[The FUS protein: Physiological functions and a role in amyotrophic lateral sclerosis].

Certain forms of amyotrophic lateral sclerosis (ALS) are associated with an altered compartmentalization of FUS and its aggregation in the cytoplasm of motoneurons. FUS is a DNA/RNA-binding protein that is involved in DNA repair and the regulation of transcription, splicing, RNA transport, and local translation. Two theories have been proposed to explain the mechanism of the pathophysiological process in ALS. The theories attribute degeneration of motor neurons to either loss or gain of FUS function. The review describes the main physiological functions of FUS and considers evidence for each of the theories of ALS pathogenesis.

[Effects of ischemia on the expression of neurotrophins and their receptors in rat brain structures outside the lesion site, including on the opposite hemisphere].

Neurotrophins stimulate the regeneration of neural tissue after lesions. It is also known that the sources of neurogenesis and cerebral function recovery are predominantly located in subcortical brain structures. The effects of ischemia on the expression of genes that encode neurotrophins (Bdnf, Ngf, Nt-3) and their receptors (TrkB, TrkA, TrkC, p75) in brain structures outside the lesion site were studied 3, 24, and 72 h after irreversible unilateral occlusion of the middle cerebral artery in rats. Changes in the mRNA expression of these genes were assessed by relative quantification using real-time RT-PCR. Sham surgery was found to stimulate the expression of genes that encode neurotrophins (Bdnf, Ngf) and their receptor (p75). It has been shown that ischemia influenced the expression of neurotrophins (Bdnf, Ngf, Nt-3) and their receptors (TrkB, TrkA, TrkC, p75) in brain structures outside the lesion focus, including the contralateral hemisphere. The downregulation of Bdnf and TrkB transcripts and Ngf and TrkA upregulation in the contralateral cortex on the first day of ischemia obviously reflected stress response. On day 3, Nt-3 transcription increased in all investigated structures outside the lesion focus. In the contralateral hemisphere, relative levels of TrkA and TrkC mRNA expression increased, while p75 expression decreased. Presumably, the observed changes in gene transcription serve to facilitate neuroplasticity and neural tissue regeneration.

[Characteristics of immune response and inflammatory reaction in atherothrombotic stroke and myocardial infarction]. / Osobennosti immunnogo otveta i vospalitel'noi reaktsii pri aterotromboticheskom insul'te i infarkte miokarda.

AIM: To study characteristics of ischemic tissue damage basing on the assessment of the correlations between markers of immune response, inflammation and apoptosis in patients with myocardial infarction (MI) and atherothrombotic stroke (AS). MATERIAL AND METHODS: Concentrations of matrix metalloproteinase-9 (MMP-9), immune complexes with cryogenic properties, soluble Fas-ligand, tumor necrosis factor alpha, interleukin-10 measured with ELISA as well as the activity of spontaneous apoptosis of mononuclear cells and surface expression of Toll-like receptors-4 and intracellular heat shock proteins measured with flow cytofluorometry were determined in the blood of 93 patients with the first AS and 94 patients with MI without concomitant inflammation in the 1st and 7th day of the disease. RESULTS AND CONCLUSION: Increased levels of the markers of immune response, inflammation, apoptosis and destruction of the extracellular matrix were identified at the beginning of MI and AS. The results provide the evidence that similar mechanisms may be involved in ischemic tissue damage. Multivariate analysis conducterd by of principal component analysis correlation matrix revealed the specificity of the relationships between all of these markers. This is the completely new approach to assessin the role and importance of defined parameters in the acute period of the myocardial ischemia and brain.

[A mice model of amyotrophic lateral sclerosis expressing mutant human FUS protein].

