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Vol 9, No 3 (2014)

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Open Access Open Access
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To the 90th year of Alexandr Jakovlevich Friedenstein

Zorin V.L., Zorina A.I.


The article is dedicated to the outstanding Russian scientist A.J. Friedenstein, a founder of the hematopoietic organs' stroma doctrine, a discoverer of mesenchymal stem cells (multipotent mesenchymal stromal cells, MMSC), an initiator of the tests to determine the «verity» of stem stromal cells. Discoveries of the scientist and scientific school he created are used both in the research and clinical practice all over the world.
Genes & Cells. 2014;9(3):8-10
pages 8-10 views

Cell technologies for muscle tissue restoration. Part I: myocardium

Korsakov I.N., Zorin V.L., Eremin I.I., Zorina A.I., Kotenko K.V., Kopnin P.B., Pulin A.A.


This review represents ongoing clinical studies devoted to recovery of heart muscle using cell technologies, as well as analysis of applied populations of cells.
Genes & Cells. 2014;9(3):11-17
pages 11-17 views

Chondroitinsulfate proteoglycan receptors in the nervous system

Paveliev M.N., Baltina T.V.


Chondroitinsulfate proteoglycans (CSPG) play a crucial role in the inhibition of posttraumatic axonal regeneration in the central nervous system. Understanding of the mechanism of the CSPG action on axonal regeneration is important for treatment of patients after the brain and spinal cord injuries. Here we make a review of recent studies that discovered CSPG receptors acting in the plasma membrane of neurons.
Genes & Cells. 2014;9(3):18-21
pages 18-21 views

Perspectives of using stem cells in diabetes mellitus treatment

Plushkina A.S., Kaligin M.S.


Using stem cells is one of the most perspective methods of diabetes mellitus treatment. Different stem cells populations are used for this purpose. Pancreatic stem cells are considered to be the most appropriate. This review is devoted to new methods of diabetes mellitus treatment by using stem cells and perspectives of using C-kit- positive pancreas cells and desmin-positive stellate pancreas cells as the main candidate to the role of pancreatic stem cells.
Genes & Cells. 2014;9(3):22-24
pages 22-24 views

Halloysite nanotubes is a promising biocompatible material for «smart» composites with encapsulation of biologically active substances

Rozhina E.V., Danilushkina A.A., Naumenko E.A., Lvov U.M., Fahrullin R.F.


Halloysite clay mineral is found in kaolin deposits and is a multilayered nanotube with external diameter of 50 nm inner diameter 15 nm and length1-2 micron. Halloysite is a rolled kaolin and it has found applications in fine porcelain, polymeric composites, tissue engineering, antimicrobial coatings and in cosmetics. Halloysite is also applied as microvesicles to carry drugs and enzymes. It is used as an additive to improve the mechanical strength of polymers. Halloysite is non-toxic to living organisms and the environmentally safe. This review summarizes recent data on the structure of halloysite and its applications.
Genes & Cells. 2014;9(3):25-28
pages 25-28 views

Peribiliary glands of biliary tree as a niche of multipotent stem cells

Sharipova E.I., Gazizov I.M., Gumerova A.A., Kiassov A.P.


Peribiliary glands are located along the large bile ducts. They are found in human and majority of the animals. Other than mucous production, their function had not been defined until recently. But nowadays the question of their multiple functions is actively studied. Due to the last years reports peribiliary glands contain multipotent stem cells, which can differentiate into hepatocytes, cholangiocytes or pancreatic islets cells. The structure and function of peribiliary glands, known experimental models and perspectives of peribiliary glands use in regenerative medicine are discussed in the review.
Genes & Cells. 2014;9(3):29-33
pages 29-33 views

Physiological role of hydrogen sulfide in nervous system

Yakovlev A.V., Sitdikova G.F.


The review provides modern data and the results of author's research on physiological and pathological roles of the new gasotransmitter - hydrogen sulfide (H2S) in the central and peripheral nervous system. H2S is synthesized by three enzymes: cystathionine p-syntase, cystathionine y-lyase and 3-mercaptopiruvat sulftransferase/cysteine aminotransferase. In nerve systems the main source of synthesis H2S is cystathionine p-syntase and high level enzyme expression observed in the embryonic and early postnatal period of organism development that is apparently necessary for the growth and maturation of neural networks for the protection of neurons and astrocytes in the conditions of oxidative stress. Cystathionine p-syntase gene mutation in humans leads to an autosomal recessive metabolic diseases, mental dysfunction, vascular lesions and hyperhomocysteinemia. The aim of this review is to present the currents data about the effects of H2S on ion channels, transmitter release, its participation in the pathology of various neurodegenerative diseases, as well as its antioxidative and neuroprotective action in central and peripheral nervous systems.
Genes & Cells. 2014;9(3):34-40
pages 34-40 views

The role of pancreatic stellate cells in pancreas regeneration in copper-deficient diet model in rats

Abdulkhakova A.R., Galyavieva A.R., Abdulkhakov S.R., Trondin A.A., Pevnev G.O., Titova M.A., Gumerova A.A., Kiassov A.P.


The possibility of pancreatic cells transdifferentiation to hepatocyte-like cells in rat copper depletion-repletion model has been shown in a number of articles. The aim of our research was to study the role of pancreatic stellate cells in pancreas regeneration in the above mentioned model. 24 male Wistar rats were fed a copper-deficient diet, containing triethylenetetramine tetrahydrochloride, for a period of 8 weeks, then animals were returned to the normal rat chow for up to a period of 8 weeks. Animals were killed at the end of 4, 6, 8 weeks of copper-deficient diet and 4, 6, 8 weeks after they were returned to normal diet. Paraffin-embedded sections of pancreas were stained immunohistochemically with antibodies to desmin, the pancreatic stellate cells marker. The acinar tissue structure destruction was observed starting from the period of 4 weeks of copper-deficient diet. Partial acinar restoration was observed 6 and 8 weeks after rats were returned to the normal balanced rat chaw. An increase in the number of desmin-positive stellate cells both in pancreatic acinar tissue and in islets was observed at all periods of experiment. Along with the partial acinar tissue recovery desmin-positive cells were found mainly in areas of newly formed pancreatic tissue.
Genes & Cells. 2014;9(3):41-44
pages 41-44 views

Elimination of PKH26-labeled MMSC after allogeneic transplantation

Arutyunyan I.V., Elchaninov A.V., Fatkhudinov T.H., Makarov A.V., Kananykhina E.Y., Bolshakova G.B., Glinkina V.V., Goldshtein D.V., Sukhikh G.T.


The transplanted allogeneic multipotent mesenchymal stromal cells (MMSC) were previously thought to be poorly recognized by host immune system; the prolonged survival of these cells in host tissues was explained by their privileged immune status. As long as this concept is currently being revised, the understanding of MMSC routes should be reconsidered given the emerging role of host immune system in their gradual elimination. The study was focused upon elimination of PKH26-labeled MMSC, derived from umbilical cord, analyzed in animal models for two distinct pathologies: subtotal liver resection and critical skeletal muscle ischemia. Specific patterns of PKH26-positive macrophages (defined as CD68+ cells) were described for intact spleen and regenerating liver, and for the ischemic skeletal muscle, respectively. The PKH26-positive cells were observed in spleen of the subtotally hepatectomized model animals at 24 h. after surgery combined with MMSC transplantation; 83,2±4,6% of these were CD68+; the ratio reached 100% 3 days after transplantation. The PKH26-positive cells were also detected in regenerating liver starting from 3 days after transplantation, the great majority of them were CD68+ (96,8±2,2% and 96,3±2,6% for 3 and 10 days after transplantation, respectively). A different sort of host environment was provided by the damaged skeletal muscle model: productive phase of aseptic inflammation triggered by ischemia. The PKH26-positive fraction in the pool of macrophages significantly increased from 48,1 ±3,2% 3 days to 76,2±3,9% 30 days after transplantation. Thus, transplanted allogeneic MMSC are recognized and eliminated by host immune system. The rates of elimination depend on site of injection and time elapsed since the injection; the efficacy may reach 100%. The presence of РКН26 vital label (as well as any other exogenous label) in living cell can by no means solely prove its exogenous origin. The massive elimination of MMSC by host macrophages leads to impregnation of the latter with the dye that is masking the true presence of the former. The study accentuates the need of additional criteria for correct data interpretation.
Genes & Cells. 2014;9(3):45-52
pages 45-52 views

Influence of liver perisinusoidal cell transplantation on liver regeneration after damage induced by CCL4 and 2-acetylaminofluorene

Burganova G.R., Titova A.A., Sharipova E.I., Pevnev G.O., Mavlikeev M.O., Gazizov I.M., Galyavieva A.R., Shafigullina A.K., Kaligin M.S., Titova M.A., Gumerova A.A., Kiassov A.P.


