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Introdução: O novo coronavírus (SARS-CoV-2) é um patógeno que acomete principalmente o sistema respiratório, sendo o causador da COVID-19. Esse vírus pode afetar todos os grupos populacionais, entretanto indivíduos obesos são mais suscetíveis a evoluir com um quadro mais grave da doença. A obesidade pode ocasionar alterações do sistema imunológico, aumento do risco de infecções e aumento da taxa de mortalidade por COVID-19. Diante de maiores complicações relacionadas com a obesidade e a COVID-19, é necessário o tratamento específico para esses indivíduos no ambiente hospitalar. Portanto, o objetivo do estudo foi desenvolver um protocolo de atendimentonutricional a pacientes obesos com diagnóstico de COVID-19. Metodologia: A busca dos trabalhos foi realizada em base de dados PubMed, MEDLINE, LILACS, SciELO e Google Scholar, utilizando os descritores coronavirus, COVID-19, SARS-CoV-2, SARS-CoV como termo principal, associados a obesity, body composition, infflamation, adipose tissue, nutrition, critical illness. Resultados: foi estabelecido um protocolo para realização da avaliação e triagem de risco nutricional, com definição da via de alimentação pela equipe médica com o auxílio da equipe multiprofissional. As estimativas das necessidades calóricas e proteicas devem ser baseadas de acordo com a avaliação nutricional. Discussão: A avaliação, triagem e terapia nutricional adequadas são fundamentais para melhor recuperação de pacientes obesos. Devido a redução da ingestão alimentar e os sintomas gastrointestinais ocasionada pela COVID-19, é fundamental monitorar os sintomas e a tolerância a dieta ofertada. Conclusão: um protocolo de atendimento nutricional com padronizações de condutas nutricionais pode garantir menores complicações e contribuir para a redução da taxa de mortalidade. Introduction: The new coronavirus (SARS-CoV-2) is a pathogen that mainly affects the respiratory system, causing the disease COVID-19. This virus can affect all population groups, however obese individuals are more susceptible to evolving with a more severe disease. Obesity can cause changes in the immune system, increase the risk of infections and increase the mortality rate due to COVID-19. In the face of greater complications related to obesity and COVID-19, specific treatment for these individuals in the hospital environment is necessary. Therefore, the objective of the study was to develop a nutritional care protocol for obese patients diagnosed with COVID-19. Methodology: The search for the works was carried out in PubMed, MEDLINE, LILACS, SciELO and Google Scholar databases, using the keywords coronavirus, COVID-19, SARS-CoV-2, SARS-CoV as the main term, associated with obesity, body composition, infflamation, adipose tissue, nutrition, critical illness. Results: a protocol was established to carry out the assessment and screening of nutritional risk, with the definition of the feeding route by the medical team with the help of the multidisciplinary team. Estimates of calorie and protein requirements should be based on nutritional assessment. Discussion: Appropriate nutritional assessment, screening and therapy are essential for better recovery in obese patients. Due to the reduction in food intake and gastrointestinal symptoms caused by COVID-19, it is essential to monitor the symptoms and tolerance to the diet offered. Conclusion: a nutritional care protocol with standardization of nutritional behaviors can guarantee less complications and contribute to the reduction of the mortality rate. Introducción: El nuevo coronavirus (SARS-COV-2) es un patógeno que afecta principalmente al sistema respiratorio, siendo la causa de COVID-19. Este virus puede afectar a todos los grupos de población, sin embargo, los individuos obesos son más susceptibles a evolucionar con un marco más grave de la enfermedad. La obesidad puede causar cambios en el sistema inmunológico, mayor riesgo de infecciones y aumento de la tasa de mortalidad por Cuvid-19. Ante las más altas complicaciones relacionadas con la obesidad y la COVID-19, se necesita un tratamiento específico para estas personas en el entorno hospitalario. Por lo tanto, el objetivo del estudio fue desarrollar un protocolo de servicio nutricional a los pacientes obesos con un diagnóstico de COVID-19. Metodología: la búsqueda de trabajos se llevó a cabo en la base de datos PubMed, MEDLINE, LILACS, SCIELO y Google Scholar, utilizando los descriptores de Coronavirus, COVID-19, SARS-COV-2, SARS-COV como término principal, asociado con la obesidad, la composición