Orientador: Antonio Sergio Ramalho / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-07-19T08:31:44Z (GMT). No. of bitstreams: 1 Calusni_AnaLuciaRoscani_M.pdf: 1715301 bytes, checksum: acd3974eacbc813ea7e1416eeec145b7 (MD5) Previous issue date: 1994 / Mestrado / Genetica / Mestre em Ciências Biológicas

Orientador: Sara T. Olalla Saad / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-07-19T12:14:28Z (GMT). No. of bitstreams: 1 Pranke_PatriciaHelenaLucas_M.pdf: 3795822 bytes, checksum: fba966483775191a766c241619c01e17 (MD5) Previous issue date: 1994 / Resumo: No presente trabalho foram analisados 10 pacientes com EIH e seus familiares através do estudo das proteínas totais em SDS-P AGE (Sistema Fairbanks 3,5-17% e Sistema Laemmli 12%), estudo da auto-associação de heterodímeros de espectrina e digestão tríptica da espectrina. Seis destas famílias apresentaram defeito na auto-associação dos heterodímeros de espectrina, sendo dois casos de Sp a 1150-46, três famílias com Sp a 1165e uma família com uma ß-espectrina encurtada, conforme evidenciado por técnica de westem-blotting com anticorpo anti-ß espectrina. Numa das famílias com Sp a 1165, um dos pacientes apresentou também uma ß-espectrina alongada, mas esta associação não determinou maior gravidade da doença ou aumento dos dímeros de espectrina. Uma família e um paciente apresentaram deficiência de proteína 4.1 e em dois casos não foi possível detectar, pela metodologia empregada, o defeito proteico. Nos pacientes Sp a 1165 kDa a pesquisa de heteroduplex após a amplificação do exon 4 do DNA genômico da cadeia alfa da espectrina demonstrou o aparecimento de uma banda com migração retardada nas três famílias estudadas sugerindo a duplicação do codon TTG. Em uma paciente Sp a 1150-46 kDa o sequenciamento do exon 6 da cadeia a-sp mostrou uma mutação de ponto CTG leu -+ CCG pro em um alelo no codon 260. Este é o primeiro estudo das alterações proteicas da ElH na população brasileira. A ß-espectrina alongada observada neste trabalho é o segundo caso descrito na literatura enquanto que a ß-espectrina "truncada" é, provavelmente, a menor ß-sp já descrita. O estudo da auto-associação dos heterodíineros de espectrina foi também levado a efeito em 26 pacientes com doenças cujo aparecimento de eliptócitos no esfregaço de sangue periférico era freqüente (7 LMC, 6 mielofibrose, 5 anemia ferropriva, 3 anemia megaloblástica e 5 anemia falciforme). Destes, um paciente com LMC apresentou aumento de dímeros de espectrina sugerindo, neste caso, um prejuízo na auto-associação de heterodímeros de espectrina / Abstract: The aim of this work was to study the protein defects in Hereditary Elliptocytosis (HE) in a brazilian population and to analyse the self-association of spectrin in patients without HE whose elliptocytes are frequently observed. Red cell membrane of ten patients with HE and their rei atives were investigated by SDS-PAGE (Fairbanks' system 3,5-17% and Laemmli's system 12%), by non denaturing gels for spectrin heterodimer self-association study and by the tryptic digestion of spectrin. Six families presented a defect in the spectrin self-association: two had Sp a I/50-46, three with Sp a I/65 and one with a "truncated" ß-spectrin as observed by westem-blotting using anti-ß espectrin antibody. In one family with Sp a I/65, one patient presented also a elongated ß-espectrin but this association did not increase the amount of spectrin dimers or the clinical severity of the disease. One family and one patient presented a deficiency of protein 4.1 and in two families, the protein defect was not detected by the approach used in this study.In the Sp a I/65 kDa patients, the analysis of heteroduplex after amplification of exon 4 of the genomic DNA for the a-spectrin chain revealed the appearance of a slower migrating band in the three families studied suggesting the duplication of codon TTG. In one patient with Sp a I/50-46 kDa the sequencing of exon 6 of the a-sp chain showed a point mutation CTG leu - CCG pro in an allele in codon 260. As far as we know, this is the first study of red cell membrane in hereditary elliptocytosis in a brazilian population. The elongated ß -spectrin observed in this study is the second case described in the literature and the truncated ß-spectrin probably corresponds to the smaller ß-sp already described.The study of the spectrin heterodimer self-association was also performed in 26 patients with diseases in which elliptocytes are frequently observed in peripheral blood smears (7 CML, 6 myelofibrosis, 5 iron deficiency anemia, 3 megaloblastic anemia and 5 sickle cell disease). Only one patient with CML had increased IeveIs of the spectrin dimer, which suggested a defect in the spectrin heterodimer self-association / Mestrado / Mestre em Farmacologia

