Screening studies of POP levels in fish from selected lakes in the Paz watercourse

Appendix 8/15 of the publication "State of the environment in the Norwegian, Finnish and Russian border area 2007" (The Finnish Environment 6/2007)

Противокоронавирусные свойства брассиностероидов

Antiviral properties of natural brassinosteroids of the campestane, ergostane, and stigmastane series (6-ketones and B-lactones) and their (22S,23S)-analogs were studied using the seasonal human respiratory alpha-coronavirus 229E (HCoV-229E) as an example. The presence of anticoronavirus properties was shown for a number of studied compounds. In general, 6-ketones were more active than B-lactones. The maximum inhibitory effect (EC50 21.1 µM) in relation to the reproduction of HCoV-229E was noted for (22S,23S)-epicastasterone.На примере сезонного респираторного альфа-коронавируса человека 229Е (HCoV-229E) проведено изучение противовирусных свойств природных брассиностероидов кампестанового, эргостанового и стигмастанового рядов (6-кетонов и B-лактонов) и их соответствующих (22S,23S)-аналогов. Показано наличие противокоронавирусных свойств для ряда изученных соединений. 6-Кетоны в целом оказались более активными в сравнении с B-лактонами. Максимальный показатель ингибирующего действия (EC50 21,1 µM) в отношении репродукции HCoV-229E отмечен для (22S,23S)-эпикастастерона

Probiotic consortiums: Structure and antagonistic activity against opportunistic bacteria and human normobiota (using the example of <i>Escherichia coli</i>) <i>in vitro</i>

Background. Using probiotic preparations based on consortia of microorganisms not only helps to restore the balance of the intestinal microbiota, but also increases the therapeutic effect of probiotics. Promising sources for obtaining probiotic consortia are milk products that have undergone natural fermentation with the help of spontaneously formed microbial consortia. The aim. To study the structure of five microbial consortia with probiotic properties from naturally fermented milk products and to assess in vitro their antagonistic activity against opportunistic bacteria and a representative of the human normobiota – Escherichia coli. Materials and methods. The structure of bacterial consortia was analyzed by sequencing methods. The antagonistic activity of the consortia was assessed by the disk diffusion method. Results. It has been established that the studied microbial consortiums are represented by Enterococcus spp. and Streptococcus spp. bacteria. In consortiums No. 1, No.  2, and No.  3, Enterococcus bacteria dominated, while in consortiums No.  4 and No. 5, Streptococcus dominated. Antagonistic activity was shown against four isolates of opportunistic bacteria: Klebsiella pneumoniae No.  493, Enterobacter hormaechei No. 372, Staphylococcus aureus No. 4 and Pseudomonas aeruginosa No. 25 IMB, as well as against one representative of the human normobiota – Escherichia coli No. 495. The highest growth delay zone is found in E. coli No. 495 isolate. Three test cultures (K. pneumoniae No. 509, E. coli ATCC25922 and P. aeruginosa No. 3 IMB) exhibited more dense growth around probiotic consortia. Conclusion. The results of the study showed that the effect of probiotic consortia differing in the composition of microorganisms can be neutral and bactericidal. The presence of antagonistic activity in the studied microbial consortia against multiresistant isolates of opportunistic bacteria is a prospect for creating probiotics with antibacterial properties