corporal, Inflamación, tejido adiposo, nutrición, enfermedad crítica. Resultados: Se estableció un protocolo para la evaluación y clasificación de riesgo nutricional, con la definición de la ruta de alimentación por el equipo médico con la ayuda del equipo multiprofesional. Las estimaciones de las necesidades calóricas y proteicas deben basarse en la evaluación nutricional. Discusión: la evaluación nutricional adecuada, la clasificación y la terapia son fundamentales para una mejor recuperación de los pacientes obesos. Debido a la reducción de la ingesta de alimentos y los síntomas gastrointestinales causados ​​por COVID-19, es esencial monitorear los síntomas y la tolerancia a la dieta ofrecida. Conclusión: un protocolo de atención nutricional con estandarización de conducta nutricional puede garantizar complicaciones más pequeñas y contribuir a la reducción de la tasa de mortalidad. Introdução: O novo coronavírus (SARS-CoV-2) é um patógeno que acomete principalmente o sistema respiratório, sendo o causador da COVID-19. Esse vírus pode afetar todos os grupos populacionais, entretanto indivíduos obesos são mais suscetíveis a evoluir com um quadro mais grave da doença. A obesidade pode ocasionar alterações do sistema imunológico, aumento do risco de infecções e aumento da taxa de mortalidade por COVID-19. Diante de maiores complicações relacionadas com a obesidade e a COVID-19, é necessário o tratamento específico para esses indivíduos no ambiente hospitalar. Portanto, o objetivo do estudo foi desenvolver um protocolo de atendimentonutricional a pacientes obesos com diagnóstico de COVID-19. Metodologia: A busca dos trabalhos foi realizada em base de dados PubMed, MEDLINE, LILACS, SciELO e Google Scholar, utilizando os descritores coronavirus, COVID-19, SARS-CoV-2, SARS-CoV como termo principal, associados a obesity, body composition, infflamation, adipose tissue, nutrition, critical illness. Resultados: foi estabelecido um protocolo para realização da avaliação e triagem de risco nutricional, com definição da via de alimentação pela equipe médica com o auxílio da equipe multiprofissional. As estimativas das necessidades calóricas e proteicas devem ser baseadas de acordo com a avaliação nutricional. Discussão: A avaliação, triagem e terapia nutricional adequadas são fundamentais para melhor recuperação de pacientes obesos. Devido a redução da ingestão alimentar e os sintomas gastrointestinais ocasionada pela COVID-19, é fundamental monitorar os sintomas e a tolerância a dieta ofertada. Conclusão: um protocolo de atendimento nutricional com padronizações de condutas nutricionais pode garantir menores complicações e contribuir para a redução da taxa de mortalidade. Introduzione: il nuovo Coronavirus (SARS-COV-2) è un agente patogeno che colpisce principalmente il sistema respiratorio, essendo la causa del Covid-19. Questo virus può influenzare tutti i gruppi di popolazione, tuttavia gli individui obesi sono più suscettibili a evoluzione con un quadro più grave della malattia. L'obesità può causare cambiamenti nel sistema immunitario, aumentato il rischio di infezioni e un aumento del tasso di mortalità da CUVID-19. Di fronte a complicazioni più elevate relative all'obesità e a Covid-19, è necessario un trattamento specifico per questi individui nell'ambiente ospedaliero. Pertanto, l'obiettivo dello studio era quello di sviluppare un protocollo di servizio nutrizionale ai pazienti obesi con una diagnosi di Covid-19. Metodologia: la ricerca di posti di lavoro è stata tenuta nel database PubMed, Medline, Lilacs, SCielo e Google Scholar, utilizzando i descrittori di Coronavirus, Covid-19, SARS-Cov-2, SARS-COV come termine principale, associato all'obesità, alla composizione corporea, INFFLAMAZIONE, tessuto adiposo, nutrizione, malattia critica. Risultati: è stato istituito un protocollo per la valutazione e l'ordinamento del rischio nutrizionale, con la definizione della rotta di alimentazione dal team medico con l'aiuto del team multiprofessionale. Le stime delle esigenze caloriche e proteiche dovrebbero essere basate sulla valutazione nutrizionale. Discussione: valutazione nutrizionale adeguata, smistamento e terapia sono fondamentali per un migliore recupero dei pazienti obesi. A causa della riduzione del consumo di cibo e dei sintomi gastrointestinali causati da Covid-19, è essenziale monitorare i sintomi e la tolleranza della dieta offerta. Conclusione: un protocollo di assistenza nutrizionale con la standardizzazione della condotta nutrizionale può garantire complicazioni più piccole e contribuire alla riduzione del tasso di mortalità.

