Desenvolvimento de abordagens de espectroscopia de fibra ótica para a análise em alto débito de estirpes de Saccharomyces cerevisiae

Dissertação de mestrado em Genética MolecularO objetivo deste trabalho consistiu na implementação de um teste espectrométrico miniaturizado e de alto rendimento para a análise de estirpes de Saccharomyces cerevisiae da coleção do Centro de Biologia Molecular e Ambiental da Universidade do Minho. Para atingir esse objetivo foi necessário desenvolver um suporte para microplacas de 96 poços onde se realizaram as medições, no sentido de encontrar as melhores condições de crescimento para as estirpes e a posterior aquisição dos espectros. Os espectros foram adquiridos na zona da luz ultravioletavisível- infravermelha próximo de ondas curtas (200 – 1100 nm). As estirpes utilizadas foram inoculadas em meio YPD agarizado numa microplaca de 96 poços. Os espectros obtidos foram submetidos a pré-tratamentos e posteriormente a análise de componentes principais relevantes e a análise discriminante por mínimos quadrados parciais, quando indicado. Ensaios preliminares de otimização sugeriram a necessidade de ter em atenção determinados cuidados para minimizar os erros provenientes do material utilizado e que podiam interferir com a análise espectral das estirpes. Foi demonstrada a possibilidade de distinguir 11 estirpes da levedura S. cerevisiae, provenientes de diferentes localizações geográficas e origens de isolamento, bem como de 9 isolados isogénicos da estirpe comercial de vinificação Zymaflore VL1 (Lallemand), desde que se consideram condições experimentais muito padronizados como, por exemplo, a obtenção de espectros que foram obtidos na mesma microplaca. Com esta abordagem foi alcançada uma boa reprodutibilidade de réplicas de crescimentos e medições espectrométricas. No entanto, não foi obtida uma boa reprodutibilidade entre microplacas contendo as mesmas estirpes. Verificou-se também que a semelhança espectral não está relacionada com a localização geográfica e/ou a origem do isolamento das estirpes. As modificações espectrais foram também registadas durante o crescimento das estirpes nos poços das microplacas. As análises posteriores distinguiram as diferentes fases de crescimento, mas para o tempo de incubação entre 42 h e 76 h não se verificaram diferenças espectrais significativas. Em todas as experiências anteriores, a identidade de cada estirpe analisada era conhecida. Para validar este método espectroscópico foi necessário verificar a possibilidade de se conseguir distinguir isolados da estirpe S. cerevisiae sem conhecimento prévio da sua identidade. Os resultados das análises espectrais obtidos foram comparados com os de perfis moleculares (análise interdelta) destes isolados e não se observou uma correspondência entre o número de estirpes identificado por cada um dos métodos. Por este motivo, trata-se de uma técnica que não poderá ser utilizada para distinguir estirpes cuja identidade seja desconhecida. Sendo uma técnica no início do seu desenvolvimento, verificou-se a ocorrência de vários problemas sistemáticos nas experiências realizadas, inerentes ao material utilizado e devido à variação na composição das microplacas e do meio de cultura. Contudo, foram apresentadas soluções para a obtenção de resultados mais fiáveis e reprodutíveis no futuro.The aim of this work was to develop a high-throughput spectrometric assay for the analysis of Saccharomyces cerevisiae strains from the collection of the Centre for Molecular Biology and Environment, University of Minho. To achieve this goal it was necessary to develop a support for 96-well microplates for small-scale spectra acquisition and for the development of the best growth conditions. The spectra were acquired in the region of ultraviolet-visible-near infrared short wave light (200 - 1100 nm). The strains used were inoculated in wells of a 96-well microplate containing YPD agar. The spectra obtained were subjected to pre-treatments and subsequently to relevant principal components analysis and the discriminant analysis by partial least squares, whenever indicated. Preliminary optimization tests suggested the need to consider several aspects (such as the material used), that could interfere with the spectral analysis of the S. cerevisiae strains. It was possible to distinguish between 11 S. cerevisiae strains from different geographical locations and sources of isolation and 9 isogenic isolates of the commercial winemaking strain Zymaflore VL1 (Lallemand), that were re-isolated from vineyards when the obtained spectra derived from a single microplate. The distinction of strains occurred with a good reproducibility of the four replicates included in each microplate. However, a lack of reproducibility occurred between spectra obtained from different microplates that contained the same strains. It was also found that the spectral similarity of strains did not correlate with the geographic localisation and the source of isolation. Spectral analyses distinguished different stages of exponential cellular growth, whereas no significant spectral differences were recorded between 42 h and 76 h of growth. In all previous experiments, the identity of each strain analysed was known. For further validation of this spectroscopic method, we evaluated the ability to distinguish among several S. cerevisiae isolates that were previously characterized by interdelta analysis. The comparison of results from spectral analysis with interdelta profiles revealed the number of strains identified by each method was different. We herein developed a first exploratory approach for the high throughput analysis of S. cerevisiae strains by fiber optics spectroscopy. Several systemic problems, inherent to the materials used, such as variation in the composition of the microplate and the culture medium still need to be solved and we present solutions for the achievement of more reliable and reproducible results in the future

