Endocrine-disrupting chemicals (EDCs) are known to cause endocrine disruption (ED), developmental neurotoxicity (DNT), infertility and impaired embryo development. EDCs do therefore impose a threat to humans, wildlife and the environment. The present study investigated the effects of the reference compounds dihydrotestosterone, estradiol, benzo(a)pyrene, rosiglitazone, as well as the EDCs bisphenol F and perfluorooctanesulfonic acid at various concentrations on zebrafish embryos. The scientific questions of the study were to investigate how these environmental pollutants impact the development of zebrafish, what their molecular mechanisms are and what the link between ED and DNT is. Zebrafish embryos were exposed for 5 days to the compounds and various parameters on development were collected at different time points. The expression of 41 genes (qPCR) related to ED and DNT, and the levels of 23 steroids (LC-MS/MS) were determined. Gene correlations were determined with Pearson’s correlation test and paired t-tests were used to determine significantly altered gene activities. The significant gene expression changes were further related to the pathways of steroids in order to connect how gene activity impacted steroid levels. Exposure to estradiol, dihydrotestosterone and bisphenol F induced cyp19a1b expression which can affect personality traits. Perfluorooctanesulfonic acid interferes with thyroid hormone transport by binding to TTR causing profound effects on neurodevelopmental processes and cognitive functions. The compounds influenced genes that can disrupt endocrine systems which can cause neurodevelopmental impairments.

Endocrine disrupting chemicals (EDCs) are ubiquitous and their adverse impact on nature, wildlife and humans is extensively researched. We are constantly exposed to EDCs, such as the widespread and extensively researched Bisphenol A, as well as its substitute Bisphenol F, which is coming into wider use, even though it is much less is researched and limited information is available about its endocrine effects. Realistically, we are exposed to mixtures rather than single substances. In the Swedish Environmental Longitudinal, Mother and Child, Asthma and allergy (SELMA) study, the co-exposure of EDCs was assessed. Based on the SELMA cohort data, a study identified a mixture of EDCs, Mixture N1, which is associated with delayed language development. In recent years, it has been hypothesized that epigenetic alterations are one of the underlying mechanisms for the effect of EDC exposures. For example, EDC induced changes in DNA Methylation of the promoter region of a gene might lead to altered gene expression, which can result in adverse health effects. Several studies already indicate an impact of the formerly introduced chemicals/mixtures on the DNA methylation on genes such as NR3C1 and GRIN2B in animals. However, limited research is available on the impact on NR3C1 and GRIN2B in the early human brain, which is of interest since both genes are crucial for the development of the brain and altered gene expression often leads to adverse effects. This study aimed to investigate the impact of BPA, BPF and Mixture N1 on NR3C1 and GRIN2B in the developing human brain as well as establish a protocol for differentiation of human stem cells into neuroprogenitor cells that express GRIN2B and NR3C1. In the end stem cells were differentiated in vitro into neural progenitor cells (NPCs) using the protocol of Hosseini et al. (2020). During the differentiation, the cells were exposed to different concentrations of the former mentioned chemicals. Afterwards, RNA and DNA were extracted, followed by a qPCR and bisulfite-pyrosequencing to investigate the changes in gene expression and DNA methylation of NR3C1 and GRIN2B. This study established the differentiation protocol but revealed no significant results regarding the chemical exposure. However, some chemical exposures showed a clear tendency towards an impact of the chemicals on the gene expression and the DNA methylation. Furthermore, a negative correlation between DNA methylation at 2 CpG sites and gene expression in NR3C1 could be observed. In conclusion, the DNA methylation at promoter region in NR3C1 is important for the gene expression.