Ribosomal DNA as DAMPs Signal for MCF7 Cancer Cells

Introduction: The cell free ribosomal DNA (cf-rDNA) is accrued in the total pool of cell free DNA (cfDNA) in some non-cancer diseases and demonstrates DAMPs characteristics. The major research questions: (1) How does cell free rDNA content change in breast cancer; (2) What type of response in the MCF7 breast cancer cells is caused by cf-rDNA; and (3) What type of DNA sensors (TLR9 or AIM2) is stimulated in MCF7 in response to the action of cf-rDNA?Materials and Methods: CfDNA and gDNA were isolated from the blood plasma and the cells derived from 38 breast cancer patients and 20 healthy female controls. The rDNA content in DNA was determined using non-radioactive quantitative hybridization. In order to explore the rDNA influence on MCF7 breast cancer cells, the model constructs (GC-DNAs) were applied: pBR322-rDNA plasmid (rDNA inset 5836 bp long) and pBR322 vector. ROS generation, DNA damage, cell cycle, expression of TLR9, AIM2, NF-kB, STAT3, and RNA for 44 genes affecting the cancer cell viability were evaluated. The methods used: RT-qPCR, fluorescent microscopy, immunoassay, flow cytometry, and siRNA technology.Results: The ratio R = cf-rDNA/g-rDNA for the cases was higher than for the controls (median 3.4 vs. 0.8, p &lt; 10−8). In MCF7, GC-DNAs induce a ROS burst, DNA damage response, and augmentation of NF-kB and STAT3 activity. The number of the apoptotic cells decreases, while the number of cells with an instable genome (G2/M– arrest, micronuclei) increase. Expression of anti-apoptotic genes (BCL2, BCL2A1, BCL2L1, BIRC3, MDM2) is elevated, while expression of pro-apoptotic genes (BAX, BID, BAD, PMAIP1, BBC3) is lowered. The cells response for pBR322-rDNA is much more intense and develops much faster, than response for pBR322, and is realized through activation of TLR9- MyD88 - NF-kB- signaling. This difference in response speed is owing to the heightened oxidability of pBR322-rDNA and better ability to penetrate the cell. Induction of TLR9 expression in MCF7 is followed by blocking AIM2 expression.Conclusion: (1) Ribosomal DNA accumulates in cfDNA of breast cancer patients; (2) Cell free rDNA induce DNA damage response and stimulates cells survival, including cells with an instable genome; (3) Cell free rDNA triggers TLR9- MyD88- NF-kB- signaling, with significantly repressing the expression of AIM2

Электрон-избыточные металлoфенантроцианины – новый класс тетраазахромофорных комплексов d-элементов

Data of experimental and theoretical study (DFT method) of electron-rich metal phenanthrocyanines – new class of tetraazachromophore complexes of delements Pt2+, Pd2+, Rh3+, Cr3+, Co2+, Ni2+, Zn2+ and Cd2+ was presented. On basis of experimental data on electron absorption spectra (EAS) and quantum chemical calculations of EAS of model anion-radical 1,10-phenanthroline and electron-rich 1,10-phenanthroline complexes suggest of long-wave bands in EAS of metal phenanthrocyanines was made. Some general regularities of new metal phenanthrocyanines formation in C(sp2 )H-C(sp2 )H-coupling reactions of coordinated 1,10-phenanthrolines concerned with metal promotion and elementary electron and proton transfer processes in initialization stage of C(sp2 )H-C(sp2 )H-coupling was analysedредставлены результаты экспериментального и теоретического (методом DFT) исследования электрон-избыточных металлофенантроцианинов – нового класса тетраазахромофорных комплексов d-элементов Pt2+, Pd2+, Rh3+, Cr3+, Co2+, Ni2+, Zn2+ и Cd2+. Сделаны отнесения длинноволновых полос в ЭСП металлофенантроцианинов. Проанализированы некоторые общие закономерности образования новых металлофенантроцианинов в реакциях C(sp2 )H-C(sp2 )H-сочетания координированных 1,10-фенантролинов, связанные с металл-промотированием, а также с элементарными процессами электронного и протонного переноса на стадии инициирования C(sp2 )H-C(sp2 )H-сочетания

Dual Mechanism for the Translation of Subgenomic mRNA from Sindbis Virus in Infected and Uninfected Cells

