Gene and cell therapy are tightly associated and quicklydeveloping areas of biomedicine, which purpose is developmentof methods to cure diseases caused by genetic defects and/ordeath of certain cell types. Current state of methods of retinaldiseases gene and cell therapy is analyzed in this review. Wereviewed development of gene therapeutic approaches totreatment of the 2-nd form of Lebers congenital amaurosisand other retinal diseases. We also compared therapeuticpotential of pluripotent stem cells and human adult retinalstem cells. Therapeutic potential of pluripotent stem cellsseems to be better due to their ability for unlimited expansionand organogenesis.

The possibility of hepatocytes differentiation frompancreatic cells is discussed in the review. Both liverand pancreas develop from endoderm so there could be acommon stem cell giving rise to both pancreatic and livercells. Different experimental models are used to studythe possibility of hepatocytes development in pancreassuch as influence of peroxisomes proliferation stimulators,hyperexpression of keratinocyte growth factor in pancreaticislets, Cu depletion-repletion model, etc.). There is noenough data to confirm which particular pancreatic cell typecan be the source of hepatocytes.Literature data allow supposing that hepatocytes canarise in pancreas from three different sources: acinar,endocrine or ductular cells.

Dysferlinopathies belong to neuromuscular diseasesassociated with aberrant expression and/or function ofdysferlin protein in skeletal muscle, which is caused bymutations in the dysf (dystrophy-associated fer-1-like, DYSF)gene. Because of the large size of the codon-optimized dysfcoding region (6243 bp), adenoviral vectors are suitable forthe creation of genetic constructs, which are capable ofdelivering a large amount of recombinant genetic informationinto both dividing and non-dividing cells, as well as provide ahigh level of transgene expression.We generated a recombinant adenovirus serotype5 encoding a codon-optimized gene for human dysferlin (Ad5-Dysf) and analysed recombinant protein expression in vitro inHEK-293T cell line.

Biodistribution of lipoplexes formed of cholenim substancesI-III, containing one, two or three cholesterol moieties, andeukaryotic 14С-DNA and(or) reporter gene into mice organsusing a variety of administration routes (intraperitoneally,i.p.; portal vein or left renal artery) is studied in this paper. Itis shown that biodistribution doesnt depend on lipoplex lipidcomposition under i.p. administration, and depends on lipidnature under vein and artery administration. Effective in vivotransfection and reporter gene expression are demonstratedunder portal vein administration of lipoplex formed ofdicholenim II and lactosylated lipid IV (1 to 1 mass ratio). In thecase, the β-Gal gene expression (above 0,3 mcg / g of tissue)is demonstrated in lungs, liver and spleen histochemicallyand spectrophotometrically. Introduction of cholesterolmoieties into oligoethylenimine structure results in optimalhydrophilicity/hydrophobicity ratio, their stabilization, andoptimal value of critical constant of micelle formation. Thereare certain outlooks due to usage of the lipoplexes describedfor targeted gene delivery.

Evaluation of treatment results of chronic liver diseasesshould be made on the basis of morphological analysis ofliver biopsies. The aim of our study was to investigate theeffect of autologous hematopoietic stem cell transplantationon histology activity index and grade of fibrosis in alcoholicliver cirrhosis patients. The study was performed on liverbiopsies of 11 patients with alcoholic liver cirrhosis. Biopsieswere taken before the injection of autologous peripheralblood stem cells into celiac trunk, 3 and 12 months afterthe procedure. Liver biopsy specimens were stained withhematoxylin-eosin and Van Gieson's. Results showedimprovement of liver structure and decrease in histologyactivity index in liver biopsies performed 3 and 12 monthsafter transplantation. Our data suggest that autologoustransplantation of hematopoietic stem cell in patients withalcoholic liver cirrhosis is effective method that is capableto reduce inflammation activity in the liver, improve itsstructure and decrease liver fibrogenesis.

Atherosclerosis is one of the leading causes of disabilityand death worldwide. Liver plays a huge role in pathogenesisof atherogenic dislipidemia, development and progression ofatherosclerotic lesions. We studied the effect of atherogenicdiet on liver morphology in animal model of diet-inducedatherosclerosis in mice Mus musculus C57BI6. This strainhas a natural ability to develop atherosclerosis, while someother mouse stains has not. After 14 weeks on atherogenicdiet a severe hepathomegaly (9% of body mass) and lobularstructure deformation was found. We also observed signs ofmicro- and macrovesicular steatosis, cell apoptosis, fibrosisand inflammatory leukocyte infiltration. So, liver not only playsan important role in dislipidemia, but it is also a target-organin lipid metabolism imbalance.

