CLINICAL APPLICATION OF RECOMBINANT ERYTHROPOIETIN IN BETA-THALASSAEMIA INTERMEDIA.

Research objective is to study the efficacy of recombinant erythropoietin (epoetin alfa) as alternative method of treatment beta-thalassemia intermedia. Study involved 58 patients with beta-thalassemia intermedia (23 women and 35 men). In all observed patients was defined levels of hemoglobin (Hb), red blood cells (RBC), erythrocyte indexes (MCV, MCH, MCHC), hemoglobin fractions (HbA, HbA2, HbF), serum ferritin, serum erythropoietin before and after administrated rEPO. All patients received rEPO during 6 month at the dose - 10000 IU subcutaneously. The majority of patients - 39 (67%) had a good response to rEPO (increase in hemoglobin level more than 20 g/l); 16 patients (28%) had a mean response (increase in Hb 10 - 20 g/l); in 3 (5%) patients occurred poor response to rEPO therapy (increase in Hb <10 g/l). After rEPO treatment of beta-thalassemia intermedia patients there was a statistically significant change in the number of RBC, levels of HbF and sEPO. The evaluation of interdependence between the indices of the baseline sEPO and increased Hb values in patients after rEPO treatment revealed the presence of the reverse direct relationship (r=-0.67). Based on the results, it can be concluded that the use of rEPO in complex therapy of beta-thalassemia intermedia leads to increased levels of Hb and consequently reducing the need for blood transfusions, and accordingly expected to prevent severe complications of blood transfusion (alloimmunization, hypersplenism, iron overload, contamination transmissible infections) facilitating normal growth and development, and a better quality of life.

[THE PHENOTYPIC MANIFESTATIONS OF THALASSEMIC MUTATIONS IN AZERBAIJAN].

The aim of this study was to investigate the phenotypic expression of the most common ß-globin gene mutations - Codon 8 (-AA), IVS 1.6 and IVS 2.1 in heterozygous, homozygous and compound with HbS in Azerbaijan. In total, 137 homozygous and heterozygous ß-thalassemia and S/ß-thalassemia individuals were included to the study. Red blood cell count, hemoglobin, hematocrit and erythrocyte indices were studied. Furthermore, hemoglobin fractions were analyzed by electrophoresis on cellulose acetate films and high performance liquid chromatography. Thalassemic mutation were detected using reverse hybridization method via commercial kits. Increased red blood cell count, decreased erythrocyte indices and high HbA2 was observed in heterozygous individuals causing thalassemia minor. The results obtained during the screening of mutations and analysis of hematological parameters clearly showed that the phenotypic expression of homozygous mutations, on the other hand, varies between different mutations.

[The comparative characteristic of two methods of detection of mutations of beta-globin gene].

Nowadays, two methods of detection of mutations of beta-globin gene are applied: amplification of refractory mutation system and reverse dot-blot-hybridization. The study was implemented to comparatively analyze effectiveness of these methods of molecular diagnostic in detection of thalassemic mutations in Azerbaijan. The examined sample consisted of 82 patients with both homozygous and heterozygous thalassemia and drepanothalassemia as well. In examined patients 146 mutant alleles were detected; 132 were thalassemic ones (16 various mutations) and 14 ranked among hemoglobinosis S (cod6(A>T)). The comparison of effectiveness of mentioned methods made it possible to conclude that both compared methods fit the diagnostic of thalassemic mutations. However the method of inverse dot-blot-hybridization has a number of advantages and is the best choice for Azerbaijan.

Efficiency of recombinant erythropoietin administration in hemoglobinopathy H.

