Association of FOXO3 polymorphism (rs3800231) and clinical subphenotypes of beta thalassemic individuals

Abstract Introduction Studies have shown that the loss of the FOXO3 transcriptional function is involved in the pathophysiology of some chronic erythroid disorders, including beta-thalassemia (β-thal). Therefore, the single nucleotide polymorphism (SNP) rs3800231 (35-2764A > G) could contribute to alterations in its transcriptional activity, acting as a modifier of β-thal phenotypic manifestations. Objective and method In order to better understand the genotypic and/or allelic distributions among β-thal patients, we evaluated 83 β-thal heterozygous and 20 homozygous, compared to 117 individuals without hemoglobinopathies (control group). Additionally, we verified any influence of the FOXO3 polymorphism on clinical manifestations among β-thal homozygotes. Results We obtained higher frequencies of the wild-type homozygous (AA) and the wild-type allele (A) in the β-thal group (p< 0.0001 and p= 0.00014, respectively). The most common clinical manifestations found among β-thal homozygotes were iron overload (90%), splenomegaly (65%) and bone complications (35%), e.g., osteopenia/osteoporosis. We observed that close to 80% of the patients presenting such manifestations had the genotype AA. However, we did not find any significant involvement of the FOXO3 polymorphism in clinical manifestation occurrences. Conclusion Thus, we concluded that the SNP rs3800231 did not play a significant role as a modifier of the clinical manifestations observed in the β-thal homozygotes studied.

A interação entre Hb B2, Hb S e beta talassemia: um relato de caso familiar

As hemoglobinopatias são doenças hereditárias que incluem talassemia e doença falciforme. O objetivo do presente estudo foi destacar a interação entre a variante Hb S, formas talassêmicas (beta talassemia por IVS1-6) e a variante Hb B2. Os exames realizados foram hemograma completo, eletroforese de hemoglobina em pH ácido e alcalino, dosagem de hemoglobina A2, Cromatografia Líquida de Alta Performance e investigação molecular. Esse relato evidencia a interação entre os polimorfismos de hemoglobina na população brasileira e a necessidade de adequada interpretação dos resultados de testes clássicos para a melhor compreensão dos casos.

Hemoglobinopathies are a hereditary disease that includes thalassemia and sickle cell disease. The present study aimed to show the interaction between the Hb S variant, thalassemia forms (Beta-thalassemia by IVS1-6), and the Hb B2 variant. The tests performed were complete blood count, hemoglobin electrophoresis at acid and alkaline pH, hemoglobin A2 dosage, High-Performance Liquid Chromatography, and molecular investigation. This report highlights the interaction between hemoglobin polymorphisms in the Brazilian population and the need for an adequate interpretation of the results of classical tests for a better understanding of the cases.

Association of HMIP1 C-893A polymorphism and disease severity in patients with sickle cell anemia

Abstract Introduction: Sickle cell anemia (SCA) is a Mendelian disorder with a heterogeneous clinical course. The reasons for this phenotypic diversity are not entirely established, but it is known that high fetal hemoglobin levels lead to a milder course of the disease. Additionally, genetic variants in the intergenic region HBS1L-MYB promote high levels of fetal hemoglobin into adulthood. Objective: In the present study, we investigated the HMIP1 C-839A (rs9376092) polymorphism, located at the HBS1L-MYB intergenic region block 1, in SCA patients. Method: We analyzed 299 SCA patients followed in two reference centers in Brazil. The HMIP1 C-839A (rs9376092) genotypes were determined by allele specific polymerase chain reactions. Clinical and laboratory data were obtained from patient interviews and medical records. Results: The median fetal hemoglobin levels were higher in patients with the HMIP1 C-839A (rs9376092) AA genotype (CC = 6.4%, CA = 5.6% and AA = 8.6%), but this difference did not reach significance (p = 0.194). No association between HMIP1 C-839A (rs9376092) genotypes and other clinical and laboratorial features was detected (p > 0.05). Conclusion: In summary, our data could not support the previously related association between the HMIP1 C-893A (rs9376092) polymorphism and differential fetal hemoglobin levels.