Research in the past decade allows us to consider liver perisinusoidal cells as a possible regional stem cells of the liver. Despite of this, there is no work dedicated to study their influence to liver regeneration. The aim of this work was to study liver regeneration after liver perisinusoidal cell transplantation in rats with CCL4 and combination of CCL4 and 2-acetylaminofluorene damage. Our results showed that in both models of toxic damage transplanted perisinusoidal cells are found in the recipient's liver and exhibit the properties of stem cells by differentiation into hepatocytes. Transdifferentiation of injected cells into myofibroblasts was not detected which indicates the absence of liver fibrosis risk after use of this cell type for cell therapy.
Genes & Cells. 2014;9(3):53-58
pages 53-58 views

Application of genetically modified umbilical cord blood cells and direct gene therapy for treatment of skin wounds in rats

Gazizov I.M., Salafutdinov I.I., Mavlikeev M.O., Bashirov F.V., Islamov R.R., Rizvanov A.A., Kiassov A.P.


There were proposed several methods for stimulation of skin wound repair over the last few decades. The most perspective among them are gene and stem cell therapy. In our experiments we combined both approaches by application of human cord blood mononuclear cells (hUCB-MC) transfected with pBud-VEGF165-FGF2 plasmid to enhance healing of full thickness skin wounds in rats. Dual expression cassette plasmid pBud-VEGF165-FGF2 encodes both VEGF and FGF2 therapeutic genes, expressing pro-angiogenic growth factors. Our results showed that in 2 weeks after transplantation some transplanted cells still retain expression of stem cell and hematopoietic markers C-kit and CD34. Other transplanted cells could be found among keratinocytes, hair follicle cells, endothelial cells and in derma. Study of PCNA expression revealed that application of transfected cells terminate proliferative processes in regenerating wound earlier then application of untransfected cells.
Genes & Cells. 2014;9(3):59-62
pages 59-62 views

Transformed lung epithelial cells produce microbial RNAse (binase)

Dudkina E.V., Singh I., Ulyanova V.V., Shah Mahmud R., Barreto G., Ilinskaya O.N.


Today, the targeted drug delivery to tumors is one of the leading researches in anticancer therapy. Using gene engineering techniques the new expression system based on pCS2+ vector was created. Gene of Bacillus pumilis ribonuclease - binase, and gene of intracellular inhibitor of Bacillus amyloliquefaciens - barstar were cloned under promotor of cytomegalovirus. The possibility of binase's gene expression in transformed murine lung epithelial cells was shown. The new plasmid system provides selective cytotoxic action of binase against target tumor cells due to high expression level of foreign protein in transformed cells.
Genes & Cells. 2014;9(3):63-67
pages 63-67 views

Isolation, analysis and application of authogenic adipose derived multipotential mesenchymal stromal cells from dog for therapy pseudoarthrosis of tibial bone

Zakirova E.Y., Zhuravleva M.N., Masgutov R.F., Usmanov R.A., Rizvanov A.A.


This study demonstrates the possibility of using cell-based therapies for the treatment of the longterm intergrows fractures and pseudoarthrosis in dogs. Multipotent mesenchymal stem cells (MMSCs) were isolated from adipose tissue of an adult dog. Multipotent cells was confirmed by the ability to differentiate into 3 main directions (osteogenic, adipogenic and chondrogenic), as well as the expression of the markers CD73, CD105, vimentin. Clinical application received MMSCs in the treatment of pseudoarthrosis of tibia in dogs stimulated bone formation processes. The result of cell therapy was callus formation and recovery of the support function of the injured extremity.
Genes & Cells. 2014;9(3):70-75
pages 70-75 views

Oral mucosais a new source for myoblast derivation

Zorin V.L., Eremin I.I., Rybko V.A., Zorina A.I., Kotenko K.V., Pulin A.A., Kopnin P.B.


Leading laboratories in the world intensively research tissue-specific stem cells. The main goal of such work is development of improved technique of tissue regeneration stimulation. Optimization of conditions of differentiation induction, and search for new sources of cells are actual problems in this field of science. Human gingival mucosa is one of promising sources of cells. Gingival mucosa-derived multipotent mesenchymal stromal cells (gm MMSC) have a number of features, notably the high proliferative activity and ability to multilineage differentiation. However, their myogenic differentiation has not been proofed yet. The study was conducted on 10 gm MMSC cultures obtained from gingival mucosa biopsy samples of 10 healthy volunteers. Possibility of gm MMSC obtainment with high proliferative potential and ability of cells to efficiently differentiate not only into orthodox (adipogenic, osteogenic, chondrogenic) directions but also in myogenic direction during both early and late passages was demonstrated for the first time in our work. Our results of gm MMSC investigation and characteristics of the cell'ssource, confirm advantages of gm MMSC use in regenerative medicine, in particular for the treatment of the muscle tissue diseases of different etiology.
Genes & Cells. 2014;9(3):76-84
pages 76-84 views

Proliferation of cells of the pancreas in experimental diabetes

Kaligin M.S., Titova A.A., Plushkina A.S., Titova M.A., Gumerova A.A., Kiassov A.P.


The proliferative process intensification in the disrupted organs is very important in reparative regeneration. But the possibility of pancreas cells proliferation during damage is not been studied. The aim of our work was to examine the proliferation activity of pancreas during the experimental alloxan diabetes. The work was made on 33 rats. Blood glucose levels were measured. Also the expression of nuclear antigen of proliferating cells PCNA was studied in pancreas cells. After the day of the experimental hyperglycemia PCNA expression in islet and acinar cells of pancreas was found, which was persisted at all stages of the experiment. After 2 days of the experiment single PCNA-positive cells with the cytoplasmic staining were found in the interstitium, near vessels and ducts. Proliferating cells, which produce insulin were found in islets during double staining. Thus, there is the proliferation in islet and acinar cells of pancreas during experimental diabetes type I after alloxan damage. Also, the results of our study confirms the ability of p-cells and progenitor pancreas cells proliferation.
Genes & Cells. 2014;9(3):85-88
pages 85-88 views

Statistical laws of eukaryotic DNA patchiness

Kayumov A.R., Saetgaraeva A.A., Markelov O.A., Bogachev M.I.


Active development of the genetic engineering and expression of foreign genes in various organisms revealed the requirement of the DNA sequences adaptation to the genetic machinery of a host cell including adaptation of both functional elements of the genetic code and its tertiary architecture. The annotated genomes of organisms at different evolutionary levels that are widely used as models were obtained from the Genbank ( The probability density functions of the sizes of structural elements of the genetic code were assessed and analyzed. The analysis of the distribution of sequences of coding DNA (genes and exons) and noncoding DNA (intergenic sequences and introns) revealed their universal pattern in genomes of all eukaryotes independently of their evolutionary level, the average number of introns in a gene, their sizes and the total genome size. It allows claiming that mechanisms of genomic reorganizations as a result of insertions, deletions, mutagenesis, duplications and others exhibit universal character. The size of inserted/deleted sites of DNA directly depends on the average size of the respective structural elements of a genetic code (genes, introns, exons) of the organism. Therefore the genetic engineering designs where the DNA donor and recipient are located at different evolutionary levels require the structural elements of foreign DNA being adapted to their average sizes of the host to minimize the negative effects from the reorganization of the genetic machinery of the host.
Genes & Cells. 2014;9(3):89-93
pages 89-93 views

Role of potassium channels in the negative inotropic effect of hydrogen sulfide in mouse atrium

Lifanova A.S., Khaertdinov N.N., Zakharov A.V., Gizzatullin A.R., Sitdikova G.F.


The effect of sodium hydrosulfide (NaHS) - donor of hydrogen sulfide (H2S) on the force of contraction of isolated mouse atrium was studied. Cumulative application of NaHS in concentrations 100, 200 and 300 ^M induced dose-dependent decrease of the force of contraction, the maximum velocity of contraction and relaxation of the myocardium. A substrate of H2S synthesis - L-cysteine in concentrations 1, 10, 50 uM also had the negative inotropic action, whereas a blocker of H2S synthesis - p-cyano alanine caused an increase of the force of contraction. Inhibition of K-channels by tetraethylammonium (2 mM) caused the increase of the amplitude of contraction and the reduction of negative inotropic effect of NaHS in all used concentrations. After the inhibition of ATP-dependent K-channels by glibenclamide NaHS action was prevented in concentration 100 uM, significantly decreased in concentration 200 uM and didn't changed in concentration 300 uM. Activation of ATP-dependent K-channels by diazoxide did not affect the negative inotropic effect of NaHS. The obtained data suppose that in the mouse atrium exogenous and endogenous H2S causes a reduction of the force of contraction, which is mediated by the activation of ATP-dependent, calcium-activated or voltage-dependent K-channels.
Genes & Cells. 2014;9(3):94-98
pages 94-98 views

Development and applications of a new technique to study blood clot contraction (retraction)

Lozhkin A.P., Peshkova A.D., Ataullakhanov F.I., Litvinov R.I.


