The Synthesis and Transfection Activity of Disulfide Polycationic Amphiphiles.

Some new polycationic amphiphiles containing a disulfide group were synthesized. Cationic liposomes formed from the compounds synthesized and a helper lipid 1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine were not toxic for HEK293 and HeLa cells and were highly effective when delivering a fluorescently labeled oligodeoxyribonucleotide. The efficacy of plasmid DNA delivery depended on the cell line and the amphiphile structure, liposomes based on tetracationic amphiphiles being the most effective transfectants. These liposomes can be used for in vitro transfection of eukaryotic cells as well as for further in vivo biological studies.

[Synthesis of amino analogues of cholic acid].

Amino analogues of cholic acid were synthesized by reduction of oximes with combination of titanium(III) chloride and sodium cyanoborohydride.

Synthesis and transfection activity of novel galactosylated polycationic lipid.

In this study, we synthesized a new galactosylated cationic lipid and investigated its biological activity. The structure of lipid combines both spermine residue for DNA compaction and galactose moiety for the improvement of aggregation behavior of lipoplexes. Lipid was low toxic for different mammalian cells, and was able both to compact plasmid DNA and to mediate cellular accumulation of various nucleic acids (ODN, pDNA and siRNA) exhibiting biological activity (transgene expression, gene silencing).

[Synthesis of positively charged galactosurfactants].

An approach to synthesis of cationic carbohydrate surfactants with potential antimicrobial or transfected activities is described.

[Synthesis of cationic lipid transfection agents with O,O- or N,O-acetal bonds].

The synthesis of cationic lipids on the basis of 1,2-di-O-tetradecylglycerol is described. The cationic groups represented by the residues of nitrogen-containing heterocyclic bases are attached to the diglyceride backbone through O,O- and N,O-acetal bonds. The resulting lipids can be used as nonviral gene delivery systems to eukaryotic cells.

[Positively charged acyl derivatives of carbohydrates as promising transfection agents].

A convenient approach to the synthesis of mono- and polycationic glycolipid amphiphiles is suggested. The compounds obtained can be used for study of the structure-activity relationship and determination of the effect of hydrophobic and cationic domains on transfection efficiency.

[The synthesis of a glucosyldiglyceride for constructing pH-sensitive liposomes]. / Sintez gliukozildiglitserida dlia pH-chuvstvitel'nykh liposom.

A glucosyldiglyceride containing residues of 11-aminoundecanoic acid was synthesized for constructing pH-sensitive liposomes.

[Transfection of eucaryotic cells using cytochrome CYP450 2B6 gene: sensitivity to cyclophosphamide]. / Transfektsiia éukarioticheskikh kletok genom tsitokhroma CYP450 2B6: poiavlenie chuvstvitel'nosti k tsiklofosfamidu.

Sensitivity of 293 human epithelial kidney cells transfected by human cytochrome CYP450 gene to cyclophosphamide was investigated. Transfection was carried out by plasmid DNA containing CYP2B6 gene complexed with cationic liposomes. Liposomes were prepared from mixture of cationic lipids and cholesterol at different molar ratios. Experimental protocol included the following steps: transfection of epithelial kidney cells by complexes of plasmid DNA-cationic liposomes, clone selection in the medium with antibiotic Geneticin G418, selected clone harvesting and their treatment by cyclophosphamide as following cytotoxicity evaluation. It was shown that addition of 0.25 mM of cyclophosphamide resulted in death of 40-45% transfected cell population.

[Synthesis of cationic glucosyldiglycerides]. / Sintez kationnykh gliukozyldiglitseridov.

A series of glucosyl dialkylglycerols with the pyridinium, N-morpholinium, and N-methylimidazolium polar heads were synthesized by quaternization of the corresponding bases with the acetylated glucosyl diglyceride 6-O-methane- and 6-O-toluenesulfonates. The resulting compounds were designed for the use in gene delivery systems.

[Assay of adenine nucleotide level in platelet storage pool for evaluation of the course of hemoblastosis]. / Issledovanie urovnia adeninnukleotidov pula khraneniia trombotsitov dlia otsenki techeniia gemoblastozov.