The text deals with Management of Municipal Systems of Education, Secondary Education and training in the Pará Amazon in the context of the federative pact. Aims to discuss the category autonomy in the collaboration regime as the basis of the National Education System to the formulation of public policies. Through bibliographic and documentary research, we focused on the federative relationship to effectively manage public high school policies in the face of controversies and various problems that permeate the formation. We consider that the configuration of the municipal federated entity corroborates the urgency of the proper articulation in the SNE that needs to pursue a collective stance of resistance in the course of policies, expressed by regulations that subject the teacher to the fulfillment of requirements that support the capitalist system.

Acute myeloid leukemia (AML) is a complex hematological disorder characterized by blockage of differentiation and high proliferation rates of myeloid progenitors. Anthracycline and cytarabine‑based therapy has remained the standard treatment for AML over the last four decades. Although this treatment strategy has increased survival rates, patients often develop resistance to these drugs. Despite efforts to understand the mechanisms underlying cytarabine resistance, there have been few advances in the field. The present study developed an in vitro AML cell line model resistant to cytarabine (HL‑60R), and identified chromosomal aberrations by karyotype evaluation and potential molecular mechanisms underlying chemoresistance. Cytarabine decreased cell viability, as determined by MTT assay, and induced cell death and cell cycle arrest in the parental HL‑60 cell line, as revealed by Annexin V/propidium iodide (PI) staining and PI DNA incorporation, respectively, whereas no change was observed in the HL‑60R cell line. In addition, the HL‑60R cell line exhibited a higher tumorigenic capacity in vivo compared with the parental cell line. Notably, no reduction in tumor volume was detected in mice treated with cytarabine and inoculated with HL‑60R cells. In addition, western blotting revealed that the protein expression levels of Bcl‑2, X‑linked inhibitor of apoptosis protein (XIAP) and c‑Myc were upregulated in HL‑60R cells compared with those in HL‑60 cells, along with predominant nuclear localization of the p50 and p65 subunits of NF‑κB in HL‑60R cells. Furthermore, the antitumor effect of LQB‑118 pterocarpanquinone was investigated; this compound induced apoptosis, a reduction in cell viability and a decrease in XIAP expression in cytarabine‑resistant cells. Taken together, these data indicated that acquired cytarabine resistance in AML was a multifactorial process, involving chromosomal aberrations, and differential expression of apoptosis and cell proliferation signaling pathways. Furthermore, LQB‑118 could be a potential alternative therapeutic approach to treat cytarabine‑resistant leukemia cells.

A non-synonymous mtDNA mutation, m.3395A > G, which changes tyrosine in position 30 to cysteine in p.MT-ND1, was found in several patients with a wide range of clinical phenotypes such as deafness, diabetes and cerebellar syndrome but no Leber’s hereditary optic neuropathy. Although this mutation has already been described, its pathogenicity has not been demonstrated. Here, it was found isolated for the first time, allowing a study to investigate its pathogenicity. To do so, we constructed cybrid cell lines and carried out a functional study to assess the possible consequences of the mutation on mitochondrial bioenergetics. Results obtained demonstrated that this mutation causes an important dysfunction of the mitochondrial respiratory chain with a decrease in both activity and quantity of complex I due to a diminution of p.MT-ND1 quantity. However, no subcomplexes were found in cybrids carrying the mutation, indicating that the quality of the complex I assembly is not affected. Moreover, based on the crystal structure of p.MT-ND1 and the data found in the literature, we propose a hypothesis for the mechanism of the degradation of p.MT-ND1. Our study provides new insights into the pathophysiology of mitochondrial diseases and in particular of MT-ND1 mutations.