Interaction and aggregation of cholesterol and sodiumdodecyl sulfate molecules were studied in this paper.Sodium dodecyl sulfate was taken as a model for biologicalmembranes. Cholesterol-sodium dodecyl sulfate complex wasdescribed by modern methods of nuclear magnetic resonancespectroscopy.Nuclear magnetic resonance spectra were recorded on«Avance-500» spectrometer (Bruker).To assign 1Н signals of cholesterol, sodium dodecylsulfate and cholesterol+sodium dodecyl sulfate mixturein nuclear magnetic resonance spectra literature datawas used, and 2D homo- end hetero-correlation nuclearmagnetic resonance spectra were recorded. To study theformation of sodium dodecyl sulfate micelles and complexof cholesterol-sodium dodecyl sulfate micelles selectivenuclear Overhauser effect spectroscopy experiments werecarried out.The formation of sodium dodecyl sulfate micelles in dimethylsulfoxide solution was confirmed by nuclear Overhauser effectspectroscopy data. The presence of a complex between sodiumdodecyl sulfate micelles and cholesterol molecules has beenproven by selective nuclear Overhauser effect spectroscopyexperiments. Nuclear Overhauser effect between OHgroupof cholesterol and «tail» groups of sodium dodecylsulfate hydrophobic part was observed in the experiment.This observation corresponds to close spatial arrangementof these parts of different molecules and the presence ofa complex between cholesterol and sodium dodecyl sulfatemicelles.On the basis of the nuclear magnetic resonanceexperiments was established that molecules of sodiumdodecyl sulfate form micelles in dimethyl sulfoxide solutionat concentrations above the critical micelle concentration.Cholesterol molecules form an intermolecular complex withsodium dodecyl sulfate micelles by interaction of the OH groupof cholesterol and СН3-1 and СН2-2 «tail» aliphatic groupsof sodium dodecyl sulfate. This interaction is similar to thebehavior of cholesterol in phospholipid bilayer membranes inwhich cholesterol enters its cyclic part in the hydrophobictails of phospholipid molecules oriented primarily across thebilayers.

In medical practice is increasingly use proteolytic enzymesof microorganisms. Particular attention of researchers isattracted proteases, which have fibrinolytic properties,and can lyse clots. Previously had been isolated and purifiedto homogeneity glutamyl endopeptidase, subtilisin-likeproteinase and metalloendopeptidase of Bacillus pumilis3-19, secreted by B. subtilis JB 2036 recombinant strain.The analysis of thrombolytic, fibrinolytic, and anticoagulantproperties of the recombinant enzymes was conducted.It is shown that all investigated proteinases are ableto efficiently lyse the clot. In an in vitro subtilisin andglutamyl endopeptidase of B.subtilis recombinant strainhave anticoagulant activity. Metalloproteinase is notable to influence the process clot formation. Subtilisinlikeproteinase and glutamyl endopeptidase possessfibrinolytic activity and the ability of the activator relativeto plasminogen. Metalloproteinase shows no fibrinolyticproperties. Due to the high incidence of cardiovasculardiseases is urgent search for new enzymes with highbiological activity, specificity and low toxicity.

Here we report the synthesis of (poly)allylamine-coatedsuperparamagnetic iron oxide nanoparticles for the surfacemodification of living cells. Magnetic functionalisation of cowembryonic lung cells did not affect the viability of the coatedcells, confluent monolayer formation and proliferation, asdemonstrated using fluorescence and white light microscopy,flow cytometry and MTT assay. Functionalised cells weremagnetically responsive. We believe that the single-stepapproach described here is a novel and potentially promisingway to functionalise mammal cells with magnetic nanoparticlesfor the subsequent applications in cell therapy, directed cellsdelivery and spatial positioning in tissue engineering.