Varios fatores contribuem para o desenvolvimento da anemia, que constitui um dos mais graves problemas de saude publica. A anemia hipocromica microcitica e a forma mais comum em criancas e adolescentes. Dentre as causas desta anemia estao: a) deficiencia de ferro, que resulta de um longo periodo do balanco negativo do micronutriente e causa retardo no crescimento e comprometimento do desempenho cognitivo de criancas; b) contaminacao por chumbo (plumbismo) que tambem afeta o desenvolvimento das criancas, podendo ser agravada nos portadores de polimorfismo da enzima ALAD; c) hemoglobinopatias (hemoglobinas variantes e talassemias), anemias herdadas que afetam 7% da populacao mundial. Devido a alta prevalencia destas patologias, o presente trabalho teve como objetivo estudar um grupo de criancas de escolas publicas, identificando os fatores que contribuem para o desenvolvimento de anemias hipocromicas microciticas e estabelecer relacoes entre as caracteristicas laboratoriais das doencas. Participaram do estudo 427 criancas, com idade entre 6 a 9 anos, sendo 235 do sexo feminino e 192 do sexo masculino, alunos de Escolas Municipais e Estaduais, da zona norte da cidade de Ribeirao Preto-SP. Foram analisados: a) numero global de eritrocitos e leucocitos, concentracao de hemoglobina, hematocrito, indices hematimetricos e distribuicao da amplitude das celulas vermelhas (contador automatico Micros 45 . Horiba ABXR) e calculo do indice matematico RDWI; b) niveis plasmaticos de chumbo (espectrometro de massa com plasma indutivamente acoplado VG Plasmaquad PQIIR) e estudo das delecoes dos polimorfismos da enzima ALAD, por PCR; c) status ferrico pelos niveis de ferritina serica (imunoquimioluminescencia utilizando kit Ferritin Immulite . DPCR e equipamento Immulite 1 - DPCR), receptor de transferrina soluvel (ensaio imunoenzimatico, utilizando o kit Quantikine soluble transferrin receptor da R&D SystemsR e o leitor de microplacas de ELISA READER 210, modelo Microwell System Organon TeknikaR) e calculo do indice sTfR/log ferritina; d) analise das hemoglobinas por eletroforese em acetato de celulose, pH alcalino, por HPLC (sistema automatizado Variant II Bio-RadR e kit gÀ-talassemia Short Program) e PCR para a principal delecao de ¿- talassemias. Com base no criterio recomendado pela OMS para definir anemia (Hb menor que 11,5 g/dL), verificou-se que 75 (17,6%) criancas eram anemicas, sendo 33 (44%) portadoras de algum tipo de hemoglobinopatia, 29 (38,6%) com anemias de causa desconhecida e 13 (17,4%) com anemia por deficiencia de ferro. Das anemias, apenas 14 eram anemias hipocromicas microciticas, sendo que 10 (71,4%) eram algum tipo de hemoglobinopatia, 2 (14,2%) ADF e 2 (14,2%) de causa desconhecida. Na populacao estudada, a prevalencia de hemoglobinopatias foi de 16,6% , a saber: 11,6% com ¿-talassemia; 4% com aumento de Hb F; 3,5% com Hb AS; 2,8% com À-talassemia; 0,96% com ¿/À-talassemia e 0,24% com Hb AC. Os niveis de chumbo plasmatico, em todos os participantes do estudo, estavam dentro do recomendado pelo Center for Disease Control and Prevention (< 10 Êg/dL), nao havendo interferencia do metal na patogenese das anemias. Nao houve associacao entre os polimorfismos da ALAD-1 (ALAD1-1 e ALAD1-2) e os niveis de chumbo plasmatico. Anemia por deficiencia de ferro foi diagnosticada em 3% das criancas e DF em 6,1%, utilizando um cut off de 30 ng/mL para ferritina serica. Houve concordancia na identificacao de hemoglobinopatias utilizando as metodologias eletroforese de hemoglobina em acetato de celulose e HPLC, sendo que estas metodologias nao sao uteis para diagnosticar ¿-talassemia. Para identificar os portadores da delecao de ¿-talassemia (.¿3,7) e necessaria a utilizacao da análise molecular (PCR). A suspeita de Hb S/-talassemia identificada por HPLC deve ser confirmada por análise dos pais e/ou irmãos. A ferritina foi um bom parâmetro para identificar DF precocemente e útil para diferenciar os portadores de hemoglobinopatias dos portadores de DF e ADF. O índice sTfR/log da ferritina foi mais sensível do que o sTfR, na diferenciação de DF e talassemia. No diagnóstico das anemias hipocrômicas microcíticas é necessário analisar um conjunto de determinações, incluindo exame hematológico, status férrico, perfil eletroforético, em alguns casos incluindo avaliação dos familiares, e análise molecular das hemoglobinopatias. / Several factors contribute to the development of anemia, which constitutes one of the most serious problems in public health. The hypochromic microcytic anemia is the most common type in children and adolescents. Among the causes for this type of anemia are: a) iron deficiency, which results from a long period of negative balance of the micronutrient, causing delay in growth and compromising the cognitive performance of the children; b) contamination by lead (lead poisoning), which also affects the development of children, and may be aggravated in carriers of polymorphism of the enzyme ALAD; c) hemoglobinopathies (variants hemoglobin and thalassemia), inherited anemia that affects 7% of the world population. Due to the high prevalence of these pathologies, the present study aimed at studying a group of children from public schools, identifying the factors that contribute to the development of hypochromic microcytic anemia and establishing relations between the laboratorial characteristics of the diseases. The study had the participation of 427 children, aged between 6 and 9 years old, being 235 female and 192 male students from Municipal and State Schools in the north area of Ribeirao Preto-SP. It analyzed: a) number of erythrocytes and leucocytes, hemoglobin concentration, hematocrit, red cell indices and red cell distribution width (automatic counter Micros 45 . Horiba ABXR) and calculation of the mathematical index RDWI; b) plasma lead levels (inductively coupled plasma mass spectrometer VG PlasmaQuad PQIIR) and study of the deletions of the polymorphisms of the enzyme ALAD, by PCR; c) iron status by serum ferritin levels (immunochemiluminescence using the kit Ferritin Immulite . DPCR and the equipment Immulite 1 - DPCR), soluble transferrin receptor (enzyme immune assay, using the kit Quantikine soluble transferrin receptor of R&D SystemsR and the microplate reader ELISA READER 210, model Microwell System Organon TeknikaR) and calculation of the sTfR/log ferritin index; d) hemoglobin analysis by electrophoresis on cellulose acetate at alkaline pH, HPLC (automated system Variant II Bio-RadR and the kit gÀ-thalassemia Short Program) and PCR for the main deletion of ¿-thalassemias. Based on the WHO criteria by to define anemia (Hb under 11.5 g/dL), it was verified that 75 (17.6%) children were anemic, being 33 (44%) with hemoglobinopathy, 29 (38.6%) with anemia of unknown causes and 13 (17.4%) with iron deficiency anemia. Among the anemias, only 14 were hypochromic microcytic, 10 (71.4%) being some sort of hemoglobinopathy, 2 (14.2%) due to iron deficiency and 2 (14.2%) due to unknown causes. In the studied population, the prevalence of hemoglobinopathies was 16.6%, namely: 11.6% with ¿-thalassemia; 4% with Hb F elevated; 3.5% with Hb AS; 2.8% with À- thalassemia; 0.96% with ¿/À-thalassemia and 0.24% with Hb AC. The plasma lead levels, in all participants of the study, were within the levels recommended by the Center for Disease Control and Prevention (< 10 Êg/dL), without the interference of the metal in the pathogenesis of the anemias. There was no significant association between the polymorphisms of the ALAD-1 (ALAD1-1 and ALAD1-2) and the plasma lead levels. Iron deficiency anemia was diagnosed in 3% of the children and ID in 6.1%, using a cutoff of 30 ng/mL for serum ferritin. There was agreement in the identification of hemoglobinopathies using the methodologies electrophoresis of hemoglobin in cellulose acetate and the HPLC, as these methodologies are not useful to diagnose ¿-thalassemia. In order to identify the carriers of ¿-thalassemia gene deletion (.¿3,7) it is necessary to use the molecular analysis (PCR). The suspicion of Hb S/À-thalassemia identified by HPLC must be confirmed through the analysis iv of the parents and/or siblings. The ferritin was a good parameter to identify ID early and useful to differ the carriers of hemoglobinopathies of the carriers of ID and IDA. The sTfR/log ferritin level was more sensitive than the sTfR, in the differentiation of ID and thalassemia. In the diagnosis of the hypochromic microcytic anemias, it is necessary to analyze a set of determinations, including hematological exam, iron status, electrophoretic profile, in some cases including relatives, and molecular analysis of the hemoglobinopathies.

