DLBCL pathogenesis and to determine their prognostic values, an integrated analysis of high-resolution gene expression and copy number profiling was performed. Patients and methods: Two hundred and eight adult patients with de novo CD20+ DLBCL enrolled in the prospective multicentric randomized LNH-03 GELA trials (LNH03-1B, -2B, -3B, 39B, -5B, -6B, -7B) with available frozen tumour samples, centralized reviewing and adequate DNA/RNA quality were selected. 116 patients were treated by Rituximab(R)-CHOP/R-miniCHOP and 92 patients were treated by the high dose (R)-ACVBP regimen dedicated to patients younger than 60 years (y) in frontline. Tumour samples were simultaneously analysed by high resolution comparative genomic hybridization (CGH, Agilent, 144K) and gene expression arrays (Affymetrix, U133+2). Minimal common regions (MCR), as defined by segments that affect the same chromosomal region in different cases, were delineated. Gene expression and MCR data sets were merged using Gene expression and dosage integrator algorithm (GEDI, Lenz et al. PNAS 2008) to identify new potential driver genes. Results: A total of 1363 recurrent (defined by a penetrance > 5%) MCRs within the DLBCL data set, ranging in size from 386 bp, affecting a single gene, to more than 24 Mb were identified by CGH. Of these MCRs, 756 (55%) showed a significant association with gene expression: 396 (59%) gains, 354 (52%) single-copy deletions, and 6 (67%) homozygous deletions. By this integrated approach, in addition to previously reported genes (CDKN2A/2B, PTEN, DLEU2, TNFAIP3, B2M, CD58, TNFRSF14, FOXP1, REL...), several genes targeted by gene copy abnormalities with a dosage effect and potential physiopathological impact were identified, including genes with TSG activity involved in cell cycle (HACE1, CDKN2C) immune response (CD68, CD177, CD70, TNFSF9, IRAK2), DNA integrity (XRCC2, BRCA1, NCOR1, NF1, FHIT) or oncogenic functions (CD79b, PTPRT, MALT1, AUTS2, MCL1, PTTG1...) with distinct distribution according to COO signature. The CDKN2A/2B tumor suppressor locus (9p21) was deleted homozygously in 27% of cases and hemizygously in 9% of cases. Biallelic loss was observed in 49% of ABC DLBCL and in 10% of GCB DLBCL. This deletion was strongly correlated to age and associated to a limited number of additional genetic abnormalities including trisomy 3, 18 and short gains/losses of Chr. 1, 2, 19 regions (FDR < 0.01), allowing to identify genes that may have synergistic effects with CDKN2A/2B inactivation. With a median follow-up of 42.9 months, only CDKN2A/2B biallelic deletion strongly correlates (FDR p.value < 0.01) to a poor outcome in the entire cohort (4y PFS = 44% [32-61] respectively vs. 74% [66-82] for patients in germline configuration; 4y OS = 53% [39-72] vs 83% [76-90]). In a Cox proportional hazard prediction of the PFS, CDKN2A/2B deletion remains predictive (HR = 1.9 [1.1-3.2], p = 0.02) when combined with IPI (HR = 2.4 [1.4-4.1], p = 0.001) and GCB status (HR = 1.3 [0.8-2.3], p = 0.31). This difference remains predictive in the subgroup of patients treated by R-CHOP (4y PFS = 43% [29-63] vs. 66% [55-78], p=0.02), in patients treated by R-ACVBP (4y PFS = 49% [28-84] vs. 83% [74-92], p=0.003), and in GCB (4y PFS = 50% [27-93] vs. 81% [73-90], p=0.02), or ABC/unclassified (5y PFS = 42% [28-61] vs. 67% [55-82] p = 0.009) molecular subtypes (Figure 1). Conclusion: We report for the first time an integrated genetic analysis of a large cohort of DLBCL patients included in a prospective multicentric clinical trial program allowing identifying new potential driver genes with pathogenic impact. However CDKN2A/2B deletion constitutes the strongest and unique prognostic factor of chemoresistance to R-CHOP, regardless the COO signature, which is not overcome by a more intensified immunochemotherapy. Patients displaying this frequent genomic abnormality warrant new and dedicated therapeutic approaches.

Rapid radiation in spiny lobsters (Palinurus spp) as revealed by classic and ABC methods using mtDNA and microsatellite data

