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A lesão por pressão (LPP) e a dermatite relacionada à incontinência (DAI) são lesões que afetam a pele, principalmente, em pacientes hospitalizados, acamados e em situações críticas. Esses eventos costumam ter manifestações clínicas muito semelhantes, sendo imprescindível que os profissionais de saúde saibam diferenciá-los para prestar o atendimento adequado a cada situação. Este estudo visa avaliar o conhecimento da equipe de Enfermagem sobre a LPP nos estágios 1 e 2 e DAI em relação à definição, avaliação, identificação, prevenção e ao tratamento. Trata-se de um estudo descritivo, exploratório, com análise quantitativa dos dados, apresentados em tabelas. A pesquisa foi realizada com a equipe de enfermagem da Santa Casa de Misericórdia de Paranaíba- MS que é uma instituição filantrópica sem fins lucrativos. Os resultados desta pesquisa permitiram identificar a fragilidade dos conhecimentos dos enfermeiros, técnicos de enfermagem e auxiliares de enfermagem relacionados à temática que norteou o estudo. Frente às informações apresentadas, este estudo corrobora o déficit de conhecimento da equipe de enfermagem acerca do tema em relação à prevenção. KNOWLEDGE ABOUT DERMATITIS ASSOCIATED WITH INCONTINENCE AND PRESSURE INJURY OF THE NURSING TEAM ABSTRACT Pressure ulcer (PU) and incontinence-associated dermatitis (IAD) are lesions that irritate the skin, mainly in hospitalized, bedridden patients who are in critical conditions. These events usually present remarkably similar clinical manifestations, it is crucial that healthcare practitioners know how to distinguish them to provide better assistance to each situation. This paper aims at evaluating the knowledge of the Nursing team of PU stages 1 and 2 and IAD regarding definition, evaluation, identification, prevention, and treatment. It is a descriptive, exploratory study with quantitative data analysis, shown in tables. The research was carried out with the nursing team from Santa Casa de Misericórdia de Paranaína-MS which is a philanthropic non-profitable organization. The results from this research enabled the identification of the nursing personnel’s poor knowledge regarding the subject that guided this study. In the light of the information provided, this study supports the nursing team’s poor knowledge of the issue regarding prevention. Keywords: Pression Ulcer. Dermatitis. Nursing Team. Incontinence.

O objetivo deste estudo foi caracterizar o perfil do consumidor de produtos lácteos de origem bubalina e bovina nos municípios de Soure e Salvaterra (Ilha do Marajó-PA), quanto ao perfil sociodemográfico, ao nível de conhecimento sobre qualidade higiênico-sanitária e sobre as fraudes nesses produtos. Para coleta dos dados foram aplicados questionários em voluntários, todos escolhidos ao acaso. Os dados obtidos foram analisados de forma descritiva e posteriormente submetidos à análise de correspondência múltipla (ACM). A coleta de dados resultou em um total de 403 entrevistas. Nossos resultados demonstraram que para consumo de produtos de origem bubalina os entrevistados eram em sua maioria do sexo masculino, acima de 40 anos, que consomem principalmente dos produtores rurais e os que consomem produtos bovinos eram mulheres com idades entre 21 a 30 anos, que consomem em estabelecimentos comerciais como supermercados e padarias. As principais fontes de proteína animal derivada do leite na região do arquipélago do Marajó são principalmente produzidas a partir da bubalinocultura.

Introdução: De acordo com a Organização Mundial da Saúde o Diabetes Mellitus Gestacional é disfunção metabólica pelo qual há uma intolerância a carboidratos de gravidade variável. Diversos estudos têm apontado que essa disfunção é muito comum no período gestacional, atingindo cerca de 3 a 25% das gestantes. Objetivo: Analisar os principais aspectos envolvendo a mellitus gestacional e identificar o papel do profissional de farmácia nesse caso. Metodologia: Este trabalho trata-se de um estudo descritivo e exploratório embasado em revisão de literatura, realizando pesquisa e recolhimento de dados científicos organizados por meio de apuração de artigos associados com o objeto do estudo. Resultados Esperados: O DMG apresenta riscos para a mãe, feto e o recém-nascido, geralmente, é diagnosticado no 2º ou 3º trimestres. Pode ser transitória ou persistir após o parto, caracterizando-se como importante fator de risco para o desenvolvimento de Diabetes Mellitus tipo 2. A prevalência varia de 1% a 14%, dependendo da população estudada e dos critérios diagnósticos utilizados. Considerações finais: A metformina que é um remédio hipoglicemiante oral indicado principalmente para o tratamento da diabetes mellitus gestacional, age reduzindo os níveis de açúcar no sangue, para níveis mais próximos do normal. O cuidado farmacêutico promove a recuperação da saúde integralizada com o uso racional de medicamentos a fim de melhorar a qualidade de vida do paciente, em relação a medidas não farmacológicas, o farmacêutico durante a Atenção Primária (AT) à gestante pode orientar sobre ações em saúde que visem melhorar a qualidade de vida da paciente.