The lipid vesicles of bisamphiphiles cardiolipin-like dicationiclipids (CDL) I-IV were studied for creation of lipoplexes withplasmid DNA of different sizes to obtain stable lipoplexes forgene transfer to gene therapy. Lipoplexes sizes (300±100 nm)and stablity (> 2 hrs) of CDL were sufficient to be used ingene transfer against monolayer and suspension cell cultures.The CDL total cytotoxicity determined by MTT-test was lowercompare to lipofectin as a control. Transfection conditionsagainst tumor cells lines were optimized by lipoplexes of CDLand plasmid DNA. The most efficient transfection for lipoplexesCDL-plasmid DNA was at the lipid-DNA (L/D) ratio equal to 5(for lipofectin, it was 2). For monolayer cell cultures, lipoplexesCDL-I are comparable in terms of transfection efficacy withlipofectin; in the case of suspension culture, their efficiencywas lower by one order of magnitude. It permits a usageof lipoplexes suggested as mediators for gene transfer anddelivery to human tumor cells.

Cytotoxic ribonucleases (RNAses) are known to beperspective drugs for cancer therapy. The extension of modelobjects range will give possibility to assess the cytotoxicRNAses selectivity.We estimated the effect of Bacillus intermediusribonuclease (binase) and bovine RNAse A to E. coli K 12 andlung epithelia of cow embryo (LEC) cells. LEC cells apoptosiswas characterized with a double staining with annexin-FITC andpropidium iodide (PI). E. coli K12 cells vitality was estimatevia PI staning.Binase and RNAse A were not cytotoxic to LEC andE. coli K12 cells in investigated concentrations (100 and300 ƒg/ml).Low RNAses toxicity for E. coli allows to suppose thebinase and RNAse A in concentrations, capable to displayantitumor activity, will not to effect the tissues microbialflora during in vivo tests. The lack of apoptosis inducingactivity of binase for LEC cells confirms this RNAse selectivityfor tumor cells.

The data obtained on cell lines MDCK showed that forimplementation of the biological function of antibodies to DNAboth normal and pathological need the participation as Fab- andFc-fragments of molecules of IgG. However, antigen-bindingregion is responsible for the manifestation of the biologicalfunction of SLE antibodies to DNA more immunoreactivethan antibodies in normal and constant region is probablyresponsible for the active conformation of antibody for pathobiologicalfunction of antibodies to DNA in the cell.

Nowadays ability to use hematopoietic stem cells fortreatment of various diseases, including kidney pathology,is widely investigated. Umbilical cord blood as the source ofhematopoietic stem cells becomes more and more promising.The purpose of our investigation was to study homing andways of human umbilical cord blood mononuclear cellsdifferentiation in an intact rat kidney. We transplanted humanumbilical cord blood mononuclear cells fraction, enrich withhematopoietic stem cells, into the tail vein of rats. On 2, 5,7 and 14 days after transplantation paraffin kidney sliceswere immunohistochemically stained with antibodies againsthuman leukocyte antigen (HLA-ABC) to study migration anddifferentiation of transplanted cells. Results: HLA-ABC+-cells were revealed in the epithelium of the distal tubule at allexperimental dates. But HLA-ABC was expressed not in eachdistal tubule and not by all tubular cells. We concluded thathuman umbilical cord blood mononuclear cells transplanted intosystemic circulation of rat migrate into the intact kidney andbuilt in the distal tubule epithelium. This data allow to suggestthat distal tubule are stem cell «niche» in the kidney.

Some RNases including ones of microbial origin possessantitumor activity, which mechanisms remains unclear. Herewe investigated the first step of RNase action towardseukaryotic cells which is connected with increase of cellpermeability for ions and macromolecules.Using radiological analysis of 45Са2+uptake by Candidayeast and fluorescence imaging of human embryo kidneycells HEK stained by Ca2+-specific Fura-2/АМ day the levelof intracellular Ca2+ under treatment with the RNase ofBacillus intermedius (binase) was studied. Viability of lungcarcinoma epithelial cells A549 treated by binase wasmeasured by WST proliferation kit, stability of erythrocyteswas tested by lysis assay.We have shown that binase induces the permeabilityincrease of lower and higher eukaryotic cells for Ca2+aswell as the increase of protein permeability of A549 cells.Binase treatment protects erythrocytes from osmoticshock.The protective or cytotoxic binase effect followed byincrease of cellular permeability is realized depending on thedell type, where the expression of КСa channels and of certainoncogens, particularly of ras family, is crucial. The obtaineddata supports the significance of the cell permeability increaseas a primary step in the mechanisms of binase-inducedbiological effects.

