HPLC determination of hemoglobins to establish reference values with the aid of statistics and informatics

The purpose of the present study was to establish reference values for hemoglobins (Hb) using HPLC, in samples containing normal Hb (AA), sickle cell trait without alpha-thalassemia (AS), sickle cell trait with alpha-thalassemia (ASH), sickle cell anemia (SS), and Hb SC disease (SC). The blood samples were analyzed by electrophoresis, HPLC and molecular procedures. The Hb A2 mean was 4.30 ± 0.44% in AS, 4.18 ± 0.42% in ASH, 3.90 ± 1.14% in SS, and 4.39 ± 0.35% in SC. They were similar, but above the normal range. Between the AS and ASH groups, only the amount of Hb S was higher in the AS group. The Hb S mean in the AS group was 38.54 ± 3.01% and in the ASH it was 36.54 ± 3.76%. In the qualitative analysis, using FastMap, distinct groups were seen: AA and SS located at opposite extremes, AS and ASH with overlapping values and intermediate distribution, SC between heterozygotes and the SS group. Hb S was confirmed by allele-specific polymerase chain reaction. The Hb values established will be available for use as a reference for the Brazilian population, drawing attention to the increased levels of Hb A2, which should be considered with caution to prevent incorrect diagnoses. ©FUNPEC-RP.

Análise das prevalências de doenças detectadas pelo programa nacional de triagem neonatal no município de Araraquara no ano de 2009

The National Neonatal Screening Program (NNSP) set up in all Brazil, aims, through planned phases of local implementation, to detect diseases such as phenylketonuria, congenital hypothyroidism, hemoglobinopathies and cystic fibrosis. The aim of this study was to assess, through a cross-sectional observational study, the prevalence of the diseases detected by the NNSP in the city of Araraquara, in records issued by the São Paulo APAE laboratory in the period between April and December 2009.The results show that Araraquara had a prevalence of phenylketonuria and congenital hypothyroidism 0.06% above the national averages of 0.01% and 0.03% respectively. With respect to hemoglobinopathies, the prevalence of sickle cell trait was 2.15% below the national average of 2.6%. The prevalence of Hb C in the city was 0.57%, similar to national values reported in the literature. Confirmed Hb Bart's had a prevalence of 0.13% in Araraquara, below the average of 0.38% for the surrounding region. The neonatal screening by heel-prick test and counseling for caregivers are important factors in reducing morbidity related to the evolution of these diseases.

Doença como experiência: as relações entre vulnerabilidade social e corpo doente enquanto fenômeno biocultural no estado do Pará

O estudo ora apresentado analisa a representação biossocial de pessoas com Anemia Falciforme (AF) no Estado do Pará, agravo entendido como um fenômeno biocultural por envolver aspectos evolutivos, genéticos, ambientais, socioeconômicos e culturais da vivência cotidiana dos indivíduos acometidos pela síndrome. A investigação aborda as sociabilidades de quarenta (40) interlocutores com AF, representando cerca de 10% dos pacientes em tratamento na Fundação Hemopa (Belém), centro de referência em doenças hematológicas do Estado, englobando a sua situação de vulnerabilidade social, suas percepções de Saúde e Doença, os tratamentos complementares (folk medicine), diagnóstico, estigmas, preconceitos, tabus e dificuldades de acesso e acessibilidade aos serviços do SUS com os quais eles convivem rotineiramente. A metodologia compreensiva e a análise de conteúdo revelam as experiências próximas dos sujeitos que diariamente convivem com as instabilidades da enfermidade. A vivência da doença, elaborada através das relações sociais, conversas, percepções e enredamentos familiares e extrafamiliares do grupo em questão, que em seu conjunto organiza sua vida social de modo sui gêneris, foram os principais dados revelados, considerando a dor física e psicológica representada pelo corpo adoecido. O habitus em relação ao estilo de vida dos sujeitos é um recorte que engloba a natureza étnico-racial da AF, ainda entendida como “doença que vem do negro” e que necessita ser desmistificada pelos profissionais de saúde que os assistem no dia-a-dia em ambulatórios de todo o Estado. Concluo sugerindo que a AF é uma doença que está atrelada aos Determinantes Sociais em Saúde, incorporando as diversas suscetibilidades dos interlocutores, que necessitam de maior sensibilidade política e dos setores de atenção básica à saúde para que as pessoas que compartilham as vicissitudes da AF possam ser incluídas socialmente.

