Vol 4, No 4 (2012)

Short communications

Letter from Editors

Acta Naturae -.

Abstract

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Acta Naturae. 2012;4(4):1-1
pages 1-1 views

Forum

A Long Road to the Spatial Structure and Mechanisms of Action

Tsetlin V.I.

Abstract

The 2012 Nobel Prize in chemistry has been awarded to Robert Lefkowitz, professor at Duke University Medical Center, and Brian Kobilka, professor at Stanford University School of Medicine, for “research into G protein-coupled receptors.” The wording “research,” rather than “discovery, elucidation, or identification of something,” speaks to the fact that such studies have been under way for nearly 40 years, from almost the zero level to the peaks conquered over the past few years.

Acta Naturae. 2012;4(4):6-7
pages 6-7 views

The 2012 Nobel Prize in Physiology or Medicine

Tomilin A.N.

Abstract

The 2012 Nobel Prize in physiology or medicine has been awarded to John Bertrand Gurdon and Shinya Yamanaka “for the discovery that mature cells can be reprogrammed to become pluripotent.”

Acta Naturae. 2012;4(4):8-10
pages 8-10 views

Made in Russia

Acta Naturae -.

Abstract

By the end of 2012, the Government of the Russian Federation is to approve the State Program “Development of the Pharmaceutical and Medical Industries” for 2013–2020, which includes the current Federal Target-Oriented Program “Pharma-2020.” One of the objectives within the State Program prepared by the Ministry of Industry and Trade is to “increase the share of domestically produced drugs and medicinal products in overall consumption by the public healthcare services of the Russian Federation by 48%.” However, the term “domestically produced drug” still remains to be legislatively defined. According to the draft resolution issued by the Ministry of Industry and Trade in May 2012, a “domestic drug” should mean a drug whose production cycle in the territory of the Russian Federation starts from a substance or a ready-toconsume formulation. Until 2014, the Ministry was ready to regard even those drugs whose packaging was made in Russia as Russian ones. However, no further steps followed. Therefore, the question pertaining to which drugs and which produced by which pharmaceutical companies should be regarded as domestic drugs remains open. Actors of the Russian pharmaceutical industry share their opinions.

Acta Naturae. 2012;4(4):11-16
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Reviews

Recombinant Influenza Vaccines

Sedova E.S., Shcherbinin D.N., Migunov A.I., Smirnov Y.A., Logunov D.Y., Shmarov M.M., Tsybalova L.M., Naroditskiĭ B.S., Kiselev O.I., Gintsburg A.L.

Abstract

This review covers the problems encountered in the construction and production of new recombinant influenza vaccines. New approaches to the development of influenza vaccines are investigated; they include reverse genetics methods, production of virus-like particles, and DNA- and viral vector-based vaccines. Such approaches as the delivery of foreign genes by DNA- and viral vector-based vaccines can preserve the native structure of antigens. Adenoviral vectors are a promising gene-delivery platform for a variety of genetic vaccines. Adenoviruses can efficiently penetrate the human organism through mucosal epithelium, thus providing long-term antigen persistence and induction of the innate immune response. This review provides an overview of the practicability of the production of new recombinant influenza cross-protective vaccines on the basis of adenoviral vectors expressing hemagglutinin genes of different influenza strains.

Acta Naturae. 2012;4(4):17-27
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Epigenetics of Pluripotent Cells

Medvedev S.P., Pokushalov E.A., Zakian S.M.

Abstract

Pluripotency is maintained by a complex system that includes the genetic and epigenetic levels. Recent studies have shown that the genetic level (transcription factors, signal pathways, and microRNAs) closely interacts with the enzymes and other specific proteins that participate in the formation of the chromatin structure. The interaction between the two systems results in the unique chromatin state observed in pluripotent cells. In this review, the epigenetic features of embryonic stem cells and induced pluripotent stem cells are considered. Special attention is paid to the interplay of the transcription factors OCT4, SOX2, and NANOG with the Polycomb group proteins and other molecules involved in the regulation of the chromatin structure. The participation of the transcription factors of the pluripotency system in the inactivation of the X chromosome is discussed. In addition, the epigenetic events taking place during reprogramming of somatic cells to the pluripotent state and the problem of “epigenetic memory” are considered.