Despite the importance for hemostasis and thrombosis, platelet-governed clot shrinkage has not been systematically studied, partially due to the lack of methodology to follow and quantify clot contraction dynamics. We have developed a new technique based on the continuous tracking of clot size. An optical platform for this method is the commercially available Thrombodynamics Analyser System (HemaCore, Russia). A standard procedure includes recalcification of whole citrated blood and addition of thrombin to initiate blood clotting and platelet activation. The clot contraction is monitored by taking images every 15 seconds over 20 minutes or more followed by off-line computational analysis that provides a kinetic curve characterized by 6 numerical parameters. Clot contraction has at least two phases characterized by distinct rates. Exogenous Ca2+ is not indispensable for clot contraction to occur; however, the clots formed without addition of Ca2+ were less stable. Recalcification of blood with 2-5 mM [Ca2+] prevented the red blood cell fallout without an effect on the contraction kinetics. 10 mM [Ca2+] partially inhibited clot contraction. Iodoacetamide, an inhibitor of factor XIIIa, did not affect clot formation but abolished clot shrinkage, confirming that factor XIIIa is essential for clot contraction. Thrombin enhanced the rate and degree of clot contraction in a dose-dependent manner. Blood clot contraction was substantially delayed in patients on warfarin compared to healthy donors. We developed an accurate and simple assay for blood clot contraction which can be used for research and may be potentially useful for in vitro diagnostics.
Genes & Cells. 2014;9(3):99-104
pages 99-104 views

Pathohistological assessment of skeletal muscle after direct gene therapy with vegf165 of patients with peripheral arterial diseases

Mavlikeev M.O., Plotnikov M.V., Maksimov A.V., Gafiyatullina G.R., Murtazin A.I., Teregulov U.E., Shamsutdinova I.I., Gumerova A.A., Rizvanov A.A., Kiassov A.P.


The aim was to elucidate impact of gene therapy with plasmid encoding vegf165 on the muscle tissue pathohistology of patients with peripheral arterial diseases. Twice repeated intramuscular injections of plasmid («Neovaskulgen», RN LP-000671 from 28.09.2011) were performed to 6 patients (ischemia grade IIb-III by Pokrovsky-Fontaine) according to specification. Standart tredmill test, ankle-brachial index estimation were performed. Histological study of injured muscle biopsies taken before and 3 months after injection was performed. In intact muscles therapy with vegf165 leads to increase of mean cross-sectional muscle fiber area without significant angiogenic effect. In muscles with decreased capillary density this therapy leads to blood supply improvement promoting regeneration of muscles by myosatellites proliferation and increase of mean cross-sectional muscle fiber area and connective tissue degradation. Treadmill test showed painless walking distance increased by 31,74% on average (from 94,96±49,79 m to 139,11±60,78 m, p<0,05) in all patients. There was correlation of pathohistological analysis with clinical data.
Genes & Cells. 2014;9(3):105-111
pages 105-111 views

Deriving population of ectomesenchymal cells with properties of stem cells and progenitor cells from pulp of permanent teeth

Sagitov I.I., Shafigullina A.K., Saleeva G.T., Gomzikova M.O., Rizvanov A.A., Kiassov A.P.


Lately a lot of research has been focused on isolation of tooth stem cells. Our earlier studies showed that odontoblasts have a number of features typical for ectodermal cells, such as expression of epithelial membrane antigen (EMA) and cytokeratin 18. Aim: to obtain populations of ectomesenchymal cells from permanent teeth pulp with properties of stem and progenitor cells and characterize their phenotype in vitro. The pulp of permanent teeth of 15 patients (age 18- 29) was removed according to the orthodontic indications and used for stem cells isolation. The phenotype of derived cells was studied using the immunocytochemical staining method. Antibody staining for EMA, C-kit, cytokeratine 19, ESA, aSMA, Desmin, Vimentin, PCNA, Bcl-2. EMA and cytoceratine 18 expression was also confirmed by western blotting. The immunophenotype of obtained stem cells was determined by flow cytometry Significant expression of C-kit and Vimentin was observed at all times of cultivation as demonstrated by immunocytochemical staining. About a half of cells expressed EMA on all passages. First and second passages cells were characterized by expression of cytoceratine 18 which gradually decreased and to the fifth passage only single cells were positively stained. EMA and cytokeratin 18 expression was confirmed by western blotting. Flow cytometry revealed that cells obtained by us from human permanent teeth pulp were positive for CD29, CD90, CD10, CD54, CD56, CD166 and negative for CD14, CD34, CD45.
Genes & Cells. 2014;9(3):112-117
pages 112-117 views

Effects of platelet factor 4 on morphological and biochemical signs of apoptosis in T-lymphocytes

Skibo Y.V., Fathullina A.R., Romanova E.V., Litvinov R.I.


Platelet factor 4 (PF4) pertains to a family of CXC chemokines released by activated platelets. PF4 has a broad spectrum of effects on different cell types, including modulation of the immune response. In this study, we explore effects of PF4 on the morphological and biochemical signs of apoptosis in human T-lymphocytes in vitro. T-lymphocytes were isolated from peripheral blood of healthy donors using negative immunomagnetic separation and cultured in the complete RPMI 1640 medium for 24 hours in the absence and presence of PF4 added at a final concentration of 2 /ug/ml or 100 ug/ml. After 2, 4, 6, 12 and 24 hours of incubation the cells were studied with transmission electron microscopy and Western blot analysis with respect to potential apoptotic changes. The electron microscopy of control T-lymphocytes showed that the vast majority of the cells had a morphology characteristic of apoptosis at different stages. Adding PF4 at a concentration of 2 ug/ml reduced the number of cells at the late stages of apoptosis, while maintaining the signs of the early apoptosis in most of the T-lymphocytes. In the presence of 100 ug/ml PF4 nearly all of the cells kept a typical morphology of normal T-cells throughout the time of cultivation. The morphological apoptotic changes correlated well with expression of caspase-3, which was clearly detected in the control cells and cells treated with 2 ug/ ml PF4, but was almost abolished in the cells treated with 100 ug/ml PF4. Our results provide direct evidence for the dose-dependent anti-apoptotic effects of PF4 on T-cells, suggesting that PF4 sustains an immune response by extending T-lymphocyte survival.
Genes & Cells. 2014;9(3):118-124
pages 118-124 views

Application of recombinant histone protein H1.3 for inhibition of adenoviral infection

Solovyeva V.V., Rizvanov A.A.


There is an ongoing search for new drugs for the treatment of adenovirus infections. In our work the antiviral properties of the recombinant histone H1.3 in in vitro cell culture was investigated and for the first time we demonstrated that recombinant histone H1.3 significantly reduces the efficiency of adenoviral transduction. Also, recombinant histone H1.3 exerts an inhibitory effect on plaque formation on HEK-293A cells monolayer, infected with adenovirus serotype 5, which confirms antiviral properties of histone H1.3 towards adenoviruses.
Genes & Cells. 2014;9(3):125-130
pages 125-130 views

Hepatic stellate cells stimulate liver regeneration after partial hepatectomy under inhibition of hepatocyte proliferation

Titova A.A., Burganova G.R., Sharipova E.I., Pevnev G.O., Mavlikeev M.O., Gazizov I.M., Galyavieva A.R., Shafigullina A.K., Kaligin M.S., Titova M.A., Gumerova A.A., Kiassov A.P.


Most of the fundamental studies in the field of developing new methods for treating liver diseases in regenerative medicine are performed with haematopoietic and mesenchymal stem cells. At the same time more and more importance is gaining need for finding new approaches enabling stimulation of regional stem cell compartment and using regional stem cells which are possibly represented by hepatic stellate cells. But studies aimed at their transplantation are rare. Previously, we have established the possibility of these cells to differentiate into hepatocytes after transplantation to rats with partial hepatectomy. In our present research we studied liver regeneration after transplantation of hepatic stellate cells on partial hepatectomy with 2-acetylaminofluorene damage model in rats. 2-acetylaminofluorene blocks hepatocyte proliferation. Results of study confirmed that hepatic stellate cells have the ability to differentiate into hepatocyte and stimulate it's regeneration without the threat of fibrosis after transplantation in rats with partial hepatectomy and administration of 2-acetylaminofluorene.
Genes & Cells. 2014;9(3):131-135
pages 131-135 views

Experience of preserved corneal transplantation for penetrating keratoplasty

Taldaev R.E., Andreeva D.I., Kiassov A.P.


