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Biomedical subjects

A I Poletaev

Publications and source records attributed to A I Poletaev.

7 recordsLinked to original sources

Drop-delay measurement using enzyme-coated particles.

A simple technique is suggested for the measurement of drop delay for flow sorting. While the flow cytometer was set to sort a fixed number of particles, the drop-delay setting was changed step by step, and at each step the HRP-coupled particles were sorted into a well of an immunoassay strip. Then the HRP activity of the sorted samples was revealed by routine methods. The maximum level of the enzyme activity shows the proper drop-delay setting. Determination of the drop-delay setting takes only a few minutes. The technique is independent of the type of flow cytometer and does not require any additional equipment.

Animals

[Conformation study of cyclic adenosine-3',5'-monophosphate and some of its derivatives by means of circular dichroism].

Circular dichroism spectra of adenosine and cyclic adenosine-3',5'-monophosphate (cAMP) and their derivatives, having different substituents in 8-position of heterocycle, are studied, cAMP is suggested to have preferable anti-conformation in the solution, while its derivatives with substituents in 8-position of purine base are preferable in sin-conformation. An exception is 8-(beta aminoethylamine-)cAMP, which has an anti-conformation within pH range from 4.5 to 9.5. This is probably due to the formation of intra-molecular ionic bond between cyclophosphate group and aliphatic amino group of 8-position substituent.

Chemical Phenomena

[Circular dichroism of DNA--dye complexes. II. Anisotropy of the long-wave circular dichroism effect and structure of the complex].

Anisotropy of torsional strength of the splitted electronic transition in the case of chromophore-chromophore interaction of dye molecules situated on the helical matrix was considered theoretically and as analytical expression for the value Rperpendicular/Rparallel was obtained. These theoretical results were compared with the experimental data obtained with DNA-proflavine, DNA-pyronine and DNA-acridine orange complexes oriented in multicappilar flow-cell. Studies of the optical effects (optical density and CD changes) due to orientation of these complexes showed that the acridine chromophores are not perpendicular with respect to the DNA axis (alpha D = 19--22 degrees). The DNA base pairs in complexes as assumed also are not perpendicular to the DNA axis, the inclination angle of their transition moments (for the band near 260 nm) being bigger than that of dye chromophores (24 degrees). These results indicate that under experimental conditions used by us no intercalation can be observed.

Acridines

[Secondary structure of RNA of influenza virus in free form and in ribonucleoprotein].

Large amounts of RNA and RNP isolated from influenza virus were obtained. This has allowed us to undertake detailed physical studies of the secondary structure of RNA of influenza virus in free form and in RNP. Analysis of CD spectrum and the hypochromic effect after thermal denaturation of RNA indicated that RNA in free form contains 58--62% double-stranded regions. By comparative studies of the secondary structure of RNA in RNP, it was estimated that 12--14% of the RNA exists in double-stranded form.

Hot Temperature

[Formation kinetics of the AT-specific complex: DNA-distamycin A].

The kinetics of DNA-distamycin A complex formation and dissociation was studied by means of the stop-flow method. It has been found that the complex formation has at least five steps, the formation of AT-specific complex is limited by the dissociation of less specific intermediate complexes, then follows the statistical rearrangement of the ligand molecules and "fixation" of the specific complex with the longest lifetime and the biggest number of the additional H-bounds with the matrix. The energy of a single specific H-bond between the ligand molecule and AT-pair is equal to--1.05 kcal/mol.

Adenine

[Structure of DNA complexes with regular polypeptides].

The conformation of some regular polypeptides: (Lys-Ala)50, (Lys-Ala2)37, (Lys-Ala2)26, (Lys-Ala3)18, (Lys3-Pro)29, (Orn3-Gly)28 was studied by means of CD. The complexes of these polypeptides with DNA were obtained by the methods of jump-dilution of a two-components mixture from 2 M NaCl to 0.05 M NaCl. The extent of DNA covering by the polypeptides was compared using binding isoterms of ethidium on DNA and DNA-polypeptide complex. The length, L, which polypeptides cover on DNA was estimated by means of energy transfer between the dyes absorbed on the complexes. The CD spectra of the complexes revealed a high sensitivity to changes of the environmental conditions. Small variations in the temperature and ionic strength produces marked changes in the CD spectra of the complexes. It was suggested that observed CD changes are due to both the structural relaxation of the complexes and the existence of liquid-crystal domains in solution.

Chemical Phenomena

[Circular dichroism of DNA complexes with dyes. III. Effect of latent optical activity and the structure of the complexes].

Circular dichroism anisotropy was studied both theoretically and experimentally for the complexes of DNA and dsRNA with dyes (proflavine, 2,7-di-t-butyl proflavine, "Hoechst-33258") and antibiotics (distamycin A, netropsin and olivomycin). Theoretical analysis showed that general features of CD anisotropy, revealed in the previous studies (CD components--delta epsilon parallel to and delta epsilon perpendicular--are ten times or more bigger than the CD-effect without orientation, and delta epsilon parallel to approximately 2 delta epsilon perpendicular) are due to the existence of a specific effect named "latent" optical activity (LOA). This effect can be observed in many cases of non-chiral symmetrical chromophores if they are oriented. The effect is due to the excitation of an electrical dipole transition and a perpendicular magnetic dipole transition (or quadrupole transition) of a molecule. The amplitude and the sign of the LOA-effect depends on the orientation of the chromophores with respect to the light beam; with a random orientation the mutual compensation of LOA-effects of different chromophores happens and no LOA-effect appears. The analitycal expressions relating the value of LOA-effect of the system with electronical characteristics of the chromophores and the geometrical parametra of their arrangement was obtained. The experimental data obtained for the oriented complexes of DNA and dsRNA with proflavine made it possible to determine an angle between the chromophore and the plane perpendicular to the DNA axis--gamma. For the calf thymus DNA gamma = = + 1.8 +/- 0.4 degrees, for the phage T2 DNA gamma = + 2.2 +/- 0.4 degrees, and for phage f2 dsRNA gamma=--3.5 +/- +/- 0.5 degrees. These results, obtained at relatively low concentrations of the bound proflavine (r approximately 0.01), are in accordance with the intercalating mode of the dye binding. A study of CD anisotropy of DNA complexes with other ligands showed that many different chromophores possess LOA-effect. This phenomenon can be used to obtain both spectroscopic and structural information about the systems similar to those reported here.

Acridines