Infection of BHK cells by Sindbis virus (SV) gives rise to a profound inhibition of cellular protein synthesis, whereas translation of viral subgenomic mRNA that encodes viral structural proteins, continues for hours. To gain further knowledge on the mechanism by which this subgenomic mRNA is translated, the requirements for some initiation factors (eIFs) and for the presence of the initiator AUG were examined both in infected and in uninfected cells. To this end, BHK cells were transfected with different SV replicons or with in vitro made SV subgenomic mRNAs after inactivation of some eIFs. Specifically, eIF4G was cleaved by expression of the poliovirus 2A protease (2Apro) and the alpha subunit of eIF2 was inactivated by phosphorylation induced by arsenite treatment. Moreover, cellular location of these and other translation components was analyzed in BHK infected cells by confocal microscopy. Cleavage of eIF4G by poliovirus 2Apro does not hamper translation of subgenomic mRNA in SV infected cells, but bisection of this factor blocks subgenomic mRNA translation in uninfected cells or in cell-free systems. SV infection induces phosphorylation of eIF2α, a process that is increased by arsenite treatment. Under these conditions, translation of subgenomic mRNA occurs to almost the same extent as controls in the infected cells but is drastically inhibited in uninfected cells. Notably, the correct initiation site on the subgenomic mRNA is still partially recognized when the initiation codon AUG is modified to other codons only in infected cells. Finally, immunolocalization of different eIFs reveals that eIF2 α and eIF4G are excluded from the foci, where viral RNA replication occurs, while eIF3, eEF2 and ribosomes concentrate in these regions. These findings support the notion that canonical initiation takes place when the subgenomic mRNA is translated out of the infection context, while initiation can occur without some eIFs and even at non-AUG codons in infected cells

CHARACTERISTICS OF THE BAIKAL SUBTYPE OF TICK-BORNE ENCEPHALITIS VIRUS CIRCULATING IN EASTERN SIBERIA

Background. During the study of the genetic variability of the tick-borne encephalitis virus (TBEV) in Eastern Siberia, a group of 22  strains with a unique genetic structure significantly different from all  known TBEV subtypes was identified. This TBEV variant was  tentatively called “group 886”. Therefore, for this original TBEV  variant it was necessary to study the genetic, biological properties of the “group 886” strains, clarify its TBEV taxonomic status, its range, evolutionary history, etc.Aim. The generalization of the currently available data on genetic and biological properties of TBEV “886” group.Materials and methods. The genetic structure of “group 886” strains was studied by the complex of molecular-genetic methods (MHNA, sequencing of fragments or the complete genome).Results. It was shown that “group 886” strains form a separate cluster on phylogenetic tree, and the level of genetic differences  from other genotypes is more than 12 %. It was defined that this  TBEV variant has its own area (Irkutsk region, Republic of Buryatia,  Trans-Baikal region, Northern Mongolia). Its ecological connection  with all links of the transmissive chain (ixodid ticks, small mammals,  human), participation in human pathology, stability and duration of  circulation in the Baikal region, individual evolutionary history  were proved. Some phenotypic characteristics of the “group 886” strains were considered.Conclusion. The presented data testify to the validity of the “886 group” isolation as an independent genetic type. Taking into account  the geographical distribution of this TBEV genotype, we propose to assign it the name “Baikal genotype/subtype”

Electron-rich metal phenanthrocyanine – new class of tetraazachromophor complexes of d-elements

Data of experimental and theoretical study (DFT method) of electron-rich metal phenanthrocyanines – new class of tetraazachromophore complexes of delements Pt2+, Pd2+, Rh3+, Cr3+, Co2+, Ni2+, Zn2+ and Cd2+ was presented. On basis of experimental data on electron absorption spectra (EAS) and quantum chemical calculations of EAS of model anion-radical 1,10-phenanthroline and electron-rich 1,10-phenanthroline complexes suggest of long-wave bands in EAS of metal phenanthrocyanines was made. Some general regularities of new metal phenanthrocyanines formation in C(sp2 )H-C(sp2 )H-coupling reactions of coordinated 1,10-phenanthrolines concerned with metal promotion and elementary electron and proton transfer processes in initialization stage of C(sp2 )H-C(sp2 )H-coupling was analyse