bluuebunny/crossref_metadata_2025 huggingface dataset:
Transplantation of hepatic stellate, genetically modified genes hgf and fgf4, stimulates differentiation of hepatocytes in rats after partial hepatectomy
Authors: Titova A. A , Sharipova E. I , Pevnev G. O , Mavlikeev M. O , Shafigullina A. K , Burganova G. R , Titova M. A , Rizvanov A. A , Gumerova A. A , Kiyasov A. P
Published: October 2024

Chronic liver diseases of different etiology are the problem of the modern medicine. Nowadays is held an active search for new methods of treatment for these diseases using stem cells and methods of gene-cell therapy. One of the cell types, thought to be progenitor cells of the liver, - hepatic stellate cells. Previously was shown the possibility of hepatic stellate cells to differentiate into the cells with hepatocytes phenotype after transplantation into the rats with partial hepatectomy. In this work we studied rat liver regeneration on the same model of liver damage after transplantation of the hepatic stellate cells, transduced with therapeutic genes of hepatocyte growth factor (HGF) and fibroblast growth factor 4 (FGF4) in combination with tracer gene of red fluorescent protein (RFP). The results of the survey demonstrated that transduction of the hepatic stellate cells with these genes facilitates engraftment of the transplanted cells and their further differentiation into hepatic direction, that was confirmed by a large number of hepatocytes, expressing tracer gene RFP, as well as a large number of α-fetoprotein positive hepatoblasts in compare to the animals, that had transplantation of cells, transduced only with tracer gene
Lymphocyte apoptosis in neonatal sepsis due to klebsiella
Authors: Khaertynov Kh. S , Boichuk S. V , Anokhin V. A , Ramazanov B. R , Rizvanov A. A , Khaiboullina S. F , Lubin S. A , Agapova I. V
Published: October 2024

The apoptosis of lymphocytes was examined in 10 infants with late neonatal sepsis induced by Klebsiella pneumoniae Most of the infants (70%) were preterm. Control group consisted of 7 healthy newborns. Apoptosis analysis was examined by counting the numbers of hypodiploid cells by using a propidium iodide (DNA staining procedure and flow cytometry analysis. We observed an elevated numbers of apoptotic cells in all cases of neonatal sepsis Median of lymphocyte apoptosis was 18 1% on day 5 of cell culture, whereas in control patients it was up to 7,8%. Absolute lymphopenia was observed in 30% of cases with neonatal sepsis. The activity of lymphocyte apoptosis was not associated with quantitative baseline of C-reactive protein (Spearman R = 0,48, p = 0,18). Acute phase of neonatal sepsis induced by Klebsiella pneumonia is associated with an increased apoptosis of peripheral blood lymphocytes
Parameters of single and summated contractions of skeletal muscles in vivo and in vitro
Authors: Eshpay R. A , Khairullin A. E , Karimova R. G , Nurieva L. R , Rizvanov A. A , Mukhamedyarov M. A , Ziganshin A. U , Grishin S. N
Published: October 2024

It is well known that modern non-invasive methods for registration of functional activity of skeletal muscles in vivo due to technical reasons reflect amplitude and temporal parameters of muscle contractions not better than in vitro approaches The aim of present study is to register and compare parameters of single and tetanic contractions of different types of skeletal muscles of warm-blooded animals in vitro and under minimally invasive surgery in vivo. It was found that single muscle contractions in vitro and in vivo indistinguishably close to each other, whereas frequency of tetanus fusion and waveform of tetanic contractions are different.
Application of CRISPR/Cas9 system for developing and studying cellular models of inherited disease
Authors: Valetdinova K. R
Published: October 2024

Currently, more than 4500 diseases classified as an inherited diseases. Most forms of human inherited diseases are caused by gene mutations. Given to the development of new methods genome editing, particularly with the use of CRISPR/Cas9 system, it is possible to artificially introduce precise mutations in the genome, as well as to correct gene mutations. It is thus possible to create a cellular model of human inherited diseases, as well as to carry out experiments for testing hypotheses and principles of gene and cell therapy. In this review, we consider studies that provide insight into the possibilities of CRISPR/Cas9 system for developing and studying cellular models of human inherited diseases.
Sepsis and apoptosis
Authors: Khaertynov Kh. S , Anokhin V. A , Boichuk S. V , Rizvanov A. A
Published: October 2024

The article provides an overview of the current knowledge about the immune response in sepsis. Sepsis occurs on a background of development as a systemic inflammatory response, and immunosuppression phenomena. Dysfunction of the immune system is one of the most important parts of the pathophysiology and compulsory process. One of the immunosuppression mechanisms in sepsis is lymphocyte apoptosis. The article describes the main activation pathway of this process including those in neonatal sepsis.
Methods of gene delivery and perspectives of their application in the gene therapy
Authors: Apartsin E. K , Knauer N. Yu
Published: October 2024

Gene therapy is believed to be among the most promising directions of the future medicine. Thus, the development of efficient and safe methods of the nucleic acid delivery to the target cells, tissues and organs ecomes of great current interest. This review summarizes recent data on the approaches for the gene delivery and discusses clinical aspects of the gene therapy.
Hepato-specific small-dispersioned matrix as the important component of implanted cell-engineering designs for an auxiliary liver
Authors: Onischenko N. A , Krasheninnikov M. E , Shagidulin M. Y , Bodrova M. M , Sevastjanov V. I , Gautier S. V
Published: October 2024