One of the most common markers of pancreas stemcells is a stem cell factor receptor C-kit. According to someauthors, this marker is presented in islet cells of normal ratpancreas. But it is unknown about the behavior of these cells indisorders of carbohydrate metabolism during liver disease. Theaim of our study was to evaluate C-kit expression in pancreasafter partial hepatectomy in rats. Partial hepatectomy wasperformed for 27 white male rats. The expression of C-kit,insulin and glucagon in rats pancreas was studied. Theexpression of C-kit in islets and interstitial cells was shownin results after 3 days of the experiment, and double stainingshowed that these cells can express glucagon. Thus, thereis the activation of C-kit+ progenitor cells in pancreas afterpartial hepatectomy and the beginning of there differentiationto ƒ-cells of Langerhance islets.

Cell therapy of various diseases is one of the mostperspective fields in modern medicine. Adipose derivedautological stem cells can be obtained for therapeuticpurposes. Animal model of human diseases are essential forcell therapy research. However, the most frequently usedlaboratory animals, such as rats and mice, cant suffer thewhole rate of common diseases of modern society. At thesame time Syrian hamsters can provide scientists with anappropriate animal models of these diseases. Nevertheless,we couldnt find any data on hamsters stem cells isolationand their characteristics. In this study we first isolated Syrianhamsters adipose derived stem cells, characterized theirmorphology, features and differential potential in severalways. These cells are much alike multipotent mesenchymalstromal cells and can go through osteogenic and, adipogenicdifferentiation. We have also shown that these cells candifferentiate in neurogenic way.

Our research is direct to determination of serineproteases (subtilisin and glutamyl endopeptidase) action atestablish cell lines. We determined that the serine proteasesof bacillus are capable cytotoxic action at establish cell linesof animals. The cell lines are differentiating by sensitivity toproteins. The cell lines LEK and NGUK were more sensitiveto subtilisin, and line Vero - glutamyl endopeptidase.

To increase the viability of neural cells in neurodegenerativediseases, after neurotraumas and ischemic strokes themost important neurotrophic and neuroprotective factors,which can be used as therapeutic agents were identifiedin long-term studies in vitro and in vivo. These includebrain-derived neurotrophic factor (BDNF), glial-derivedneurotrophic factor (GDNF), insulin-like growth factor (IGF)and vascular endothelial growth factor (VEGF). One of thepromising ways of the delivery of supporting neuron survivalfactors is considered to be transplantation of geneticallymodified cells overexpressing recombinant therapeuticgenes. This article describes generation of cellular deliveryvectors of therapeutic genes - human umbilical cord bloodmononuclear cells genetically modified by dual cassetteplasmids, expressing two therapeutic genes. Efficiency oftransgene expression was confirmed in vitro using RT-PCR.Analysis of survival, migration, and phenotype of geneticallymodified cells was performed 2 weeks after transplantationinto transgenic mice with amyotrophic lateral sclerosisphenotype.

Introduction. Recent studies certify the existence of linkbetween Alzheimers disease and cardiovascular pathology,however the mechanisms of this phenomenon is unclear. Herewe studied the influence of Alzheimers β-amyloid peptide(βAP) on the contractility of rat myocardium and aorta.Material and methods. Contractility of myocardium ventriclestrips and transverse fragments of abdominal aorta wasmeasured at Power Lab setup using conventional myographictechnique. Contractile responses of aorta strips were evokedby application of receptor agonists, contractile responses ofmyocardium - by electrical stimulation. Contractile responsesof aorta strips after application of carbachol (10-6-10-4 М),histamine (10-6-10-4 М), norepinephrine (10-5-10-3 М) andATP (10-6-10-4 М) were measured.Results and discussion. We found the impairment ofcarbachol- and histamine-induced contractility of aorta,appearing as perverse contractile reactions (relaxation insteadof contraction) under the action of βAP (10-6 М). Next, wefound βAP-induced impairments of ventricle myocardiumcontractility, appearing as decrease of relaxation phaseduration and increase of relaxation speed (positive lusitropiceffect). Also, own positive lusitropic effect of norepinephrinewas absent in presence of βAP (10-6М).Thus, βAP(25-35) significantly impairs the contractilityof rat myocardium and aorta, as well as processes of itsregulation. Obtained data significantly broad our understandingof mechanisms of Alzheimers disease pathogenesis andpathophysiology of cardiovascular system.

