Archive

Recovery of the neural networks after brain injury can be studied in vitro using cell patterning technologies such as microfluidics. In this study, neural tissue recovery of cortical networks was modeled in a three-chamber microfluidic chip by growing two weakly coupled neuronal networks and by plating new immature cells between it. The direction of synaptic connections was formed by the asymmetric design of the microchannels in the chip. We investigated dynamics of morphological characteristics of neurites growth through the microchannels and spiking activity propagation between the networks after integration of new cortical cells. The results can be used for the development of new approaches for brain functional recovery after injury.

In the last decade, glial cells, primarily astrocytes, have received increasing attention in studying various aspects of brain functioning. Although astrocytes are not electrically excitable cells, they are able to generate and transmit Ca²⁺ signals, which can propagate from cell to cell, forming «calcium waves». Calcium signaling allows astrocytes to interact with each other, as well as regulate neuronal function. Despite numerous studies on calcium events in astrocytes under various pathological conditions, the features of astrocytic calcium dynamics in aging are currently understudied. Our work aimed to analyze the features of calcium activity in primary astrocyte cultures during physiological and accelerated aging, as well as to assess the effect of hypoxic damage on calcium activity. For this purpose, we developed an algorithm for the accurate detection of calcium events and the calculation of indicators describing the main parameters of calcium events in a cell. It was shown that the frequency of calcium event generation in astrocytes decreases during aging while their duration and amplitude increase. Hypoxia aggravates the changes caused by cell aging.

Erythrocytes, performing their basic oxygen transport function, simultaneously affect blood viscosity by changing the deformability of their structure, and also have a vasodilating effect on the walls of blood vessels using NO. An important role in this regulation is played by the purinergic signaling system, which was confirmed by this study of the morphofunctional parameters of red blood cells in the presence of ATP and sodium nitroprusside (SNP). We found that under the action of 0.5 mM ATP on red blood cells, the ability of hemoglobin (Hb) to bind oxygen decreases against the background of a slight increase in complexes with oxygen, while the total number of membrane-cytoskeletal proteins also decreases. This, in turn, is accompanied by a redistribution of Hb molecules and an increase in the area of ​​red blood cells. In the presence of 5.0 mM ATP, the oxygen transport function of erythrocytes and the quantitative composition of membrane proteins change similarly, while the geometric height and volume of the cells are significantly reduced. The combined effect of 100 μM SNP and 0.5 mm ATP has the greatest effect on the conformation of hemoporphyrin molecules, which leads to a sharp increase in hemoglobin complexes with oxygen, while the affinity of hemoglobin for oxygen decreases. The membrane-cytoplasmic component of the cell also undergoes changes. Thus, ATP, both separately and in combination with SNP, affects the oxygen transport and regulatory function of red blood cells, activating the purinergic signaling pathway and triggering a cascade of adaptation reactions in the cell.

Along with the wide spread of bacterial antibiotic resistance over the world, the treatment efficiency of infectious disease is greatly affected by the mixed biofilm formation by pathogenic bacteria. Staphylococcus aureus and Pseudomonas aeruginosa, a frequent cause of nosocomial infections, exhibit both synergistic and antagonistic interactions in co-culture, leading to various changes in the metabolic profile of bacteria, which in turn affect their sensitivity to antimicrobials. Here we show that S. aureus cell-free culture liquid exhibits bacteriostatic properties and increases the efficacy of antimicrobials against P. aeruginosa. Thus, the MICs of amikacin, gentamicin, and ciprofloxacin decreased 2-4 fold in the presence of cell-free supernatant of S. aureus 24 h culture. Furthermore, the combination of the latter with antimicrobials increased the efficacy of amikacin up to 64-fold. Thus, the combined use of cell-free culture liquid of S. aureus with broad-spectrum antibiotics can be used to increase the effectiveness of antimicrobial therapy of P. aeruginosa.

Microbial production of testosterone from phytosterol is a promising alternative to the chemical synthesis from androstenedione, which is currently used. It is possible using wild-type or recombinant strains of Mycolicibacterium neoaurum. One of the key steps of biotechnology development is inoculum preparation which determines the volumetric/mass ratios of the bioreactors and affects the biotransformation rate and duration. In the present work, the conditions for the growth of testosterone-producing strain of Mycolicibacterium neoaurum recombinant strain with enhanced 17β-HSD activity were optimized. The effects of medium composition, temperature, and glucose supplements on biomass size and density were estimated. The results are of importance for the development of microbial technology for testosterone production.