Biocompatible and biodegraded matrixes have already framed by the modern biomedical technologies however working out and application of tissue-specific matrixes remains an actual problem of the modern tissue engineering Aim of this work is to show that the technique, which is proposed by the authors for producing of small-dispersioned matrices of decellularization liver (DCL), is good for the making of cell-engineering designs of auxiliary liver The paper presents the technology of producing the small-dispersioned matrix of DCL and the results of using the light, phase- contrast and electron microscopy to characterize the biological properties of the made matrix The hepatospecific properties of the matrix have been studied by using a quantitative evaluation of the MTT-test results on the 5th day of separate cultivation on this matrix 4 types of cells (HepG2, renal epithelial cells, bone marrow MSCs and liver cells from allogeneic donors). On photographs of microscopic examination of the matrix particles of DCL it was seen their porous structure, on the surface of which the preserved conglomerates of native extracellular matrix molecules were presented At comparative study of adhesive properties of the Cytodex-3 matrix particles and the small-dispersioned matrix of DCL it was found out that both matrices had the ability to adhere different cell types, but the matrix of DCL had the ability to preserve of hepato-specific activity significantly expressed. Keeping of biocompatibility and hepato-specific properties by small-dipersioned matrices of DCL, produced on the proposed technology, allows to recommend them for the making of implantable cell-engineering designs of auxiliary liver.
Osteogenic potential of multipotent mesenchymal stromal cells from human exfoliated deciduous teeth before and after cryopreservation
Authors: Bukharova T. B , Leonov G. E , Galitsyna E. V , Vasilyev A. V , Vakhrushev I. V , Vikhrova E. B , Makhnach O. V , Goldstein D. V
Published: October 2024

The use of multipotent mesenchymal stromal cellsfrom human exfoliated deciduous teeth (SHED) to stimulate bone regeneration requires data on the influence of cryopreservation on the osteogenic differentiation capacity of these cells. SHED were subjected to cryopreservation. Before freezing and after thawing, cell cultures were exposed osteogenic differentiation with vitamin D3 or dexametasone and assessed for expression of the osteogenic markers osteocalcin, alkaline phosphatase, BMP-2 and RunX2 using real-time qPCR. Extracellular matrix (ECM) mineralization was evaluated by Alizarin red staining. Supplementation of osteogenic medium with vitamin D3 increased the expression of the osteogenic markers osteocalcin, alkaline phosphatase, BMP-2 and RunX2 as well as promoted an increase in the synthesis and mineralization of ECM in the cells both before and after cryopreservation. In the presence of vitamin D3 gene expression of alkaline phosphatase, BMP-2 and RunX2 after cryopreservation was higher than before freezing. Gene expression of osteocalcin, BMP2 RunX2 in osteogenic medium with vitamin D3was higher compared with dexamethasone for 14 days differentiation both before or after cryopreservation. The maintenance of SHED osteogenic differentiation potential after long-term cryopreservation provides a basis for banking of these cellsfor further auto- or allotransplantation.
Impact modified glial-derived neurotrophic factor (GDNF) for regeneration of epithelial and epithelial-stromal corneal defect in the experiment
Authors: Gavrilova N. A , Revischin A. V , Borzenok S. A , Komova O. J , Agammedov M. B , Tonaeva H. D , Ostrovsky D. S , Pavlova G. V
Published: October 2024

Objective is to study the effect of modified glial-derived neurotrophic factor (GDNF) on healing of epithelial and epithelial-stromal corneal lesions in mice C57BL / 6J. After corneal damage the instillations of supernatant conditioned by HEK293 cells expressed GDNF gene construction without pre- and pro- sequences were produced. For control, a medium conditioned by not transfected cells was used. We assessed the area of corneal epithelial defect and corneal erosion rate, developing after the epithelial defect. The immunohistochemical study using antibodies against cytokeratin 5/18, c-Met, collagen IV, phospho-ERK1/2, phospho-JNK1/2, Ki67, Bcl2, GAP43, TIMR-1, TGF-p, Bax, and MMP 9 was performed. The area of corneal epithelial defect in the eyes of experimental animals within one day after damage was smaller than in the control. Frequencies of corneal erosions formed in the eyes of experimental animals after damage was observed in 30-35% and 80-85% of cases in the experimental and control groups, respectively. Immunohistochemical studies using these antibodies showed that GDNF stimulated the proliferative activity of epithelial cells and keratinocytes, contributed to active migration and adhesion of epithelial cells, had anti-apoptotic and antifibrotic effects, took an active part in the formation of stromal nerve plexus. The results indicate the hopefulness of therapeutic application of the modified GDNF after corneal injury and the need for further research to develop and test methods for the therapeutic use of drugs on the basis of this neurotrophic factor.
Fascaplysinis a promising agent for the creation of new treatments' methods of glial brain tumors
Authors: Bryukhovetskiy I. S , Zhidkov M. E , Kudryavtsev I. V , Polevshikov A. V , Mishchenko P. V , Milkina E. V , Bryukhovetskiy A. S , Zaitsev S. V , Lyahova IA. A , Vikhareva V. V , Khotimchenko Yu. S
Published: October 2024

