Generation of recombinant adenoviruses and lentiviruses expressing angiogenic and neuroprotective factors using Gateway cloning technology

The critical aspect in gene and gene-cell therapy is to find an optimal vector - a carrier of genetic information. Viruses represent a natural biological system for gene transfer into eukaryotic cells. One of the most effective and proven vectors for delivery of recombinant nucleic acids into mammalian cells are adenoviruses and lentiviruses. In this study using the Gateway cloning we have constructed adenoviral and lentiviral vectors encoding angiogenic and neuroprotective factors: various isoforms of vascular endothelial growth factor vegf121, vegf165, vegf189; basic fibroblast growth factor fgf2; glial cell-derived neurotrophic factor gdnf. The efficiency of transduction of HEK293A cell line with generated recombinant viruses and expression of recombinant proteins were confirmed by immunofluorescent analysis

Differential immuno-reactivity to genomic DNA, RNA and mitochondrial DNA is associated with auto-immunity

© 2015 S. Karger AG, Basel. Background: Circulating auto-reactive antibodies are hallmark features of auto-immune diseases, however little is known with respect to the specificity of such bio-markers. In the present study, we investigated the specificity of anti-nucleic acid antibodies in the blood of subjects with systemic lupus erythematosus (SLE) and healthy controls. Methods: Sera from 12 SLE cases and 8 controls were evaluated for immuno-reactivity to purified RNA, DNA and mitochondrial DNA (mtDNA) by enzyme-linked immuno-sorbent assay (ELISA). Results: As expected, immuno-reactivity to total nucleic acids was significantly higher in subjects with SLE when compared to healthy controls, however a clear distinction was observed among the various nucleic acid sub-types, with sera from SLE subjects displaying the greatest immuno-reactivity to RNA followed by mtDNA and then total DNA. Conclusion: The identification of auto-reactive antibodies can serve as highly sensitive biomarkers, although their specificity may not always allow diagnostic certainty. The knowledge that auto-antibodies in subjects with SLE display differential immuno-reactivity may help to improve existing diagnostics and may lead to a better understanding of the pathogenesis of auto-immune disorders

Analysis of recombinant VEGF gene expression by genetically modified umbilical cord blood mononuclear cells in experiment in vivo

To obtain a significant therapeutic effect transplanted genetically modified cells should have an enhanced ability to survive and active expression of the therapeutic gene. In this paper, by using immunofluorescent staining we investigated the functional activity of the gene-cell formulation designed to deliver a therapeutic gene into the area of regeneration. As a model we used transgenic SOD1-G93A mice with amyotrophic lateral sclerosis phenotype which received xenotransplantation of human umbilical cord blood mononuclear cells, genetically modified with adenoviral expression vector encoding vascular endothelial growth factor (VEGF) and the reporter green fluorescent protein (EGFP). Results of the study allowed to establish not only the duration of survival of transplanted cells, but also the efficiency of expression of recombinant genes in genetically modified cells in vivo. Double immunofluorescent staining with antibodies against human nuclear antigen HNA and VEGF detected HNA+/VEGF+ cells in the terminal stage of disease 15 weeks after transplantation. These data suggest that genetically modified umbilical cord blood mononuclear cells, transplanted into SOD1-G93A transgenic mice, are able to penetrate the blood-brain barrier and migrate into the area of degeneration of nerve tissue and survive from the time of transplantation until the death of animals at the terminal stage of disease. At that time adenoviral expression vector encoding therapeutic gene is functionally active in transplanted cells, and secretory products of recombinant gene act on target cells by a paracrine mechanism

Региональные особенности современных изменений зимней аккумуляции снега на севере Евразии по данным наблюдений, реанализа и спутниковых измерений

The paper presents data on winter snow accumulation on the Russian territory obtained from different sources: ground measurements of snow depth on meteorological stations, water snow equivalent calculated from data of route surveys, reanalysis, and satellite observations. Three large regions were isolated which are distinguished by characters of the snow depth fluctuations and their long-term tendencies. Comparison of the trends calculated from data of reanalysis and satellite measurements demonstrated essential disagreement of them with data of in situ observations.Рассматриваются данные о зимней аккумуляции снега на территории России из разных источников: наземных наблюдений за толщиной снежного покрова на метеостанциях; водного эквивалента снега по данным маршрутной съёмки, а также реанализа и спутниковых измерений. Выделено три крупных района, отличающихся по характеру колебаний толщины снега и их многолетним тенденциям. Сравнение этих трендов, полученных по данным реанализа и спутниковых измерений, указывает на их существенные расхождения с данными наблюдений

Comparative assessment of physicians’ and senior medical students’ basic knowledge in treatment of chronic obstructive pulmonary disease

Chronic obstructive pulmonary disease (COPD) is one of the most common pathologies of the respira-tory system. This disease ranks third in the group of the main causes of death in the world. The effective treatment of COPD has been developed by today. However, a significant part of physicians has an insufficient amount of education in this matter

Stability of snow cover on the territory of Russia in relation to climate change

The change of the snow cover stability characteristics over the Russian territory including regional and seasonal features of this change in the modern warming period of 1991–2008 in comparison with the period 1961–1990 are investigated in the paper using the data of 223 meteorological stations. Features of the snow cover regime related to teleconnection indices change of the North Atlantic Oscillation (NAO) and the West Pacific (WP) one are analyzed.Opposite tendencies of changes of the snow period duration were observed in most regions of Russia in each of the decades (1991–2008) in comparison with 1961–1990. Differently directed regional changes of the number of days without snow and interruption in snow cover were identified for each of the seasons in periods of intense warming in recent years compared to the baseline thirty years. Similar tendencies of seasonal changes of the number of days with snow depth more than 50 cm were observed in two decades in comparison with 1961–1990. The largest changes occurred in the winter and spring seasons. The spatial distribution of changes of the number of days with snow depth more than 50 cm also demonstrates the same tendency in the years with positive phase of the NAO and WP compared with the years of negative phase

Generation of recombinant adenoviruses and lentiviruses expressing angiogenic and neuroprotective factors using Gateway cloning technology

The critical aspect in gene and gene-cell therapy is to find an optimal vector - a carrier of genetic information. Viruses represent a natural biological system for gene transfer into eukaryotic cells. One of the most effective and proven vectors for delivery of recombinant nucleic acids into mammalian cells are adenoviruses and lentiviruses. In this study using the Gateway cloning we have constructed adenoviral and lentiviral vectors encoding angiogenic and neuroprotective factors: various isoforms of vascular endothelial growth factor vegf121, vegf165, vegf189; basic fibroblast growth factor fgf2; glial cell-derived neurotrophic factor gdnf. The efficiency of transduction of HEK293A cell line with generated recombinant viruses and expression of recombinant proteins were confirmed by immunofluorescent analysis