Corneal diseases are one of the leading causes of blindness and poor vision. Keratoplasty is the only effective method of treatment in case of corneal lesion or perforation. Since the use of the native cornea is restricted by the lifespan of the material and its limited availability, it is reasonable to use the preserved corneal transplants, especially in case of emergency. The analysis of 26 penetrating keratoplasties is adduced. One group of patients (22 eyes) were performed elective surgery, and the second group (4 eyes) - urgent intervention. Patients were follow-upped for 24 months. All patients were examined by standard methods, additionally; the amount of endothelial cell was counted. Transparent engraftment was achieved on 18 eyes (81%) in the first group and three eyes (75%) in the second group. Endothelial cell density was 1500±200/mm2. Visual acuity after surgery varies depending on the disease that caused keratoplasty, postoperative course, and the condition of posterior segment of the eye. Preserved cornea is a safe against blood transmitted infections, makes a penetrating keratoplasty possible in various diseases of the cornea, including the need for emergency surgery, and allows achieving a transparent and translucent engraftment in most cases.
Genes & Cells. 2014;9(3):136-139
pages 136-139 views

Pharmacogenetic and bioengineering approaches to the treatment of glial tumors of the brain

Bryukhovetskiy I.S., Bryukhovetskiy A.S., Khotimchenko Y.S.


The article analyzes the existing ideas about the specific mechanisms of therapeutic resistance of glial tumors of the brain, systematized the main trends in modern chemotherapy glial tumors, an attempt to justify the new bioengineering approaches to the creation of personalized cell preparations for therapy of glial tumors based on molecular - biological characteristics of tumor stem cells. It is shown that the main tool of the therapeutic effects can be own stem cells of cancer patients and tumor stem cell proteome can be considered as the primary target cell therapy.
Genes & Cells. 2014;9(3):140-146
pages 140-146 views

Possibility of application primary cell culture of kidney newborn rabbits in vaccines production

Glagoleva I.S., Plotnikova E.M.


Possibility of application of primary cell cultures in production of vaccines for domestic animals has been presented in this paper. Also the comparative description of application of organ cultures, cell lines and strains in virology has been provided. We have analyzed the different methods of receiving primary cultures. Also we presented our own data of primary cell cultures sensitivity to various virus agents of domestic animals on the example of the cell culture of kidney newborn rabbits.
Genes & Cells. 2014;9(3):151-154
pages 151-154 views

Programable biopolymeric microand nanocontainers for biocomputers

Erokhina S., Pastorino L., Sorokin V., Erokhin V.


Polymeric capsules (containers) are widely used now in researches connected to the design of new pharmaceutical preparations. Here we present data allowing to consider these objects also as essential parts of biocomputers, capable, in particular, to carry out targeted delivery and triggered start of the «main program» (encapsulated specific molecules) to the areas with damaged functioning of the system, resulted from undesirable reactions. We also consider the use of cells as program package, capable to perform parallel functions in biocomputers when delivered in a proper place and started functioning in a proper moment.
Genes & Cells. 2014;9(3):155-159
pages 155-159 views

Cholinesterases: the opinion of neurophysiologist

Petrov K.A., Kharlamova A.D., Nikolsky E.E.


The review addresses issues of structure and functions of acetyl- and butyrylcholinesterases. Authors consider these enzymes not only as limiters of the neurotransmitter acetylcholine life span in synaptic cleft but also accounting for their putative non-synaptic functions. Particular emphasis has been placed on the possibility of correction of nerve system pathologies by way of modification of the activity of these enzymes.
Genes & Cells. 2014;9(3):160-167
pages 160-167 views

Cell technologies for muscle tissue restoration. Part II: skeletal and smooth muscles

Korsakov I.N., Zorin V.L., Eremin I.I., Zorina A.I., Kotenko K.V., Pulin A.A.


This review represents ongoing clinical studies devoted to recovery of muscle tissue using cell technologies, as well as analysis of applied populations of cells.
Genes & Cells. 2014;9(3):168-172
pages 168-172 views

Features of senescence associated secretory phenotype (sasp) in gastrointestinal stromal tumors (GIsTs)

Ramazanov B.R., Boichuk S.V., Rizvanov A.A.


To study the cellular senescence mechanisms and senescence-associated secretory phenotype of the tumor cell lines in response to doxorubicin exposure. The cell lines indicated above were used in present study: human fibroblasts lines BJ, human osteosarcoma cell line U-2 OS and gastrointestinal stromal tumor cell line GIST-T1. Genotoxic stress was induced by the transient exposure of doxorubicin (0.25 ug / ml) for 5 hours. DNA double-strand breaks formation was assessed by an increased expression and focal accumulation of the phosphorylated form of histone 2A (y-H2AX), whereas the activation of DNA repair pathway was assessed by focal accumulation of the phosphorylated form of ATM-kinase (pATM Ser1981). An increase of p-galactosidase activity was estimated by X-gal assay (5-bromo-4-chloro-3-indo-beta-D-galactoside). Interleukin-6 (IL-6) and interleukin-8 (IL-8) production was assessed by ELISA at 3, 6, 9, 12 days after 5h exposure to doxorubicin. Incubation of the tumor cell lines with doxorubicin induced DNA damage which triggered the DNA damage response (DDR) and activated a cellular senescence program associated with typical morphological changes, decrease of Lamin B1 expression and accumulation of p-galactosidase in the cytoplasm known as a traditional senescence markers. We observed a significant increase of IL-6 and IL-8 production in doxorubicin-treated human fibroblasts and tumor U-2 OS cells, whereas GIST-T1 cells secreted IL-6 only. In contrast to non-transformed cells, tumor cells after being exposed to doxorubicin triggered a permanent DNA damage, activation of cellular senescence program and production of proinflammatory cytokines (IL-6 and -8). Senescence-associated secretory phenotype (SASP) in GIST-T1 cells differs from the phenotype observed in the other of cancer cells and normal cells, as well. This might be taken into account during the development of the novel treatment options for the patients with gastrointestinal stromal tumors (GISTs).
Genes & Cells. 2014;9(3):173-178
pages 173-178 views

Genetic effects of medicines - lipid exchange modulators

Ibragimova M.Y., Semenov V.V., Kh Ibragimov Y.K., Validov S.Z., Zhdanov R.I.


There are medicines known as adrenoreceptors ligands, which influences a variety of lipid exchange, therefore the study of their mutagenic and antimutagenic potentials, their genotoxicity, is of great interest. The study are fullfiled using C57B4/6 mice (males) of 20 g and 1,5-2 months of age. Mutagenic and antimutagenic effects of the ligands were evaluated using in vivo tests: mouse brain skeleton cells (a number of chromosomal aberrations) and mouse peripheral red blood cells peripheral (a number of cells with micronuclei from 2000 analyzed cells). 6 male mice were used for every experimental and control variant. All medicines studied - epinephrine, phenylephrine, orciprenalin, proroxan, and propranolol (besides epinephrine, 50 mg/kg) - in a majority of doses used don't increase the level of chromosomal aberrations and the number of red blood cells with micronuclei.Epinephrin, phenylephrine, orciprenalin in doses studied demonstrated a protector effect and decreased a level of chromosomal aberrations and a number of red blood cells with micronuclei induced by cyclophosphamide administration. Proroxan and propranolol didn't change a level of aberrations. It was shown that adrenoreceptor ligands decreasing or increasing lipid level did not express mutagenic effect, and, in opposite, expressed antimutageni which testify that the system participating simultaneously in a realization of lipid exchange and mutagenesis/ antimutagenesis exists. An action of the system is realized via cell membrane receptors. According mechanism of action, the system is polyreceptoric (a- and р-adrenoreceptors take a part) and polyvalent, namely, receptor stimulators and blocators take a part in a formation of cell antimutagenesis.
Genes & Cells. 2014;9(3):199-203
pages 199-203 views

Genetically encoded calcium indicator GCaMP6m for two-photon calcium imaging in newborn mouse cerebral cortex

Akhmetshina D., Villette V., Tressard T., Malvache A., Khazipov R., Cossart R.