The effectiveness of glial brain tumors treatment remains low. Standard treatments do not provide a radical removal of tumor cells infiltrating the brain substance. One solution to this problem is to find new compounds with high anti-tumor activity and the methods' development of targeted delivery. Our attention was drawn to a group of compounds, which are based on pentacyclic system of pyrido[1,2-a: 3,4-b'] diindols. The most famous representative is fascaplysin. It is the first substance isolated from a marine sponge Fascaplysynopsis sp. The objective was to explore the characteristics and mechanisms of the cytotoxic and cytostatic effect of fascaplysin on C6 glioma cells in vitro, and to compare the effectiveness fascaplysin and temozolomide in vitro and in a targeted delivery to the tumor in vivo. We used techniques: mammalian cell cultures, high-performance robotic quantitative microscopy, confocal laser fluorescence microscopy, flow cytometry, modeling of glioblastoma in vivo, pharmacological testing, magnetic resonance imaging. It is shown that fascaplysin induces apoptosis in C6 glioma cells. At a concentration of 2 uMfascaplysin was more effective than the temozolomide. Decrease of concentration below 0.5 uM resulted to reduction of cytotoxic effects. The severity of cytostatic effect increased with increasing exposure time. Targeted delivery the fascaplysin to the tumor using transport capacity of stem cells increased the lifespan of rats and decrease in tumor size.
Features of interaction between hematopoietic stem and tumor cells of different lines in vitro
Authors: Milkina E. V , Mischenko P. V , Zaytsev S. V , Bryukhovetskiy I. S , Dyuyzen I. V , Bryukhovetskiy A. S , Khotimchenko Y. S
Published: October 2024

One of the promising directions of the brain tumors treatment connected using hematopoietic stem cells (HSC). The aim of this work: to study the features of the interaction between the human stem and tumor cells of different lines. The materials for the study were CD 34+-HSCs, U87 glioblastoma, lung cancer (A549), breast cancer (MCF-7). We used cell culture methods, the robotic monitoring of cellcell interactions, confocal laser microscopy, flow cytometry. It was shown that the mobility of HSCs to glioblastoma cells was higher than to cells of lung and breast cancer. The high mobility of HSCs combined with the ability to adhere to tumor cells and to share with them the contents of the cytoplasm. A direction of transport cytoplasm was depended from the tumor cell types and is a feature of the process. We believe that the described phenomenon can be one of the mechanisms of invasive growth and also indicates a high antitumor potential of HSCs.
Neural differentiation of patient-specific induced pluripotent stem cells from patients with a hereditary form of spinal muscular atrophy
Authors: Grigor'eva E. V , Valetdinova K. R , Ustyantseva E. I , Shevchenko A. I , Medvedev S. P , Mazurok N. A , Maretina M. A , Kuranova M. L , Kiselev A. V , Baranov V. S , Zakian S. M
Published: October 2024

Induced pluripotent stem cells (iPSCs) give the possibility for disease modeling, drug and toxicology screening and development of the new therapeutic approaches. Directed differentiation of iPSCs into specialized cell types represents a unique tool in order to study and model certain diseases, which affects specific type of cells, in vitro. One of the typical example of such diseases is spinal muscular atrophy, which is caused by mutations in the SMN1 gene (survival motor neuron 1 gene), leading to selective death of motor neurons. Patient-specific iPSCs were derived from the patient with a hereditary form of spinal muscular atrophy I type and expressed the markers of pluripotency (NANOG, TRA-1-60, SSEA4, OCT4, KLF4, MYC, REX1, and others). Spontaneous differentiation of the obtained cells resulted in the appearance of derivatives of the three germ layers: ecto-, meso- and endoderm. Neural differentiation showed the appearance of the early neural markers (PAX6, SOX2, NESTIN, TuJ1, PSA-NCAM), the late mature neural markers (MAP2, NF200, GFAP), and the mature motor neurons' markers (ISL1 and CHAT). Neurons derived from patient-specific iPSCs are perspective model for studying the features of the cells, which are altered in spinal muscular atrophy.
Pathogenetic parallels between native and bioprosthetic aortic valve calcification
Authors: Mukhamadiyarov R. A , Rutkovskaya N. V , Milto I. V , Vasyukov G. Yu , Barbarash L. S
Published: October 2024

