La drépanocytose est une maladie génétique touchant l'hémoglobine, de transmission autosomique récessive, caractérisée par trois grandes manifestations cliniques :anémie hémolytique chronique, phénomènes vaso-occlusifs et susceptibilité accrue aux infections. Dans diverses régions du monde et particulièrement en Afrique subsaharienne, cette maladie est très fréquente et constitue un problème crucial de santé publique. Sa physiopathologie complexe est basée sur la polymérisation de l’hémoglobine anormale (Hb S) qui provoque une falciformation et une déshydratation du globule rouge. Les hématies falciformées sont impliquées dans les phénomènes de vaso-occlusion. Le traitement et la prise en charge de la maladie reste toujours problématique. A l’heure actuelle, le seul traitement curatif est la transplantation de moelle osseuse mais les donneurs compatibles sont assez rares et les coûts élevés. Des traitements symptomatiques et préventifs (principalement la transfusion et l’hydroxyurée) existent toutefois.<p>Des études in vitro et in vivo ont démontré les possibilités thérapeutiques de certaines molécules dont les cibles sont les transports membranaires impliqués dans la déshydratation cellulaire.<p>Depuis les années 1990, le cromoglycate de sodium, un médicament anti-allergique et anti-asthmatique, a montré un intérêt potentiel dans le traitement de la drépanocytose. Néanmoins, son mode d’action n’est actuellement pas connu. Notre travail a pour but de contribuer à l’étude du mécanisme d’action anti-drépanocytaire de la molécule.<p>Dans un premier temps, des globules rouges drépanocytaires préincubés en absence ou présence de cromoglycate ont été désoxygénés par un flux d’azote. Ensuite, les concentrations intracellulaires en Na+ et en K+ ont été mesurées. Les résultats de ces investigations ont montré un effet inhibiteur du cromoglycate sur l’efflux de K+ et l’influx du Na+ provoqués par la désoxygénation.<p>Sur base de ces observations, des expériences testant l’action du cromoglycate sur le canal K+ dépendant du Ca2+ (canal de Gardos) ont été effectuées. Dans des globules rouges normaux et drépanocytaires, ce canal a été activé par augmentation de la concentration intra-cellulaire en Ca2+. L'effet du cromoglycate a été comparé à celui d'un inhibiteur connu, le clotrimazole. Les résultats ont montré que 1e cromoglycate n'exerce pas d'effet inhibiteur sur le canal de Gardos, au contraire du clotrimazole. Il est également sans effet significatif sur la Ca2+-ATPase.<p>Nous avons ensuite investigué l’implication du Ca2+ dans les perturbations du flux des ions K+ et Na+. Des globules rouges drépanocytaires ont été incubés en absence et présence d’EGTA 5 mmol/l ou de BAPTA 10 µmol/l, respectivement chélateurs du Ca2+ extra et intracellulaire. Après désoxygénation, les concentrations intracellulaires en Na+ et K+ ont été mesurées. Les résultats de ces expériences montrent que seul le chélateur du Ca2+ extracellulaire bloque les perturbations ioniques causées par la désoxygénation. Ces résultats viennent donc confirmer les observations d’autres auteurs quant à l’implication du Ca2+ extracellulaire dans la fuite de K+ des globules drépanocytaires soumis à la désoxygénation. <p>Enfin, l’effet du cromoglycate sur l’influx de Ca2+ extracellulaire et sur la falciformation induits par le métabisulfite a été mesuré et comparé à celui du clotrimazole. Des globules rouges drépanocytaires, prélablement chargés en Fura Red, un indicateur fluorescent du Ca2+, ont été exposés au métabisulfite, un puissant réducteur provoquant une falciformation rapide. L’influx de Ca2+ a été mesuré par la diminution de la fluorescence du Fura Red. Parallèlement, la falciformation a été suivie en mesurant la lumière diffractée à 90° par les cellules. Les résultats de ces investigations montrent que le cromoglycate (1 µmol/l) et le clotrimazole (10 µmol/l) ont des effets inhibiteurs comparables sur la falciformation mais que le cromoglycate freine significativement plus l'influx de Ca2+ au cours de ce processus.<p>En conclusion, sur base de ces différents tests in vitro, le cromoglycate inhibe la falciformation induite par la désoxygénation. Cette inhibition résulte du blocage des perturbations ioniques induites par la désoxygénation en empêchant l’influx du Ca2+ extracellulaire et secondairement la fuite du K+ intracellulaire, ce qui inhibe la déshydratation cellulaire.<p>La diminution des crises vaso-occlusives observée chez les patients drépanocytaires traités par le cromoglycate s’expliquerait donc par ces effets. En présence de cromoglycate, les globules rouges sont moins déshydratés et falciforment moins rapidement. Ils sont dès lors moins impliqués dans les phénomènes de vaso-occlusion, ce qui améliore l’état des patients.<p> / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished

Indiana University-Purdue University Indianapolis (IUPUI) / High-efficiency transduction of hematopoietic stem cells (HSCs) by recombinant adeno-associated virus serotype 2 (AAV2) vectors is limited by (i) inadequate expression of cellular receptor/co-receptors for AAV2; (ii) impaired intracellular trafficking and uncoating in the nucleus; (iii) failure of the genome to undergo second-strand DNA synthesis; and (iv) use of sub-optimal promoters. Systematic studies were undertaken to develop alternative strategies to achieve high-efficiency transduction of primary murine HSCs and lineage-restricted transgene expression in a bone marrow transplant model in vivo. These included the use of: (i) additional AAV serotype (AAV1, AAV7, AAV8, AAV10) vectors; (ii) self-complementary AAV (scAAV) vectors; and (iii) erythroid cell-specific promoters. scAAV1 and scAAV7 vectors containing an enhanced green-fluorescent protein (EGFP) reporter gene under the control of hematopoietic cell-specific enhancers/promoters allowed sustained transgene expression in an erythroid lineage-restricted manner in both primary and secondary transplant recipient mice. Self complementary AAV vectors containing an anti-sickling human beta-globin gene under the control of either the beta-globin gene promoter/enhancer, or the human parvovirus B19 promoter at map-unit 6 (B19p6) were tested for their efficacy in a human erythroid cell line (K562), and in primary murine hematopoietic progenitor cells (c-kit+, lin-). These studies revealed that (i) scAAV2-beta-globin vectors containing only the HS2 enhancer are more efficient than ssAAV2-beta-globin vectors containing the HS2+HS3+HS4 enhancers; (ii) scAAV-beta-globin vectors containing only the B19p6 promoter are more efficient than their counterparts containing the HS2 enhancer/beta-globin promoter; and (iii) scAAV2-B19p6-beta-globin vectors in K562 cells, and scAAV1-B19p6-beta-globin vectors in murine c-kit+, lin- cells, yield efficient expression of the beta-globin protein. These studies suggest that the combined use of scAAV serotype vectors and the B19p6 promoter may lead to expression of therapeutic levels of beta-globin gene in human erythroid cells, which has implications in the potential gene therapy of beta-thalassemia and sickle cell disease.

No description available.

Symmetry in biological systems is the occurrence of an event on both sides of the system. The term bilateralism was introduced to represent this phenomenon, and it was defined as the conditional co-occurrence of two events given that at least at one of them has occurred. This phenomenon is highly associated with the prevalence of each of the events. Two parameters were developed to evaluate the presence of this phenomenon, testing whether events co-occur with higher probability than would be expected by chance. Nonparametric confidence intervals were constructed using the bootstrap percentile method. These non parametric confidence intervals were used in testing the null hypothesis of no bilateralism.A simulation study was performed to examine the properties of the two bilateralism parameters' estimates. The size and power of the tests of bilateralism were examined under a variety of sample sizes and prevalences of the two events. The simulation study showed that both parameter estimates have similar properties, and the tests have similar size and power. The power of the test was affected by the prevalence of either event, the differences in the prevalences, the sample size and by number of events that occur simultaneously. The methodology of testing for bilateralism was applied on data from the Pain in Sickle Cell Epidemiology Study (PiSCES). This study collected up to 6 months worth of daily diaries about pain and medical utilization from patients with sickle cell disease. Each diary recorded the body site and side where pain was experienced over the past 24 hours. The sample consists of 119 subjects who completed at least 50 daily pain diaries (reference). Information about the subjects age, gender and sickle cell genotype were also available. Nine body sites (5 upper peripheral, and 4 lower peripheral site) were analyzed to test for bilateralism. Bilateralism was tested for each subject and each site separately. The associations of prevalence of bilateralism on each site, and percentages of sites that hurt bilaterally with age, gender and genotype where studied.The results show a high prevalence of bilateral pain among sickle cell patients at all sites. Age gender and genotype were associated with higher prevalence in bilateral pain in some, but not all sites. The percentage of sites that have bilateral pain is also associated with the number of sites that have pain.

