EFFECT OF DNA CONSTRUCTIONS ELECTROPORATION ON DENDRITIC CELLS

Today transfection of mammalian cell with DNA or RNA construction is the only method for delivering programmed information into the cell nucleus. Electroporation is most commonly used method of transfection in experiments with dendritic cell. The aim of electroporation is to permeabilize the membrane by passing electric impulse through the cell. Due to the increase permeability of the membrane chance DNA or RNA construction getting inside into the cell is increased, wherein survival of the cells is decreased.In the study male mice C57Bl/6 line 2-4 months old were used. From femur bones was isolated 20 × 106 bone marrow cells, which were cultured in 20 mL of complete RPMI-1640 for 7 days. To generate dendritic cells from BM cells, 100 ng/mL of rmFlt3-L was added to culture media at day 0. After 7 days of cultivation, the cell cultures were electroporated with control noncoding plasmids p5 (EP P5) or pmaxCCR9 encoding mouse chemokine receptor CCR9 (EP CCR9). The controls were cell cultures electroporated without any plasmids (mock EP) and cell cultures without electroporation (none EP). 5 × 105 cells were electroporated and resting for 10 minutes. After 10 minutes, cells were harvested and seeded into 24-well plates in 1 mL of culture medium and conditioning medium (1:1). Then, 50 ng/mL of Flt3-L was added to each well. The next day, transfected cells were collected and used for flow cytometry, qRT-PCR analysis.It was found that after electroporation in the groups mock EP, EP P5, EP CCR9 relative amount of live CD11c+ dendritic cells was significantly less than in the non EP group. Moreover, in the EP P5 and EP CCR9 groups the frequency of live CD11c+ dendritic cells was significantly less than in the mock EP group. Expression of the CD86 marker in the EP P5 and EP CCR9 groups was significantly higher than in the non EP and mock EP groups. Expression of the I-AB(MHCII) in the EP CCR9 group on cDC2s was significantly higher than in the non EP group. On plasmacytoid DCs (pDCs) and conventional type 2 DCs (cDC2s) in the EP CCR9 group expression of CCR9 was significantly higher than in the non EP group. Therefore, in this study, we demonstrated the effectiveness of electroporation, accompanied by the decrease in the survival rate and maturation of DCs.

Change Character of Electric Conductive Properties of Weeds Plants Tissue at Electric Impulse Damage

Currently, in order to obtain pre-planned results and increase the efficiency of using existing technologies, electrotechnological operations are applied in the processes of agricultural production, food and processing industries, including those that implement the effects of high voltage impulses. One of such promising technologies in agriculture can be considered to be electric impulse weeding that is the fight against unwanted and weed vegetation. Such operation, which is carried out with high rates of technological efficiency, is environmentally friendly and refers to high-tech processes. For practical applications and technical implementations, it is necessary to study various properties and parameters of effected objects – weeds. The partial dependence of the electrical resistance of the plant tissue or the dispersion of resistance over the frequency of the measuring current was decided to be used as the studied characteristic. This approach allows to consider weeds not only as a load with which the electrical energy supply system is in contact but also to thoroughly study how this resistance changes during processing. The conducted experimental studies made it possible to clarify the nature of the change in resistance under the damage process to plant tissue, as well as to estimate the quantitative indicators of both the energy absorbed by the plant tissue and the ranges of variations of its resistance. The change character in the plant tissue resistance at the process of the electrical impulse damage remains unchanged, only the resistance value at low frequencies decreases. The total damage corresponds to the period of processing when the resistance of the plant tissue is practically equal at low and high frequencies. This behavior of the mentioned parameter is typical for internal and surface tissues of weeds, as well as for their root systems and leaf-stem parts. Theoretical verification of the change character in the plant tissue resistance of destroyed weeds confirmed the description of the proposed mechanism of change in resistance in the process of irreversible electrical impulse damage.

