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[Comparative study of the toxicity of poly G-poly C and poly I-poly C in different objects].

The poly(G).poly(C) complex has the same interferon-inducing and antiviral activity upon parenteral administration to white mice as poly(I).poly(C), but is considerably less toxic. Upon intravenous inoculation of poly(I).poly(C) to mice its LD50 is 15.8 mg/kg whereas poly(G).poly(C) is not toxic in doses up to 200 mg/kg. In rabbits inoculated with poly(I).ploy(C) intravenously its LD50 is 0.22 mg/kg, while poly(G).poly(C) is not toxic in doses of 1 mg/kg. Histological examinations of different organs of mice and rats revealed no pathomorphological changes after a single intravenous and intraperitoneal inoculation of poly(G).poly(C). It exerted no embryotoxic effect in mice in a dose of 5 mg/kg and was considerably less toxic than poly(I).poly(C) in continuous diploid cell cultures of human embryo lung cells.

Animals↗

[Effect of the size of the continuous poly(G) site in poly (G, A).poly (C) complexes on their interferon-inducing activity and their capacity to stimulate the development of immunity].

It was established that the level of interferon-inducing activity of poly(G90A1).poly(C) complex in cell cultures and in mice was comparable to that of poly(G).poly(C). As the size of the continuous sites of poly(G) in the purine strand in poly(G, A).poly(C) complexes decreased to 60 and 28 nucleotides, the interferon-inducing activity decreased progressively, was still marked, or approached the zero at G:A ratios equal to 17:1 or 10:1, respectively. All this indicates that the cell receptors responsible for switching on of the induction mechanisms for interferon synthesis still recognize the stimulus 17 and possibly less so stimulus 10 of successively located guanosine nucleotides complementary to poly(C) and provide for the highest interferon production level when their number is equal to or exceeds 90-100. With the exception of poly(G10A1).poly(C) in which the interferon-inducing activity did not exceed its detection threshold, all complexes enhanced noticeably or markedly specific immune response in mice to tick-borne encephalitis after immunization with inactivated unadsorbed tissue culture vaccine against this infection. The level of this immunostimulating activity correlated irregularly with the intensity of their interferon-inducing activity.

Adjuvants, Immunologic↗

[Laboratory and clinical study of biological activity of poly G-poly C complex].

Poly(G).poly(C) inoculated intravenously to mice in a dose of 100 microgram induced interferon in the blood in amounts comparable to those induced by poly(I).poly (C). In contrast to rapid accumulation (within 2 hours after induction) and rapid disappearance of interferon in response to poly(I).poly(C) inoculation, the interferon induced by poly(G).poly(C) reached the maximum titer by 6 hours and remained at a high level for 24 hours after inoculation. When given to human volunteers intranasally in a dose of 6 mg, the poly(G).poly(C) complex induced interferon in the blood serum in 70% of the subjects in a titer of 85 units/ml within 24 hours.

Administration, Intranasal↗

[Modification of the poly G-poly C complex by incorporation of adenosine in the purine chain].

Antiviral and interferonogenic activity of the complexes of poly(G,A) . poly(C) and poly(G) . poly(C) was studied in mice and cell cultures. Three out of 4 complexes of poly(G,A) . poly(C) had insignificant antiviral and interferonogenic activity in chick embryo cells. One of the complexes induced low levels of interferon production in mice and decreased the rate of their death from experimental forest-spring encephalitis. The activity of poly(G) . poly(C) in the above cell systems was much more pronounced. Unlike this complex, some complexes of poly(G,A) . poly(C) showed a noticeable activity in the cells of Primates. The effect of the noncomplementary base in the purine thread of poly(G) . poly(C) on its biological activity and nucleotide composition is discussed.

Adenosine↗

Template-dependent biosynthesis of poly(G) x poly (C) and its antiviral activity in vitro and in vivo.

Experimental conditions for poly(G) synthesis from GTP on a poly(C) template with the aid of Escherichia coli DNA-dependent RNA polymerase were investigated. The reaction product was purified without the use of RNase. On the basis of spectral data, gel permeation chromatography, affinity adsorption and electron microscopic visualization, the poly(G) x poly(C) product was assumed to possess a high degree of structural regularity. Its in vitro and in vivo antiviral activities were compared with those of traditional poly(G) x poly(C) and poly(I) x poly(C). Template-dependent poly(G) x poly(C) was similar in its in vitro activity to poly(I) x poly(C) or even surpassed it, whereas the 'traditional' poly(G) x poly(C) was only slightly active in vitro. However, 'traditional' poly(G) x poly(C) and poly(I) x poly(C) had similar activity in vivo, whereas template-dependent poly(G) x poly(C) was much less active in vivo. The role of intramolecular structural regularity in the in vitro and in vivo antiviral activity of polyribonucleotide duplexes is discussed.

Animals↗

[Effect of virazole on the antiviral activity of poly(G) X poly(C) and other polyribonucleotide interferonogens].

The effect of virazole on the antiviral activity of poly (G) X poly (C), poly (G, A) X X poly (C) and poly(G, I) X poly (C) was studied in cell cultures and on mice. It was shown that virazole in concentrations not sufficient for significant inhibition of the development of vesicular stomatitis virus or Sindbis virus in chick embryo cell cultures markedly increased the antiviral effect and allowed decreasing the minimum effective doses of the synthetic polyribonucleotide complexes with respect to the above viruses. Combined administration of poly (G) X poly (C) and virazole to mice 1-2 or 24 hours after infection with tick-borne encephalitis virus provided a much more pronounced decrease in the death rate of the animals than the use of the interferonogen alone. Virazole per se was little active and had no significant effect on the intensity of interferonogenesis promoted by the use of poly (G) X poly (C). A possibility of successful therapy of viral infections with polyribonucleotide interferonogens in combination with virazole or other chemotherapeutic drugs with broad antiviral spectrum is discussed.

