Novel parent-of-origin-specific differentially methylated loci on chromosome 16

BACKGROUND: Congenital malformations associated with maternal uniparental disomy of chromosome 16, upd(16)mat, resemble those observed in newborns with the lethal developmental lung disease, alveolar capillary dysplasia with misalignment of pulmonary veins (ACDMPV). Interestingly, ACDMPV-causative deletions, involving FOXF1 or its lung-specific upstream enhancer at 16q24.1, arise almost exclusively on the maternally inherited chromosome 16. Given the phenotypic similarities between upd(16)mat and ACDMPV, together with parental allelic bias in ACDMPV, we hypothesized that there may be unknown imprinted loci mapping to chromosome 16 that become functionally unmasked by chromosomal structural variants. RESULTS: To identify parent-of-origin biased DNA methylation, we performed high-resolution bisulfite sequencing of chromosome 16 on peripheral blood and cultured skin fibroblasts from individuals with maternal or paternal upd(16) as well as lung tissue from patients with ACDMPV-causative 16q24.1 deletions and a normal control. We identified 22 differentially methylated regions (DMRs) with ≥ 5 consecutive CpG methylation sites and varying tissue-specificity, including the known DMRs associated with the established imprinted gene ZNF597 and DMRs supporting maternal methylation of PRR25, thought to be paternally expressed in lymphoblastoid cells. Lastly, we found evidence of paternal methylation on 16q24.1 near LINC01082 mapping to the FOXF1 enhancer. CONCLUSIONS: Using high-resolution bisulfite sequencing to evaluate DNA methylation across chromosome 16, we found evidence for novel candidate imprinted loci on chromosome 16 that would not be evident in array-based assays and could contribute to the birth defects observed in patients with upd(16)mat or in ACDMPV

Ethical issues in longitudinal studies: the case of ELSA-Brasil

Historicamente a discussão acerca da eticidade dos atos em pesquisas com seres humanos privilegiou os estudos experimentais, pelo maior potencial de danos aos sujeitos envolvidos. Todavia, os estudos observacionais também envolvem riscos e suscitam questões relevantes. Neste artigo pretende-se apresentar e discutir aspectos éticos do desenvolvimento do ELSA-Brasil, um estudo longitudinal e multicêntrico, com financiamento público, no qual os sujeitos da pesquisa e pesquisadores pertencem às mesmas instituições. São descritos os procedimentos adotados para atender às exigências e compromissos éticos e a casuística que orientou as ações segundo seus princípios norteadores (beneficência, autonomia e justiça social). São apresentados alguns problemas morais que exigiram ponderação sobre riscos e benefícios na confluência com os objetivos do estudo e comentam-se peculiaridades de um estudo longitudinal e seus potenciais benefícios.The debate about ethics in research with human beings has historically emphasized experimental studies because of their greater potential to harm the subjects involved. However, observational studies also include risks and relevant questions to be discussed. This article aims to present and discuss the ethical aspects involved in the implementation of ELSA-Brasil, a longitudinal multicenter study, with public funding, in which the research subjects and investigators are employees of the same institutions. The procedures adopted to meet the ethical requirements and commitments are described, as well as the casuistics that guided the actions according to their guiding principles (beneficence, autonomy and social justice). We present some moral problems that required consideration of risks and benefits at the confluence with the study's objectives, and we conclude with comments on the peculiarities and the potential benefits of a longitudinal study

Antiproliferative activity of flavonoids from croton Sphaerogynus baill. (euphorbiaceae)

Croton sphaerogynus is a shrub from the Atlantic Rain Forest in southeastern Brazil. A lyophilized crude EtOH extract from leaves of C. sphaerogynus, obtained by maceration at room temperature (seven days), was suspended in methanol and partitioned with hexane. The purified MeOH phase was fractionated over Sephadex LH-20 yielding five fractions (F1-F5) containing flavonoids, as characterized by HPLC-DAD and HPLC-MS analyses. The antiproliferative activity of the crude EtOH extract, MeOH and hexane phases, and fractions F1-F5 was evaluated on in vitro cell lines NCI-H460 (nonsmall cell lung), MCF-7 (breast cancer), and U251 (glioma). The MeOH phase showed activity (mean log GI(50) 0.54) higher than the hexane phase and EtOH extract (mean log GI(50) 1.13 and 1.19, resp.). F1 exhibited activity against NCI-H460 (nonsmall cell lung) (GI(50) 1.2 mu g/mL), which could be accounted for the presence of flavonoids and/or diterpenes. F4 showed moderate activity (mean log GI(50) 1.05), while F5 showed weak activity (mean log GI(50) 1.36). It is suggested that the antiproliferative activity of the crude EtOH extract and MeOH phase is accounted for a synergistic combination of flavonoids and diterpenes2015COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informação2012/10079-

A large CRISPR-induced bystander mutation causes immune dysregulation.