Energy/information dissipation and blood flow in human body

The amount of work done to displace blood in systemic arteries and capillaries exceeds the work done by the left ventricle. Besides, at the heartbeat, electromagnetic energy dissipates from the heart to the whole human body. For the problem study, the dielectric spectroscopy method was used. Ringer’s, amino acid solution, and heparinized venous blood were affected by the external electromagnetic oscillations (100-65000Hz, 1-8MHz.) in 17 healthy individuals. Correlations were noted between the initial and induced signal forms/frequencies according to the impedance of the system. The electric impulse from the heart initiates an oscillating electric field around the charged cells/particles and an emerging repulsing electromagnetic force, based on the electroacoustic phenomena, promotes the blood flow, in addition to the pulse pressure from the myocardial contraction. Blood conduces mechanical, electromagnetic waves of different frequencies and transmits energy/information to implement the spontaneous chemical processes in the human body.

Strain improvement of native Saccharomyces cerevisiae LN ITCC 8246 strain through protoplast fusion to enhance its xylose uptake

Co-utilization of xylose and glucose and subsequent fermentation using Saccharomyces cerevisiae could enhance ethanol productivity. Directed engineering approaches have met with limited success due to interconnectivity of xylose metabolism with other intrinsic, hidden pathways. Therefore, random approaches like protoplast fusion were used to reprogram unidentified mechanisms. Saccharomyces cerevisiae LN, the best hexose fermenter, was fused with xylose fermenting Pichia stipitis NCIM 3498. Protoplasts prepared using glucanex were fused under electric impulse and fusants were selected using 10% ethanol and cycloheximide (50 ppm) markers. Two fusants, 1a.23 and 1a.30 showing fast growth on xylose and tolerance to 10% ethanol, were selected. Higher extracellular protein expression observed in fusants as compared to parents was corroborated by higher number of bands resolved by twodimensional analysis. Overexpression of XYL1, XYL2, XKS and XUT4 in fusants as compared to S. cerevisiae LN as observed by RT-PCR analysis was substantiated by higher specific activities of XR, XDH and XKS enzymes in fusants. During lignocellulosic hydrolysate fermentation, fusants could utilize glucose faster than the parent P. stipitis NCIM 3498 and xylose consumption in fusants was higher than S. cerevisiae LN.

Towards programmable friction: control of lubrication with ionic liquid mixtures by automated electrical regulation

For mechanical systems in relative motion it would be fascinating if a non-mechanical stimulus could be used to directly control friction conditions. Therefore, different combinations of lubricants and external triggers for tribological influence have already been investigated. We show that when two metallic friction partners are lubricated with ionic liquid mixtures (ILM), consisting of long-chain cation and two different high charge/mass ratio anion containing ILs, the application of an electric impulse induces a permanent change of the frictional response. Such mixtures are able to alter the coefficient of friction (COF) to a greater extent, more accurately and faster than the respective single-component ILs. This change in the frictional properties is presumably due to changes in the externally induced electrical polarization at the surface, which influences the molecular adsorption, the exchange of adsorbed ions and their molecular orientation. The correlation between surface charges and friction can be used to control friction. This is achieved by implementing an electric tribo-controller which can adjust preset friction values over time. Programming friction in this way is a first step towards tribosystems that automatically adapt to changing conditions.

Providing Boot-Type Injection Rate Shape by Electric Impulse Control of the Common Rail Injector

For effective reduction of noise level and nitrogen oxide content in exhaust fumes of diesel engines, multistage fuel injection is used in combination with control of the front edge shape of the main injection. At the Moscow Automobile and Road Construction State Technical University (MADI), a method of control of the injection rate shape using an electric impulse was proposed, which was applied to the electromagnet of the control valve of the injector of the common rail fuel system. A computational and experimental analysis of the possibility of boot-type injection rate shape was carried out. The studies involved three most used designs of the common rail injector (CRI): CRI 1 featuring a control valve with shut-off cone and piston; CRI 2 consisting of a flat-lock control valve and a needle, which does not overlap the drain when the needle is in the highest position; CRI 3 with an injector that partially overlaps the drain. It was established that friction in the control valve piston and the guide surface pair of CRI 1 complicated the implementation of the boot-type injection rate due to its smoothing. CRI 2 and CRI 3 provide boot-type injection rate at different pressures in the fuel accumulator. The CRI 3 example shows that the instability of fuel supply during boot-type injection rate is comparable with that of fuel pre-injection, which is widely used in the organization of the common rail diesel engines working process.