Additive manufacturing with the use of metals have been a steadily increasing field, being able to create products with a higher degree of complexity than traditional processing techniques. SLM is a popular AM process that uses metal powder as feedstock, and one of the key components of this process is the powder rheology. In recent years the use of a powder rheometer has been shown to be a good way of evaluating powder rheology of metal powders used within AM processes, but there is a clear lack of standardised tests and methods. In this study the Compressibility, Permeability, and Aeration test for 316L powders used within SLM processes was evaluated with a FT4 powder rheometer. 15 powders that had undergone printing in SLM processes were studied. This showed that the compressibility test had the best results in differentiating the bad preforming powders, thereafter the Aeration test. The Permeability test wasn’t able to differentiate the bad preforming powders with the settings used. This study demonstrates that some tests with a powder rheometer can evaluate the powder performance in SLM processes, but further research to evaluate the tests and standardise the settings are needed for clearer test results. / Additiv tillverkning med metall är ett område som stadigt ökat i intresse, främst på grund av möjligheten att producera produkter med en mycket högre grad av komplexitet i jämförelse med traditionella processmetoder. SLM är en populär AM process som använder metallpulver som råmaterial, och en av huvudkomponenterna för processen är pulvrets reologi. Under senare år har användningen av en pulver-reometer visat sig ett bra sätt att utvärdera pulver-reologi för metallpulver som används inom AM, men det finns en klar avsaknad av standardiserade test och metoder. I denna studie utvärderas Kompressabilitet, Permeabilitet, och Aerabilitet testen för 316L pulver producerade för SLM processer med en FT4 pulver-reometer. 15 pulver som genomgått SLM printing studerades. Studien visar att kompressabilitets testets utfall bäst överensstämde med det som setts under SLM processen, och bäst urskilde pulvren som fungerat dåligt att printa med, därefter Aerations testet. Permeabilitets testet kunde inte urskilja de sämre pulvren med de inställningarna som användes. Studien demonstrerar att vissa test och index samlade med ett pulver reometer är mer tillförlitliga än andra när det gäller för att utvärdera pulvrets prestanda inom SLM processer, men vidare forskning och studier krävs för att utvärdera testen och standardisera inställningar baserat på pulvret som testas.

A produção de kits para diagnóstico in vitro deve ser feita seguindo-se a legislação vigente de Boas Práticas de Fabricação e Controle (BPF). O objetivo deste trabalho foi elaborar um procedimento para desenvolvimento, produção e validação de um produto para diagnóstico in vitro, de acordo com a legislação vigente. Adotamos como modelo um kit imunoenzimático para Doença de Chagas. Dentro dos requisitos de BPF, a validação é uma etapa importante, pois tem por objetivos, dentre outros: auxiliar no estabelecimento de procedimentos de produção e controle de qualidade, avaliar desvios e dimensionar possíveis erros, avaliar o desempenho quanto à utilidade médica dos resultados obtidos e estabelecer condições ideais de uso. No estabelecimento dos requisitos para validação devem-se considerar as características do método utilizado, a utilidade clínica e diagnóstica dos resultados e as condições de uso do kit. Os parâmetros para validação devem ser definidos considerando a finalidade do uso do produto. Os resultados obtidos em três lotes pilotos demonstraram que o kit pode ser utilizado tanto com soro como com plasma, as amostras podem ser congeladas e descongeladas antes do uso por até 5 ciclos, o índice de concordância com kit comercial é de 0,9 (ótimo) e o kit mantém-se estável por pelo menos 7 dias à 37ºC, o que neste trabalho foi equivalente a pelo menos um ano na sua condição ideal de armazenamento de 2 a 8ºC. Além disso, o kit apresentou 100% de sensibilidade, 99% de especificidade, com coeficiente de variação 15,2% tanto na repetitividade como na reprodutibilidade de amostras positivas. Quanto à análise de interferentes, amostras hemolisadas e a presença de fator reumatóide podem interferir nos resultados e anticorpos anti-Leishmania na amostra podem dar reação cruzada. Conclui-se que o procedimento elaborado e o kit desenvolvido e validado atenderam aos requisitos pré-estabelecidos, de acordo com as regras de BPF vigentes. / The production of an in vitro diagnostic kit should be done following current Good Manufacturing Practices (GMP). The objective of this work was to establish a procedure for the development, production and validation of an in vitro diagnostic product in accordance with current regulations governing Medical Devices. An enzyme-linked immunoassay kit for Chagas\' disease was used as a model. Validation is a very important step contained within GMP requirements. Validation provides documented evidence that processes and product batches are consistent, it aids in the establishment of production and quality control procedures, evaluate deviations and identify possible mistakes, evaluate the performance and medical usefulness of the product based on the obtained results, and establish ideal conditions of use and storage. In order to establish validation requirements for product development, it is necessary to consider the characteristics of the assay method, the clinical and diagnostic usefulness of the results and the conditions of use of the kit. The parameters for validation should be defined considering the purpose of the use of the product. In the case of this Chagas assay, results obtained in three pilot lots demonstrated that the kit could be used with both serum and plasma, samples could be frozen and thawed before use for up to 5 cycles. The agreement index when compared with a commercially licensed kit is 0,9 (optimum correlation). The kit remained stable for at least 7 days at 37ºC, which is equivalent to at least one year stability in its ideal storage condition of 2 to 8ºC. The kit presented 100% sensitivity and 99% specificity, with variation coefficient of 15,2% for both repeatability and reproducibility of the positive samples. Interference analysis indicated that: hemolyzed samples and the presence of reumathoid factor could interfere with test results. Antibodies anti-Leishmania in the test sample can cross react with T. cruzi proteins. In conclusion: the established procedure for development and validation of chagas kit, and the actual developed and validated kit are in accordance with pre-established current GMP requirements.