Introduction. 5-HMF and novel INN-compounds are left-shifting AEH, shown to have anti-sickling action by forming transiently covalent Schiff-base adducts with hemoglobin (Hb), thereby increasing the Hb O2-affinity. They are hypothesized to be substrates for aldehyde dehydrogenase (ALDH) in the liver and red blood cells (RBC). Methods. Biopharmaceutical assessments were made for AEH, using calculated physicochemical properties. Their in-vitro hepatic metabolism (mediated by ALDH) was characterized using hepatic cytosol, and in-vitro-in-vivo extrapolations (IVIVE) were made. Inter-species differences in hepatic cytosolic ALDH activity were investigated using acetaldehyde as a model substrate in different mammalian species. Time- and concentration-dependent in-vitro disposition of 5-HMF in human whole blood was fully characterized and quantitatively modeled. In-vitro time- and concentration-dependent pharmacodynamic (PD) profiling of AEH (0.5 – 5 mM) was carried out in normal whole blood. 5-HMF binding to (normal) HbA and (sickle) HbS was studied in systematic time- and concentration-dependency studies using isolated Hb solutions. Quantitative PK/PD models were developed to fit the experimental data by nonlinear regression (Scientist®). Results. 5-HMF and the two INN-compounds were classified as BCS-I and BCS-II, respectively. All AEH were substrates for hepatic ALDH, with predicted low/intermediate hepatic extraction. Intrinsic ALDH activity varied significantly between mammalian species. In whole blood, 5-HMF plasma concentrations declined rapidly (t1/2 of 0.8 – 4 hrs), with nonlinear kinetics, due to saturable Hb-binding. AEH showed a time-dependent, biphasic PD effect in whole blood, suggesting transiently covalent Hb binding, with slow recovery to the baseline, corresponding to dissociation from Hb and subsequent metabolism by RBC-ALDH. Binding studies with HbA and HbS demonstrated slight differences in binding affinity, but sustained adduct formation - with slow dissociation t1/2. A novel semi-mechanistic target-site drug disposition (TSDD)/PD model was developed, integrating the information, for simultaneous modeling of 5-HMF concentrations in plasma, and its effect in whole blood. Conclusions. This translational research investigated in detail the in-vitro PK/PD of AEH, and systematically compared findings with older generation compounds. A (generic) novel TSDD/PD model was developed for disposition of AEH, identifying k-1 (dissociation constant of AEH from Hb) and kmet (RBC-ALDH metabolism rate constant) as key properties for the time course of PD effect.

Red blood cell hemolysis in sickle cell disease (SCD) releases free hemoglobin. Extracellular hemoglobin and its degradation products, free heme and iron, are highly toxic due to oxidative stress induction and decrease in nitric oxide availability. We propose an approach that helps to eliminate extracellular hemoglobin toxicity in SCD by employing a bacterial protein system that evolved to extract heme from extracellular hemoglobin. NEAr heme Transporter (NEAT) domains from iron-regulated surface determinant proteins from Staphylococcus aureus specifically bind free heme as well as facilitate its extraction from hemoglobin. We demonstrate that a purified NEAT domain fused with human haptoglobin beta-chain is able to remove heme from hemoglobin and reduce heme content and peroxidase activity of hemoglobin. We further use molecular dynamics (MD) simulations to resolve molecular pathway of heme transfer from hemoglobin to NEAT, and to elucidate molecular mechanism of such heme transferring process. Our study is the first of its kind, in which simulations are employed to characterize the process of heme leaving hemoglobin and subsequent rebinding with a NEAT domain. Our MD results highlight important amino acid residues that facilitate heme transfer and will guide further studies for the selection of best NEAT candidate to attenuate free hemoglobin toxicity.