The relationship between of ACE I/D and the MTHFR C677T polymorphisms in the pathophysiology of type 2 diabetes mellitus in a population of Brazilian obese patients

ABSTRACT Objectives This study aimed to evaluate the frequencies of the angiotensin converting enzyme (ACE) gene insertion/deletion (I/D) and methylenetetrahydrofolate reductase (MTHFR) gene C677T polymorphisms in obese patients with and without type 2 diabetes mellitus (T2DM). Subjects and methods These polymorphisms were analyzed by polymerase chain reaction in 125 patients with obesity, 47 (T2DM) and 78 (Control Group). Results No significant difference was found on comparing the T2DM and Control Groups in respect to the genotypic frequencies of the polymorphisms - (II: 13.3% vs. 12.0%; ID: 37.8% vs. 37.3; DD: 48.9% vs. 50.7%; CC: 36.2% vs. 39.0%; CT: 46.8% vs. 49.3%; TT: 17.0% vs. 11.7%), and alleles (I: 32.2% vs. 30.7%; D: 67.8% vs. 69.3%; C: 59.6% vs. 63.6%; T: 40.4% vs. 36.4%) and their synergisms in the pathophysiology of T2DM. On analyzing the T2DM Group, there were no significant differences in the presence of complications. In this population of Brazilian obese patients, no correlation was found between the ACE and MTHFR polymorphisms in the development of T2DM. Conclusion Analyzing only the group with diabetes, there was also no relationship between these polymorphisms and comorbidities.

O processo de consentimento livre e esclarecido nas pesquisas em doença falciforme / The process of informed consent in research on sickle cell disease / El proceso de consentimiento informado en investigaciones sobre la enfermedad de células falciformes

Doença falciforme diz respeito a grupo de hemoglobinopatias associadas à presença da hemoglobina S. Sendo majoritária na população negra, e acometendo em sua maioria vulneráveis e vulnerados, a forma homozigota da doença ­ a anemia falciforme ­ é considerada relevante problema de saúde pública no Brasil. Entendendo a pesquisa científica como essencial para a promoção da saúde e para melhorar a qualidade de vida dos pacientes, o processo de consentimento livre e esclarecido deve ser aplicado para superar, na medida do possível, vulnerabilidades a que as pessoas com doença falciforme estão expostas. Recursos lúdicos, transmissão coletiva de informações, proteção conferida pelas associações de pacientes e a formação permanente em ética em pesquisa por parte dos profissionais que aplicam o consentimento são indicados como ferramentas para otimizar esse processo.

Abstract Sickle cell disease is a term that refers to a group of hemoglobinopathies associated with the presence of hemoglobin S. Occurring primarily in black population, and affecting mainly the most vulnerable, the homozygous form of the disease, sickle cell anemia, is considered an important public health problem in Brazil. Understanding scientific research as essential to promote health and improve the quality of life of people with sickle cell disease, the informed consent process should be done to overcome, as far as possible, the vulnerabilities which people with sickle cell disease are exposed to. The use of recreational resources, the transmission of collective information, the protection provided by patient associations and the continuous training of ethics in research by the professionals involved in the agreement are shown as tools for the optimization of this process.

Resumen La enfermedad de células falciformes es un término que se refiere a un grupo de hemoglobinopatías asociado con la presencia de la hemoglobina S. La forma homocigótica del gen de la enfermedad se presenta sobre todo en la población negra y se considera un problema importante de salud pública en Brasil, que afecta sobre todo a vulnerables y vulnerados. Entendiendo a la investigación científica como esencial para la promoción de la salud y para el aumento de la calidad de vida de las personas con enfermedad de células falciformes, el proceso de consentimiento libre e informado debe ser aplicado para superar, en la medida de lo posible, las vulnerabilidades a las que las personas con la enfermedad de células falciformes están expuestas. El uso de los recursos recreativos, la transmisión de información colectiva, la protección que ofrecen las asociaciones de pacientes y la formación continua en ética en investigación por parte de los profesionales que aplican el consentimiento son señaladas como herramientas para la optimización de este proceso.

Hemoglobin D-Punjab: origin, distribution and laboratory diagnosis

This review discusses hemoglobin D-Punjab, also known as hemoglobin D-Los Angeles, one of the most common hemoglobin variants worldwide. It is derived from a point mutation in the beta-globin gene (HBB: c.364G>C; rs33946267) prevalent in the Punjab region, North-western Indian. Hemoglobin D-Punjab can be inherited in heterozygosis with hemoglobin A causing no clinical or hematological alterations, or in homozygosis, the rarest form of inheritance, a condition that is commonly not related to clinical symptomatology. Moreover, this variant can exist in association with other hemoglobinopathies, such as thalassemias; the most noticeable clinical alterations occur when hemoglobin D-Punjab is associated to hemoglobin S. The clinical manifestations of this association can be similar to homozygosis for hemoglobin S. Although hemoglobin D-Punjab is a common variant globally with clinical importance especially in cases of double heterozygosis, hemoglobin S/D-Punjab is still understudied. In Brazil, for example, hemoglobin D-Punjab is the third most common hemoglobin variant. Thus, this paper summarizes information about the origin, geographic distribution, characterization and occurrence of hemoglobin D-Punjab haplotypes to try to improve our knowledge of this variant. Moreover, a list of the main techniques used in its identification is provided emphasizing the importance of complementary molecular analysis for accurate diagnosis.

