Tempo de início de tratamento de pessoas vivendo com HIV, por nivel de complexidade, no município do Rio de Janeiro, 2014-2016

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Ciências da Saúde, Programa de Pós-Graduação em Saúde Coletiva, 2018.Introdução: o cuidado integral e o início do tratamento oportuno são fatores essenciais para o sucesso terapêutico das pessoas vivendo com HIV (PVHIV). Diferentes níveis de complexidade de serviços de saúde podem oferecer tratamento segundo as especificidades do cuidado dispensado, a saber: a atenção primária em saúde (APS), a secundária e a hospitalar. Objetivos: comparar o tempo de início do tratamento antirretroviral, medido pelo intervalo em dias entre o primeiro exame de carga viral do HIV (CV-HIV) e/ou da contagem de linfócitos T-CD4+ (LT-CD4+), e o início da terapia antirretroviral (TARV), em diferentes níveis de complexidade da atenção em saúde, no município do Rio de Janeiro. Método: estudo de coorte histórica, que utiliza a análise de sobrevida. Após estimativa da hazard ratio mediante a regressão de Cox, foi utilizado o teste AIC para determinar o melhor modelo. A principal variável explicativa foi o nível de complexidade dos serviços; também foram avaliados fatores sociodemográficos e clínicos disponíveis nas bases de dados relacionadas. Resultados: foram incluídas no estudo 6.963 PVHIV com 18 anos e mais. Cerca de 69% eram do sexo masculino; ~49% tinham entre 25 e 39 anos; ~61% apresentaram CV≥10.000 cópias/mL; ~39% tiveram contagem de CD4≥500 células/mm3 e ~46% eram atendidas na APS. A variável que mostrou maior força de associação com o tempo para início de tratamento foi a CV-HIV (CV≥10.000; HR=6,3; IC 95%=5,3-7,5). A análise demonstrou que o início do tratamento ocorria em menor tempo para as pessoas com piores condições clínicas. O tempo para início de tratamento foi decrescente, mas ainda se apresentou tardio, tanto para homens quanto para mulheres (84 e 108 dias, respectivamente). Nas variáveis sociodemográficas e clínicas, observou-se que jovens e pessoas com maior escolaridade eram atendidas em menor tempo. As pessoas com maior idade, com menor escolaridade e autodeclaradas de cor preta registraram as maiores medianas de tempo para início da TARV. Conclusões: este estudo sustenta que o tempo para início de tratamento das PVHIV na APS é significativamente menor do que na atenção hospitalar. Contudo, não difere do tempo para início de tratamento na atenção especializada. A oferta de tratamento de PVHIV na atenção primária não apresenta prejuízo em relação ao tempo para início do tratamento, se comparada aos outros níveis de complexidade.Introduction: Integral care and timely initiation of antiretroviral treatment (ART) are crucial factors for therapeutic success of people living with HIV (PLHIV). Health services with different levels of complexity can offer treatment according to the specificities of the care provided, ranging from primary health care (PHC), to secondary and hospital-based care. Objectives: To compare time of ART initiation, as measured by the interval in days between the first laboratory diagnosis (HIV viral load test - CV-HIV and / or CD4+ T-cell count), and the beginning of ART, among different levels of complexity of health care, in the city of Rio de Janeiro, Brazil. Method: Historical cohort study using secondary data for survival analysis. After estimating the hazard ratio using Cox regression, the AIC test was used to determine the best model fit. The outcome variable was the time for ART initiation; the main explanatory variable was the level of complexity of the services. Sociodemographic and clinical factors were also evaluated in the related databases. Results: 6,963 PLWHIV aged 18 years and over were included in the study. About 69% were male; 48% were aged 25 to 39 years; 60% had CV≥10,000 copies / ml; 39% had CD4≥500 cells / mm3 and ~46% were treated at the PHC. The variable that showed the greatest strength of association with the time for treatment initiation was CV-HIV (CV≥10.000, HR=6.3, 95% CI=5.3-7.5). Analysis showed that the time for treatment beginning was shorter for people with worse clinical conditions. Median time to start treatment decreased, but was still delayed for both men and women (84 and 108 days, respectively). Regarding sociodemographic and clinical variables, we observed that young people and people with higher schooling had their treatment initiated in a shorter time. Older people, with lower schooling and those self-declared black registered the highest medians of time to start ART. Conclusions: This study sustains that the time for ART initiation to PLHIV in PHC is significantly lower than in hospital care. However, it does not differ from the time to start treatment in specialized care; thus, PHC is not worse than other levels of complexity

