Increasingβ-hexosaminidase A activity using genetically modified mesenchymal stem cells

GM2 gangliosidoses are a group of autosomal-recessive lysosomal storage disorde rs.These diseases result from a deficiency of lysosomal enzymeβ-hexosaminidase A(HexA),which is responsible for GM2 ganglioside degradation.HexA deficiency causes the accumulation of GM2-gangliosides mainly in the nervous system cells,leading to severe progressive neurodegeneration and neuroinflammation.To date,there is no treatment for these diseases.Cell-mediated gene therapy is considered a promising treatment for GM2 gangliosidoses.This study aimed to evaluate the ability of genetically modified mesenchymal stem cells(MSCs-HEXA-HEXB)to restore HexA deficiency in Tay-Sachs disease patient cells,as well as to analyze the functionality and biodistribution of MSCs in vivo.The effectiveness of HexA deficiency cross-correction was shown in mutant MSCs upon intera ction with MSCs-HEXA-HEXB.The results also showed that the MSCs-HEXA-HEXB express the functionally active HexA enzyme,detectable in vivo,and intravenous injection of the cells does not cause an immune response in animals.These data suggest that genetically modified mesenchymal stem cells have the potentials to treat GM2 gangliosidoses.

Molecular and cellular changes in the post-traumatic spinal cord remodeling after autoinfusion of a genetically-enriched leucoconcentrate in a mini-pig model

Post-traumatic spinal cord remodeling includes both degenerating and regenerating processes,which affect the potency of the functional recovery after spinal cord injury(SCI).Gene therapy for spinal cord injury is proposed as a promising therapeutic strategy to induce positive changes in remodeling of the affected neural tissue.In our previous studies for delivering the therapeutic genes at the site of spinal cord injury,we developed a new approach using an autologous leucoconcentrate transduced ex vivo with chimeric adenoviruses(Ad5/35)carrying recombinant cDNA.In the present study,the efficacy of the intravenous infusion of an autologous genetically-enriched leucoconcentrate simultaneously producing recombinant vascular endothelial growth factor(VEGF),glial cell line-derived neurotrophic factor(GDNF),and neural cell adhesion molecule(NCAM)was evaluated with regard to the molecular and cellular changes in remodeling of the spinal cord tissue at the site of damage in a model of mini-pigs with moderate spinal cord injury.Experimental animals were randomly divided into two groups of 4 pigs each:the therapeutic(infused with the leucoconcentrate simultaneously transduced with a combination of the three chimeric adenoviral vectors Ad5/35‐VEGF165,Ad5/35‐GDNF,and Ad5/35‐NCAM1)and control groups(infused with intact leucoconcentrate).The morphometric and immunofluorescence analysis of the spinal cord regeneration in the rostral and caudal segments according to the epicenter of the injury in the treated animals compared to the control mini-pigs showed:(1)higher sparing of the grey matter and increased survivability of the spinal cord cells(lower number of Caspase-3-positive cells and decreased expression of Hsp27);(2)recovery of synaptophysin expression;(3)prevention of astrogliosis(lower area of glial fibrillary acidic protein-positive astrocytes and ionized calcium binding adaptor molecule 1-positive microglial cells);(4)higher growth rates of regeneratingβIII-tubulin-positive axons accompanied by a higher number of oligodendrocyte transcription factor 2-positive oligodendroglial cells in the lateral corticospinal tract region.These results revealed the efficacy of intravenous infusion of the autologous genetically-enriched leucoconcentrate producing recombinant VEGF,GDNF,and NCAM in the acute phase of spinal cord injury on the positive changes in the post-traumatic remodeling nervous tissue at the site of direct injury.Our data provide a solid platform for a new ex vivo gene therapy for spinal cord injury and will facilitate further translation of regenerative therapies in clinical neurology.