There was a suggestion that pathological tissue mineralization is a universal, multifactorial, and cell-mediated process, regulated on genetic, biochemical and hormone levels. Structural deterioration of bioprosthetic heart valves is mainly caused by calcification of chemically modified xenogenic tissues. We performed this study with the aim to define parallels in degenerative strokes of native aortic valves and Ca-mediated primary tissue-insolvency of bioprosthetic aortic valves. We investigated calcified tissue samples of bioprosthetic aortic valves and calcified wings of aortic valves. (eight per each group). Functionally safe xenopericardiac mitral prosthetic heart valve, obtained at autopsy were included to investigate processes with prosthetic wings. Histological examination was carried out by hematoxylin and eosin and van Gieson staining on optical microscope AXIOImager A1. We observed mosaic structural alterations which characterized increasing of calcification process in all specimens. There were interactions between mineral deposits and valve tissues by direct contact with the cells and/or fibers of collagen and elastin. Cell composition in calcified native aortic valves was represented by fibroblasts, macrophages, multinucleated giant cells, lymphocytes, and neutrophils. Bioprosthetic heart valves also contained smooth muscle cells and capillaries. Results of our investigation represents that structures of calcified native and bioprosthetic heart valves are similar. This fact make us able to conclude that there are direct cells-mediated processes of biological material and xenogenic aortic valves degradation. Appearance of structurally safe cells in devitalized xenogenic prosthetic aortic valve caused by their functioning in recipient body. We suggest that the source of these cells can be the pluripotent progenitor cells, circulated in bloodstream and able to differentiate into cells of different phenotypes when they penetrated the collagen matrix of implanted prosthetic aortic valves. Based on the above analysis we can suggest that calcium degeneration of soft tissues is an universal pathologic process the basis of which have different mechanisms and the key one is cell-mediated mineralization.
Genome editing approach for generation of isogenic cell lines modelling Huntington's disease in vitro
Authors: Malakhova A. A , Sorokin M. A , Sorokina A. E , Malankhanova T. B , Mazurok N. A , Medvedev S. P , Zakian S. M
Published: October 2024

Huntington's disease is an autosomal dominant neurodegenerative disorder caused by trinucleotide repeats expansion mutation in the first exon of HTT gene. Neurodegenerative disorders are difficult to study due to limited material availability and late onset of the manifestation. In vitro cell models provide a good opportunity to study molecular mechanisms of the pathology. New genome editing approaches allow us to generate isogenic cell lines that can be useful for drug screening and studying mechanisms of molecular and cellular events triggered by certain mutation on an equal genetic background. The aim of our work was establishing a method to introduce expanded CAG repeat tract into HTT gene by homology directed repair. We used a genome editing tool based on CRISPR/Cas9 system to facilitate the process. A donor construct bearing 215 CAG trinucleotide repeats flanked with long homology arms was generated. The method was approved on the HEK293Phoenix cell line. Mutant clones were obtained. It was shown that mutant HTT protein is expressed in the HEK293 mutant clones and affect their viability and proliferation rate.
Genome editing on cell model of the genetic form of Parkinson's disease
Authors: Vetchinova A. S , Konovalova E. V , Volchkov P. Yu , Abramycheva N. Yu , Illarioshkin S. N
Published: October 2024

New technological approaches for modeling of neurodegenerative disorders play growing role in modern neurobiology. One of such technologies is targeted genome editing with the use of artificial nuclease systems (CRISPR/ Cas9, etc.), that allow to correct genetic defects on the cellular level. Especially promising is the use of genome editing technology on specialized neurons and induced pluripotent stem cells obtained by cell reprogramming from patients with hereditary forms of neurodegenerative diseases. Using CRISPR/Cas9 system we performed editing of genome of induced pluripotent stem cells obtained from fibroblasts of a patient with autosomal recessive form of Parkinson's disease carrying compound heterozygous mutations in the PARK2 gene. Genome editing allowed to restore normal nucleotide sequence in both alleles of the PARK2 gene, correcting exonic (del202-203AG) and intronic (IVS1+1G/A) mutations. Possibility of genome editing and further obtaining of normal dopaminergic neurons from induced pluripotent stem cells is important both for studies of molecular mechanisms of neurodegenerative disorders and for successful development of cell therapy methods for Parkinson's disease.
Optimization of teratoma formation assay
Authors: Kizilova E. A
Published: October 2024

Teratoma formation assay is necessary to estimate in vivo pluripotency of stem cells especially stem cell lines of human origin. Nevertheless convenient, valid and universal “standards” to analyze stem cell derived tumors have not been developed yet. New protocol for monitoring teratoma growth, morphological and histological analyzes of tumor samples is proposed in this paper. This protocol is oriented on review of tumors morphology and histology per se. The list-describer includes 17 obligate and 12 facultative diagnostic sell types and 7 diagnostic cell complexes. The protocol takes into account complicity and heterogeneity of teratoma structure and allows detect different morphological features of malignization process inside stem cell derived tumors in situ. The protocol was successfully applied for teratoma formation test which has been performed for 52 stem cell lines of different species origin (mouse, rat, аmerican mink and human). 326 stem cell derived tumors were completely described, reviewed and analyzed.
Current state of EDTA-associated pseudothrombocytopenia, importance in the pathogenesis of polymorphism in the gene of platelet receptor for fibrinogen
Authors: Polyakov A. S , Goncharova E. V , Bologov S. G , Zhogolev D. K , Bondarchuk S. V , Noskov Ya. A , Petrova O. R , Voronin S. V , Kolubaeva S. N
Published: October 2024

EDTA-associated pseudothrombocytopenia a laboratory phenomen associated with the widespread introduction into laboratory practice of hematology analyzers and standardized methods for blood samples preserving. High frequency pseudothrombocytopenia among healthy individuals (20%) and among patients with various pathological conditions (50%), and lack of control of the results, can cause a large amount of laboratory and diagnostic errors. Currently there is no reliable data on the clinical value pseudothrombocytopenia. Identify EDTA-pseudothrombocytopenia based on a comparison of the data automated hematological analysis with the results of microscopic examination platelets count or automated studies using alternative anticoagulants. Detection of somatic mutations in the platelet fibrinogen receptor gene (integrin gene polymorphism ITGB3-b: 1565 T>C) in a limited study - 63.2% (12 patients surveyed from 1 9) may serve as the beginning of a new direction in understanding the etiopathogenesis and clinical significance of pseudothrombocytopenia.
Cellular blood composition: hemogram healthy individuals in Saint Petersburg
Authors: Bondarchuk S. V , Tyrenko V. V , Mihaleva M. A , Yurkin A. K
Published: October 2024