<p>Background: Sickle Cell Disease (SCD) is a genetic hematological disorder that affects more than 7 million people globally (NHLBI, 2009). It is estimated that 50% of adults with SCD experience pain on most days, with 1/3 experiencing chronic pain daily (Smith et al., 2008). Persons with SCD also experience higher levels of pain catastrophizing (feelings of helplessness, pain rumination and magnification) than other chronic pain conditions, which is associated with increases in pain intensity, pain behavior, analgesic consumption, frequency and duration of hospital visits, and with reduced daily activities (Sullivan, Bishop, & Pivik, 1995; Keefe et al., 2000; Gil et al., 1992 & 1993). Therefore effective interventions are needed that can successfully be used manage pain and pain-related outcomes (e.g., pain catastrophizing) in persons with SCD. A review of the literature demonstrated limited information regarding the feasibility and efficacy of non-pharmacological approaches for pain in persons with SCD, finding an average effect size of .33 on pain reduction across measurable non-pharmacological studies. Second, a prospective study on persons with SCD that received care for a vaso-occlusive crisis (VOC; N = 95) found: (1) high levels of patient reported depression (29%) and anxiety (34%), and (2) that unemployment was significantly associated with increased frequency of acute care encounters and hospital admissions per person. Research suggests that one promising category of non-pharmacological interventions for managing both physical and affective components of pain are Mindfulness-based Interventions (MBIs; Thompson et al., 2010; Cox et al., 2013). The primary goal of this dissertation was thus to develop and test the feasibility, acceptability, and efficacy of a telephonic MBI for pain catastrophizing in persons with SCD and chronic pain. </p><p>Methods: First, a telephonic MBI was developed through an informal process that involved iterative feedback from patients, clinical experts in SCD and pain management, social workers, psychologists, and mindfulness clinicians. Through this process, relevant topics and skills were selected to adapt in each MBI session. Second, a pilot randomized controlled trial was conducted to test the feasibility, acceptability, and efficacy of the telephonic MBI for pain catastrophizing in persons with SCD and chronic pain. Acceptability and feasibility were determined by assessment of recruitment, attrition, dropout, and refusal rates (including refusal reasons), along with semi-structured interviews with nine randomly selected patients at the end of study. Participants completed assessments at baseline, Week 1, 3, and 6 to assess efficacy of the intervention on decreasing pain catastrophizing and other pain-related outcomes. </p><p>Results: A telephonic MBI is feasible and acceptable for persons with SCD and chronic pain. Seventy-eight patients with SCD and chronic pain were approached, and 76% (N = 60) were enrolled and randomized. The MBI attendance rate, approximately 57% of participants completing at least four mindfulness sessions, was deemed acceptable, and participants that received the telephonic MBI described it as acceptable, easy to access, and consume in post-intervention interviews. The amount of missing data was undesirable (MBI condition, 40%; control condition, 25%), but fell within the range of expected missing outcome data for a RCT with multiple follow-up assessments. Efficacy of the MBI on pain catastrophizing could not be determined due to small sample size and degree of missing data, but trajectory analyses conducted for the MBI condition only trended in the right direction and pain catastrophizing approached statistically significance. </p><p>Conclusion: Overall results showed that at telephonic group-based MBI is acceptable and feasible for persons with SCD and chronic pain. Though the study was not able to determine treatment efficacy nor powered to detect a statistically significant difference between conditions, participants (1) described the intervention as acceptable, and (2) the observed effect sizes for the MBI condition demonstrated large effects of the MBI on pain catastrophizing, mental health, and physical health. Replication of this MBI study with a larger sample size, active control group, and additional assessments at the end of each week (e.g., Week 1 through Week 6) is needed to determine treatment efficacy. Many lessons were learned that will guide the development of future studies including which MBI strategies were most helpful, methods to encourage continued participation, and how to improve data capture.</p> / Dissertation

