[Features of allele polymorphism of genes involved in homocysteine and folate metabolism in patients with atherosclerosis of the lower extremity arteries].

Under study were features of allele polymorphism of genes of methylenetetrahydrofolate reductase (MTHFR C677T and A1298C), methionine synthase (MS A 2756G), methionine synthase reductase (MTRR A66G) and methylenetetrahydrofolate dehydrogenase (MTHFD G1958A) in patients with atherosclerosis of the lower extremity arteries (ALEA). Patients with hyperhomocysteinemia (HHcy) had statistically significant increase of allele MTHFR 677T and MTRR 66GG as compared both with the control group and with the group of patients without HHcy. It suggests that polymorphism of genes involved in homocystein and folate metabolism might affect the risk of HHcy in patients with ALEA.

[Study of lactate dehydrogenase isoenzyme in the cerebrospinal fluid of patients with acute lymphoblastic leukemia].

Lactate dehydrogenase isoenzymes were profiled in cerebrospinal fluid in patients with acute lymphoblastic leukemia. Cytological examination of patients with central nervous involvement identified isoenzyme 5 which failed to show after selective treatment. The isoenzyme's detection in cerebrospinal fluid without typical cytological characteristics may be regarded as a precursor of such complication. Hence, this may be used as an additional criterion for refinement of cytological evidence employed in diagnosis and prognosis of central nervous involvement in patients with acute lymphoblastic leukemia.

[Genetic determinants of hereditary thrombophilia in pathogenesis of venous thrombosis]. / Geneticheskie determinanty nasledstvennoi trombofilii v patogeneze venoznogo tromboza.

AIM: To study the role of genetic determinants of hereditary thrombophilia in pathogenesis of various clinical manifestations of venous thrombosis in the citizens of the North-West Region of Russia. MATERIAL AND METHODS: Mutations of the genes of factor V (FV Leiden), prothrombin (G20210-A) and polymorphism C677-T in the gene of methylentetrahydrofolate reductase (MTHFR) were detected using polymerase chain reaction (PCR) with a following restriction analysis of PCR product in 183 patients with venous thrombosis (115 with isolated thrombosis of the deep veins and 68 with thromboembolism of the pulmonary artery). RESULTS: It was established that mutation FV Leiden is a significant risk factor of deep vein thrombosis in the legs and postthrombotic disease, but this mutation is weakly associated with pulmonary artery thromboembolism (PAT). An essential PAT risk factor is carriage of the variant prothrombin G20210A. CONCLUSION: Determination of prothrombotic genotypes is a key factor of treatment efficacy and prevention of life-threatening thromboembolic complications.

[Anticytosine therapy for primary myelodysplastic syndrome]. / Antitsitokinovaia terapiia bol'nykh pervichnym mielodisplasticheskim sindromom.

A pilot study of anticytocine therapy (PCD) was carried out in 9 patients with primary myelodysplastic syndrome (MDS). After 12 weeks, 2 patients (22%) showed hematological response cutting down the need for washed-out erythrocyte transfusions by half or more. MDS progression at different stages was reported in 4; transformation to acute myeloleukemia--2. PCD treatment is indicated in MDS patients, aged over 65, with a less than 10% blast level of the bone marrow, and without any risks of tumor progression.

HLA-DR4-Ala74 beta is associated with risk and poor outcome of severe aplastic anemia.

Severe aplastic anemia (SAA) is a heterogeneous hematological disorder with a high mortality. Genetic predisposition has been shown to play a role in a considerable proportion of SAA cases. For instance, the human lymphocyte antigen HLA-DR2 has been repeatedly demonstrated to be over-represented in SAA patients. In this paper, we expand on the evidence for the contribution of HLA polymorphism in the susceptibility to SAA, which was obtained using the "high-resolution" technique of HLA-DRB1 subtyping. The DRB1*1501 allele appeared to be responsible for the predominance of DR2 specificity in SAA patients and was the most significant risk factor for this disease. It was observed in 23/44 (52.3%) patients versus 22/100 (22.0%) donors [odds ratio (OR) = 3.9; 95% confidence interval (CI): 1.8-8.3; P = 0.0005, corrected P (Pc) < 0.05]. In addition, DRB1*04 alleles also displayed non-random distribution in the SAA group. In particular, DRB1*04 variants coding for alanine at position 74 of the DR beta 1 chain (HLA-DR4-Ala74 beta subtype) were detected in all 13 DR4-positive SAA patients but only in 15/24 (62.5%) controls (OR = 16.6; 95% CI: 0.9-312.0; P = 0.015). Multiple comparison analysis confirmed that the HLA-DR4-Ala74 beta subtype confers susceptibility to SAA independently from the DRB1*1501 allele. Finally, examination of the clinical records has shown that the HLA-DR4-Ala74 beta subtype is associated with poor outcome of SAA.