Acta Naturae. 2012;4(4):28-46
pages 28-46 views

Research Articles

Effect of 3D Cultivation Conditions on the Differentiation of Endodermal Cells

Petrakova O.S., Ashapkin V.V., Voroteliak E.A., Bragin E.Y., Shtratnikova V.Y., Chernioglo E.S., Sukhanov Y.V., Terskikh V.V., Vasiliev A.V.

Abstract

Cellular therapy of endodermal organs is one of the most important issues in modern cellular biology and biotechnology. One of the most promising directions in this field is the study of the transdifferentiation abilities of cells within the same germ layer. A method for an in vitro investigation of the cell differentiation potential (the cell culture in a three-dimensional matrix) is described in this article. Cell cultures of postnatal salivary gland cells and postnatal liver progenitor cells were obtained; their comparative analysis under 2D and 3D cultivation conditions was carried out. Both cell types have high proliferative abilities and can be cultivated for more than 20 passages. Under 2D cultivation conditions, the cells remain in an undifferentiated state. Under 3D conditions, they undergo differentiation, which was confirmed by a lower cell proliferation and by an increase in the differentiation marker expression. Salivary gland cells can undergo hepatic and pancreatic differentiation under 3D cultivation conditions. Liver progenitor cells also acquire a pancreatic differentiation capability under conditions of 3D cultivation. Thus, postnatal salivary gland cells exhibit a considerable differentiation potential within the endodermal germ layer and can be used as a promising source of endodermal cells for the cellular therapy of liver pathologies. Cultivation of cells under 3D conditions is a useful model for the in vitro analysis of the cell differentiation potential.

Acta Naturae. 2012;4(4):47-57
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N-Terminal Fusion Tags for Effective Production of G-Protein-Coupled Receptors in Bacterial Cell-Free Systems

Lyukmanova E.N., Shenkarev Z.O., Khabibullina N.F., Kulbatskiy D.S., Shulepko M.A., Petrovskaya L.E., Arseniev A.S., Dolgikh D.A., Kirpichnikov M.P.

Abstract

G-protein-coupled receptors (GPCR) constitute one of the biggest families of membrane proteins. In spite of the fact that they are highly relevant to pharmacy, they have remained poorly explored. One of the main bottlenecks encountered in structural-functional studies of GPCRs is the difficulty to produce sufficient amounts of the proteins. Cell-free systems based on bacterial extracts from E. coli cells attract much attention as an effective tool for recombinant production of membrane proteins. GPCR production in bacterial cell-free expression systems is often inefficient because of the problems associated with the low efficiency of the translation initiation process. This problem could be resolved if GPCRs were expressed in the form of hybrid proteins with N-terminal polypeptide fusion tags. In the present work, three new N-terminal fusion tags are proposed for cellfree production of the human β2-adrenergic receptor, human M1 muscarinic acetylcholine receptor, and human somatostatin receptor type 5. It is demonstrated that the application of an N-terminal fragment (6 a.a.) of bacteriorhodopsin from Exiguobacterium sibiricum (ESR-tag), N-terminal fragment (16 а.о.) of RNAse A (S-tag), and Mistic protein from B. subtilis allows to increase the CF synthesis of the target GPCRs by 5–38 times, resulting in yields of 0.6–3.8 mg from 1 ml of the reaction mixture, which is sufficient for structural-functional studies.

Acta Naturae. 2012;4(4):58-64
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Identification of Novel RNA-Protein Contact in Complex of Ribosomal Protein S7 and 3’-Terminal Fragment of 16S rRNA in E. coli

Golovin A.V., Khayrullina G.A., Kraal B., Kopylov А.М.

