[Oncolytic parvoviruses. A new approaches for cancer therapy].

Parvoviruses such as parvovirus H-1 (H-1PV) may selectively infect and lysis cancer cells. The parvoviruses also induce an immune system to eliminate the tumor cells through the formation of anti-cancer immunity. One of the possible mechanisms of antitumor activity is associated with the direct induction of apoptosis by parvoviral proteins NS1 and 11 kDa. Parvovirus-based vectors are promising for gene therapy of oncological diseases and genetic disorders in humans. Parvoviruses were successfully used for the experimental treatment on animal models of human glioma, neuroblastomas, lymphomas, pancreatic carcinoma, carcinomas and breast tumors. ParvOryx is the first oncolytic preparation constructed on the base of H-1PV; its phase I/IIa clinical trials in patients with glioblastoma multiforme are in process.

[The Russian revision of chronic gastritis classification: reproducibility of the pathomorphologic picture].

The estimation of Russian revision reproducibility of the chronic gastritis OLGA system International classification has been spent. The analysis of Russian pathologists-experts work, considered in estimations of an expression level and prevalence of gastric mucosal atrophic changes, as a tumour predictor, for the identification of chronic gastritis stages (0-IV) has been carried out by kappa-statistics. The different levels of experts' consent according to consent criteria calculation have been estimated. The criteria of consent (k) of leading Russian pathologists have been 0.5 (moderate level of the agreement). In the practice of histopathology researchers' classification a consent level has been lower--from 0.27 (satisfactory or tolerable consent level) to 0.42 (moderate or average consent level). A subjective reasons reducing consistency level of pathologists-experts have been discussed.

[Immune response in patients with HCV infection on the south of western Siberia].

UNLABELLED: The aim of the study was to investigate the features of immune response in patients with HCV infection on the South of Western Siberia. MATERIALS: The 130 patients with different genotips of HCV infection were investigated (95 men and 35 women). RESULTS: On the territory of the South of Western Siberia the 1 and 3 genotips of HCV infection were present especially in young men and women and 1b subtype was dominated. In patients with 1 and 3 genotips of HCV infection was occurred the massive inflammatory response with intensive necrotic proinflammatory cytokins damages of the liver tissue and high cirrotic occurring in young age.

[Cell therapy in treatment of trophic ulcers of lower extremities].

The aim of the work was to study the effectiveness of using human embryo fibroblast culture in complex treatment of trophic ulcers of venous etiology in 23 patients with trophic ulcers of lower extremities. The cause of the appearance of ulcers was postthrombophlebitic disease in 16 patients and varicose disease in 7 patients. A control group consisted of 25 patients (postthrombophlebitic disease in 18 patients and varicose disease in 7 patients). The human embryo cell culture grown on the wound cover "Foliderm" was used at the stage of epithelization in the main group, while in the control group the modern alginate, collagen, hydrogel, polyurethane and hydrocolloid covers Suprosorb--Suprosorb A, Suprosorb C, Suprosorb G, Suprosorb F and Suprosorb H were used. Healing of the varicose trophic ulcers in the control group was achieved in 86% of patients, of post-thrombophlebitic--in 78% of patients. The average period of healing was 3.6 and 3.9 months respectively. Healing of trophic ulcers in the main group took place in 100% of patients. The average period of healing was 1.5 week for varicose and 3.2 weeks for postthrombophlebitic ulcers. The cell therapy was shown to be a highly effective method in treatment of venous trophic ulcers.

[Phezam efficacy in patients with chronic cerebral ischemic disease].

The efficacy of combined medication which comprised compounds with nootropic (piracetam) and vasoactive (cinnarisin) effects, was studied in patients with cerebral blood flow insufficiency. The main inclusion criterion was a diagnosis of chronic brain ischemia (CI). The study consisted of two stages: (1) a randomized comparative trial in neurological clinic (60 patients) and (2) estimation of the drug efficacy in routine practice (60 patients). The clinical examination was accompanied by neuropsychological tasks, kinetic tests and ultrasound investigation of brain vessels. At the first stage, a positive neurological and neuropsychological dynamics was found after 8 weeks of phezam treatment. Also a statistically significant positive dynamics was observed for a number of blood flow velocity parameters in the middle brain artery. In routine medical practice, a positive effect of phezam was seen after 2 months of the treatment for all but CI main symptoms and confirmed by the data of kinetic investigation. The patients reported good tolerability and convenience of the drug intake (one capsule instead of two tablets of nootropic and vasoactive drugs).

