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Objective: to establish the relationship between quantitative and qualitative parameters of peripheral blood cells(lymphocytes, neutrophilic granulocytes, monocytes, platelets) depending on the type of somatic diseases andannual internal radiation doses from 137Cs in children - residents of radiologically contaminated territories in thelate period after the Chornobyl Nuclear Power Plant (ChNPP) accident., Materials and Methods: There were 175 children included in the study comprising residents of radiologically con-taminated territories (n = 79) aged from 4 to 18 years. Annual internal radiation doses in children from 137Cs rangedfrom 0.004 to 0.067 mSv. Certain blood parameters were assessed in a comparative mode in children having got theradiation doses up to 0.01 mSv and higher. The comparison group (n = 96) included children living in settlementsnot attributed to the radiologically contaminated ones. Incidence and type of somatic diseases and its impact onquantitative and qualitative changes in blood parameters (i.e. lymphocyte, neutrophilic granulocyte, monocyte, andplatelet count) were studied. The cell size, state of nucleus, membranes and cytoplasm, signs of proliferative anddegenerative processes were taken into account., Results: Incidence and type of somatic diseases in children did not depend on the annual internal radiation dose.Number of cases of monocytosis was significantly higher among the children exposed to ionizing radiation than inthe comparison group (16.6 % vs. 7.3 %). There were, however, no correlation between these changes and radiationdoses. Number of activated blood monocytes with cytoplasmic basophilia and residues of nucleoli in nuclei washigher in individuals with internal radiation doses > 0.01 mSv. A direct correlation between the qualitative param-eters of monocytes and internal radiation doses was established (rs = 0.60; р < 0.001), as well as a direct correlationof different strength between qualitative parameters of blood cells, indicating their unidirectional pattern depend-ing on the somatic morbid conditions. Regardless of annual internal radiation dose, there was an increase in thenumber of degenerative and aberrant cells vs. the comparison group (р < 0.05), which could be due to the role ofnon-radiation factors., Conclusions: Results of the assessment of quantitative and qualitative parameters of peripheral blood cells reflect-ed the state of morbid conditions in children and are of a diagnostic value. The identified dose-dependent changesin monocyte lineage of hematopoiesis may be the markers of impact of long-term radionuclide incorporation withfood in children living in environmentally unfavorable conditions after the ChNPP accident., (V. G. Bebeshko, K. M. Bruslova, L. O. Lyashenko, T. I. Pushkariova, N. M. Tsvetkova, S. G. Galkina, V. V. Vasylenko, Zh. S. Yaroshenko, A. L. Zaitseva, L. O. Gonchar, S. M. Yatsemirskyi.)

Objective: to assess the thyroid disease in the late observation period in children who had received chemo- andradiotherapy for the acute lymphoblastic leukemia (ALL) taking into account gender, age period and disease sub-type., Materials and Methods: The incidence and nature of thyroid disease (hypothyroidism, thyroiditis, and thyroid can-cer) were studied in children-survivors of acute lymphoblastic leukemia (ALL) being in remission from 6 to 25 years.The distribution of patients by leukemia subtypes was as follows: «common» - 67.4 %, pre-B - 23.9 %, pro-B andT-cell - 4.3 %. Children had been receiving chemo- and radiotherapy according to the protocol. Regarding the ageof patients at the time of ALL diagnosis the prepubertal, pubertal and postpubertal periods were taken into account.The endocrine diseases in family history, body weight at birth, serum content of free thyroxine, pituitary thyroid-stimulating hormone, cortisol, iron, ferritin and thyroperoxidase antibodies were evaluated and assayed., Results: Thyroid disease in children was emerging in the first 2-3 years after the ALL treatment with an incidenceof 22.8 % (hypothyroidism - 14.1 %, autoimmune thyroiditis - 7.6 %, papillary cancer - 1.1 %). Seven children inthis group had received radiotherapy (12-18 Gy doses) on the central nervous system (CNS). No correlation wasfound between the radiation exposure event itself, radiation dose to the CNS and thyroid disease in the long-termfollow-up period. Thyroid cancer had developed in a child 11 years upon chemo- and radiotherapy. Hypothyroidismwas more often diagnosed in the patients of prepubertal age (rs = 0.49). There were endocrine diseases in thefamily history in about a half of children, being significantly higher than in the general sample (р < 0.05). The bodyweight at birth of a child who had later developed hypothyroidism was less than in children having got thyroiditis(rs = 0.57)., Conclusions: Disorders in endocrine regulation and of thyroid in particular can affect the prognosis of blood can-cer course in the long-term follow-up in children, especially in prepubertal age, which requires systematic supervi-sion by hematologist and endocrinologist., (V. G. Bebeshko, K. M. Bruslova, L. O. Lyashenko, N. M. Tsvіetkova, S. G. Galkina, Zh. S. Yaroshenko, L. O. Gonchar, O. Y. Boyarska, V. F. Kuzmenko, I. V. Trykhlib, N. V. Kavardakova.)

Objective: To determine the influence of iron metabolism on the prognosis of acute lymphoblastic (ALL) and (AML)myeloblastic leukemia at the different phases of chemotherapy in children after Chоrnobyl accident., Materials and Methods: 333 children (295 - ALL, 38 - AML) were examined at the stages of chemotherapy. Thecomparison group included 93 children without leukemia. Acute leukemia variants, patients survival, relapses, thenature of disease (live child or died), iron methabolism (morphometric parameters of erythrocytes, SI, SF, STf, TS),manifestations of dyserythropoiesis, bone marrow sideroblast and patients radiation dose were taken into account., Results: In 295 patients with ALL the following variants of leukemia were established: pro-B-ALL in 23, «common»type of ALL in 224, pre-B-ALL in 29, T-ALL in 19. Thirty eight patients were diagnosed with AML (11 - M1, 19 - M2,8 - M4). Doses of radiation in patients with AL were (2.78 ± 0.10) mSv and they did not correlate with clinical andhematological parameters, disease variant. Relapse rates and shorter survival were in patients with T-ALL, pro-B-ALLand AML with SF levels > 500 ng/ml (p < 0.05). The amount of children with normochromic-normocytic anemias andmanifestations of dysplasia of erythroid lineage elements was greater in the AML than in ALL. SF content in patientswas elevated during chemotherapy and was lower than the initial one only in the remission period. Transferrin wasreliably overloaded with iron: TS (70.2 ± 2.3) % compared with the control group (32.7 ± 2.1) %. Correlationbetween TS and survival of patients was detected (rs = -0.45). Direct correlation between the number of iron granules in erythrocariocytes and SF level (rs = 0.43) was established, indicating the phenomena of ineffective erythropoiesis., Conclusions: The negative influence of iron excess in the patients body on the hemopoiesis function, manifestations of ineffective erythropoiesis and the course of acute leukemia in children have been established. Changes inferrokinetic processes in children can be the basis of leukemоgenesis development., (V. G. Bebeshko, K. M. Bruslova, L. O. Lyashenko, N. M. Tsvietkova, L. O. Gonchar, S. G. Galkina, A. L. Zaitseva, L. S. Reznikova, S. M. Iatsemyrskii, L. O. Tsvet.)