Ten-eleven-translocation 2 (TET2) belongs to the TET protein family that catalyzes the conversion of 5-methylcytosine into 5-hydroxymethylcytosine and plays a central role in normal and malignant adult hematopoiesis. Yet the role of TET2 in human hematopoietic development remains largely unknown. Here, we show that TET2 expression is low in human embryonic stem cell (ESC) lines and increases during hematopoietic differentiation. shRNA-mediated TET2 knockdown had no effect on the pluripotency of various ESCs. However, it skewed their differentiation into neuroectoderm at the expense of endoderm and mesoderm both in vitro and in vivo. These effects were rescued by reintroducing the targeted TET2 protein. Moreover, TET2-driven differentiation was dependent on NANOG transcriptional factor. Indeed, TET2 bound to NANOG promoter and in TET2-deficient cells the methylation of the NANOG promoter correlated with a decreased in NANOG expression. The altered differentiation resulting from TET2 knockdown in ESCs led to a decrease in both the number and the cloning capacities of hematopoietic progenitors. These defects were due to an increased apoptosis and an altered gene expression profile, including abnormal expression of neuronal genes. Intriguingly, when TET2 was knockdown in hematopoietic cells, it increased hematopoietic development. In conclusion, our work suggests that TET2 is involved in different stages of human embryonic development, including induction of the mesoderm and hematopoietic differentiation. Stem Cells 2014;32:2084–2097

A partir da tradução do poema hexamétrico Europa atribuído a Mosco de Siracusa, o presente artigo pretende discutir a ênfase dada nessa composição aos elementos eróticos. Ao detectar as alusões no poema a narrativas épicas anteriores, tais como a Odisseia, o Catálogo das Mulheres e o Hino Homérico a Deméter, serão feitos alguns comentários sobre a maneira como Mosco explora o erotismo já presente no epos arcaico, conferindo-lhe um enfoque central.

1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) intoxicated mice have been widely used to model the loss of dopaminergic neurons. As this treatment leads to basal ganglia degeneration, it was proposed that MPTP mice could be used as a model of Leigh syndrome. However, this mitochondrial pathology is biochemically characterized by a respiratory chain dysfunction. To determine if MPTP can affect in vivo mitochondria function, we measured the activities of mitochondrial respiratory chain complexes in several tissues. Our results show that MPTP affects mainly mitochondrial respiratory chain complex IV, as found in Leigh Syndrome, confirming that acute MPTP intoxicated mice are a good model of Leigh Syndrome.

Essa dissertação tem como objetivos a tradução ao português brasileiro e o estudo de duas declamações, quais sejam O jovem herói (Decl. 5) e O velho sovina (Decl. 6), creditados a Corício de Gaza, um professor de retórica cuja produção se situou na primeira metade do séc. VI d.C. Primeiramente, apresentam-se as traduções. Em seguida, o estudo, que é dividido em dois blocos: o primeiro capítulo busca contextualizar o corpus tendo em vista a conjuntura histórico-literária na qual ambas as declamações se inserem. O segundo capítulo corresponde à analise descritiva e interpretativa das principais características estilísticas, linguística e argumentativa de ambos os textos. The main goals of this research are the translation into Brazilian Portuguese and an introductory study of two declamations, The Young Hero (Decl. 5) and The Miser Old Man (Decl. 6), credited to Choricius, a teacher of rhetoric who worked and lived in the 6th century Gaza. Firstly, I present the translation, then the study, which is divided into two sections: in the first chapter I contextualize both declamations historically and in the literary sense. In the second chapter I present a descriptive and interpretative analysis of the main stylistic, linguistic and argumentative characteristics of both texts.

TET2 converts 5-methylcytosine to 5-hydroxymethylcytosine (5-hmC) in DNA and is frequently mutated in myeloid malignancies, including myeloproliferative neoplasms. Here we show that the level of 5-hmC is decreased in granulocyte DNA from myeloproliferative neoplasm patients with TET2 mutations compared with granulocyte DNA from healthy patients. Inhibition of TET2 by RNA interference decreases 5-hmC levels in both human leukemia cell lines and cord blood CD34+ cells. These results confirm the enzymatic function of TET2 in human hematopoietic cells. Knockdown of TET2 in cord blood CD34+ cells skews progenitor differentiation toward the granulomonocytic lineage at the expense of lymphoid and erythroid lineages. In addition, by monitoring in vitro granulomonocytic development we found a decreased granulocytic differentiation and an increase in monocytic cells. Our results indicate that TET2 disruption affects 5-hmC levels in human myeloid cells and participates in the pathogenesis of myeloid malignancies through the disturbance of myeloid differentiation.