The work is aimed at the study of the effect of novelblock copolymers of ethylene oxide and propylene oxideon the delivery of plasmid DNA and its complexes withcationic polymers into human cells. Tri-functional amphiphilicblock copolymers on the basis of glycerol (LaprolsTM),polyethyleneimine (25 kDa) and commercial transfectionreagent TurboFectTM were tested as delivery systems.TurboFect was found to form more compact and positivelycharged polyplexes with plasmid DNA (pEGFP-N2) andprovided higher expression of GFP in HEK 293 cellscompared to polyethyleneimine. Laprols weakly interactedwith plasmid DNA and did not improve its intracellulardelivery. However they markedly promoted cell transfectionby DNA-polyethyleneimine complexes. Results show thatLaprols exhibit mild cytotoxicity and produce the interest forgene therapeutics delivery into cells.

To determine the antiviral activity of various biologicallyactive compounds, the model of adenovirus infection on thebasis of cell cultures of human HEK293A and recombinantadenovirus Ad-EGFP, expressing green fluorescent proteinEGFP. Adenoviruses have a capsid size of 70-90 nm and areable to infect dividing and nondividing cells in vitro and in vivo.Recombinant adenoviruses are the replicative defect in thecells of humans and animals. The developed model allowedus to determine the effect of bacterial proteases in theinfected cell cultures with adenovirus. This model can alsobe used for screening drugs with potential protivivovirusnoyactivity.

Effect of binase (RNAse of Bacillus intermedius) onphorbol myristate acetate-(PMA)-induced apoptosis of humanperipheral blood granulocytes and monocytes was studied invitro by flow cytometry. Both toxic (400 μg/ml) and nontoxic(40 μg/ml) binase concentrations were tested. The binaseend-point effect was dependent on the target cell populationand the binase concentration. In a granulocyte subset, the400 μg/ml concentration resulted in strongly pronouncedstimulation of PMA-induced apoptosis. In a monocyte subset,the 40 μg/ml concentration developed a protective effect asjudged by an increase in a percantage of viable cell subset andby slowing-down cells transtion from an early to late PMAinducedapoptotic phase.

During liver fibrosis development connective tissueis produced by myofibroblasts that could originate fromtwo hepatic populations: hepatic stellate cells and portalfibroblasts. A marker of myofibroblasts is the expressionof ƒ-smooth muscle actin (ƒ-SMA). Distinctive feature ofmyofibroblasts, derived from hepatic stellate cells, is thepreservation of the hepatic stellate cells marker expression -desmin. The processes of activation, proliferation and cellstrans-differentiation into myofibroblasts are closely relatedto the activity of transcription factor NF-kB and its inhibitorIkBƒ. The aim of our work was to obtain a culture of hepaticmyofibrobasts, to study their origin, phenotype, relationsbetween NF-kB and IkBƒ expression and the processes ofactivation and cells trans-differentiation into myofibroblasts.For this purpose we isolated heterogeneous population ofcells from rat liver by the method of explantation. Almostall the cells had desmin and ƒ-SMA expression. On thisbasis, we suppose that these myofibroblasts were hepaticstellate cells derivatives, and singular desmin-negative cellsoriginated from portal fibroblasts. Thus, hepatic stellate cellshave major potential to activation, growth, proliferation andtransdifferentiation into myofibroblasts in comparison toportal fibroblasts. Activated state of the cells was confirmedby stable expression of NF-kB and its inhibitor IkBƒ in all thecells throughout the whole experiment.

The herpes simplex virus thymidine kinase/gancyclovir(HSV-tk/GCV) system is studied as cytotoxic lipoplex basedon cardiolipin-like dicationic lipid CDL-I. It is proposed to beused as nonviral gene transfer system in cancer gene therapyprotocols. Аn efficient transfection of MCF7 and HEC293 celllines with this lipoplex was earlier demonstrated. Non-viralsystem based on the CDL-I/HSV-tk lipoplex and ganciclovirtreatment causes efficiently death of tumor cells with aninvolvement of apoptosis key stages. It was proved thatdepolarization of mitochondrial membrane and increased levelof NF-kB transcription factor take place as a response to CDL-I/ HSV-tk lipoplex action followed by gancyclovir treatment. Itsuggests an early involvement of apoptosis mitochondrial wayto an action of this certain «suicide» system, delivered usingdicationic lipid.

