[Molecular and biological properties of the African swine fever virus (Asfarviridae: Asfivirus) isolate ASF/Tatarstan 20/WB-12276].

INTRODUCTION: Up-to-date data and full characterization of circulating ASFV isolates play a crucial role in virus eradication and control in endemic regions and countries. The aim of the study was to evaluate and characterize the molecular and biological properties of the ASFV isolate ASF/Tatarstan 20/WB-12276, conduct phylogenetic analysis, and compare the results with isolates circulating in Europe and Asia. MATERIALS AND METHODS: For bioassay, eight heads of the Large White pigs weighing 15-20 kg/head were used. Detection of specific anti-ASFV antibodies by ELISA and immunoperoxidase method. Detection of ASFV genome was performed by qPCR. Isolation of ASF/Tatarstan 20/WB-12276 and determination of titer were performed in pig spleen cell culture. Sequencing was carried out by the Sanger method. RESULTS: The virus was characterized as highly virulent and capable of causing acute to subacute forms of ASF. Phylogenetic analysis revealed substitutions in the genome of the ASF/Tatarstan 20/WB-12276 isolate (IGR/I73R-I329L and I267L markers) that supported the clustering of the studied variant with isolates prevalent in most of Europe and Asia. CONCLUSION: For the first time, the molecular and biological properties of the ASF/Tatarstan 20/WB-12276 virus isolate taken from a wild boar shot on the territory of the Republic of Tatarstan were studied and analyzed.

[The prognostic role of determining spontaneous leukocyte DNA damage and that occurring during chemotherapy in leukemia patients]. / Prognosticheskaia rol' opredeleniia spontannykh i voznikaiushchikh v protsesse khimioterapii povrezhdenii DNK leikotsitov bol'nykh leikozami.

The authors studied spontaneous DNA damages and the extra plan synthesis of DNA in various leukemias (chronic lympholeukemia, chronic myeloleukemia, acute leukemia) to predict the natural course of leukemias and the efficiency of chemotherapy. The level of genuine DNA breaks, which had been determined by the nic-translation test, was lower in peripheral blood cell DNA in all leukemias than that in the lymphocytes and granulocytes from donors. On the contrary, the levels of alkaline-labile DNA sites and nuclear matrix protein-bound DNA in the mature and maturing leukemia cells are high and decrease with progression of chronic lympholeukemia and chronic myeloleukemia. The extra plan DNA synthesis largely reflects the cellular levels of genuine DNA breaks, and the ultraviolet-induced DNA repair capacity is the highest in the lymphocytes in chronic lympholeukemia and particularly in the blasts in acute leukemia. The efficiency of chemotherapy was predicted during chemotherapy from the intensity of formation of genuine blast cell DNA breaks.

[Ultrastructural organization and argentophilic areas of the lymphoblastic nucleoli in patients with acute lymphoblastic leukemia]. / Ul'trastrukturnaia organizatsiia i argentofil'nye zony iadrushek limfoblastov bol'nykh ostrym limfoblastnym leikozom.

The ultrastructural organization and the state of silver-staining nucleoli (Ag-NOR) of peripheral blood and bone marrow lymphoblasts of 10 patients with acute lymphoblastic leukemia (ALL) were studied in the complex with the biochemical assay of the ribosomal levels in the cellular cytoplasm of these patients. In the majority of the patients investigated the morphological picture of the nucleolus and the character of Ag-NOR have evidenced the high functional activity of pRNA-synthesizing apparatus and correlated with the high ribosomal level in the cytoplasm of the cells studied. Significant accumulation of granular components (RNP-particles) in the nucleoli of the lymphoblasts, reflecting disorders in their transport from nucleus to cytoplasm, were detected only in 2 patients.

[Ultrastructural organization of nucleoli and ribosome level in lymphocytes at different stages of maturation in patients with chronic B-cell lymphocytic leukemia]. / Ul'trastrukturnaia organizatsiia iadryshek i uroven' ribosom v razlichnykh po stepeni zrelosti limfotsitakh krovi bol'nykh B-kletochnym khronicheskim limfoleikozom.

