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Introduction. Spinal cord injury is a debilitating traumatic event in central nervous system resulting in tissue destruction and severe neurological deficit development. Preclinical assessment of quantitative lesion area parameters (e. g. structure and volume) is critical for subsequent evaluation of neuroprotective and / or neuroregenerative therapy efficiency. Current methods for parameter calculation require manual limitation of the interested area (region of interest, RoI). This process is tedious and often not precise enough.Study objective is to develop and implement software for automated assessment of volume and structure of posttraumatic spinal cord lesion using extra-high-field MRI 7.0 Tesla and to compare methods preciseness with the current manual techniques.Study design. Ten rat models of acute severe spinal cord contusion injury were used including female Sprague–Dawley animals weighting 250–350 gr. MRI imaging was performed in 1 day postoperative and then 4 times with interval (1 week). Study was prospective open-label uncontrolled comparative.Materials and methods. Standard spinal cord contusion injury model was used. Anesthetized animals underwent laminectomy at level Th9–Th10 vertebrae followed by “weight drop” injury technique application: 10 g weight with 2 mm pin diameter dropped from 25 mm height. Software was developed using Microsoft Visual Studio 2017 environment and programming language C#. Statistical analysis was performed using IBM SPSS Statistics 21.0 software.Results. We developed and patented specialized software Spinal cavity Searcher realizing the algorithm of T2‑weighted images (T2‑WI) analysis based on image bynarization and Freeman chain code. This algorithm supports calculation of spinal cord posttraumatic lesion parameters in a half-automatic manner. Results of this algorithm application were comparable to results of manual calculation: no statistical difference were observed between two values.Conclusion. Current method of spinal cord injury volume and structure quantitative assessment simplifies the calculation procedure due to automatization of RoI limitation comparing to manual technique. The level of preciseness is comparable in both methods.Clinical relevance. The developed algorithm optimizes the process of non-invasive control of the performed treatment efficiency according to 7.0 Tesla MRI data.

Transplantation of various types of stem cells as a possible therapy for stroke has been tested for years and the results are promising. Recently, most researchers are inclined to assume that the therapeutic effect of stem cell therapy is based on the mechanism of paracrine action associated with the secretion wide set of regulatory proteins. The aim of this study was to evaluate therapeutic effects of iPSC-derived glial progenitor cells conditioned medium in the rat middle cerebral artery occlusion model of the ischemic stroke. We showed that intra-arterial administration of glial progenitor cells conditioned medium promoted faster decrease of neurological deficit compared to the control group. Moreover, expression of gap43, bax, and tnfa genes involved in neuritogenesis, apoptosis and neuroinflammation was altered. However, no significant enhanced reduction of the infarct volume was registered. Our results demonstrated that administration of glial progenitor cells conditioned medium induced functional recovery after experimental stroke and may affect brain plasticity.

We developed a viral vector Ad5/35-CAG-mBDNF expressing the mature form of BDNF (mBDNF). On the basis of olfactory ensheathing cells transduced with this adenovector, a new gene-cell construct was obtained. In experiments in vitro, high viability of the transduced olfactory ensheathing cells and enhanced secretion of BDNF by these cells were observed. It is possible that a new gene-cell construct will significantly increase the regenerative effects of transplanted olfactory ensheathing cells.

A gene-cell construct based on rat olfactory mucosa ensheathing cells transduced with an adenoviral vector encoding a mature form of brain neurotrophic factor (mBDNF) was transplanted into post-traumatic cysts of rat spinal cord. Transplantation of the gene-cell construct improved motor activity of the hind limbs and reduced the size of cysts in some animals. However, comparison of the effects of transduced and non-transduced ensheathing cells revealed no significant differences. In parallel in vitro experiments, a decrease in the proliferation of transduced cells compared to non-transduced cells was observed. It is likely that mBDNF reduces proliferation of transduced cells, which can affect their efficiency. The therapeutic efficacy of the new gene-cell construct is most likely provided by the cellular component.