Calcium is a universal messenger regulating essential cellular signaling events in many tissues and organisms. In neurons, action potentials trigger large and rapid changes in cytoplasmic-free calcium concentration. Genetically encoded calcium indicators (GECIs) have been iteratively improved and are becoming useful for quantitative imaging of activity in defined neuronal populations in vivo. Among them, GCaMP6m has been recently introduced as an ultrasensitive calcium sensor for the experiments in vivo. Whether GECIs can be used in developmental studies to characterize early activity patterns in the immature cerebral cortex remains unknown, however. We used in vivo two-photon calcium imaging with GCaMP6m calcium indicator to characterize its developmental expression following adeno-associated virus (AAV) mediated viral induction of GCaMP6m expression in newborn mice (at postnatal days P0-2). Three days after virus injection, the vast majority of neurons showed fluorescent labeling in the neuronal cytoplasm, dendrites and axons. We found that the GCaMP6m calcium sensor is expressed in the cerebral cortex within 3-5 days following transfection at levels that enable monitoring spontaneous and sensory-evoked calcium transients in the intact mice barrel cortex in vivo. Thus, following GCaMP6m transfection at birth, it is possible to monitor neuronal populations in the barrel field of somatosensory cortex in P5-7 mice in vivo.
Genes & Cells. 2014;9(3):179-184
pages 179-184 views

On the problem of lipid coding of genomic DNA: molecular dynamics peculiarities of DNA-phospholipid complexes

Zhdanov R.I., Aupov R.K., Andrianov G.V., Akberova N.I., Ibragimova M.Y.


Lipids can be bound to nucleic acids forming coding sequence at genomic DNA, and participating in realization of language of genome. Interaction of DNA oligonucleotide(A)20-(T)20 is studied with by moleculat dynamics method. The complex is prepared as a result of molecular docking, phospholipid molecule being located at DNA minor groove. Binding energy is equal to - 6.3 kcal/mol. Formation of hydrogen binding, interactions leading to those bonds, and some other interactions between DNA and phospholipid. The number of those interactions due to molecular dynamics is varied between 175 and 337. The key role in stabilization of DNA-phospholipid complex play also van-der-vaals and hydrophobic interactions along with hydrogen binding. The role of ethanolamine grouping in complexation between DNA and phospholipid is demonstrated.
Genes & Cells. 2014;9(3):185-190
pages 185-190 views

Complement system d-inhibitor at newborns of intrauterine infection development risk group

Zhdanova S.I., Bikmullina Z.M., Kozlov L.V., Bulatov V.P., Gora N.V., Andina S.S., Zhdanov R.I.


Complement system is considered as unspecific defence system of human organism, which starts to function just after born. The state of complement system at newborns including C1 inhibitor data is less studied. The paper is aimed to study functional activity and contents of C1 inhibitor in blood of newborn infants (taken on 3-5 day of life), their mothers and umbilical blood as well from group of the risk of development of mycoplasma and ureaplasma infections. Totally 105 blood samples taken from newborn infants and their mothers of risk group were analysed based on occasional selection approach. Ten healthy newborn infants without risk of intrautero infection development and their mothers were used as a control group. Complement C1 inhibitor functional activity and contents is studied by immune enzyme assay using ELISA. C1 inhibitor activity and blood contents at newborn infants from mothers of infection risk group is higher compare to newborn infants from healthy mothers. Infection agent induces C1 activity more than its content at mother blood, due to higher turnover of this protein. It is approved the increased C1 activity at newborn blood of control group (p = 0,009) and of mothers of risk group (р = 0,0009) compare to umbilical blood. The mothers' blood is characterized with very high level of C1 content and functional activity many times higher than normal values which is specific to the acute phase state of the organism during the born process. C1 content at umbilical cord blood is 4-fold less than at mothers blood which testifies an existence of blood-brain barrier for C1 inhibitor.
Genes & Cells. 2014;9(3):191-194
pages 191-194 views

Invasive ability of proteus mirabilis and morganella morganii

Zamaliutdinova N.M., Bogomolnaya L.B., Chastukhina I.B., Sharipova M.R., Mardanova A.M.


Here we discuss the ability of Proteus mirabilis 5127-1 and Morganella morganii ZM to invade human cervical epithelial adenocarcinoma cells (HeLa-M) and to cause cytotoxicity in those cells. We found that ability of bacteria to invade HeLa-M cells depended on the multiplicity of infection for both strains. Cytotoxicity of P. mirabilis on HeLa-M cells was evaluated through quantification of eukaryotic cell detachment from plastic. We hypothesized that cytotoxicity of P.mirabilis could be due its extracellular proteolytic and hemolytic activities. In contrast, no cytotoxicity was detected in HeLa-M cells infected with M.morganii.
Genes & Cells. 2014;9(3):195-198
pages 195-198 views

Controllable endogenic proteolysis as a major factor of switching tryptophanyl-tRNA synthetase from canonical aminoacylation activity to non-canonical regulatory functions

Nurbekov M.K., Elov A.A., Zhdanov R.I.


Tryptophanyl-tRNA synthetase has a unique set of additional non-canonical activities in addition main aminoacylation activity, in particular for the control of angiogenesis [7]. It is important to study the role of endogenous limited controlled proteolysis as a possible molecular mechanism of switching from the canonical source aminoacylation activity of the native enzyme to non-canonical anti-angiogenic activity. We consider important data on the possible role of the significant for the activity zinc ion discovered earlier in the tryptophanyl-tRNA synthetase [8], and endogenous tryptophan in the activation of the regulatory functions of the enzyme. We have developed a novel approach to the identification of the role of endogenous proteolysis induced by intracellular proteases in the modification of the enzyme, as well as the role of the endogenous enzyme bound tryptophan and varying content of zinc ion essential for tryptophanyl-tRNA synthetase activity, as important factors in the complex process of activation of the non-canonical cytokine functions of the enzyme. Proteolytic transition of native tryptophanyl-tRNA synthetase as a result of above modifications of the enzyme and resulting cleavage of N-terminal fragment of 20 kDa have been shown with chromatographic, immunochemical methods, and confirmed by electrophoresis and immunoblotting.
Genes & Cells. 2014;9(3):223-229
pages 223-229 views

Investigation of recombinant therapeutic genes expression in umbilical cord blood mononuclear cells transduced with three adenoviral vectors encoding neurotrophic factors GDNF, VEGF and neural cell adhesion molecule NCAM

Islamov R.R., Rizvanov A.A., Cherenkova E.E., Mukhamedshina Y.O., Salafutdinov I.I., Safiullov Z.Z., Izmailov A.A., Mukhamedyarov M.A., Guseva D.S.


To overcome the effects of neurodegeneration and stimulating neuroregeneration in the spinal cord transgenic mice with model of amyotrophic lateral sclerosis were injected intraspinally with umbilical cord blood mononuclear cells (UCBMCs) transduced with adenoviral vectors encoding for vascular endothelial growth factor, glia-derived neurotrophic factor and neural adhesion molecule. Two weeks after transplantation UCBMCs were localized in the site of injection and at a distance from it, confirming the migration capacity of the cells in spinal cord parenchyma. Using triple immunofluorescence staining UCBMCs expressing therapeutic genes were found. The study results suggest the expedience of given gene-cell construct to stimulate the neuroregeneration in central nervous system after neurotrauma, ischemic stroke and in neurodegenerative diseases.
Genes & Cells. 2014;9(3):204-208
pages 204-208 views

Binase induces pro-inflammatory immune response of macrophages

Lozhkin A.P., Zelenikhin P.V., Makeeva A.V., Sokolova E.A., Ilinskaya O.N.


Biopharmaceutical agents, especially proteins, may possess a broad spectrum of immunological effects. Cytotoxic ribonucleases, such as binase - RNase of Bacillus pumilus, have a significant potential as anticancer therapeutical agents. However without comprehensive analysis of immune system reactions on the proteins, the practical utility of them is complicated. Using the flow cytometry we estimated qualitative and quantitative changes in IL-6, IL-10, МСР-1 and TNF-acytokines synthesis by peripheral macrophages of mice under the influence of binase and RNAse A. We established that binase in concentration 100 and 300 jg/ml, in contrast to RNase A, is able to activate pro-inflammatory response of macrophages, inducing increase of IL-6, МСР-1 andTNF-a cytokines level in cultural media. Treatment with RNases does not enhance the antiinflammatory cytokime IL-10 synthesis. Obtained data lead as to the assumption, that binase capability to activate pro-inflammatory response of macrophages will intensify its antitumor effects in vivo.
Genes & Cells. 2014;9(3):209-211
pages 209-211 views

Phenotypic modifications and quantitative analysis of glial cells in the area of spinal cord injury at the cell-mediated and direct gene delivery gdnf

Mukhamedshina Y.O., Shaymardanova G.F., Mukhitov A.R., Garanina E.E., Rizvanov A.A., Chelyshev Y.A.


