A comprehensive integrative approach to investigate factors associated with preterm birth, related perinatal outcomes and its prediction using metabolomic markers

Orientador: José Guilherme CecattiTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: Introdução: O parto prematuro é uma das principais causas de morbimortalidade perinatal, neonatal e de crianças até 5 anos de idade e suas causas e fisiopatologia ainda são pouco conhecidas. Identificar quais são as mulheres de maior risco e desenvolver modelos de predição é ainda um grande desafio, potencialmente impactando nas medidas preventivas. Objetivo: Desenvolver uma abordagem abrangente com diferentes estudos e produtos relacionados aos fatores clínicos e epidemiológicos associados ao parto prematuro, seus preditores metabolômicos e respectivos desfechos perinatais. Métodos: Diferentes projetos de pesquisa e métodos foram utilizados, incluindo: duas análises secundárias de um estudo multicêntrico de corte transversal avaliando a associação do índice de massa corpórea (IMC), o ganho de peso gestacional por semana e fenótipos maternos com a ocorrência de prematuridade e desfechos maternos e perinatais; uma revisão narrativa sobre ciência ômica aplicada na área de saúde materna e perinatal, com enfoque na metabolômica; uma revisão sistemática e seu respectivo artigo de protocolo sobre a performance da metabolômica em predizer prematuridade espontânea em mulheres assintomáticas; dois artigos abordando o desenvolvimento do método e dos procedimentos técnicos para um estudo multicêntrico prospectivo para investigar parto prematuro; um estudo caso-controle aninhado a uma coorte multicêntrica internacional para identificar preditores clínicos e metabolômicos para prematuridade espontânea; dois artigos originais abordando a incidência, fatores clínicos e epidemiológicos e os desfechos maternos e perinatais associados ao parto prematuro em uma coorte multicêntrica no Brasil com gestantes nulíparas de baixo risco. Resultados: Nas análises secundárias do EMIP, observou-se que independente do IMC inicial, quanto maior o ganho de peso materno, maior a probabilidade para todos os subtipos de prematuridade, exceto para prematuridade espontânea em mulheres com IMC normal ou sobrepeso. Foram identificados três clusters de mulheres com parto prematuro, sendo um caracterizado principalmente por mulheres sem nenhuma das condições de risco, o segundo por mulheres com várias condições (cluster misto) e o terceiro por mulheres que tiveram pré-eclâmpsia, eclâmpsia, síndrome HELLP e/ou restrição de crescimento fetal. A revisão narrativa aborda os métodos e o embasamento teórico das ciências ômicas, como a genômica, transcriptômica, proteômica e metabolômica, dando enfoque especial à aplicação dessa última técnica na área de saúde materna e perinatal. A identificação e validação de marcadores pode auxiliar na predição e também no entendimento da fisiopatologia de doenças complexas como a prematuridade. A técnica de metabolômica identificou mais de 140 metabólitos nas amostras de soro de gestantes nulíparas e três destes foram significativamente associados com parto prematuro espontâneo nas amostras de Cork, Irlanda. Modelos preditores usando marcadores clínicos e metabolômicos mostraram uma área sob a curva ROC de 0,73 e 0,85 para parto prematuro abaixo de 37 e 34 semanas, respectivamente. Conclusão: O ganho de peso gestacional, um fator modificável, mostrou diferentes associações com a probabilidade de parto prematuro, a depender do IMC inicial. Possíveis investigações de risco e de prevenção devem considerar essa evidência. A utilização de critérios clínicos no rastreamento e predição do parto prematuro ainda mostra limitações. A análise por cluster, por exemplo, mostrou que um número considerável não possui nenhuma das condições pré-definidas como potencialmente associadas ao parto prematuro. A aplicação de estudos da ciência Ômica parece ser uma abordagem adequada para a identificação da etiologia e de marcadores para predição de complicações maternas e perinatais, embora ainda necessitem de sucessivas validações e evidência de reprodutibilidade. O desenvolvimento, implementação e coordenação de um estudo multicêntrico para estudar preditores e fatores associados ao parto prematuro requer recursos humanos qualificados, infraestrutura para pesquisa adequada, comprometimento institucional e envolvimento de agências de fomento e desenvolvimento de pesquisa. O modelo preditor para parto prematuro espontâneo em mulheres nulíparas mostra resultados de boa performance, entretanto requer futuras validações antes de qualquer uso clínico. É provável que os metabólitos que compõem o modelo não sejam identificados da mesma forma em outras populaçõesAbstract: Introduction: Preterm birth is the leading cause of perinatal, neonatal and under-5 year¿s morbidity and mortality. Identifying women at higher risk and developing prediction models remains a great challenge, potentially affecting preventive interventions. Objectives: To develop a comprehensive approach including diverse study designs to investigate clinical and epidemiological risk factors associated with preterm birth, its metabolomics predictors and respective perinatal outcomes. Methods: Different projects and methods were applied in this thesis, including: two secondary analysis of a multicentre cross-sectional with a nested case-control study addressing the association of maternal body mass index (BMI), gestational weight gain per week and phenotypes with the occurrence of preterm birth and maternal and perinatal outcomes; an integrative review about omics sciences applied to maternal and perinatal health, focusing on metabolomics; a systematic review and respective protocol investigating the performance of metabolomics to predict spontaneous preterm birth (sPTB) in asymptomatic women; two articles describing the methods, clinical protocol, technical procedures for the development and implementation of a multicentre prospective cohort study to investigate preterm birth and other maternal and perinatal complications; a nested case-control from a multicentre international cohort to identify clinical and metabolomics predictors for sPTB; two articles addressing incidence, clinical and epidemiological risk factors and maternal and perinatal outcomes associated with sPTB in a Brazilian multicentre cohort of low-risk nulliparous pregnant women. Results: According to the EMIP secondary analyses, the greater the rate of weight gain, the higher the predicted probability for all preterm birth subtypes regardless the initial BMI, except in normal BMI or overweight women and sPTB. Three clusters of women with preterm birth were identified; cluster one of women without any pre-defined conditions, cluster two with mixed conditions and cluster three with women who had preeclampsia, eclampsia, HELLP syndrome and/or fetal growth restriction. Maternal and perinatal outcomes did not differ between clusters. An integrative review addressed Omis Science's methods and theoretical background, as genomics, transcriptomics, proteomics and metabolomics, focusing on the application on maternal and perinatal health. Metabolomics approach has been applied to better understand the pathophysiology and to identify and validate predictors for complex diseases as preterm birth. Metabolomics technique identified more than 140 metabolites in serum samples of nulliparous pregnant women and three of them were significantly associated with sPTB in samples from Cork, Ireland. Predictive models associating metabolites and clinical markers showed an area under ROC curve of 0.73 and 0.85 for sPTB below 37 and 34 weeks, respectively. Conclusion: Gestational weight gain, a modifiable factor, showed to have different associations with the predicted probability for preterm birth, depending on the initial BMI. The use of clinical criteria in the screening of preterm birth still shows limited performance. Cluster analysis, for instance, showed that a substantial number of women does not present the predefined potential conditions associated with preterm birth. Omics science studies might be a reasonable approach to investigate the aetiology and predictive markers for maternal and perinatal complications. Metabolomic studies addressing the prediction for sPTB, preeclampsia, gestational diabetes mellitus and fetal growth restriction show promising findings, although they still require repeated validations and reproducibility. The development, implementation and management of a multicenter study to investigate factors associated with sPTB requires qualified human resources, adequate infrastructure, institutional commitment and the involvement of funding and research agencies. The predictive model for sPTB in nulliparous women showed a good performance, although further validation is required before clinical application. Possibly, reproducibility of the predictive model is limited, once metabolites comprising the model were only identified in one of the subsetDoutoradoSaúde Materna e PerinatalDoutor em Ciências da SaúdeCAPE