UNLABELLED: BACKGROUND AND ОBJECTIVE: Loss of conformation and function of sufficient number of proteins with high aggregation capacity plays an important role in the pathogenesis of many neurodegenerative disorders (NDD). Due to a recent discovery of new array of proteins with the capacity to form aggregates of nonamyloid type, new NDD models as well as a new level of understanding in vivo models which are already exist is needed. DNA/RN A binding proteins - FUS and TDP-43 play a crucial role in the pathogenesis of some forms of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia. The objective of the study was to develop a new ALS transgenic model. MATERIAL AND METHODS: In cell culture experiments, we studied mutant FUS proteins capable to form intracellular deposits morphologically similar to those observed in the autopsy material of ALS patients. RESULTS AND CONCLUSION: We created a transgenic mice line, in which a pathogenic form of human FUS protein was expressed in the nervous system. That led to the aggregation of FUS protein in spinal cord and motor neurons with the following degeneration and development of a phenotype, similar to the human ALS disease phenotype, in young grown-up animals. This neurodegenerative phenotype corresponds to a great number of clinical manifestations of human ALS and is an adequate transgenic model of the disease.

[The effect of semax and its C-end peptide PGP on Vegfa gene expression in the rat brain during incomplete global ischemia].

Vascular endothelial growth factor (VEGFA) is a hypoxia-inducible signal glycoprotein. VEGFA causes vascular endothelial cell growth and proliferation, that leads to the regeneration of vascular network in brain regions damaged by ischemia. However, this protein is involved in processes of inflammation and edema in early stages of ischemia. Synthetic peptide semax shows neuroprotective and anti-inflammatory properties and is actively used in the treatment of ischemia.We have previously shown that semax reduces vascular injury and activates the mRNA synthesis of neurotrophins and their receptors under global cerebral ischemia in rats. Here we have analyzed the effects of semax and its C-terminal Pro-Gly-Pro tripeptide upon Vegfa mRNA expression in different rat brain regions after common carotid artery occlusion. The animals were decapitated 30 min, 1, 2, 4, 8, 12, 24 h after the operation. It was shown that ischemia increases levels of Vegfa mRNA in the rat brain of animals (4 h after the occlusion--in the cerebellum, cerebral cortex and hippocampus, 8 h--in the cortex and hippocampus, and 24 h in the cortex). Semax treatment reduces Vegfa mRNA levels in the frontal cortex (4, 8 and 12 h after the occlusion) and hippocampus of ischemic rats (2 and 4 h). Effect of PGP on the Vegfa gene expression was almost negligible. Our results showed that semax prevents activating effect ofhypoxia on the Vegfa gene expression in early stages of global ischemia. Furthermore, increase in the level of mRNA Vegfa in the hippocampus (24 h after occlusion) perhaps reflects neuroprotective properties of this drug.

[Estrogens and brain].

Recent data upon molecular mechanisms of pleiotropic action of estrogens in human brain is presented in the article. Given detailed descriptions of properties of classical and membrane bound estradiol receptors, that maintain gene expression regulation, modulation of neurotransmittent systems and signal cascade activation in neuronal cells. Data upon regional distribution of estradiol receptor subtypes in the brain, their participation in main cell population function control (including progenitor cells) is given. Special attention is paid to estrogen participation in neurogenesis, inflammation and apoptosis regulation in central nervous system; in the control of formation and functioning of cerebral vessels.

Intravenous xenotransplantation of human placental mesenchymal stem cells to rats: comparative analysis of homing in rat brain in two models of experimental ischemic stroke.

Mesenchymal stem cells from human placenta obtained after term natural delivery were cultured and labeled with vital dye Dil of magnetic fluorescing microparticles. The labeled cells were transplanted intravenously to rats with occlusion of the median cerebral artery. Penetration of cells through the brain-blood barrier and their distribution in the brain of experimental animals were studied on serial cryostat sections. Two models of cerebral artery occlusion associated with different traumatic consequences were used. The efficiency of crossing the blood-brain barrier by transplanted cells, the number of mesenchymal cells attaining the ischemic focus and neurogenic zones, and the time of death of transplanted cells largely depended on the degree and nature of injury to the central nervous system, which should be taken into account when planning the experiments for evaluation of the effects of cell therapy on the models of neurological diseases and in clinical studies in the field of regenerative neurology.

[Dynamics of higher mental functions and complications in the early post-operative phase of carotid endarterectomy].