On the model of rat spinal cord dosed contusion at T8 level studied the effect of delivery into the area of damage of the cell-mediated and direct gene delivery GDNF on maintaining a population of glial cells. Delivery into the area of damage adenoviral vector with gene GDNF (AdV-gdnf) using human umbilical cord blood mononuclear cells has a greater influence the amount of Ch47+-cells in the ventral horn (VH). Direct delivery of AdV-gdnf influences the amount of Ch47+-cells in dorsal roots entry zone (DREZ). Cell-mediated gene delivery GDNF causes the most pronounced changes in the expression of marker proteins astrocyte GFAP, S100B, and AQP4 in the ventral funiculus (VF) of white matter. Cell-mediated and direct delivery of gene GDNF support population GFAP+/S100B+-cells. The results indicate that the direct and cell-mediated gene delivery GDNF into spinal cord injury have different effects on the populations of glial cells in specific areas of spinal cord, that is important for the optimal method of delivery of therapeutic genes to stimulate posttraumatic neuroregeneration.
Genes & Cells. 2014;9(3):217-222
pages 217-222 views


Mijanovic O., Zhuravleva M.N., Gomzikova M.O., Rizvanov A.A., Kiassov A.P.


In recent years there were lots of researches focused on inhibiting connective tissue cells in the liver of patients with chronic hepatitis. Most studies of the substances which can potentially be considered as drugs for the treatment of liver fibrosis were performed on hepatic stellate cells. However, there is a theory of portal fibroblasts participation in liver fibrosis, as in most cases the processes of fibrogenesis start both at main portal tract and intrahepatic ducts. Suppression of fibrogenesis by preventing the activation and induction of apoptosis of portal fibroblasts are being considered as therapeutic strategies for the treatment of liver fibrosis.
Genes & Cells. 2014;9(3):212-216
pages 212-216 views

Muscarinic regulation of calcium transient and synaptic transmission in frog neuromuscular junction

Samigullin D.V., Khaziev E.F., Kovyazina I.V., Bukharaeva E.A., Nikolsky E.E.


In frog neuromuscular junction, muscarine, exogeneous acetylcholine and acetylcholinesterase inhibitor proserine reduced the intensity of calcium-sensitive dye fluorescence (calcium transient) at low frequency of nerve stimulation, suggesting that calcium ions entry into nerve ending was decreased. M2 muscarinic receptor blocker methoctramine prevented the action of muscarine. The amplitude of endplate currents was reduced in presence of muscarine at low frequency nerve stimulation, and atropine abolished this effect. Amplitudes of endplate currents evoked by high frequency stimulation were enhanced in presence of methoctramine, and synaptic depression was less pronounced, probably due to elevated calcium concentration in nerve ending. Thus, activation of presynaptic muscarinic receptors predominantly of M2 subtype reduces the intensity of quantal acetylcholine release in frog neuromuscular synapses that may be associated with decreased level of calcium ions in the nerve ending to provide the modulation of postsynaptic currents amplitude at high frequency firing.
Genes & Cells. 2014;9(3):242-247
pages 242-247 views

The influence of P2-receptors on the cytotoxic effect of cisplatine in squamous cell uterine cervix cancer in vitro

Raginov I.S., Muhamedyarova G.K., Panov A.V.


To investigate the influence of purinergic signalling in squamous cell uterine cervix cancer in vitro, the effect of combining ATP (nonselective agonist P2-receptor) with the nonselective P2 receptor antagonists PPADS (P2X-receptors), RB2 (P2Y-receptors) and suramin (both P2Y-receptors and P2X-receptors) and the chemotherapeutic drug cisplatin was assessed. ATP effectively reduces up to twofold the growth of uterine cervix cancer cells. RB2 promoted survival of tumor cell by 52,1% compared to the control. Combinations of the cisplatin + ATP demonstrated a strong sensitivity at 2,7 fold. However, this effect of ATP was reversed in the presence of RB2 by 41%. This highlights the potential use of selective P2Y receptor agonists in the treatment of squamous cell uterine cervix cancer.
Genes & Cells. 2014;9(3):240-241
pages 240-241 views

Transplantation of mononuclear cells of human umbilical cord blood improves spatial memory in APP/PS1 transgenic mice with alzheimer's disease model

Petukhova E.O., Mukhamedshina Y.O., Rizvanov A.A., Mukhitov A.R., Zefirov A.L., Islamov R.R., Mukhamedyarov M.A.


Alzheimer's disease is progressive incurable neurodegenerative disease, which is manifested mainly by dementia. One of the most promising directions in development of Alzheimer's disease treatment is use of gene-cell technologies. The aim of current study was to perform transplantation of wild-type or EGFP expressing umbilical cord blood mononuclear cells (МКПК) to APP/PS1 transgenic mice with Alzheimer's disease model with further evaluation of transplantation impact with behavioral (T-maze, plus maze, open field) and immunohistochemical methods. It was found that МКПК transplantation significantly ameliorates behavioral performance of APP/PS1 mice: improves spatial memory, decreases anxiety and non-specific excitability, increases the efficacy of exploratory behavior. Grafted cells were found in cortex and hippocampus of mice even 3 months after МКПК transplantation, herewith EGFP expression in grafted cells was found at early stages after transplantation. Thus, use of МКПК-based gene-cell constructs represents a promising direction in development of Alzheimer's disease therapy.
Genes & Cells. 2014;9(3):234-239
pages 234-239 views

Otsenka sokratitel'noy funktsii diafragmal'noy myshtsy mSOD1 transgennykh myshey s model'yu bokovogo amiotroficheskogo skleroza

Nurieva L.R., Islamov R.R., Grigor'ev P.N., Rizvanov A.A., Zefirov A.L., Mukhamed'yarov M.A.


Amyotrophic lateral sclerosis (ALS) - incurable neurodegenerative disease, which is characterized by progressive loss of motoneurons, atrophy and paralysis of skeletal muscles. The main immediate cause of death in ALS patients is a paralysis of the respiratory muscles. In the present study we investigated the contractile characteristics of diaphragm muscle of mSOD1 transgenic mice with ALS model at presymptomatic (3-4 MO) and symptomatic (6-7 MO) stages of the disease. In myographic experiments under electrical stimulation of muscle we showed an increase in force and duration of a single muscle contraction, force of summed muscle contractions (5-50 Hz), diaphragmatic muscle mass at mSOD1 symptomatic mice compared to presymptomatic and wild-type mice. The absence of changes in the contraction threshold and «force-stimulus» curve at symptomatic mSOD1 mice suggests that found differences are not related to changes in the excitability of muscle fibers. Results of the study may be explained in terms of activation of compensatory mechanisms in the «motoneuron - neuromuscular synapse - skeletal muscle» in response to the development of neurodegenerative process. Presented findings extend current understanding of the pathogenesis of amyotrophic lateral sclerosis and can be used to estimate the rate of progression of the disease in mSOD transgenic animals.
Genes & Cells. 2014;9(3):230-233
pages 230-233 views

The action of bacillary proteases on matrix metalloproteinases of different cell lines

Toymentseva A.A., Danilova Y.V., Balaban N.P., Mardanova A.M., Sharipova M.R.


It was investigated the effect of secreted proteinases Bacillus pumilus 7P belonging to different classes of proteolytic proteins on matrix metalloproteinases of mouse fibroblast (3T3BalbSV40), multipotent mesenchymal stromal and cells Hela-M. Matrix metalloproteinases of all cell lines was inactivated by glutamyl-specific endopeptidase. Subtilisin-like proteinase and adams-like metalloproteinase of B. pumilus did not change the activity of matrix metalloproteinases of differentcell lines.
Genes & Cells. 2014;9(3):253-257
pages 253-257 views

H. pylori, mikrobiota kishechnika, antibiotikorezistentnost': est' li vzaimosvyaz'?

Safina D.D., Abdulkhakov S.R., Abdulkhakov R.A., Ismagilova R.K., Tyakht A.V., Popenko A.S.


