Background: Brazil has a multiethnic population with a high diversity of hemoglobinopathies. While screenings for beta-globin mutations are far more common, alterations affecting alpha-globin genes are usually more silent and less well known. The aim of this study was to describe the results of a screening program for alpha-globin gene mutations in a representative sample of the Southeastern Brazilian population. Methods: A total of 135,000 individuals, including patients with clinical suspicion of hemoglobinopathies and their family members, randomly chosen individuals submitted to blood tests and blood donors who were abnormal hemoglobin carriers were analyzed. The variants were screened by alkaline and acid electrophoreses, isoelectric focusing and cation-exchange high performance liquid chromatography (HPLC) and the abnormal chains were investigated by reverse-phase high performance liquid chromatography (RP-HPLC). Mutations were identified by molecular analyses, and the oxygen affinity, heme-heme cooperativity and Bohr effect of the variants were evaluated by functional tests. Results: Four new and 22 rare variants were detected in 98 families. Some of these variants were found in co-inheritance with other hemoglobinopathies. Of the rare hemoglobins, Hasharon, Stanleyville II and J-Rovigo were the most common, the first two being S-like and associated with alpha-thalassemia. Conclusion: The variability of alpha-globin alterations reflects the high degree of racial miscegenation and an intense internal migratory flow between different Brazilian regions. This diversity highlights the importance of programs for diagnosing hemoglobinopathies and preventing combinations that may lead to important clinical manifestations in multiethnic populations.

Hemoglobinopathies are the most common recessive diseases worldwide but their prevalence in Uruguay has not been investigated. In this study, 397 unrelated outpatient children from the Pereira Rosell Hospital Center (CHPR), as well as 31 selected patients with microcytic anemia and 28 β-thalassemia carriers were analyzed for hemoglobinopathies by using biochemical and molecular biology methods. Parametric and non-parametric methods were used to compare the hematological indices between groups of genotypes. Of the 397 patients in the first group, approximately 1% (0.76% HbS and 0.25% β-thalassemia) had a mutation in the HBB gene and 3.3% had α-thalassemia. These mutations had a heterogeneous distribution that varied according to individual ancestry. HbS was found exclusively in individuals with declared African ancestry and had a carrier frequency of 2.2%. The frequency of α-thalassemia carriers in outpatients of European and African ancestry was 1.2% and 6.5%, respectively. In contrast, the frequency of α-thalassemia carriers in patients with microcytic anemia was 25.8%, significantly higher (p < 0.01) than that observed in the sample as a whole and in Afro-descendants and Euro-descendants. Significant differences were observed in the hematological parameters between individuals with thalassemia genotypes and those with a normal genotype. These results indicate that hemoglobinopathies are a relevant health problem in Uruguay.

α-Thalassemia, arising from a defect in a-globin chain synthesis, is often caused by deletions involving one or both of the a-genes on the same allele. With the aim of investigating the prevalence of α-thalassemia 3.7 kb deletion in the adult population of Rio Grande do Norte, 713 unrelated individuals, between 18 and 59 years-of-age, were analyzed. Red blood cell indices were electronically determined, and A2 and F hemoglobins evaluated by HPLC. PCR was applied to the molecular investigation of α-thalassemia 3.7 kb deletion. Eighty (11.2%) of the 713 individuals investigated presented α-thalassemia, of which 79 (11.1%) were heterozygous (-α3.7/αα) deletions and 1 (0.1%) homozy- gous (-α3.7/-α3.7). Ethnically, heterozygous deletions were higher (24.8%) in Afro-Brazilians. Comparison of hemato- logical parameters between individuals with normal genotype and those with heterozygous α+-thalassemia showed a statistically significant difference in the number of erythrocytes (p < 0.001), MCV (p < 0.001), MCH (p < 0.001) and Hb A2 (p = 0.007). This study is one of the first dedicated to investigating α-thalassemia 3.7 kb deletion in the population of the State Rio Grande do Norte state. Results obtained demonstrate the importance of investigating this condition in order to elucidate the causes of microcytosis and hypochromia.

