First Report of ß-Thalassemia Intermedia in a Patient Compound Heterozygous for -92 (C>T) and Codons 36/37 (-T) Mutations.

The -92 (C>T) (HBB: c.-142C>T) is a silent ß-thalassemia (ß-thal) mutation previously described in combination with several ß0 mutations and expressed as ß-thal intermedia (ß-TI). Heterozygous individuals are known to be completely asymptomatic showing borderline hemoglobin (Hb), mean corpuscular volume (MCV) and Hb A2 levels. Here, we report the first incidence of -92 in Eastern Europe and in Azerbaijan, and the first case in combination with codons 36/37 (-T) (HBB: c.112delT) mutation.

Molecular and geographical heterogeneity of hemoglobinopathy mutations in Azerbaijanian populations.

With the carrier rate of 4%-8.6%, ß-thalassemia is one of the most prevalent hereditary disorders in Azerbaijan. Taking into consideration the high frequency of ß-thalassemia as well as the occurrences of several other hemoglobinopathies, we conducted a large genotyping study to investigate the mutational background of common hemoglobinopathies in the country. Α- and ß-globin genes were evaluated in the carriers of mutations identified via hematological indices and hemoglobin fractions (n = 1,757). Genotyping of ß-thalassemia carriers identified through population screening revealed 32 mutations, with codon 8 [-AA]-34.96%, IVS-II-1 [G > A]-16.35%, and IVS-I-110 [G > A]-10.12% leading the spectrum. Analysis of associations of ß-thalassemia mutations with geographical regions of the country identified the strongest association between codon 8 [-AA] and Shaki-Zaqatala, and codon 5 [-CT] in Mountainous Shirvan regions (ri > 6.00; p < 0.05). HbS, HbD-Punjab, and HbE were the most prevalent among our variant hemoglobin cohort, commonly inherited in compounds with ß-thalassemia than in the homozygous state. We identified nine α-thalassemia mutations, 20.5 kb and 3.7 kb deletions together accounting for 74% of the spectrum. Point mutations of α-thalassemia were less common among our observations and were mainly inherited in compounds with deletions. Our results allow a better understanding of the wide spectrum of mutations in Azerbaijan and highlights the high heterogeneity of hemoglobinopathies in the local population.

A Novel Frameshift Mutation at Codon 2 (-T) (HBB: c.9delT) and First Report of Three New ß-Globin Mutations From Azerbaijan.

We identified a novel mutation of ß-thalassemia (ß-thal) in a heterozygous carrier from Azerbaijan. Phenotypical data and molecular mechanisms of codon 2 (-T) (HBB: c.9delT) was relevant to ß0-thal. Additionally, we here report two new mutations on the HBB gene, not observed previously, in the local population as well as a non causative promoter mutation -198 (A>G) (HBB: c.-248A>G).

Thalassemia in the laboratory: pearls, pitfalls, and promises.

Thalassemia is one of the most common hereditary disorders of the developing world, and it is associated with severe anemia and transfusion dependence. The global health burden of thalassemia has increased as a result of human mobility and migration in recent years. Depending on inherited mutations, thalassemia patients exhibit distorted hemoglobin (Hb) patterns and deviated red cell indices, both of which can be used to support identification by diagnostic tools. Diagnostic approaches vary depending on the target population and the aim of the testing. Current methods, which are based on Hb patterns, are used for first-line screening, whereas molecular testing is needed for conformation of the results and for prenatal and preimplantation genetic diagnosis. In the present paper, we review the diagnostic parameters, pitfalls, interfering factors, and methods; currently available best-practice guidelines; quality assurance and standardization of the procedures; and promising laboratory technologies for the future of thalassemia diagnosis.

Codon 14 (+T) (HBB: c.44_45insT): a Rare ß-Thalassemia Mutation Reported Only in Azerbaijan.

Codon 14 (+T) (HBB: c.44_45insT) is a very rare ß-thalassemia (ß-thal) mutation previously reported in three ß-thal major (ß-TM) patients of Azerbaijani origin. None of the previous reports described the genotype-phenotype correlation of the mutation. We here report the first case of homozygous codon 14 together with data of the heterozygous parents.

Spectrum of BCR-ABL mutations in Azerbaijanian imatinib-resistant patients with chronic myeloid leukemia.

Objective: BCR-ABL1 kinase domain (KD) mutations can lead to resistance to first- and second-generation tyrosine kinase inhibitors (TKIs) in chronic myeloid leukemia (CML). Here, we present the first report of the spectrum of mutations in the BCR-ABL1 KD of CML patients from Azerbaijan. Materials and methods: Samples for mutation screening were obtained from patients experiencing resistance to first line TKIs or from patients in acceleration phase (AP) or blast crisis (BC) at the time of diagnosis. The cDNA region corresponding to BCR-ABL1 KD was sequenced by pyrosequencing method. The χ2 test was used to assess the association of categorical variables between mutation-positive and -negative groups. In addition, the Kaplan-Meier method was applied to generate survival curves. Results: Eight different point mutations were identified in 22 (13.4%) out of 163 CML patients experiencing resistance to TKIs. The types of mutations detected were as follows: Contact binding site mutations 50% (11), SH2 domain mutations 27.4% (six), P-loop mutations 18.1% (four), and SH3 domain mutations accounting for 4.5% (one). The most common mutation was T315I, accounting for 5% (n = 8) of all patients. Significant association was identified between BCR-ABL1 mutations and additional chromosomal aberrations as well as between the mutations and disease phases (p < 0.05). Twelve out of 22 patients with BCR-ABL1 mutations and seven out of eight with T315I were in BC. Overall survival (OS) of the patients with BCR-ABL1 mutations was significantly lower comparing to the patients with no mutation (p < 0.05) and 8 patients with T315I mutation presented OS of 0%. Conclusion: T315I was the most commonly identified BCR-ABL1 mutation in TKI-resistant CML patients of Azerbaijani origin, being associated with disease progression and poor OS.

