[The distribution of iodine-125 labeled alpha-fetoprotein in the animal organism and its accumulation in the tumor].

The distribution of iodine-125 labeled human alpha-fetoprotein in mice was studied after its intravenous injection. The maximal accumulation of alpha-fetoprotein in different tissues and organs of animals was observed mainly 5 hours after injection. Then the protein was gradually eliminated from the body. In the liver, intestine and blood of intact animals 125I-alpha-fetoprotein persists for at least three days. Accumulation of alpha-fetoprotein in various tissues and organs may determine the different biological effects of this protein. In the mice with transplanted lymphatic leukemia cells P388 the high level of alpha-fetoprotein accumulation was detected in the tumor tissue, reaching 6% of the injected amount per 1 g of tissue. This allows considering the radionuclide-labeled alpha-fetoprotein as a promising medical radionuclide marker for the radiological detection of malignant tumors.

[Regulation of human dendrite cells' physiological functions with the recombinant heat shock protein Hsp70].

Reference literature review. The dendrite cells (DC) are the most important antigen-representing cells of the organism. They are the target for various vaccines, and on the basis of the DC cellular anti-tumour and anti-viral vaccines (DC vaccines) have been developed. At the same time, the DC can be a convenient model for studying activity and action mechanisms of different immune-therapeutic preparations. One of the aspects of the DC use optimization for induction of antigen-specific immune response might involve use of the heat shock proteins (Hsp), the Hsp70 in particular. This protein can be used for administration of protein antigens into the DC and for regulation of the DC activity. Knowledge of the DC physiology and specifics of interrelationship among the Hsp70 and its complexes with the DC antigens of different differentiation degree is an important aspect for implementation of these ideas. Human Hsp70 has been shown both to bring antigens to the DC and to regulate activity of the DC as well as to optimize induction of antigen-specific cellular immune response.

[Inactivation and sensitization of the tumor cells by the gene Bax transfection].

After the transfection of the gene Bax into the cultured tumor cells of human ovary adenocarcinoma SKOV3 and uterus carcinoma HeLa in vitro the high sensitivity of the cells SKOV3 to the protein Bax produced after the gene Bax transfection was found. The sensitivity of the cells HeLa to the gene Bax transfection was much smaller. The hyperexpression of gene Bax and hypersensitivity to doxorubicin were seen in HeLa cells received as a result of the gene Bax transfection and subsequent selection. All cells of the line SKOV3 with the increased expression of the transfected gene Bax died. In the cell line SKOV3 the mutation in a gene Bax was found which has a genotype G7/G9 against a native type of a gene Bax--G8/G8. It was concluded that the found in the exone 3 of the gene Bax mutation G7/G9 in cells SKOV3 results in an inactivation of proapoptotic activity of the protein Bax.

[The role of Fas/FasL system in induction of hepatocyte apoptosis in chronic viral hepatitides]. / Rol' sistemy Fas/FasL v induktsii apoptoza gepatotsitov pri khronicheskikh virusnykh gepatitakh.

The aim of the study was assessment of hepatocyte apoptosis depending on expression of Fas and FasL proteins by various liver cells in patients with chronic viral hepatitis B (CVHB) or chronic viral hepatitis C (CVHC). The symptoms of hepatocyte apoptosis were observed in 3 of 12 patients with CVHB and in 9 of 14 patients with CVHC, the proportion of apoptotic cells being 12-65%. Hepatocytes of healthy people and patients with hepatitis B or C express Fas protein in the cytoplasm diffusely, as granules or on cell membrane. In health, hepatocytes do not express FasL, but in CVH they do. The highest apoptosis was observed in Fas protein location as granules in cytoplasm or in their preferable location on the cell membrane. The severity of hepatocyte apoptosis in CVH directly correlated with FasL expression by the cells of the lymphoid-histiocytic infiltrate in the liver and inversely correlated with FasL expression by hepatocytes. Thus, a great part of hepatocytes in CVH are killed by the virus; Fas/FasL interaction is leading in damage to hepatocytes in CVH.

[Deletion mutants of human granulocyte-macrophage colony-stimulating factor]. / Deletsionnye mutanty granulotsitarno-makrofagal'nogo koloniestimuliruiushchego faktora cheloveka.