Objective: Assessment of role of the bone marrow colony-forming efficiency in plasma cell myeloma patients at different stages of treatment as a prognostic criterion for the disease course., Materials and Methods: The colony forming efficiency (CFE) was assayed in stage I-II plasma cell myeloma (PCM)patients (n = 37) aged 42-73, namely in patients survived after the Chornobyl NPP accident (n = 21) and persons notexposed to ionizing radiation (n = 16). There were 11 males exposed to ionizing radiation and having got stage I PCM,9 males and 3 females exposed and having got stage II PCM, 3 males and 3 females not exposed and having got stageI PCM, 6 males and 2 females not exposed and having got stage II PCM. Healthy persons (n = 20) were included in thecontrol group., Results: Number of the bone marrow (BM) granulocyte-macrophage colony-forming units (CFU-GM) in both exposedand not exposed PCM patients depended on a disease stage. CFU-GM was (16.7 ± 1.2) in the stage I PCM patients vs.(11.1 ± 1.1) in the stage II PCM ones both being lower (p < 0.05) compared to control (64.5 ± 2.2). Changes in cluster formation were similar, i.e. (37.7 ± 1.6) and (19.4 ± 1.3) correspondingly in the stage I and stage II PCM patients.Respective values in control were (89.8 ± 3.6). The CFE in stage I and stage II PCM patients at the time of diagnosiswas lower (5.7 ± 1.5 and 2.4 ± 1.1 respectively) vs. control (39.5 ± 1.51, p < 0.05), but has increased in remission upto (29. 6 ± 1.8) and (13.8 ± 1.2) respectively. There was no difference at that between the irradiated and non-irradiated patients. Number of the fibroblast colony-forming units (CFU-F) in the stage I and stage II PCM patients duringdiagnosis, namely (43.9 ± 5.4) and (22.5 ± 3.7), was lower (p < 0.05) vs. control (110.5 ± 4.9). Upon reaching remission the CFU-F value increased significantly (p < 0.05), reaching (87.4 ± 4.2) and (55.6 ± 2.7) correspondingly in thestage I and stage II PCM patients., Conclusion: Dependence of the BM cell CFE on the stage of PCM and presence or absence of remission was established. Prognostic value of the CFE of BM CFU-GM in terms of life span of patients was shown (Ro Spearm = 0.39,p < 0.02), namely in case of CFE > 20 before the polychemotherapy administration the life span of PCM patients wassignificantly longer vs. cases of CFE < 20., (Zh. M. Minchenko, T. F. Liubarets, V. V. Balan, O. O. Dmytrenko, T. Yu. Shlyakhtichenko, V. O. Moyseyenko, Yu. O. Silayev, V. G. Bebeshko.)

Objective: Elucidation of relationship between the levels of thyroid-stimulating hormone (TSH), free serum thyroxine, serum and urine cortisol and parameters of erythroid lineage of hematopoiesis to estimate the thyroid functionin children of prepubertal, pubertal, and postpubertal age permanently residing under a low-dose radiation exposureto determine the premorbid state of thyroid function., Materials and Methods: Children aged 3 to 18 years old (n = 203) living in the most intensively radionuclide-contaminated regions of Kyiv, Zhytomyr and Chornihiv oblasts of Ukraine after the Chornobyl NPP accident wereenrolled. Complaints of ossalgia, arthralgia, fatigue, bone fractures in the history, bone dysembryogenetic stigmata,hypermobility syndrome degree, and types of somatic diseases were taken into account. Peripheral blood countparameters, biochemical indices of blood serum were studied, namely the levels of total protein, cholesterol, creatinine and alkaline phosphatase activity. Levels of the free thyroxine, pituitary TSH, serum and daily urine cortisol, anddoses of radiation exposure were determined., Results: The radiation dose values in children ranged from (0.35 ± 0.09) mSv to (0.54 ± 0.12) mSv. There was nodifference between the parameters of erythroid lineage of hematopoiesis depending on radiation dose. At the levels of serum TSH up to 1.0 μIU/ml no correlation was found with cortisol levels; at TSH levels of 1.0-3.0 μIU/ml thecorrelation coefficient was r = 0.31; at TSH levels higher than 3.0 μIU/ml the correlation coefficient was r = 0.61probably indicating a compensatory role of adrenal cortex in children at risk of thyroid disease development. In children with joint hypermobility grade II there was a higher incidence of dentofacial anomalies (χ2 = 6.9), deformitiesof lower extremities (χ2 = 6.9), and dental caries (χ2 = 4.3) (p < 0.05). There was a direct correlation between theserum TSH level (over 3 μIU/ml) and micrognathia (brachygnathia) (r = 0.62) indicating the impact of thyroid disease on dentofacial development. The TSH at a level of upper limit of the reference range values may contribute toa decreased RBC count in peripheral blood, increased average volume and hemoglobin content in erythrocyte beingassociated with the initial manifestations of thyroid dysfunction., Conclusions: Abnormal endocrine regulation of hematopoiesis affects the connective tissue, stromal microenvironment of bone marrow, and accordingly the erythroid branch of hematopoiesis in children, which may be relevant inthe development and course of oncohematological diseases., (V. G. Bebeshko, K. M. Bruslova, O. Y. Boyarska, L.O. Lyashenko, N. M. Tsvyetkova, L. O. Gonchar, T. O. Chernysh, I. V. Trychlib, V. D. Pismenniy, Z. S. Yaroshenko, V. V. Vasylenko.)