Purpose: to learn the deaf people’s perspective on the improvements needed in this population’s health care. Methods: an observational, cross-sectional study conducted with 124 deaf people who answered a semi-structured questionnaire to characterize the sample (age, sex, and means of communication) and collect answers to the open question: “Do you have any suggestion to improve the health care for the deaf?” - which was answered either in writing or in the Brazilian Sign Language (Libras). A descriptive analysis was conducted to characterize the sample, as well as a quantitative and qualitative content analysis (thematic-categorical), to identify categories and occurrence frequency of the content in the answers to the open question. Results: the sample’s mean age was 44 years (standard deviation 15, minimum 18, and maximum 70 years), 65% were women, and 78% used Libras to communicate. Most of the participants (83%) answered the open question in writing. Six theme categories were identified: 1) Needed improvements; 2) Communication barriers); 3) Health promotion; 4) Autonomy; 5) Achievements; and 6) Law. Conclusion: the results reveal a need for improvements in the health care of deaf people. The perspective of the studied deaf population is based on their desire to gain autonomy, overcome communication barriers, and have access to information, aiming at health promotion. RESUMO Objetivo: conhecer a perspectiva do surdo quanto às melhorias necessárias no atendimento à saúde para essa população. Métodos: estudo observacional transversal realizado com 124 surdos que responderam a questionário semiestruturado, visando caracterizar amostra (idade, sexo e tipo de comunicação) e coletar resposta à questão aberta: “Você tem alguma sugestão para melhorar o atendimento de saúde ao surdo?, respondida nas modalidades escrita ou Língua Brasileira de Sinais (Libras). Realizou-se análise descritiva para caracterização da amostra e análise quanti-qualitativa de conteúdo (temático-categorial), para identificação de categorias e frequência de ocorrência dos conteúdos das respostas à questão aberta. Resultados: amostra apresentou média de idade de 44 anos (desvio padrão: 15, mínima de 18 e máxima de 70 anos), sendo 65% mulheres e 78% usuários de Libras. A maioria dos participantes (83%) respondeu à questão aberta na modalidade escrita. Foram identificadas seis categorias temáticas: 1) Necessidade de melhorias; 2) Barreiras de comunicação); 3) Promoção da saúde; 4) Autonomia; 5) Conquistas; e 6) Legislação. Conclusão: os resultados evidenciaram necessidade de melhorias no atendimento à saúde do usuário surdo. A perspectiva da população surda estudada é pautada no desejo de autonomia, de superação das barreiras de comunicação e de acesso à informação visando promoção da saúde.

Purpose: to investigate the satisfaction of deaf people in relation to the health services, to characterize the sample regarding sociodemographic, socioeconomic, and self-perception of deafness, and to verify if there is an association between satisfaction with care, communication, professionals, and self-perception of deafness. Methods: an observational, cross-sectional study conducted with a semi-structured questionnaire in 74 deaf adults. A descriptive analysis of the characterization of the sample and attendance, and an analysis of the association between satisfaction, professionals, self-perception, service used, and schooling level was performed, using the chi-square test. The level of significance adopted was 5%. Results: the sample was composed mainly of women (66.2%), 18 to 28 years old (46.5%), of socioeconomic class D (51.3%), having finished high school (56.76%), and employed as an office assistant or typist (90%), 63.5% self-declared as deaf, 51.3% were bilingual, and 54.4% were not satisfied with the medical care. There was a statistically significant difference between the level of satisfaction of the volunteers with the health care received in relation to the different health professionals who attended them (p< 0.05). Conclusion: most of the population was not satisfied with medical care, although this service was most sought. The type of communication used by the professionals and the presence of an interpreter were not effective. It is necessary to implement strategies to ensure accessibility and comprehensive careto this population. RESUMO Objetivo: investigar a satisfação de surdos em relação aos Serviços de Saúde, caracterizar a amostra quanto aos aspectos sóciodemográficos, sócioeconômicos e auto declaração da surdez e verificar se há associação entre satisfação com o atendimento, comunicação, profissional, e autodeclaração da surdez. Métodos: estudo observacional transversal realizado com questionário semiestruturado, em 74 adultos surdos. Foi realizada análise descritiva da caracterização da amostra e do atendimento e análise de associação entre satisfação, profissional, autodeclaração, serviço utilizado e escolaridade, por meio do Qui-quadrado. O nível de significância adotado foi de 5%. Resultados: a amostra foi composta, prioritariamente, por mulheres (66,2%) entre 18 e 28 anos (46,5%), de classe D (51,3%), com ensino médio (56,76%), auxiliares de escritório ou “digitadoras” (90%). 63,5%, se autodeclara “surdo”, 51,3% é bilíngue e 54,4% não está satisfeito com o atendimento médico. Houve diferença estatisticamente significante entre a satisfação e o atendimento dos diversos profissionais (p< 0,05). Conclusão: a maioria da população não se mostrou satisfeita com o atendimento médico, embora este tenha sido o mais procurado. A modalidade de comunicação utilizada pelos profissionais e a presença de intérprete não foram efetivas. É necessária a implementação de estratégias para garantir a acessibilidade e integralidade à saúde dessa população.