A persistent concern with CRISPR-Cas9 gene editing has been the potential to generate mutations at off-target genomic sites. While CRISPR-engineering mice to delete a ~360 bp intronic enhancer, here we discovered a founder line that had marked immune dysregulation caused by a 24 kb tandem duplication of the sequence adjacent to the on-target deletion. Our results suggest unintended repair of on-target genomic cuts can cause pathogenic bystander mutations that escape detection by routine targeted genotyping assays

Quantum numbers of the X(3872)X(3872) state and orbital angular momentum in its ρ0Jψ\rho^0 J\psi decay

Angular correlations in $B^+\to X(3872) K^+$ decays, with $X(3872)\to \rho^0 J/\psi$, $\rho^0\to\pi^+\pi^-$ and $J/\psi \to\mu^+\mu^-$, are used to measure orbital angular momentum contributions and to determine the $J^{PC}$ value of the $X(3872)$ meson. The data correspond to an integrated luminosity of 3.0 fb$^{-1}$ of proton-proton collisions collected with the LHCb detector. This determination, for the first time performed without assuming a value for the orbital angular momentum, confirms the quantum numbers to be $J^{PC}=1^{++}$. The $X(3872)$ is found to decay predominantly through S wave and an upper limit of $4\%$ at $95\%$ C.L. is set on the fraction of D wave.Comment: 16 pages, 4 figure

Observation of J/ψpJ/\psi p resonances consistent with pentaquark states in Λb0→J/ψK−p{\Lambda_b^0\to J/\psi K^-p} decays

Observations of exotic structures in the $J/\psi p$ channel, that we refer to as pentaquark-charmonium states, in $\Lambda_b^0\to J/\psi K^- p$ decays are presented. The data sample corresponds to an integrated luminosity of 3/fb acquired with the LHCb detector from 7 and 8 TeV pp collisions. An amplitude analysis is performed on the three-body final-state that reproduces the two-body mass and angular distributions. To obtain a satisfactory fit of the structures seen in the $J/\psi p$ mass spectrum, it is necessary to include two Breit-Wigner amplitudes that each describe a resonant state. The significance of each of these resonances is more than 9 standard deviations. One has a mass of $4380\pm 8\pm 29$ MeV and a width of $205\pm 18\pm 86$ MeV, while the second is narrower, with a mass of $4449.8\pm 1.7\pm 2.5$ MeV and a width of $39\pm 5\pm 19$ MeV. The preferred $J^P$ assignments are of opposite parity, with one state having spin 3/2 and the other 5/2.Comment: 48 pages, 18 figures including the supplementary material, v2 after referee's comments, now 19 figure

Precise measurements of the properties of the B-1(5721)(0,+) and B-2*(5747)(0,+) states and observation of B-+,B-0 pi(-,+) mass structures

Invariant mass distributions of B+π− and B0π+ combinations are investigated in order to study excited B mesons. The analysis is based on a data sample corresponding to 3.0 fb−1 of pp collision data, recorded by the LHCb detector at centre-of-mass energies of 7 and 8 TeV. Precise measurements of the masses and widths of the B1(5721)0,+ and B2(5747)0,+ states are reported. Clear enhancements, particularly prominent at high pion transverse momentum, are seen over background in the mass range 5850-6000 MeV in both B+π− and B0π+ combinations. The structures are consistent with the presence of four excited B mesons, labelled BJ (5840)0,+ and BJ (5960)0,+, whose masses and widths are obtained under different hypotheses for their quantum numbers