A deposition of cationic (chitosan) and anionic (alginicacid) polysaccharides onto the surface of normal and cancerhuman cells was studied with the use of dynamic lightscattering. A method for preparation of multilayer polymericshell by means of electrostatic adsorption of polysaccharidesonto cell plasma membrane has been proposed. According toconfocal microscopy, the polymeric shell evenly covers the celland is 1-5 μm in thickness. Under experimental conditions,the modification with polysaccharides inhibits human skinfibroblasts growth but does not exhibit cytotoxicity toHeLa cells. We developed an approach to controllable andreversible aggregation of modified cells by their cross-linkingin the presence of calcium ion. Resulting aggregates havespherical shape and the size of 100-500 μm. Proposedapproaches and methods represent an alternative to cellmicroencapsulation technique and are of interest for thedevelopment of three-dimensional cell models and theirdelivery in vivo.

Schwann cells are a major figure in the process ofregeneration in the peripheral nervous system. They migrateinto the injury region of spinal cord, which are involved inremyelination and are regarded as the source of numerousmolecular signals that could potentially support the growthof axons in the central nervous system. In the present workwe describe the behavior of migrating into the injury dosedregion spinal cord Schwann cells under the influence ofneurotrophic factors - vascular endothelial growth factor(VEGF) and fibroblast growth factor 2 (FGF2), deliveredby direct introduction of «naked» plasmid DNA and bytransplantation of genetically modified human umbilical cordblood mononuclear cells.Using immunohistochemical detection of markers of S100,GFAP, Krox20 and HSP25 identified different phenotypesof migrating into the spinal cord of endogenous Schwanncells. Found that greatest influence on their numbers in theinjury region provides local delivery of genes vegf and fgf2by human umbilical cord blood mononuclear cells. However,the direct introduction of the same plasmid may also bepromising in the case of synthetic platforms that enhanceits transfection activity.

To obtain a significant therapeutic effect transplantedgenetically modified cells should have an enhanced abilityto survive and active expression of the therapeuticgene. In this paper, by using immunofluorescent stainingwe investigated the functional activity of the gene-cellformulation designed to deliver a therapeutic gene into thearea of regeneration. As a model we used transgenic SOD1-G93A mice with amyotrophic lateral sclerosis phenotypewhich received xenotransplantation of human umbilical cordblood mononuclear cells, genetically modified with adenoviralexpression vector encoding vascular endothelial growthfactor (VEGF) and the reporter green fluorescent protein(EGFP).Results of the study allowed to establish not only theduration of survival of transplanted cells, but also theefficiency of expression of recombinant genes in geneticallymodified cells in vivo. Double immunofluorescent stainingwith antibodies against human nuclear antigen HNA andVEGF detected HNA+/VEGF+ cells in the terminal stage ofdisease 15 weeks after transplantation. These data suggestthat genetically modified umbilical cord blood mononuclearcells, transplanted into SOD1-G93A transgenic mice, areable to penetrate the blood-brain barrier and migrate intothe area of degeneration of nerve tissue and survive fromthe time of transplantation until the death of animals at theterminal stage of disease. At that time adenoviral expressionvector encoding therapeutic gene is functionally active intransplanted cells, and secretory products of recombinantgene act on target cells by a paracrine mechanism.

Impairment of axon transport is widespread and earlyevent in a number of neurodegenerative diseases. The goalof study is to investigate the mechanisms of retrograde axontransport impairment in mouse spinal motoneurons afterapplication of ƒ-amyloid peptide (ƒAP) (25-35) on the centralstump of transected sciatic nerve.Retrograde fluorescent tracer Fluorogold (5%), ƒAP(25-35) (10-6 М), or mix was applied to the proximal stumpof the transected sciatic nerve of mouse under the generalanesthesia. At 24 hours after surgery lumbar spinal cordwas processed for morphometric and immunohistochemicalanalysis.The amount of Fluorogold-positive motoneurons at controlwas 1223,7162,7 (n = 7), whereas after application ofƒAP(25-35) - 393,285,3 (n = 5, p < 0,01), which certifiespronounced inhibition of retrograde axonal transport. Stainingwith polyclonal antibodies against caspase-3 did not revealmotoneurons in apoptotic state. Staining with monoclonalantibodies against the ƒAP (25-35) was negative both atoperated and intact sides of spinal cord.Thus, revealed inhibitory action of ƒAP (25-35) on theretrograde axon transport is not related to apoptotic death ofneurons or accumulation of ƒAP (25-35) inside the neuronalsoma, but, evidently, is mediated by intraaxonal effects.Obtained data has great importance for understanding ofmechanisms of Alzheimers disease pathogenesis.