Objective: establishing the types and frequency of disembriogenetic stigma in children with joint hypermobility given the clinical and laboratory features, genetic component and endocrine regulation of these disorders in a late period upon the accident., Materials and Methods: Children (n = 109) inhabiting the radiologically contaminated territories and having the connective tissue dysplasia (CTD) signs were involved in the study. Diseases in family history, ossalgia complaints, fractures in a personal history, bone disembriogenetic stigma, joint hypermobility, type of somatic diseases, blood serum biochemical parameters (namely calcium, alkaline phosphatase, total protein, cholesterol, creatinine, iron, ferritin content), serum cortisol, free thyroxine, pituitary thyroid-stimulating hormone (TSH) levels, free amino acid composition in urine and radiation dose were considered., Results: Radiation doses in children having the CTD ranged from (0.37 ± 0.11) mSv to (0.56 ± 0.10) mSv with no difference from that in those without CTD. Joint hypermobility (JHM) correlated with cancer in family history (rs = 0.53) and lower extremity varicose vein disease (rs = 0.40) (p < 0.05). Incidence of ossalgia, easy fatigability, and bone fractures was higher in children with CTD. Anomalies of the dentofacial system were first in line (38.5 %) in these children. Proportion of children with grade II JHM and platypodia was lower (rs = 0.42), but with lower extremity deformations was higher (rs = 0.68) (p < 0.05) vs. in the control group. Iron and ferritin deficiencies both with lymphocytosis were more common in children with CTD than in the comparison group (p < 0.05). The increased content of oxyproline, lysine, proline both with glycine deficiency were detected in children having the CTD, i.e. an imbalance of amino acids from the collagen content was observed featuring a predominance of catabolic processes over anabolic ones. There was a direct correlation between the TSH level and the JHM grade (rs = 0.49), although the values of hormone concentration in these children did not exceed the reference range (maximum values were 3.3 μIU/ml)., Conclusions: The revealed abnormalities in amino acid content, ferrokinetics, and thyroid function indices can affect the collagen formation, organic matrix structure of bone tissue and significantly deregulate the hemato- poiesis. The later can underlie the pathways of haematologic malignancy development., (V. G. Bebeshko, K. M. Bruslova, T. T. Volodina, L. O. Lyashenko, N. M. Tsvetkova, I. V. Trychlib, T. O. Chernysh, V. G. Boyarskiy, L. O. Gonchar, N. V. Kavardakova.)

Objective: Experimental study of the effect profile of bortezomib in the plasma cell myeloma (PCM) patients depend- ing on a specific phenotype carrier state and a pharmacochemical characteristics of ABO system glycoproteins., Materials and Methods: The research was conducted on the 104 PCM patients, including the Chornobyl NPP acci- dent survivors (n = 49) and 65 study subjects in the comparison group. Immunogenetic criteria for positive response to the applied treatment protocols were issued according to the duration of remission, absence of infectious com- plications, and evidence of chronic renal failure as a disease complication., Results: Possibility of glycoproteins A and B participation in the formation of human biological individuality at a level of protein-protein interaction with antineoplastic drug bortezomib, which is widely used in cancer management prac- tice, in particular in the PCM treatment is considered. The glycoprotein B was shown being a selective target for borte- zomib, slowing down the recognition and interaction of antigen B with monoclonal anti-B antibody, while the agglu- tination period lengthens at that by 66 %. Assumption that the formation of bortezomib complex with glycoprotein B provides a background for interaction with the key reaction of proteasome 26S inhibition, which to some extent con- tributes to the drug effect retardation was confirmed through the quantum-chemical calculations. Equilibrium is shift- ed toward the main reaction leading to a higher drug efficacy in patients with blood groups O (I) and A (II)., Conclusions: Since the complexation occurs predominantly in alkaline medium the administration of drugs with alkaline reaction should be restricted for at least round the clock after administration of bortezomib according to its half-life in plasma in patients with B (III) blood group and chronic renal failure., (Zh. M. Minchenko, A. D. Kustovska, S. V. Prymachenko, O. O. Dmytrenko, T. F. Liubarets, T. Yu. Shlyahtichenko, V. V. Balan, V. G. Bebeshko.)

Objective: Estimation of the bone marrow haemopoietic status depending on the reasons and duration of breaks in a standard chemotherapy (BFM-ALL protocol) to predict the course of acute lymphoblastic leukemia (ALL) in chil- dren exposed to low doses of ionizing radiation after the Chornobyl accident., Materials and Methods: The ALL patients (n = 34) were examined within 5 stages of a program chemotherapy. The clinical symptoms, hemogram and myelogram data were analyzed. The radiation dose on bone marrow, initial leuko- cyte count, variants and prognosis of ALL course were accounted. Days of the stopped chemotherapy, type and fre- quency of complications (septic processes, febrile neutropenia, toxic hepatitis, granulocytopenia degree), and the prognosis of disease course (child living status, i.e. alive or died) were estimated., Results: There were abnormal differentiation processes and high percentage of lymphoblasts (86.2 ± 3.3) % in bone marrow in the 1st acute period. Hematological remission was established in all patients on the 33rd day of chemothe- rapy. In a half of cases the haematopoietic recovery occurred by a granulocyte-monocyte type. One third of patients presenting an erythroid type of haemopoiesis died later. The inverse correlation was found between the number of myelocaryocytes and disease prognosis (rs = -0.49). Breaks in chemotherapy for various reasons were recorded. The number of patients with granulocytopenia was greater at the phase 1 and 2 of protocol I and protocol M application, coinciding with a higher incidence of complications. An inverse correlations between the prediction of ALL course and sum of days of breaks between the protocol M and phase 1 of protocol II (rs = -0.56), as well as the duration of the phase 2 of protocol II (rs = -0.62) were found. The radiation dose on bone marrow was (5.37 ± 1.23) mSv. No relationship was found between the radiation doses, ALL variants and disease course., Conclusions: Prognosis of ALL course in children depends on the type of haemopoietic recovery and reasons of breaks in a standard chemotherapy. Interaction between the haemopoiesis functioning and microenvironment and that of their regulation are the key mechanisms of above-mentioned abnormalities, which is the basis for further research., (V. G. Bebeshko, K. M. Bruslova, N. M. Tsvetkova, L. O. Lyashenko, T. I. Pushkariova, L. O. Gonchar, I. V. Tryhlib, S. M. Yatsemirskyi, Yu. M. Samson, V. G. Boyarskyi, K. V. Grischenko, V. M. Polyanska, I. V. Dmytrenko.)

