[Particular features of erythropoiesis in high altitude and possibilities of applying of hypoxic hypoxia methodology for the patients with hemopoietic suppression].

Conditions of hypoxic hypoxia at 3200 m height exert significant positive changes in hemopoiesis, normalizing erythropoiesis and coagulation system. Hypoxic climate therapy can be regarded as an additional efficient method to the pathogenetic treatment for patient with unpainful aplastic anemia and idiopathic thrombocytopenic purpura. It should be emphasized that patients must be out of immunosuppressive therapy when getting high altitude stationary.

[Dynamic beta-endorphin determination in hematologic patients].

AIM: To perform a dynamic study of beta-endorphin, hypoxia-inducible factor-1alpha (HIF-1alpha), and cytokines in hematologic patients. SUBJECTS AND METHODS: Fifty-nine patients with different types of acute leukemia (AL), 30 with anaplastic anemia (AA), 24 with thrombocytopenic purpura, and 20 healthy volunteers were examined during their 40-day stay at 3200 m above sea level. beta-Endorphin and HIF-la were measured by a sandwich-type enzyme immunoassay using the Abcam antibodies. Cytokines (interleukin (IL)-2, IL-6, and tumor necrosis factor-alpha) were estimated by enzyme immunoassay applying the Pro Con kits (Saint Petersburg). RESULTS: Serum beta-endorphin concentrations were 1.5-2-fold above the normal values in the majority of patients with AL. The patients with initial leukocytosis at onset of disease were noted to have elevated white blood cell beta-endorphin concentrations up to 85.9 +/- 22.4 pg/ml; moreover, during chemotherapy this index increased about two times (170.74 +/- 33.8 pg/ml). There was a direct correlation between the concentrations of beta-endorphin and HIF-1alpha (r = 0.9) and an inverse correlation between the levels of IL-6 and beta-endorphin (r = -0.7). On ascending to 3200 m, under the conditions of hypoxic hypoxia the patients with AA or idiopathic thrombocytopenic purpura showed a considerable increase in serum beta-endorphin concentrations, mainly in the acute period of being at high altitudes. CONCLUSION: Stress factors (tumor, use of cytostatics, pain, anemia, hypoxia, high environment temperature) stimulate the elaboration of beta-endorphin, particularly in the white blood cells of patients with AL during chemotherapy. The highest elevation in the index was seen during acute adaptation to hypoxic hypoxia.

[Determinants for iron overload in patients with acute leukemias and aplastic anemia].

AIM: to reveal the determinants of the development of iron overload in patients with acute leukemias (AL) and aplastic anemia (AA). SUBJECTS AND METHODS: The investigation included 104 patients, including 64 with various types of AL, 31 with AA, and 9 with myelodysplastic syndromes (MDS). A group affiliation and an erythrocyte phenotype were determined from rhesus system antigens in all the patients and the HFE gene was studied to identify mutations. For control of siderosis, the authors determined serum iron (SI) by a colorimetric technique, by applying the kits of the AGAT firm (Russia), serum ferritin (SF) by an immunoradiometric method, by using the kits of Immunotech (Czechia). The volume of transfusion was estimated in the period of June 2007 to November 2009. RESULTS: There is evidence for a relationship between the higher level of SF and the number of transfusions. SF was 1046.1 microg/l in patients, H63D heterozygous carriers who had received less than 10 packed red blood cell transfusions and 2856 microg/l in those who had 20 transfusions (p < 0.005). HFE gene mutation carriage accelerates iron accumulation and is an additional risk factor for siderosis. In patients with transfusion chimeras and a rare phenotype in terms of rhesus antigens, packed red blood cell transfusion results in a much more increase in iron stores. CONCLUSION: The most important factor of iron overload acceleration is no specific choice of packed red blood cells for patients with rare combinations of red blood cell antigens and for those with artificially induced chimeras.

[Regulatory mechanisms of iron metabolism in patients with acute leukemia].

