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Balabushevich N.G.,Moscow State University | Pechenkin M.A.,Moscow State University | Lopes de Gerenyu A.V.,Moscow State University | Zorov I.N.,Moscow State University | And 2 more authors.
Moscow University Chemistry Bulletin | Year: 2014

This study was aimed at examination of microparticles formed via the layer-by-layer adsorption of dextran sulfate and chitosan onto the insoluble complexes of various proteins with polyanions. Microparticles with all tested proteins were stable at pH values of 1-5. At pH > 6 the mucoadhesivity of the microparticles changed and the encapsulated proteins were released. Microparticles were able to protect the proteins from proteases. Proteinous protease inhibitors encapsulated as well (2-3%) completely prevented protein proteolysis. The pharmacological effect of microencapsulated insulin was studied in vivo using the model of chronic diabetes in rats, which were treated by oral administration. © 2014 Allerton Press, Inc.

Makarova S.S.,Moscow State University | Minina E.A.,Swedish University of Agricultural Sciences | Makarov V.V.,Moscow State University | Semenyuk P.I.,Moscow State University | And 6 more authors.
Biochimie | Year: 2011

Arabidopsis thaliana At-4/1 is the protein of unknown function capable of polar localization in plant cells and intercellular trafficking. In this work, we cloned cDNAs and chromosomal genes of At-4/1 orthologues from several Nicotiana species. Similarly to the 4/1 genes of A. thaliana and Oryza sativa, Nicotiana 4/1 genes have eight exons and seven introns but are considerably longer due to their larger introns. The allotetraploid genome of Nicotiana tabacum, which is known to consist of the "S genome" originated from Nicotiana sylvestris and the "T genome" derived from Nicotiana tomentosiformis, encodes two 4/1 genes. The T genome-encoded 4/1 gene, but not that of the S genome, contains a SINE-like transposable element in its intron 2. The 4/1 genes of Nicotiana hesperis and Nicotiana benthamiana lack such an element in the intron 2, but possess a related SINE-like sequence in their intron 4. Collectively, the sequence analysis data provide an insight into the organization of 4/1 genes in flowering plants and the patterns of evolution in the genus Nicotiana. The Nicotiana 4/1 proteins and those of other flowering plants show a significant level of sequence similarity. Computer-assisted analysis was further used to compare their predicted secondary structures. Several algorithms confidently predicted the presence of several coiled-coil domains occupying similar positions in different 4/1 proteins. Analysis of circular dichroism spectra carried out for bacterially expressed N. tabacum 4/1 protein (Nt-4/1) and its N- and C-terminally truncated mutants confirmed that the secondary structure of Nt-4/1 is generally alpha-helical. The C-terminal region of Nt-4/1 was found to undergo a partial proteolysis in Escherichia coli cells. Differential scanning calorimetry of Nt-4/1 protein and its mutants revealed three calorimetric domains most probably corresponding to the N-terminal, central, and C-terminal structural domains of the protein. © 2011 Elsevier Masson SAS. All rights reserved.

Balabushevich N.G.,Moscow State University | Vikhoreva G.A.,Moscow State Textile University | Mikhal'chik E.V.,Scientific Research Institute of Physico Chemical Medicine | Larionova N.I.,Moscow State University
Moscow University Chemistry Bulletin | Year: 2010

Thorough investigation and comparative study were conducted for insulin-loaded microparticles fabricated by consecutive adsorption of polyanions (dextran sulfate and chitosan sulfate) and polycations (chitosan and protamine) onto protein microaggregates. The possible regulation of insulin release from the particles by variation in polyelectrolyte pairs, in the number of their adsorption cycles and in pH of media was demonstrated. For all studied cases the microparticles showed protective action towards insulin inactivation at acid pH values and protein release at pH > 5, corresponding to human gastro-intestinal conditions. © 2010 Allerton Press, Inc.

Podgorny O.V.,Russian Academy of Sciences | Polina N.F.,Scientific Research Institute of Physico Chemical Medicine | Babenko V.V.,Scientific Research Institute of Physico Chemical Medicine | Karpova I.Y.,Scientific Research Institute of Physico Chemical Medicine | And 3 more authors.
Journal of Microbiological Methods | Year: 2015

Chlamydia are obligate intracellular parasites of humans and animals that cause a wide range of acute and chronic infections. To elucidate the genetic basis of chlamydial parasitism, several approaches for making genetic modifications to Chlamydia have recently been reported. However, the lack of the available methods for the fast and effective selection of genetically modified bacteria restricts the application of genetic tools. We suggest the use of laser microdissection to isolate of single live Chlamydia-infected cells for the re-cultivation and whole-genome sequencing of single inclusion-derived Chlamydia. To visualise individual infected cells, we made use of the vital labelling of inclusions with the fluorescent Golgi-specific dye BODIPY® FL C5-ceramide. We demonstrated that single Chlamydia-infected cells isolated by laser microdissection and placed onto a host cell monolayer resulted in new cycles of infection. We also demonstrated the successful use of whole-genome sequencing to study the genomic variability of Chlamydia derived from a single inclusion. Our work provides the first evidence of the successful use of laser microdissection for the isolation of single live Chlamydia-infected cells, thus demonstrating that this method can help overcome the barriers to the fast and effective selection of Chlamydia. © 2014 Elsevier B.V.

Dubinkina V.B.,Moscow Institute of Physics and Technology | Tyakht A.V.,Moscow Institute of Physics and Technology | Ilina E.N.,Scientific Research Institute of Physico Chemical Medicine | Ischenko D.S.,Moscow Institute of Physics and Technology | And 12 more authors.
Biomeditsinskaya Khimiya | Year: 2015

Here we present the first metagenomic study of gut microbiota in patients with alcohol dependence syndrome (ADS) performed in the whole-genome ("shotgun") format. Taxonomic analysis highlighted changes in community "drivers" abundance previously associated with inflammatory processes (including increase in Ruminococcus gnavus and torques, as well as decrease in Faecalibacterium and Akkermansia). Microbiota of alcoholics manifested presence of specific opportunistic pathogens rarely detected in healthy control subjects of the world. Differential analysis of metabolic potential basing on changes in KEGG Orthology groups abundance revealed increase in pathways associated with response to oxidative stress. Analysis of two specific gene groups - alcohol metabolism and virulence factors - also showed increase in comparison with the control groups. We suggest that gut microbiota distinct in alcoholics by both taxonomic and functional composition plays role in modulating the effect of alcohol on host organism.

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