This population-based ecological study analyzes the prevalence of childhood leukemia in Ukraine before and after the Chornobyl nuclear power plant accident, based on the contamination status of the territory, time period, gender, and age. Three regions-Zhytomyr, Kyiv (except Kyiv city), and Chernihiv were included as areas contaminated by radioactive 137 Cs from 1 to 15 Ci/km 2 with annual effective doses exceeding 1.0 mSv, and Sumy region as the control (non-contaminated) area with 137 Cs contamination less than 1 Ci/km 2 and effective doses less than 0.5 mSv per year. The integrated database of the National Research Centre for Radiation Medicine used in the present study included 1085 childhood leukemia cases. Two aggregated periods were used for analysis: 1980-1986 (pre-accident) and 1987-2000 (post-accident). ICD-9 codes for leukemia (204-208.9) were used to perform analyses according to the extent of leukemic cells maturity (acute, chronic, and maturity unspecified leukemia), leukemic cell lineage (lymphoid, myeloid and lineage unspecified leukemia) and all leukemia cases in different age subgroups (1-4, 5-9, 10-14, and 15-19 years). Standard methods of descriptive epidemiology were used to calculate the prevalence of disease and frequency ratio in regression models. A statistically significant increase in frequency ratio for acute leukemia (1.44; 95% confidence interval (CI), 1.22-1.71), myeloid leukemia (2.93; 95% CI, 1.71-5.40), cell lineage unspecified leukemia (II) (1.48; 95% CI, 1.18-1.87) and all forms of leukemia (1.59; 95% CI, 1.36-1.86) was found for the post-accident period in highly contaminated areas. The results indicate that the frequency of childhood leukemia (and of some of its types) increased in contaminated areas during the post-accident period, suggesting that radiation exposure after the Chornobyl accident might be the cause of the increase. However, further analytical studies, with individual or at least group dose estimates, are needed to confirm a link between childhood leukemia and the Chornobyl accident.

Objective: Determine of the influence of adverse environmental factors, including irradiation, on the survival ofchildren with acute leukemia in the long-term period after the Chornobyl accident (2008-2017)., Materials and Methods: Examined 74 children with acute leukemia (АL): 64 with acute lymphoblastic leukemia(ALL); 10 - acute myeloblastic leukemia (AML). The influence of negative environmental factors was assessed bythe degree of integrated pollution of the atmospheric air, surface waters and soils with pesticides, heavy metals (Pb,Cu, Ni, Cr, Mn, Zn, Fe) and cesium isotope 137Cs. These regions were ranked on a moderately polluted (1), polluted (2),very (3) and extremely (4) polluted territories. Took into account the age of children, blood test, immunophenotyp-ic of variant the acute leukemia, survival of patients and place of residence (city / village), serum ferritin level (SF)., Results: Exposure doses of children were in the range from 0.4 mSv to 35.0 mSv (average values were (4.25 ± 0.63 mSv)and did not affect the prognosis and variants of AL. 52 children lived in moderately and polluted territories (30 wereresidents of cities, 22 - villages). 22 patients were lived in very and extremely polluted territories (4 were residentsof cities, 18 - villages). Of 74 patients with ALL and AML 24 children died (32.4 %). The smallest number ofchildren, who died, were patients with «general type» В-ALL (18.7 %), most of all children with pro-В-ALL (8 out of10) and Т-ALL (3 of 4). Of the 52 patients, the inhabitants of moderately and polluted regions, 13 patients died (25%), while out of 22 patients, who lived in very and extremely polluted areas, 11 children died (50 % share)(rs = 0.39; p < 0.05). Of the 10 patients with AML, 4 children died. Most often, children died, who were residents ofvillages. Moreover, the level of SF was significantly higher in children over 6 years, the inhabitants of villages -(406.8 ± 40.6) ng/ml, compared to younger patients - (211.2 ± 32.1) ng/ml) and residents of of cities: up to 6years - (297.4 ± 52.3) ng/ml; over 6 years - (275.6 ± 29.8) ng/ml., Conclusions: The obtained data testify to the negative influence of environmental factors, including iron, and canbe the basis for understanding the mechanisms of potentiating influence of metals and their compounds on thedevelopment of malignant diseases of the blood system in children., (V. G. Bebeshko, K. M. Bruslova, N. M. Tsvietkova, L. О. Lyashenko, T. I. Pushkareva, L. O. Honchar, A. S. Sergeeva, M. Yatsemyrskii, Y. M. Samson, I. V.Trihlib, O. I. Makovey.)

Objective: Determination of serum cortisol level in the initial period of acute leukemia in children, who exposed to ion izing radiation and other factors of Chornobyl accident, depending on their age and prognosis of disease., Materials and Methods: The study involved 283 children residents of Kyiv, Zhytomyr and Chernihiv regions. There were 90 acute leukemia patients(AL) (ALL - 56, AML - 34), and 193 people of comparison group with anemia, leukemoid reactions and lymphadenopathy. We analyzed the type of comorbid somatic pathology, diseases in the genealogy, hematological parameters, cortisol levels in blood serum and irradiation doses in all children. In patients with AL expected median survival was calculated., Results: In 28.9 % of AL children the initial cortisol content was below 200 nmol/l, in 7.8 % - higher than 500 nmol/l (in the comparison group 10.4 % and 17.1 % respectively). Among AL patients with cortisol levels below 200 nmol/l were significantly less amount of persons with chronic bacterial infections and persistent viral infections (CMV, EBV) and in the genealogy of these children allergic reactions, endocrine pathology diagnosed more often compared with patients, whose hormone levels was higher than 200 nmol/l (p < 0.05). Distribution of children from control group by gradations of cortisol, age groups, defined somatic pathology and diseases in genealogy had no difference. It is shown, that lower initial blood serum cortisol level in ALL children correlates to a greater probability of relapse (Rs = -0,67). In patients with AML a direct correlation between cortisol level and median survival was detected (Rs = 0,79). Children radiation doses were ranging from 0.08 mSv to 14.9 mSv, and there were slightly higher among residents of Zhytomyr region (8.4 ± 1.2 mSv) compared to other regions. However, these doses did not affect blood serum cortisol levels in children and the course of AL., Conclusions: These data suggest the need for correction and individualization of corticosteroid doses for optimization of AL patients treatment. Children, who have lower than normative serum cortisol levels are at increased risk of hema tologic pathology and they need for hematologic monitoring., (V. G. Bebeshko, K. M. Bruslova, T. I. Pushkareva, N. M. Tsvietkova, L. O. Lyashenko, A. S. Sergeeva, V. F. Kuzmenko, S. M. Iatsemyrskiy, Yu. M. Samson, V. G. Boyarsky, I. V. Tryhlіb.)