A “tragédia da Piedade”, o grande drama social republicano do início do século XX, que envolveu toda a sociedade brasileira de 1909, é o objeto desta pesquisa, cujo tema é o valor da honra. O cenário dessa tragédia foi o Rio de Janeiro, então capital do Brasil, e seu palco, o subúrbio da Piedade. O escritor Euclides da Cunha foi uma das dramatis personae do acontecimento, ao lado da sua esposa, Anna Emília, e do amante dela, Dilermando de Assis. Euclides, famoso pelo seu opus magnum Os sertões : campanha de Canudos, diante da traição de sua esposa, tomou a decisão de lavar sua honra com sangue. No duelo, contudo, foi morto por Dilermando. O homicídio do marido traído pelo amante de sua esposa tornou-se um escândalo, suscitando, como problema de pesquisa, o custo social do valor da honra. This article is about the ‘tragédia da Piedade’. Its main theme is honor and its social costs. It analyzes a criminal case which happened in Brazil at the beginning of the twentieth century, when the Republican regime was just finishing its first decade of existence. The dramatis personae of the tragedy involved Brazil’s most famous writer, Euclides da Cunha, the renowned author of Os Sertões: Campanha de Canudos. He lived in a society which was changing, and where patriarchal tradition and State Law coexisted side-by-side. When Euclides’ wife, Anna Emília, betrayed him, he reacted as a traditional male would have done. He decided to wash his honor with blood. But, in the showdown with the young lover of his wife, Dilermando de Assis, he was killed by his opponent, unleashing a full-blown social drama which shook the very foundations of the whole society of Rio de Janeiro, the capital of Brazil at the time. The case, which reached its peak in a neighborhood called ‘Piedade’, became something of a national scandal about honor and shame, guilt and vengeance.

Made available in DSpace on 2019-10-06T16:08:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-12-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) β-thalassemia is a worldwide distributed monogenic red cell disorder, characterized by an absent or reduced beta globin chain synthesis. The unbalance of alpha-gamma chain and the presence of pathological free iron promote severe oxidative damage, playing crucial a role in erythrocyte hemolysis, exacerbating ineffective erythropoiesis and decreasing the lifespan of red blood cells (RBC). Catalase, glutathione peroxidase and peroxiredoxins act together to protect RBCs from hydrogen peroxide insult. Among them, peroxiredoxins stand out for their overall abundance and reactivity. In RBCs, Prdx2 is the third most abundant protein, although Prdxs 1 and 6 isoforms are also found in lower amounts. Despite the importance of these enzymes, Prdx1 and Prdx2 may have their peroxidase activity inactivated by hyperoxidation at high hydroperoxide concentrations, which also promotes the molecular chaperone activity of these proteins. Some studies have demonstrated the importance of Prdx1 and Prdx2 for the development and maintenance of erythrocytes in hemolytic anemia. Now, we performed a global analysis comparatively evaluating the expression profile of several antioxidant enzymes and their physiological reducing agents in patients with beta thalassemia intermedia (BTI) and healthy individuals. Furthermore, increased levels of ROS were observed not only in RBC, but also in neutrophils and mononuclear cells of BTI patients. The level of transcripts and the protein content of Prx1 were increased in reticulocyte and RBCs of BTI patients and the protein content was also found to be higher when compared to beta thalassemia major (BTM), suggesting that this peroxidase could cooperate with Prx2 in the removal of H 2 O 2 . Furthermore, Prdx2 production is highly increased in RBCs of BTM patients that present high amounts of hyperoxidized species. A significant increase in the content of Trx1, Srx1 and Sod1 in RBCs of BTI patients suggested protective roles for these enzymes in BTI patients. Finally, the upregulation of Nrf2 and Keap1 transcription factors found in BTI patients may be involved in the regulation of the antioxidant enzymes analyzed in this work. Universidade Federal de São Carlos (UFSCar) Departamento de Genética e Evolução Universidade de Campinas (UNICAMP) Departamento de Genética Evolução e Bioagentes Universidade Federal de Pernambuco (UFPE) Departamento de Genética Fundação de Hematologia e Hemoterapia do estado de Pernambuco (HEMOPE) Hemocentro da Universidade de Campinas (UNICAMP) Universidade Estadual Paulista (UNESP) Campus Litoral Paulista Universidade de São Paulo (USP) Departamento de Genética Biologia Evolutiva Universidade Estadual Paulista (UNESP) Campus Litoral Paulista FAPESP: 2011/50358-3

This article aims to demonstrate the contribution of the use of the Neurolinguistic Programming (NLP) tools to improve the results of the goals in an agency of a banking institution in Curitiba. This is a case study because it sought to measure the results of applying the NLP tools in the sales of the operational and commercial areas of a bank branch. It was also characterized as a descriptive and exploratory research, since it compared the results obtained in the sales approach, before and after the application of NLP tools. The professionals participating in the study were qualified and the tools were used within 30 days. It was then measured the result of that period and compared to the average of the results of the last six months to identify the evolution of the results obtained. Due to the depth of the techniques in relation to the conscious and unconscious of the people, it was evident that the time of learning and application of the techniques must be more extended than in fact was executed for the development of the article, With the expected results in sales.