OUTCOMES OF ACCIDENTS AT WORK WITH EXPOSURE

Introduction: In the labour process there are several occupational hazard risks that present themselves invisibly to workers. Occupational exposure to biological material via sharps is high among health workers. Biosafety is an integral part of safety and occupational hygiene. The risk of contamination by biological material is inherent in anything exposed to it, but it varies according to the activities undertaken by workers. Objective: To characterise the outcomes of accidents at work involving the exposure of workers to biological agents, in the ABC Paulista region. Methods: This was a descriptive study using a convenience sample from three municipalities in the Grande ABC region: Santo André, São Bernardo do Campo and São Caetano do Sul. The data was sourced from 47 records of reported accidents with biological material in the Greater ABC region in the year 2012. The data was processed and tabulated in the statistical program SPSS 19. We used notification records and obtained the profiles of the injured workers and characteristics of the accident, according to gender, age, county of residence, profession, occupation, type of exposure, material involved, the situation of the agent causing the accident, use of personal protective equipment (PPE) equipment, and communication about the work accident (CAT). Results: The incidence of accidents with sharps was related to the frequent handling of these objects and the behaviour of professionals who use practices that pose the risk of needlestick injuries, such as the improper disposal of sharps. Conclusion: The main cause of biological accidents was sharps. Females (70.2 %) and nursing staff (63.8 %) were more likely to be involved in accidents

Segurança contra incêndio em unidades básicas de saúde

Nowadays it is widely recognized the demand of a modernization of the Government and his administrative entities. According to the legislation referent to fire prevention the Firefighters were the most proactive entity concerning the creation of legislation based on the tragic incidents and the experience acquired in daily routines. A fire can occur in any situation whether exist or not human activities and Basic Health Units are not free from this event and unfortunately a real possibility of this happening due to elevated fire charge. The purpose of this study was to evaluate the level of knowledge and that employees have in relation to Fire Safety theme and confront the findings with the needs required by law due to the non-mandatory fire brigade constitution in Basic Health Units with ground floor and inferior to 750 m² whose requisites fulfill the vast majority of buildings. The research method is qualitative observational and descriptive by convenience sample in nine Basic Health Units in city council Santo André,(São Paulo, Brazil) with the aim to enrich the health professionals knowledge about fire prevention was also evaluated in this case study.Hoje em dia reconhece-se a necessidade de uma modernização dos Estados e respectivos aparelhos administrativos. No contexto normativo e referente à prevenção de incêndios, o Corpo de Bombeiros foi à entidade mais atuante na criação de legislações e normas, baseando-se nos trágicos acontecimentos e na experiência adquirida no atendimento diário de ocorrências. O incêndio pode atacar qualquer local onde existam ou não atividades humanas sendo que os estabelecimentos assistenciais de saúde (EAS) não estão isentos deste infortúnio e infelizmente existe a possibilidade real de que possam acontecer novas tragédias relacionadas com incêndios dado que a carga de incêndio. O objectivo deste estudo foi o de avaliar o nível de conhecimento que os funcionários de Unidades Básicas de Saúde possuem em relação à temática Segurança contra Incêndio e confrontar os resultados com o exigido legislação vigente confrontando os achados com o Decreto Estadual 5.6819/11 devido à não obrigatoriedade de constituição de brigada de incêndio nas unidades básicas de saúde, com menos de 750 m², piso térreo, constituindo a grande maioria das edificações do sistema de Saúde Brasileiro. O método de investigação é um qualitativo observacional e descritivo por amostra de conveniência em noveUnidades Básicas de Saúde no município de Santo André, São Paulo, Brasil, com o objetivo de aprofundar o nível de conhecimento que os funcionários possuem em relação à temática Segurança contra Incêndio