Authors conducted a clinical-instrumental examination of 97 patients in the post-operative phase of reconstructive surgery of carotid arteries. Evaluation of neurological status, Luria's syndrome neuropsychological analysis for assessment of higher mental functions dynamics (HMF) were carried out. Pre-operative ultrasound duplex scanning of brachio-cephalic arteries and investigation of cerebral perfusion using spiral computed tomography (SCT-perfusion) were done before and 1 day after the surgery. The frequency of intrasurgery stroke in the area of the operated carotid artery was 2.1%. Pre-operative SCT-perfusion revealed the syndrome of cerebral hyperperfusion in 3.1% of patients with higher (>1 s) difference between the average time of blood transition in temporal areas. Dynamics of HMF was the key clinical indicator that allowed to evaluate the course of nearest post-operative phase and to compare it with an area of operated vessel. Changes of HMF were characterized by the decrease in dysfunction of the hemisphere ipsilateral to the side of operation or had differently directed character. The aggravation of HMF disturbances was found in 40% of patients and was represented by syndromes with different topical location. Causes and mechanisms of the development of these changes are discussed.

[The analysis of risk factors for acute stroke in Dagestan].

Results of the first epidemiologic survey of cerebral stroke in Dagestan carried out in 2009 using a method of population register are presented. Arterial hypertension was on the first place (98.5%) among risk factors. Other risk factors were determined as follows: cardiac diseases (24.2%), cardiac fibrillation (14.5%), lipid profile dysfunction (10.6%), smoking (8.7%), diabetes mellitus (8.6%), stress (7.5%), history of myocardial infarction (3.6%).

[The effect of semax and its C-end peptide PGP on expression of the neurotrophins and their receptors in the rat brain during incomplete global ischemia].

Neurotrophins regulate key function of nervous tissue cells. Analysis of neurotrophins mRNA expression is an appropriate tool to assess therapeutic efficiency of the anti-stroke drugs. We have analyzed the effect of synthetic peptide semax and its C-terminal Pro-Gly-Pro tripeptide upon mRNAs expression of neurotrophins Ngf, Bdrf, Nt-3 and their receptors TrkA, TrkB, TrkC, p75 in rat frontal lobes, hippocampus and cerebellum after bilateral common carotid artery occlusion. The animals were decapitated 30 min, 1, 2, 4, 8, 12, 24 h after the operation. The mRNA expression of neurotrophins and their receptors was assessed by relative quantification using real-time RT-PCR. Our showed that ischemia causes a significant decrease in gene expression in the hippocampus. Semax and PGP affected the expression of neurotrophins and their receptors predominantly in the frontal cortex and hippocampus of the ischemized animals. In the frontal cortex, Semax treatment resulted in a decrease of mRNA level of receptors, while PGP treatment increased the level of these mRNA. Maximal neuroprotective effect of both peptides has been observed in the hippocampus 12 h after occlusion. A decrease of gene expression of neurotrophins and their receptors caused by the occlusion was overcome by Semax and PGP. These results clarify the semax mechanism of and present certain features of mRNA's expression of neurotrophins and their receptors in experimental conditions.

[Models of neurosurgical service for patients with hemorrhagic stroke].

Authors have conducted an analysis of surgical treatment of patients with hemorrhagic stroke (HS) in Moscow neurosurgical in-patient clinics. In 2 most advanced neurosurgical departments out of 11 studied, only 61% of necessary neurosurgical care is carried out. There are three models of specialized neurosurgical care for patients with HS in large cities. The most effective is a model which comprises a mobile consulting neurosurgical team for selection of patients for surgery and their transport. The causes of insufficient amount of surgical interventions in neurosurgical in-patient clinics are specified. The authors suggest the algorithm of HS patients' selection for surgery.

[An experimental model of focal brain ischemia in rats based on the endovascular microsurgery].