[Diagnostic and prognostic significance of some laboratory indices of cerebrospinal fluid in multiple myeloma]. / Diagnosticheskoe i prognosticheskoe znachenie nekotorykh laboratornykh pokazatelei spinnomozgovoi zhidkosti u bol'nykh mnozhestvennoi mielomoi.

beta 2-microglobulin (beta 2-m) and certain other characteristics of liquor have been followed in the course of multiple myeloma (MM) versus severity of the disease and its response to chemotherapy, to establish the diagnostic and prognostic value of these indices. Lymphocytes and plasma cells were found in most of 17 patients under study, who were resistant to chemotherapy and relapsed; they revealed regular rise in liquor pressure and cytosis. Similarly, raised concentrations of beta 2-m were registered. However, there was no correlation between blood serum-beta 2-m levels and that in cerebrospinal fluid (CSF) nor between CSF-beta 2-m concentration, on the one hand, and such parameters as CSF-total protein, blood serum-beta 2-m and pathologic cytosis, on the other. In advanced MM, CSF-beta 2-m levels were, as a rule, abnormally higher. Our data suggest that, in addition to assaying CSF-total protein, severity of cytosis and liquor pressure as well as morphological identification of detected cells, beta 2-m levels be followed in the course of the disease in MM patients, to make therapeutic correction possible. Moreover, CSF-beta 2-m level dynamics can be used as another factor of prognosis.

[Diagnostic and prognostic significance of blood-serum ceruloplasmin, acetylcholinesterase and total proteolytic activity in patients with multiple myeloma]. / Diagnosticheskoe i prognosticheskoe znachenie opredeleniia tseruloplazmina, atsetilkholinesterazy i obshchei proteoliticheskoi aktivnosti v syvorotke krovi bol'nykh mnozhestvennoi mielomoi.

The study included 82 patients with multiple myeloma (MM) to evaluate the diagnostic and prognostic significance of correlation between of levels of ceruloplasmin (CP), acetylcholinesterase (ACE) and total proteolytic activity (TPA) in blood serum and immunochemical pattern, tumor mass and response to chemotherapy. It was shown that CP, ACE and TPA determination may be used as additional markers to confirm therapeutic benefits, to timely detect relapse and to verify resistance to chemotherapy. It was also demonstrated that resultant decrease in CP and TPA and increase in ACE levels are reliable indicators of changes developing in MM course and remission fulfillment. High values for CP and low ones for ACE point to pathological activity. Both diagnostic and prognostic significance of the indices has been shown. The highest levels of CP and TPA in blood serum were identified in cases of A-myeloma and myeloma of Bence Jones while ACE concentrations in those patients were lower than in G-myeloma which correlated with median survival. Application of said assays might improve diagnosis, management and prognosis of MM.

[Detection of chromosome translocation t(9;22) using RT-PCR]. / Vyiavlenie khromosomnoi translokatsii t(9;22) metodom RT-PTSR.

The investigation deals with a simplified modification or molecular-genetic detection of translocation t(9;22) using a combination of reverse transcription and polymerase chain reactions (RT-PCR). Unlike the available protocols, analysis is carried out using one enzyme--TET-Z polymerase--(instead of two) which has both revertase and DNA-polymerase activities. The present modification is highly sensitive, less time-consuming and cheaper. The method has proved useful for both diagnosing t(9;22) translocation and diagnosing and monitoring minimal residual disease remaining after marrow transplantation.