Mesenchymal stem cells and the neuronal microenvironment in the area of spinal cord injury

Cell-based technologies are used as a therapeutic strategy in spinal cord injury(SCI). Mesenchymal stem cells(MSCs), which secrete various neurotrophic factors and cytokines, have immunomodulatory, anti-apoptotic and anti-inflammatory effects, modulate reactivity/phenotype of astrocytes and the microglia, thereby promoting neuroregeneration seem to be the most promising. The therapeutic effect of MSCs is due to a paracrine mechanism of their action, therefore the survival of MSCs and their secretory phenotype is of particular importance. Nevertheless, these data are not always reported in efficacy studies of MSC therapy in SCI. Here, we provide a review with summaries of preclinical trials data evaluating the efficacy of MSCs in animal models of SCI. Based on the data collected, we have tried(1) to establish the behavior of MSCs after transplantation in SCI with an evaluation of cell survival, migration potential, distribution in the area of injured and intact tissue and possible differentiation;(2) to determine the effects MSCs on neuronal microenvironment and correlate them with the efficacy of functional recovery in SCI;(3) to ascertain the conditions under which MSCs demonstrate their best survival and greatest efficacy.

Combination of epidural electrical stimulation with ex vivo triple gene therapy for spinal cord injury:a proof of principle study

Despite emerging contemporary biotechnological methods such as gene-and stem cell-based therapy,there are no clinically established therapeutic strategies for neural regeneration after spinal cord injury.Our previous studies have demonstrated that transplantation of genetically engineered human umbilical cord blood mononuclear cells producing three recombinant therapeutic molecules,including vascular endothelial growth factor(VEGF),glial cell-line derived neurotrophic factor(GDNF),and neural cell adhesion molecule(NCAM)can improve morpho-functional recovery of injured spinal cord in rats and mini-pigs.To investigate the efficacy of human umbilical cord blood mononuclear cells-mediated triple-gene therapy combined with epidural electrical stimulation in the treatment of spinal cord injury,in this study,rats with moderate spinal cord contusion injury were intrathecally infused with human umbilical cord blood mononuclear cells expressing recombinant genes VEGF165,GDNF,NCAM1 at 4 hours after spinal cord injury.Three days after injury,epidural stimulations were given simultaneously above the lesion site at C5(to stimulate the cervical network related to forelimb functions)and below the lesion site at L2(to activate the central pattern generators)every other day for 4 weeks.Rats subjected to the combined treatment showed a limited functional improvement of the knee joint,high preservation of muscle fiber area in tibialis anterior muscle and increased H/M ratio in gastrocnemius muscle 30 days after spinal cord injury.However,beneficial cellular outcomes such as reduced apoptosis and increased sparing of the gray and white matters,and enhanced expression of heat shock and synaptic proteins were found in rats with spinal cord injury subjected to the combined epidural electrical stimulation with gene therapy.This study presents the first proof of principle study of combination of the multisite epidural electrical stimulation with ex vivo triple gene therapy(VEGF,GDNF and NCAM)for treatment of spinal cord injury in rat models.The animal protocols were approved by the Kazan State Medical University Animal Care and Use Committee(approval No.2.20.02.18)on February 20,2018.

Gene-modified leucoconcentrate for personalized ex vivo gene therapy in a mini pig model of moderate spinal cord injury