Objective: Evaluation of proliferation and differentiation processes of progenitor cells in bone marrow by the com position of elements of erythroid, granulocyte and platelet branches of hematopoiesis on the treatment stages in children with acute lymphoblastic leukemia (ALL), who were exposed to radiation from the Chornobyl NPP accident., Materials and Methods: The 46 children with ALL were studied, who lived in Kyiv, Zhytomyr and Chernihiv regions. Studies were conducted before the start of chemotherapy (ChT), on the 33 day of ChT (phase I), and after the com pletion of ChT (phase II). Exposure doses of patients, hemogram and myelogram parameters both with indices of mat uration of progenitor cells were evaluated. Signs of dysplasia of hematopoietic branch elements were revewed., Results: The 46 patients were studied. They have had the B ALL, namely pro B ALL (n=5), «common type» (n=36), pre B ALL (n=3), and T ALL in 2 other cases. In a debut of ALL the bone marrow was represented by lymphoblasts. Along with ChT conduction the bone marrow hematopoiesis recovered by such types, as erythroid, granulocyte, gran ulocyte whith monocytes, and uniform, when the cells number of all branches was within a normal quantity. At the phase ІІ of ChT the number of patients with hematopoiesis recovery by erythroid type decreased and number of chil dren with activation of granulocyte branch of hematopoiesis increased. In children with pro B ALL the number of erythroid elements was higher than normative at both ChT phases. A direct correlation was established between the number of myelokaryocytes (Mkc) and megakaryocytes (Mgkc) in both phase І and phase ІІ of treatment (Rs = +0.72; Rs = +0.56, respectively). There was no correlation between the radiation dose in patients (3.73 ± 0.12 mSv) and studied parameters., Conclusions: Types of bone marrow recovery were established in ALL patients after the ChT indicating to the differ ent kinetic pathways of hematopoietic progenitor cells. Evaluation of reasons of prevalence of some hematopoietic branches will allow to reveal their role in leukemogenesis and to correct the treatment programs., (V. G. Bebeshko, K. M. Bruslova, T. I. Pushkareva, N. M. Tsvyetkova, L. O. Lyashenko, O. Ye. Kuznyetsova, V. F. Kuzmenko, L. O. Gonchar, S. M. Yaatsemyrskyy.)

Objective: Identification of impact of radiation and non radiation environmental factors on development of hematopoi etic abnormalities in children and justification of criteria for the increased risk groups of hematologic diseases., Materials and Methods: The results of clinical and hematological survey of 1465 children living in Kyiv, Zhytomyr and Chernihiv regions for the period from 2008 to 2014 were presented. There were 777 children with anemia, 466 with changes in hemogram, 191 with acute leukemia. The irradiation doses, correlation of integrated pollution degree of territories with hematopoietic parameters and course hematologic diseases were estimated. Metal con tent in hair, nails, and blood was determined in 121 children., Results: We have found the most common cause of anemia in children and peculiarities of acute leukemia depend ing on the area integrated contamination. Number of children living in contaminated areas with pro B ALL and T ALL having an initial leukocytosis and unfavorable course of the disease was higher compared to the number of patients from moderately polluted regions (r = 0.47). There is a direct correlation between percentage of children with monocytosis and degree of territory contamination: the 20.2 % of such children lived in the intensively polluted areas and 10,3 % in moderately contaminated ones (p < 0.05). Content of Pb, Cu, Cr, Mn, Zn in biological tissues did not exceed the permissible MAC. Level of Zn in all children was at the lower normal threshold. Individual levels of metals in hair, nails and blood did not depend on gender of patients and their radiation doses. Criteria of increased risk of hematologic diseases in children living in ecologically unfavorable regions were grounded., Conclusions: Radiation and non radiation environmental factors influencing the development of changes in hematopoiesis and characteristics of the blood system diseases in children play a role in leukeima development processes. These findings are the basis for a further research in the field of radiobiology and ecology., (V. G. Bebeshko, K. M. Bruslova, V. V. Stankevych, N. M. Tsvietkova, L. О. Lyashenko, S. G. Galkina, T. I. Pushkaryova, V. I. Kolos, O. Ye. Kuznetsova, L. O. Honchar, S. M. Yatsemyrskii, Y. M. Samson.)

Objective: To investigate the aminoacid composition in the stromal fibroblasts of bone marrow and indexes of hemogram in children with acute lymphoblastic leukemias, which exposed to radiation from the Chornobyl accident, for evaluation of the median survival., Materials and Methods: In the study 54 patients with acute lymphoblastic leukemia, residents of Kyiv, Zhytomyr and Chernihiv regions of Ukraine were involved. Effectiveness of stromal fibroblasts colony forming units of bone marrow and aminoacid composition of fibroblasts were studied. Bone marrow examination was conducted on day33 and after stop of chemotherapy, the median survival of patients and exposure dose were evaluated., Results: In children with acute leukemias effectiveness of stromal fibroblasts of bone marrow was 2-2,3 times lower than normative. Correlation (rs = +0,83) between the number of erythroid progenitor cells in the bone marrow and proline content in stromal fibroblasts established . The correlation between patients median survival and number of granulocytes in myelogram (rs = + 0,82) was proven. Mean exposure doses in patients was (5,86 ± 1,11) mSv and it did not correlate with myelogram results, effectiveness of stromal fibroblasts of bone marrow and median survival., Conclusions: The median survival of patients with acute leukemias depends on the content of amino acids in stro mal fibroblasts of bone marrow and recover of hematopoiesis after chemotherapy., (V. G. Bebeshko, K. M. Bruslova, N. M. Tsvyetkova, T. T. Volodina, T. I. Pushkarova, L. O. Lyashenko, L. M. Panchenko, S. M. Iatsemirskii.)