Hydroxyurea (HU), or hydroxycarbamide, is used for the treatment of some myeloproliferative and neoplastic diseases, and is currently the only drug approved by the FDA for use in sickle cell disease (SCD). Despite the relative success of HU therapy for SCD, a genetic disorder of the hemoglobin β chain that results in red-cell sickling, hemolysis, vascular inflammation and recurrent vasoocclusion, the exact mechanisms by which HU actuates remain unclear. We hypothesized that HU may modulate endothelial angiogenic processes, with important consequences for vascular inflammation. The effects of HU (50-200 μM; 17-24 h) on endothelial cell functions associated with key steps of angiogenesis were evaluated using human umbilical vein endothelial cell (HUVEC) cultures. Expression profiles of the HIF1A gene and the miRNAs 221 and 222, involved in endothelial function, were also determined in HUVECs following HU administration and the direct in vivo antiangiogenic effects of HU were assessed using a mouse Matrigel-plug neovascularization assay. Following incubation with HU, HUVECs exhibited high cell viability, but displayed a significant 75% inhibition in the rate of capillary-like-structure formation, and significant decreases in proliferative and invasive capacities. Furthermore, HU significantly decreased HIF1A expression, and induced the expression of miRNA 221, while downregulating miRNA 222. In vivo, HU reduced vascular endothelial growth factor (VEGF)-induced vascular development in Matrigel implants over 7 days. Findings indicate that HU is able to inhibit vessel assembly, a crucial angiogenic process, both in vitro and in vivo, and suggest that some of HU's therapeutic effects may occur through novel vascular mechanisms.

Hereditary persistence of fetal hemoglobin deletion type-2 (HPFH-2) and Sicilian-δβ-thalassemia are conditions described as large deletions of the human β-like globin cluster, with absent β-globin chains and a compensatory variable increase in γ-globin. HPFH, in general, may be distinguished from DB-Thalassemia by higher fetal hemoglobin (HbF) levels, absence of anemia and hypochromic and microcytic erythrocytes. MicroRNAs (miRNAs) regulate a range of cellular processes including erythropoiesis and regulation of transcription factors such as the BCL11A and SOX6 genes, which are related to the regulation of γ-globin expression. In this report, a possible association among the overexpression of miRNAs and the expression of the γ-globin gene was analyzed in these two conditions. Forty-nine differentially expressed miRNAs were identified by microarrays in CD34+-derived erythroid cells of two subjects heterozygous for Sicilian-δβ-thalassemia, 2 for HPFH-2 and 3 for controls after 13 days of culture. Some of these miRNAs may participate in γ-globin gene regulation and red blood cell function. The BCL11A gene was found to be potentially targeted by 12 miRNAs that were up-regulated in HPFH-2 or in DB-Thal. A down-regulation of BCL11A gene expression in HPFH-2 was verified by quantitative polymerase chain reaction. These data suggest an important action for miRNA that may partially explain the phenotypic differences between HPFH-2 and Sicilian δβ-thalassemia and the increased expression of γ-globin in these conditions.

Sickle cell disease (SCD) is a complex disease that is characterized by the polymerization of deoxyhemoglobin S, altered red blood cell membrane biology, endothelial activation, hemolysis, a procoagulant state, acute and chronic inflammation, and vaso-occlusion. Among the physiological changes that occur during pregnancy, oxygen is consumed by fetal growth, and pregnant women with SCD are more frequently exposed to low oxygen levels. This might lead to red blood cells sickling, and, consequently, to vaso-occlusion. The mechanisms by which SCD affects placental physiology are largely unknown, and chronic inflammation might be involved in this process. This study aimed to evaluate the gene expression profile of inflammatory response mediators in the placentas of pregnant women with sickle cell cell anemia (HbSS) and hemoglobinopathy SC (HbSC). Our results show differences in a number of these genes. For the HbSS group, when compared to the control group, the following genes showed differential expression: IL1RAP (2.76-fold), BCL6 (4.49-fold), CXCL10 (−2.12-fold), CXCR1 (−3.66-fold), and C3 (−2.0-fold). On the other hand, the HbSC group presented differential expressions of the following genes, when compared to the control group: IL1RAP (4.33-fold), CXCL1 (3.05-fold), BCL6 (4.13-fold), CXCL10 (−3.32-fold), C3 (−2.0-fold), and TLR3 (2.38-fold). Taken together, these data strongly suggest a differential expression of several inflammatory genes in both SCD (HbSS and HbSC), indicating that the placenta might become an environment with hypoxia, and increased inflammation, which could lead to improper placental development.