Síntese e avaliação farmacológica de novos compostos híbridos úteis para tratamento das complicaçôes da anemia falciforme

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

O sofrimento gera luta: o impacto da anemia falciforme e da vivência do adoecimento no desenvolvimento psíquico de portadores da doença

Pós-graduação em Saúde Coletiva - FMB

Aluminum and bone: Review of new clinical circumstances associated with Al(3+) deposition in the calcified matrix of bone

International audience

Pesquisas genéticas, prognósticos morais e discriminação genética : um estudo de caso sobre traço falciforme

O debate sobre ética em pesquisa pode ser aplicado tanto ao âmbito da metodologia científica como a outras áreas do saber, como é o caso dos esportes. No campo da saúde esportiva brasileira, têm sido comuns pesquisas que fazem testes genéticos para identificar atletas com traço falciforme. Apesar da persistência de confederações esportivas brasileiras em discriminar atletas com essa característica hereditária, o traço falciforme não é uma doença. Este artigo relata o caso de uma atleta de futebol vítima de discriminação genética: identificada com o traço falciforme, ela foi considerada inapta a participar de um campeonato pela Confederação Brasileira de Futebol. O artigo analisa as repercussões da pesquisa genética para identificação do traço falciforme na ausência de cuidados éticos voltados à preservação dos direitos de quem se submete aos testes. Além disso, mostra a situação de vulnerabilidade à qual estão expostas pessoas envolvidas em pesquisas que fazem testes genéticos sem cuidados éticos ou mesmo justificativas razoáveis e cujos resultados são interpretados sob a racionalidade do determinismo biológico e do reducionismo genético. As confederações esportivas brasileiras interessadas em identificar atletas com o traço falciforme deveriam submeter esse objetivo de estudo à avaliação de Comitês de Ética em Pesquisa, pois esse é um procedimento com potencial de acarretar prejuízos aos atletas. O teste genético não pode ser considerado um ato de assistência em saúde, visto que não há doença a ser tratada. _______________________________________________________________________________ ABSTRACT

Numerical simulation of red blood cells’ motion : a review

The micro-circulation of blood plays an important role in human body by providing oxygen and nutrients to the cells and removing carbon dioxide and wastes from the cells. This process is greatly affected by the rheological properties of the Red Blood Cells (RBCs). Changes in the rheological properties of the RBCs are caused by certain human diseases such as malaria and sickle cell diseases. Therefore it is important to understand the motion and deformation mechanism of RBCs in order to diagnose and treat this kind of diseases. Although, many methods have been developed to explore the behavior of the RBCs in micro-channels, they could not explain the deformation mechanism of the RBCs properly. Recently developed Particle Methods are employed to explain the RBCs’ behavior in micro-channels more comprehensively. The main objective of this study is to critically analyze the present methods, used to model the RBC behavior in micro-channels, in order to develop a computationally efficient particle based model to describe the complete behavior of the RBCs in micro-channels accurately and comprehensively

The effect of HLA-DR genes on susceptibility to and severity of ankylosing spondylitis

Objective. To analyze the effect of HLA-DR genes on susceptibility to and severity of ankylosing spondylitis (AS). Methods. Three hundred sixty- three white British AS patients were studied; 149 were carefully assessed for a range of clinical manifestations, and disease severity was assessed using a structured questionnaire. Limited HLA class I typing and complete HLA-DR typing were performed using DNA-based methods. HLA data from 13,634 healthy white British bone marrow donors were used for comparison. Results. A significant association between DR1 and AS was found, independent of HLA-B27 (overall odds ratio [OR] 1.4, 95% confidence interval [95% CI] 1.1-1.8, P = 0.02; relative risk [RR] 2.7, 95% CI 1.5-4.8, P = 6 x 10-4 among homozygotes; RR 2.1, 95% CI 1.5-2.8, P = 5 x 10-6 among heterozygotes). A large but weakly significant association between DR8 and AS was noted, particularly among DR8 homozygotes (RR 6.8, 95% CI 1.6-29.2, P = 0.01 among homozygotes; RR 1.6, 95% CI 1.0-2.7, P = 0.07 among heterozygotes). A negative association with DR12 (OR 0.22, 95% CI 0.09-0.5, P = 0.001) was noted. HLA-DR7 was associated with younger age at onset of disease (mean age at onset 18 years for DR7-positive patients and 23 years for DR7-negative patients; Z score 3.21, P = 0.001). No other HLA class I or class H associations with disease severity or with different clinical manifestations of AS were found. Conclusion. The results of this study suggest that HLA-DR genes may have a weak effect on susceptibility to AS independent of HLA-B27, but do not support suggestions that they affect disease severity or different clinical manifestations.

Does the microbiome play a causal role in spondyloarthritis?

The purpose of this study is to review the potential causal role of the microbiome in the pathogenesis of spondyloarthritis. The method used for the study is literature review. The microbiome plays a major role in educating the immune response. The microbiome is strongly implicated in inflammatory bowel disease which has clinical and genetic overlap with spondyloarthritis. The microbiome also plays a causal role in bowel and joint disease in HLA B27/human beta 2 microglobulin transgenic rats. The mechanism(s) by which HLA B27 could influence the microbiome is unknown but theories include an immune response gene selectivity, an effect on dendritic cell function, or a mucosal immunodeficiency. Bacteria are strongly implicated in the pathogenesis of spondyloarthritis. Studies to understand how HLA B27 affects bacterial ecosystems should be encouraged.