Abstract

For prokaryotes in vitro, 16S rRNA and 20 ribosomal proteins are capable of hierarchical self- assembly yielding a 30S ribosomal subunit. The self-assembly is initiated by interactions between 16S rRNA and three key ribosomal proteins: S4, S8, and S7. These proteins also have a regulatory function in the translation of their polycistronic operons recognizing a specific region of mRNA. Therefore, studying the RNA–protein interactions within binary complexes is obligatory for understanding ribosome biogenesis. The non-conventional RNA–protein contact within the binary complex of recombinant ribosomal protein S7 and its 16S rRNA binding site (236 nucleotides) was identified. UV–induced RNA–protein cross-links revealed that S7 cross-links to nucleotide U1321 of 16S rRNA. The careful consideration of the published RNA– protein cross-links for protein S7 within the 30S subunit and their correlation with the X-ray data for the 30S subunit have been performed. The RNA – protein cross–link within the binary complex identified in this study is not the same as the previously found cross-links for a subunit both in a solution, and in acrystal. The structure of the binary RNA–protein complex formed at the initial steps of self-assembly of the small subunit appears to be rearranged during the formation of the final structure of the subunit.

Acta Naturae. 2012;4(4):65-72
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Carbocyclic Analogues of Inosine-5’-Monophosphate: Synthesis and Biological Activity

Matyugina E.S., Andreevskaya S.N., Smirnova T.G., Khandazhinskaya A.L.

Abstract

9-(4’-Phosphonomethoxy-2’-cyclopenten-1’-yl)hypoxanthine and 9-(4’-phosphonomethoxy-2’,3’-dihydroxycyclopenten-1’-yl)hypoxanthine were synthesized as isosteric carbocyclic analogues of inosine-5’-monophosphate. The synthesized compounds were shown to be capable of inhibiting the activity of human type II inosine-5′-monophosphate dehydrogenase (IMPDH II) (IC50 = 500 μM) and to have no significant effects on the growth of Mycobacterium tuberculosis.

Acta Naturae. 2012;4(4):73-77
pages 73-77 views

Gold Nanoparticle Clusters in Quasinematic Layers of Liquid-Crystalline Dispersion Particles of Double-Stranded Nucleic Acids

Yevdokimov Y.M., Salyanov V.I., Katz E.I., Skuridin S.G.

Abstract

This work is devoted to the investigation of the methanogenic archaea involved in anaerobic digestion of cattle manure and maize straw on the basis of terminal restriction fragment length polymorphism (TRFLP) analysis of archaeal 16S rRNA genes. The biological diversity and dynamics of methanogenic communities leading to anaerobic degradation of agricultural organic wastes with biogas production were evaluated in laboratory-scale digesters. T-RFLP analysis, along with the establishment of archaeal 16S rRNA gene clone libraries, showed that the methanogenic consortium consisted mainly of members of the genera Methanosarcina and Methanoculleus, with a predominance of Methanosarcina spp. throughout the experiment.

Acta Naturae. 2012;4(4):78-90
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Methanogenic Community Dynamics during Anaerobic Utilization of Agricultural Wastes

Ziganshin A.M., Ziganshina E.E., Kleinsteuber S., Pröter J., Ilinskaya O.N.

Abstract

This work is devoted to the investigation of the methanogenic archaea involved in anaerobic digestion of cattle manure and maize straw on the basis of terminal restriction fragment length polymorphism (TRFLP) analysis of archaeal 16S rRNA genes. The biological diversity and dynamics of methanogenic communities leading to anaerobic degradation of agricultural organic wastes with biogas production were evaluated in laboratory-scale digesters. T-RFLP analysis, along with the establishment of archaeal 16S rRNA gene clone libraries, showed that the methanogenic consortium consisted mainly of members of the genera Methanosarcina and Methanoculleus, with a predominance of Methanosarcina spp. throughout the experiment.

Acta Naturae. 2012;4(4):91-97
pages 91-97 views

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