Presents the results of a population-based study of the numerical values of the blood of the adult male population of St. Petersburg. Studied the frequency distribution of the major indicators of the investigated cohort hemogram for 7 years. Revealed a significant variability in cellular composition. Found that interval distribution of the number of leukocytes, hemoglobin, eosinophils etc falls outside of the bounds of reference standards indicators. The most frequently mentioned decrease in the number of platelets, red blood cells, decrease in hemoglobin. Less frequently mentioned decrease in the level of lymphocytes, platelets level increase above normal interval distribution. Systemic blood diseases have been detected in the study group. Deviations can be caused by physiological oscillations, plasticity of the adaptive capacity of the blood. Thus, the distribution of the population significantly healthy hemogram feature. Justified the extension of the reference indicators of cellular components of blood.
Comparison of cytogenetic damage in groups of patients with myeloproliferative neoplasms, depending on the presence or absence of V617F mutation in JAK2 gene
Authors: Kolubaeva S. N , Polyakov A. S , Kissel A. V , Myakoshina L. A , Viktorova N. A , Ushanov S. S , Titova A. A , Ivanov A. M , Noskov Ya. A , Bologov S. G , Voronin S. V
Published: October 2024

Myeloproliferative neoplasms are diseases characterized by clonal proliferation of myeloid bone marrow. At present, it describes a number of genetic markers myeloproliferative neoplasms, such as JAK2v617f. However the nature of violations in the progenitor cells is still not clear. In this study, a comparative analysis of cytogenetic damage in patients with myeloproliferative neoplasms with a mutation JAK2v617f and without this damage. The study showed that differences in the number and diversity of received clonal damage in both groups was not significant. Given the small number of patients with clonal cells, it can be assumed that cytogenetic analysis diagnostically significant.
Combined treatment of throphic ulcer of the heel using vacuum therapy with direct gene therapy: case report
Authors: Mullin R. I , Masgutov R. F , Salafutdinov I. I , Rizvanov A. A , Bogov A. A
Published: October 2024

In this article we describe a clinical case of successful treatment of neurotrophic ulcers of heel region using classical surgical techniques such as debridement and autodermoplastics combined with vacuum therapy and injection of double cassette expression plasmid encoding cDNAs of vascular endothelial growth factor and fibroblast growth factor 2. The outcome of therapy opens new horizons in the treatment of such diseases in short period of time.
Regenerative technologies in treatment of diabetic foot ulcers
Authors: Kalinin R. E , Suchkov I. A , Mzhavanadze N. D , Krylov A. A , Isaev A. A , Plaksa I. L , Deev R. V
Published: October 2024

Regenerative medicine is one of the new fast growing approaches to the treatment of a variety of diseases, including oncology and cardiovascular pathology, degenerative-dystrophic diseases of bones and joints, and other complex diseases including diabetes mellitus. Diabetes is regarded as one of the most dangerous non-infectious diseases in the world with a rising incidence that reaches pandemic proportions. Treatment of patients with diabetic foot ulcers including those with peripheral arterial disease is a challenging issue due to the increased risks for poor and lethal outcomes, which makes the social and economic burden of diabetes mellitus very high. The article presents state-of-the-art regenerative technologies in treatment of diabetic foot ulcers. Special attention is paid to gene and cell therapy.
Comparison of different methods of rat hepatic stellate cells isolation, labeling and transplantation
Authors: Shafigullina K. K , Trondin A. A , Burganova G. R , Titova A. A , Mavlikeev M. O , Sharipova E. I , Tabanakova A. V , Gazizov I. M , Kaligin M. S , Titova M. A , Rizvanov A. A , Gumerova A. A , Kiassov A. P
Published: October 2024

Nowadays more and more attention is turned to hepatic stellate cells (HSC) and their role in liver regeneration. Nonetheless there are several methodological questions, for example, methods of HSC isolation, their labeling and ways of transplantation. In this work we compared two different methods of HSC isolation: collagenase-pronase liver perfusion with further Histodenz density gradient centrifugation and method of Seglen for isolation of hepatocytes associated with HSC. We also analyzed diverse methods of cells labeling: membrane fluorescent labels PKH26 and with gene of green fluorescent protein (GFP), that could be get into the cells by electroporation, with chemicals like TurboFect or by adenovirus. Then cells were transplanted into rats in two ways: into lien and into system of portal vein. According to our results, we able to conclude that collagenase-pronase liver perfusion with further cells gradient centrifugation in Histodenz is better for HSC isolation than method of Seglen, the most optimal method for cells labeling is with adenovirus, expressing the GFP gene, for HSC transplantation - transplantation into system of portal vein.
Stem cell therapy of heart failure: clinical trials, problems and perspectives
Authors: Kochegura T N , Efimenko A Yu , Akopyan Zh A , Parfenova E E , Kochegura T N , Efimenko A Y , Akopyan Zh A , V EV M
Published: October 2024