Neurotrophin-3 enhances the effectiveness of human olfactory ensheathing cells in improving hind limb mobility in rats with post-traumatic cysts of the spinal cord. Transplantation of olfactory ensheathing cells into spinal cord cysts reduced their size; neurotrophin-3 did not modulate this effect. Combined preparation of human olfactory ensheathing cells and neurotrophin- 3 can be used in neurosurgery for the treatment of patients with spinal cord injuries.

Visualization of transplanted stem cells in the brain is an important issue in the study of the mechanisms of their therapeutic action. MRI allowing visualization of single transplanted cells previously labeled with superparamagnetic iron oxide particles is among the most informative methods of non-invasive intravital imaging. Verification of MRI data using pathomorphological examination at the microscopic level helps to avoid errors in data interpretation. However, making serial sections of the whole brain and searching for transplanted cells under the microscope is laborious and time-consuming. We have developed a method for 3D modeling of the distribution of transplanted cells in the brain allowing navigating through various brain structures and identifying the areas of accumulation of transplanted cells, which significantly increases the efficiency and reduces the time of histological examination.

We studied the effect of transplantation of ensheathing cells obtained from the olfactory mucosa of rats and humans on the size of posttraumatic spinal cord cysts. MRI examination showed that transplantation of these cells into experimental posttraumatic cysts of the spinal cord led to a significant decrease in cyst volume and even their complete disappearance in two animals receiving transplantation of rat or human cells. These findings attested to regenerative processes developing as a result of ensheathing cell transplantation. Further studies in this field will be aimed at elucidation of the mechanisms underlying spinal cord regeneration in the area of posttraumatic cysts after transplantation of ensheathing cells.

Depression is a serious mental disorder that affects more than 300 million people worldwide. Due to the lack of effective treatment methods, the pathogenesis of depression is necessary to study in order to understand its development and find new therapies. The review describes the main mechanisms of depression, including the monoamine hypothesis, impairment of the hipotalamic-pituitary-adrenal axis, decreased production of neurotropic factors, and neuroinflammation. Genetic correlations, gene polymorphisms, and epigenetic mechanisms are also considered. Common and different features of the etiology are analyzed for depression and depressive conditions associated with other pathologies (schizophrenia, Parkinson disease, and Alzheimer's disease). Modern experimental methods used to investigate the molecular mechanisms of depressive conditions are described with a focus on gene knockouts in laboratory animals and the CRISPR/Cas technology. Consideration is given to optogenetic and chemogenetic methods and analyses of genetic polymorphisms and their combinations. The data may provide for a better integral understanding of the modern ideas about the pathogenesis of depression as an isolated or comorbid disorder and the prospects in studying the mechanisms of depressive conditions.

Оpening Speech of the chairman of the priority council, vice-president of the RAS, academician V.P. Chekhonin

To study demyelination and remyelination processes and their response to different drugs, a protocol for modeling multiple sclerosis using the copper chelator cuprizone was developed. Magnetic resonance imaging confirmed the presence of demyelination lesions on week 4 of 0.6% cuprizone-containing diet. Immunohistochemical staining with polyclonal antibodies to glial fibrillary acidic protein (pAb GFAP) confirmed the increase in the number of reactive astrocytes on week 4 of diet and during remyelination (week 2 after diet). Analysis of neurophysiological functions in mice with cuprizone-induced demyelination revealed motor and behavioral deficits. This model can be used as a tool for preclinical studies of the efficiency of multiple sclerosis diagnostic and therapy.

The opportunities and the most promising ways of using cellular technology in traumatic spinal cord injury are considered in this review. A large number of experimental and clinical studies with the use of different types of cells: embryonic stem cells, induced pluripotent stem cells, mesenchymal stem cells, Schwann cells, olfactory mucosa cells, and others – was conducted. The use of these types of cells in traumatic spinal cord injury treatment often demonstrated a positive therapeutic effect: the motor and sensory function recovery of the spinal cord. However, some types of cell preparations involve some methodological and ethical problems; some types of cell therapies are ineffective or give rise to side effects. These factors complicate the selection of optimal cell therapy for the traumatic spinal cord injury treatment. The most promising cells seem to be the cells of the olfactory mucosa. Getting the olfactory mucosa is considered to be a feasible and safe procedure for patients. The clinical application of the cells of the olfactory mucosa is effective in motor function recovery due to remyelination and axonal regeneration after spinal cord injury. These cells are tissue-specific and autologous since they can be obtained from a patient with spinal cord injury, and after cultivation, expansion, and directed differentiation they can be transplanted to the same patient. The presented benefits of olfactory mucosa cells open up the possibility for its clinical application in the cell therapy.