Age and sexual indexies of densitometry at patients with acute leukemia (AL) and healthy children are presented. 31% of children with AL during the initial period of disease had manifestations of the osteopenic syndrome. At patients with AL more often than at healthy children anomalies of development of front part of skull are defined. The partial contribution of free and peptides-connencted oxyproline in urine at AL patients differs in comparison with control group that is caused by modification or deficiency of the corresponding enzymes. 30% of patients with AL had raised concentration of free oxyproline in urine, and lowered glycine concentration that testifies to the increased disintegration of collagen and deficiency of tile plastic material necessary for collagene-forming processes. The obtained data should be considered for forming of risk group on oncohematological pathology at children.

Diffuse large B-cell lymphoma (DLBCL) is the most common type of lymphoma, including approximately 30-40% of all B-cell non-Hodgkin lymphomas (B-NHL). Chromosomal translocations are the hallmark of genetic aberrations in B-lymphoma and are often associated with a specific subtype of B-NHL. MYC gene dysregulation due to chromosomal translocations is characteristic for the most cases of Burkitt's lymphoma. Objective. The goal of this study was to improve the diagnostic accuracy of DLBCL. Identification of chromosome 8 and 14 abnormalities including the translocation of MYC gene t(8; 14)(q24; q32) in substrate cells of lymph nodes was applied using the method of tri-color interphase fluorescence in situ hybridization (I-FISH). Materials and methods. Lymph node biopsy specimens of 17 patients with diffuse large B-cell lymphoma and three patients with Burkitt's lymphoma (including one participant of liquidation of consequences of the catastrophe at the Chornobyl NPP) were studied. The age of patients ranged from 10 to 66 years old (41.3 ± 3.7 average). Biopsy specimens fixed in paraffin. I-FISH-analysis was performed using the commercial test Vysis IGH/MYC, CEP 8 tri-color, dual fusion translocation probe (Abbott Molecular, USA). Results and conclusions. MYC gene and immunoglobulin heavy chain (IGH) gene translocations were found in four out of twenty persons. Consequently the I-FISH method allows identification of of MYC and IGH gene rearrangements in tissue cells substrate of lymphoma fixed in paraffin. Using this method the molecular-cytogenetic abnormalities were found in eight of twenty patients with B-cell lymphoma providing verification of the lymphoma diagnosis, prediction of their clinical course and advance in management i.e increase the effectiveness of therapy, in refractory lymphoma cases among others., (J. А. Misharina, V. V. Sitko, S. V. Klymenko, J. А. Minchenko, А. І. Kurchenko, Y. О. Silaev, L. O. Lyashenko, V. М. Polyanska, V. G. Bebeshko.)

Objective. To justify and evaluate the effectiveness of treatment and preventive measures taken to correct the bio-chemical and biophysical changes in the structure of bone in children exposed to ionizing radiation due to Chornobyl accident in the dynamics of leukemic process. Materials and methods. The study involved 144 patients with acute lymphoid and myeloid leukemia. Serum calcium, total protein, alkaline phosphatase, cortisol and thyroid-stimulating hormone were investigated, as well as calcium and phosphate excretion, amino acids composition in the urine, acid-base balance and buffer systems of blood, bone density. Results. A reduction of bone densitometry, calcium and cortisol levels and increase of serum alkaline phosphatase and urine oxyproline were found in children with acute leukemia in acute I phase and during the chemotherapy. In the period of remission the majority of patients did not differ from the comparison group. The signs of osteopenic syndrome that require monitoring of the bone structure status were revealed in 10.0 % of children of the comparison group. Radiation doses of leukemia patients were 3.66 ± 0.11 mSv. No relationship between radiation dose and indicators that reflect the state of the bone tissue structures has been established. The use of complex health care measures in children with acute leukemia in remission, contributed to the positive effect of 62.0 %. Worser effect was observed in patients with an unfavorable course of the disease. Conclusions. Results of the study of bone tissue metabolism prove fore a change in the organic and mineral matrix in patients with acute leukemia exposed to ionizing radiation as a result of the Chonobyl NPP accident, and persons without hematologic diseases, and the need for treatment and prevention., (V. G. Bebeshko, K. M. Bruslova, N. M. Tsvietkova, T. T. Volodina, O. V. Kopylova, T. O. Belingio, S. A. Berezovski, M. P. Petrunko, V. G. Boyarskiy, V. M. Polianska, T. I.Pushkareva, L.P. Mishchenko.)

Objective: To establish the sources of lymphocytosis in children living in ecologically unfavorable conditions after the Chornobyl Nuclear Power Plant accident aiming the high risk group formation for oncohematological diseases among the children's population., Materials and Methods: The 226 children with lymphocytosis were surveyed. The of child's life history and kind of somatic morbidity were considered. Quantitative and qualitative parameters of blood cells, biochemical indices including immunoglobulins (A, M, G), circulating immune complexes, phagocytosis indices were assayed., Results: Children with lymphocytosis were more often born with a high bodyweight, they demonstrated manifestations of constitutional lymphatism, lymphadenopathy and more often had respiratory infections. Quantitative and qualitative changes of hemopoietic elements depended on a type of a disease. Activation of lipid peroxidation, dysimmunoglobulinemia and increased level of circulating immune complexes were revealed. Treatment and preventive actions promoted normalization of hemogram indices in 58% of children surveyed., Conclusions: The abnormalities in immune indices, activated lipid peroxidation in membranes of hemopoietic elements, and manifestations of granulocytopoietic disorders were established it children with lymphocytosis. Therefore these children can be attributed tot the group of increased risk on oncohematological disease., (Bebeshko V. G., Bruslova K. M., Chumak A. A., Cvjetkova N. M., Kuznjecova O. Je., Gonchar L. O., Parhomenko V. M., Pushkarova T. I., Bojarskyj V. G., Pleskach O. Ja., Abramova T. Ja., 2013.)

Unlabelled: The objective was to study the immunogenetic component contribution to the formation of post-radiation effects in the long-term period after radiation exposure at the level of the human immune response as a prognostic criterion for risk assessment of radiation-associated somatic diseases. Study object was the convalescents of acute radiation syndrome (ARS) of the first grade of severity, 88 patients with a similar radiation history but with unconfirmed ARS, 73 patients being the liquidators of the Chornobyl accident consequences (LCAC) with chronic ischemic heart disease (HIHD), 65 patients LCAC without HIHD, 120 non-exposed patients with HIHD, 256 patients with oncohematological diseases and 500 healthy people - a population control., Results: Markers of risk of realization of genetic predisposition to oncohematological and cardiovascular disease in these groups were identified on the basis of study of immunological, hematological and molecular genetic disorders in relation to immunogenetic factors., Conclusion: These data indicate that realization of HLA-genetic predisposition to the disease is one of the radiation associated multifactorial pathology pathways, and presence of radiosensitivity markers in pheno/genotype enhances the realization risk of pathological process under irradiation., (Minchenko Zh. M., Djagil I. S., Dmytrenko O. O., Dmytrenko I. V., Shljahtichenko T. Ju., Gavrylenko T. I., Bilyj D. O., Homenko V. I., Bebeshko V. G., 2013.)