Today cell therapy is considered as a new perspective approach in the treatment of different structural and functional changes in ischemic myocardium. In the present review we analyze last achievements in autologous stem and progenitor cells use for the regeneration of damaged myocardium at postinfarct heart failure, chronic ischemic cardiomyopathy and chronic heart failure.
Prospects of increasing the effectiveness of cardio-vascular gene and cell therapy: genetically modified cells
Authors: Shevchenko E K , Talitskiy A K , Parfenova E V , Shevchenko E K , Talitskiy A K , Parfyonova Y V
Published: October 2024

Many hopes for increasing the effectiveness of therapy for cardio-vascular diseases such as non optimal for cases of ischemic heart diseases, myocardial infarction, heart failure, critical limb ischemia are connected with progress in gene and cell therapy. Experimental studies strongly demonstrated effective angiogenesis and myocardial regeneration, blood flow restoration in ischemic limb by gene and cell therapy approaches. However the results of clinical studies of these methods are very modest. To overcome the insufficient effectiveness of these methods the genetic modification of stem and progenitor cells - an alliance of gene and cell therapy allowing to neutralize disadvantages and to enhance advantages of both methods - could be used. In this review we have provided an overview of pioneering research in the experimental application of genetically engineered cells in cardiovascular pathology. The possibility of using genetically modified progenitor cells of different origin for therapeutic angiogenesis, myocardial regeneration and creation of cardiac pacemakers is discussed.
The study of biocompatibility of heart value transplants devitalized by anticalcinosis treatment
Authors: Akatov B C , Muratov P M , Fadeeva I S , Sachkov A S , Britikov D V , Fesenko N I , Solov'ev V V , Chekanov A V , Akatov V S , Muratov R M , Fadeeva I S , Sachkov A S , Britikov D V , Fesenko N I , Soloviev V V , Chekanov A V
Published: October 2024

Earlier we have proposed method for anticalcinosis devitalization of heart valve transplants, which induced total cell death without elimination of cell debris from tissue matrix. It was shown in previous studies prevention of calcinosis in devitalized aorta transplants by the method of subcutaneous implantation in rats. In the present research we investigated biocompatibility of devitalized heart valve alio- and xenografts in the model of chronic implantation in descending aorta of dogs. It was revealed that viable and devitalized aorta grafts became acellular after 4-month implantation. Anticalcinosis divitalization of aorta allografts suppressed immune response, degeneration and calcinosis after their implantation in dogs. Xenografts after anticalcinosis treatment induced strong immune response resulted in fibrous reorganization of tissue matrix. According to the results obtained heart valve allografts devitalized by anticalcinosis treatment can be recommended for clinical trials.
The use of transplants containing multipotent mesenchymal stromal cells for the repair of the articular surface in experiment
Authors: Grigoryan A S , Deev R V , Kruglyakov P V , Bilibina A A , Sokolova I B , Pavlichenko N N , Polyntsev D G , Grigorian A S , Deev R V , Kruglyakov P V , Bilibina A A , Sokolova I B , Pavlichenko N N , Polyntsev D G
Published: October 2024

The effects of the allogeneous demineralized bone transplants, seeded with bone marrow-derived multipotent mesenchymal stromal cells CMSCsJ, on the healing of the damaged articular cartilage and the subchondral bone were studied. It was shown that the use of allogeneous as well as autologous MSCs on the demineralized bone transplants and also on the transplants combined with type I collagen gel promote the remodeling of the regenerating tissue and the recovery of the histotypic cartilaginous and osseous structures in comparison with the articular surface regeneration without any therapy or after the procedure of the mosaic chondroplasty. Nevertheless, the positive effects of cell therapy were not clinically prominent, which means most probably in that case that the generally accepted experimental model is not adequate in full measure, but the approach used is promising.
Effect of simultaneous expression of various usoforms of vascular endothelial growth factor VEGF and fibroblast growth factor FGF2 on proliferation of human umbilical cord blood cells HUVEC
Authors: Salafutdinov I I , Shafigullina A K , Yalvach M E , Kudryashova N V , Lagar'kova M A , Shutova M V , Kiselev S P , Masgutov R F , Zhdanov R I , Kiyasov A L , Islamov P P , Rizvanov A A , Salafutdinov I I , Shafigullina A K , Yalvac M E , Kudryashova N V , Lagarkova M A , Shutova M V , Kiselev S L , Masgutov R E , Zhdanov R I , Kiyasov А Р , Islamov R R , Rizvanov A A
Published: October 2024

Transplantation of stem cells, progenitor and differentiated cells is currently actively investigated as an approach for correcting degenerative diseases. However the mechanisms of therapeutic effect of cell therapy remains poorly understood. Nowadays a theory of paracrine stimulation of regeneration processes by transplanted cells through secretion of trophic and growth factors gains a particular popularity. Genetic modification of cells prior to transplantation allows efficient expression and targeted delivery of various therapeutic factors. The use of plasmid expression vectors are considered to be one of the safest and promising approaches for genetic modification of cells. We report generation of genetic constructs based on expression plasmid pBudCE4.1 [containing two independent expression cassettes] encoding different isoforms of human vascular endothelial growth factor (VEGFJ and basic fibroblast growth factor CFGF2). Expression of recombinant proteins was confirmed by immunoblotting and immunochistochemistry. We demonstrated that genetic modification of model cell line HEK293 by our expression plasmids resulted in paracrine stimulation of human umbilical vein endothelial cells CHUVECJ proliferation in vitro.
Can Perisinusoids! Cells Be Regional Stem (Progenitor) Cells of the Liver?
Authors: Gumerova A A , Kiyasov A P
Published: October 2024