Made available in DSpace on 2014-06-11T19:29:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-01-28Bitstream added on 2014-06-13T20:20:15Z : No. of bitstreams: 1 santana_rf_me_arafcl.pdf: 1280059 bytes, checksum: 3917f040b16325ff31d971c16acba2f7 (MD5) A dificuldade no ensino da matemática, reiteradamente apontada pelos professores, é evidenciada pelos baixos índices de aprendizagem dos alunos, segundo os dados divulgados pelas avaliações externas que recaem sobre a educação escolar brasileira. Investigar a organização das aulas, para então discutir sobre uma proposição teórica e metodológica para as mesmas, é o objetivo central desta dissertação. Diante do exposto, e por entender que uma correta organização e desenvolvimento dos conceitos pode colaborar para a aprendizagem destes, visamos desenvolver uma proposição metodológica para a aula coerente com o materialismo histórico dialético, a partir de atividade que desenvolva conceitos e esteja fundamentada nas categorias atividade mediada1 e mediação dialética e pedagógica2, as quais serão estudadas no decorrer dos capítulos. A intenção desta investigação é a de colaborar com as discussões atuais sobre o ensino e aprendizagem de matemática em sala de aula, elaborando, a partir das categorias citadas, um corpo teórico que envolva a organização metodológica do conceito, o seu desenvolvimento e a sua avaliação como alternativa metodológica que proporciona aos alunos a elaboração dos conceitos matemáticos por meio da superação The difficulty in teaching mathematics, repeatedly pointed out by teachers, is evidenced by low levels of students’ learning, according to data released through the ratings that are attributed to Brazilian education. To investigate classes organization and then discuss about a theoretical and methodological proposition for these classes is the fundamental objective of this essay. Considering the above and our belief (that the correct organization and development of concepts can help their learning), we aim to develop a methodological proposition for the math class as an educational activity that develops concepts from the categories mediated activity1 and dialectical and educational mediation2. The intent of this research is to collaborate with the current discussions about teaching and learning mathematics preparing, from the categories mentioned above, a theoretical methodology involving the organization of the concept, its development and its evaluation as a methodological alternative, which provides students with the creation of mathematical concepts by overcoming their difficulties

Introduction: Mouse models have allowed a wider investigation of the inflammatory process in Sickle cell disease (SCD). Previous studies have demonstrated that neutrophil recruitment inresponse to LPS is upregulated in Berkeley sickle mice and the development of acute LPS-induced airway inflammation is associated with enhanced expression of cytokines, chemokines and MMPs expression in lungs. MicroRNAs (miRNAs) belong to a regulatory class of RNAs with great importance in several biological processes, including the development and fate of immune cells, and in both innate and adaptive immune responses, whereby strong evidence links miRNAs expression to signalling pathways and receptors. Some miRNAs are positive regulators of inflammation and others are involved in the negative-feedback regulation of inflammatory processes. A growing amount of evidence indicates that these molecules are involved in various lung diseases. However, the expression profile of miRNAs in lung injury in SCD have not been investigated. Since pulmonary complications are an important cause of morbidity and mortality in SCD, the aim of this study was to determine whether miRNAs implicated in LPS-mediated inflammation are involved in neutrophil recruitment and inflammation in lung injury in SCD using the sickle mice model. Methods: Acute lung inflammation and injury was induced in wild- type (WT, C57BL/6, n=5) and Berkeley SCD mice (n=5) by intranasal administration of LPS (50 ml of 250 mg/ml). The vehicle group received a similar volume of sterile PBS. BAL was performed 4h after LPS challenge. qPCR analysis was used to examine gene expression and ELISA was used to determine protein production. For miRNAs expression analysis, extraction of lung RNA was performed using mirVanaTM miRNA Isolation kit and TaqManTM MicroRNA Reverse Transcription.The expression level of miRNAs was determined using the Taqman miRNA expression assay by qPCR with U6 snRNA as an endogenous control. Results: Levels of miR-15a-5p, miR-16-5p, miR-26a-5p and miR-98-5p in lung tissue increased almost twofold at 4h after LPS challenge in SS compared with SS PBS group (2.9, 3.7; 2.3; 2.9-fold change, respectively). The expressions of miR-106a-5p, miR-146a-5p, miR-146b-5p, and miR-181a-5p were not altered (2.0; 1.2; 1.55; 1.4-fold change, respectively). However, the expressions of miR-203a-3p and miR-223-3p were significantly up-regulated compared to animals from the PBS group (4.3 and 8.8 fold change, respectively, p Conclusion: We have shown that theLPS-induced inflammatory response in SCD transgenic mice is associated with increases in miR-203a-3p and miR-223-3p expression in the mouse lung; suggesting a role of the miRNAs in the regulation of inflammatory mediator production, since these augmentations are associated with the increases in cytokines and chemokines. Furthermore, miR-146a-5p, a negative regulator of inflammatory gene expression, is not upregulated in SS mice and could contribute to the exaggerated inflammation observed in these animals. These data indicate that alterations in the miRNA expression may be associated with the inflammatory state in SCD. Financial Support: FAPESP Disclosures No relevant conflicts of interest to declare.