DOI: 10.14684/intertech.13.2014.422-426 The study evaluates surface soil samples from a nearby region currently inhabited an area previously occupied by disposal of solid waste from various sources ( landfill ) in order to verify a possible efficiency in agricultural productivity and recovery of this land to the inhabitants of these avoiding probable disease areas . This research was carried out physico - chemical and chemical (pH, organic matter, exchangeable calcium and magnesium, exchangeable aluminum, exchangeable hydrogen and aluminum). The parameters studied were those recommended to the relevant literature , taking into account the results obtained in this work it is concluded that the studied soil in the neighborhood Beira Rio ( Tucurui - PA ) has low fertility , by presenting acidic character possessing high availability of aluminum, thus raising the toxicity of the soil . While the content of organic matter varies intensely at all points due to the continuous removal of the solid residues from this area did not allow the formation horizons . Index Terms - soil profiles, dump Riverside, fertility.

O estudo focou a dimensão do contexto da comunidade de São João do Médio Itacuruçá, Abaetetuba/PA, na construção identitária dos sujeitos ribeirinhos quilombolas. A realidade se entrelaça entre rios, mata, estradas, memória e história de luta. Objetiva, portanto, provocar discussões pertinentes ao conceito ribeirinho quilombola. Os sujeitos vivem, moram, lutam pela sobrevivência à margem dos rios e igarapés, no entanto, trazem marcas identitárias quilombola. Como se identificam? Ribeirinhos ou quilombolas? Ou ribeirinhos quilombolas? O trabalho foi de cunho qualitativo, com trabalho de campo com aporte em fontes bibliográficas pertinentes à temática, tendo por base livros, dissertações, teses, artigos, sites. Os resultados pontuam a relevância da discussão do conceito ribeirinho quilombola vivenciado no cotidiano desses sujeitos. The study focused on the dimension of the context of the community of St. John’s Midfield Itacuruçá, Abaetetuba / PA, in the relevance of the riparian quilombolas subjects’ identity construction. The reality intertwines between rivers, woods, roads, memory and stories of struggles. Objective therefore lead discussions relevant to the concept riparian quilombola. The subjects live, reside, struggling to survive on the margins of rivers and streams, however, bring identity marks quilombolas. As identify? Riparian or quilombolas? Or riparian quilombolas? The study was a qualitative one, with fieldwork with investments in bibliographical sources relevant to the topic based on books, dissertations, theses, articles, websites. The results punctuate the importance of discussing the concept riparian quilombola experienced in everyday life these subjects. UFPA - Universidade Federal do Pará

UFPA - Universidade Federal do Pará O estudo foca a relação entre os saberes ribeirinhos quilombolas e Educação de Jovens e Adultos investigando as possibilidades com que os saberes ribeirinhos quilombolas da comunidade de São João do Médio Itacuruçá se relaciona com a Educação de Jovens e Adultos e como se inserem no contexto escolar. De maneira específica buscamos identificar a dimensão do contexto da comunidade de São João destacando os saberes ribeirinhos quilombolas; investigar a relação dos saberes ribeirinhos quilombolas com a Educação de Jovens e Adultos e proporcionar reflexões sobre o desafio da Educação de Jovens e Adultos frente as perspectivas pedagógicas no contexto escolar desses sujeitos. O trabalho foi de cunho qualitativo, com trabalho de campo na comunidade de São João do Médio Itacuruçá com aporte em fontes bibliográficas pertinentes à temática tendo por base livros, dissertações, teses,artigos, sites. Os resultados pontuam a relevância dos saberes ribeirinhos quilombolas na comunidade e, como o quefazer da Educação de Jovens e Adultos,que se constitui meio à organização política vem contribuindo nos novos direcionamentos de uma educação voltada às especificidades de seus sujeitos. Entretanto a comunidade de São João com os saberes próprios diante de suas organizações políticas, ainda não conseguiu inovar pedagogicamente na Educação de Jovens e Adultos, embora perspectivas de mudanças já se façam presente. The study focuses on the relationship between riparian quilombola knowledge and the Youth and Adults Education investigating the possibility with which the riparian quilombola knowledge of the community of St. John of the Middle Itacuruçá relates to Youth and Adults Education and how they fit into the school context in that community. Specifically we seek to identify the size of the context of the community of St. John highlighting the knowledge of riparian quilombola; investigate the relationship of riparian quilombola knowledge with the Youth and Adults Education, and offer reflections on the challenge of Education for Young Adults and the front pedagogical perspectives in the context of school subjects. The study is qualitative, with field research in the community of St. John of the Middle Itacuruçá with investments in bibliographic sources relevant to the topic based on books, dissertations, theses, articles, websites. The results punctuate the importance of the riparian quilombola knowledge in the community and, how the duties of the Education of the Youth and Adults, which is in the midst of political organization, has contributed to new directions of an education geared to the specifics of their subjects. However, the community of St. John, with the knowledge of their own political organizations, still can not innovate pedagogically in Youth and Adults Education, although prospects for change are present.