Socio-Environmental and Hematological Profile of Landfill Residents (Sao Jorge Landfill-Sao Paulo, Brazil)

We are experiencing an unprecedented urbanization process that, alongside physical, social and economic developments, has been having a significant impact on a population's health. Due to the increase in pollution, violence and poverty, our modern cities no longer ensure a good quality of life so they become unhealthy environments. This study aims to assess the effect of social, environmental and economic factors on the hematologic profile of residents of Santo Andre's landfill. In particular, we will assess the effect of social, economic, and environmental factors on current and potential disease markers obtained from hematological tests. The research method is the observational type, from a retrospective cohort, and by convenience sampling in Santo Andre in the Greater ABC (municipalities of Santo Andre, Sao Bernardo do Campo and Sao Caetano do Sul, southeast part of the Greater Sao Paulo Metropolitan Area, Brazil). The study determined a socio-environmental profile and the hematologic diseases screening related to a close location to the landfill. The disease manifests itself within a broad spectrum of symptoms that causes changes in blood count parameters. The objective of this work is to show that there is an association between social, environmental and economic factors and a variety of serious disease outcomes that may be detected from blood screening. A causal study of the effect of living near the landfill on these disease outcomes would be a very expensive and time-consuming study. This work we believe is sufficient for public health officials to consider policy and attempt remediation of the effects of living near a landfill.ABC Medical SchoolSchool of Health Technology Coimbra-IPC-Polytechnic Institute of Coimbra-PortugalDepartment of Environmental Health and Public HealthSEMASA-Sanitation and Insurance CentralABC MedSch, Environm Hlth Management Dept, BR-09060650 Santo Andre, SP, BrazilUniv Fed Sao Paulo, Inst Environm Chem & Pharmaceut Sci, Dept Biol Sci, BR-04060650 Diadema, SP, BrazilCoimbra Hlth Sch, EsTesC, Dept Environm Hlth Sci, P-3046854 Coimbra, PortugalBiological Sciences Department, Institute of Environmental, Chemistry and Pharmaceutical Sciences, Universidade Federal de São Paulo (UNIFESP), Diadema 04060-650, SP, BrazilWeb of Scienc

A continuous integration and web framework in support of the ATLAS publication process

The ATLAS collaboration defines methods, establishes procedures, and organises advisory groups to manage the publication processes of scientific papers, conference papers, and public notes. All stages are managed through web systems, computing programs, and tools that are designed and developed by the collaboration. A framework called FENCE is integrated into the CERN GitLab software repository, to automatically configure workspaces where each analysis can be documented by the analysis team and managed by the relevant coordinators. Continuous integration is used to guide the writers in applying consistent and correct formatting when preparing papers to be submitted to scientific journals. Additional software assures the correctness of other aspects of each paper, such as the lists of collaboration authors, funding agencies, and foundations. The framework and the workflow therein provide automatic and easy support to the researchers and facilitates each phase of the publication process, allowing authors to focus on the article contents. The framework and its integration with the most up to date and efficient tools has consequently provided a more professional and efficient automatized work environment to the whole collaboration.ATLAS Collaboration for the support provided to achieve the results described in this paper. We are grateful to ATLAS collaborators who provided invaluable comments and input to the paper and the framework it presents. Special acknowledgements go to Marzio Nessi for helping initiate the Glance project in ATLAS and for supporting its development, and to Kathy– 20 –Pommes for supervising the Glance team at CERNinfo:eu-repo/semantics/publishedVersio