Erythropoiesis is controlled by a complex interplay of signaling pathways, key transcription factors (TFs), as well as microRNAs (miRs). Although studies have identified miRs and TFs that regulate erythroid commitment and differentiation, the mechanism by which these molecules contribute to erythroid disorders, such as beta thalassemia (BT), is unclear. The aim of this study was to investigate miRs and TFs expression during in vitro erythroid differentiation in normal and ineffective erythropoiesis. Peripheral blood CD34+ cells were isolated from a transfusion-independent BT intermedia patient, homozygous for HBB IVS-I-6 (T→C), and cultured in a liquid system stimulating erythropoiesis. Cells were collected on day 10 for quantification of 84 mature miRs using a miRNA PCR Array platform. To minimize interindividual genetic variability, a control sample was collected from a healthy sibling (CON). For validation, CD34+ cells from 4 other BT patients and 4 healthy subjects were cultured and collected on days 7, 10, and 13 to measure miRs and TFs expressions during erythroid differentiation. Results: Data analysis from the miRNA PCR Array showed deregulated expression of 13 miRs in BT compared to CON: miRs 150-5p and 7-5p were decreased (fold change: -3.26 and -2.80) and miRs 32-5p, 96-5p, 155-5p, 146a-5p, 210, 125b-5p, 99a-5p, 125a-5p, 124-3p, 196b-5p, and 130a-3p were increased (fold change: 2.04; 3.72; 3.04; 2.27; 8.42; 2.82; 3.13; 2.48; 5.14; 2.00, and 3.07). The expression profile of ten miRs was successfully validated with the samples on day 10 of BT cultures. In addition, miRs 96-5p, 155-5p, and 210 remained increased on day 13 (P Conclusion: Our data report associations between differentially expressed miRNAs and their possible targets during in vitro beta thalassemic erythroid differentiation. These results corroborate the importance of miRNAs in the hematopoietic process and highlight that further studies of the interaction between TFs and miRs may contribute to a better understanding of the pathophysiology of ineffective erythropoiesis. Support by FAPESP and CNPq Disclosures No relevant conflicts of interest to declare.

BACKGROUND: Deep vein thrombosis (DVT) is multi-causal disease associated to a high morbi-mortality due to complications as pulmonary embolism and post-thrombotic syndrome. The identification of factors enrolled on the disease is important to better understand its pathophysiology. As the role of the platelets on DVT is still not completely defined, we previously performed a proteomic study on platelets from 3 DVT patients and compared to results obtained from 1 sibling and 1 neighbor for each patient in order to minimize the genetic and environmental interferences. These high-selected patients presented spontaneous and recurrent episodes of proximal DVT and mentioned a familiar history of thrombotic diseases. METHODS: platelets were washed, lysed, and their proteins were alkylated, reduced, precipitated with acetone and hydrolyzed by trypsin. 100mg of peptides were then separated by hydrophobicity using HPLC, and 8 fractions were obtained and directed to the LTQ-Orbitrap mass spectrometer. The proteins search was performed by Sorcerer-SEQUEST. In this analysis, sorbitol dehydrogenase (SORD) was present only in patients and absent in siblings and neighbors. In order to evaluate the SORD gene expression, qPCR was performed on platelets RNA from the same individuals applying specific primers after the evaluation of a minimal leukocyte contamination with specific primers to CD45. The influence of SORD on platelets adhesion was evaluated with the administration of a direct inhibitor (fidarestat) to 12 mice C57Bl6 by gavage at 10mg/kg and the results were compared to that obtained from 12 animals gavaged in a solution of 5% gum arabic. To the adhesion test, blood was obtained from cava veins in 3.8% sodium citrate and centrifuged at 100G/10’. Platelets were collected, pelleted and washed in Krebs buffer. Platelets at 1.2 x 108/μL were added to 96 wells plates pre-covered with fibrinogen 50μg/mL and their adhesion properties were evaluated with and without stimulation with 50mU of thrombin after incubation at 37°C for 30’. The percentage of adherent cells was calculated by comparing acid phosphatase activity to that of a standard curve with known numbers of platelets (0 – 100%) from the same group at 405nm. On the qPCR, the analyses were performed with fold change, and we considered significant results lower then 0.5x and higher then 1.5x. The Mann-Whitney test was applied to analyze the adhesion results, and P RESULTS: On qPCR, the gene expression of SORD was increased in patients (1x) in comparison to siblings (0.51x), but similar to neighbors (0.77x). The adhesion of platelets unstimulated was similar between both groups of animals (Median fidarestat= 41.6; 2.5-91.6; SD=25.9; median controls = 45; 9.8-91.3; SD=22.6; P=0.69), but after stimulation with thrombin, platelets from treated animals presented increased adhesion (median = 74.4; 42.9-98.8; SD=24) when compared to controls (median = 43.9; 13.6-48.2; SD=12.1; P=0.0148). CONCLUSIONS: SORD was present only in patients with DVT and absent in healthy siblings and neighbors on the proteomic analysis. The increase of SORD gene expression in patients in comparison to all the controls can corroborate the involvement of this protein on DVT physiology. The inhibition of this protein with fidarestat promoted the increase of platelets adhesion to plates covered with fibrinogen only after stimulation with thrombin. Our hypothesis is that SORD may be a protective protein based on the increase of the gene expression, protein levels and adherent properties after thrombogenic stimuli that could be an attempt to decrease the platelet function and minimize the damage. Disclosures No relevant conflicts of interest to declare.