Animals↗

[Evaluation of the size of the continuous poly(G) site necessary for the biological activity of the poly(G).poly(C) complex].

On the basis of synthesis of a series of poly(G, A).poly(C) copolymers with changing G:A ratio from 15:1 to 90:1 and trials of their biological activity in comparison with poly(G).poly(C), the size of poly(G) in it was evaluated within the range of a continuous double-stranded area necessary for the activity. The antiviral activity close to that of poly(G).poly(C) in experimental tick-borne encephalitis of mice and vesicular stomatitis virus infection of chick embryo cells was found only in poly(G,A).poly(C) complexes with a G:A ratio equal to or higher than 90:1. Consequently, the high activity of poly(G).poly(C) is present at an average length of poly(G) equal to 90-100 nucleotides within the limits of the continuous double-stranded area.

Animals↗

Preparation and biological properties of a highly active poly(G) X poly(C) inducer of interferon.

Experimental conditions necessary for the full expression of interferon-inducing activity by a complex of polyguanylic acid and polycytidylic acid included: (i) a sufficiently high molecular size of each homopolymer; (ii) annealing conditions which insured complete denaturation of polyguanylic acid self-structure; and (iii) the specific biological assay employed to assay interferon-inducing potency. The complex of polyguanylic acid and polycytidylic acid possessed several properties that suggested it may be an atypical polynucleotide interferon inducer. For instance, it was inactive in primary rabbit kidney cell cultures, usually exquisitely sensitive to polynucleotide interferon inducers, unless it was incubated on the cell cultures for prolonged times or in the presence of DEAE-dextran. Polyguanylic acid X polycytidylic acid could induce interferon in rabbits and mice but gave a more protracted response than did poly(I) X poly(C). Finally, poly(G) X poly(C) was, without any modification, resistant to degradation by serum nucleases.

Animals↗

[Nuclear proteins from Drosophila melanogaster specifically binding to simple homopolymeric sequences of poly[d(T-G)].poly[d(C-A)] type].

Binding of simple homopolymeric sequences to Drosophila melanogaster nuclear proteins has been studied. Proteins with Mr 65-72 kDa have been found, which specifically bind to synthetic poly[d(T-G)].poly[d(C-A)], as well as to D. melanogaster DNA containing a block of poly[d(T-G)].poly[d(C-A) 40 b.p. in length. It has been shown, that these proteins bind only to poly[d(T-G).poly[d(C-A)] and not to other types of simple sequences, for example poly[d(G-A)].poly[d(T-C)] and poly[d(A-T)].

Animals↗

Pancreatic ribonuclease-poly G complexes: complete inhibition of poly U hydrolysis.

Pancreatic ribonuclease forms large complexes with poly G in 0.1 M acetate buffer solutions (pH 5.4). These are largest when the ratio, of ribonuclease to poly G concentration, is slightly less than 2. Under the same conditions lysozyme forms still larger complexes with poly U, and these are largest when the ratio, of lysozyme to poly U concentration, is about 2.5. The ribonuclease in ribonuclease-poly G complexes digests poly U. Free ribonuclease digests the poly U in lysozyme-poly U complexes. However, when the poly G concentration is about an order of magnitude greater than that required to bind all the ribonuclease, lysozyme-bound poly U is not hydrolyzed.

Acetates↗

Interaction of poly(L-lysine)-g-poly(ethylene glycol) with supported phospholipid bilayers.

Interactions between the graft copolymer poly(L-lysine)-g-poly(ethylene glycol), PLL-g-PEG, and two kinds of surface-supported lipidic systems (supported phospholipid bilayers and supported vesicular layers) were investigated by a combination of microscopic and spectroscopic techniques. It was found that the application of the copolymer to zwitterionic or negatively charged supported bilayers in a buffer of low ionic strength led to their decomposition, with the resulting formation of free copolymer-lipid complexes. The same copolymer had no destructive effect on a supported vesicular layer made up of vesicles of identical composition. A comparison between poly(L-lysine), which did not induce decomposition of supported bilayers, and PLL-g-PEG copolymers with various amounts of PEG side chains per backbone lysine unit, suggested that steric repulsion between the PEG chains that developed upon adsorption of the polymer to the nearly planar surface of a supported phospholipid bilayer (SPB) was one of the factors responsible for the destruction of the SPBs by the copolymer. Other factors included the ionic strength of the buffer used and the quality of the bilayers, pointing toward the important role defects present in the SPBs play in the decomposition process.

Adsorption↗

Poly(N-isopropylacrylamide)-g-poly(ethyleneoxide) for high resolution and high speed separation of DNA by capillary electrophoresis.

A new separation medium, poly(N-isopropylacrylamide)-g-poly(ethyleneoxide) (PNI-PAM-g-PEO) solution, used for double-stranded (ds) DNA separation by capillary electrophoresis (CE) is presented. This type of grafted copolymer has a good self-coating ability for quartz capillary tubing and a slightly temperature-dependent viscosity-adjustable property, making it easier to use. One bp resolution was achieved within 12.5 min by using 8% w/v PNIPAM-gPEO in 1 x TBE (Tris-borate-ethylenediaminetetraaceticacid) buffer with an effective column length of 10 cm and an applied electric field strength of 200 V/cm. The PNIPAM-g-PEO solutions had a high sieving ability for relatively small sized DNAs with the relative standard derivation for the first 10 runs being less than 0.9% by using the same polymer solution. With 8% w/v PNIPAM-g-PEO solution in a 1.5 cm column and 2400 V as the running voltage, phiX174/HaeIII digest could be clearly separated within 24 s.

Acrylamides↗