We previously demonstrated that gene-modified umbilical cord blood mononuclear cells overexpressing a combination of recombinant neurotrophic factors are a promising therapeutic approach for cell-mediated gene therapy for neurodegenerative diseases,neurotrauma,and stroke.In this study,using a mini pig model of spinal cord injury,we proposed for the first time the use of gene-modified leucoconcentrate prepared from peripheral blood in the plastic blood bag for personalized ex vivo gene therapy.Leucoconcentrate obtained from mini pig peripheral blood was transduced with a chimeric adenoviral vector(Ad5/35 F)that carried an enhanced green fluorescent protein(EGFP)reporter gene in the plastic blood bag.The day after blood donation,the mini pigs were subjected to moderate SCI and four hours post-surgery they were intravenously autoinfused with gene-modified leucoconcentrate.A week after gene-modified leucoconcentrate therapy,fluorescent microscopy revealed EGFP-expressing leucocytes in spinal cord at the site of contusion injury.In the spleen the groups of EGFP-positive cells located in the lymphoid follicles were observed.In vitro flow cytometry and fluorescent microscopy studies of the gene-modified leucoconcentrate samples also confirmed the production of EGFP by leucocytes.Thus,the efficacy of leucocytes transduction in the plastic blood bag and their migratory potential suggest their use for temporary production of recombinant biologically active molecules to correct certain pathological conditions.This paper presents a proof-of-concept of simple,safe and effective approach for personalized ex vivo gene therapy based on gene-modified leucoconcentrate autoinfusion.The animal protocols were approved by the Kazan State Medical University Animal Care and Use Committee(approval No.5)on May 27,2014.

Mechanisms of change contraction of function of the muscles <i>in vitro</i>at allergic

In this work, mechanisms of influence of protein sensibility of an organism on contractile function of the isolated skeletal muscles of the mouse—“fast”—musculus extensor digitorum longus, “mixed”—musculus diaphragma and “slow”— musculus soleus are investigated. It is shown that at a protein sensitization all “fast”, “mixed” and “slow” skeletal muscles change the contractile properties. The vector of these changes for muscles with a various phenotypes carries opposite character. Force of the reduction caused carbacholine at a “slow” and “mixed” muscles increase, at “fast”—decreases. A vector of change of force of reduction on carbacholine at protein sensitization at these skeletal muscles correlates with changes of non-quantum secretion acetylcholine in a zone of a trailer plate. Opposite changes of functional properties of “fast” and “mixed” muscles and “slow” muscles of a shin of the mouse at protein sensitization are caused by dynamics cholinoceptive processes of excitation of membrane muscular fibers. It comes out with the assumption, that change of the contraction functions of skeletal muscles at protein sensitization is caused by changes of cholinoceptive processes of excitation of a membrane of muscular fibers, and other changes in system of electro-mechanical interface.

Filtering Two-Phase Medium in Pipes and Channels on the Entrance Part of Flow

Processes of filtering two-phase media in filtration devices play an important role in various industries. Significant role in the process of filtering is the initial section of flow, which defines the basic parameters： the profile and value of the velocity, pressure gradients, concentration and dispersion of sediment particles, etc. The problem is solved by the method of surfaces of equal cost, the results enabled to establish the influence of the input section on the filtering process.

5'-三磷酸腺苷对前列腺素F_(2α)子宫效应的增效作用

OBJECTIVE: To investigate the interaction of exogenous adenosine 5′ - triphosphate (ATP), a P2 receptor agonist, with prostaglandin F2α .(PGF2α ) on pregnant women in labor as well as on isolated human pregnant uterus preparations. METHODS: For an in vitro study, myometrial samples were obtained from 27 women undergoing elective cesarean delivery at term. Concentration- response relationships for ATP (10- 8 - 3 × 10- 4 mol/L), PGF2α .(10- 9 - 10- 5 mol/L), and their combination were obtained by using routine pharmacological organ bath technique. An in vivo study was performed with 34 pregnant women with dysfunctional abnormalities of the active stage of labor who were randomly allocated into 2 study groups. The women in the control group (18 patients) received intravenous prostaglandin F2α at an initial rate of 7.5 μ g/min, whereas the women in the ATP group (16 patients) received prostaglandin F2α concomitantly with ATP (0.45 nmol/min, intravenously). RESULTS: Adenosine 5- triphosphate at concentrations of 10- 6- 3 × 10- 4 mol/L and PGF2α at concentrations of 10- 8 - 10- 5 mol/L caused concentration- dependent contractions of isolated smooth muscle preparations of the human pregnant uterus. At concentrations of 10- 6 mol/L and below, ATP had no effects on mechanical activity of the isolated uterus, but at concentrations of 10- 7 mol/L and 10- 6 mol/L, it significantly potentiated the contractile responses of the uterus induced by PGF2α .(P < .05, 2- way analysis of variance). Patients receiving intravenous infusion of ATP as a supplement to PGF2α . treatment, compared with those without ATP, had a significantly shorter interval from the start of the treatment to full cervical dilatation (3.31 ± 1.49 hours and 4.67 ± 1.11 hours in ATP and control groups, respectively; P = .014, Wilcoxon Mann- Whitney test). The total dose of prostaglandin received was significantly lower in the ATP group than that of controls (1,489.8 ± 699.9 μ g and 3 394.2± 1,951.9 μ g, respectively; P = .003, Wilcoxon Mann- Whitney test). No side effects of ATP treatment were observed during or after infusion. CONCLUSION: Adenosine 5′ - triphosphate potentiates effects of PGF2α .on pregnant human uterus in vitro and in vivo and thus could be a useful supplemental drug to increase uterine contractility at labor.