[Development of microencapsulating measles live vaccine]. / Razrabotka mikrokapsulirovannoi formy zhivoi korevoi vaktsiny.

Designing of non-injection methods of immunization against measles has recently turned into a topical issue. Development of mucosal vaccines ensuring the "entry gate" immunity, which is highly effective in airborne infection, is in the focus of attention. The authors developed a method of microencapsulating the viral particles into the matrix of pH-dependent polymers. Microencapsulated live measles vaccine shaped as 0.6-2.0 microm particles was obtained. The specific activity of measles virus in the drug was 3.36-4.31 log TCD50/0.5 ml. In subcutaneous immunization of guinea pigs with capsules, the best results were obtained in a single administration of vaccine based on ethylcrylate, sodium alginate/ chitosan and sodium slaginate/HMDA. In the intranasal administration of vaccine based on sodium alginate/spermin and sodium alginate/HMDA, there was a need in 2 and 3 stages of immunization.

[The influence of thermoperiod on cucumber growth and development]. / Vliianie termoperioda na rost i razvitie ogurtsa.

We studied the influence of gradient temperature regimes on various parameters of the formation of above-the-ground and underground organs of cucumber plants, such as rate of leaf appearance, rate of growth, duration of growth and length of leaves, and the rate of growth of above-the-ground organs and roots. The plants were grown under the controlled conditions: at different combinations of day and night temperature, illumination 100 Wt/m2, and 12 h photoperiod. The comparison of constant and fluctuating diurnal temperature regimes has shown that in the optimal area for all studied indices, the highest values were recorded at the constant daily temperature (25 degrees C for all growth indices of above-the-ground organs and 20 degrees C for growth of roots), while all gradient regimes either did not affect, or exerted inhibitory effects on the plant. The main acting fluctuating temperatures, that exerted stimulating effects, combined low hardening (15 degrees C) and optimal temperatures (25 degrees C), which was earlier described for animals. The 15/35 and 35/15 degrees C combinations were unambiguously inhibitory, since both temperatures are hardening for the cucumber. A lesser stimulating effect of the developmental rate in a plant, as compared to poikilothermic animals, could be due to a greater autonomy of plant ontogenesis because of autotrophy and, correspondingly, a greater degree of homeostasis. The mechanisms accounting for the reactions to temperature gradients are similar in different groups of ectotherms.

[Course of heart failure in patients with terminal chronic kidney failure on chronic hemodialysis]. / Osobennosti techeniia serdechnoi nedostatochnosti u patsientov s terminal'noi khronicheskoi pochechnoi nedostatochnost'iu, poluchaiushchikh lechenie khronicheskim gemodializom.

AIM: To evaluate peculiarities of the course of cardiac failure (CF) in initiation of hemodialysis therapy (HT) in patients with terminal chronic renal failure (CRF) and 24 months after HT; to elucidate CF causes late in HT. MATERIAL AND METHODS: Cardiohemodynamics was studied in 152 patients with terminal CRF during 2 years of HT. RESULTS: At initiation of HT, cardiohemodynamics was characterized by hyperkinetic syndrome, high total peripheral resistance, weak left ventricular systolic function, diastolic dysfunction. Chronic HT for 2 years led to attenuation of hyperkinetic syndrome, improvement of left ventricular systolic function, highly increased bypass blood flow along the arteriovenous fistula. The correction of the blood flow along the arteriovenous fistula arrested manifestations of CF in all the patients. CONCLUSION: Drugs with positive inotropic action are contraindicated in patients with terminal CRF on chronic hemodialysis having CF. Regular measurements of blood flow along the arteriovenous fistula are recommended for early detection and correction of increased bypass blood flow.

[Extrapulmonary tuberculosis in children in Russia (epidemiology, clinical forms and their study)]. / Vnelegochnye formy tuberkuleza u detei v Rossii (épidemiologiia, klinicheskie formy i ikh nabliudenie).