The review is devoted to actual problems of studying liposomes and other lipid nanoparticles that, being loaded with active substances, can be widely used in biomedical applications.

Glioblastoma (GBM) is the most common type of primary brain cancer that characterized by poor prognosis due to the rapid progression, active angiogenesis, enhanced tumor cell invasion and the emergence of resistance toward conventional therapy. In this connection, nowadays, new approaches for selective inhibition of crucial steps in tumor progression are actively developing. The key feature of tumor growth and development is angiogenesis. VEGF and its receptor VEGFR2 play the pivotal role in regulation of tumor vessel formation. Therefore, VEGFR2, as the main receptor of VEGF's pro-angiogenic signal transducer, is a promising molecular target for anti-angiogenic therapy. There is evidence that inhibitors of VEGF and VEGFR2 reduce endothelial cell proliferation, migration and survival that lead to regression of vessel density and decrease vascular permeability, thereby slowing tumor growth. Currently, a number of VEGFR2 inhibitors are under clinical trials (ramucirumab, cediranib) and several were approved (sunitinib, sorafenib). Despite the promising results of preclinical studies, the efficacy of antiangiogenic drugs in the clinical practice is significantly lower, mainly, due to rapid adaptation of malignant cells that consists of alternative pro-angiogenic pathways activation, recruitment of endothelial progenitor cells from bone marrow and increasing of the invasive growth. Given the diversity of pro-angiogenic mechanisms, enhancement of the efficacy of tumor therapy could be achieved by specific inhibition of VEGFR2 functions that will be supplemented by other antiangiogenic drugs (anti-VEGF,-PIGF,-HIF1alpha). In addition, multitargeting therapy should focus on the combined inhibition of angiogenesis, invasion, metastasis, proliferation and survival of tumor cells.

A review describes general trends and directions of personalized medicine. These include, but not limited to, prediction of disease based on genomic data, diagnostics and therapy monitoring using genomic and postgenomic technologies as well as therapeutic drug monitoring and regenerative cell technologies. The personalized medicine is considered in terms of results of the Human Genome Project and succeeding Human Proteome Project. An importance of personalized approach for modern medicine is emphasized.

Solid tumor progression largely depends on vascularization and angiogenesis in the malignant tissue. The most prominent among all proangiogenic factors is vascular endothelium growth factor (VEGF). VEGF suppression leads to retrogression of neoplastic vessels and tumor growth restriction. Clinical trials of complex antiangiogenic and chemical therapy of different neoplastic tumors have shown promising results. Nowadays bevacizumab is widely used in breast cancer, colorectal cancer and II-IV stage of malignancy gliomas treatment. Unfortunately, in the majority of cases antiangiogenic treatment led not to full recovery, but only to tumor development restriction. Resistance mechanisms include potentiating of alternative proangiogenic signaling pathways and activation of malignant cell invasive population.

Ischemic stroke was modeled in white pedigreeless rats by the superficial blood vessel devascularization in the sensorimotor cortex. The preparations of neural progenitors--rat embryonic neural stem cells (rENSC) and human olfactory epithelium-derived neural stem cells (hOENSC) and differentiated fibroblasts ("cell control") were transplanted at the perimeter of the devascularized region. These cells marked with vital tracer stayed alive in the brain parenchyma for at least 16 days. The monitoring of contralateral forepaw motor deficit during 8 weeks demonstrated that only rats with rENSC transplantation had the stable and significant improvement of performance in cylinder test and swimming test (forepaw inhibition test) in comparison to "cell controls" and rats without cell transplantation. The maximal difference in the relative values (the efficacy) was 25% to the end of the experiment. There was no difference in the indicators of vibrissae-elicited forelimb placing test between experimental groups. The methodological approach used makes it possible to broaden the study of mechanisms of neural stem cells' therapeutic effect in stroke.