Regenerative medicine is one of quickly developing and promising areas of medicine in which there is essentially new approach to restoration of damaged organs by stimulation and Cor) use of stem (progenitor) cells for acceleration of regeneration. To realize this approach, it is necessary to know: what are stem cells? What are regional stem cells? What are their phenotype and potencies? Stem cells are already identified for a number of organs and tissues (epidermis, skeletal muscle) and their niche is determined. However liver, the organ whose regenerative abilities are known since antique times, has not opened its main secret yet - the secret of a regional stem cell. In this review on the basis of our own and literature data we discuss our hypothesis that perisinusoidal stellate liver cells can be liver stem cells.
Suppression of calcification of heart valve transplants by their devitalization
Authors: Akatov V S , Fesenko N I , Soloviev V V , Fadeeva I S , Chekanov A V , Muratov R M , Britikov D V , Sachkov A S
Published: October 2024

Suppression of calcinosis and degeneration of heart valve transplants is important aim for improving their durability. It was shown previously that well-known methods of decellularisation including treatments with enzymes or some detergents or with hypotonic solution didn't suppress calcification of implanted aorta. In the present study we showed potential in prevention of calcinosis by a treatment, which induced death of donor cells without their total destruction before implantation. This method of anticalcinosis devitalisation is based on a hypothesis of mechanism of calcinosis initiation in heart valve transplants, which was earllier proposed in our group.
Novel plasmid constructs with angiogenic growth factors genes - human VEGF, HGF and angiopoietin-1 for therapeutic angiogenesis
Authors: Makarevich P I , Shevelev3 A Ya , Rybalkin I N , Kashirina N M , Lipatova L N , Tsokolaeva Z I , Shevchenko E K , Beloglazova I B , Boldyreva M A , Rubina К A , Vlasik T N , Parfyonova Ye V
Published: October 2024

Efficacy of critical limb ischemia gene therapy can be improved by application of novel plasmid vectors with higher transgene expression. The goal of this study is to evaluate in vitro and in vivo expression of angiogenic growth factors after gene transfer using a novel plasmid vector PC4W. Plasmid constructs with genes of human VEGF185 CpC4W-hHGFopt), HGF CpC4W-hHGFopt) and angiopoietin-1 (pC4W-hAng-1optJ were tested in vitro in a HEK293T cell culture. Cells were subjected to calcium-phosphate transfection and conditioning medium samples were assayed for transgene levels using Western blot and ELISA. Results were compared with commercially available pcDNA3 based vectors encoding the same growth factors. Reverse transcription PCR was used to assay transgene expression in BALB-c mice ischemic muscle. ELISA and Western blotting data suggest that PC4W based constructs give a higher protein output of about 2-2,5 fold compared with pcDNA3 based plasmids. Optimization of nucleotide sequence in growth factors cDNA results in additional increase in transgene expression. RT-PCR data shows that expression of human HGF persists in murine ischemic skeletal muscle up to 14 days after gene transfer. Our results indicate that novel plasmid constructs for angiogenic growth factors expression have a good efficacy in vitro and in vivo and can be used for VEGF1B5, HGF and angiopoietin-1 expression in human cell culture and in experimental animals' tissue. At the moment all developed constructs pass through a series of experiments in animal ischemia models and will be used for combined gene therapy development.
Effective Transduction of Human Adipose Stromal Cells by a Recombinant Adeno-Associated Virus
Authors: Shevchenko E K , Makarevich P I , Tsokolaeva Z I , Ratner E I , Parfionova E V
Published: October 2024

Despite the initial indications of positive therapeutic effects in cell therapy there are still limitations in numbers of autologous cell populations available without significant ex vivo expansion. Autologous adipose stromal cells CASCJ transplantation due to sufficient cell numbers, their multipotency and the ability to secret angiogenic growth factors may become an alternative tool to treat cardiovascular diseases. In this study we Investigated the ability to efficiently transfer gene into such cells using plasmid and recombinant adeno-associated virus (rAAVJ. Human ASC were isolated from adipose tissue obtained from different donors during surgical operations. Low passaged cells were transduced using gene delivery system CStratagene] based on recombinant adeno-associated virus (rAAVJ serotype 2 encoding human vascular endothelial growth factor (VEGF) or green fluorescent protein (GFP). Transduction efficiencies and transgene expression level in ASCs were analyzed by quantitative flow cytometry and ELISA. ASC population was analysed for heparan sulfate proteoglycan expression, the main cellular AAV binding receptor. It was found that 55-65% of human ASC population express this receptor. The efficiency of ASC transduction using AAV delivery system was found to be 60+7%. GFP expression was visible during a month. Relative to control, cells transduced by VEGF rAAV vector increased VEGF secretion level by at least 20-30 fold as compared to unmanipulated ASC. Recombinant adeno-associated virus provides efficient tools for ex vivo modification of human ASCs.
An Experience in Comprehensive Treatment of Hepatic Cirrhosis with Fetal Tissues
Authors: Pischkin S A , Pirogova I Yu
Published: October 2024