The paper provides the data of comprehensive epidemiological and clinical studies made at the Clinic of Tuberculosis in Children and Adolescents. Research Institute of Phthisiopulmonology, I. M. Sechenov Moscow Medical Academy, Ministry of Health of the Russian Federation, in the past 15 years and dedicated to extrapulmonary tuberculosis in children and adolescents with tuberculosis. Due to the specific features of manifestations of extrapulmonary tuberculosis in childhood and adolescence and to the existing system of prevention and early detection of the disease in children and teenagers in the Russian Federation, the number of children with severe generalized extrapulmonary tuberculosis is not on the rise despite that the epidemiological situation has aggravated in the past 15 years. The clinical features of most common forms of extrapulmonary tuberculosis are shown and measures for their monitoring proposed.

The monoclonal antibody TER-119 recognizes a molecule associated with glycophorin A and specifically marks the late stages of murine erythroid lineage.

The antigen specificity of a rat monoclonal antibody TER-119 was investigated. In adult mice, TER-119 reacted with mature erythrocytes, 20-25% of bone marrow cells and 2-3% of spleen cells but not with thymocytes nor lymph node cells. In fetal haematopoietic tissues, 30-40% of d 10 yolk sac cells, 80-90% of d 14 fetal liver cells and 40-50% of newborn liver cells were reactive with TER-119. TER-119+ cells in adult bone marrow expressed significant levels of CD45 but not myeloid (Mac-1, Gr-1) or B-cell (B220) markers. Morphological examination and haematopoietic colony-forming assays for isolated TER-119+ cells revealed that TER-119 reacts with erythroid cells at differentiation stages from early proerythroblast to mature erythrocyte, but not with cells showing typical erythroid blast-forming unit (BFU-E) and erythroid colony-forming unit (CFU-E) activities. Erythroleukaemia cell lines do not express the TER-119 antigen even after stimulation with dimethylsulphoxide. TER-119 immunoprecipitated protein bands with molecular masses of 110 kDa, 60 kDa, 52 kDa and 32 kDa from erythrocyte membrane, whereas only a 52-kDa band was detected by TER-119 in Western blot analysis. Further molecular and cellular analyses indicated that the TER-119 antigen is a molecule associated with cell-surface glycophorin A but not with glycophorin A itself.

Mobilization of hematopoietic primitive and committed progenitor cells into blood in mice by anti-vascular adhesion molecule-1 antibody alone or in combination with granulocyte colony-stimulating factor.

OBJECTIVE: One of the mechanisms for mobilization of hematopoietic stem cells and progenitor cells is alternation of adhesion molecules. We investigated the mobilization of hematopoietic progenitor cells in blood by administration of anti-vascular cell adhesion molecule (VCAM)-1 antibody (Ab) in mice. MATERIALS AND METHODS: Twelve- to 14-week old C57BL/6J mice were injected intravenously with anti-VCAM-1 Ab and anti-very late antigen (VLA)-4 Ab at a dose of 5 mg/kg for 2 days. RESULTS: The number of colony-forming cells (CFCs) in blood was increased 11.4-fold after anti-VCAM-1 Ab treatment, but the number of CFCs was not increased after treatment with anti-VLA-4 Ab. The number of colony-forming unit spleen (CFU-S) also was increased 21.6-fold in the peripheral blood by administration of anti-VCAM-1 Ab. The number of CFCs and CFU-S in the bone marrow of mice treated with anti-VCAM-1 Ab was decreased and that in the spleen also was decreased. On administration of recombinant human granulocyte colony-stimulating factor (125 microg/kg twice daily) with anti-VCAM-1 Ab, the numbers of CFCs and CFU-S were increased 141.8-fold and 439-fold, respectively. CONCLUSIONS: These observations demonstrated that administration of anti-VCAM-1 Ab induced mobilization of hematopoietic progenitor cells into blood from bone marrow and spleen and that granulocyte colony-stimulating factor has synergistic effects on anti-VCAM-1 Ab-induced mobilization.

[Preparation of P52 recombinant antigenic protein from human cytomegalovirus (HCMV)]. / Poluchenie rekombinantnogo antigena belka P52 tsitomegalovirusa cheloveka (HCMV).