We evaluated the effect of "sidedness" of focal stroke induced by pial blood vessel devascularization in sensorimotor cortex of right or left hemisphere on neurological deficit in rats. Animals were preliminary examined for paw preference (hemispheric "dominance") by means of food reaching test. Data of cylinder-test, vibrissae-elicited forelimb placing test and swimming task (forepaw inhibition test) were assessed to monitor functional deficit weekly for 8 weeks after stroke. No difference between rats with right or left cortical damage was observed during the experiment. Rats with ischemia in "dominant" hemisphere performed significantly better than those with "nondominant" hemisphere lesion. The revealed effect of functional brain asymmetry on the neurological rehabilitation after stroke should be considered in design of preclinical investigation of new therapeutic approaches to stroke.

An immuno-enzyme assay of NSE and MBP in the cerebrospinal fluid (CFS) and blood serum of rats during the development of acute experimental autoimmune encephalomyelitis revealed the increase of NSE, a neuronal marker, and MBP, a marker for myelin. The investigation has shown that the increased level of NSE in CSF was prior to the increased concentration of MBP. The changes of these neurospecific proteins (NSP) in the serum were not the same as in the CSF. The analysis of the ratio between NSP concentrations in CSF to serum revealed the initial marked decrease of BBB permeability to MBP but not to NSE.

Perinatal hypoxy-ischemic brain lesions are one of the main causes of mortality and dysfunction of the central nervous system in the neonatal period accounting for high disability rate among survivors. Numerous animal models were proposed to study this problem in ante-, intra-, and neonatal periods of ontogenesis. This paper is devoted to the analysis of the adequacy of these models. The processes of brain development in laboratory animals are considered along with etiopathogenetic factors that can be reproduced on the models of perinatal hypoxy-ischemic lesions in CNS. The available data on such models and their correspondence to known clinical syndromes are summarized. Current trends in the development of new models of hypoxy-ischemic brain lesions are discussed.

The solution to the problem of transportation of high-molecular substances via the hematoencephalic barrier (HEB) is a necessary condition for the development of theoretical and applied aspects of selective transport of biologically active substances (neurotrops, medications) from blood into the brain. In the last decades, views on the possibility of macromolecular transport through intact HEB have changed substantially. Under physiological conditions, translocation of macromolecular substances via HEB is performed with the help of specific molecular transport systems and by endocytosis. The former mechanism prevails both in intensity and the nomenclature of the substances transported. The permeability of the intact (unchanged) HEB for macromolecules exists to an extent necessary for normal CNS functioning. Destruction of dense contacts in the HEB, local retraction and death of endotheliocytes, destruction of the basal membrane take place in pathological processes in the nervous tissue under the influence of inflammatory factors (cytokines, metalloproteinases etc) induced by activated endotheliocytes, T-lymphocytes, macrophages and glial cells; this in fact opens the paracell way for macromolecular components of blood.

The authors of the review discuss the present-day state of and promising approaches to directed delivery of biological agents into the brain. Special attention is drawn to micellar and liposomal transport through the blood-brain barrier (BBB) targeted by immunochemical vectors, such as native or hydrophobized antibodies to specific antigens located at the BBB or in the brain parenchyma.

Appearance of neurospecific proteins (NSP) outside the brain plays a certain pathogenetic role in the development of autosensitization occurring in many kinds of CNS injuries and diseases. Analysis of modern views of cerebrospinal fluid (CSF) exchange allows us to suppose that NSP are eliminated from the brain tissue within CSF, moving from the subarachnoid space into cranial veins, and by lymphatic way, into deep cervical lymph nodes. Elevation of NSP level in CSF indicates an actual neudegenerative process. Serum levels of NSP are determined by the balance between the elimination of NSP from the brain, on the one hand, and their metabolism and the response of the immune system to the appearance of these autoantigenes in the blood stream, on the other. Basing on their own data on the dynamics of NSP (NSE, GFAR, and MBP) concentrations and the proportions of these proteins in CSF and serum (coefficient of elimination) in rats after ischemic, hypoxic, and autoimmune cerebral lesions, the authors offer an algorithm of pathogenetic evaluation, including, on the one part, a conclusion on the presence or absence of a neurodegenerative process, and, on the other, a conclusion on a normal or lowered rate of NSP elimination (metabolism). The results of such an analysis may have a clinical significance in terms of the development of a pathogenetic therapy, including, in every individual case, not only neuroprotectors, but also pharmaceuticals directed towards correction of the functional condition of the immune system.