To study the structure-function relationship of the human granulocyte-macrophage colony-stimulating factor (GM-CSF), genes were constructed that encode its three deletion mutants: D1, a mutant with the deletion of six amino acid residues (37-42) some of which are a part of a beta-structural region; D2, a mutant with the deletion of the unstructured six-aa sequence of a loop (45-50); and D3, a mutant with the deletion of 14 aa residues (37-50) corresponding to the A-B loop and encoded by the second exon of the gmcsf gene. The expression products of these genes in E. coli were accumulated in a fraction of insoluble proteins. The secondary structures of the mutant proteins were similar to that of the full-size GM-CSF, but the biological activity of the deletion mutants was 130 times lower than that of the GM-CSF: they stimulated the proliferation of the TF-1 cell line at 3 ng/ml concentration. The resulting proteins displayed antagonistic properties toward the full-size GM-CSF, with the inhibition degree of its colony-stimulating activity being 27%. A decrease in the mutant activity in the row D2 > D1 > D3 implies the importance of the conserved hydrophobic residues involved in the formation of the beta-structure for the formation of the GM-CSF functional conformation.

[Clinical evaluation of the structural characteristics of peripheral blood leukocyte DNA in patients with viral hepatitis B]. / Klinicheskaia otsenka pokazatelei struktury DNK leikotsitov perifericheskoi krovi bol'nykh virusnym gepatitom B.

The DNA structure of peripheral blood leukocytes in healthy donors and in viral hepatitis B patients was studied by the rate of the alkaline denaturation of DNA in cell lysates. An increased rate of the DNA alkaline denaturation of cell lysates was established, which was indicative of the damages in their DNA. The most pronounced damages of DNA were found in granulocytes of patients with highly active chronic hepatitis and hepatic cirrhosis, especially in cases of simultaneous delta-virus infection. The damage of leukocyte DNA reflected probably the accumulation of cells, committed to apoptosis, in peripheral blood. Apoptosis may be induced by cytotoxic lymphocytes recognizing HBV-infected cells.

[The structure of the leukocyte DNA in leukemia patients during chemotherapy]. / Struktura DNK leikotsitov bol'nykh leikozom v protsesse khimioterapii.

The authors studied the degree of DNA damage in in vitro cultured human peripheral lymphocytes (PL) and Jurcat's human T-cell lymphoma cells exposed to a stabilized 4 OH-cyclophosphan-mamophosphatide (MA) derivative, as well as in the leukocytes from patients with leukemia who were treated with cyclophosphan. There was an increase in alkaline DNA denaturation rate of LP lysates and T-cell lymphoma cells, which was in proportion to MA concentrations, and a higher sensitivity of LP to the genotoxic effect of MA given in doses of 5-10 micrograms/ml than that of Jurcat's cells, as well as high peripheral lymphocyte and neutrophil DNA damages in patients with leukemia during chemotherapy. The authors consider that the accumulation of single-strand breaks and alkaline-labile sites, which was recorded from the increase in alkaline DNA denaturation rate of cell lysates, is a highly sensitive test for detecting DNA damages in resting and slowly proliferating cells and can be useful in revealing and evaluating the severity of human genotoxic effects.

[Cyclic AMP content, protein kinase activity, and DNA structure in resting and proliferating peripheral blood lymphocytes and in human T-lymphoma cells]. / Soderzhanie cAMP, aktivnost' proteinkinaz i struktura DNK v pokoiashchikhsia i proliferiruiushchikh limfotsitakh perifericheskoi krovi i v kletkakh T-limfomy cheloveka.

Alterations of DNA structure, of cAMP content, of cAMP-dependent histokinases (HK) activity and cAMP-independent casein kinases (CK) activity were studied during transformation of resting cells to proliferation. These patterns were studied in human lymphocytes from peripheral blood immediately after their isolation and after cultivation within 3 days in presence of concanavalin A (ConA) or without the mitogen as well as in cultivated cells of human T-lymphoma Jurkat. Increase in content of alkaline labile sites in DNA, in activity of CK as well as distinct increase in content of cAMP were detected within the first 18 hrs of lymphocytes cultivation both in presence of ConA or without it. Early steps of cell transformation from G0 phase to G1 may be related to these alterations observed. Only slight increase in content of the alkaline labile sites in DNA and decrease in cAMP content were found in both these cell cultures. Activity of CK in the lymphocytes culture not containing ConA was decreased down to initial level, while in presence of the mitogen the enzymatic activity was increased and within 3 days it reached the level of CK activity in Jurkat cells. The rate of CK relative activity, calculated as CK/cAMP or CK/HK/cAMP ratios for each cell preparation, correlated with DNA biosynthesis rate measured by 3H-thymidine incorporation. The data obtained suggest that these patterns, used in differential diagnosis of human large intestine and gastric tumors, demonstrated also the intensity of tissue proliferation.