Analysis of published reports helped us single out the most potent antigens among HCMV proteins: phosphoproteins pp150(UL32) and p52(UL44). Theoretical computer analysis of p52 epitopes showed the main antigenic determinants not cross-reacting with antigens of other viruses. Virus-containing (strain AD169) material was obtained and genome DNA was isolated. Amplification of a site of gene UL44 coding for unique determinants detected a PCR fragment of required electrophoretic mobility. The fragment was cloned in vector pLBE. The specificity of cloning was confirmed by restriction analysis of theoretical sites. Nucleotide sequence of cloned fragment of UL44 gene was studied by Maxam-Gilbert's method. Cloning in expressing bacterial vectors helped obtain HCMV recombinant protein p52 in the pure form and fused with beta-galactosidase. Enzyme immunoassay with HCMV-positive and negative donor sera and ABBOTT HCMV sera showed that recombinant p52 increased the sensitivity and specificity of a previously obtained recombinant pp150 as an antigen to HCMV-IgG and HCMV-IgM. The sensitivity and specificity is 100% with 98-99% reliability.

Selective expression and function of granzyme D in lymphohematopoietic stromal cells.

Granzymes are a family of serine proteases exclusively detected in the granules of cytotoxic lymphocytes, and in mice at least eight granzymes, A to G and K, have been identified. Except for granzymes A and B, which activate the apoptotic pathway, little is known about the exact functions of the other granzymes. We have found that the granzyme D transcript is selectively expressed in functional hematopoietic stromal lines as well as primary stromal cells. Stromal lines supported growth of a pre-T lymphoma clone BTK at an efficiency proportional to the expression level of granzyme D, while a stromal line lacking granzyme D failed to do so. When the defective stromal line was transfected with granzyme D cDNA, it could efficiently support the growth of BTK cells. The results indicate that granzyme D expressed in the stromal cells plays an important role in stromal cell-lymphocyte interaction.

Expression of VCAM-1 in lymphocytes during the process of apoptosis.

The administration of corticosteroids induced apoptosis of thymocytes in vivo. Among various adhesion molecules examined, vascular cell adhesion molecule-1 (VCAM-1, CD106) was shown to be strongly expressed in these apoptotic cells. Flow cytometric analysis also showed the expression of VCAM-1 in apoptotic thymocytes. An RT-PCR study demonstrated the expression of VCAM-1 mRNA in thymocytes. Splenic lymphocytes and other lymphoid cell lines also expressed VCAM-1 during the process of apoptosis. VCAM-1 mRNA expression was also observed in RT-PCR performed on these cell lines.

[Elaboration of live measles vaccine technology on human embryo lung diploid cell culture L-68]. / Razrabotka tekhnologii proizvodstva zhivoi korevoi vaktsiny na osnove kul'tury kletok émbriona cheloveka L-68.

Measles predominates among childhood droplet infections in many countries. Immunization of all human beings sensitive to this infection is the only radical measure in controlling measles. The quality of a vaccine is primarily determined by the properties of the virus strains and cell cultures and technology of production. Now live measles vaccine is produced in or country on the basis of fibroblasts from Japanese quail embryo. The production of live measles vaccine in the primary cell cultures has a number of drawbacks caused by the nonstandard pattern of the substrate and the probability of contamination. The use the certified human diploid cells deposited in liquid nitrogen in sufficient quantities is promising. The authors have elaborated a new technology of live measles vaccine production by using the Leningrad-16 virus strain on the basis of attested L-68 diploid cell culture from the human fetal lung. Experimental batches of vaccine were obtained and attested in accordance with the present requirements for immunobiological products.

Enhancement of activation-induced cell death by fibronectin in murine CD4+ CD8+ thymocytes.