Helicobacter pylori (H. pylori) eradication is obligatory in case of such diseases as gastric and duodenal ulcer, atrophic gastritis, MALT lymphoma, in patients after gastrectomy due to the gastric cancer, as well as in close relatives of gastric canсer patients. Antibacterial therapy which is aimed at the eradication of H. pylori has an impact on the intestinal microbiotа leading to its qualitative and quantitative disturbances. The review focuses on the influence of H. pylori and eradication therapy on the content of the intestinal microbiota, their role in the development of intestinal microbiota resistance to antibiotics. Both H. pylori itself and eradication therapy can lead to the changes of gut microbiota content, which are characterized by reduction in microorganisms diversity; decrease in the number of obligate anaerobes, prevailing in normal gut; increase in the number of facultative aerobic microorganisms. Long-term effect of antibiotic therapy is the development of antibiotic-resistant strains of bacteria. Resistance may be due to the presence of the resistance genes in bacteria, which they have acquired through mutations or horizontal transfer. These genes can be spread not only among pathogenic bacteria, due to the presence of which antibacterial therapy is carried out, but also among the members of the normal microflora, which can then be modified and loose its beneficial properties. Thus the intestinal microflora may serve as a potential reservoir of resistance genes and contribute to the development of antibiotics resistance through other members of the human microbiota.
Genes & Cells. 2014;9(3):248-252
pages 248-252 views

Rapid Diagnostic Test of Rabies Using Rat Gasser's Ganglion Neurinoma Cell Culture (RGGN-1)

Khismatullina N.A., Gulyukin A.M., Shuralev E.A., Khaertynov K.S., Chernov A.N., Filimonova M.N., Avzalova A.F., Parshikova A.V., Ivanov A.V.


The results of the comparative study of the diagnostic efficacy of the method of isolation of street rabies virus using rat Gasser‘s ganglion neurinoma cell culture (RGGN-1) and biological tests on white mice are presented. It is shown that the method of virus isolation using RGGN-1 cell culture is not inferior in the sensitivity to the classic bioassay on white mice and has obvious advantages, such as reducing the time of diagnosis (up to 3 days using RGGN-1 compared to 30 days using white mice) and its cost-effectiveness. Method of cell line RGGN-1infection is recommended for inclusion in the Standards for the rapid diagnosis of rabies and isolation street rabies virus, which should be considered as a possible replacement of bioassays on white mice. Rapid diagnosis of rabies using RGGN-1 cell culture will provide early diagnosis of rabies in animals to reduce the risk of disease in human and animals.
Genes & Cells. 2014;9(3):276-280
pages 276-280 views

Helicobacter pylori infection in children in Kazan and factors influencing the effectiveness of eradication therapy

Khalikova A.R., Faizullina R.A., Akhmetov I.I., Ivanovskaya K.A., Abdulkhakov S.R.


The effectiveness of H. ру/ог/ eradication therapy depends on many factors, including the patients' compliance, H. ру/ог/ sensitivity to antibiotics, the rate of proton pump inhibitors metabolism in the liver. The Aim of this study was to investigate the frequency of mutations underlying the development of H. ру/ог/ resistance to clarithromycin and CYP2C19 genetic polymorphism in children with gastroduodenal pathology. The study included 91 patients with different types of gastroduodenal pathology. H. ру/ог/ was revealed using urease breath test (Helic-test, Russia), polymerase chain reaction (PCR), and cytology. In 26 children in whom the presence of H. ру/ог/ was confirmed by PCR, the presence of mutations (for three sites - A2142G, A2143G, T2717C) in the 23S RNA causing H. ру/ог/ resistance to clarithromycin was analysed. In 80 patients cytochrome P450 2C19 polymorphism was studied as well. H. ру/ог/ was detected by urease breath test in 80 children (87.9%); by PCR - in 54 out of 91 (59.3%). A2143G mutation in the 23S RNA gene was found in 2 out of 26 children (7.7%). A2142G and T2717C mutations were not detected in any case. It was shown that 61 patients out of 80 (76.3%) were CYP2C19*1/*1 genotype carriers; 19 (23.8%) had CYP2C19*1/*2 or CYP2C19*1/*3 genotype. CYP2C19*2/*2, CYP2C19*3/*3 and CYP2C19*2/*3 genotypes weren't found in any patients. The level of H. ру/ог/ clarithromycin resistance in children is 7.7%. The phenotypic group referring to the rapid proton pump inhibitors metabolizers is predominant (76.3%) in children with H. ру/ori-associated gastroduodenal pathology.
Genes & Cells. 2014;9(3):272-275
pages 272-275 views

Activity index of lymphocyte apoptosis in children with neonatal sepsis

Khaertynov K.S., Boichuk S.V., Anokhin V.A., Ramazanov B.R., Dunaev P.D., Khaiboullina S.F., Rizvanov A.A., Andreeva A.A., Satrutdinov M.A.


We studied the lymphocyte apoptosis in 15 infants with late neonatal sepsis, whereas 8 of them (53.3%) were considered as a preterm infants. Etiology of sepsis was identified in 4 cases (26.7%): in 1 case of each, the cause of the disease was Klebsiella pneumoniae, Pseudomonas koreenses, Candida and St. agalactae. Control group consisted of 5 healthy newborns. Apoptosis analysis was conducted by measuring the amount of hypodiploid cells by using a propidium iodide (Sigma Aldrich) DNA staining procedure and flow cytometry (FACsCanto II, Becton Dickinson). We observed an elevated numbers of apoptotic cells in all cases of neonatal sepsis. The most significant difference was observed when the lymphocytes were cultured for 3 and 5 days. For example, on day 3 of culture the numbers of apoptotic cells in patients with neonatal sepsis vs control group were 19.6% and 5.13%, respectively. Absolute lymphopenia was noted in 26.7% of cases with neonatal sepsis. No difference in the lymphocyte apoptosis between term and preterm infants was observed. An increased amount of lymphocyte apoptosis was not associated with C-reactive protein level during the whole time-points of the experiment: at 1, 3 and 5 days. Acute phase of neonatal sepsis is associated with an increased apoptosis of peripheral blood lymphocytes.
Genes & Cells. 2014;9(3):267-271
pages 267-271 views

Immunohistochemical study of motoneurons of the lumbar spinal cord of mice after the 30-days space flight on biosatellite BION-M1

Tyapkina O.V., Rezvyakov P.N., Nurullin L.F., Petrov K.A., Nikolskiy E.E., Islamov R.R.


We made immunohistochemical study of the lumbar spinal cord of c57black/6 mice after a 30-days space flight using antibodies against proteins which involved in providing of synaptic transmission (synaptophysin and PSD95). Also we use antibodies against proteins of intracellular protection (Hsp25 and Hsp70), and neuroprotective protein (VEGF) and its receptor (Flt-1). We found, that the levels of synaptophysin immune expression decreased by 21%, PSD - by 55%, Hsp25 - by 15% and Hsp70 - by 9% in mice lumbar spinal cord motoneurons after space flight. Thus, there were no significant changes in the immune expression of VEGF and its receptor Flt-1. Obtained data reveal the changes in the functional state (synaptic activity and resistance to stress) of mice lumbar spinal cord motoneurons after space flight. So, we have shown a new data about spinal cord motoneurons role in the development of hypogravity motor syndrome.
Genes & Cells. 2014;9(3):263-266
pages 263-266 views

The combination of intestinal chelators for correction of environmentally caused by trace element imbalance in children at the cellular level

Tunakova J.A., Fayzullin R.I., Valiev V.S.


Normally, the human body maintains a certain balance of micronutrients. The danger of a constant inflow of toxic elements from the environment caused by the inability to selfdestruction and elimination from the body, so the search for an optimal balance correction of metals is carried out. By AAS definition of the nine most common toxic and essential trace elements in hair of children in all 11 children's clinics service areas was held: Pb, Cu, Zn, Cr, Sr, Mn, Ni, Fe, Cd. Six different types of biopolymer ehnterosorbents: pectin, alginates, chitosan, microcrystalline cellulose and chitin were studied in the proposed model. A method of produce and combination of biopolymer and mineral enterosorbents with qualitatively new properties and lower cost compared to monosorbents was developed. 11 most common toxic and essential trace elements in hair of children were analyzed. The most variable and requiring of geographically differentiated correction elements in areas of research were determined. The most effective biopolymer enterosorbents and their combinations with mineral enterosorbents were discovered. Experimentally a method of producing composite enterosorbents based colloids was created. The highest sorption capacity related to of Zn, Cd, Fe and Cu requiring geographically differentiated correction was noted in combination of octahedral smectite and chitosan tests.
Genes & Cells. 2014;9(3):258-262
pages 258-262 views

Rapid flow cytofluorometric analysis of functional mitochondrial status in Yarrowia lipolytica yeast: a reasonable model to study human cell bioenergetic responses

Cherepnev G.V., Igtisamova G.R., Zainullin L.I., Kalacheva N.V., Rizvanov A.A.