Orientador: Fernando Ferreira Costa Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas Resumo: Policitemia vera (PV) é uma síndrome mieloproliferativa (SMP) crônica que ocorre por proliferação clonal de progenitores hematopoéticos multipotenciais. Pacientes com PV apresentam expansão das três principais linhagens mielóides na medula óssea, levando à produção aumentada de hemácias, granulócitos e plaquetas. Características importantes de PV incluem elevação de hematócrito em presença de níveis normais ou diminuídos de eritropoetina (EPO) e a formação de colônias eritróides endógenas (CEE). A mutação JAK2 V617F, presente na maioria dos pacientes com PV, causa ativação constitutiva de JAK2 e parece ser responsável pelo fenótipo observado. Apesar disso, as mudanças transcricionais desencadeadas pela mutação ainda não foram completamente caracterizadas. Neste trabalho, utilizou-se Serial Analysis of Gene Expression (SAGE) para realizar um estudo de expressão gênica global em células da medula óssea de um paciente com PV portador da mutação, ao diagnóstico e em células normais de doadores saudáveis. Genes com expressão aumentada em PV estão envolvidos em processos biológicos importantes, como transdução de sinal, diferenciação e proliferação celular, ciclo celular, apoptose, resposta imune e regulação da transcrição. JUNB foi um dos genes identificados e, usando reação em cadeia da polimerase quantitativa em tempo real (qRT-PCR), detectou-se expressão aumentada de JUNB em dois outros pacientes com PV JAK2 V617F-positivos. Por meio de linhagens murinas Ba/F3-REPO e culturas primárias de eritroblastos humanos, observou-se que JUNB é expresso após adição de EPO e que a proteína JunB é constitutivamente induzida por JAK2 V617F. Além disso, interferência na expressão de JUNB diminuiu o potencial clonogênico e proliferativo de progenitores eritróides humanos Ainda, em PV, a eritropoese causada por JAK2 V617F mostrou-se mais sensível à diminuição de expressão de JunB do que a eritropoese normal. Assim, esses resultados sugerem que JunB tenha um papel fundamental no desenvolvimento de SMPs causadas por JAK2 V617F Abstract: Polycythemia vera (PV) is a chronic myeloproliferative disorder that arises through clonal proliferation of multipotent hematopoietic progenitors. PV patients present bone marrow trilineage expansion, leading to increased production of mature red cells, granulocytes and platelets. Important PV features are elevated red cell mass, despite normal or subnormal erythropoietin (EPO) levels, and endogenous erythroid colony (EEC) formation. The JAK2 V617F mutation, present in the majority of polycythemia vera (PV) patients, causes a JAK2 constitutive activation and seems to be responsible for the PV phenotype. However, the transcriptional changes triggered by the mutation have not yet been totally characterized. Serial analysis of gene expression (SAGE) was used to perform a large scale gene expression study in bone marrow cells of a newly-diagnosed PV patient harboring the JAK2 V617F mutation and in normal bone marrow cells from healthy donors. Genes overexpressed in PV are involved in important biological processes, such as signal transduction, cellular differentiation, cellular proliferation, cell cycle, apoptosis, immune response and transcriptional regulation. JUNB was one of the genes up-regulated in PV and overexpression of JUNB was also detected by quantitative real time polymerase chain reaction (qRT-PCR) in hematopoietic cells of another two JAK2 V617F PV patients. Using Ba/F3-EPOR cell lines and primary human erythroblast cultures, JUNB was found to be expressed after Epo addition and JunB protein was shown to be constitutively induced by JAK2 V617F. In addition, JUNB knock down reduced the clonogenic and proliferative potential of human erythroid progenitors. Furthermore, in PV, the JAK2 V617F driven erythropoisis was more sensitive to JunB downregulation than normal erythropoiesis. Hence, these results demonstrate that JunB may play a major role in the development of JAK2 V617F myeloproliferative disorders Doutorado Medicina Experimental Doutor em Fisiopatologia Médica