A anemia falciforme (AF) é uma doença monogênica de elevada mortalidade e morbidade, que afeta milhões de pessoas em todo o mundo. Não há tratamento definitivo que seja amplo, eficaz e seguro para a AF, de forma que os tratamentos paliativos são os mais utilizados. O tratamento definitivo disponível é transplante alogênico de células-tronco hematopoiéticas, porém com várias complicações envolvidas. O estabelecimento de um modelo in vitro permite uma melhor compreensão de como a doença ocorre, além de permitir o desenvolvimento de novos testes e tratamentos mais eficazes contra a doença. Neste contexto, a tecnologia das células-tronco pluripotentes induzidas (iPSC), que surgiu em 2006, é uma ferramenta poderosa na pesquisa básica, na pesquisa da diferenciação de tecidos e no modelamento de doenças, e uma promessa para futuras aplicações clínicas, na descoberta e triagem de novas drogas mais eficazes e seguras, além da possibilidade de utilização na medicina regenerativa, na produção de células paciente-específicas para terapia celular. Este trabalho teve como objetivo obter um modelo de estudo e tratamento da AF utilizando iPSC. Para isso, vetores epissomais foram utilizados para a reprogramação de células mononucleares de sangue periférico para obter iPSC livres de integração. Estas células foram coletadas de pacientes tratados com o medicamento hidroxiureia e sem tratamento, para avaliação do impacto da droga na reprogramação. As linhagens de iPSC PBscd geradas foram caracterizadas quanto ao potencial pluripotente e de diferenciação. Todas as linhagens geradas se mostraram pluripotentes com potencial de auto renovação e potencial de formar células e tecidos dos 3 folhetos germinativos. O rastreamento dos vetores utilizados na reprogramação mostrou que as células estão livres após cerca de 10 passagens em média, e que eles não se integram espontaneamente nas células. As linhagens de iPSC foram diferenciadas em progenitores hematopoiéticos através da agregação forçada associada à indução com citocinas específicas e um cultivo em suspensão. Dessa forma, nós obtivemos um protocolo dinâmico e eficiente de produção de células CD34+CD45+ com poucos dias de indução. Foram realizados experimentos iniciais de padronização da metodologia de CRISPR, para que essa metodologia possa ser utilizada no futuro para a correção da mutação da AF no gene da ?- globin. Além disso, a reação padronizada para o rastreamento da mutação no gene da ?-globin poderá ser usado em experimentos futuros de edição gênica para avaliar a correção da mutação. Em resumo, oferecemos uma ferramenta valiosa para uma melhor compreensão de como a AF ocorre, além de tornar possível o desenvolvimento de drogas e tratamentos mais eficazes e de fornecer um melhor entendimento dos tratamentos amplamente utilizados, como a hidroxiurea / Sickle cell anemia (SCA) is a monogenic disease of high mortality and morbidity, that affects millions of people worldwide. There is no definitive treatment that is broad, effective and safe for SCA, so the palliative treatments are the most used. The definitive treatment available is the allogeneic transplantation of hematopoietic stem cells, but with several complications involved. The establishment of an in vitro model allows better understanding of how the disease occurs, besides allowing the development of more effective new tests and treatments against the disease. In this context, the induced pluripotent stem cell (iPSC) technology, that emerged in 2006, is a powerful tool for basic research, tissue differentiation research and disease modeling, and a promise for future clinical applications, to find and screen new, more effective and safe drugs, besides the possibility of use in regenerative medicine, in the production of patient-specific cells for cell therapy. This work aimed to obtain a model for study and treatment of SCA using iPSC. For this, episomal vectors were used for reprogramming peripheral blood mononuclear cells to obtain integration-free iPSC. This cells were collected from patients treated with hydroxyurea and without treatment, for evaluation of the impact of the drug in reprogramming. The generated iPSC PBscd lines were