Myelin oligodendrocyte glycoprotein (MOG) is a myelin-specific protein of the central nervous system (CNS). It is a member of the immunoglobulin superfamily. The contents of this protein in oligodendrogliocyte membrane is very low (approximately 0.1% total proteins). Functions of MOG are still unknown. It is considered that MOG is an autoantigen capable to produce a demyelinating multiple sclerosis-like disease in experimental animals. The structure, membrane topology, putative functions and role in demyelinating diseases are considered in this review.

PEGylated (stealth) immunoliposomes covalently linked to antibodies against human gliofibrillary acidic protein (GFAP) were prepared by coupling the thiolated monoclonal anti-GFAP antibodies, D4, with a maleimide derivative of the phosphatidyl ethanolamine of the liposomal membrane. Depending on the initial protein-to-lipid ratios, the immunoliposomes prepared (with a diameter of about 70 nm) were coupled with 60 to 240 molecules of the antibodies. In vitro experiments with cultures of the embryonic rat brain astrocytes demonstrated a specific binding of these immunoliposomes, which could be inhibited by the preincubation of the cells with free non-coupled D4 but not with non-specific antibodies. Administered intravenously into rats, the immunoliposomes exhibited a kinetic behaviour typical for the PEGylated liposomes: after 24 h, approximately 20% of the doses injected were still present in the blood; the elimination rate constants were 0.05-0.09 h(-1), the elimination half-lives were 8-15 h. With such a systemic longevity, as well as with such a specificity, these immunoliposomes, non-penetrating through intact blood-brain barrier (BBB), should be useful in delivering pharmacological agents to glial brain tumours (which continue to express GFAP) or to other pathological loci in the brain with a partially disintegrated BBB.

Anthracycline antibiotics with significant antitumor activity are widely used for treatment of various oncologic diseases in spite of their poor pharmacokinetics and severe side-effects. To improve the efficacy of treatment of oncologic patients, liposomal formulations of the anthracycline antibiotics, such as Doxil, TLC D-99, and DaunoXome, have been developed. Pharmacokinetic parameters of liposomal doxorubicin and daunorubicin differ markedly from the parameters of their free formulations. Liposomal anthracyclines display a prolonged circulation time, reduced clearance, smaller volume of distribution, and lower toxicity. Doxil and DaunoXome have been licensed for treatment of AIDS-related Kaposi's sarcoma. Entrapment of anthracycline antibiotics into liposomes coupled with monoclonal antibodies enhances their uptake by tumor cells.

Methods of modeling and criteria of evaluating the pathological process in the central nervous system (CNS) as well as modern technologies of provoking the focal ischemia of the brain in experimental animals are under discussion. The results were analyzed comparatively from the viewpoint of efficiency and adequacy of certain models as well as of the clinical specificity of ischemic strikes in man and of set research goals. A literature analysis confirms that the existing arsenal of methodical schemes provides for choosing the most adequate model of focal ischemia of the brain and to ensure a cerebral infarction of a preset scope and localization; it makes possible also an objective evaluation of pathological processes occurring in the cerebral tissues both at the earliest stages of ischemic lesion and during a relatively long time period comparable with rehabilitation time period. Achievements in the sphere of experimental modeling of focal ischemia of the brain pave the way to further promotion of experimental therapy in acute stroke and open up new research priorities; it concerns primarily research of mechanisms timing the neurodegenerative process after ischemic stroke as well as searching-for and testing of means of stroke prevention and of patients' rehabilitation.