Introduction: Pregnant women with sickle cell disease (SCD) are at higher risk of obstetrical complications, such as preeclampsia and preterm delivery. The placental circulation is susceptible to vaso-occlusion, which may contribute to fibrosis, necrosis and infarction of the placental villi. Restricted uteroplacental circulation is associated with low birth weight and fetal growth restriction. The mechanisms by which SCD affects placental physiology are largely unknown, and the chronic inflammatory state may be involved in this pathological process. The aim of this study was to evaluate the expression of mediators of inflammatory response in the placenta of pregnant women with SCD. Methods: This is an ongoing case control study that has recruited 4 pregnant patients with hemoglobin SC disease and 4 healthy control pregnant women paired for gestational age and route of delivery. Placental tissue was collected for total RNA extraction and synthesis of cDNA. Samples were run on a qPCR Array (Inflammatory Response and Autoimmunity RT2 Profiler, Qiagen), which includes 84 genes. Gene expression levels were normalized to housekeeping genes (ACTB, B2M, and RPLP0), and will be further validated by RT-qPCR. Results: Two patients underwent cesarean delivery and one patient delivered preterm. The analysis of placental gene expression showed that 16 genes were upregulated. The top 5 most upregulated genes were CCL24 (18.1 fold), TLR9 (8.6 fold), inducible nitric oxide synthase NOS2 (6.8 fold), IL17A (5.4 fold), and CCL16 (4.8 fold). Upregulated genes also included other chemokines and their receptors (CCL21, CCL22, CXCL3, and CCR7), interleukins (IL1RAP, IL5, and IL9), another toll-like receptor (TLR3), transcription factor BCL6, CD40 ligand (CD40LG), and kininogen 1 (KNG1). Interestingly, the two genes found to be downregulated were CRP (C reactive protein) and IFNG (interferon gamma). Conclusion: This is the first study of differential gene expression in placenta of patients with SCD. Despite patients with hemoglobin SC disease being clinically less severe than homozygous sickle cell anemia, we have found that placental tissue in this subset of patients displays increased expression of several genes involved in inflammatory response. Our data support local inflammatory changes in the placenta of these patients that might explain the development of complications in SCD pregnancies. Although the association between obstetric complications and SCD has been long described, our data should contribute to better define the mechanisms affecting the placental physiology that lead to complications, such as preeclampsia. Disclosures No relevant conflicts of interest to declare.

Introduction Hereditary Persistence of Fetal Hemoglobin (HPFH) and δβ-thalassemia are genetic disorders characterized by elevated levels of fetal hemoglobin (HbF) in adulthood. Deletions of variable sizes, and at different positions, that involve the β-globin gene complex on chromosome 11 are associated with these disorders. The distinction between these conditions is made based on clinical and hematological data. Deletional HPFH includes a wide range of conditions, but typically it is characterized in heterozygotes by levels of HbF of 15% to 30% with normal red blood cell indices, while heterozygotes for δβ-thalassemia tend to have elevated levels of HbF that are lower (5% to 20%) and accompanied by mild anemia with hypochromic, microcytic red blood cell indices. MicroRNAs (miRNAs) have been described to have a possible role in globin switching and can modulate transcriptional erythroid-specific regulators. To the best of our knowledge miRNAs have not been analyzed in HPFH and δβ-thalassemia. The aim of this study was to investigate the miRNAs expression profile and possible post-transcriptional role of these molecules in relation to the lack of normal suppression of γ genes in these genetic disorders. Methods CD34(+)-derived erythroid cells from two HPFH-2 individuals and two δβ-thalassaemia Sicilian type patients (DB) and healthy controls (CTRL) were cultured for 13 days to examine the expression profile of miRNAs. The miRNAs were hybridized using an Agilent miRNA microarray platform and the profiles were obtained through bioinformatics data analysis using GeneSpring software. qPCR analysis was used to validate the miRNA expression (TaqMan® miRNAs assays) and to quantify gene expression of 19 transcription factors. Different databases, such as miRBase, TargetScan, microRNA.org and BioGPS were used to determine the predicted targets of miRNAs data found. Results Six miRNAs were up-regulated in HPFH and in DB compared to CTRL: miR-146b-5p, miR-181a, miR-342-3p, miR-362-3p, miR-362-5p and miR-365. Five miRNAs were up-regulated in HPFH, compared to DB: miR-223, miR-630, miR-638, miR-1246 and miR-1290. Nine miRNAs were up-regulated in DB compared to HPFH: miR-10a, miR-21*, miR-33b*, miR-96, miR-128, miR-194, miR-210, miR-424 and miR-1275. The ALK4 and the GATA2 mRNAs were significantly up-regulated in HPFH, compared to DB. The BCL11A and the SH3BGRL2 mRNAs were significantly down-regulated in HPFH compared to DB. In silico analysis and the literature show that several miRNAs have targets related to HbF production or erythropoiesis. These include miR-10a, miR-33b*, miR-96, miR-128, miR-210, miR-223, miR-342-3p, miR-362-3p, miR-424 and miR-630. Conclusion The comparison of miRNA and transcription factor profiles suggests differences in the expressions of several miRNAs that may influence γ-globin gene expression. These data may contribute to understanding the phenotypic differences found between deletional HPFH and δβ-thalassemia. Financial support by FAPESP and CNPq/INCTS. Disclosures: No relevant conflicts of interest to declare.