Development of T cells in the thymus is achieved through the interactions of thymocytes with their microenvironments. This study focused on the function of fibronectin (FN), a major extracellular matrix molecule in the thymus, in the cell death induced by activation via the T-cell antigen receptor. FN alone did not increase cell death in murine thymocytes above the baseline level, but it significantly enhanced the cell death induced by fixed anti-CD3 monoclonal antibody (mAb), especially when a high concentration of anti-CD3 mAb was used. DNA fragmentation increased in parallel with cell death, indicating that cell death was a result of the apoptosis. Fluorescence-activated cell sorter (FACS) analysis revealed that the activation-induced cell death (AICD) caused by anti-CD3 mAb alone, or by a combination of anti-CD3 mAb and FN, occurred selectively in CD4+ CD8+ thymocytes. Very late activation antigen (VLA)-4 and VLA-5 are two major ligands to FN on thymocytes. The expression of both ligands was investigated at different stages of thymocyte development. VLA-4 was predominantly expressed at the CD4- CD8- stage, and thereafter the expression was reduced, whereas VLA-5 was constantly expressed during maturation. Furthermore, the enhancing effect by FN was inhibited in the presence of the Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP) peptide but not in the presence of the connecting segment-1 (CS-1) peptide, suggesting that enhancement of AICD observed in CD4+ CD8+ thymocytes is mediated through VLA-5.

[Features of adult T-cell leukemia virus reproduction in B-cell lines and it's interaction with the Epstein-Barr herpesvirus]. / Osobennosti reproduktsii virusa T-kletochnogo leikoza vzroslykh v B-kletochnykh liniiakh i ego vzaimodeistviia s gerpesvirusom Epshteina-Barr.

Interaction between human T-cell leukemia virus (HTLV) with B cells and Epstein-Barr virus (EBV) was studied by immunoblotting, immunofluorescence, virus isolation in permissive T-cell cultures, and polymerase chain reaction (PCR). HTLV-1 in vitro infects the B-cell cultures containing EBV but not EBV-negative cell lines. Productive infection of EBV+ B cells was associated with syncytium formation which led to the elimination of HTLV-1 producing cells. However, the remaining B-cell population contained gag, pol, and pX--the "silent" provirus sequences. HTLV-1 infection of B cells altered the expression of some latent proteins of EBV (EBNA-1, EBNA-2, EBNA-5, and LMP). The changes were represented by increase of molecular weight and/or appearance of additional proteins and were individual for each cell line. Alteration of EBV protein expression may change the functional activity of these proteins, but this hypothesis is to be tested.

Modulation of cell surface lectin receptors on K562 human erythroleukemia cells induced by transfection with annexin IV cDNA.

Annexin IV was found to be highly expressed in various human adenocarcinoma cell lines, but not in an erythroleukemia cell line, K562. We investigated the effects of transfection of human annexin IV cDNA into K562 cells on cell surface lectin receptors. cDNA transfectants were found to be more sensitive to cytotoxic lectins such as Ricinus communis agglutinin and wheat germ agglutinin than mock transfectants. The results of flow cytometric analyses with various lectins showed that the transfectants expressed more sugar chains which bind to Ulex europaeus agglutinin I and Maackia amurensis mitogen than mock transfectants. These results suggest that transfection of annexin IV cDNA increases the expression of alpha-2,3-sialylated and/or fucosylated sugar chains on the surface.

Characterization of human p33/41 (annexin IV), a Ca2+ dependent carbohydrate-binding protein with monoclonal anti-annexin IV antibodies, AS11 and AS17.

p33/41 (annexin IV) is a member of the family of Ca(2+)-dependent phospholipid binding proteins known as annexins. We previously described that bovine kidney p33/41 (annexin IV) has Ca(2+)-dependent carbohydrate binding activity. In this study, we purified human p33/41 (annexin IV) from the HT29, human colon adenocarcinoma cell line, as well as the bovine kidney annexin by affinity chromatography. Then, we prepared recombinant human p33/41 (annexin IV) expressed in Escherichia coli. The apparent size and the Ca(2+)-dependent carbohydrate binding properties of purified recombinant p33/41 (annexin IV) were indistinguishable from those of the bovine kidney protein. We also performed inhibition assays of carbohydrate binding and of phosphatidylserine/phosphatidylcholine liposome binding of recombinant p33/41 (annexin IV) with anti-p33/41 monoclonal antibodies (AS11 and AS17). We determined the epitopes recognized by the monoclonal antibodies by Western blot analysis using deleted-recombinant p33/41 (annexin IV). The monoclonal antibodies recognized domain 1 and/or 2 of p33/41 (annexin IV). The results of the inhibition assays and the determination of the epitope showed that a carbohydrate binding site is located at domains 3 and 4 of p33/41 (annexin IV) and on the cell surface.