In this paper, we present the data on the properties of the probiotic strain Bacillus intestinalis GM2 related to adhesion, auto-, and coaggregation. GM2 exhibits a strong autoaggregation phenotype. The autoaggregation ability of GM2 did not change after trypsin treatment but was reduced under the influence of extracellular culture metabolites. Coaggregation of probiotic strains with pathogens is one of the mechanisms of displacement of pathogens from the intestinal microbiota. It was shown that the GM2 strain exhibits the ability to coaggregate with tested strains of Escherichia coli, Enterococcus faecalis, Staphylococcus aureus, and Bacillus cereus. The studied properties of bacilli can be useful for the creation of new probiotics for poultry farming. 

Nowadays, manned cosmonautics is faced with the task of carrying out a long-term space flight beyond the limits of low Earth orbit. Under the conditions of an orbital space flight, a person is exposed to a number of adverse effects on the body, among which microgravity is especially distinguished. Prolonged exposure to microgravity can lead to severe immune impairment. At present, immunological studies of cosmonauts can be conducted only after they return to Earth at the end of a space flight, as a result of which the use of ground-based models that imitate specific space factors seems to be an advanced direction. A well-studied microgravity model is bed rest, during which volunteers are kept in strict rest in bed. Granulocytes, as representatives of innate immunity, are the first among immune cells to respond to an altered state of the body; therefore, researches of the influence of bed rest on the granulocyte phenotypic characteristics can provide important information for the development of prophylaxis measures to the immune disorders’ development when exposed to microgravity. The work used data obtained from six subjects. The impact of bed rest was determined at the end of the model, after 21 days. The following granulocyte clusters of differentiation (CD) were studied by flow cytometry: CD25, CD64, CD23, CD14, CD16, CD36, CD11b, CD18, CD286. Tendencies to a decrease in the percentage of CD64⁺ subpopulation and an increase in the percentage of CD23⁺, as well as CD25⁺ subpopulation of granulocytes after 21 days of bed rest were revealed.

Radiotherapy is one of the most effective and most commonly used methods of cancer treatment. However, as a result of irradiation, there are side effects that occur as a result of ionizing radiation on healthy tissues. The use of a combined approach with the use of low doses of radiation and antitumor drugs that have a radiosensitizing effect may be one of the ways to reduce side effects and overcome the resistance of malignant cells. This study was undertaken to evaluate the combined effects of radiotherapy and the chemotherapeutic agent Doxorubicin on A431 human epidermoid carcinoma cell line. The cells were incubated with the antitumor antibiotic Doxorubicin and then exposed to high-energy electron ionizing radiation. The cell viability was examined using the MTT assay. The results showed that Doxorubicin acts as a radiosensitizer. Moreover, the combined effect of Doxorubicin and high-energy ionizing radiation of electrons is additive. According to the obtained results, combination therapy used in the treatment of oncological diseases can significantly reduce the radiation dose and minimize the side effects that occur during high doses of irradiation.

Aim of the study: to analyze the results of performing retrograde perfusion of the pulmonary artery during an open thromboembolectomy from the pulmonary artery. Materials and methods: the experience of performing retrograde perfusion of the pulmonary artery in 10 patients operated in our clinic for massive pulmonary embolism is presented. Retrograde perfusion was performed after the stage of embolectomy from the pulmonary artery. For the latter, we used a disposable cardioplegic solution delivery system and 2 cardioplegic pumps of the heart-lung machine: the first for taking blood perfusate from the oxygenator, the second for supplying the combined solution. The blood perfusate and solution were mixed in a 3:1 ratio and injected selectively into the orifices of the pulmonary veins under a pressure of up to 20 mm Hg. (volume perfusion rate 200-250 ml/min) for 4 minutes. Results: despite the initial severity of the patients' condition, as well as the amount of surgical intervention performed, the hospital survival rate was 100%. Along with this, we did not note the development of specific complications, as well as the aggravation of the course of the intraoperative and early postoperative periods. Conclusion: retrograde pulmonary artery perfusion is a very encouraging and promising technique that provides effective and safe removal of small thromboembolism from the peripheral parts of the pulmonary arterial bed, as well as preventing the development of residual pulmonary hypertension as a result of developing intraoperative air embolism.