SARS-CoV-2 introductions and early dynamics of the epidemic in Portugal

Genomic surveillance of SARS-CoV-2 in Portugal was rapidly implemented by the National Institute of Health in the early stages of the COVID-19 epidemic, in collaboration with more than 50 laboratories distributed nationwide. Methods By applying recent phylodynamic models that allow integration of individual-based travel history, we reconstructed and characterized the spatio-temporal dynamics of SARSCoV-2 introductions and early dissemination in Portugal. Results We detected at least 277 independent SARS-CoV-2 introductions, mostly from European countries (namely the United Kingdom, Spain, France, Italy, and Switzerland), which were consistent with the countries with the highest connectivity with Portugal. Although most introductions were estimated to have occurred during early March 2020, it is likely that SARS-CoV-2 was silently circulating in Portugal throughout February, before the first cases were confirmed. Conclusions Here we conclude that the earlier implementation of measures could have minimized the number of introductions and subsequent virus expansion in Portugal. This study lays the foundation for genomic epidemiology of SARS-CoV-2 in Portugal, and highlights the need for systematic and geographically-representative genomic surveillance.We gratefully acknowledge to Sara Hill and Nuno Faria (University of Oxford) and Joshua Quick and Nick Loman (University of Birmingham) for kindly providing us with the initial sets of Artic Network primers for NGS; Rafael Mamede (MRamirez team, IMM, Lisbon) for developing and sharing a bioinformatics script for sequence curation (https://github.com/rfm-targa/BioinfUtils); Philippe Lemey (KU Leuven) for providing guidance on the implementation of the phylodynamic models; Joshua L. Cherry (National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health) for providing guidance with the subsampling strategies; and all authors, originating and submitting laboratories who have contributed genome data on GISAID (https://www.gisaid.org/) on which part of this research is based. The opinions expressed in this article are those of the authors and do not reflect the view of the National Institutes of Health, the Department of Health and Human Services, or the United States government. This study is co-funded by Fundação para a Ciência e Tecnologia and Agência de Investigação Clínica e Inovação Biomédica (234_596874175) on behalf of the Research 4 COVID-19 call. Some infrastructural resources used in this study come from the GenomePT project (POCI-01-0145-FEDER-022184), supported by COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation (POCI), Lisboa Portugal Regional Operational Programme (Lisboa2020), Algarve Portugal Regional Operational Programme (CRESC Algarve2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), and by Fundação para a Ciência e a Tecnologia (FCT).info:eu-repo/semantics/publishedVersio

Juxtaposing BTE and ATE – on the role of the European insurance industry in funding civil litigation

One of the ways in which legal services are financed, and indeed shaped, is through private insurance arrangement. Two contrasting types of legal expenses insurance contracts (LEI) seem to dominate in Europe: before the event (BTE) and after the event (ATE) legal expenses insurance. Notwithstanding institutional differences between different legal systems, BTE and ATE insurance arrangements may be instrumental if government policy is geared towards strengthening a market-oriented system of financing access to justice for individuals and business. At the same time, emphasizing the role of a private industry as a keeper of the gates to justice raises issues of accountability and transparency, not readily reconcilable with demands of competition. Moreover, multiple actors (clients, lawyers, courts, insurers) are involved, causing behavioural dynamics which are not easily predicted or influenced. Against this background, this paper looks into BTE and ATE arrangements by analysing the particularities of BTE and ATE arrangements currently available in some European jurisdictions and by painting a picture of their respective markets and legal contexts. This allows for some reflection on the performance of BTE and ATE providers as both financiers and keepers. Two issues emerge from the analysis that are worthy of some further reflection. Firstly, there is the problematic long-term sustainability of some ATE products. Secondly, the challenges faced by policymakers that would like to nudge consumers into voluntarily taking out BTE LEI

Penilaian Kinerja Keuangan Koperasi di Kabupaten Pelalawan

This paper describe development and financial performance of cooperative in District Pelalawan among 2007 - 2008. Studies on primary and secondary cooperative in 12 sub-districts. Method in this stady use performance measuring of productivity, efficiency, growth, liquidity, and solvability of cooperative. Productivity of cooperative in Pelalawan was highly but efficiency still low. Profit and income were highly, even liquidity of cooperative very high, and solvability was good

Differential cross section measurements for the production of a W boson in association with jets in proton–proton collisions at √s = 7 TeV

Measurements are reported of differential cross sections for the production of a W boson, which decays into a muon and a neutrino, in association with jets, as a function of several variables, including the transverse momenta (pT) and pseudorapidities of the four leading jets, the scalar sum of jet transverse momenta (HT), and the difference in azimuthal angle between the directions of each jet and the muon. The data sample of pp collisions at a centre-of-mass energy of 7 TeV was collected with the CMS detector at the LHC and corresponds to an integrated luminosity of 5.0 fb[superscript −1]. The measured cross sections are compared to predictions from Monte Carlo generators, MadGraph + pythia and sherpa, and to next-to-leading-order calculations from BlackHat + sherpa. The differential cross sections are found to be in agreement with the predictions, apart from the pT distributions of the leading jets at high pT values, the distributions of the HT at high-HT and low jet multiplicity, and the distribution of the difference in azimuthal angle between the leading jet and the muon at low values.United States. Dept. of EnergyNational Science Foundation (U.S.)Alfred P. Sloan Foundatio