Introducción: El plomo inhibe la síntesis del grupo hemo y en consecuencia de la hemoglobina. Objetivos: Evaluar la intoxicación plúmbica crónica y su relación con problemas de anemia en trabajadores de Siderperu. Población muestral: 50 trabajadores. Determinación de la plumbemia: Espectroscopia de absorción atómica con horno de grafito. Análisis de hemoglobina: colorimétrico para la determinación de hemoglobina como cianuro de hemiglobina en sangre. Resultados: En la correlación de los niveles de plumbemia y el tiempo de exposición de los trabajadores de Siderperu se muestra que no existe una correlación significativa entre la concentración de plomo en la sangre y el tiempo de exposición de los trabajadores. Respecto a la correlación de los niveles de plumbemia y el área de trabajo de los trabajadores el 34 % de la población de estudio labora en el área de proceso para barras mineras, el 18 % labora en el proceso para fierro de construcción, el 16 % en la planta de acero, el 14 % en talleres guiados, el 12 % en zona de estandarizado y el 6 % en bobinas de importación. El 6 % de los trabajadores tienen valores de plumbemia que requieren pasar por examen médico, todos ellos laboran en el área de proceso para barras mineras. No existe una correlación significativa entre la concentración de plomo en la sangre y la edad de los trabajadores. El 74 % de los trabajadores presentan anemia leve. Existe una correlación inversa altamente significativa entre la concentración de plomo en la sangre y la concentración de hemoglobina. Conclusiones: Se determinó que la relación entre la concentración de plomo en sangre y la concentración de hemoglobina de los trabajadores de SIDERPERÚ, fue significativa. Palabras clave: Plomo, Anemia, Plumbemia, Hemoglobina, Trabajadores. / --- Introduction: Lead inhibits heme synthesis and therefore hemoglobin. Objectives: To evaluate chronic lead poisoning and its relation to problems of anemia in workers SIDERPERU. Sample population: 50 employees. Determination of blood lead: Atomic absorption spectroscopy with graphite furnace. Hemoglobin analysis: for the colorimetric determination of hemoglobin as blood hemiglobin cyanide. Results: The correlation between blood lead levels and exposure time workers Siderperu shown that there is a significant correlation between the concentration of lead in the blood and the time of exposure of workers. Regarding the correlation of blood lead levels and the workspace of workers 34 % of the study population working in the area of process mining bars, 18 % work in the construction process for iron, 16 % in the steel plant, 14 % in workshops led, 12 % in area standardized and 6 % in coils import. 6 % of workers have blood lead values that require going through medical examination, all working in the area of process mining bars. There is no significant correlation between the concentration of lead in the blood and the age of the workers. 74 % of workers have mild anemia. There is a highly significant inverse correlation between the concentration of lead in the blood and hemoglobin concentration. Conclusions: We found that the relationship between blood lead concentration and hemoglobin concentration SIDERPERU workers, was significant. Keywords: Lead, Anemia, blood lead, hemoglobin, workers. / Tesis