Highly effective sorbents obtained by treating agrowaste products in cold plasma

Sorbents are widely used in purification of various liquids and gases, offering a universal means for wastewater and air cleaning. The most promising sorbents are those obtained from agrowaste products such as rice or buckwheat husk. Processing of husk in cold plasma modifies the composition, structure and surface properties of the raw material and raises its porosity, thus substantially improving the sorption properties of the final product.Husk as a raw material for producing sorbents has the following advantageous properties: (1) appropriate chemical composition; (2) low cost; (3) high dispersity, due to which there is no need in any special treatment of the material prior to its exposing to plasma; (4) scaly structure and developed porous surface ensuring a high surface-to-volume ratio. The best properties are displayed by the sorbents obtained in cold plasma under reduced pressures of 53.2 Pa. The raw material traverses the region occupied by the plasma and, as it does so, it gets heated up to a temperature of 250 - 350 ℃. The whole process involves two stages: combustion of the raw material and modification of its properties under the action of the plasma. The combustion proceeds due to the oxygen contained in the starting material. During the combustion, the hydrogen contained in the starting material and some part of the carbon also burn out. The resultant scaly sorbent is accumulated in a cooler. The scales are black; they range in sizes from 1mm to 5 mm. The sorbents obtained are remarkable for their useful properties and outperform most of the traditional sorbents used in modern industry. The starting materials are inexpensive, and their resources are almost unlimited. The sorbents have rather a low production cost (1.8-2.5 $/kg). The sorbents can be used for cleaning hydrosphere from water pollutants on a large scale. The degree of cleaning water surface from oil products with sorbents was a subject of investigation. The highest degree of purification the sewages up to 96.6% was observed in those cases where sorbents obtained from rice and buckwheat husk were used. Another advantageous feature of the sorbents stems from the practical good that can be benefited from their utilization. The sorbents saturated with pollutants can be used as fuel or in producing asphalt concrete. The sorbents may prove useful in pharmaceutical and food industries. Most advantageous conditions for producing such sorbents are available in rice-growing countries such as China, India, Japan and Korea.

Application of cold plasma technology in fiber-reinforced composite materials

A study is presented concerning a cold plasma technique for improving the bondability of high-strength high-modulus multi-filament polyethylene fibers to polymer matrices and the fibers impregnation with the objective to fabricate composite materials (CMs). Strong bonding between the matrixes and reinforcing fibers during the production of composites appears in the case if interaction is chemical. The value of the activation energy of the chemical interaction for very high performance polyethylene fiber was estimated. It was 1.14 eV. This allows using the cold plasma technique for producing CMs. In order to understand the effect of cold plasma treatment treated and untreated fibers were used to fabricate CMs. The strong bond between the matrix and plasma-activated fibers affects both the properties and failure mode of composite. The properties and failure modes were compared to those of CMs reinforced with untreated fibers. After plasma treatment the properties of CMs are increased. CMs are broken as a unit whole under tension. The ideas of the activating the fibers by cold plasma treatment above the activation energy of the chemical interaction may be extended over other types of the fibers and matrices to produce new types of fiber-reinforced composite materials with high physicomechanical indices.