Análise da influência dos métodos de aplicação do fluido de corte na rugosidade e na força geradas no torneamento do aço inoxidável AISI 420C

Visando reduzir os impactos negativos gerados pelo uso dos fluidos de corte sem reduzir a qualidade dos produtos e processos de usinagem, este estudo tem o objetivo de avaliar a influência de diferentes métodos de aplicação dos fluidos de corte, incluindo corte a seco, no torneamento do aço AISI 420C através da análise das componentes da força de usinagem e da rugosidade das superfícies usinadas. Para isto, um experimento fatorial completo foi realizado utilizando como fatores o método lubrirrefrigerante em quatro níveis, corte a seco, com fluido em abundância, com Mínima quantidade de lubrificante (MQL) e com Mínima quantidade de lubrificante resfriado (MQCL), e a velocidade de corte em dois níveis (200 e 300 m.min-1). Os resultados indicaram que o corte a seco, MQL ou MQCL são alternativas viáveis ao fluido em abundância, pois os parâmetros de saída foram pouco afetados pelos fatores analisados.Aiming to reduce the negative issues associatedto the use of cutting fluids without reducing the quality of manufactured products and productivity, this study aims to accessing the influence of different cutting fluid application techniques, including dry machining, in the turning of AISI 420C stainless steel through the analysis of the machining force components and the roughness of the machined surfaces. For this, a complete factorial experiment was performed, were the lubricooling methods were evaluated in four levels, dry cutting, flood, Minimum Quantity Lubrication (MQL) and Minimum Quantity Cooling Lubrication (MQCL), and the cutting speed in two levels (200 and 300 m.min-1). The results showed low influence of the cutting fluid application technique over the response variables, thus indicating that dry cutting, MQL or MQCL are viable alternatives to flood lubrication when these response variables are considered

