Α-Thalassemia: Genotypic Profile Associated with Ethnicity and Hematological Differentiation of Iron Deficiency Anemia in the Region of Uberaba, Minas Gerais, Brazil.

α-Thalassemia (α-thal) is a hereditary hemoglobinopathy characterized by microcytic anemia due to impaired production of α chains of human globin. Brazilian studies show that the most common genotype is an -α(3.7) deletion with the loss of one or two α genes. As the production of α chains is not as accentuated in these cases, the correct diagnosis can only be achieved through molecular analysis that is not usually routinely performed by laboratories. We investigated the occurrence of α-thal babies born between September 2011 to January 2013 at the hospital of the Universidade Federal do Triângulo Mineiro (UFTM), Uberaba, Brazil, and blood donors of the Uberaba Regional Blood Center, Hemominas Foundation, Uberaba, Brazil, correlating it with ethnicity and differences between hematological parameters of donors, α-thal and iron deficiency patients. α-Thalassemia was investigated for the most common deleted alleles (-α(3.7), -α(4.2), - -(SEA), - -(FIL), - -(THAI), -(α)(20.5) and - -(MED)). The incidence in newborns was 13.16% with a predominance of heterozygosity for the -α(3.7) genotype (12.35%), followed by the -α(3.7)/-α(3.7) (0.46%) and αα/-α(4.2) genotypes (0.35%). In blood donors, the prevalence of α-thal was 14.89%, with all cases being heterozygous for the -α(3.7) deletion. There was an association of the α-thal genotype with African ancestors for both groups, thereby confirming published data and showing the strong influence of Blacks on the composition of the population of Brazil's southeastern region. Minor changes were found between hematological parameters of blood donors with iron deficiency and α-thal that did not contribute to the differential diagnosis between the two types of anemia.

DBT- and DBTO2-induced dysplasia and their associated proteomic alterations in the small intestines of Wistar rats.

Dibenzothiophene (DBT) and its oxidized derivative dibenzothiophene sulfone (DBTO2) are important representatives of polycyclic aromatic hydrocarbons (PAHs). Due to the importance of PAHs in oncogenesis and the lack of toxicological investigations related to DBT and DBTO2, this work proposes to assess their toxic and molecular effects caused by chronic treatment of Wistar rats. In parallel, their effects were compared to those caused by treatment with 1,2-dimethylhydrazine (DMH), a classic mutagenic agent. At the 14th day post-treatment, the animals were sacrificed and blood withdrawn for hematology and evaluation of liver and pancreatic functions. No significant alterations were observed. Nevertheless, histopathological analyses revealed dysplastic lesions in the intestines of animals treated with DBT and DBTO2. CD44 and carcinoembryonic antigen (CEA) staining demonstrated an approximately 3-fold increase in expression of both tissue markers for animals administered DBT, DBTO2, and DMH. A comparative two-dimensional gel analysis revealed additional 23 proteins exhibiting altered levels in the small intestines caused by exposure to DBT and DBTO2. At last, a protein-metabolite interaction map provided major insights into the metabolism of the dysplastic tissues. Our results provided strong evidence that DBT and its derivative could potentially act as cancer inducers, highlighting their toxicological and environmental relevance.

Coinheritance of α-thalassemia decreases the risk of cerebrovascular disease in a cohort of children with sickle cell anemia.

The study estimated α-thalassemia (α-thal) prevalence and assessed its associations with clinical and hematological features in a random sample of Brazilian children with sickle cell anemia (208 Hb SS and 13 Hb S-ß°-thal). α-Thalassemia genotyping was carried out by multiplex polymerase chain reaction (m-PCR) for seven alleles. Clinical and hematological data were retrieved from the 221 children's medical files. Their ages ranged from 2.5 to 10.4 years. Of the Hb SS children, 27.9% carried -α(3.7)/αα and 1.4% -α(3.7)/-α(3.7). The presence of α-thal was significantly associated with reduction in MCV, MCH, WBC values and reticulocyte counts. No significant association with blood transfusion or acute chest syndrome (ACS), was found. α-Thalassemia genotypes were strongly associated with reduction in risk for cerebrovascular disease (CVD) (conditional and abnormal transcranial Doppler or stroke; p = 0.007). The interaction of α-thal with other modulating factors should be investigated in order to define subphenotypes of the disease and to use them as clinical tools in the follow-up care of patients.

ß-globin gene cluster haplotypes in a cohort of 221 children with sickle cell anemia or Sß°-thalassemia and their association with clinical and hematological features.

BACKGROUND/AIMS: ß(S)-Haplotype prevalence and its associations with clinical and hematological characteristics were assessed in Brazilian children with sickle cell anemia or Sß°-thalassemia. METHODS: A retrospective randomized cohort study was undertaken with 208 SS and 13 Sß°-thalassemia children derived from the Newborn Screening Program of the state of Minas Gerais. ß(S)-Haplotypes were determined by PCR-RFLP. RESULTS: Thirty-nine percent of the SS subjects had the CAR/CAR genotype, 33% had CAR/Ben, 24% had Ben/Ben, 1% had CAR/Atp, 1% had Ben/Atp, and 1% had Arab-Indian/Ben; 1% could not be characterized. Of the Sß°-thalassemia children, 5 were CAR/undefined, 2 were Ben/undefined, and 1 was CAM/undefined. There was no significant association between ß(S)-haplotypes and the total Hb, Hb F, MCV, MCH, WBC, and reticulocyte count among the SS children. Likewise, no significant association was detected between ß(S)-haplotypes and the frequency of acute chest syndrome episodes, blood transfusions, splenic sequestration, or cerebrovascular disease (high-risk/conditional transcranial Doppler ultrasonography or clinical stroke). A limited number of Sß°-thalassemia children precluded valid analyses. CONCLUSIONS: The prevalence of ß(S)-haplotypes in this study is in agreement with the historical records of African slaves brought to the state of Minas Gerais. Furthermore, ß(S)-haplotypes CAR and Ben were not associated with any analyzed feature of children with sickle cell anemia.

Production of specific polyclonal antibodies anti-human factor IX

In this work, polyclonal antibodies anti-human Factor IX were produced in New Zealand rabbits by immunization with commercial pure human FIX (hFIX) (Octanyne®, Octapharma, USA). The serum containing immunoglobulins anti-hFIX was useful to detect hFIX antigen in human plasma fractions submitted to anionic exchange chromatographic process and with a large yield. Immunoassays (ELISA) using bovine serum albumin, trypsin and peptides generated by cleavage assays with trypsin as digestion enzyme was performed and revealed adequate specificity of the polyclonal antibodies produced.

Neste trabalho foram produzidos anticorpos policlonais anti-fator IX humano em coelhos New Zealand imunizados com FIX humano (hFIX) comercial puro (Octanyne®, Octapharma, EUA). O soro contendo as imunoglobulinas anti-hFIX foi útil para a detecção do antígeno hFIX em frações do plasma humano submetido a cromatografia de troca iônica. Imunoensaios (ELISA) usando soro-albumina bovina, tripsina e peptídeos gerados por ensaios de clivagem com tripsina com enzima de digestão foram realizados e revelaram especificidade adequada dos anticorpos policlonais produzidos.