Fairness protocols for optical ring networks

Orientadores: Nelson Luis Saldanha da Fonseca, Marcos Rogerio SalvadorDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de ComputaçãoResumo: Em redes ópticas em anel com slotting, slots de tamanhos fixos circulam continuamente pelo anel transportando pacotes de dados entre os nós. Em tais redes, um slot, ocupado com pacotes de um nó, tem seu conteúdo removido somente pelo próprio nó de origem. Entretanto, é possível remover o conteúdo do slot no nó de destino, técnica conhecida como remoção no destino, o que permite que um slot seja utilizado mais de uma vez em cada ciclo, o que é conhecido como reutilização espacial. Esta prática aumenta consideravelmente a vazão do anel. Entretanto, se o acesso aos slots não for controlado, injustiças podem ocorrer devido a oportunidades desbalanceadas de acesso ao meio oferecido aos nós. Para previnir um acesso injusto ao meio, a ocupação dos slots é controlada pelos protocolos de controle de acesso ao meio (MAC), que distribuem os slots entre os nós, oferecendo oportunidades justas do acesso ao meio. Os protocolos MAC seguem políticas de justiça, que são regras que determinam a divisão justa dos recursos do anel. Os protocolos MAC que oferecem justiça entre nós são comuns na literatura, entretanto, estes protocolos ignoram a justiça entre conexões TCP (Transmission Control Protocol)o Nesta dissertação, três novos protocolos são apresentados: LCR-SD, TCP-Fair e RVQ. O protocolo LCR-SD distribui a largura de banda baseada na política de justiça entre pares origem-destino, o protocolo RVQ oferece justiça entre conexões TCP e o protocolo de TCP -Fair oferece a justiça entre as conexões TCP mantendo a justiça entre nós. Os protocolos são comparados através de simulações realizadas no Network Simulator (NS-2). Resultados indicam que os protocolos LCR-SD, TCP-Fair e RVQ oferecem uma vazão superior ao protocolo Metaring. Além disso, apresenta-se um estudo sobre o impacto do tamanho dos slots no transporte de tráfego da InternetAbstract: In slotted ring networks, slots of fixed size continuously circulate the ring transferring data packets between nodes. In such networks, a slot occupied by packets from one node, has its content removed only by the source node. However, it is possible to remove the packet content in the destination node, technique known as destination removal, which allows a slot to be used more than once in each cycle, leading to spatial reuse, which increases considerably the throughput. However, if the access to the slots is not regulated, unfairness may occur due to unbalanced medium access opportunities offered to the nodes. To prevent unfair access to the medium, the occupation of the slots is regulated by a Medium Access Control protocol (MAC), which distributes the slots among the nodes, offering fair access opportunities to the medium. MAC protocols comply with fairness policies, which are rules that determine the fair distribution of the ring resources. MAC protocols that offer fairness among nodes are common in the literature, however, these protocols ignore the fairness among TCP (Transmission Control Protocol) connections. In this dissertation, three new protocols are presented: LCR-SD, TCP-Fair and RVQ. The LCR-SD protocol distributes the bandwidth based on the source-destination node fairness policy, the RVQ protocol offers fairness among TCP connections and the TCP-Fair protocol offers fairness among TCP connections and maintains fairness among nodes. The protocols are compared through simulations using the Network Simulator (NS- 2). Results indicate that the protocols LCR-SD, TCP-Fair and RVQ offer a superior throughput compared to the Metaring protocol. Moreover, a study on the impact of the slot size on the transport of Internet traffic is presented.MestradoRedes de ComputadoresMestre em Ciência da Computaçã