characterized for pluripotent and differentiation potential. All the generated lines were shown to be pluripotent with potential for self-renewal and to form cells and tissues of the 3 germ layers. Screening of the vectors used for reprogramming showed that they are absent after about 10 passages, and that they do not integrate spontaneously into the cells. The iPSC lines were differentiated into hematopoietic progenitors through forced aggregation associated with induction with specific cytokines and culture in cell suspension. Thus, we obtained a dynamic and efficient protocol of CD34+CD45+ cells production with a few days of induction. Initial standardization experiments of CRISPR methodology was performed, so that this methodology can be used in the future to correct the ?-globin chain mutation of SCA. Also, the standardized reaction for the screening of ?-globin chain mutation can be used in future gene-editing experiments to evaluate the mutation correction. In summary, we offer a valuable tool for a better understanding of how SCA occurs, in addition to make possible the development of more effective drugs and treatments and providing better understanding of widely used treatments, such hydroxyrea

Orientador: Sonia Marta Moriguchi / Resumo: Introdução: A doença falciforme (DF) é a doença monogênica herdada mais comum no mundo. Acometimentos cardiopulmonares de caráter progressivo associados à vaso oclusão e fenômenos embólicos assim como, as exacerbações de sintomas respiratórios são causas recorrentes de internações nessa população. A cintilografia de perfusão é método de imagem sensível e consagrado para avaliação hipoperfusão pulmonar, notadamente em processos embólicos. A semi-quantiticação da porcentagem de obstrução vascular de perfusão (POVF) apresenta relação segura com a angiografia. Objetivo: 1) Verificar a correlação entre os sintomas clínicos (Classes funcionais) versus Porcentagem de defeitos perfusionais (POVF%) em portadores de DF e 2) Verificar a associação entre POVF% e espirometria, avaliada pela porcentagem da capacidade vital forçada do predito (CVF%). Casuística e Metodologia Trata-se de estudo transversal observacional descritivo de 22 portadores de Doença Falciforme (DF), com coleta retrospectiva de dados clínicos e de espirometria, no período de 2017-2019 e semi-quantificação da cintilografia de perfusão pulmonar (POVF%), CF de I a IV e CVF% Análise estatística descritiva e inferenciais (Coeficiente de correlação – CC), com nível de significância com p<0,05. Resultados: Foram observados cinco subtipos de DF: SS (n=13), BSB0 (n=04), SS+alfatalassemia (n=02), SS+ fetal elevada (n=02) e SC (n=1). A cintilografia de perfusão pulmonar foi sensível identificando defeitos perfusionais em 91% da ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: INTRODUCITON: Sickle cell disease (SCD) is the most common inherited monogenic disease in the world. Cardiopulmonary complications of a progressive character, associated with vessel occlusion and embolic phenomena, such as, exacerbations of respiratory symptoms are recurrent causes of hospitalization in this population. Perfusion scintigraphy is a sensitive and conspicuous imaging method for evaluating pulmonary hypoperfusion, especially in embolic processes. The semi-quantification of the percentage of vascular perfusion obstruction (POVF) presents a safe relationship with angiography. Objective: 1) To verify the correlation between clinical symptoms (functional classes- FC) versus percentage of perfusion defects (POVF%) in patients with CSD and 2) to verify the association between POVF% and spirometry, evaluated by the percentage of forced vital capacity predicted VCF%). MATERIALS AND METHODOLOGY This was a descriptive, observational cross-sectional study of 22 patients with SCD, with retrospective collection of clinical data and spirometry data for the period 2017-2019 and semi-quantification of pulmonary perfusion scintigraphy (POVF%) , FC from I to IV and VCF% Descriptive and inferential statistical analysis (Coefficient of correlation - CC), with level of significance with p <0.05. Results: Five subtypes of SCD: SS (n = 13), BSB0 (n = 04), SS + alphatassemia (n = 02), high fetal SS + (n = 02) and SC (n = 1) were observed. Pulmonary perfusion scintigraphy was sensitive b... (Complete abstract click electronic access below) / Mestre