Background: Molecular tools may help to uncover closely related and still diverging species from a wide variety of taxa and provide insight into the mechanisms, pace and geography of marine speciation. There is a certain controversy on the phylogeography and speciation modes of species-groups with an Eastern Atlantic-Western Indian Ocean distribution, with previous studies suggesting that older events (Miocene) and/or more recent (Pleistocene) oceanographic processes could have influenced the phylogeny of marine taxa. The spiny lobster genus Palinurus allows for testing among speciation hypotheses, since it has a particular distribution with two groups of three species each in the Northeastern Atlantic (P. elephas, P. mauritanicus and P. charlestoni) and Southeastern Atlantic and Southwestern Indian Oceans (P. gilchristi, P. delagoae and P. barbarae). In the present study, we obtain a more complete understanding of the phylogenetic relationships among these species through a combined dataset with both nuclear and mitochondrial markers, by testing alternative hypotheses on both the mutation rate and tree topology under the recently developed approximate Bayesian computation (ABC) methods. Results Our analyses support a North-to-South speciation pattern in Palinurus with all the South-African species forming a monophyletic clade nested within the Northern Hemisphere species. Coalescent-based ABC methods allowed us to reject the previously proposed hypothesis of a Middle Miocene speciation event related with the closure of the Tethyan Seaway. Instead, divergence times obtained for Palinurus species using the combined mtDNA-microsatellite dataset and standard mutation rates for mtDNA agree with known glaciation-related processes occurring during the last 2 my. Conclusion The Palinurus speciation pattern is a typical example of a series of rapid speciation events occurring within a group, with very short branches separating different species. Our results support the hypothesis that recent climate change-related oceanographic processes have influenced the phylogeny of marine taxa, with most Palinurus species originating during the last two million years. The present study highlights the value of new coalescent-based statistical methods such as ABC for testing different speciation hypotheses using molecular data.

The half-size ABC transporters STR1 and STR2 are indispensable for mycorrhizal arbuscule formation in rice.

The central structure of the symbiotic association between plants and arbuscular mycorrhizal (AM) fungi is the fungal arbuscule that delivers minerals to the plant. Our earlier transcriptome analyses identified two half-size ABCG transporters that displayed enhanced mRNA levels in mycorrhizal roots. We now show specific transcript accumulation in arbusculated cells of both genes during symbiosis. Presently, arbuscule-relevant factors from monocotyledons have not been reported. Mutation of either of the Oryza sativa (rice) ABCG transporters blocked arbuscule growth of different AM fungi at a small and stunted stage, recapitulating the phenotype of Medicago truncatula stunted arbuscule 1 and 2 (str1 and str2) mutants that are deficient in homologous ABCG genes. This phenotypic resemblance and phylogenetic analysis suggest functional conservation of STR1 and STR2 across the angiosperms. Malnutrition of the fungus underlying limited arbuscular growth was excluded by the absence of complementation of the str1 phenotype by wild-type nurse plants. Furthermore, plant AM signaling was found to be intact, as arbuscule-induced marker transcript accumulation was not affected in str1 mutants. Strigolactones have previously been hypothesized to operate as intracellular hyphal branching signals and possible substrates of STR1 and STR2. However, full arbuscule development in the strigolactone biosynthesis mutants d10 and d17 suggested strigolactones to be unlikely substrates of STR1/STR2. Interestingly, rice STR1 is associated with a cis-natural antisense transcript (antiSTR1). Analogous to STR1 and STR2, at the root cortex level, the antiSTR1 transcript is specifically detected in arbusculated cells, suggesting unexpected modes of STR1 regulation in rice.