Diagnostic accuracy of fine needle aspiration biopsy versus postoperative histopathology for diagnosing thyroid malignancy.

INTRODUCTION: Ultrasound-guided fine needle aspiration biopsy (FNAB) is currently widely used for the initial screening of patients with thyroid nodules enabling prevention of unnecessary surgery. The purpose of this study was to retrospectively analyse the diagnostic accuracy of thyroid FNAB compared with postoperative histopathology of a large cohort from Azerbaijan. METHODS: We evaluated the FNAB results of 738 patients who underwent thyroid surgery at the National Centre of Oncology in Azerbaijan. The measures of diagnostic accuracy were calculated for the ultrasound-guided preoperative FNAB results (based on the six diagnostic categories of the Bethesda classification) compared with postoperative histopathologic results (benign or malignant) for correspondent areas. RESULTS: Considering both DC V and DC VI categories (387 cases) as 'cytologic-positive' and DC II category (72 cases) as 'cytologic-negative', we found 14 false-positive and 10 false-negative results. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and diagnostic accuracy were 97.4%, 86.1%, 96.4%, 81.6% and 94.8%, respectively. Conversely, when considering only the DC VI category as 'cytologic-positive', the sensitivity, specificity, PPV, NPV and diagnostic accuracy of FNA were 93.2%, 100%, 100%, 81.6% and 97.1%, respectively. CONCLUSIONS: The results of our cohort demonstrated high levels of diagnostic accuracy, supporting FNAB's role as a reliable diagnostic tool in the preoperative evaluation of thyroid nodules. The sensitivity, specificity, NPV, PPV and accuracy of thyroid FNAB in our institution were comparable with those of other institutions.

ß-Thalassemia intermedia: a comprehensive overview and novel approaches.

ß-Thalassemia intermedia is a clinical condition of intermediate gravity between ß-thalassemia minor, the asymptomatic carrier, and ß-thalassemia major, the transfusion-dependent severe anemia. It is characterized by a significant clinical polymorphism, which is attributable to its genetic heterogeneity. Ineffective erythropoiesis, chronic anemia, and iron overload contribute to the clinical complications of thalassemia intermedia through stepwise pathophysiological mechanisms. These complications, including splenomegaly, extramedullary erythropoiesis, iron accumulation, leg ulcers, thrombophilia, and bone abnormalities can be managed via fetal hemoglobin induction, occasional transfusions, chelation, and in some cases, stem cell transplantation. Given its clinical diversity, thalassemia intermedia patients require tailored approaches to therapy. Here we present an overview and novel approaches to the genetic basis, pathophysiological mechanisms, clinical complications, and optimal management of thalassemia intermedia.

Thiopurine S-Methyltransferase as a Pharmacogenetic Biomarker: Significance of Testing and Review of Major Methods.

BACKGROUND: Thiopurine S-methyltransferase (TPMT) enzyme metabolizes thiopurine drugs which are widely used in various disciplines as well as in leukemias. Individual enzyme activity varies depending on the genetic polymorphisms of TPMT gene located at chromosome 6. Up to 14% of population is known to have a decreased enzyme activity, and if treated with standard doses of thiopurines, these individuals are at a high risk of severe Adverse Drug Reactions (ADR) as myelosuppression, gastrointestinal intolerance, pancreatitis and hypersensitivity. However, TPMT-deficient patients can successfully be treated with decreased thiopurine doses if enzyme status is identified by a prior testing. TPMT status identification is a pioneering experience in application of pharmacogenetic testing in clinical settings. 4 TPMT (*2, *3A, *3B, *3C) alleles are known to account for 80-95% of a decreased enzyme activity, and therefore, identifying the presence of these alleles supported by phenotypic measurement of the enzyme activity can reveal patient's TPMT status. Evaluation of the levels of thiopurine metabolites further supports the practice of appropriate dose adjustment by providing the efficient monitoring of drug cytotoxicity. CONCLUSION: We hereby review the thiopurine pharmacogenetics and the methods applied in common practice to evaluate patient's TPMT status.

Genotype-Phenotype Correlations of ß-Thalassemia Mutations in an Azerbaijani Population.

ß-Thalassemia is the most common inherited disorder in Azerbaijan. The aim of our study was to reveal genotype-to-phenotype correlations of the most common ß-thalassemia mutations in an Azerbaijani population. Patients with codon 8 (-AA), IVS-I-6 (T>C), and IVS-II-1 (G>A) mutations, which are reportedly the most common ß-globin gene mutations among the local population, were tested for hematologic parameters. Fifty-five previously tested patients with known genotypes were included in the study. Hematologic indices and hemoglobin fractions were tested in order to reveal the phenotypic manifestation of the mutations. The results obtained indicate that clinical presentation varies between different ß-globin gene mutations: individuals with IVS-I-6 (T>C) mutations showed milder presentation than those with codon 8 (-AA) and IVS-II-1 (G>A), which is associated with the molecular basis of the mutations. These data can be of assistance to predict clinical presentation and select the best possible therapeutic approach via early genotype identification.