In Search of New Inhibitors of HIV-1 Replication: Synthesis and Study of 1-(2′-Deoxy-β-D-Ribofuranosyl)-1,2,4-Triazole-3-Carboxamide as a Selective Viral Mutagenic Agent

With the emergence of HIV strains resistant or cross-resistant to nearly all antiretroviral regimen, novel therapy approaches have to be considered. As a part of our current work on viral mutagenic compounds, we prepared 1-(2′ -deoxy-β-D-ribofuranosyl)-1,2,4-triazole-3-carboxamide (2′ -deoxy-ribavirin) and its 5′ -triphosphate derivative. The nucleoside mutagenic activity was evaluated on HIV-1 NL4-3 in CEMx174 cell culture. After 2.5 months, no reduction on HIV-1 viability was observed. On the other hand, in vitro experiments with purified HIV-1 RT demonstrated that the triphosphate analog can be incorporated opposite to several natural nucleosides.     

Investigation of the Synthesis of 2-Carbamoylmethyl-6-D-Ribofuranosylpyridine

Abstract

During the course of our study on the synthesis of new pyridine-C-nucleosides with potential anti-viral and/or anti-tumourai properties, the preparation of 2-carbamoylmethyl-6-β-D-ribofuranosylpyridine 1, by means of a lithiation reaction on 2-(2,3-0-isopropylidene-β D-nbofuranosyl)-6-methylpyridine 2, was investigated (Scheme I). The latter compound was prepared by treating 6-methyl- toluenesulphonic acid. 2-(& D-ribofuranosyl)pyridine 1 2 with 2.2-dimethoxypropane in the presence of p-     

Arenesulfonylethoxycarbonyl- a Set of Amino Protecting Groups for DNA and RNA Synthesis

Abstract

Phenyl-, 4-chlorophenyl- and 4-nitrophenylsufonylethoxycarbonyl groups have been reported for the first time as the exocyclic amino protecting groups in nucleoside chemistry. They are all stable under the standard conditions of manipulations in phosphotriester and phosphiteamidite chemistry, they are removable both under the alkaline hydrolytic conditions and also under the influence of non-nucleophilic tertiary bases. N³-Phenyl- and 4-toluenesulfonylethoxycarbonyl derivatives of uridine have been also prepared and characterized by ¹⁵N-NMR spectroscopy, their stabilities under different conditions have been tested.     

Achiral Nucleotide Analogs: 5,5-Disubstituted Pyrimidines Related to Barbituric Acid

Abstract

Achiral nucleotide analogs are of potential interest in origin of life studies. Four new 5,5-diphosphoethylpyrimidine analogs were prepared by condensation of urea or guanidine with a, a-di(2-[ethoxy-I-ethoxy]ethyl)malononitrile or a, a-di(2-[ethoxy-1-ethoxy]ethyl)ethyl cyanoacetate respectively, followed by removal of the protecting groups and bisphosphorylation. Oligomers of two potentially complementary analogs have been prepared. Interactions between these oligomers appear to be weak in aqueous solution.     

Relative Effects of Graft Copolymer and Polyamines on Triplex Stabilization Under Physiological Conditions

Abstract

Triplex-stabilizing effect of a graft copolymer under physiologically relevant conditions has been evaluated and compared with other polyamines. Here we show that the graft copolymer significantly stabilizes triplex DNAs with amazingly higher efficicacy than that of physiological concentrations of spermine and spermidine.     

Expression of cecropin P1 gene increases resistance of Camelina sativa (L.) plants to microbial phytopathogenes

Transgenic plants of camelina (Camelina sativa (L.) Crantz) with the synthetic gene of antimicrobial peptide cecropin P1 (cecP1) were obtained. Agrobacterium-mediated transformation is performed using the binary vector pGA482::cecP1 by vacuum infiltration of flower buds. The presence of the cecP1 gene in the genome of plants was confirmed by PCR. CecP1 gene expression in transgenic plants was shown by Western blot analysis and by antimicrobial activity of plant extracts against the bacterial phytopathogene Erwinia carotovora. The plants of F₀ and F₁ generations had the normal phenotype and retained the ability to form viable seeds in self-pollination. cecP1 plants exhibit enhanced resistance to bacterial and fungal phytopathogens: Erwinia carotovora and Fusarium sporotrichioides. The increased sustainability of cecropin P1-expressing plants against salt stress is shown. The possibility of the integration of the cecP1 gene into the overall protective system of plants against biotic and abiotic stresses is discussed.     

Genetic diversity of Besermyans inferred from mitochondrial DNA polymorphism

The first data are presented on mtDNA diversity in Besermyans, the Finno-Ugric ethnic group related to Udmurts. An analysis of mtDNA polymorphism showed that Besermyans stood out from the other populations of Volga-Ural region due to the presence of a large proportion of the Mongoloid component. The sample of Besermyans contained East Eurasian haplotypes not detected in ethnic populations of the Volga region and Cisurals, while they were detected in South Siberia, mostly among Turkic-speaking populations. An analysis of the genetic distances between Besermyans and the neighboring ethnic groups showed that Besermyans were distant from other populations of Volga-Ural region and close to Turkic-speaking populations of South Siberia. Thus, the data obtained favor the suggestion on the mixed Udmurto-Turkic origin of Besermyans.     

Effect of disruption of Pichia pastoris YPS1 gene on viability and production of recombinant proteins

In the present study, we constructed a Pichia pastoris mutant strain PS111 that lacks one member of the yapsin family through disruption of the YPS1 gene coding for aspartic protease yapsin 1. Under normal growth conditions, the PS111 mutant strain did not show detectable growth defects. Unlike the S. cerevisiae yps1 mutant, the P. pastoris PS111 strain showed no sensitivity when grown in the presence of CaCl₂, elevated temperature (37°C), under acid (pH 4.9) and alkaline (pH 8.3) conditions. Unlike the S. cerevisiae, the P. pastoris yps1 mutant showed decreased growth phenotype induced by cell wall-perturbing reagent sodium dodecyl sulfate (SDS) only when the concentration of SDS was increased by ten times. The use of the yps1 disruptant to produce human interferon alpha16 (hINF-α16) prevents proteolysis, which occurs in the wildtype strain. It was found that the degradation of recombinant protein Alburon composed of human serum albumin (HSA) and hINF-α16 was slightly decreased in the strain lacking yapsin 1.