βS haplotypes were studied in 47 non-related patients with sickle-cell anemia from the state of Rio Grande do Norte, Brazil. Molecular analysis was conducted by PCR/RFLP using restriction endonucleases XmnI, HindIII, HincII and HinfI to analyze six polymorphic sites from the beta cluster. Twenty-seven patients (57.5%) were identified with genotype CAR/CAR, 9 (19.1%) CAR/BEN, 6 (12.8%) CAR/CAM, 1 (2.1%) BEN/BEN, 2 (4.3%) CAR/Atp, 1 (2.1%) BEN/Atp and 1 (2.1%) with genotype Atp/Atp. The greater frequency of Cameroon haplotypes compared to other Brazilian states suggests the existence of a peculiarity of African origin in the state of Rio Grande do Norte.

35 unrelated individuals were studied for characterization as either heterozygous or homozygous for beta-thalassemia. Molecular analysis was done by PCR/RFLP to detect the mutations most commonly associated with beta-thalassemia (β0IVS-I-1, β+IVS-I-6, and β039). In the patients who showed none of these mutations, the beta-globin genes were sequenced. Of the 31 heterozygous patients, 13 (41.9%) had the β+IVS-I-6 mutation, 15 (48.4%) the β0IVS-I-1 mutation, 2 (6.5%) the β+IVS-I-110 mutation and 1 (3.2%) the β+IVS-I-5 mutation. IVS-I-6 was detected in the four homozygotes. The mutation in codon 39, often found in previous studies in Brazil, was not detected in the present case. This is the first study aiming at identifying mutations that determine beta-thalassemia in the state of Rio Grande do Norte.

La asociación del alelo betaS que produce hemoglobina S (HbS) y de un alelo de beta talasemia (betatal), ocurre principalmente en poblaciones que tienen simultáneamente ascendencia africana y mediterránea, y provoca un síndrome drepanocítico denominado HbS-beta talasemia. Este síndrome presenta una gran heterogeneidad clínica y genética debida en gran parte al alelo talasémico presente. Los individuos que portan un alelo talasémico beta0 presentan, en general, un curso clínico similar a los homocigotas para el alelo betaS de la HbS. En cambio, los que portan un alelo beta+ presentan un curso clínico variable, dependiendo de la reducción en la síntesis de cadena de beta globina. En este estudio se describe el caso clínico de dos pacientes con HbS-beta talasemia que consultaron en el Centro Hemato-Oncológico Pediátrico del Hospital Pereira Rossell. Se analizó el genotipo de los dos pacientes por medio de secuenciación automática del gen de la beta globina. Se demostró la presencia del alelo talasémico beta+ IVS-1-110 G->A en uno de los pacientes y del alelo beta0 codón39 C->T en el otro paciente, por lo cual ambos son heterocigotos compuestos betaS/betatal. Se discute la relación entre los datos clínicos y paraclínicos con los resultados del diagnóstico molecular. También se discute la importancia del diagnóstico molecular en relación con la composición y estructura de la población uruguaya.

Résumé L’association de l’allèle <FONT FACE=Symbol>b</FONT>S qui produit hémoglobine S (HbS) et d’un allèle de bêta tallasémie (<FONT FACE=Symbol>b</FONT>tal), arrive surtout chez des individus noirs d’Afrique et méditerranéens, et provoque un syndrome drépanocytaire appelé HbS-bêta tallasémie. Ce syndrome présente une grande hétérogé-néité clinique et génétique dûe surtout à l’allèle tallasémi-que présent. Les individus porteurs d’un allèle tallasémque <FONT FACE=Symbol>b</FONT>0 présentent en général,un cours clinique semblable aux homozygotes pour l’allèle <FONT FACE=Symbol>b</FONT>S de la HbS. En revanche, ceux qui portent un allèle <FONT FACE=Symbol>b</FONT>+ ont un cours clinique variable qui dépend de la réduction à la synthèse de la chaîne de beta globine. On décrit ici le cas de deux patients avec HbS-bêta tallasémie ayant consulté le Centre Hémato- Oncologique Pédiatrique de l’Hôpital Pereira Rossell. On analyse le génotype de deux patients au moyen de séquencement automatique du gène de bêta globine. On constate la présence de l’allèle tallasémique â+IVS-1-110G->A chez l’un des patients et de l’allèle <FONT FACE=Symbol>b</FONT>0 codon 39C->T chez l’autre. Les deux sont donc hétérozygotes composés <FONT FACE=Symbol>b</FONT>S/<FONT FACE=Symbol>b</FONT>tal. On discute ici le rapport entre les données cliniques et para cliniques et les résultats du diagnostic moléculaire.