AIM: To estimate the levels of hepsidine, HIF-1alpha, erythropoietin, other proteins of iron metabolism; to characterize dysregulation of metabolic processes in leukogenesis. MATERIAL AND METHODS: Thirty eight patients with newly diagnosed acute leukemia (AL) were divided into three groups by anemia severity: group 1 (Hb > 90 g/l), group 2 (Hb 90-70 g/l), group 3 (Hb < 70 g/l). Erythropoietin concentration was measured with enzyme immunoassay, serum ferritin (SF)--by radioimmunoassay; HIF-1alpha, hepsidine--by sandvich enzyme immunoassay with use of monospecific antisera and monoclonal antibodies against relevant antigens. RESULTS: In AL patients SF before treatment was 10 times higher than in healthy subjects, administration of cytostatics elevated this concentration even more. Hepsidine and HIF-1alpha are also elevated. Treatment reduces hepsidine level twice in all the groups. This may be due to reduction of the tumor mass. Erythropoietin was 20-35 times higher in all the patients, especially in myelotoxic agranulocytosis (up to 1000 mU/ml) with reduction after recovery of hemopoiesis (in some patients to normal values 20-30 mU/ml). Hepsidine and HIF-1alpha concentrations were also maximal in myelotoxic agranulocytosis (20-28 pg/ml). After recovery of hemopoiesis these values fell to initial values 7-9 pg/ ml). Transfusion of donor erythrocytic mass normalized HIF-1alpha concentration and decreased that of hepsidine. Its elevation and high HIF-1alpha were observed after the transfusion in 17% patients. CONCLUSION: Disorders in regulatory mechanisms in AL patients throughout the observation confirm the role of the proteins studied in homeostasis. Changes in HIF-1alpha and hepsidine concentrations can be used as indicators of transfusion efficacy.

[Value of dynamic study of cytokines in serum and leukocytes in patients with acute leukemia].

A number of interleukins and iron metabolic parameters were studied in acute leukemia over time. Regulation of hemopoiesis and the cytokine network has been found to be impaired, which appears as the increased synthesis of ferritin and hepsidin by macrophages that are activated by imbalance in the cytokine network. A severe impairment of the parameters responsible for a regulatory process is shown to occur in leukemia. There is no complete normalization of these parameters during hematological remission, which is likely to lead to a further relapse of leukemic process.

[Hyperhomocysteinemia--one of the factors underlying thrombotic complications in patients with chronic myeloproliferative diseases].

AIM: To assess incidence of hyperhomocysteinemia (HHC) in patients with chronic myeloproliferative diseases (CMPD) and to analyse possible correlation between an elevated concentration of plasma homocystein (HC) and thrombotic complications. MATERIAL AND METHODS: The trial enrolled 61 patients: 39 CMPD patients with thrombotic complications and free of them, 22 nonhematological patients with thrombosis. The control group consisted of 40 healthy donors. The examination protocol included determination with standard methods of HC plasma concentration, platelet and plasma components of hemostasis, mutation of factor V Leiden gene, prothrombin and methylenetetrahydrofolate reductase (MTHFR). RESULTS: Mean HC concentration in the serum in CMPD patients was 19 +/- 1.7 mcmol/l which appeared higher than in healthy donors (12 +/- 1.3 mcmol/l). The highest HC was in patients with subleukemic myelosis (SLM)--23 +/- 2.3 mcmol). No difference in HC concentration in plasma was observed in CMPD carriers of homo- or heteroxygous mutation of C667T gene or CMPD patients without the mutation. In CMPD content of factor VIII was higher in HHC than in normal HC (222 +/- 26.5 and 116 +/- 20%, respectively, p = 0.002). For von Willebrand factor 202 +/- 15.6 and 120 +/- 14.6%, respectively (p < 0.003). HC reduction in response to vitamin therapy was the greater the higher its initial level was. CONCLUSION: There is correlation between HHC and thrombosis in CMPD patients. HC concentration may depend on the proliferative stage of CMPD. As HC is a significant independent factor of thrombotic complications risk, it is necessary to detect and treat HHC.