To clarify the role of the ribosome apparatus in the differentiation and maturation, the nucleolar ultrastructure, ribosome level and maturation stages of lymphocytes and their phorbol ester-induced differentiation capacity were studied immunologically, cytochemically and electron-microscopically in 16 patients with B-chronic lymphoid leukemia (B-CLL) and in one with small lymphocyte lymphoma. Most B-CLL cases had the clones mainly consisting of cells at an intermediate maturation stage. The cases with immature and mature lymphocyte clones were observed more rarely. The acid phosphatase activity did not correlate with the maturation stages of the lymphoid cells. The nucleolus activity ranged from virtually zero to high one. In general, there was a relationship between the ribosome content and nucleolus activity, although there were exceptions suggesting the impaired ribosome biogenesis. There was no correlation between the ribosome content and maturation degree of B-CLL lymphocytes and their phorbol-ester-induced differentiation capacity.

[Ribosome content, spontaneous damage and repairing DNA synthesis in patients with chronic B-lymphocytic leukemia depending on stage and form of disease]. / Soderzhanie ribosom, spontannye povrezhdeniia i reparatsionnyi sintez DNK u bolnykh B-khronicheskim limfoleikozom v zavisimosti ot stadii i formy zabolevaniia.

The ribosome content (84 patients), DNA alkaline-labile sites or strand breaks (21 patients) and unscheduled DNA synthesis in lymphocytes under normal and B-cell chronic lymphocytic leukemia (B-CLL) were studied. It has been shown that the alkaline-labile DNA damages increased about 3-5 fold while the ribosome content was normal at an early (corresponding to Rai's O-I stages) and an advanced (II-III stages by Rai) stages of B-CLL. In the course of B-CLL, the DNA strand breaks and number of ribosomes per cell decreased with the correlation coefficient r = 0.72. The slowing down of the unscheduled DNA synthesis at the advanced and terminal (IV stage by Rai) B-CLL stages does not correlate with the DNA strand breaks. In the advanced stage with its established clinical variability of forms, when an adequate treatment was necessary, the lymphocyte ribosome levels drop following the expansion of main compartments involved by the leukemic spread.

[Ribosome levels in T- and B lymphocytes in B-cell chronic lymphocytic leukemia]. / Soderzhanie ribosom v T- i B-limfotsitakh pri B-khronicheskom limfoleikoze.

The levels of ribosomes in B-chronic lymphocytic leukemia (B-CLL) T lymphocytes have been shown to be within normal whereas those in B-CLL B-lymphocytes to vary from normal to fairly low levels. There was no dependence of the ribosome content on the maturation of B lymphocytes as it was established by levels rosetting with mouse red blood cells.

[Characteristics of the silver-stained nucleoli in blast elements of various diameters from patients with acute leukemia]. / Kharakteristika okrashennykh serebrom iadryshek v blastnykh élementakh raznogo diametra bol'nykh ostrymi leikozami.

Using silver staining, a study was made of the activity of ribosomal cistrons (RC) in proliferating and non-proliferating subpopulations of bone marrow and peripheral blood blasts, taken from 33 patients with acute non-lymphoblastic leukemia and from 17 patients with acute lymphoblastic leukemia. All the blasts including non-proliferating ones were revealed to synthesize ribosomal RNA (rRNA). The RC activity of large blasts was higher than that of small ones. A significant difference (P less than 0.01) in RC activity between lymphoid and non-lymphoid blasts was found. The relation between nucleolar organizer activity of blasts and their proliferating potentials is discussed, in addition to a possibility for practical use of the above mentioned data.

[Characteristics of ribosomal RNA synthesis in the blood lymphocytes of chronic lympholeukemia patients]. / Osobennosti sinteza ribosomnoi RNK v limfotsitakh krovi bol'nykh khronicheskim limfoleikozom.

A study was made of the number of silver grains over the nucleoli and of the content of ribosomes in the lymphocyte cytoplasm for six healthy persons and for 20 patients with chronic lymphatic leukemia. Besides ratios of compact, nucleolonemic and ring types of nucleoli were evaluated in addition to counts of the specific radioactivity of mature 28S rRNA in lymphocytes. In the majority of cases examined, cells with 1 or 2 nucleoli of compact and nucleolonemic types were seen dominating. The number of silver grains over the nucleoli in the control healthy persons did not differ from that in patients who did not receive any treatment, which contrasted with high value grain counts in the treated patients. The lymphocyte ribosome contents varied within the normal and decreased values in both the patient groups. The specific radioactivity in 28S rRNA leukemic lymphocytes was significantly lower in groups of patients with low ribosome contents than in those with the normal ones. The data suggest that in the leukemic cells with a high or unaltered activity of ribosome cistrons and low ribosome levels rRNA processing is broken.