The role of the vacuolar H+ - ATPase in membrane fusion

RésuméLa H+-ATPase vacuolaire (V-ATPase) est un complexe enzymatique composé de deux secteurs multimériques (VQ et Vi) dont l'association dans la cellule est réversible. Le secteur intramembranaire de la V-ATPase (V0) interagit physiquement avec des protéines SNARE et stimule la fusion homotypique des vacuoles de la levure (lysosomes), la sécrétion de neurotransmetteurs et d'insuline, la fusion entre phagosome et lysosome ainsi que la sécrétion des corps multivésiculaires par un mécanisme inconnu. Dans cette étude j'ai identifié des résidues d'acides amines situés dans des sous-unités de V0 impliqués dans le mécanisme de fusion des vacuoles mais non essentiels pour l'acidification vacuolaire par la V-ATPase. j'ai utilisé un protocole de mutagenèse aléatoire pour produire des libraries de mutants des sous unités de V0. Ces libraries ont été analysées in vivo afin d'identifier des alleles qui permettent la translocation des protons mais produisent une vacuole fragmentée, phénotype indiquant un défaut dans la fusion membranaire. Les vacuoles des mutants ont été isolées et caractéisées en utilisant une grande variété d'outils biochimiques pour déterminer précisément l'impact des différentes mutations sur l'accomplissement d'événements clés du processus de fusion.J'ai identifié des mutations associées à des défauts spécifiques de la fusion dans plusieurs sous-unités de V0. Dans les protéolipides c, c' et c" ces mutations se concentrent dans la partie cytosolique des domaines transmembranaires. Elles renforcent les associations entre les secteurs de la V-ATPase et entre V0 et les SNAREs. Dans la fusion vacuolaire ces mutations permettent la formation de complexes SNAREs en trans mais inhibent l'induction de la fusion. Par contre, la deletion de la sous- unité d influence les étapes de la fusion qui précèdent la formation des complexes trans-SNAREs. Mes résultats démontrent que V0 joue des rôles différents dans plusieurs étapes de la fusion et que ces fonctions sont liées au système des SNAREs. Ils différencient génétiquement les activités de V0 dans la translocation des protons et dans la fusion et identifient de nombreux résidus importants pour la fusion vacuolaire. De plus, compte tenu de la grande conservation de sequence des protéolipides chez les eukaryotes les mutations identifiées dans cette l'étude apportent de nouvelles informations pour analyser la fonction de V0 dans des organismes multicellulaires pour lesquels la function catalytique de la V-ATPase est essentielle à la survie.Résumé pour le large publicLe transport de protéines et de membranes est important pour maintenir la fonction des organelles dans la cellule. Il s'excerce au niveau des vesicules. La fusion membranaire est un processus élémentaire de ce transport. Pour fusionner deux membranes, il faut la coordination de deux activités: le rapprochement et la déstabiiization des deux membranes. La collaboration d'un ensemble de proteins conservés chez les eukaryotes, est nécessaire pour catalyser ces activités. Les proteins SNAREs sont les protagonistes principaux dans la fusion membranaire. Néanmoins, d'autres protéines, comme des Rab-GTPases et des chaperonnes, sont nécessaires pour permettre ce phénomène de fusion. Toutes ces protéines sont temporairement associées avec les SNAREs et leur fonction dans la fusion membranaire est souvent directement liée à leur activité dans cette association. Le secteur transmembranaire V0 de la V-ATPase rnteragit avec des SNAREs et est essentiel pour la fusion dans une variété de systèmes modèles comme la mouche, la souris et la levure. Le secteur V0 est composé de six protéines différentes. Avec te secteur Va, qui réside dans le cytosol, il forme la V-ATPase dont la fonction principale est l'acidification des organelles par translocation des protons à travers la membrane par un mécanisme ressemblant à celui d'une pompe. V0joue un role dans la fusion membranaire, indépendamment de son activité catalytique liée au pompage des protons, et ce rôle est encore largement méconnu à ce jour. Le but de ma thèse était de mieux comprendre l'implication de V0 dans ce contexte.Pour étudier des activités liées à la V-ATPase, la levure est un excellent modèle d'étude car elle survie à une inactivation de l'enzyme alors que le meme traitement serait léthal pour des organismes multicellulaires. Dans ma thèse j'ai utilisé la fusion homotypique de la vacuole de levure comme système modèle pour étudier le rôle de V0 dans la fusion. J'ai muté des gènes qui encodent des sous- unités de V0 et les ai introduit dans des souches privées des gènes respectifs. Dans les librairies de souches portant différentes versions de ces gènes j'ai cherché des clones exprimant une V-ATPase intacte et fonctionnelle mais qui possèdent une vacuole fragmentée. Le plus souvent, une vacuole fragmentée indique un défaut dans la fusion vacuolaire. Dans les trois types de protéolipides qui composent un cylindre dans le secteur V0, j'ai trouvé des clones avec une vacuole fragmentée. Après avoir isolé les mutations responsable de ce type de morphologie vacuolaire, j'ai isolé les vacuoles de ces clones pour étudier leur activités dans différentes étapes de la fusion vacuolaire. Les résultats de ces analyses mettent en évidence une implication de V0 dans plusieurs étapes de la fusion vacuolaire. Certaines mutations sélectionnées dans mon étude inhibent une étape précoce de la fusion qui inclue la dissociation des complexes SNARE, tandis que d'autres mutations inhibent une étape tardive du processus de fusion qui inclue la transmission d'une force disruptive dans la membrane.AbstractThe membrane-integral V0 sector of the vacuolar H+-ATPase (V-ATPase) interacts with SNARE proteins. V0 stimulates fusion between yeast vacuoles (lysosomes) (Peters et al., 2001b), secretion of neurotransmitters and insulin (Hiesinger et al., 2005a, Sun-Wada et al., 2006a), phagosome-lysosome fusion (Peri and Nusslein-Volhard, 2008) and secretion of multivesicular bodies (Liegeois et al., 2006b) by a yet unknown mechanism. In my thesis, I identified sites in V0 subunits that are involved in yeast vacuole fusion but dispensable for the proton pumping by the V-ATPase. I randomly mutagenized V0 subunits and screened in vivo for mutant alleles that support proton pumping but cause fragmented vacuoles, a phenotype indicative of a fusion defect. Mutant vacuoles were isolated and analyzed in a cell-free system, allowing assay of key events in fusion, such as trans-SNARE pairing, lipid transition and fusion pore opening (Reese et al., 2005b).Mutants with selective fusion defects were found in several V0 subunits. In the proteolipids c, c' and c", critical mutations are concentated in the cytosolic half of the transmembrane domains. These mutations rendered the V-ATPase holoenzyme more stable and modulated V0-SNARE associations. In vacuole fusion critical proteolipid mutations permitted trans-SNARE pairing but impeded the induction of lipid flow between the membranes. Deletion of subunit d, by contrast, influenced early stages of fusion that precede trans-SNARE pairing. My results show that V0 acts in several steps of the fusion process and that its function is intimately connected to the SNARE system. They genetically separate the proton pump and fusion activities of V0 and identify numerous critical residues. Given the high sequence conservation of proteolipids in eukaryotic life, the identified mutations may be helpful in analyzing the fusion function of V0 also in mammalian cells, where V- ATPase pump function is essential for survival.