Aplastic anemia (AA) is a rare, life-threatening disorder characterized by immune mediated loss of hematopoietic stem cells (HSC) that results in bone marrow hypoplasia, pancytopenia and its complications. Majority of AA are acquired and idiopathic. Cytotoxic T cell-mediated autoimmune destruction to the HSC is the main pathophysiological mechanism. Intrinsic defects in the hematopoietic stem cell, alterations in bone marrow microenvironment, acquired clonal abnormalities and drug/viral direct injury are other described etiologies. Improvement in molecular studies also led to better understanding of cytogenetic abnormalities, somatic mutations, telomere attrition in AA which may affect treatment response, prognosis and survival. HSCT or immunosuppression therapy (IST) can restore normal hematopoiesis. HSCT is curative, but eligibility depends on age, comorbidities, and available donors. HSCT use is limited by matched donor availability. The alternative option is IST. The standard today is intensive triple therapy with anti-thymocyte globulin (ATG), cyclosporin (CsA) and thrombopoietin receptor agonist, eltrombopag (EPAG) or with ATG and CsA in patients who require less intense IST. 72 year-old male presented with pancytopenia (WBC 1.7K/uL, Hgb 12.9 g/dL, Platelet count 6 K/uL, and ANC 600, absolute reticulocyte count 18x10*9. Bone marrow biopsy revealed markedly hypocellular marrow ( Response rates for first line IST is about 60–70%. The response and its duration are strong surrogate markers for prolonged survival. Triple therapy with hATG, CsA and EPAG improved response rate to 90% in AA in 6 months. Bone marrow evaluation in relapse setting is essential to rule out clonal evolution into MDS/AML or subtle adult inherited bone marrow syndromes. Survival is limited after second relapse in elderly. Our patient had accelerated response and CR with sustained survival after alemtuzumab, CsA and romiplostim. This report highlights a rare response to alemtuzumab and an alternate TPO mimetic, romiplostim. Given his exposure to various methods of IST and the bone marrow showing clonal evolution, he is at risk of myeloid malignancies.