Made available in DSpace on 2016-06-02T19:51:45Z (GMT). No. of bitstreams: 1 3188.pdf: 2904896 bytes, checksum: 9d486740c6a99b495aeb0b7d3f2aad98 (MD5) Previous issue date: 2010-08-23 / The raising of alligator in captivity began in Brazil as an option to protect the species and the land properties against invasions of alligator hunters. This activity was legalized by the decree 126/1990 of Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis (IBAMA). Increasingly, actions which are linked to the breeding of this animal in captivity are accomplished in the state of Mato Grosso by institutions like Sistema Brasileiro de Apoio às Micro e Pequenas Empresas (SEBRAE), Fundação Estadual do Meio Ambiente (FEMA) e IBAMA. These actions have as an aim to develop the productive chain of Jacaré do Pantanal, that have as the main product the skin of the animal and as sub-products, the meat, craft and animal food. The sub-product meat received detachment in this productive chain in 2008, due to the processing unit which belongs to the Cooperativa de Criadores de Jacaré do Pantanal (COOCRIJAPAN) that obtained the registration of the Serviço de Inspeção Federal (SIF), allowing the trading of alligator meat in national territory and abroad. To meet the quality requests and the security of the food, in general and at this unit, actions and procedures are necessary of the Quality Management. The objective of this study is to present a set of propositions for the Quality Management for the Processing Unit of COOCRIJAPAN, that meets the requests of the Good Manufacturing Practices (GMP), of NBR ISO 22000:2006 and of Hazard Analysis Critical Control Point (HACCP), which are considered specific methods for the Quality Management of Food Industries. With this objective, data were collected at COOCRIJAPAN, through interviews with the members of the unit and observations were made at the local to know the practices of the quality management of the Cooperative during the month of July of 2009. From the bibliographic revision and the field study, proposals was made for the Quality Management and Food Security for the Cooperative as well as a definition of a implementation plan for it. / A criação de jacaré em cativeiro teve início no Brasil como opção para proteção da espécie e das propriedades de terra contra invasões de caçadores de jacaré. Esta atividade recebeu suporte legal através da portaria 126/1990 do Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis (IBAMA). De maneira crescente, ações ligadas à criação deste animal em cativeiro são realizadas no Estado de Mato Grosso por órgãos como o Sistema Brasileiro de Apoio às Micro e Pequenas Empresas (SEBRAE), Fundação Estadual do Meio Ambiente (FEMA) e IBAMA. Essas ações objetivam desenvolver a cadeia produtiva do Jacaré do Pantanal, que tem como principal produto o couro do animal e como subprodutos a carne, artesanatos e ração animal. O subproduto carne recebeu destaque nesta cadeia produtiva em 2008, devido à unidade processadora, pertencente à Cooperativa de Criadores de Jacaré do Pantanal (COOCRIJAPAN), ter obtido o registro no Serviço de Inspeção Federal (SIF), possibilitando a comercialização da carne de jacaré em território nacional e no mercado externo. Para atender os requisitos da qualidade e da segurança do alimento, de modo geral e nesta unidade processadora, são necessárias ações e procedimentos da Gestão da Qualidade. O objetivo deste trabalho é apresentar propostas para a Gestão da Qualidade para a unidade processadora da COOCRIJAPAN, que atenda aos requisitos das Boas Práticas de Fabricação (BPF), da NBR ISO 22000:2006 e do Sistema de Análise de Perigos e Pontos Críticos de Controle (APPCC), que são consideradas abordagens específicas para a Gestão da Qualidade em indústrias alimentícias. Para isso, foram coletadas informações na COOCRIJAPAN, através de entrevistas com membros da unidade e feitas observações in loco para conhecer as práticas de gestão da qualidade da Cooperativa, durante o mês de julho de 2009. A partir da revisão bibliográfica e da pesquisa de campo, elaborou-se a proposta para a Gestão da Qualidade e da Segurança do Alimento para a Cooperativa, bem como um plano para implantação.