[Interrelation between the ribosome content of lymphocytes and hypogammaglobulinemia in chronic B-cell lymphatic leukemia]. / O vzaimosviazi mezhdu kolichestvom ribosom v limfotsitakh i gipogammaglobulinemiei pri B-kletochnom khronicheskom limfoleikoze.

The serum immunoglobulin levels, ribosome contents and protein synthesis rate in peripheral blood lymphocytes have been studied in 35 patients with chronic lymphatic leukemia. It is shown that in patients whose lymphocyte ribosome content corresponds to that of the donors IgG, IgA, IgM levels were on the average normal whereas in the patients with the decreased ribosome content IgG and IgA were 2-2.7 times lower, while the IgM concentration was normal. Hypogammaglobulinemia of IgG, IgA and IgM types in 47% of patients with the normal ribosome contents was 23%, 29% and 23%, respectively, and in 100% of patients with the low ribosome content it was 72%, 83% and 68%, respectively. There was no distinct correlation between the immunoglobulin levels and the protein synthesis rate. The ribosome pattern in the lymphocyte maturation in chronic lymphatic leukemia is discussed.

[Selection of appropriate ribonuclease inhibitors in the study of polysomes in human lymphocytes]. / K voprosu o podbore adekvatnykh ingibitorov ribonukleaz pri izuchenii polisom v limfotsitakh cheloveka.

Polysome profiles are described for lymphocytes of donor peripheral blood treated with various inhibitors of RNAases. The most effective inhibitor was diethyl pyrocarbonate, which enabled to maintain high content of polysomes (up to 70%) in these cells. Ribosomes, treated with diethyl pyrocarbonate, exhibited functional activity in the puromycin test. A natural inhibitor of RNAases from liver tissue inhibited incompletely the enzymatic activity in lymphocytes. Heparin proved to be an inadequate inhibitor of RNAases in all the cells of lymphoid origin (lymphocytes from peripheral blood of donors and of the patients with chronic lympholeukosis, lymphocytes isolated from adenoids and spleen tissue); it induced disaggregation of polysomes and dissociation of ribosomes.

[Effect of cyclophosphane, chlorbutin and degranol on protein biosynthesis in the lymphocytes of donors and chronic lympholeukemia patients in vitro]. / Vliianie tsiklofosfana, khlorbutina i degranola na biosintez belka v limfotsitakh donorov i bol'nykh khronicheskim limfoleikozom in vitro.

The alkylating agents in different concentrations are studied for their effect on protein biosynthesis and the relative content of leu-tRNA. The rate of elongation and termination of polypeptide translation in lymphocytes of donors and of patients with chronic lymphoid leukemia in vitro is studied. The therapeutic doses of cyclophosphamide are shown to inhibit the amino acid incorporation into acid insoluble pool of predominantly leukemic lymphocytes, while chlorbutin and degranol caused the greater suppression of the amino acid incorporation into healthy cells. Chlorbutin, cyclophosphamide and degranol in doses above therapeutic caused approximately 95, 48 and 34% inhibition of the protein synthesis, respectively. The mechanism of action on the translation is different. Only chlorbutin acted directly on the translation rate and also increased leu-tRNA content. Degranol decreased the leu-tRNA content. Cyclophosphamide produces no effect on these processes in vitro.

[Poly(A) RNA distribution and the polysome-monosome ratio in the lymphocytes of healthy persons and chronic lympholeukemia patients]. / Raspredelenie poli(A) RNK i sootnoshenie polisom i monosom v limfotsitakh zdorovykh i bol'nykh khronicheskim limfoleikozom liudei.

Intact polysomes from lymphocytes of donors and patients with chronic lymphocytic leukemia (CLL) were isolated using diethyl pyrocarbonate. It has been shown that only 3/4 of ribisomes are bound to polysomes of a comparatively small size. Poly(A) RNA of the postmitochondrial fraction is largely associated with polysomes. Cyclohexamide treatment results in recruitment of monosomes into polysomes and in a shift of ribosomes in larger polysomes. There are no differences in the polysome content and size, the effect of cyclohexamide treatment and the distribution of the polysomal and non-polysomal poly(A) RNA in normal and CLL lymphocytes. It is suggested that during the transition of resting lymphocytes into the growth state, when protein synthesis increases, the mobilization of mRNP and ribosomes into polysomes is not essential.