Endoplasmic reticulum aminopeptidases in the pathogenesis of ankylosing spondylitis

There has been significant progress in our understanding of the pathogenesis of AS. The advent of genome-wide association studies has increased the known loci associated with AS to more than 40. The endoplasmic reticulum resident aminopeptidases (ERAP) 1 and 2 were identified in this manner and are of particular interest. There appears to be a genetic as well as a functional interaction of ERAP1 and 2 with HLA-B27 based on the known functions of these molecules. Recent studies on the structure, immunological effects and the peptide-trimming properties of ERAP 1 and 2 have helped to provide insight into their pathogenic potential in AS. In this review, we explore the role of ERAP 1 and 2 in the pathogenesis of AS. © The Author 2015.

Molecular background of three lethal fetal syndromes

This study identified the molecular defects underlying three lethal fetal syndromes. Lethal Congenital Contracture Syndrome 1 (LCCS1, MIM 253310) and Lethal Arthrogryposis with Anterior Horn Cell Disease (LAAHD, MIM 611890) are fetal motor neuron diseases. They affect the nerve cells that control voluntary muscle movement, and eventually result in severe atrophy of spinal cord motor neurons and fetal immobility. Both LCCS1 and LAAHD are caused by mutations in the GLE1 gene, which encodes for a multifunctional protein involved in posttranscriptional mRNA processing. LCCS2 and LCCS3, two syndromes that are clinically similar to LCCS1, are caused by defective proteins involved in the synthesis of inositol hexakisphosphate (IP6), an essential cofactor of GLE1. This suggests a common mechanism behind these fetal motor neuron diseases, and along with accumulating evidence from genetic studies of more late-onset motor neuron diseases such as Spinal muscular atrophy (SMA) and Amyotrophic lateral sclerosis (ALS), implicates mRNA processing as a common mechanism in motor neuron disease pathogenesis. We also studied gle1-/- zebrafish in order to investigate whether they would be a good model for studying the pathogenesis of LCCS1 and LAAHD. Mutant zebrafish exhibit cell death in their central nervous system at two days post fertilization, and the distribution of mRNA within the cells of mutant zebrafish differs from controls, encouraging further studies. The third lethal fetal syndrome is described in this study for the first time. Cocoon syndrome (MIM 613630) was discovered in a Finnish family with two affected individuals. Its hallmarks are the encasement of the limbs under the skin, and severe craniofacial abnormalities, including the lack of skull bones. We showed that Cocoon syndrome is caused by a mutation in the gene encoding the conserved helix-loop-helix ubiquitous kinase CHUK, also known as IκB kinase α (IKKα). The mutation results in the complete lack of CHUK protein expression. CHUK is a subunit of the IκB kinase enzyme that inhibits NF-κB transcription factors, but in addition, it has an essential, independent role in controlling keratinocyte differentiation, as well as informing morphogenetic events such as limb and skeletal patterning. CHUK also acts as a tumor suppressor, and is frequently inactivated in cancer. This study has brought significant new information about the molecular background of these three lethal fetal syndromes, as well as provided knowledge about the prerequisites of normal human development.

Achados fundoscópicos em crianças portadoras de anemia falciforme no estado do Rio Grande do Norte

Detectar os principais achados fundoscópicos em crianças portadoras de hemoglobinopatias falciformes. Métodos: Foram estudados 26 pacientes com hemoglobinopatias falciformes, no Serviço de Oftalmologia do Hospital Universitário Onofre Lopes, Natal, RN, que foram submetidos a protocolo de pesquisa pré-estabelecido. Os resultados foram avaliados estatisticamente pelo teste qui-quadrado. Resultados: A idade média foi de 10,6 anos, com acuidade visual igual ou melhor que 20/25 na maioria, excetuando-se 3 olhos, que apresentavam outras doenças associadas. O tipo mais freqüente foi o SS com 57,7% (15/26) dos casos, seguido pelos SC e SA com 15,4% (4/26) cada, e pelo S-Thal com 11,5% (3/26). A freqüência da retinopatia por células falciformes foi maior após os 10 anos de idade, sendo mais freqüente, em valores relativos, no tipo S-Thal (100% dos casos) e, em valores absolutos, no tipo SS (9 casos). Os dois achados mais comuns foram tortuosidade venosa (12/26) e “black sunburst” (7/26). Conclusões: Observamos que a incidência de retinopatia por células falciformes aumentou após os 10 anos de idade e não evidenciamos achados da doença proliferativa. Portanto, enfatizamos a necessidade do exame oftalmológico precoce nos portadores de anemia falciforme, como forma de prevenir futuras complicações oculares