Atuação da comissão de farmácia e terapêutica em um hospital de ensino

The hospital activities are characterized by a highly dynamism as a result of new health technologies such as medicines. A hospital due its characteristics of teaching, research and high complexity care, has the highest concentration of different types of health technologies. The Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto – USP (HCFMRP-USP) is a hospital, with proven quality, inserted in the SUS as a tertiary/quaternary referral and has the Pharmaceutical Services Division (DAF) for development of actions of health care. To aid resource management, selection and standardizationof drugs, DAF adopted the strategy of Pharmacy and Therapeutics Committee (CFT). The CFT is a collegial, consultative and deliberative body, established by the World Health Organization as a strategy tool to monitor and promote the quality in the use of medicine, but studies of CFTs are incipient in Brazil. Thus, this study aims to present the CFT of HCFMRP–USP. Objectives: To introduce the composition, responsibilities and working methods of CFT, as well as a critical analysis of its current operation. Methods: A descriptive study aimed to describe the current functioning of the CFT of HCFMRP-USP was performed. Ordinances, internal regulations were surveyed and a bibliographic review of the CFT was performed. To the critical analysis of the current operating, was selected by the committee from the standard one that would fit classification as belonging to “A” and “V” items after the crossing of the curves ABC and VEN, whose selected item was the medicine Sevoflurane. Results: The CFT was established in 2010 to replace the defunct standardization committee. Since then, the CFT examined 134 requests and 41 of these were standardized. The Sevoflurane drug was incorporated into the HCFMRP-USP in 2010 and, starting that year, there was a gradual increase in the consumption of the same. However, after analyzing the requirements of the drug in 2012, it was observed that the dispensation of Sevoflurane does not follow the specifications of the patient profile as established in the protocol established at the time of standardization. Conclusions: We concluded that the implementation of CFT was a strategy that provided a rational standardization. However, it is observed that there is no control of dispensing and use of the product according to the protocol established at the time of standardization. We emphasize that control the use of Sevoflurane is not responsibility of the CFT and this assignment should be delegated to the responsible technical area.As atividades hospitalares caracterizam-se por um acentuado dinamismo em consequência do surgimento de novas tecnologias em saúde, tais como medicamentos. Uma unidade hospitalar, devido suas características de ensino, pesquisa e atendimentos de alta complexidade, possui maior concentração de diferentes tipos de tecnologias em saúde. O Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto –USP (HCFMRP-USP) é uma instituição hospitalar, de qualidade comprovada, inserido no SUS como referência terciária/quaternária e que conta com a Divisão de Assistência Farmacêutica (DAF) para desenvolvimento das ações de atenção a saúde. A DAF para auxílio da gestão de recursos, seleção e padronização de medicamentos adotou como estratégia a Comissão de Farmácia e Terapêutica (CFT). A CFT é uma instância colegiada, de caráter consultivo e deliberativo, estabelecida pela Organização Mundial de Saúde como ferramenta de estratégia para monitorar e promover a qualidade no uso do medicamento, porém estudos que sobre a atuação das CFTs no Brasil são incipientes. Desta forma, este estudo pretende apresentar a CFT do HCFMRP-USP. Objetivos: apresentar a composição, atribuições e metodologia de trabalho da CFT, bem como desenvolver uma análise crítica de seu atual funcionamento. Metodologia: Foi realizado estudo descritivo e exploratório com o objetivo de descrever o atual funcionamento da CFT do HCFMRP-USP. Foram buscadas portarias, regulamentações internas e foi realizada revisão bibliográfica sobre a CFT. Para Análise crítica do atual funcionamento, foi selecionado dentre os itens padronizados pela comissão aquele que se enquadrasse como pertencente a classificação A e V, após o cruzamento das curvas ABC e VEN, cujo item selecionado foi o medicamento Sevoflurano. Resultados: A CFT foi instituída no ano de 2010 em substituição a extinta comissão de padronização. Desde então, a CFT analisou 134 solicitações e destas 41 foram padronizadas. O medicamento sevoflurano foi incorporado no HCFMRP-USP em 2010 e, a partir deste ano, observa-se um aumento gradativo do consumo do mesmo. Entretanto, após análise das prescrições do referido medicamento no ano de 2012, foi observado que a dispensação do sevoflurano não segue as especificações do perfil de pacientes conforme estabelecido no protocolo instituído no momento da padronização. Conclusões: Portanto, concluímos que a implantação da CFT foi uma estratégia que proporcionou a padronização racional. Entretanto, observa-se que não há controle da dispensação e utilização do medicamento de acordo com o protocolo estabelecido no momento da padronização. Salientamos que o controle do uso do Sevoflurano não é atribuição da CFT devendo esta atribuição ser delegada à área técnica responsável