Tratamento de folículo anovulatório persistente em égua: relato de caso / Treatment of persistent anovulatory follicle in mare: a case report

O presente estudo busca apresentar o processo diagnóstico, evolução e tratamento, bem como registrar um caso de folículo anovulatório persistente na espécie equina. Para tanto, realizou-se atendimento clínico em uma égua da raça Quarto de Milha de nove anos de idade, no Hospital Veterinário da Universidade Paulista de São José dos Campos, com histórico de infertilidade. Foi realizada a avaliação clínica do paciente e exame ultrassonográfico do sistema reprodutivo, utilizando o equipamento Ultrassom Chison D600 com probe linear 7,5MHz no qual foi identificada estrutura compatível com folículo anovulatório no ovário direito. O tratamento proposto seria a utilização 5000 UI de gonadotropina coriônica humana (hCG - Vetecor - Hertape Calier) por via intravenosa e 250 mg de Cloprostenol (Prolise® -Tecnopec), por via intramuscular, no nono dia após a aplicação do hCG. No entanto por apresentar quadro de desconforto abdominal 24 horas após a aplicação do hCG, o tratamento foi suspenso a pedido do proprietário, não sendo realizada a indução da luteólise. O animal foi reavaliado com novo exame ultrassonográfico após sete dias da aplicação do hCG, onde foram observadas imagens ultrassonográficas compatíveis com a ovulação do folículo. A terapia proposta foi eficiente para resolução do folículo anovulatório, no entanto, a origem do quadro de desconforto abdominal pode ser evidenciada, sugerindo que maiores esforços sejam necessários para investigar o efeito do tratamento na saúde do animal...

This study seeks to make the diagnostic process, evolution and treatment, as well as to register a case of persistent anovulatory follicle in the equine species. Therefore, a mare of the Quarter Horse race of nine-year-old with a history of infertility was examined at the Veterinary Hospital of Universidade Paulista in São José dos Campos. It was performed a clinical assessment of the patient and ultrasound examination of the reproductive system, using ultrasound equipment Chison D600 with 7.5 MHz linear probe. It was identified a structure compatible with anovulatory follicle in the right ovary. The proposed treatment would be to use 5000 IU IV human chorionic gonadotropin (hCG - Vetecor - Hertape Calier) and 250 mg Cloprostenol IM (Prolise® - Tecnopec) on the ninth day after application of hCG. However, the animal showed abdominal discomfort 24 hours after application of hCG, so the treatment was suspended at the request of the owner, has not been accomplished to induce luteolysis. The animal was reassessed with new ultrasound seven days after application of hCG, which were seen ultrasound images compatible with ovulation follicle. The proposed therapy was effective for solving the anovulatory follicle, however, it was evident abdominal discomfort, suggesting that further efforts are necessary to investigate the treatment effect on the health of the animal...

Oxidative stress and antioxidant status in beta-thalassemia heterozygotes

Background: Several studies have evaluated the oxidant and antioxidant status of thalassemia patients but most focused mainly on the severe and intermediate states of the disease. Moreover, the oxidative status has not been evaluated for the different beta-thalassemia mutations. Objective: To evaluate lipid peroxidation and Trolox equivalent antioxidant capacity in relation to serum iron and ferritin in beta thalassemia resulting from two different mutations (CD39 and IVS-I-110) compared to individuals without beta-thalassemia. Methods: One hundred and thirty subjects were studied, including 49 who were heterozygous for beta-thalassemia and 81 controls. Blood samples were subjected to screening tests for hemoglobin. Allele-specific polymerase chain reaction was used to confirm mutations for beta-thalassemia, an analysis of thiobarbituric acid reactive species was used to determine lipid peroxidation, and Trolox equivalent antioxidant capacity evaluations were performed. The heterozygous beta-thalassemia group was also evaluated for serum iron and ferritin status. Results: Thiobarbituric acid reactive species (486.24 ± 119.64 ng/mL) and Trolox equivalent antioxidant capacity values (2.23 ± 0.11 mM/L) were higher in beta-thalassemia heterozygotes compared to controls (260.86 ± 92.40 ng/mL and 2.12 ± 0.10 mM/L, respectively; p-value < 0.01). Increased thiobarbituric acid reactive species values were observed in subjects with the CD39 mutation compared with those with the IVS-I-110 mutation (529.94 ± 115.60 ng/mL and 453.39 ± 121.10 ng/mL, respectively; p-value = 0.04). However, average Trolox equivalent antioxidant capacity values were similar for both mutations (2.20 ± 0.08 mM/L and 2.23 ± 0.12 mM/L, respectively; p-value = 0.39). There was no influence of serum iron and ferritin levels on thiobarbituric acid reactive species and Trolox equivalent antioxidant capacity values. Conclusion: ...