Resumo A associação do alelo betaS que produz hemoglobina S (HbS) e de um alelo de beta talassemia betatal se dá principalmente em populações com ascendência africana e mediterrânea simultáneamente e provoca uma síndrome drepanocítica denominada HbS-beta talasemia. Esta síndrome apresenta uma grande heterogeneidade clínica e genética devida principalmente ao alelo para talassemia presente. Os indivíduos que são portadores de un alelo talassémico beta0 apresentam de maneira geral uma evolução clínica semelhante aos homozigotos para o alelo betaS da HbS. Por outro lado, os portadores do alelo beta+ apresentam um quadro clínico variável, dependendo da redução na síntese da cadeia beta-globina. Neste trabalho descrevemos os casos clínicos de dois pacientes com talassemia HbS.beta que foram admitidos no Centro Hemato-Oncológico Pediátrico do Hospital Pereira Rossel. O genotipo dos pacientes foi analizado pela sequenciação automática do gen da beta globina. A presença do alelo para talassemia beta+ IVS-1-110 G->A em um dos pacientes e do alelo beta0 codon39 C->T no outro foi demonstrada sendo os dois, por tanto, heterozi-gotos compostos betaS/betatal. Discute-se a relação entre os dados clínicos e paraclínicos com os resultados do diagnóstico molecular. Também se discute a importância do diagnóstico molecular em relação a composição e estrutura da população uruguaya.

Summary Association between allele betaS that produces hemoglobin S and allele (betatal) is mainly seen in African and Mediterranean populations, it causes the HbS-beta-thalassemia syndrome. This syndrome shows a wide range of clinical and genetic forms due mostly to the thalassemic allele. Carriers of thalassemic allele beta0 generally follow a similar clinical course to homocygote for talasémico betaS of the HbS. On the other hand, carriers of beta+ allele follow a variable clinical course, depending on the reduction of beta-globin chain. This study describe two clinical cases of patients with HbS-beta thalassemia of the Hematologic and Oncologic Centre of the Pereira Rossell Hospital. Genotypes of patients were analysed using automatic sequences of beta globin gen. Thalassemia allele beta+ IVS-1-110 G->A was seen in one patient and allele beta0 codon39 C->T in the other, so that both are compound heterozygous betaS/betatal. Clinical and paraclinical data is compared with molecular diagnostic findings; the importance of the molecular diagnosis in Uruguayan population is also discussed.

We report the clinical and laboratory findings concerning three unrelated Brazilian patients investigated for polycythemia, whose definitive diagnosis could only be established after the presence of Hb Coimbra (b99 Asp ® Glu) was demonstrated. This illustrates the importance of properly investigating hereditary hemoglobinopathies in cases of erythrocytosis because in some populations variants with high oxygen affinity may be more frequent than expected but go undetected when conventional electrophoresis is used as the sole detection procedure.

Hemoglobinopathies are the most common monogenic disorders worldwide; however, they have never been systematically studied from a genetic perspective in Uruguay. In this study, we determined the frequencies of hemoglobin variants in Afro-Uruguayans. A sample of 52 healthy unrelated Afro-Uruguayans from the northern (N = 28) and southern (N = 24) regions of the country was analyzed. Eight individuals (15.4%) were heterozygous for -alpha3,7thalassemia; seven of them (29.2%) were originally from the southern region, whereas one of them (3.6%) was from the northern region; the differences between both regions were statistically significant (p = 0.016 +/-0.003). The only structural mutation detected was betaS, which is typical of African populations. Four individuals (10%) were heterozygous for betaS, three of them (13.6%) from the South, and one (5.6%) from the North. The betaS haplotypes were analyzed in eight individuals: two were homozygous betaS/betaS, two were heterozygous betaS/betathal, and four were heterozygous betaS/betaª. This haplotype distribution (60% Bantu, 20% Benin, and 20% Bantu A2) is in agreement with historical records reporting a predominantly Bantu origin for the enslaved Africans brought to Uruguay. Even though this is a preliminary study, due to the small sample size, our results are suggestive of a relatively high incidence of hemoglobinopathies in the Afro-Uruguayan population.