An endovascular model of focal brain ischemia in rats with controlled duration of reperfusion of 1h has been studied. The reliability of experimental model for localization and volume of ischemic lesion (basal ganglia, hippocampus, parietal-temporal regions; the volume of lesion focus on MRI T2-weighted image (T2-WI) in the first day after brain ischemia 146.4 +/- 44.7 mm3) has been worked out. We conducted the clinical monitoring including assessment of neurological deficit, behavioral tests and performed MRI of the brain on 1st, 5th and 10-17th days after the occlusion of middle cerebral artery (MCA), including regimes of diffusion-weighted image (DWI) and T2-WI, and carried out a histological study of the brain. The regress of neurological deficit was seen on the 14th day after surgery. Behavioral tests revealed the reduction of movement activity of animals in the case group 7 days after the MCA occlusion. Compared to the first day after surgery, the decrease of volume lesion focus on DWI and T2-WI was observed on the 5th day (p < 0.05) as well, with the following decrease to the 10-14th days. The histological picture in the lesion foci corresponded to the brain infarction on the 5th and 14th days after surgery.

Mechanisms of positive effects of transplantation of human placental mesenchymal stem cells on recovery of rats after experimental ischemic stroke.

Mesenchymal stem cells isolated from human placenta and in vitro labeled with fluorescent magnetic microparticles were intravenously injected to rats 2 days after induction of focal cerebral ischemia (endovascular model). According to MRT findings, transplantation of mesenchymal stem cells led to an appreciable reduction of the volume of ischemic focus in the brain. Two or three weeks after transplantation, labeled cells accumulated near and inside the ischemic focus, in the hippocampus, and in the subventricular zone of both hemispheres. Only few human mesenchymal stem cells populating the zone adjacent to the ischemic focus started expressing astroglial and neuronal markers. On the other hand, transplantation of mesenchymal stem cells stimulated proliferation of stem and progenitor cells in the subventricular zone and migration of these cells into the ischemic zone. Positive effects of transplantation of these cells to rats with experimental ischemic stroke are presumably explained by stimulation of proliferation of resident stem and progenitor cells of animal brain and their migration into the ischemic tissue and adjacent areas. Replacement of damaged rat neurons and glial cells by transplanted human cells, if it does take place, is quite negligible.

Use of bone marrow mesenchymal (stromal) stem cells in experimental ischemic stroke in rats.

The effects of human mesenchymal stem cells on neurological functions and behavioral reactions of animals and on damaged brain tissue were studied on the model of focal cerebral ischemia in rats. Homing and differentiation of transplanted mesenchymal stem cells were also studied. Significant regression of neurological disorders after cell transplantation was noted, no appreciable shifts were detected by magnetic resonance tomography. Homing of transplanted cells was detected mainly in the zone of focal ischemia. Some cells died, others exhibited signs of differentiation into neurons and glia.

[An effect of perindopril on the level of tumor necrosis factor-alpha and matrix metalloproteinase-9 in peripheral blood in the acute period of atherothrombotic ischemic stroke and myocardial infarction].

Twenty-nine patients with acute atherothrombotic ischemic stroke and 36 patients with acute Q-wave myocardial infarction have been studied. Each group has been stratified into 2 subgroups: patients of subgroups A received an ACE inhibitor perindopril in the complex therapy from the 1st day of disease. Patients of subgroups B were not assigned to this drug. Along with routine tests, the level of tumor necrosis factor-alpha and matrix metalloproteinase-9 (MMP-9) measured with ELISA using test-systems (BCM Diagnostics, USA) and reagents (R&D, England) have been determined. The administration of perindopril did not cause side-effects, including arterial hypotonia after the first dosage, in patients in the acute period of atherothrombotic ischemic stroke and myocardial infarction. Perindopril may decrease the activity of MMP-9 in these patients and produces an anticytokine effect. Some similar mechanisms of ischemic lesions of the heart and the brain and a commonness of biochemical "response" to the same medical intervention (the administration of an ACE inhibitor perindopril) in patients of both groups were found. The results support the pathogenetic validity of perindopril therapy in the secondary prevention of ischemic stroke and myocardial infarction.

[New possibilities of neuroprotection in the treatment of ischemic stroke].