The influence of hydroxyurea on oxidative stress in sickle cell anemia

OBJECTIVE: The oxidative stress in 20 sickle cell anemia patients taking hydroxyurea and 13 sickle cell anemia patients who did not take hydroxyurea was compared with a control group of 96 individuals without any hemoglobinopathy. METHODS: Oxidative stress was assessed by thiobarbituric acid reactive species production, the Trolox-equivalent antioxidant capacity and plasma glutathione levels. RESULTS: Thiobarbituric acid reactive species values were higher in patients without specific medication, followed by patients taking hydroxyurea and the Control Group (p < 0.0001). The antioxidant capacity was higher in patients taking hydroxyurea and lower in the Control Group (p = 0.0002 for Trolox-equivalent antioxidant capacity and p < 0.0292 for plasma glutathione). Thiobarbituric acid reactive species levels were correlated with higher hemoglobin S levels (r = 0.55; p = 0.0040) and lower hemoglobin F concentrations(r = -0.52; p = 0.0067). On the other hand, plasma glutathione levels were negatively correlated with hemoglobin S levels (r = -0.49; p = 0.0111) and positively associated with hemoglobin F values (r = 0.56; p = 0.0031). CONCLUSION: Sickle cell anemia patients have high oxidative stress and, conversely, increased antioxidant activity. The increase in hemoglobin F levels provided by hydroxyurea and its antioxidant action may explain the reduction in lipid peroxidation and increased antioxidant defenses in these individuals.

What influences Hb fetal production in adulthood?: [review]

Human hemoglobin genes are located in α and β globin gene clusters in chromosomes 16 and 11, respectively. Different types of hemoglobin are synthesized according to the stage of development with fetal hemoglobin (α2γ2) (Hb F) being the main hemoglobin in the fetal period. After birth, there is a reduction (to about 1 percent) in Hb F levels and adult hemoglobin, Hb A (2α2β2), increases to more than 96 percent of total hemoglobin. However, some genetic conditions whether linked to the β-globin gene cluster or not are associated with high Hb F levels in adults. Among those linked to β-globin are hereditary persistence of fetal hemoglobin, delta-beta thalassemia (δβ-Thalassemia) and the XmnI polymorphism (-158 C > T). Other polymorphisms not related to β-globin gene cluster are known to influence the γ-globin gene expression in adulthood. The most relevant polymorphisms that increase concentrations of Hb F are the HMIP locus on chromosome 6, the BCL11A locus on chromosome 2, the Xp22.2 region of the X chromosome and the 8q region on chromosome 8. Findings from our research group studying genetic factors involved in γ-globin gene regulation in adults without anemia in the northwestern region of São Paulo State showed that high Hb F levels are influenced by the presence of hereditary persistence of fetal hemoglobin mutations and the XmnI polymorphism, suggesting that both genetic alterations characterize the molecular basis of the evaluated population.

Perfil de beta talassemia heterozigota obtido a partir de análise data mining em banco de dados: [carta ao editor] / The profile of beta thalassemia obtained by data mining analysis in a database: [letter to editor]

Variations in the phenotypic expression of heterozygous beta thalassemia reflect the formation of different populations. To better understand the profile of heterozygous beta-thalassemia of the Brazilian population, we aimed at establishing parameters to direct the diagnosis of carriers and calculate the frequency from information stored in an electronic database. Using a Data Mining tool, we evaluated information on 10,960 blood samples deposited in a relational database. Over the years, improved diagnostic technology has facilitated the elucidation of suspected beta thalassemia heterozygote cases with an average frequency of 3.5 percent of referred cases. We also found that the Brazilian beta thalassemia trait has classic increases of Hb A2 and Hb F (60 percent), mainly caused by mutations in beta zero thalassemia, especially in the southeast of the country.