One of the most common markers for stem cells inpancreas is the stem cell factor receptor C-kit (CD117) thatplays a main role in differentiation of progenitor endocrinecells of pancreas islets in prenatal development and persistsafter birth. But still the role of C-kit positive cells in isletƒ-cells regeneration during the diabetes mellitus type I hasnot been studied. Thats why the aim of our work was tostudy the dynamic of C-kit expression in the pancreas isletsduring the experimental alloxan diabetes in rats. The workwas made on 33 rats with the experimental diabetes. Bloodglucose levels, levels of insulin and glucagon were measured.And also we studied the expression of C-kit, insulin andglucagon in rat pancreas. The results of the study showedthe C-kit expression after one day of the experimentalhyperglycemia. These cells were also expressed insulin andglucagon. We suppose that C-kit+-cells, which produceinsulin, were enable to correct disrupted carbohydratemetabolism during alloxan diabetes.

For the successful application of plasmid vectors in genetherapy protocols it is necessary to develop methods forpurification of highly homogeneous preparations of recombinantDNA that do not contain contaminants, primarily chromosomalDNA, bacterial proteins, RNA and endotoxins. In the courseof our study we performed optimization of the purification ofplasmid supercoiled DNA by three chromatographic steps froman alkaline lysate of bacterial strain of E. coli. We determinedan optimal conditions for alkaline lysis step in order to increasethe yield and minimize the duration of the plasmid purificationby gel filtration.

Autophagy is a fundamental process that ensuresthe regulation of T-cell homeostasis. In case of apoptosisinduction disruption in the cell it could be single mechanism ofthe cell death. Previously was shown inhibition of lymphocyteapoptosis in patients with bronchial asthma, so the main studyof this work has focused on the study development processof autophagy in T-lymphocytes of patients with bronchialasthma. The article presents the main morphological changesin cells associated with activation of autophagy (formationautophagosome). In addition to morphological changes inlymphocytes, we have shown the expression of autophagymarker protein (LC3B). We found that in T-lymphocytes ofpatients with severe asthma are simultaneous activation ofboth autophagy and apoptosis, and autophagy is a stimulusto cell death.

Lentiviral vectors are widely used in genetic modificationof human and animal cells (lentiviral transduction) to enhancetheir therapeutic potential by expression of recombinantprotective and trophic factors. Genetic modification of cells invitro or ex vivo achieves the specificity of viral transduction,as modified are just cells that have been manipulated inthe laboratory. In addition, the introduction of geneticallymodified cells, but not pure virus, helps to avoid introductionof viral particles into the body of the recipient. This approachallows us to control the expression of therapeutic genes, theimmunogenicity of viral vectors and viral transduction. Todate, different approaches are used to improve the lentiviraltransduction (polycations, protamine sulfate, etc.), but thesemethods suffer from limited efficacy or high toxicity. For thefirst time we demonstrated that the recombinant histoneN1.3 increases the efficiency of lentiviral transduction bymore than 2 times and has no toxic effect on target cells ina wide range of concentrations studied.

Human stem cells secretome is currently a very hot areaof research. We report that multipotent mesenchymal stromalcells isolated from human third molar dental follicles (MMSCTMDF),are able to secrete high levels of vascular endothelialgrowth factor (VEGF) when cultured in vitro. Due to the factthat VEGF is a well known angiogenic and neuroprotectivefactor, the use of MMSC-TMDF is promising for thedevelopment of stem cell therapy of various degenerativehuman diseases.

We studied the influence of the culture fluid of fungi ofthe genus Trichoderma on Swiss Webster CFW mice afterexposure to pyrene - polycyclic aromatic hydrocarbons, whichcan cause pathological changes in the body. Beneficial effectof Trichoderma metabolites on haematological parameters,the functioning of liver and nephros was shown, the trendtoward regeneration of the structure of skin and liver afterthe damages, caused by the introduction of pyrene, wasidentified.