The content of adenine nucleotides (ATP, ADP) was studied in dense granules of platelets in hemoblastosis to estimate the character of the pathological process course. Varying biochemical defects were observed at the levels of ATP and ADP depending on the severity of the pathological process in chronic myeloid leukemia, acute lymphoblastic leukemia, acute myeloblastic and acute myelo-monoblastic leukemias. Adenine nucleotide values can be used for the diagnosis of varying stages of the above diseases, for the evaluation of anomalous platelets and characterization of the adequacy of the bone marrow hemopoiesis.

[Importance of studying the level of platelet adenine nucleotide levels in various phases of chronic myeloleukoses]. / Znachenie issledovaniia urovniia adeninnukleotidov trombotsitov v raznykh fazakh zabolevaniia khronicheskim mieloleikozom.

Content of adenine nucleotides was studied in various compartments of blood platelets obtained from donors and patients with chronic myeloleukosis at various steps of the disease. Dissimilar biochemical defect was found in content of ATP and ADP, which varied in thrombocyte storage granules depending on severity of the disease. The data on adenine nucleotides content may be used for diagnosis of various steps of chronic myeloleukosis, for detection of thrombocytes anomaly and for evaluation of the state of medullary hemopoiesis.

[The use of a panel of monoclonal antibodies for immunologic prognosis of recurrence of acute lymphoblastic leukemia]. / Sozdanie paneli monoklonal'nykh antitel dlia immunologicheskogo prognozirovaniia razvitiia retsidivov ostrogo limfoblastnogo leikoza.

A method for early immunological prognosis of acute lymphoblastic leukemia (ALL) relapses has been presented. It is based on the dynamic phenotyping of lymphoid blood cells from the patients during remission by the indirect immunofluorescence test using a specially composed panel with monoclonal antibodies (MCA) to differentiation antigens of early lymphopoiesis stages. The panel composed for monitoring the population of leukemic cells during remission, contained 6 MCA selected from 40 MCA samples with the known specificity, basing on the principle of the highest reactivity with lymphoblasts from patients, and minimal positive reactions with lymphocytes from normal subjects. Blood mononuclear cells from 10 patients with ALL were subjected to immunophenotyping in the panel with MCA during remission, before each course of polychemotherapy. A retrospective analysis of the immunophenotyping data has shown that populations of lymphoid cells carrying antigens of early differentiation stages in amounts significantly exceeding the limits of the standard values (M +/- 3 delta) appear among blood lymphocytes, practically, in all the patients, 1-4 months before the relapse of ALL has been proved.

[The prognostic significance of parameters of bone marrow blast cell proliferation in patients with acute leukemia]. / Prognosticheskoe znachenie parametrov proliferatsii blastnykh kletok kostnogo mozga u bol'nykh ostrym leikozom.

Percentage of bone marrow blasts in S-phase (S%) in acute leukemia patients was evaluated by 3H-thymidine autoradiography or Feulgen cytophotometry at the initial acute period before chemotherapy. In acute lymphoid leukemia (ALL) patients who showed complete remission (CR) S% was 8.6 +/- 1.8% versus 2.1 +/- 0.4% in cases without CR (p less than 0.001). In acute non-lymphoid leukemia (ANLL) the corresponding values were 11.4 +/- 1.4% and 8.3 +/- 1.8%, p greater than 0.05. A negative correlation was observed between S% and the duration of CR in ALL: S% was 13.0 +/- 0.7% in CR, more than 12 months, and 6.9 +/- 1.1% in CR less than 12 months (p less than 0.01). The duration of life also correlated negatively with S%. In ALL S% was 9.5 +/- 1.5% for patients with survival less than 12 months versus 6.2 +/- 0.6% for patients with survival more than 12 months (p less than 0.05). In ANLL the difference (10.3 +/- 1.6% and 6.8 +/- 1.2, respectively) was not significant (p greater than 0.05). Thus in ALL the correlations between S% and the probability to achieve CR is positive, while the correlations between S% and survival and remission duration are negative. In ANLL these correlations are statistically insignificant.