Treatment of complications of portal hypertention at liver cirrhosis also includes as preventive maintenance of varieces bleedings of a gullet various ways, and as treatment of hepatocellular insufficiency. The result of complex treatment with application of surgical methods and to stimulation of liver regeneration transplantation fetal tissue for correction of hepatocellular insufficiency are submitted.
Dinâmicas territoriais e sustentabilidade
Authors: Oliveira Thais , Klauck Samuel
Published: October 2024

O artigo "Dinâmicas territoriais e sustentabilidade: o papel do patrimônio cultural no Parque Nacional do Iguaçu" tem como objetivo explorar as interações entre patrimônio cultural, ambiente natural e sociedade dentro do contexto das áreas protegidas. A pesquisa foca no Parque Nacional do Iguaçu, analisando como a percepção e o reconhecimento dos valores culturais contribuem para a conservação ambiental. A metodologia subsidia-se em ensaio teórico, usando revisão de literatura com abordagem interdisciplinar, abrangendo áreas como geografia, cultura, antropologia e urbanismo. Os resultados destacam a importância dos valores culturais na eficácia das estratégias de conservação e no bem-estar das comunidades envolvidas, revelando que a gestão sustentável do parque pode beneficiar-se da integração de narrativas culturais. A pesquisa promove discussões sobre a coexistência sustentável e a conscientização pública, mostrando que a valorização do patrimônio cultural pode fortalecer os laços sociais e enriquecer a experiência dos visitantes, além de proporcionar uma compreensão mais rica e conectada do patrimônio natural e cultural global.
Effective governance and the doctrine of “total defence” as factors of state stability in wartime
Authors: Cherlenіak Ivan , Tokar Marian
Published: October 2024

In wartime, effective governance and the doctrine of “total defence” are critical factors that ensure the stability, security, and sovereignty of the state. The purpose of this study was to investigate the aspects of the effectiveness of management of the national security and defence system of the state. Analytical modelling of the systemic components of Ukraine’s resilience in wartime through the lens of the total defence doctrine and the concept of social self-organisation was used. The study investigated the barriers and difficulties in preparing the public authorities of Ukraine for effective management of resistance to the threats of hybrid warfare and threats of a full-scale high-intensity war along the entire line of the state border of Ukraine with the Russian Federation. The study analysed the specific features of the lowintensity hybrid war of 2014-2021 and the specific features of the initial stage of the high-intensity war of 2022. The study considered the achievements, omissions, shortcomings, and mistakes of public administration bodies and official systems in the first stages of the full-scale invasion of the Russian occupation forces into the cities and villages of Ukraine. It was shown that as a result, the Ukrainian defence forces were unable to organise an effective response to the largescale invasion of Russian troops in a prompt and complete manner. The study analysed the principles of organisation of the total defence system by public administration bodies of the Scandinavian and Baltic States. It was proved that at the present stage of the country’s defence, the concept of strengthening the resilience of the country’s social, economic, and military potential should be the most expedient. It was substantiated that in a high-intensity positional war, effective public administration should be based on the concept of resilience, which is systematically linked to the doctrine of total defence of Ukraine. The findings of this study can be used by governments and civil society institutions for management and coordination strategies for agencies in times of war
Challenges of balanced development of regions in the context of centralisation, the case of Ireland
Authors: Chulaievska Maryna
Published: October 2024

The relevance of this article topic lies in assessing the impact of centralization of governance powers on ensuring balanced regional development, which is crucial for shaping effective regional policy and reducing socioeconomic disparities. The aim of this research was to analyze the challenges of achieving balanced regional development in the context of centralization of authority, using Ireland as a case study. During the study, regulatory acts, official materials of government bodies, results of sociological research, and scientific publications were analyzed using historical, comparative, and predictive methods of inquiry with an interdisciplinary approach. Analysis of sources from different historical periods allowed elucidating the evolution of phenomena and concepts under investigation. The concepts of balanced development of regions, development of metropolises, and sectoral development of the economy are contrasted in the article. The negative effects of strong centralisation, as well as its advantages for the development of the country are unpacked. The impact of the European Union's policy on the socio-economic growth of Ireland, aimed at reducing the level of centralization of decision-making, building local self-government, and reducing regional disparities, is highlighted. Based on these researches the possible impact of centralisation in Ukraine on post-war recovery and development was analysed and conclusions about the advantages, disadvantages and possibilities of applying the Irish experience in Ukraine in the context of conflict and post-war reconstruction is made. The possible development of public administration and local self-government in Ukraine is projected, taking into account the experience of Ireland. The approaches to regional development, which assume that the country's socio-economic development can be better ensured by reducing regional disparities, and approaches to the development of agglomerations and metropolises as a basis for the prosperity of the national economy is analysed. The findings of the study can be valuable for substantiating further development of decentralisation and reducing the level of forced centralisation that has emerged due to martial law, as well as for defining a conceptual vision for the reconstruction of the territory and regions of Ukraine