Segundo estimativas da Organização Mundial de Saúde (OMS) no Brasil nascem 3.500 crianças com anemia falciforme a cada ano e 20% delas não conseguem atingir os cinco anos de idade devido às complicações diretas causadas pela doença. A anemia falciforme é uma doença do grupo das hemoglobinopatias extremamente comum e é causada por uma alteração molecular no cromossoma 11, quando ocorre a substituição do ácido glutâmico pelo aminoácido valina, na posição 6 da cadeia da &beta;- globina. No Brasil acredita-se que a anemia falciforme seja a doença hereditária que mais prevalece, sendo assim, ela é considerada um problema de saúde pública. Alguns estudos apontaram a existência de uma prevalência de aproximadamente 4 a 5% em recém-nascidos portadores da doença e sua maior incidência ocorre no norte e nordeste, regiões brasileiras mais pobres e com maior miscigenação das populações. É urgente a necessidade do desenvolvimento de um diagnóstico viável e acessível a tais áreas, pois existe o reconhecimento pela própria OMS de tal prioridade e que levou o Brasil a criar uma Portaria em 2005 e um Decreto em 2008 que instituem atenção integral aos portadores da doença falciforme. Portanto, foi desenvolvido neste trabalho um método de triagem para a detecção de anemia falciforme, que usou uma tecnologia atual conhecida como point-of-care. Para isso foi utilizado uma gota de sangue em meio tamponado com detergente comercial Limpol® para lisar as hemácias e o hidrossulfito de sódio para reduzir a hemoglobina S e esta foi detectada em papel de cromatografia. O método foi desenvolvido com eficácia, pois foi proposta a substituição da saponina (reagente caro) por detergente comercial Limpol® e do hidrossulfito de sódio (reagente caro) por tiossulfato de sódio, tornando o custo do kit desenvolvido muito baixo. Portanto, sob quaisquer condições adversas e em qualquer região que necessite da triagem diagnóstica populacional, principalmente nas comunidades carentes e de difícil acesso, será possível levar uma solução simples a campo para a execução de um diagnóstico rápido e sem aumento do ônus para o sistema de saúde público brasileiro, consolidando os direitos adquiridos dos indivíduos, mas que não abrangem àqueles que mais necessitam. / According to estimates from the World Health Organization (WHO), every year are born around 3.500 children with sickle cell anemia (SCA) in Brazil and 20% of them wont reach five years old because of complications directly related to the disease. The SCA is a disease of the hemoglobinopathies group extremely common and is caused by a molecular alteration in the chromosome 11, when occurs the replacement from the acid glutamic acid by amino acid valine in the chain of position 6 of the &beta;- globin. In Brazil is believed that the SCA is the hereditary disease that prevails more being considered as a public health problem. Some studies have indicated that exist prevalence in approximately 4-5% of this disease in newborns and the greatest incidence there is mainly in north and northeast, regions with bigger miscegenation from population. Is urgent the necessity in to develop viable accessible diagnostic for those areas whereas it was recognized as a priority by the WHO leading Brazil to create an Ordinance in 2005 and a Decree in 2008 establishing total care to the people with sickle cell disease. Therefore, was developed in this work a screening method to detect the sickle cell anemia, which used a technology known as point-of-care. Was created an alternative method to substitute one international kit which has high cost for the adequation according with of economic Public Health conditions from Brazil. For this a drop of blood in buffered medium with liquid detergent Limpol®, to lyse RBC and the (tiossulfato de sódio) to reduce the RBC S which was detected in simple celulose paper. The method was developed with efficiency, on this account the substitution of saponina (expensive reagent) by liquid detergent Limpol® and (hidrossulfitob de sodio) (expensive reagent) by (tiossulfato de sódio) was successful making the cost of developed kit cheaper. Therefore in any population with adverse conditions or region where there are needs, mainly in poor communities with difficult acces, will be possible to lead a simple solution at field to execution by a quickly diagnostic without increasing of charges to health public brazilian system, consolidating acquired rights to individuals that not are reached by those needs yet.