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Background. 6-mercaptopurine (6-MP) is a drug that is included in the treatment protocols for children and adults with acute lymphoblastic leukemias/lymphomas (ALL/LBL). It is known that individual differences in 6-MP tolerance can be explained by the TPMT and NUDT15 polymorphisms.Aim. To determine 6-MP toxicity profile in adult patients with Ph-negative ALL/LBL treated by ALL-2016 protocol, depending on the TPMT and NUDT15 polymorphisms.Materials and methods. The study included 54 adult patients with Ph-negative ALL/LBL (40 male and 14 female). The median age was 31 (18-51) years. T-ALL/LBL was diagnosed in 29 patients, B-ALL/LBL - in 22, acute leukemia with a mixed immunophenotype - in 3. All patients received treatment according to the multicenter study ALL-2016 (ClinicalTrials.gov, NCT03462095). polymorphisms in NUDT15 (*2, *3) and TPMT (*2, *3A, *3B, *3C) genes were detected using the allele-specific real-time polymerase chain reaction. Genomic DNA was extracted from patients peripheral blood samples. On the induction and consolidation therapy by the protocol, the received and proper 6-MP doses were calculated for all the patients. Drug toxicity was evaluated based on clinical and laboratory data.Results. TPMT and NUDT15 polymorphisms were detected in 11 (20 %) patients, more often in B-ALL - 7 (32 %) of 22 (p TPMT, NUDT15 polymorphisms only at consolidation IV (p = 0.01). we didn't find a correlation between the 6-MP toxicity and the polymorphisms in our patients (p >0.05).Conclusion. There were no differences in the received dose of 6-MP and the incidence of toxicity in adult patients between Ph-negative ALL/LBL with or without TPMT and NUDT15 polymorphisms treated according to ALL-2016 protocol (p >0.05). further studies including evaluation of 6-MP metabolites concentrations are required for a more complete understanding of the metabolism of this drug.

Background. In addition to anti-HLA-I and anti-HPA-antibodies and specific cytotoxic T-lymphocytes, another cause of immune refractoriness to donor's platelet transfusions could be a platelet-associated different classes immunoglobulins PAIg (G, M, A) and C3 / C4‑components of complement system (PAC3, PAC4). These markers can be detected by flow cytofluorometry of double-stained platelets. The fixation density of immunoglobulins and components of complement systems were measured by the mean fluorescence intensity (MFI).Objective: to study additional factors that aggravate the course of refractoriness to donor's platelet transfusions in patients with aplastic anemia (AA) and hemoblastosis.Materials and methods. 77 patients (AA – 47, myelodysplastic syndrome (MDS) – 10, acute myeloid leukemia (AML) – 20) admitted to National Research Centre for Hematology during 11.09.2016–04.28.2018 were enrolled in the study. M / f ratio was 33 / 44, median age was 36 yrs. (19–71 yrs.). Plasmapheresis and cross-matching for PRP selection were used for patients with refractoriness to donor's platelet transfusion. PAIg (G, M, A) and PAC3 / C4 detection and density (MFI) were evaluated in all patients by flow cytofluorometry of doublestained platelets (CD41a-PE; IgA, M, G-FITC; C3 / C4‑FITC) and MFI measurement. Patients with AA were investigated on different stages of therapy and if refractoriness to donor's platelet transfusion is developed. Blood donors (n = 28) MFI measurement results were established as negative control.Results. It was found that MFI PAIgG/M/А and PAC3/С4 was higher in all groups of the patients (АА, MDS, AML), as compared with donors. MFI of PAIgM and PAIgA in patients were significant higher than MFI of PAIgG and PAC3 / C4. Combination of PAIgM / A, PAIgM / C3 / C4 and PAIgA / C3 / C4 were more frequent. Multiple transfusions of PRP were associated with PAIgA and PAC3 detection. Development of refractoriness to donor's platelet transfusions was accompanied by alloantibodies (HLA-I, HPA) and PAIgM, PAC4 detection. In patients of AA group during development of refractoriness to donor's platelet transfusions and multiple infection complications the high density of PAIgM and PAIgA were identified. Relapse of AA was accompanied MFI of PAC3 density increment.Conclusion. In addition to application of a certain transfusion therapy algorithm it is also necessary to detect PAIg (G, M, A) and PAC3 / C4 for prediction of severe refractoriness to donor's platelet transfusions.

Background . Myelodysplastic syndromes (MDS) are a heterogeneous group of clonal diseases of the hematopoiesis system characterized by dismyelopoiesis and cytopenia, the presence of cytogenetic aberrations and a high risk of transformation into acute myeloid leukemias. Diagnosis of MDS requires a comprehensive approach and mandatory performance of cytological, cytochemical and cytogenetic studies of bone marrow aspirate, as well as histological examination of trephine biopsy. However, in some cases it is necessary to undergo a diagnostic test that would allow verification of the MDS. The study of bone marrow aspirate by multicolor flow cytometry (MFC) can be considered as an additional diagnostic criterion in the diagnosis of MDS.The objective of the study was to estimate the incidence of myelodysplastic features in patients with various forms of MDS by the MFC method. Materials and methods . The study included 79 patients with MDS: 8 with MDS with 5q deletion, 33 with MDS without excess blast cells and 38 with excess of blasts. A bone marrow aspirate test was performed by 6-color flow cytometry. The control group included 35 donors of allogeneic bone marrow. The analysis resulted in a conclusion on the Ogata score scale, the Wells prognostic scale and the combined Ogata–Wells scale. When using the screening method, the presence of two or more cytometric signs of MDS was detected in 60 (75.9 %) of 79 MDS patients. Wells score was higher in MDS group with an excess of blast than in others. Using the combined Ogata–Wells scale, cytometric aberrations were found in 70 (88.6 %) of 79 MDS patients. In patients with MDS with an excess of blasts, the incidence of increased CD34+ and/or CD117+ myeloid cells was higher than in MDS patients without an excess of blasts and an MDS with a 5q deletion. The frequency of abnormal cytometric parameters (anomalous expression of CD34, CD117, CD56+ myeloblasts) in these groups did not differ. In patients with isolated 5q deletion and MDS without excess of blasts, an increased proportion of CD7+CD34+ cells was more often detected than in MDS with an excess of blasts.Conclusion . Thus, cytometric abnormalities in MDS are common, even in patients without excess of blasts. The MFC method can be used as an additional diagnostic method in the initial diagnosis of MDS.