[Effects of lymphokines on DNA structure of in vitro cultured human lymphocytes. Connection with cAMP and oligoadenylate synthetase systems]. / Vliianie limfokinov na strukturu DNK kul'tiviruemykh in vitro limfotsitov cheloveka, sviaz' s sistemoi tsAMF i oligoA.

We investigated the influence of recombinant interferons (INF) alpha 2 and gamma and interleukin (IL) 2 and natural purified IL 1 on the activity of oligoadenylate synthetase proliferation level and the DNA structure of cultured in vitro human peripheral blood lymphocytes. It was shown that the proliferation of mitogen-stimulated lymphocytes increased in the presence of IL 1, IL 2 and INF gamma, but there was no proliferation in the presence of INF alpha 2. Oligoadenylate synthetase activity was increased after 18 hours incubation in the presence of all these lymphokines, but after 48 and 72 hours it was increased only in the presence of INF alpha 2 or ConA or INF alpha 2 with ConA together. INF alpha 2, INF gamma and IL 1 stabilized the DNA structure of intact and mitogen-stimulated lymphocytes. cAMP and oligoA synthetase are the specific second messengers of the interferon system and they stabilized the lymphocytes DNA structure too. Cultivation of lymphocytes in the presence of RNA and protein biosynthesis inhibitors--actinomycin D and cycloheximide was followed by accumulation of alkali-labile sites in their DNA. That means that some short lived proteins are needed for the stabilization of native DNA structure.

[An analysis of DNA secondary structure during lymphocyte activation by mitogens]. / Analiz vtorichnoi struktury DNK pri aktivatsii limfotsitov mitogenami.

The DNA structure of human peripheral blood lymphocytes was studied following activation by concanavalin A in vitro. DNA strand breaks were measured by the fluorometric method after DNA alkaline denaturation in cell lysates. Stimulation of lymphocytes proliferation was controlled by 3H-thymidine and 3H-uridine incorporation. It was shown that 1-3 hours after the addition of mitogens the quantity of alkali-labile sites in DNA increased. Cultivation of lymphocytes in vitro without mitogens was accompanied by gradual accumulation of alkali-labile sites in DNA and in 18-72 hours the DNA structure of lymphocytes in the culture was the same. It is possible that the rapid disturbances in the DNA secondary structure of lymphocytes following activation by mitogens is determined by the transmembrane signal transduction and is the one of the early events in genome activation.

[Study of the DNA structure of peripheral blood leukocytes, proliferating lymphoid and tumor cells according to the rate of alkaline denaturation of DNA lysates]. / Izuchenie struktury DNK leikotsitov perifericheskoi krovi, proliferiruiushchikh limfoidnykh i opukholevykh kletok po skorosti shchelochnoi denaturatsii DNK lizatov.

Direct fluorometry was used to identify quantitative differences in the DNA structure of human peripheral blood leukocytes and proliferating lymphoid cells. The differences should be taken into account in the study of varied damaging actions.

[Decreased capacity of the peripheral blood lymphocytes for excisional DNA repair in disseminated sclerosis]. / Snizhenie sposobnosti limfotsitov perifericheskoi krovi k ékstsizionnoi reparatsii DNK pri rasseiannom skleroze.

In patients with multiple sclerosis the reduction of DNA excision reparation capacity of peripheral blood lymphocytes was found to correlate with the disease severity but not with its duration. These changes and the increase in the DNA reparative synthesis along with the lymphocytes DNA structural changes are considered as the evidence of impaired lymphocytes genome stability in this ailment. Their possible role in the development of immunodeficiency in multiple sclerosis is discussed.