A produção de kits para diagnóstico in vitro deve ser feita seguindo-se a legislação vigente de Boas Práticas de Fabricação e Controle (BPF). O objetivo deste trabalho foi elaborar um procedimento para desenvolvimento, produção e validação de um produto para diagnóstico in vitro, de acordo com a legislação vigente. Adotamos como modelo um kit imunoenzimático para Doença de Chagas. Dentro dos requisitos de BPF, a validação é uma etapa importante, pois tem por objetivos, dentre outros: auxiliar no estabelecimento de procedimentos de produção e controle de qualidade, avaliar desvios e dimensionar possíveis erros, avaliar o desempenho quanto à utilidade médica dos resultados obtidos e estabelecer condições ideais de uso. No estabelecimento dos requisitos para validação devem-se considerar as características do método utilizado, a utilidade clínica e diagnóstica dos resultados e as condições de uso do kit. Os parâmetros para validação devem ser definidos considerando a finalidade do uso do produto. Os resultados obtidos em três lotes pilotos demonstraram que o kit pode ser utilizado tanto com soro como com plasma, as amostras podem ser congeladas e descongeladas antes do uso por até 5 ciclos, o índice de concordância com kit comercial é de 0,9 (ótimo) e o kit mantém-se estável por pelo menos 7 dias à 37ºC, o que neste trabalho foi equivalente a pelo menos um ano na sua condição ideal de armazenamento de 2 a 8ºC. Além disso, o kit apresentou 100% de sensibilidade, 99% de especificidade, com coeficiente de variação 15,2% tanto na repetitividade como na reprodutibilidade de amostras positivas. Quanto à análise de interferentes, amostras hemolisadas e a presença de fator reumatóide podem interferir nos resultados e anticorpos anti-Leishmania na amostra podem dar reação cruzada. Conclui-se que o procedimento elaborado e o kit desenvolvido e validado atenderam aos requisitos pré-estabelecidos, de acordo com as regras de BPF vigentes. / The production of an in vitro diagnostic kit should be done following current Good Manufacturing Practices (GMP). The objective of this work was to establish a procedure for the development, production and validation of an in vitro diagnostic product in accordance with current regulations governing Medical Devices. An enzyme-linked immunoassay kit for Chagas\' disease was used as a model. Validation is a very important step contained within GMP requirements. Validation provides documented evidence that processes and product batches are consistent, it aids in the establishment of production and quality control procedures, evaluate deviations and identify possible mistakes, evaluate the performance and medical usefulness of the product based on the obtained results, and establish ideal conditions of use and storage. In order to establish validation requirements for product development, it is necessary to consider the characteristics of the assay method, the clinical and diagnostic usefulness of the results and the conditions of use of the kit. The parameters for validation should be defined considering the purpose of the use of the product. In the case of this Chagas assay, results obtained in three pilot lots demonstrated that the kit could be used with both serum and plasma, samples could be frozen and thawed before use for up to 5 cycles. The agreement index when compared with a commercially licensed kit is 0,9 (optimum correlation). The kit remained stable for at least 7 days at 37ºC, which is equivalent to at least one year stability in its ideal storage condition of 2 to 8ºC. The kit presented 100% sensitivity and 99% specificity, with variation coefficient of 15,2% for both repeatability and reproducibility of the positive samples. Interference analysis indicated that: hemolyzed samples and the presence of reumathoid factor could interfere with test results. Antibodies anti-Leishmania in the test sample can cross react with T. cruzi proteins. In conclusion: the established procedure for development and validation of chagas kit, and the actual developed and validated kit are in accordance with pre-established current GMP requirements.