A multicenter prospective double-blind placebo-controlled study was carried out in 62 patients with hemisphere ischemic stroke of atherothrombotic or cardiothrombotic character. Thirty-two patients, a main group, received intramuscular injections of cortexin, a domestic drug, in dosage 20 mg daily during 10 days from the first 6 hours from the development of symptoms of stroke, and 30 patients, a control group, received basic treatment and intramuscular injections of placebo in dosage 20 mg daily during 10 days. Patient's state was assessed before treatment and on 3rd, 7th, 11th and 28th days using the NIHSS scale for assessment of disease severity, intensity of focal neurologic deficit and dynamics of clinical parameters and the modified Rankin scale and Barthel index as a functional outcome measures. The data obtained suggest an efficacy of cortexin in the treatment of patients with acute ischemic stroke as compared to placebo. Safety of the drug was confirmed in the study.

[Oxidative stress and oxygen status in ischemic stroke].

A dynamic assessment of oxygen status of the arterial blood, activity of antioxidant system enzymes (AOS), succinatedehydrogenase (SDG), mitochondrial alpha-glycero-phosphate-dehydrogenase (alpha-GPDH) and alkaline phosphatase (AP) as well as concentrations of reduced glutathione (GSH) and secondary products of lipid peroxidation reacting with thiobarbituric acid (PLPRTA) has been carried out in patients at the acute stage of ischemic stroke of hemispheric location. Relative hyperoxia as a result of the hyperventilation syndrome was mostly pronounced on day 1 and 3. At the same time, a reduced activity of AOS system and an increase of PLPRTA concentration have been observed from the 1st day after stroke. There were also a decrease of the SDG activity and a marked (2,8 fold) increase of the alpha-GPDH activity as compared to the controls. A decrease of the AP leukocyte activity in the peripheral blood to day 7 after stroke makes possible a prognosis of good functional rehabilitation to the 21st day of the disease. Therefore, the results of the study suggest that the development of oxidative stress in patients with ischemic stroke is caused by tprimary disruption of bioenergetic processes during the reduction of AOS activity.

[The study of chronic partial denervation and quality of life in patients with motor neuron disease treated with semax].

The study of chronic partial denervation (CPD) and quality of life was carried out in 27 patients with definite, probable and possible diagnosis of motor neuron disease (MND) treated with semax (1% solution). The needle electromyography (EMG) was performed thrice with short-term 2 month interval (60 days before enrollment and on day 1 and day 48 of the study) in three muscles on bulbar, cervical and lumbosacral levels on the less affected side. According to Revised El-Escorial Criteria (1998) the needle EMG for diagnostic purposes was also performed in two muscles on the cervical and lumbosacral levels on the more affected side along with stimulation electroneuronmyography of motor and sensory fibers of the peripheral nerves of neck, upper and lower extremities. The open-label clinical trial of Semax (1% solution) was conducted in sequential groups of patients. The drug was administered intranasally in two 10-day-long courses with 2-weeks break in daily dose of 12 mg. Sixty days before enrollment, and on days 1, 10, 24, 34 and 48, patients were assessed by the Norris ALS, the ALS Functioning Rating Scale and the ALSAQ-40 quality of life in the ALS scale. It was shown that CPD on the early as well as on the late stages was characterized by forward-backward, but not unidirectional course, that did not allow to recommend the follow-up needle EMG with short-term interval for evaluation of drug efficacy monitoring. Early CPD stages were characterized by forward-backwards fluctuations reflecting the compensatory reinnervation process (a phenomenon of exchange of muscle fibers, more rational in view of reinnervation, between adjacent motor units) whereas on the late CPD stages these forward-backwards CPD fluctuations reflected the processes of progressive deterioration of muscle fibers and secondary demyelination of large motor axons. Semax (1% solution) does not influence either the course of CPD or the dynamics of clinical estimates, in particular the terms of ensuing marked functional deficits on bulbar, cervical and lumbosacral levels of segmental innervation. However, 1% semax significantly improves the total estimate of life quality due to the improvement of emotional state and motivation in MND patients with the maximal effect on day 10. This finding suggests that it is feasible to administer 1% semax in complex MND palliative therapy.