Viliuisk encephalomyelitis (VE) is a neurodegenerative disorder that afflicts the aboriginal people of Yakutia in Siberia. The disease is characterized by a progressive duration and aseptic inflammatory episodes, with intrathecal synthesis of oligoclonal IgG (OCBs) in some patients. The aim of this study was to evaluate the role of soluble ligands and receptors of the tumour necrosis factor (TNF) superfamily as potential participants in VE pathogenesis. To achieve this goal, we measured the levels of sTNF-α, sFas-L, sTRAIL, sCD40L ligands, and sCD40 receptor by ELISA in the plasma of VE patients compared with healthy individuals of the same population and patients with demyelinating diseases, including multiple sclerosis (MS) and neuromyelitis optica (NMO), as examples of disorders involving immune pathology. In addition, the same markers were analyzed in the CSF of VE patients and patients with demyelinating diseases. The results obtained showed that the increased level of plasma sTNF-α in VE patients was associated with the detection of OCBs (p = 0.01; two-tailed Student’s t-test). The sCD40L level in plasma was significantly increased in VE patients, regardless of the presence of an inflammatory component (p = 0.001; Student's t-test), and their healthy relatives (p = 0.004; Student's t-test). Our results suggested that increased blood sCD40L levels are associated with the chronic form of VE and may participate in the predisposition to the disease. Increased blood sCD40L levels may lead to pathology of the vascular endothelium in the brain and the development of VE pathology.

Prenatal hypoxia remains the leading cause of infant mortality and severe disability in newborns. Disturbances in the development of fetal brain structures and functions due to hypoxic damage are the main trigger for the development of severe neurological disorders and accelerated neurodegeneration processes and can also be the cause of epileptiform activity in the postnatal period. Herein, the role of chronic prenatal hypoxia on the functional state of C3H+C57Bl6 hybrid mice during the first three weeks of postnatal development and the risks of developing epileptiform activity when provoking audiogenic seizures were assessed. Exposure to chronic prenatal hypoxia was found to increase the risk of neonatal mortality and developmental delay in the surviving individuals in the first two weeks of the postnatal period. It was shown that one of the causes of the failure of adaptation might be the disruption of the functional activity of the mitochondrial apparatus of brain cells mediated by the disruption of the first and second respiratory chain complexes. Exposure to chronic prenatal hypoxia has no significant effect on the increased risk of seizure activity in mice during audiogenic stimulation in late postnatal development, does not activate the development of persistent neurological deficit, and does not significantly affect the cognitive functions and learning ability of animals.

The study of mitochondrial DNA (mtDNA), particularly the hypervariable segment (HVS1) region, is widely used to reconstruct a population's history, structure, and origin. The origin of the Sakha people living in the north-east of Russia has been discussed for more than 300 years, but up to the present time many aspects of their ethnogenesis remain unclear. In order to clarify the migration routes of the ancient ancestors of the Sakha, we analyzed the mitotypes of 69 unrelated representatives of this ethnic group, whose belonging to the ethnic group was traced to the third generation. In the studied Sakha group, we identified 33 mitotypes, the distribution of which by haplogroups approximately coincides with the data of other studies. The results of a comparative study of Sakha mitotypes according to EMPOP data and literature sources revealed a wide distribution of the identified mtDNA HVS1 haplotypes in many populations of Eurasia. A comparison of the obtained mitotypes with the results of mtDNA sequencing of ancient samples shows that most of the mtDNA lines of modern Sakha have long been located on the territory of Yakutia. West Eurasian and East Asian mtDNA lines were incorporated into the Sakha genome at different times in different ways. The results obtained contribute to a better understanding of the routes of ancient migrations of the ancestors of the Sakha population.

The aim of the work was established as to study the effect of low-intensity laser therapy (LILT) on the morphology of neutrophils by the method of interference microscopy in correlation with metabolic parameters in the use of modulators which permits to assess the contribution of various structures of neutrophils to the realization of LILT effect. There was in vitro research. The neutrophils of rats served as the object of research. The neutrophils were incubated with adrenalin, cortisol, and β-blockers of adrenoreceptors. The effect of LILT in combination with a preliminary incubation of these substances was studied. Autonomous laser shower MarsiK (R&D company “Petrolaizer,” Saint Petersbourg), a laser therapeutic complex for animals, was used as a laser irradiation source. The wavelength of the irradiation was 830 nm, and the total power was 90 MW. The phasometry of neutrophils was studied by the laser modulation interference microscopy method, the processes of lipid peroxidation were analyzed by the malondialdehyde concentration, and the ATP concentration was defined by spectrophotometry. LILT provoked unidirectional changes in indices in the groups where the neutrophils were incubated with adrenalin, cortisol, and adrenoreceptor blockers. It manifested in the intensification of lipid peroxidation, increase in ATP concentration, and on the other hand, an increase in phase diameter of neutrophils regarding indicators without the impact of LILT. It is shown that the LILT effect intensity may be modified depending on stress realizing hormones (adrenalin and cortisol), β-blocker of adrenoreceptors. It permits us to suppose that the LILT effects may be realized in different ways depending on the state of the cells.