The critical aspect in gene and gene-cell therapy is tofind an optimal vector - a carrier of genetic information.Viruses represent a natural biological system for genetransfer into eukaryotic cells. One of the most effectiveand proven vectors for delivery of recombinant nucleic acidsinto mammalian cells are adenoviruses and lentiviruses. Inthis study using the Gateway cloning we have constructedadenoviral and lentiviral vectors encoding angiogenic andneuroprotective factors: various isoforms of vascularendothelial growth factor vegf121, vegf165, vegf189; basicfibroblast growth factor fgf2; glial cell-derived neurotrophicfactor gdnf. The efficiency of transduction of HEK293A cellline with generated recombinant viruses and expression ofrecombinant proteins were confirmed by immunofluorescentanalysis.

Hepatic stellate cells are considered as one of the potentialstem cells candidates in the liver. The aim of our work was tostudy the probability of hepatic stellate cells transplantationto rats after partial hepatectomy, their further homing, theways of differentiation and hepatocytes repopulation in therecipient liver. For this reason fresh isolated rat`s hepaticstellate cells were transplanted into portal vein of intact rats(control group) and rats immediately after partial hepatectomy(experimental group). Before transplantation cells werelabeled by adenovirus expressing green fluorescent protein.Our results showed that it was possible to detect 2 types ofdonor cells in the recipient liver of control and experimentalgroups: 1) hepatocyte-like cells in liver parenchyma; 2) small,spindle-shaped, rounded and triangular cells in liver sinusoidsand portal areas. Transplantation after partial hepatectomyleads to significant increase of transplanted cells homing andstimulation of their differentiation into hepatocytes. Overthe whole experiment there was no hepatic stellate cellstransdifferentiation into myofibroblasts, thus there is no risk ofliver fibrosis development after this cell type transplantation.In summary hepatic stellate cells after being transplanted areable to differentiate into hepatocytes and do not induce liverfibrosis, that confirms their role in organ regeneration andprobable belonging to hepatic progenitor cells.

We studied the adsorption of bioadhesive polymers(polyornithine, gelatin, laminin) on polystyrene surfaceby the use of dynamic light scattering. The contributionof biopolymers to resulting zeta potential of the modifiedsurface was assessed. PC12 cells do not exhibit selectiveadhesion in the presence of foetal bovine serum. Polystyrenewith adsorbed polyornithine promotes primary adhesionof PC12 cells cultured in serum-free medium with nervegrowth factor. Subsequently adsorbed laminin inducesspreading and differentiation of the cells into neuronaldirection. Primary neurons isolated from rat spinal ganglionadhere preferentially on the polyornithine-modified surface.On the polyornithine-laminin surface neurons intensivelyform neuritis that correlates with proliferation of glialcells positive for S100 protein. The results show thatPC12 cells and primary neurons exhibit similar response tosurface material with the latter cells being more sensitiveto this factor. Isolated cell culture can be used to study therelationship between neurite outgrowth and Schwann cellsproliferation on different biomaterials.

The article presents a clinical case of successful surgicaltreatment of a patient with idiopathic progressive hemifacialatrophy. We have used lipofilling with autologous fat tissueenriched by stromal-vascular fraction cells.Observation of the patient within two years suggests thatthis method of treatment is safe and highly effective methodof face soft tissue defect substitution of such pathology.

In this paper we present the clinical observation ofsuccessful treatment of distal form of peripheral arterydisease of the lower extremity with symptoms of criticalischemia in a 60 years old patient.Intramuscular injection of dual expression plasmid, encodingvascular endothelial growth factor VEGF and basic fibroblastgrowth factor FGF2, was performed to the affected lowerextremity. The effect of treatment was evaluated by functionaltests: measurement of ankle-brachial index, treadmill test, therecovery time, shoulder-ankle index after strain and temporaryocclusion. Performed immunohistochemical examination ofbiopsy samples of the affected lower extremity muscles.

To date, advances in the field of tissue engineering,cell transplantation and genetic engineering have madethe biological materials of different origin an importanttherapeutic tool in clinical medicine. Currently, cellspreservation is achieved by freezing at -80°С or in liquidnitrogen. Cryopreservation technology is expensive andhas considerable limits during transportation. Preservationof viable biological material in dry state under ambienttemperature is considered as attractive, but yet fullyachieved alternative. There are organisms which are able tosurvive complete water loss. Understanding of mechanismsunderlying dehydration tolerance will allow the developmentof dry preservation technology for molecules, cells andorgans, and further use of these methods in medicine,pharmacology and biotechnology.