Intraperitoneal injection of ketamine in subanesthetic doses to Wistar rats with unilateral occlusion of the middle cerebral artery caused ipsilateral rotation (2-10 rpm), which was recorded in an automatic rotameter. The optimal dose of ketamine was 50 mg/kg. The animals were examined in an automatic rotameter for 40 min. Motor asymmetry persisted for no less than 2 months after surgery. According to the neurological test (Menzies scale) motor asymmetry in animals with focal brain ischemia persisted for no more than 30 days. The degree of ketamine-induced motor asymmetry in intact rats was 0.10 0.03 rpm.

Neurological, locomotor, and behavioral changes in 20 Wistar rats with permanent proximal occlusion of the middle cerebral artery and 18 control sham-operated animals were monitored for 2 months at 10-day intervals. Neurological deficit was maximum immediately after occlusion (3.0 0.6 points), then progressively decreased, but did not completely disappear (0.70 0.06 points on day 60). In control animals neurological status returned to normal on days 10-15. The degree of ketamine-induced rotational asymmetry was 4.0 0.7 rpm over 40 days after surgery and decreased to 2.5 0.6 rpm on day 60. In control rats this parameter only transiently increased to 1.00 0.03 rpm (tdelta=2.5, p0.05). The time of stay on a rotarod and the latency of passive avoidance in rats with focal cerebral ischemia were lower than in control animals throughout the experiment. The results of complex tests can be used in the experimental search for new drugs for the therapy and rehabilitation of stroke patients.

The results of NSE purification procedure, as well as hybridoma technology of anti NSE monoclonal antibodies synthesis are presented. The employment of this procedure yielded highly purified NSE preparation. The immunization of BALB/C mice with NSE preparation led to sensitization of the immunocompetent cells, which could form hybridomes, producing the anti-NSE monoclonal antibodies, after the confluence with myeloma cells Sp 2/0-Ag 14. The ELISA test-system for NSE analysis was developed on the basis of highly purified NSE preparation and monoclonal anti-NSE antibodies. This system was characterized by high specificity, accuracy and reliability. This system may be recommended for analysis of blood-brain barrier functions in the neurological and psychiatric diseases.

This paper reviews recent achievements of the employment of prostate specific membrane antigen in the diagnostics of prostate cancer.

Cell preparations of ventral mesencephalon obtained from 8-, 14-, and 16-17-day rat embryos were stereotactically transplanted to homologous rats with 6-hydroxydopamine-induced hemiparkinsonism. Automated analysis of apomorphine-induced motor asymmetry for 3 months after neurotransplantation revealed higher efficacy of cell preparations from 8- and lower from 16-17-day-old embryos. These data correlated with histomorphological findings, in particular, with the size of grafts, glial reaction, and the number of dopaminergic neurons in the grafts.

The study was made of 17 patients with expected Alzheimer's disease (AD), 29 patients with Parkinson's disease (PD) and 7 with a expected dementia with Levy bodies (DLB). The severity of cognitive disorders was determined according to the following scales: Global Deterioration Rating Scale, Mini-mental State Examination, Mattis Dementia Rating Scale. Besides, the patients state was evaluated, in the whole, according to some scales. Permeability of hemato-encephalic barrier was evaluated according to the blood serum levels of 3 neurospecific proteins--neuron specific enolase, glial fibrillary protein and alpha-glycoprotein. Their determination was performed by ELISA method. The significant elevation of the levels of the proteins studied was found already on the early stages of the disease. Their levels were higher in the patients with the dementia as compared with the individuals without it. There were no differences in cortical, subcortical and combined types of dementia. The authors believe, that some of the proteins studied (neuron specific enolase, for example) may serve as non specific markers of cerebral degeneration.

Methods of GFAP purification and obtaining of hybridoma cells producing monoclonal anti-GFAP antibodies and properties of GFAP preparation were described. The immunobloting data on specificity of obtained monoclonal antibodies are presented. A new method of GFAP immunoenzyme assay based on GFAP preparation and anti-GFAP antibodies was elaborated. Standardization of the immunoenzyme system was shown in tests for specificity, accuracy, and reproducibility.