Cisplatin, as an antineoplastic drug belonging to the platinum family, has severely nephrotoxic and neurotoxic side effects. L-carnitine (LC) is an antioxidant-rich natural substance. The notion that LC may play a protective function in Cisplatin-induced nephropathy and neuropathy was investigated in this study. Nephropathy was created by a single intraperitoneal injection of Cisplatin at 20 mg/kg body weight, while neuropathy was induced by daily intraperitoneal injections of Cisplatin at 2.3 mg/kg body weight over two rounds of five days, with five days break in between. The rats were subsequently given LC at a dose of 250 mg/kg body weight, followed by estimation of serum urea and creatinine with histological examination of renal tissue for the nephropathy group and evaluation of nerve conduction velocity for the neuropathy group. In comparison to the nephropathy group, blood urea and creatinine levels were significantly lower after treatment with LC. Furthermore, LC therapy improved the physiological characteristics of the sciatic nerve significantly. In conclusion, the significant impairment of renal function and the decrease in sciatic nerve conduction velocity induced by Cisplatin may be avoided if L-carnitine is administered as a preventative medication.

The search for new molecular targets to protect brain cells from ischemic damage has remained one of the most urgent tasks of neurobiology and medicine over the past decades. The modern concept of the glucose-lactate shuttle, the main mechanism for providing energy support to neurons under conditions of increased physiological activity, implies a functional fusion of neuron and astrocyte metabolism, which becomes particularly important under energy or oxygen starvation conditions. The transfer of energy substrates in response to increased glutamate release by the presynaptic terminal occurs through monocarboxylate transporters (MCTs) and depends on a large number of endogenous mechanisms of homeostasis. Our work examined the role of MCTs in the implementation of the neuroprotective effect of glial cell-derived neurotrophic factor (GDNF), one of the key participants in the regulatory system capable of maintaining the viability and functional activity of neurons under conditions of energy starvation. It has been shown that the application of the MCT1 transporter inhibitor, even under normal conditions, significantly affects the parameters of spontaneous bioelectrical activity of neural networks and cell viability of primary dissociated hippocampal cultures. However, the authors’ data on the role of this type of transporters in glucose deprivation and the effect of MCT blockade on the neuroprotective effects of GDNF are of the greatest interest.

In this study, we investigated the toxic properties of synthesized immunomagnetoliposomes of the following composition: phosphatidylcholine - cholesterol - disteroylphosphoethanolamine-polyethylene glycol (2000) - magnetite - antibodies to histone H3. Their dimensional characteristics were studied by the dynamic light scattering method. It was revealed that liposomal nanoparticles have dimensions of 176.4 ± 12.9 nm. Data on the toxicity of the synthesized nanoparticles in relation to human blood cells: liposomes in cells to liposomes ratio from 1/1 to 1/1000 have no toxic effect on lymphocytes; lactate dehydrogenase (LDH) release from human blood erythrocytes has been observed in the case of cells/liposomes ratio 1/100-1/1000 after the incubation with liposomes containing sodium azide (0.03%).

Purpose of the study: substantiation of indications and assessment of results X-ray endovascular treatment of "critical" PE in patients with acute hemorrhagic cerebrovascular accident. Material and methods: the study included 47 patients with acute violation of cerebral circulation for hemorrhagic and mixed types complicated by massive pulmonary thromboembolism of high risk, with "critical" manifestations of right ventricular failure. Depending on the method of PE treatment used, the patients were divided into 2 groups: the study group (17 people), in which endovascular mechanical fragmentation of thromboembols was performed, and the control group (30 people), in which only basic intensive therapy was used. Results: patients of the study group underwent thromboembolic fragmentation in order to transfer embolism of the trunk and the main branches into embolism of smaller branches of the pulmonary artery. Technical success of the procedure (destruction of the central thromboembolus) was achieved in 100% of the cases. 14 patients (82.4%) showed positive clinical dynamics in the form of an improvement in the general condition, a decrease in pressure in the pulmonary artery, and a decrease in the volume of the lesion of the pulmonary bed according to CT angiography. Three patients (17.6%) died in the early postoperative period. In the control group of 30 patients, 25 patients died, hospital mortality was 83.3%. Conclusion: the method of catheter endovascular fragmentation of thrombi for the treatment of "critical" pulmonary embolism in patients with acute hemorrhagic cerebrovascular accident leads to a rapid and safe decrease in pulmonary artery pressure. 