O PROGRAMA RESIDÊNCIA PEDAGÓGICA E AS AULAS DE QUÍMICA NA PANDEMIA

The Pedagogical Residency is a program offered by CAPES, which is part of a public policy aimed at encouraging and valuing teacher education. The program aims to enhance the training of undergraduate students in teaching programs, actively exercising the relationship between theory and the professional teaching practice. Through a qualitative research methodology, such as a case study, a questionnaire was administered to the supervising teachers participating in the Pedagogical Residency, in order to understand how the state and schools provided training and support for online classes during the pandemic, as well as to investigate how Chemistry classes were conducted remotely. It was observed that the state and schools did not provide adequate support and training to teachers and students during remote classes, and the Chemistry classes had few methodological variations. In this context, students in the Chemistry Teaching program proposed activities with diverse teaching strategies, such as experiments and playful activities, to be used during online classes, aiding teachers in the complex process of teaching during the pandemic.La Residencia Pedagógica es un programa ofrecido por la CAPES, que forma parte de una política pública destinada a fomentar y valorar la formación de profesores. El programa tiene como objetivo mejorar la formación de los estudiantes de los cursos de licenciatura, ejercitando de manera activa la relación entre la teoría y la práctica profesional docente. A través de una metodología cualitativa de investigación, como el estudio de casos, se aplicó un cuestionario a los profesores preceptores que participaron en la Residencia Pedagógica, con el fin de conocer cómo el Estado y las escuelas proporcionaron capacitación y apoyo para las clases en línea durante la pandemia, y también para investigar cómo se llevaron a cabo las clases de Química de forma remota. Se observó que ni el Estado ni las escuelas brindaron un adecuado apoyo ni formación a los profesores y estudiantes durante las clases en línea, y que las clases de Química tenían escasas variaciones metodológicas. En este sentido, los residentes del curso de Licenciatura en Química propusieron actividades con diversas estrategias de enseñanza, como experimentos y actividades lúdicas, para ser utilizadas durante las clases en línea, asistiendo a los profesores en el complejo proceso de enseñanza durante la pandemia.A Residência Pedagógica é um programa ofertado pela CAPES, que faz parte de uma política pública que visa o incentivo e a valorização na formação de professores. O programa apresenta o objetivo de aperfeiçoar a formação dos discentes dos cursos de licenciatura, exercitando ativamente a relação entre a teoria e prática profissional docente. Por meio de uma metodologia qualitativa de pesquisa, do tipo estudo de caso, aplicou-se um questionário para os professores preceptores participantes da Residência Pedagógica, a fim de saber como o Estado e as escolas ofereceram treinamentos e suportes para as aulas online, durante a pandemia, bem como investigar como ocorreram as aulas remotas de Química. Notou-se que o Estado e as escolas não ofereceram suporte e treinamento adequado aos professores e alunos durante as aulas remotas, e as aulas de Química tinham poucas variações metodológicas. E nesse sentido, residentes do curso de Licenciatura em Química propuseram atividades com estratégias de ensino variadas, como experimentos e atividades lúdicas, para serem utilizados durante as aulas online, auxiliando os professores no complexo processo de regência durante a pandemia

Changes in the quality of cause-of-death statistics in Brazil : garbage codes among registered deaths in 1996–2016