Abstract 3201 During erythroid differentiation, the expression profile of erythroid-specific genes follows a precise timing that is controlled by a complex network of transcription factors (TFs) and microRNAs (miRNAs) that regulate the expression of a set of target genes. The aim of this study was to analyze the expression profile in sickle cell anemia (SCA) and β-thalassemia (BT) patients of the TFs GATA-1, TAL-1, NFE-2, LDB1, LMO2, EKLF and of the miRNAs 24, 144, 155, 210, 221, 222, 223 and 451, which all of have been previously described in normal erythropoiesis. Reticulocytes were separated from peripheral blood samples of subjects with normal hematological data (n≥8), from untransfused BT IVS-I-6 (T→C) homozygous patients (n=9) and from SCA patients (n=8), without HU therapy. Spherocytosis patient samples were included in the results analysis. Extraction of RNA and miRNA, transcription to cDNAs and qRT-PCR were performed to analyze TFs and miRNAs expressions. In BT patients, statistical analyzes showed that the expression levels of the TFs GATA-1, TAL-1 and NFE-2 were significantly higher (p Support by FAPESP, CNPq and INCTS Disclosures: No relevant conflicts of interest to declare.

Abstract 3195 The hereditary persistence of fetal hemoglobin (HPFH) is the consequence of impaired switching in adult life, which results in the continued expression of gamma globin gene. The Brazilian HPFH type is characterized by a C → G substitution at the –195 position of the A gamma globin gene promoter, and associated with HbF levels ranging from 6 to 16% in the heterozygote state. This study was undertaken to identify genes that may be involved in hemoglobin switching and/or maintenance of elevated HbF levels in Brazilian HPHF subjects. The Suppressive Subtractive Hybridization libraries were constructed using a pool of RNA extracted from reticulocytes of peripheral blood in individuals with normal hematological data and from subjects with Brazilian and 55 and 57 overexpressed genes were identified in the normal and Brazilian HPFH library, respectively; findings were validated by qRT-PCR and Western blotting. One transcription factor identified was FOXO3a, whose expression was increased in Brazilian HPFH subjects, compared to control subjects. Moreover, the non-phosphorylated Foxo3a protein that interacts with DNA, was detected only in Brazilian HPFH when analyzed by Western blotting. FOXO3a binds to PAX1 promoter, a transcription factor whose activity was higher in Brazilian HPFH. The FOXO3a/PAX1 complex may be important in the mechanism responsible for increasing HbF levels in this genetic disorder. Another gene analyzed was KLF1, a transcription factor known as a regulator of switching from the gamma to beta globin gene. This gene was underexpressed in the reticulocytes of HPFH subjects. Klf1 protein activity in erythroid cells of healthy donors was also increased compared to Brazilian HPFH, when analyzed by DNA/protein array. Results suggest that the decreased KLF1 levels in Brazilian HPFH favors the interaction between the A gamma globin gene and the Locus Control Region, agreeing with previous studies that demonstrated that knockdown of KLF1 in adult erythroid progenitors favors the formation of complexes that bind to the gamma globin gene promoter. MIER1 expression was also found to be decreased in Brazilian HPFH reticulocytes, compared to controls. The MIER1 gene is able to recruit chromatin remodeling-enzymes leading, to the formation of heterochromatin and, consequently, silencing genes. This MIER1 may be an important gene in gamma to beta globin gene switching, where it could help in the maintenance of a closed chromatin structure in the gamma globin gene in individuals with low HbF levels. The expression of the HOOK3 gene was decreased in Brazilian HPFH reticulocytes, compared to controls. The HOOK3 encodes a protein that interacts with a GTPase protein, stimulating INF-gamma, responsible for the phosphorylation of p65/p50 and RXR beta like proteins which regulate the transcription of the beta globin gene. The reduced beta globin gene expression in Brazilian HPFH resulting from the reactivation of gamma globin gene may be responsible for a decrease in the HOOK3 expression. These results suggest that, in the HPFH Brazilian type, the FOXO3a/PAX1 complex could contribute to the continued expression of the A gamma globin gene. Additionally, some cellular modifications, such as low Klf1 activity and decreased HOOK3 and MIER1 expression, could participate in the maintenance of HbF levels. These genes could be important in therapeutic approaches for the development of new HbF induction agents for the hemoglobinopathies. Support by FAPESP, CNPq.and INCTS Disclosures: No relevant conflicts of interest to declare.