The yeast Cbk1 kinase regulates mRNA localization via the mRNA-binding protein Ssd1

In the absence of Cbk1 phosphorylation Ssd1-associated mRNAs are redirected from sites of polarized cell growth to stress granules and P-bodies

Novel activity of eukaryotic translocase, eEF2: dissociation of the 80S ribosome into subunits with ATP but not with GTP

Ribosomes must dissociate into subunits in order to begin protein biosynthesis. The enzymes that catalyze this fundamental process in eukaryotes remained unknown. Here, we demonstrate that eukaryotic translocase, eEF2, which catalyzes peptide elongation in the presence of GTP, dissociates yeast 80S ribosomes into subunits in the presence of ATP but not GTP or other nucleoside triphosphates. Dissociation was detected by light scattering or ultracentrifugation after the split subunits were stabilized. ATP was hydrolyzed during the eEF2-dependent dissociation, while a non-hydrolyzable analog of ATP was inactive in ribosome splitting by eEF2. GTP inhibited not only ATP hydrolysis but also dissociation. Sordarin, a fungal eEF2 inhibitor, averted the splitting but stimulated ATP hydrolysis. Another elongation inhibitor, cycloheximide, also prevented eEF2/ATP-dependent splitting, while the inhibitory effect of fusidic acid on the splitting was nominal. Upon dissociation of the 80S ribosome, eEF2 was found on the subunits. We propose that the dissociation activity of eEF2/ATP plays a role in mobilizing 80S ribosomes for protein synthesis during the shift up of physiological conditions

The distribution of active RNA polymerase II along the transcribed region is gene-specific and controlled by elongation factors

In order to study the intragenic profiles of active transcription, we determined the relative levels of active RNA polymerase II present at the 3′- and 5′-ends of 261 yeast genes by run-on. The results obtained indicate that the 3′/5′ run-on ratio varies among the genes studied by over 12 log2 units. This ratio seems to be an intrinsic characteristic of each transcriptional unit and does not significantly correlate with gene length, G + C content or level of expression. The correlation between the 3′/5′ RNA polymerase II ratios measured by run-on and those obtained by chromatin immunoprecipitation is poor, although the genes encoding ribosomal proteins present exceptionally low ratios in both cases. We detected a subset of elongation-related factors that are important for maintaining the wild-type profiles of active transcription, including DSIF, Mediator, factors related to the methylation of histone H3-lysine 4, the Bur CDK and the RNA polymerase II subunit Rpb9. We conducted a more detailed investigation of the alterations caused by rpb9Δ to find that Rpb9 contributes to the intragenic profiles of active transcription by influencing the probability of arrest of RNA polymerase II

Membrane-association of mRNA decapping factors is independent of stress in budding yeast

Recent evidence has suggested that the degradation of mRNA occurs on translating ribosomes or alternatively within RNA granules called P bodies, which are aggregates whose core constituents are mRNA decay proteins and RNA. In this study, we examined the mRNA decapping proteins, Dcp1, Dcp2, and Dhh1, using subcellular fractionation. We found that decapping factors co-sediment in the polysome fraction of a sucrose gradient and do not alter their behaviour with stress, inhibition of translation or inhibition of the P body formation. Importantly, their localisation to the polysome fraction is independent of the RNA, suggesting that these factors may be constitutively localised to the polysome. Conversely, polysomal and post-polysomal sedimentation of the decapping proteins was abolished with the addition of a detergent, which shifts the factors to the non-translating RNP fraction and is consistent with membrane association. Using a membrane flotation assay, we observed the mRNA decapping factors in the lower density fractions at the buoyant density of membrane-associated proteins. These observations provide further evidence that mRNA decapping factors interact with subcellular membranes, and we suggest a model in which the mRNA decapping factors interact with membranes to facilitate regulation of mRNA degradation

An mRNA decapping mutant deficient in P body assembly limits mRNA stabilization in response to osmotic stress