Background: Registered causes in vital statistics classified as garbage codes (GC) are considered indicators of quality of cause-of-death data. Our aim was to describe temporal changes in this quality in Brazil, and the leading GCs according to levels assembled for the Global Burden of Disease (GBD) study. We also assessed socioeconomic differences in the burden of different levels of GCs at a regional level. Methods: We extracted data from the Brazilian Mortality Information System from 1996 to 2016. All three- and fourdigit ICD-10 codes considered GC were selected and classified into four categories, according to the GBD study proposal. GC levels 1 and 2 are the most damaging unusable codes, or major GCs. Proportionate distribution of deaths by GC levels according selected variables were performed. Age-standardized mortality rates after correction of underreporting of deaths were calculated to investigate temporal relationships as was the linear association adjusted for completeness between GC rates in states and the Sociodemographic Index (SDI) from the GBD study, for 1996–2005 and 2006–2016. We classified Brazilian states into three classes of development by applying tertiles cutoffs in the SDI state-level estimates. Results: Age-standardized mortality rates due to GCs in Brazil decreased from 1996 to 2016, particularly level 1 GCs. The most important GC groups were ill-defined causes (level 1) in 1996, and pneumonia unspecified (level 4) in 2016. At state level, there was a significant inverse association between SDI and the rate of level 1–2 GCs in 1996– 2005, but both SDI and completeness had a non-expected significant direct association with levels 3–4. In 2006– 2016, states with higher SDIs tended to have lower rates of all types of GCs. Mortality rates due to major GCs decreased in all three SDI classes in 1996–2016, but GC levels 3–4 decreased only in the high SDI category. States classified in the low or medium SDI groups were responsible for the most important decline of major GCs. Conclusion: Occurrence of major GCs are associated with socioeconomic determinants over time in Brazil. Their reduction with decreasing disparity in rates between socioeconomic groups indicates progress in reducing inequalities and strengthening cause-of-death statistics in the country

Efficacy of albendazole and moxidectin and resistance to ivermectin against Libyostrongylus douglassii and Libyostrongylus dentatus in ostriches

AbstractAnthelmintic resistance has emerged globally as a problem amongst nematode of livestock and has been particularly well documented in equine and small ruminants. There are no studies regarding the efficacy of anthelmintics against the hematophagous nematodes in ostriches, Libyostrongylus dentatus; and just a few on L. douglassii. Here the efficacy of albendazole, ivermectin and moxidectin were evaluated against these two species in an ostrich farm in Minas Gerais state, Brazil. The feces were collected on the day of treatment and after 13days of an oral dose of albendazole (6mg/kg), or an injected dose (0.2mg/kg) of ivermectin or moxidectin. The fecal egg count reduction test and coprocultures were performed to determine possible resistance against the drugs used. An efficacy of 60% was found for ivermectin, while albendazole and moxidectin were 100% effective. Both worm species appeared to have reduced sensitivity to ivermectin

Association between cervical length and gestational age at birth in singleton pregnancies : a multicentric prospective cohort study in the Brazilian population

Bill & Melinda Gates Foundation [OPP1107597], the Brazilian Ministry of Health, and the Brazilian National Council for Scientific and Technological Development (CNPq) [401615/20138]. The funders had no role in the design, development of the study, analysis, interpretation of data, writing the manuscript and in the decision to submit the article for publication. T.V.S. was supported by Coordenação de Aperfeiçoamento Pessoal de Nível Superior—CAPES (grant number 001).Peer reviewedPublisher PD

Phosphatidylserine Exposure by Toxoplasma gondii Is Fundamental to Balance the Immune Response Granting Survival of the Parasite and of the Host

Phosphatidylserine (PS) exposure on the cell surface indicates apoptosis, but has also been related to evasion mechanisms of parasites, a concept known as apoptotic mimicry. Toxoplasma gondii mimics apoptotic cells by exposing PS, inducing secretion of TGF-beta1 by infected activated macrophages leading to degradation of inducible nitric oxide (NO) synthase, NO production inhibition and consequently persisting in these cells. Here PS+ and PS− subpopulation of tachyzoites were separated and the entrance mechanism, growth and NO inhibition in murine macrophages, and mice survival and pathology were analyzed. Infection index in resident macrophages was similar for both PS subpopulations but lower when compared to the total T. gondii population. Growth in resident macrophages was higher for the total T. gondii population, intermediate for the PS+ and lower for the PS− subpopulation. Production of NO by activated macrophages was inhibited after infection with the PS+ subpopulation and the total populations of tachyzoites. However, the PS− subpopulation was not able to inhibit NO production. PS+ subpopulation invaded macrophages by active penetration as indicated by tight-fitting vacuoles, but the PS− subpopulation entered macrophages by phagocytosis as suggested by loose-fitting vacuoles containing these tachyzoites. The entrance mechanism of both subpopulations was confirmed in a non-professional phagocytic cell line where only the PS+ tachyzoites were found inside these cells in tight-fitting vacuoles. Both subpopulations of T. gondii killed mice faster than the total population. Clear signs of inflammation and no tachyzoites were seen in the peritoneal cavity of mice infected with the PS− subpopulation. Moreover, mice infected with the PS+ subpopulation had no sign of inflammation and the parasite burden was intense. These results show that PS+ and PS− subpopulations of T. gondii are necessary for a successful toxoplasma infection indicating that both subpopulations are required to maintain the balance between inflammation and parasite growth