Objective: To study the serum ferritin levels in children with acute leukemia, depending on the number of transfusions of RBC concentrate and period of disease., Materials and Methods: We studied the red blood count, serum iron and ferritin levels in 54 patients with acute leukemia before chemotherapy, at the time of a standardized treatment protocol, and after transfusions of RBC concentrates., Results: In the debute of acute leukemia just before treatment lauch the serum ferritin in 81.5% of children was 2.3-2.5 higher than normal. The need for transfusion of RBC concentrates was higher under serum ferritin level exceeding 500 ng/mL. The association was established between ferritin content and age of the children, variant of acute leukemia and period of the disease., Conclusions: The level of serum ferritin can be used as a marker of ferrokinetic status for timely diagnosis of iron overload in children with acute leukemias and for application of treatment-and-prophylactic actions., (Bebeshko V. G., Bruslova K. M., Cvjetkova N. M., Jacemyrskyj S. M., Pushkarova T. I., Gonchar L. O., Krukovska V. V., Zelinska A. V., Mishhenko L. P., 2013.)

Purpose: To present previously unavailable data on the use of stem cell administration to aid recovery of victims of the Chernobyl disaster. On 26 April 1986, an accident at Unit 4 of the Chernobyl Nuclear Power Plant took place during the planned test of one of the safety systems. The diagnosis of acute radiation syndrome (ARS) was confirmed in 134 individuals exposed to high levels of radiation. There were nine patients heretofore unreported in the scientific literature who underwent intraosseous injections of allogeneic bone marrow cells in Kyiv., Conclusions: Transplantation was associated with significantly shortened time to recovery of granulocyte and platelet counts in these patients. While current guidelines would certainly include the use of cytokines, these data provide an indication of the effectiveness of stem cell transplant to treat victims of radiation exposure.

Acute myeloid leukemia (AML) can develop as a secondary malignancy following radiotherapy, but also following low-dose environmental or occupational radiation exposure. Therapy-related AML frequently carries deletions of chromosome 5q and/or 7, but for low-dose exposure associated AML this has not been described. For the present study we performed genome-wide screens for loss-of-heterozygosity (LOH) in a set of 19 AML cases that developed after radiation-exposure following the Chernobyl accident. Using Affymetrix SNP arrays we found large regions of LOH in 16 of the cases. Eight cases (42%) demonstrated LOH at 5q and/or 7, which is a known marker of complex karyotypic changes and poor prognosis. In accordance with literature data, the overall survival for these patients was significantly shorter as compared to patients without this alteration (P=0,014). We could show here for the first time that exposure to low-dose ionizing radiation induces AML with molecular alterations similar to those seen in therapy-related cases.

After the Chernobyl accident, the Research Center for Radiation Medicine (RCRM) was established in Kiev (Ukraine). Its main task was to maintain a high level of emergency preparedness and be ready to examine and treat patients who suffer as a result of hypothetical radiation accident. Based on the previous experience, this institution's specialists worked out new diagnostic criteria and drug treatment schemata for acute radiation sickness, created a database on 75 patients with this diagnosis and improved educational programmes for medical students and physicians working in the field of radiation medicine. RCRM collaborates fruitfully with western partners through the joint research projects and connects with the World Health Organization's Radiation Emergency Medical Preparedness and Assistance Network centre. Collaboration with Kiev Center for Bone Marrow Transplantation allows RCRM to use aseptic wards having highly filtered air for the treatment of most severely irradiated patients.

Leukemia is one of the cancers most susceptible to induction by ionizing radiation, but the effects of lower doses delivered over time have not been quantified adequately. After the Chornobyl (Chernobyl) accident in Ukraine in April 1986, several hundred thousand workers who were involved in cleaning up the site and its surroundings received fractionated exposure, primarily from external gamma radiation. To increase our understanding of the role of protracted low-dose radiation exposure in the etiology of leukemia, we conducted a nested case-control study of leukemia in a cohort of cleanup workers identified from the Chornobyl State Registry of Ukraine. The analysis is based on 71 cases of histologically confirmed leukemia diagnosed in 1986-2000 and 501 age- and residence-matched controls selected from the same cohort. Study subjects or their proxies were interviewed about their cleanup activities and other relevant factors. Individual bone marrow radiation doses were estimated by the RADRUE dose reconstruction method (mean dose = 76.4 mGy, SD = 213.4). We used conditional logistic regression to estimate leukemia risks. The excess relative risk (ERR) of total leukemia was 3.44 per Gy [95% confidence interval (CI) 0.47-9.78, P < 0.01]. The dose response was linear and did not differ significantly by calendar period of first work in the 30-km Chornobyl zone, duration or type of work. We found a similar dose-response relationship for chronic and non-chronic lymphocytic leukemia [ERR = 4.09 per Gy (95% CI < 0-14.41) and 2.73 per Gy (95% CI < 0-13.50), respectively]. To further clarify these issues, we are extending the case-control study to ascertain cases for another 6 years (2001-2006).

The results of multidrug resistance determinants expression analysis on leukemic cells of 56 acute myeloid leukemia (AML) patients by immunophenotyping are presented. Of these, there were 21 persons exposed to ionizing radiation due to the Chemobyl accident with radiation-associated AML and 35 patients with spontaneous leukemia. The aim of this study was to determine if transport proteins (P-glycoprotein, LRP, and MDR1), apoptosis-related proteins (Fas, Bcl-2, Bax, p53, and Bcl-X(L)), and topoisomerase IIalpha expression in AML patients with the history of radiation exposure differed from those in spontaneous AML cases. Leukemic cells in patients with radiation-associated diseases compared to spontaneous AML more often overexpressed antiapoptotic oncoprotein Bcl-2 (12/21 vs. 6/35, p < 0.005) and less often demonstrated expression of Fas receptor (12/21 vs. 30/35, p < 0.05). Moreover, leukemic cells were simultaneously Fas negative and Bcl-2 positive in 4 out of 21 patients exposed to ionizing radiation but none of spontaneous cases had similar phenotype (p < 0.05). Leukemic cells in patients with radiation-associated AML compared to spontaneous cases more often were P-glycoprotein positive (12/20 vs 9/31, p < 0.05). P-glycoprotein overexpression significantly correlated with resistant disease in patients with radiation-associated AML (r = 0.47, p < 0.05), but was not a prognostic variable for the treatment outcome in terms of overall survival. Defects in pathways of drug-induced apoptosis and function of pump, that actively effluxes drugs could contribute significantly to developing drug resistance in radiation-associated AML.