[Biochemical heterogeneity of lymphocytes in chronic lympholeukemia based on the analysis of the protein-synthesizing apparatus]. / Biokhimicheskaia neodnorodnost' limfotsitov pri khronicheskom limfoleikoze po dannym analiza beloksinteziruiushchego apparata.

Duration of middle polypeptide synthesis, the rate of protein synthesis, content of ribosomes in lymphocytes were studied in the cells from 9 healthy persons and 28 patients with lympholeukosis. In 14 of the patients examined the content of ribosomes was decreased 2-fold; in the other patients it approached the control values. The rate protein synthesis was distinctly decreased in the patients with low content of ribosomes; in patients with normal content of the ribosomes this rate was either increased or decreased. Acceleration of the middle peptide synthesis was detected in leukemic lymphocytes as compared with controls. The data obtained suggest that lymphocytes exhibited biochemical heterogeneity in chronic lympholeukosis as well as that not only the content of ribosomes but other factors, importance of which in regulation of protein synthesis is discussed, influenced the rate of protein synthesis in lymphocytes of the patients.

[Relative polyribosome content in peripheral blood lymphocytes and lymphoid organs in the human]. / Otnositel'noe soderzhanie poliribosom v limfotsitakh perifericheskoi krovi i limfoidnykh organov cheloveka.

Using ultracentrifugation, relative contents of polysomes and free ribosomes were calculated in human PHA activated or non-activated peripheral lymphocytes, peripheral chronic lymphocytic leukemia lymphocytes, and in spleen and adenoid lymphocytes in the total fraction as well as in that separated into T and B lymphocytes. The polysome amount in B cells was higher (47%) that in T cells (30%). The polysome and free ribosome correlation in lymphocytes of lymphoid organs and in peripheral blood was different to be presumably dependent on population composition of lymphocytes and on their maturation and proliferation. 20-25% of ribosomes in peripheral lymphocytes are found in polysomes; in PHA stimulated leukemic and spleen lymphocytes they reach 50-70%, and in adenoid lymphocytes, -40%. The participation of polysomes, rather than monosomes, in protein synthesis in lymphocytes has been shown irrespective of the polysome--free ribosome correlation in these.

[Comparative study of the lymphocyte protein-synthesizing activity of donors and chronic lympholeukemia patients]. / Sravnitel'noe izuchenie beloksinteziruiushchei aktivnosti limfotsitov donorov i bol'nykh khronicheskim limfoleikozom.

An assay of lymphocyte ribosomes in blood of donors and patients with chronic lymphocytic leukemia established the normal level of ribosomes in 11 cases and half that value in 6 cases of leukemia. The decreased level of ribosomes in cells was largely, registered at the terminal stage of the disease. The 14C-leucine uptake by leukemic lymphocytes was reduced 2-3-fold while the relative level of 14C-leucyl-tRNA in diseased and normal cells was identical. The rate of lymphocyte protein synthesis was lowered in a wide range (2-8-fold), in most of patients (7 out of 8). The rate of protein synthesis (radioactivity of acid insoluble fraction per mg of ribosomes) was half the normal value and constant in leukemic lymphocytes. An important role of ribosome level in protein biosynthesis regulation in the course of disease progression is suggested.

[Comparative study of nuclear RNA in leukocytes of normal individuals and patients with leukemia by the method of molecular DNA-RNA hybridization]. / Sravnitel'noe izuchenie iadernykh RNK leikotsitov cheloveka v norme i pri leikozakh metodom molekuliarnoi DNK-RNK-gibridizatsii.

Spectra of nuclear RNA were studied in leukocytes in healthy donors and in patients with leukoses by DNA-RNA hybridization. Cross competitive hybridization showed that in chronic lymphoid leukosis and acute myeloblastosis new forms of nuclear RNA were observed, which were not found in normal state; in myeloleukosis the nuclear RNA spectra were similar to those of normal granulocytes. The results obtained suggest that universal type of impairments appears to be absent in regulation of transcription in leukoses; it may be related not only to leukosis transformation but depends on variations in cell content of leukocyte populations studied.