Refractoriness to transfusions of platelet concentrates (PC) adversely affects the conduct of complex therapy in hematological patients. Individual selection of platelets is recommended for such patients. In cases of high degree of alloimmunization with the formation of polyspecific antibodies, when individual selection is difficult, procedures plasmapheresis (PPs) is included in the treatment program.Aims: to evaluate the effectiveness of PC transfusions by individual selection in patients refractory to transfusions and the use of PPs as a second line therapy in combination with individual platelet selection.Materials and methods: from September 2015 to December 2017, 91 patients with refractory to PC transfusions from 1263 patients who received PC transfusion were observed in the center’s clinics. The median age was 43 (18–71) years. M/F – 38/53. Patients: 20 – aplastic anemia (AA), 17 – myelodysplastic syndrome (MDS), 45 – acute myeloid leukemia (AML), 9 – acute lymphoblastic leukemia (ALL). All patients underwent PC transfusion by individual selection (HLA/HPA) Immucor’s Capture-P solid phase technology. In 28 (30 %) of 91 patients, due to the inability to select, there was a need for PP as a second line therapy. Patients: AA – 4 (20 %); MDS – 8 (47 %); AML – 12 (26 %); ALL– 4 (44 %). The median age was 48 (23–71) years. M/F – 8/20. From 2 to 15 procedures were performed (on average – 6) for each patient. All patients received PC transfusions by individual selection by cross-matching immediately after the PP procedure. The efficacy of PC transfusions was assessed by Absolute Platelet Increment (API) and Corrected Count Increment (CCI), relief of hemorrhagic syndrome.Results: in 26 of 28 refractory to PC transfusions patients, in the absence of compatible donor platelets, carrying out PPs in combination with subsequent individual platelet selection promoted relief of hemorrhagic syndrome, increase in API from 3.3 × 109/L at 29.5 × 109/L and CCI from 1.3 to 10.7. Against the background of PPs, combined with individual selection, the degree of alloimmunization (the percentage of incompatible pairs) decreased on average: AA (n = 4) – from 91.7 to 50.2 %; MDS (n = 8) – from 89.6 to 31.6 %; AML (n = 12) – 86.0 to 40.5 % and ALL (n = 4) – from 91.7 to 37.7 %. In 2 patients with a high degree of alloimmunization and after carrying out PPs, it was not possible to select compatible platelets, PC transfusions were ineffective (API = 5 × 109/L, CCI = 1), and hemorrhagic syndrome was not completely managed, but its severity was reduced.Conclusions. With the development of refractoriness to PC transfusions and the ineffectiveness of individual platelet selection, PPs should be used as the second line of therapy, which, combined with individual selection, increases the likelihood of compatible donor-recipient pairs and increases the clinical efficacy of PC transfusions. When PPs is ineffective in combination with individual selection, it is necessary to exclude the syndrome of increased consumption and other mechanisms of refractoriness.

Objectives. The objective of this study was to evaluate epidemiology of febrile events (FE) and efficacy of antibiotic treatment in neutropenic patients with newly diagnosed acute myeloid leukemia (AML) with and without colonization of gut by extended-spectrum β-lactamaseproducing Enterobacteriacae (ESBL-E). Materials and methods. The prospective study (2013–2015) included 66 patients with AML. These patients received 208 chemotherapy cycles within 6 month. Rectal swabs were obtained from all patients prior to antibiotic administration. ESBL-E were isolated on chromogenic ESBL selective medium CHROMagarТМESBL (CHROMagar, France) and confirmed by double disk synergy test. Results. FE occurred in 193 (93 %) of chemotherapy cycles. The analysis was performed in 173 FE, including 68 – with colonization and 105 – without colonization with ESBL-E. Epidemiology of FE was similar in patients colonized by ESBL-E and non-carriers group, except cases of bacteremia, caused by ESBL-E that occurred only in patients colonized by the same bacteria (7.5 %; p = 0.009). Patients colonized by ESBL-E and non-carriers had comparable efficacy of first-line non-carbapenem regimens (38 % vs 44 %), rate of carbapenem administration (62 % vs 55 %), efficacy of carbapenems alone (36 % vs 52 %) and in combination (64 % vs 41 %), duration of all antibiotics (14 days vs 13 days) and carbapenems (10 days vs 10 days). All cases of bacteremia caused by ESBL-E were successfully treated by carbapenems. Conclusion. Colonization of gut with ESBL-E is a predictor of bacteremia caused by the same bacteria. There were no differences in the use of antibiotics in patients colonized by ESBL-E and non-carriers group.