We developed a rapid method to study Yarrowia lipolytica single yeast cell mitochondrial responses based on flow cytometry and potential-sensitive ratiometric fluorochrome JC-1. The proposed model has several advantages as it allows to measure mitochondrial membrane potential in intact cells (without isolation of mitochondria), has an internal positive control (uncoupler of oxidative phosphorylation CCCP), and produces a normalized measurement of the transmembrane mitochondrial potential, taking into account the heterogeneity of cell size and number of mitochondria in a single cell. The similarity of the mitochondrial complex I structure in mammals and Yarrowia lipolytica opens prospects for the model implementation in search for human bioenergetic response modifiers.
Genes & Cells. 2014;9(3):281-283
pages 281-283 views

The spread of mobile genetic elements SCCmec-type strains Staphylococcus Aureus, isolated with skin and mucous of patients with allergic pathology

Shamsutdinov A.F., Toymentseva A.A., Tyurin Y.A., Bayazitova L.T.


Staphylococcus aureus is widespread bacterium that causes inflectional diseases of humans and animals. Currently, there is growing incidence of methicillin-resistant strains of Staphylococcus aureus. Resistance of Staphylococcus aureus strains to methicillin is detected by the presence in the genome of staphylococcal chromosomal cassette of SCCmec type. In this work 60 clinical strains of MRSA isolated from the skin and mucous of patients with allergic pathology were examined. The analysis of Staphylococcus aureus strains was performed in differential mediums. Additionally, specific identification was carried out by MALDI-TOF. Specific regions of the genes of mec gene complex were identified by PCR. As a result of typing we determined that 47% of tested strains were resistant to oxacillin phenotypically, but did not contain the specific primer sites related to mec- gene complex types I-IV. The remaining 53% of the strains were contained primer-specific amplicons. These strains were also phenotypically resistant to oxacillin. This study showed that MRSA strains as part of its genetic apparatus are several different types of SCCmec cassettes. This property allows them to express multidrug resistance.
Genes & Cells. 2014;9(3):284-288
pages 284-288 views

Intracellular localization analysis of a conjugate of recombinant histone H1.3 with photoactivated fluorescent dye

Shaposhnikov M.N., Solovyeva D.O., Zaitsev S.Y., Solovyeva V.V., Salafutdinov I.I., Rizvanon A.A.


Histone proteins are promising carriers for delivery of recombinant nucleic acids in various cell cultures. Conjugates of proteins with fluorescent dyes are modern tools in the study of cellular transport and intracellular distribution of important biopolymers. The aim of this work was to optimize the method of recombinant histone H1.3 conjugation with photoactivatable fluorescent dye. Also we studied intracellular penetration and subcellular localization of the resulting conjugate. Resulting conjugate was capable of intensive fluorescence in the red region of the spectrum after irradiation with violet light. The degree of conjugation was determined under different reaction conditions. A comparison of the cytotoxicity of histone H1.3 and its conjugate with photoactivated fluorescent dye in HeLa cells and its intracellular localization was characterized. We found that conjugates mainly localize in recycling endosomes and to a lesser extent in peroxisomes. Thus, the resulting conjugate of recombinant histone H1.3 with photoactivatable fluorescent dye can be used for further studies its anticancer activity and as carrier for drugs and nucleic acids delivery into human and animal cells.
Genes & Cells. 2014;9(3):289-294
pages 289-294 views

Cytotoxic activity of plant oils of Nigella sativa, salvia officinalis and peptide metabolites of streptomyces mirabilis fK749 against HeLa and MCF-7 cancer cell lines

El-Shafei S.M., Abd El-Rahman A.A., Validov S.Z., Bikmullin A.G., Sanatova E.R., Zalyalyutdinova L.M., Fattakhova A.N., Alimova F.K.


In the last several years, there has been an increase in the use of plant materials in clinical development of new drugs and methods in the treatment of neoplastic diseases. In this regard, the current study was aimed to investigate the cytotoxic activity of Nigella sativa (NS), Salvia officinalis (SO) plant oils and peptide metabolites (PM) of Streptomyces mirabilis FK749 on the viability of Hela and MCF-7 cancer cells. To achieve this objective, cells were exposed to 0, 20, 40, 60, 80 and 100 mg/ml of NS, SO plant oils and PM S. mirabilis FK749. After 24 h. incubation the effect of plant oils and PM on the viability and apoptotic activity of the cancer cells were assessed by using trypan blue and TMRE assays. Morphological changes were examined. Whereas, the half-maximal inhibitory concentrations of these oils and PM were assessed after 72 h. incubation. The results of this study showed that treatment Hela and MCF-7 cells with NS, SO and PM at 40 mg/ml and above concentrations up to 100 mg/ml were found to be cytotoxic and leads to a decrease in the cell viability. The morphological changes observed in Hela and MCF-7 cells exposed to different concentrations of NS SO and PM were found to be in a concentration dependent manner. Hela cells were shown to be more resistant against oil components and PM than MCF-7 cells.
Genes & Cells. 2014;9(3):295-302
pages 295-302 views

Application of stromal vascular fraction cells of adipose tissue for treatment of hyaline cartilage defect of the medial femoral knee condyle: case report

Masgutov R.F., Salihov R.Z., Rizvanov A.A., Salafutdinov I.I., Bogov A.A., Plaseichuk Y.A.


The article presents a case report of successful treatment of the 50 years old female patient with osteoarthritis of the knee with autologous cells stromal vascular fraction (SVF) from adipose tissue. SVF autologous cells were transplanted into the local cartilage defect of the medial femoral condyle composed of fibrin glue. In 5 months after the operation the patient noted pronounced decrease the pain, increase range of motion in the knee joint. According to the MRI the almost complete decrease of subchondral edema, absence of synovitis were observed. The follow-up was 3 years.
Genes & Cells. 2014;9(3):303-306
pages 303-306 views

Recovery of ulnar nerve defect by autonerve grafting and autotransplantation of stromal vascular fraction of adipose tissue

Masgutov R.F., Gallyamov A.R., Shulman A.A., Bogov (jr) A.A., Khannanova I.G., Mullin R.I., Salafutdinov I.I., Topyrkin V.G., Rizvanov A.A., Bogov A.A.


This article describes a clinical case of treating the patient with injury of ulnar nerve using a “gold standard” of microsurgery, the autonerve grafting, and transplantation of autologous cells of stromal vascular fraction (SVF) of adipose tissue. Nerve defect in 5 cm was replaced by three autonerve grafts of the sural nerve, and then autologous cells of SVF in the fibrin glue were transplanted. 1 year after operation, the patient has resumed the function of bringing the fingers, there have not been any neurogenic strain, muscle atrophy of the hand, as well as restoration of sensitivity in the autonomous zone of innervation of the ulnar nerve has been observed.
Genes & Cells. 2014;9(3):307-311
pages 307-311 views

Description of methods for the detection of Helicobacter pylori in children with chronic gastroduodenal pathology

Faizullina R.A., Abdullina E.V., Akhmetov I.I.


Frequent detection of Helicobacter pylori infection among both adult and child population, as well as its relationship with the development of chronic gastroduodenal pathology field justified the need to search for sensitive and accurate diagnostic methods. This has stimulated the emergence of a variety of different methods for detecting Helicobacter pylori, which created for clinicians and specialists laboratory diagnosis difficulties in choosing the adequate and reliable ways to identify. A comparative analysis of methods for diagnosis of Helicobacter pylori infection in 151 children with chronic gastroduodenal pathology field, including 23 patients with duodenal ulcer, 97 patients with chronic gastroduodenitis and 31 conditionally healthy children of the same age. Breathing helic - test and polymerase chain reaction of gastric mucosa biopsies for the diagnosis of Helicobacter pylori infection have equally high sensitivity (94.8% and 93.6%) but low specificity (80.6% and 79.5%) and a high probability helicobakteriosis methods with negative results (19.4% and 18.4%).The analysis of the diagnostic value of the combination helic - test and polymerase chain reaction in the verification of Helicobacter pylori infection showed a higher sensitivity (98.3%) and specificity (93.5%) overall accuracy (96%) and the probability of a positive result helicobakteriosis (96.6%) and a low probability in a negative result (6.4%) methods. Therefore, for the primary diagnosis of Helicobacter pylori infection is better to use a combination of breathing helic -test and polymerase chain reaction biopsies gastric mucosa, reducing the proportion of false-positive and false-negative data, would increase the sensitivity and specificity of the methods.
Genes & Cells. 2014;9(3):312-315
pages 312-315 views

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