Abstract 2418 TET2 belongs to the TET family proteins that catalyze 5-methylcytosine (5mc) to 5-hydroxymethylcytosine (hmc) and plays an important role in normal and malignant adult hematopoiesis. The role of TET2 in human hematopoietic development remains unknown. Here we show that TET2 is expressed at low level in human embryonic stem (hES) cell lines and that its expression increases during hematopoietic differentiation in three different hES. TET2 knockdown does not modify hmc level and pluripotent properties of ES cells. However TET2 depletion by two different shRNA skewed differentiation into neuroectoderm at the expense of endoderm and mesoderm. This was associated with a decrease or an increase in promoter methylation of neuroectoderm and meso/endoderm genes, respectively during hES specification. Subsequently, we observed a decrease in hematopoietic progenitors (CD34+CD43+) and their cloning capacities due to a marked increase in apoptosis. Alteration of hematopoietic differentiation was coupled with a profound alteration in gene expression with up and down regulated genes including the abnormal expression of neuronal genes in hematopoietic cells. Thus our results suggest that TET2 regulates embryonic development by inhibiting neuroectoderm specification and enabling hematopoietic differentiation in hES cells. Disclosures: No relevant conflicts of interest to declare.

Abstract 1584 TET2, a gene frequently mutated in myeloid malignancies, encodes an oxygenase that may convert 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) in hematopoietic cells. Using high performance liquid chromatography coupled to tandem mass sprectrometry (HPLC-MS-MS), we found lower levels of 5hmC in erythroblasts from patients with myeloproliferative neoplasms and TET2 mutations than in erythroblasts from non mutated patients. To study the function of TET2 during myelopoiesis, we used lentiviruses expressing short hairpin RNA (shRNA) designed to inhibit TET2 expression in CD34+ cells from normal bone marrow or umbilical cord blood. TET2 mRNA was knocked down by 56 to 74% in several human leukemia cell lines expressing shRNA-TET2, compared to cell lines expressing scramble shRNA. TET2 expression was then assessed in these cell lines by western blot, which showed a 49% reduction of TET2 protein in TF1 cell line, and 80 to 90% reduction in Kasumi, Uke1, and Mo7E cell lines. In addition, knock down of TET2 led to a 73% decrease in 5hmC in Mo7E cell line DNA whereas 5mC and cytosine contents remained unchanged as measured by HPLC-MS-MS. These results indicate that TET2 has a role in the hydroxylation of 5mC in human cells of hematopoietic origin. We then studied the consequences of TET2 knock-down in umbilical cord blood CD34+ cells in vitro. We observed a skewing of CD34+CD38- progenitor differentiation toward the myeloid lineage (52 +/− 4% of the TET2 knock down cells versus 35 +/− 3% of the control cells, n=3, p=0.001) at the expense of lympho-myeloid development and B cell and natural killer (NK) lymphoid lineages in a B/myeloid/NK culture condition. Methylcellulose in the presence of EPO, IL3, SCF, G-CSF showed greater numbers of CFU-G/GM (66 +/− 4 versus 58 +/− 3 colonies per 1000 input CD34+ cells) and lower numbers of BFU-E (60 +/−19 versus 78 +/− 19) in TET2 knock-down samples than in control samples (n= 4, p Disclosures: No relevant conflicts of interest to declare.