Anestesia em paciente obstétrica portadora de anemia falciforme e traço talassêmico após plasmaféresis: relato de caso

JUSTIFICATIVA E OBJETIVOS: A plasmaféresis é a técnica de tratamento de escolha para pacientes com anemia hemolítica grave. Uma de suas conseqüências é a depleção de colinesterase plasmática, o que interfere na metabolização de alguns bloqueadores neuromusculares de uso corrente na prática anestesiológica. RELATO do CASO: Paciente com 26 anos, estado físico ASA IV, gestação de 30 semanas e 3 dias, portadora de anemia falciforme, traço talassêmico e alo-imunização para antígenos de alta freqüência. Apresentou crise de falcização, sendo transfundida com derivado sangüíneo incompatível. Evoluiu com hemólise maciça, sendo admitida com hemoglobina de 3 g/dL e hematócrito de 10%, icterícia intensa, taquicardia, apatia e descoramento. Na avaliação hematológica concluiu-se ser situação de inexistência de sangue compatível para transfusão. Foi tratada com corticoterapia, imunoglobulinas e plasmaféresis. No segundo dia de internação, evoluiu com insuficiência renal aguda e edema pulmonar agudo, piora do estado geral e instabilidade hemodinâmica. Indicada a resolução da gestação em decorrência do quadro clínico da paciente e do sofrimento fetal agudo que se sobrepôs. A paciente foi admitida na sala de operações consciente, dispnéica, pálida, ictérica, SpO2 de 91% em ar ambiente, freqüência cardíaca de 110 bpm e pressão arterial de 110 x 70 mmHg, em uso de dopamina (1 µg.kg-1.min-1) e dobutamina (10 µg.kg-1.min-1). Optou-se por anestesia geral balanceada, com alfentanil (2,5 mg), etomidato (14 mg) e atracúrio (35 mg) e isoflurano. Não se observou intercorrências anestésico-cirúrgicas. Ao final, a paciente foi encaminhada à UTI, sob intubação orotraqueal, e em uso de drogas vasoativas, tendo sido extubada após 3 horas. CONCLUSÕES: Este caso mostrou-se um desafio para a equipe, visto que a paciente apresentava instabilidade hemodinâmica e alteração do coagulograma, condições que contra-indicam a anestesia regional; além disto, a plasmaféresis potencialmente depleta os estoques de colinesterases plasmáticas, o que interfere na anestesia. Entretanto, o arsenal medicamentoso disponível permitiu o manuseio seguro desta situação.

Determinação de haplótipos do gene βs em portadores de anemia falcifome

Haplotypes linked to the βS gene represent patterns of DNA polymorphisms along chromosome 11 of individuals bearing the βS gene. Analysis of haplotypes, in addition to serving as an important source for anthropological studies about the ethnic origin of a population, contributes to a better understanding of the variations in clinical severity of sickle cell anemia. The aim of the present study was to determine βS gene haplotypes in a group of patients with sickle cell anemia treated at the Dalton Barbosa Cunha Hematology Center (Hemonorte) in Natal, Brazil and the Oncology and Hematology Center in Mossoró, Brazil. Blood samples were obtained from 53 non-related patients (27 males and 26 females), aged between 3 months and 61 years (mean age: 16.9 ± 12.1 years). Laboratory analyses consisted of the following: erythrogram, reticulocyte count, hemoglobin electrophoresis at alkaline pH, measurement of hemoglobin A2 and Fetal hemoglobin, solubility test and molecular analysis to determine βS gene haplotypes. DNA samples were extracted by illustra blood genomicPrep Mini Spin kit and βS gene haplotypes were determined by PCR-RFLP, using Xmn I, Hind III, Hinc II and Hinf I restriction enzymes for analysis of six polymorphic restriction sites in the beta cluster. Of 106 βS chromosomes studied, 75.5% were Central African Republic (CAR) haplotype, 11.3% Benin (BEN) and 6.6% Cameroon (CAM). The atypical haplotypes had a frequency of 6.6%. More than half the patients (58.5%) were identified as CAR/CAR genotype carriers, 16.9% heterozygous CAR/BEN, 13.2% CAR/CAM and 1.9% BEN/BEN. Patients with atypical haplotype in one or two chromosomes accounted for 9.5% (CAR/Atp, BEN/Atp and Atp/Atp). The genotype groups showed no statistically significant difference (p < 0.05) in their laboratory parameters. This is the first study related to βS haplotypes conducted in state of Rio Grande do Norte and the higher frequency of Cameroon halotype found, compared to other Brazilian states, suggests the existence of a peculiarity of African origin