The article considers the possibility of the use of current infrastructure of specialized population registers of Ukraine to study leukemia and other systemic blood diseases revealed in Chernobyl accident liquidators. Advantage and limitation of such registers in the use are discussed in the article. Ukrainian state register of people who suffered from Chernobyl accident and Ukrainian national cancer registers are the largest population registers in the country, which cover all the territory of Ukraine and contain information on each individual and may serve as source base for epidemiological studies. To solve issues on leukemia and other oncological diseases is recommended to use in complex data of specialized registers of Ukraine. It should be also taken into account necessity of using late registered cases and verifying registered diagnoses.

The results of proper investigations received under the cytogenetic examination of 225 persons (control groups, Chernobyl liquidators exposed to different radiation doses, oncogematology patients) had been summarized and analyzed. The conclusion concerning possibilities and limitations of FISH technique usage for retrospective biodosimetry of human radiation exposure has been presented.

Aim: Although acute myelogenous leukemia (AML) arising after radiation exposure is considered to be secondary, little is known about the molecular mechanisms by which the radiation induces the leukemogenic phenotype. The aim of the study was to analyze whether the MLL translocations are as frequent in radiation-associated AML as in spontaneous AML cases., Methods: Sixty one AML samples obtained at diagnosis were analyzed for the presence of MLL abnormalities using fluorescent in situ hybridization and/or reverse transcription polymerase chain reaction. Of these patients, 27 had experienced radiation exposure due to the Chernobyl accident, 32 were non-irradiated (spontaneous AML), and 2 developed therapy-related AML after chemotherapy with topoisomerase II inhibitors., Results: MLL gene translocations were detected in both groups of spontaneous and therapy-related AML (1/32 and 1/2 cases respectively). The sole MLL rearrangement found in the group of radiation-associated AML patients was a duplication of the gene., Conclusion: Our data preclude the involvement of MLL gene translocations in radiation-induced leukemogenesis, but support the assumption that loss and gain of chromosomal material could be crucial in the leukemogenesis of AML patients with the history of radiation exposure due to the Chernobyl accident.

The morphofunctional state of hemopoiesis in patients who suffered from the Chernobyl accident was investigated. The activation of proliferative activity of erythroid progenitor cells simultaneously with a decrease in the resistance of erythrocytes and an increase in the levels of methemoglobin, HbF, and an increase in the number of HbF-containing cells in peripheral blood were detected. Main trends in the development of adaptative processes in the erythron system as a response to affect of low doses of radiation were determined on cellular and molecular levels.

Prolonged cultivation of K-562 human leukemic cells in plastagar diffusion chambers inserted into the peritoneal cavities of CBA mice indicated erythroid commitment and differentiation of cells. It was shown by the method of dot-hybridization with the use of oligonucleotide probe the presence of mRNA of globin in these cells.

A total of 18 patients with acute leukemia from those exposed to radiation when they had liquidated the consequences of the accident at the Chernobyl Atomic Power Station were examined. Sixteen of them had acute myeloblast cell leukemia, one had acute lymphatic leukemia, and another one had acute promyelocytic leukemia. A complex of hematological, immunocytological, and cytochemical findings, as well as leukocytic and erythrocytic genetically determined antigen typing were used for diagnosis. Signs of severe vegetovascular dystonia was found to be followed by acute leukemia in all the patients. A relationship was established between the therapy resistance and the HLA antigen homozygosity in combination with genetically-determined low reactivity. There were no significant differences between the parameters in question and those in patients with acute leukemia who had been studied in the preaccident period.

The qualitative and quantitative parameters of hemopoiesis and immunocompetent cells were studied in 365 children in the early "iodine" and long-term periods after the accident. In accordance with primary response, a group consisting of 28 subjects with so-called "radiation injury" was distinguished. The early period was marked by unusual qualitative and quantitative hematological changes which, however, did not lead to the development of the hematological syndrome characteristic of acute radiation injury. Certain deviations that corresponded to somatic pathology were detectable at the late periods after the accident.

Immunoassays of the 1st and 2nd levels were investigated in patients with acute radiation disease 12-36 mos. after irradiation. A decrease in the level of helper/inducer T-lymphocytes, a tendency to a decrease in the number of suppressors and an increase in the number of theophylline-stimulated T-cells in patients with acute radiation disease of the 3rd degree were observed. Control of the immune status of this group of patients is recommended.

Collagen I was isolated from human bone tissue and from mice bone tissue of the AKR-50 strain at the pronounced stage of leukosis. Dissimilarity of native and leukemic collagens was exhibited after evaluation of their amino acid composition, electrophoretic mobility in polyacrylamide gel containing SDS, content of the carbohydrate moiety as well as of isopoints, elution profiles in reverse-phase chromatography and gel filtration and electron microscopy of SLS-crystallites. Impairments of collagen processing in leukemia appear to be responsible for its alterations in structure and properties.

Colony- and cluster-forming ability of bone marrow cells in agar medium was studied in 6 healthy children and 13 children suffering from acute leukosis. During acute stages of the diseases colonies and clusters in agar medium consisted of granulocytic cells and their quantity depended on the safety of predecessors of bone marrow granulopoiesis. In contrast to acute lymphoblastic leucosis, acute myeloblastic leucosis is characterized by a sharp growth of cluster-forming and a less intensive growth of colony-forming ability of hemopoietic cells. The bone marrow remission is marked by the growth of diffusive and mixed colonies and clusters, consisting of granulomonocytic elements. The low level of colony- and cluster-formation in the medium at the last stage of therapy (induction of acute leukosis remission) correlated well with positive indices of treatment effectiveness. The results obtained during the cultivation of blood-forming cells in the agar medium may be also used for diagnosis and prognosis.