OBJETIVO: Relacionar a função esplênica com os níveis do receptor solúvel da transferrina (sTfR) e com os valores da ferritina intra-eritrocitária (RCF) em pacientes com doenças falciformes. TIPO DE ESTUDO: Estudo prospectivo. LOCAL: Hospital das Clínicas da Faculdade de Ciências Médicas da UNICAMP, atendimento terciário. PARTICIPANTES: 60 pacientes portadores de doenças falciformes, na fase estável da doença, sem transfusão de sangue nos últimos 3 meses; 28 indivíduos normais, sem sinais clínicos e laboratoriais de anemia. VARIÁVEIS ESTUDADAS: Determinação do ferro sérico, capacidade de ligação do ferro à transferrina, ferritina sérica, RCFe sTfR. Avaliação da função esplênica: quantificação de hemácias com "pits". RESULTADOS: Pacientes com anemia falciforme: os níveis de sTfR foram significativamente mais elevados do que nos indivíduos normais e com Hb SC (p=0,0001); correlação inversa entre sTfR e Hb Fetal (p=0,0016); valores de RCF significativamente mais elevados do que nos indivíduos normais e pacientes SC (p=0,0001); correlação entre RCF e hemácias com "pits" (p=0,0512).CONCLUSÕES: A associação entre sTfR e Hb Fetal confirma a contribuição da Hb Fetal na melhora do quadro hemolítico, o que, conseqüentemente, minimiza a reativação da expansão eritróide. Os altos níveis de sTfR observados nos pacientes com doença falciforme não estão relacionados com o grau de hipofunção esplênica. A deficiência do processo de exocitose exercido pelo baço pode contribuir para o acúmulo de RCF nos pacientes com anemia falciforme.

OBJECTIVE: To correlate spleen function with soluble transferrin receptor (sTfR) levels and red cell ferritin (RCF) values in patients with sickle cell diseases. DESIGN: Prospective study. LOCATION: University Hospital, School of Medical Sciences, State University of Campinas; a tertiary hospital. PARTICIPANTS: 60 patients with sickle cell diseases, in a steady state, who had not received blood transfusions for 3 months; 28 normal individuals with no clinical or laboratory signs of anemia. MEASUREMENTS: Determination of serum iron, transferrin iron-binding capacity, serum ferritin, RCF and sTfR. Evaluation of spleen function: erythrocytes with pits were quantified. RESULTS: Patients with sickle cell anemia had sTfR levels significantly higher than in normal individuals or those with HbSC (p=0.0001) and there was an inverse correlation between sTfR and fetal Hb (p=0.0016). RCF values were significantly higher in sickle cell anemia patients than in normal individuals or those with HbSC (p=0.0001), and there was a correlation between RCF and pitted erythrocytes (p=0.0512). CONCLUSION: The association between sTfR and fetal Hb confirms the contribution of fetal Hb to improving the hemolytic state by minimizing the consequent reactive erythrocyte expansion. High sTfR levels are not related to the degree of spleen function deficiency seen in sickle cell disease patients. The deficiency in the exocytosis process of the spleen occurring in sickle cell anemia patients may contribute to their accumulation of RCF.

Nós caracterizamos a base molecular da talassemia b em uma paciente negra brasileira, de 8 anos de idade, com um quadro clínico extremamente benigno de S-b+ talassemia. A paciente tinha uma hemoglobina total de 10,1 g/dl, com 57% de HbS. O sequenciamento direto do DNA, amplificado por PCR, revelou que a paciente é heterozigota da mutação b+ IVS-I-6 (T®C). Esta mutação, algumas vezes referida como o tipo português de talassemia, foi encontrada em associação com o polimorfismo C®T no codon 2 do gene b e, de nosso conhecimento, não havia sido previamente descrita na população negra brasileira.

We report on an eight-year-old Brazilian girl with S-b+ thalassemia. The patient had a steady 10.1 g/dl hemoglobin with 57% HbS. Direct sequence analysis of b-globin gene showed her to be heterozygous for the IVS-I-6 (T®C) mutation. This b+ thalassemia mutation, sometimes referred to as the Portuguese type, was found to be associated with the C®T polymorphism at codon 2. In combination with the bS gene, this mutation results in very mild sickle cell disease symptoms.