[Chronic myeloproliferative diseases and pregnancy].

AIM: To analyse the course of pregnancy in chronic myeloproliferative diseases (CMPD) with hyperthrombocytosis, primarily, essential thrombocytemia. MATERIAL AND METHODS: The analysis of thrombogenic risk factors covered literature data and 8 cases observed by the authors. RESULTS: Six pregnant women received long-term treatment with preparations of interferon-alpha in a dose 9-20 million IU a week (both before and during pregnancy). Rapid reduction of hyperthrombocytosis (1100-4000 x 10(9) l) and the absence of a negative effect on development of the fetus were seen in all the cases. Normal delivery on week 37-39 was in 4 patients, spontaneous abortion on week 24 was provoked by a car accident. Three gravidas (gestational week 28, 33 and 34) are still under observation. Lupus anticoagulant or elevation of anticardiolipin antibodies level was detected in 4 of 8 patients, 2 patients had heterozygous mutation of methylentetrahydrofolatereductase genes and factor V (Leiden). These patients were given lannacher, faxiparine, folic acid and discrete plasmapheresis (in 2 cases). CONCLUSION: Gravidas with hyperthrombocytosis, if not contraindicated, must be treated with aspirin and interferon-alpha preparations at any gestational term. Moreover, it is necessary to exclude additional most prevalent causes of thrombophilia for adequate prevention of thromboses.

[Characteristics of erythropoiesis regulation in population living at high altitude]. / Osobennosti reguliatsii éritropoéza u zhitelei vysokogor'ia.

AIM: To study characteristics of iron deficiency anemia (IDA) in native population of high altitude territories. MATERIAL AND METHODS: 1300 women living in Tien-Shan areas were examined for serum and erythrocytic ferritins (SF and EF), general iron-binding capacity of the serum, transferrin saturation with iron, transferrin, soluble transferrin receptor (TR), erythropoietin (EP), Hb, erythrocytes, erythrocytic indices. RESULTS: Regulation of erythropoiesis in women living at high altitude is specific, i.e. disagreement between complete depletion of iron (by SF and EF) and normal level of Hb, erythrocytes, EP and TR. CONCLUSION: It is suggested that in population living at high altitudes (3000 m above the sea level and higher) long-term adaptation to hypoxia gave rise to an original mechanism of erythropoiesis regulation when all coming iron participates in hemoglobin synthesis without iron deposition. Under hypoxic hypoxia regulation of erythropoiesis is directed to prevention of tissue hypoxia.

[Clinical significance of determining the soluble transferrin receptor]. / Klinicheskoe znachenie opredeleniia rastvorimogo transferrinovogo retseptora.

The aim of the study was to work out a method for determining the soluble transferrin receptor (TfR) on the basis of the direct immune-enzyme analysis, including its dynamic testing in patients with anemia of various etiologies. Reagents, needed for the above analysis, i.e. TfRs and antibodies to them, were obtained to implement the set goal. TfR was isolated by Kanevsky's affine chromatography from the homogenate of patients. The thus isolated TfR was used to immunize and to obtain the monoclonal antibodies. The conjugate of horseradish peroxidase (HP) and antibodies to TfR were made use of to determine the quantity of bound antibodies. One type of antibodies were immobilized in plates of the "Nunk" company (Denmark), and the other type of monoclonal antibodies were conjugated with HP. A reliably higher TfR in iron-deficiency anemia was shown during the determination of the TfR level in 36 donors and 266 patients. After a conducted ferrotherapy, the TfR level went down approaching the normal value. The TfR level was related with a disease stage and activity of the prolipherative process in cases of autoimmune hemolytic anemia, B12-folate-dependent anemia, lympho- and myeloprolipherative diseases and in oncology patients. The elaborated method opens up new possibilities for the differential diagnosis of anemia and provide for objective criteria of a conducted therapy.