Submitted by Boris Flegr (boris@uerj.br) on 2021-01-05T19:37:36Z No. of bitstreams: 1 TESE_FINAL_PUBLICADA_Renata_Lyrio_Rafael_Baptista.pdf: 1736690 bytes, checksum: da6823e80401b6785071617f6aab06f2 (MD5) Made available in DSpace on 2021-01-05T19:37:36Z (GMT). No. of bitstreams: 1 TESE_FINAL_PUBLICADA_Renata_Lyrio_Rafael_Baptista.pdf: 1736690 bytes, checksum: da6823e80401b6785071617f6aab06f2 (MD5) Previous issue date: 2018-03-12 Introduction: Although most cases of AML are sporadic, a small subgroup is associated with the presence of germ mutations. The 2016 review of the WHO Classification of 2008 included these cases in a session of myeloid neoplasia with presence of germ mutations. These patients should have a different management since the indication of related allogeneic HSCT makes it mandatory to exclude mutation in the donor sibling. In addition, family members should receive genetic counseling. Objectives: a) To establish the mutational status of the CEBPA, RUNX1, GATA2, TERT, TERC genes in patients with AML undergoing allo-HSCT in CEMO-INCA. During this work, a pair of twins discordant for the diagnosis of AML was identified. The rarity of this fact and the potential use of the model to try to understand the pathogenesis of AML lead to another goal to be developed; b) To evaluate the exome of identical twin siblings discordant for the diagnosis of AML identified in the studied cohort. Methods: The detection of mutations in the genes RUNX1, CEBPA, GATA2, TERT and TERC at 58 patients with the diagnosis of AML was performed with the implementation of the MLPA (Multiplex Ligation-dependent Probe Amplification). The evaluation of exomes was performed through bioinformatic analysis. Results: The evaluation of the mutational status of the CEBPA, RUNX1, GATA2, TERT, TERC genes did not identify any mutation. Comparison of the exomes of the identical twin discordant to AML identified 9 genes mutated in the patient and 2 genes mutated in the healthy sibling with possible clinical relevance. Genes HJURP, ABCE1, PCDHA11, AJM1, TAS2R19, AMER2, FNDC3A, SIN3A and L1CAM are mutated in the patient. RBM5 and ATXN3 are mutated in the health twin. Conclusion: At the exome analysis of the discordant twins pair in AML the mutations found in here could be the result of genetic instability lead by malignance. More experiments are needed to confirm the role of each mutation found in discordance between the pair of twins in this initial genetic screening. Introdução: Apesar da maioria dos casos de LMA serem esporádicos, um pequeno subgrupo está associado à presença de mutações germinativas. Na revisão de 2016 da Classificação da OMS de 2008 foram incluídas as neoplasias mieloides com presença de mutações germinativas. Esses pacientes devem ser submetidos a uma abordagem terapêutica diferenciada, uma vez que a indicação de TCTH alogênico aparentado torna obrigatória a exclusão da mutação no irmão doador. Além disso, os familiares devem receber aconselhamento genético. Objetivos: a) Estabelecer o status mutacional dos genes CEBPA, RUNX1, GATA2, TERT, TERC em pacientes com LMA submetidos à TCTH alogênico no CEMO-INCA. No decorrer desse estudo foi identificado um par de gêmeos discordantes para o diagnóstico de LMA. A raridade desse fato e o potencial uso do modelo para tentar entender a patogênese da LMA fez com que um outro objetivo fosse desenvolvido; b) Avaliar os exomas de irmãos gêmeos idênticos discordantes para o diagnóstico de LMA identificados na coorte estudada. Métodos: A detecção de mutações nos genes CEBPA, RUNX1, GATA2, TERT e TERC em 58 pacientes com diagnóstico de LMA foi realizada pela técnica de MPLA (Amplificação de primer por ligação dependente multiplex). A avaliação dos exomas foi realizada por meio de análise bioinformática. Resultados: Na avaliação do status mutacional dos genes CEBPA, RUNX1, GATA2, TERT, TERC através da técnica de MPLA não foi identificada nenhuma mutação. A comparação do exoma dos gêmeos idênticos discordantes para LMA identificou 116 variantes, dessas foram selecionadas mutações em nove genes no paciente e em dois genes no irmão saudável. A categoria funcional desses genes (HJURP, ABCE1, PCDHA11, AJM1, TAS2R19, AMER2, FNDC3A, SIN3A e L1CAM) e sua possível relevância no processo de transformação é discutida. RBM5 e ATXN3 sã mutados no gêmeo saudável. Conclusão: Na análise de exoma em par de gêmeos discordantes para LMA, as mutações encontradas podem ser o resultado da instabilidade genética gerada por malignidades. Mais estudos são necessários para conformar o papel de cada mutação encontrada na discordância entre o par de gêmeos neste rastreamento genético inicial.