No description available.

La anemia representa en la salud pública, un grave problema a nivel mundial. El fin de este estudio fue determinar la relación entre la participación materna y la efectividad del tratamiento de la anemia ferropénica en niños de 6 a 36 meses en un centro de salud. Se realizó un estudio de tipo cuantitativo, descriptivo, de diseño correlacional y de corte transversal. Constituida por 95 participantes del C.S Paul Harris que cumplieron con los criterios de inclusión. Para la recolección de datos se realizaron encuestas que evaluaron factores sociodemográficos, características del niño, factores relacionados al cuidado materno y dosaje de hemoglobina de acuerdo al protocolo. El análisis de las variables cualitativas determinó las frecuencias absolutas y relativas. Para la medición de la asociación entre las variables se aplicó la prueba de T-Student. Dentro de los resultados del nivel de participación: 65% tuvo participación regular, 28% baja y 6% buena. Gran parte de las madres se encontraban entre las edades de 26 a 30 años con nivel educativo secundario, la mayoría de los niños participantes tenían edades entre 6 a 12 meses, de proporciones similares tanto el sexo femenino como masculino; entre otras características. Se concluye que existe relación significativa entre la participación materna y la efectividad del tratamiento. / Anemia represents public health, a serious global problem. This study had to objectives determine the relationship between maternal participation and the effectiveness of the treatment of iron deficiency anemia in children age 6 to 36 months in Paul Harris Health Center. A quantitative, descriptive, correlational, and cross-section study was carried out. It consisted of 95 participants from the C.S Paul Harris (Paul Harris Health center) who met the inclusion criteria. For data collection surveys were conducted that evaluated sociodemographic factors, characteristics of the child, factors related to maternal care, and hemoglobin dosage according to the protocol. The analysis of qualitative variables determined absolute and relative frequencies. The T-Student test was used to measure the association between the variables. Inside the results of the level of participation: 65% had regular participation, 28% had low participation and 6% had good participation. The most of the ages of the mothers were between 26 and 30 years old with high school, and most of the participating children were between 6 and 12 months of age, with similar proportions in females and males, among other characteristics. It is concluded that there is a significant relationship between maternal participation and treatment effectiveness.

Thesis (Ph.D.)--University of Adelaide, Dept. of Public Health, 2001. / Bibliography: leaves 148-163.

Thesis (Ph. D.)--Cornell University, May, 2009. / Vita. Includes bibliographical references.

Thesis (Ph. D.)--University of Virginia, 2008. / Title from title page. Includes bibliographical references. Also available online through Digital Dissertations.