Yeast is exposed to changing environmental conditions and must adapt its genetic program to provide a homeostatic intracellular environment. An important stress for yeast in the wild is high osmolarity. A key response to this stress is increased mRNA stability primarily by the inhibition of deadenylation. We previously demonstrated that mutations in decapping activators (edc3∆ lsm4∆C), which result in defects in P body assembly, can destabilize mRNA under unstressed conditions. We wished to examine whether mRNA would be destabilized in the edc3∆ lsm4∆C mutant as compared to the wild-type in response to osmotic stress, when P bodies are intense and numerous. Our results show that the edc3∆ lsm4∆C mutant limits the mRNA stability in response to osmotic stress, while the magnitude of stabilization was similar as compared to the wild-type. The reduced mRNA stability in the edc3∆ lsm4∆C mutant was correlated with a shorter PGK1 poly(A) tail. Similarly, the MFA2 mRNA was more rapidly deadenylated as well as significantly stabilized in the ccr4∆ deadenylation mutant in the edc3∆ lsm4∆C background. These results suggest a role for these decapping factors in stabilizing mRNA and may implicate P bodies as sites of reduced mRNA degradation

Nonsense-mediated mRNA decay controls the changes in yeast ribosomal protein pre-mRNAs levels upon osmotic stress

The expression of ribosomal protein (RP) genes requires a substantial part of cellular transcription, processing and translation resources. Thus, the RP expression must be tightly regulated in response to conditions that compromise cell survival. In Saccharomyces cerevisiae cells, regulation of the RP gene expression at the transcriptional, mature mRNA stability and translational levels during the response to osmotic stress has been reported. Reprogramming global protein synthesis upon osmotic shock includes the movement of ribosomes from RP transcripts to stress-induced mRNAs. Using tiling arrays, we show that osmotic stress yields a drop in the levels of RP pre-mRNAs in S. cerevisiae cells. An analysis of the tiling array data, together with transcription rates data, shows a poor correlation, indicating that the drop in the RP pre-mRNA levels is not merely a result of the lowered RP transcription rates. A kinetic study using quantitative RT-PCR confirmed the decrease in the levels of several RP-unspliced transcripts during the first 15 minutes of osmotic stress, which seems independent of MAP kinase Hog1. Moreover, we found that the mutations in the components of the nonsense-mediated mRNA decay (NMD), Upf1, Upf2, Upf3 or in exonuclease Xrn1, eliminate the osmotic stress-induced drop in RP pre-mRNAs. Altogether, our results indicate that the degradation of yeast RP unspliced transcripts by NMD increases during osmotic stress, and suggest that this might be another mechanism to control RP synthesis during the stress response

Checkpoints in a Yeast Differentiation Pathway Coordinate Signaling during Hyperosmotic Stress

All eukaryotes have the ability to detect and respond to environmental and hormonal signals. In many cases these signals evoke cellular changes that are incompatible and must therefore be orchestrated by the responding cell. In the yeast Saccharomyces cerevisiae, hyperosmotic stress and mating pheromones initiate signaling cascades that each terminate with a MAP kinase, Hog1 and Fus3, respectively. Despite sharing components, these pathways are initiated by distinct inputs and produce distinct cellular behaviors. To understand how these responses are coordinated, we monitored the pheromone response during hyperosmotic conditions. We show that hyperosmotic stress limits pheromone signaling in at least three ways. First, stress delays the expression of pheromone-induced genes. Second, stress promotes the phosphorylation of a protein kinase, Rck2, and thereby inhibits pheromone-induced protein translation. Third, stress promotes the phosphorylation of a shared pathway component, Ste50, and thereby dampens pheromone-induced MAPK activation. Whereas all three mechanisms are dependent on an increase in osmolarity, only the phosphorylation events require Hog1. These findings reveal how an environmental stress signal is able to postpone responsiveness to a competing differentiation signal, by acting on multiple pathway components, in a coordinated manner

The HOG Pathway Dictates the Short-Term Translational Response after Hyperosmotic Shock

In the global osmoshock translational response in yeast, some gene products were translationally mobilized without transcriptional up-regulation. Conversely, other transcriptionally up-regulated mRNAs were translationally inhibited. Analogous changes occurred on the protein level. These translational responses were strongly dependent on Hog1 and Rck2