[Prognostic value of some biochemical and immunologic factors in the management of multiple myeloma]. / Prognosticheskaia tsennost' nekotorykh biokhimicheskikh i immunologicheskikh faktorov pri mnozhestvennoi mielome.

The results of examination of 58 patients in the course of multiple myeloma (MM) are discussed. Prognosis for LDH, beta2-m and FNO-alpha administration was evaluated versus immunochemical pattern, tumor mass and response to different chemotherapeutic regimens. High prognostic value of serum levels of LDH, beta2 and FNO-alpha in MM patients was observed. The method may be useful in carrying out chemotherapy and detecting recurrences and chemoresistance.

[The importance of studies of the glycosaminoglycans in the peripheral blood leukocytes of patients in the diagnosis of the phases of chronic myeloleukemia]. / Znachenie issledovanii glikozaminoglikanov leikotsitov perifericheskoi krovi bol'nykh v diagnostike faz khronicheskogo mieloleikoza.

To elucidate new diagnostic markers of chronic myeloid leukemia (CML) phases, we investigated quantitative and qualitative composition of glycosaminoglycans (GAG) of leukocytes from peripheral blood of 72 patients. Chronic CML phase was characterized by elevated GAG levels (2 times compared to normal values), weakening of anionic properties of chondroitin sulfate (CS) and high amount of heparan sulfate (HS). In CML transformation in the progressive phase overall concentration of GAG grew still higher, GAG fraction composition changed. In the blast crisis there was a sharp fall in the overall GAG, new electrophoretic fractions emerged. In the myeloid variant of the crisis an additional GAG component appeared (GAG-m), whereas in the lymphoid variant another component was found (GAG-1). It is suggested that the number and composition of GAG in peripheral blood leukocytes may serve markers of CML phase.

A rapid RT-PCR based method for the detection of BCR-ABL translocation.

AIMS: To optimise a one step reverse transcriptase polymerase chain reaction (RT-PCR) protocol for BCR-ABL chimaera detection. METHODS: Compared with published RT-PCR procedures, this novel approach has at least two advantages. First, the same enzyme is used for both reverse transcription and PCR. Second, amplification of the target (BCR-ABL chimaera) and control gene (ABL) is performed simultaneously in the same tube. RESULTS: On testing 40 chronic myelogenous leukaemia patients and 10 healthy donors there was a specificity for the newly developed technique. In addition, dilution experiments demonstrated that the protocol was highly sensitive. CONCLUSIONS: The suggested one step PCR strategy is a simple and reliable way to reveal BCR-ABL chimaeras.

[Is the cytogenetic verification of chronic myeloleukemia always obligatory?]. / Vsegda li obiazatel'na tsitogeneticheskaia verifikatsiia khronicheskogo mieloleikoza?

To elucidate feasibility of accurate diagnosis of chronic myeloid leukemia (CML) without cytogenetic and molecular-genetic investigations as well as to specify CML diagnostic criteria, clinicohematological parameters were compared in two groups of patients: with Ph'-chromosome and/or rearrangement of fragment bcr (group 1), with unknown karyotype in whom detection of bcr fragment rearrangement was not made. Clinicohematological parameters in both groups were close in absolute value and underwent parallel changes in the course of leukemia progression. In group 1, patients in progressive and blastic phase compared to patients in chronic phase had a 14-fold increase in the number of additional cytogenetic anomalies. In patients with tumor transformation fragment bcr underwent rearrangement according to type B2A2. Thus, the diagnosis of typical CML variants is feasible without detection of Ph'-chromosome and/or rearrangement of bcr fragment. It is especially true and essential for patients in the chronic phase. The data obtained provide more accurate diagnostic criteria of CML.

[The prognostic significance of the beta 2-microglobulin level of the blood serum in multiple myeloma patients]. / Prognosticheskoe znachenie urovnia beta 2-mikroglobulina v syvorotke krovi bol'nykh mnozhestvennoi mielomoi.