Team sport, power, and combat athletes are at high genetic risk for coronavirus disease-2019 severity

Dear editor,As government restrictions put in place to slow the acceleration of the coronavirus disease-2019(COVID-19)pandemic start to ease,many people,including elite athletes,will begin to return back to their normal daily activities.Although the majority of risk factors for severe COVID-19-hypertension,respiratory system disease.

Development of a digital zenith telescope for advanced astrometry

Like other optical astrometric techniques,the Photographic Zenith Tube(PZT) has played a key role in the past observations of the Earth rotation,and it also has a potential to be applied to several other observations by taking advantage of automatic observations with self compensation of tilt of the tube.We here propose In-situ Lunar Orientation Measurement(ILOM) to study lunar rotational dynamics by direct observations of the lunar rotation from the lunar surface by using a small telescope like PZT with an accuracy of 1 milli-seconds of arc(1 mas) in the post-SELENE mission.Our second application is to obtain local gravity field on the Earth by combining deflection of the vertical measured by PZT and the position measured by Global Positioning System(GPS) or Global Navigation Satellite System(GNSS).The accuracy required for this purpose is not as strict as ILOM.We have already developed a Bread Board Model(BBM) of the telescope for ILOM and made some experiments in order to know the performance of the driving mechanism under a similar condition to the lunar environment showing high vacuum,large temperature change and dusty condition.We have also shown that it is possible to correct the effects of uniform temperature change upon the optical system by using a simple model with an accuracy of better than 1 mas.This model has the potential to attain the accuracy of 1 mas,based on the results of the experiments and the simulations.

Lunar project ILOM:application of the analytical theory of Lunar physical libration for the simulation of star observations

This study briefly describes the targets and problems of the future Japanese project In situ Lunar Orientation Measurement(ILOM),which is planned for the year 2017.One of the important parts of the project is to place a small optical telescope on the Lunar surface with the purpose to detect the Lunar physical libration with unprecedented accuracy 0.001 arcsec.At the present stage of research the computer simulation of future observations is going on,aiming to determine the moments of transition of a star through the first meridian and the polar distance of the star.Rotation of the Moon is being calculated under the analytical theory developed in the frame of a theme of the grant.A list of stars brighter than 12 m,whose coordinates are close to the Lunar precession pole motion,was constructed on the basis of several star catalogues.On average,for each moment of observation in the field of view of the telescope(1°) there are approximately 20-25 stars.Analyses of simulated stellar tracks observable from the Lunar surface(in a polar zone) reveal the significant difference from daily parallels of stars in comparison with the Earth.During one Lunar "day" equal 237 terrestrial days,a star moves on a spiral.However,depending on a longitude of a star,these spirals can be untwisted or twisted.In the latter case a star can describe a loop in the sky of the Moon during the period of supervision.Such an unusual astrometric phenomenon combined with the slow rotation of the Moon is compared with the Earth and the fast precession motion of the Lunar pole(in comparison with the precession motion of a terrestrial pole).

Structure of low-head two-phase flow in flat nozzlesduring outflow of the liquid into rarefied medium

Outcomes of experimental researches of the low-pressure adiabatic flow of the boiling liquid through two-dimensional Laval nozzles in a vacuum atmosphere were adduced.Requirements of critical conditions of flow were determined.Structural forms of a stream were investigated and their connection with crisis of flow was shown.It was established periodic non-stationary macrostructures of a stream which was stipulated by the rotational gear of origin of a vapor phase.