Alpha-thalassemia is widely spread in human population and one of the most common types of α-thalassemia is hemoglobinopathy H which develops with mild microcytic hypochromic anemia, hepatosplenomegaly and jaundice. The basic method of anemia correction is blood transfusion. However this method has crucial deficiencies. As it is known recombinant erythropoetin (rEPO) contributes to erythroid proliferation and could be used for anemia treatment. The aim of the study was to qualify efficiency of administration rEPO in complex therapy of hemaglobinopathy H. Study involved irregularly transfused 14 patients with hemoglobinopathy H (2 males and 12 females). Control group included 30 healthy persons. Recombinant erythropoietin (Eprex) administrated hypodermically 10,000 units 3 times a week during 6 months. Average hemoglobin level before treatment was 62 g/l. Responses to the rEPO treatment varied from 9 to 70 g/l, 9 (64%) of patients had a good response, showed an increase in hemoglobin level more than 20 g/l. In 4 patients (29%) had a moderate response (10-20 g/l), but only in 1 (7%) patient occurred poor response. Changing the parameters of erithrocyte indices, hemoglobin fractions, serum iron and serum ferritin level are not statistically significant. It can be concluded that the use of rEPO in complex therapy of hemaglobinopathy H, leads to increased levels of hemoglobin and consequently reducing the need for blood transfusions.

[The serum erythropoietin under intermediate B-thalassemia].

The study explored the level of serum erythropoietin in patients with intermediate beta-thalassemia to determine possible correlations with hemoglobin level and fetal hemoglobin level and other parameters. The sampling consisted of 58 examined patients with intermediate beta-thalassemia. The contrl group consisted of 30 healthy persons. All patients underwent the identification of erythrocytes count, hemoglobin level, hematocrit indicator, erythrocyte indices (MCV, MCH, MCHC), fetal hemoglobin level, hemoglobin A2 level. Also, to all patients the identification of serum erythropoietin and ferritin levels was applied The presence/absence of relationship between these indicators was established using the correlation coefficient calculation. The study results demonstrated that under intermediate beta-thalassemia are observed statistically valid decrease of level of hemoglobin and erythrocytes, increase of percentage of fetal hemoglobin, hemoglobin A2 ferritin and serum erythropoietin as compared with standard values. The analysis of correlation relationships between indicators under study revealed that under the intermediate beta-thalassemia in many cases the failure of interdependencies of indicators marked as normal are established. The presence of clear-cut inverse interrelationship between serum erythropoietin level and hemoglobin level has to be taken into account in the process of treatment of this disease with medications of recombinant erythropoietin.

[Diagnosis of beta-thalassemia]. / Diagnostika beta-talassemii.

The purpose of the case study was to evaluate the potentialities of using the data found by hematology analyzers in the diagnosis of beta-thalassemia. Eighty (80) persons with homozygous beta-thalassemia, 180 carriers of the beta-thalassemia gene and 50 healthy subjects were examined. Counts of erythrocytes, content of hemoglobin, hematocrit and erythrocytes indices (MCV, MCH, MCHC) were determined in all of them. Besides, also in all cases, the osmotic erythrocyte resistance was investigated; the content of fetal hemoglobin was determined; electrophoresis of hemoglobin in the cellulose-acetate films was made as well as a lack or presence of the interplay between the studied parameters was established through estimating the correlation factor. The results showed significant changes in many parameters both in homo- and hetero-thalassemia versus such parameters in healthy subjects. Blood tests by hematology analyzers in cases of carrier-state of the thalassemia gene were acknowledged to be extra valuable since no clinical signs are available in such disease variation. MCV and MCH are most useful parameters in the diagnosis of heterozygous thalassemia for a reliably direct correlation was found between them.

[Heterozygote carrier state of hemoglobin S in Azerbaijan]. / Geterozigotnoe nositel'stvo gemoglobina S v Azerbaidzhane.

Clinico-hematological characteristics of 123 HbS heterozygotes have been presented. A total of 16 subjects with the level of HbS less than 30% were detected. Intraerythrocytic inclusions observed in single cells, as well as a significant decrease in the mean hemoglobin level, ESR and mean cell hemoglobin denote the presence of concomitant alpha-thalassemia-I in this group of heterozygotes. A comparative evaluation of phenotypic manifestations depending on HbS level has been presented. It is shown that shifts in the relative content of HbS lead to the widening of the borders of phenotypic manifestations.