Pretreatment and treatment content of beta 2-microglobulin in blood serum was measured in 52 patients with multiple myeloma stage IIA, IIB, IIIA and IIIB. The above concentrations before treatment correlated with the disease stage, rose in the exacerbation, remained unchanged in stabilization, fell in effective therapy and returned to normal values in remission. However, in pronounced renal insufficiency (stage IIIB) the remission beta 2-microglobulin levels remained high. A 6 mg/l concentration proved critical for survival. The survival median at pretreatment beta 2-microglobulin content above 6 mg/l was 14 months, under 6 mg/l--49 months. Routine estimation of beta 2-microglobulin levels is necessary for better control over multiple myeloma treatment, timely change of cytostatic drugs.

[Experience with the use of reaferon (alfa 2-interferon) for treating patients with chronic myeloleukemia]. / Opyt primeneniia reaferona (al'fa 2-interferona) dlia lecheniia bol'nykh khronicheskim mieloleikozom.

The effectiveness of domestic alpha 2-interferon preparation reaferon was studied in vivo and in vitro for eradication of pathological hemopoietic clone in chronic myeloid leukemia. Reaferon administration for 1-30 months produced cytogenetic remission in 7%, hematological remission in 21%, partial hematological remission in 36% of the patients. Reaferon is indicated in chronic myeloid leukemia without splenomegaly. In the disease progression reaferon is uneffective. Mechanism of reaferon therapeutic action comprises three components: a direct antiproliferative effect on hemopoietic precursor cells, activation of cellular immunity, an effect on stem cell microenvironment.

[Thymidine kinase activity in leukocytes from patients with chronic myeloid leukemia at various periods in the disease]. / Aktivnost' timidinkinazy v leikotsitakh bol'nykh khronicheskim mieloleikozom v razlichnye periody zabolevaniia.

The level of thymidine kinase activity in the premature leukocytes of patients with chronic myeloleukemia during the stable phase was shown to serve as a measure of the disease development. Considerable variations in thymidine kinase activity in blast cells in myeloid and lymphoid blast crises demonstrated that analysis of the enzyme activity might be used in the biochemical diagnosis of blast crisis in chronic myeloleukemia simultaneously with the enzymes of purine metabolism--ADA and PNP. During cell differentiation, the activity of thymidine kinase was decreased and in the myeloid cells the enzymatic activity was much higher of that in lymphoid cells as shown by investigations using blast cells of patients with blast crisis in chronic myeloleukemia, cells K-562, thymocytes, spleen and peripheric blood lymphocytes. Isozyme thymidine kinase I was mainly responsible for the rate of enzymatic activity in premature leukocytes of patients with chronic myeloleukemia regardless of the disease stage.

[Polyadenylate polymerase in blood serum from hemoblastosis patients]. / Poliadenilatpolimeraza v syvorotke krovi bol'nykh gemoblastozami.

Activity of polyadenylate polymerase was studied in blood serum of patients with some forms of leukemia. In acute form of leukemia, the activity of the polymerase was increased 3-fold in blood serum as compared with normal state. Under conditions of chronic myeloleukemia the enzymatic activity was altered depending on the disease step with increase in leukemia development and reached the maximal values in blast crisis. The maximal enzymatic activity was found in myelodysplastic syndrome in patients with refractory anemia and excess of blasts.

[Distribution of alleles of DRB, DQB and DQA loci of major histocompatibility complex in healthy donors of Saint Peterburg]. / Osobennosti raspredeleniia allelei lokusov DRB, DQB i DQA glavnogo kompleksa gistosovmestimosti u zdorovykh donorov Sankt-Peterburga.

The distribution of alleles of DRB, DQB and DQA loci was investigated in 118 healthy donors from St. Petersburg. Said alleles were identified by the restriction fragment length polymorphism method using DRB, DQB and DQA probes after TaqI digestion and Southern blotting. The frequencies of said alleles were compared with those of a cohort of healthy donors living in Germany. A significantly higher frequency of DRB-17-2 was identified in the Russian donors, as compared with the German counterparts. Also, the Russian donors revealed a decrease in the frequency of DQB-1 (P = 0.01) and an increase in that of DQB-X allele (P = 0.05). An analysis of DRB, DQB and DQA locus a alleles adhesion in a sample of donors from St. Petersburg showed it to agree, in a large proportion of cases, with the literature data. A parallel study of immunological specificity of DR antigen was undertaken to compare the results of genetic and serologic typing; errors in DR specificity identification were found to have been made in 17.0%.