The actuality of the problem of thermal trauma is determined by its high prevalence. Severe disorders of central, regional and peripheral hemodynamics are developing with a predominant violation of microcirculation and metabolic processes in the body during burn disease. Promising NO donors suitable for biomedical use are dinitrosyl iron complexes (DNIC). The aim of the study was to evaluate the effect of DNIC on the state of microcirculation in normal conditions and on the model of combined thermal trauma (CTT). The experiments were carried out on white male rats of the Wistar line. CTT (contact burn on the area of 20% of the body surface and thermal inhalation exposure to hot air and combustion products for 20-30 seconds) was applied under anesthesia. Animals with CTT were treated daily with intraperitoneal injections of a 10% DNIC solution (1 ml; 0.3 micromol/l). Laser Doppler flowmetry was used to assess the dynamics of the microcirculation state. The physiological and biochemical mechanisms of microcirculation disorders in CTT are revealed in the work. There was an increase in the endothelial, neurogenic and respiratory components of microcirculation, microhemodynamics of the border area of the burn, and the index of the microcirculation bypass. It was shown that when using 0.3 micromol/l of DNIC in rats under normal conditions, there is a decrease in the neurogenic and respiratory components, an increase in perfusion and the myogenic component of the microcirculatory channel. The role of 0.3 micromol/l of DNIC in the normalization of microcirculation in CTT was established.

The work aimed to study the protein spectrum, morpho-functional state of erythrocyte membranes under stress and correction by low-intensity laser radiation. The experiments were carried out in vitro. Low-intensity laser radiation with a wavelength of 830 nm and a power of 90 mw was used for irradiation. Laser therapy was performed using an autonomous laser shower "MarsIK" (NPO "Petrolaser", St. Petersburg). The protein fractions of red blood cells were analyzed by electrophoresis, the morphology of red blood cells by laser interference microscopy, the еletrophoretic mobility of erythrocytes was measured by microelectrophoresis in our modification and the concentration of malondialdehyde in red blood cells by reaction with thiobarbituric acid spectrophotometrically. It is proved that the change of protein fractions in the erythrocyte membrane under stress, a decrease in the amount of spectrin, band 3 protein, glycophorin and ankirin. The stress is characterized by an increase in the number of echinocytes, stomatocytes and degenerative-altered red blood cells. Exposure to LLLT on blood samples determined the recovery of the studied parameters to the control group values (physiological norm). The role of the discovered metabolic changes in RBC in stress and their correction by LLLT is under discussion.

Increase in the life expectancy of the population is associated with a tendency of aging in patients with chronic heart failure (CHF). Gender, age and genetic differences of the individual risk factors are still discussed in the aspect of influence on the heart failure clinical phenotype. The study aimed to identify the gender and age differences of chronic heart failure clinical characteristics in terms of the ZBTB17 gene (rs10927875) polymorphism. Materials and methods. A total of 351 patients with CHF of ischemic etiology of both genders were examined. The average age of men was 65.3±9.9, women  ̶  69.7±9.61 years. Results and conclusions: CHF of ischemic etiology in the absence of differences in the burdened family history, previous myocardial infarction (MI) observed mainly in middle-aged and elderly men and elderly and senile women. Significant gender differences in comorbidities were revealed: in 40% of men, CHF was associated with chronic obstructive pulmonary disease (COPD), in 47.3% of women ‒ with chronic kidney disease (CKD) and in 32.2% - with diabetes mellitus (DM). The distribution of rs10927875 polymorphism ZBTB17 genotypes were corresponded to the Hardy-Weinberg equilibrium (χ2 = 0.272, p = 0.873). CC genotype in men was associated with reduced left ventricular ejection fraction (LVEF). CT genotype in women associated with CHF FC III and IV due to the frequent indications in the past on myocardial infarction, concomitant DM, obesity and hypercholesterolemia. The CT genotype rs10927875 was characterized by a frequent combination of CHF with COPD in men and CKD and DM in women.

The aim of the work was to study the effects of photobiomodulation (PBM) in the red spectrum (640 nm) with fluences from 3 mJ/cm² to 2 J/cm² in combination with ionizing radiation (IR) at doses of 2-6 Gy against human BJ-5ta-hTERT - postnatal fibroblasts. The cells were exposed to low-intensity red light before or after their exposure to IR, the viability of the cells was determined by MTT 24 hours after the last exposure. It is shown that the effects of PBM depend on the fluence of PBM, the dose of IR and the sequence of the actions of these physical factors on cells. The adaptive effect of PBM was observed only for high fluences-1 and 2 J/cm² when exposed to PBM and subsequent (after 1 hour) irradiation of IR. At the same time, the stimulating effect of PBM was observed only for low fluences from 3 to 300 mJ/cm² under IR irradiation and subsequent (after 1 hour) exposure to PBM. These data should be taken into account when using PBM for the correction of adverse events of radiation therapy in an oncological clinic.