Methods of Generation of Induced Pluripotent Stem Cells and Their Application for the Therapy of Central Nervous System Diseases.

The use of induced pluripotent stem cells (IPSC) is a promising approach to the therapy of CNS diseases. The undeniable advantage of IPSC technology is the possibility of obtaining practically all types of somatic cells for autologous transplantation bypassing bioethical problems. The review presents integrative and non-integrative methods for obtaining IPSC and the ways of their in vitro and in vivo application for the study and treatment of neurological diseases.

Distribution and Migration of Human Placental Mesenchymal Stromal Cells in the Brain of Healthy Rats after Stereotaxic or Intra-Arterial Transplantation.

Human placenta mesenchymal stromal cells were injected to healthy rats either stereotaxically into the striatum or intra-arterially through the internal carotid artery. Some cells injected into the brain migrated along the corpus callosum both medially and laterally or concentrated around small blood vessels. A small fraction of MSC injected intra-arterially adhered to the endothelium and stayed inside blood vessels for up to 48 hours mostly in the basin of the middle cerebral artery. Neither stereotaxic, nor intra-arterial transplantation of mesenchymal stromal cells modulated the proliferation of neural stem cells in the subventricular zone of the brain, but stereotaxic transplantation suppressed activation of their proliferation in response to traumatization with the needle.

Comparative Analysis of the Effects of Intravenous Administration of Placental Mesenchymal Stromal Cells and Neural Progenitor Cells Derived from Induced Pluripotent Cells on the Course of Acute Ischemic Stroke in Rats.

We compared the effects of placental mesenchymal stromal cells and neural progenitor cells derived from induced human pluripotent cells after their intravenous administration to rats in 24 h after transitory occlusion of the middle cerebral artery. The therapeutic effects were evaluated by the dynamics of animal survival, body weight, neurological deficit, and the volume of infarction focus in 7, 14, 30, and 60 days after surgery. Intravenous injection of neural progenitor cells produced a therapeutic effect on the course of experimental ischemic stroke by increasing animal survival in the most acute period and accelerating compensation of neurological deficit and body weight recovery. Neural progenitor cells were more effective than mesenchymal stromal cells from human placenta. The effectiveness of intravenous transplantation of neural progenitor cells in the model of occlusion of the middle cerebral artery is shown by us for the first time, although the therapeutic effect of their direct transplantation into the brain has already been described.

[Dynamics of perfusion CT parameters in the early acute phase of ischemic stroke].

Perfusion computed tomography (CT) allows to evaluate brain perfusion and provides additional information about local cerebral hemodynamics in stroke patients. Twenty-seven patients underwent CT-perfusion in the early acute phase of ischemic stroke. Significant differences between ischemic lesion and contralateral hemisphere on CBF and MTT scans during the first 3 hours after stroke were revealed. In the following 24 h, the differences were seen in all perfusion parameters. CT-perfusion allows to reveal perfusion parameters associated with stroke severity and outcome that is relevant for prescription of reperfusion treatment beyond the boundaries of 3-4.5h "therapeutic window".

[Mathematical modeling of group of neurons and astrocytes in ischemic stroke].

The mathematical model of group of neurons and astrocytes in the ischemic stroke is discussed. The model includes the description of synaptic signal transmission between the neurons, calcium signaling in astrocytes, ionic currents between the cells and extracellular space and the diffusion through extracellular space. The new modeling approach, based on the creation of appropriate graph, is developed to describe the structure of the system. Using the model, we have analyze the influence of inhibitory synapses on system functioning. We also have shown how the use of medications supporting the activity of inhibitory synapses affects the system behavior in ischemic stroke.

[Computer tomographic and laboratory predictors of clinical outcomes of acute ischemic stroke patients after the systemic thrombolytic therapy].

Today the systemic thrombolytic therapy with recombinant tissue plasminogen activator (rt-PA) is the most effective treatment of ischemic stroke. We included 116 patients with stroke, 82 received rt-PA, 34 were enrolled to the control group. Median NIHSS score was 16,2 +/- 5,4 in the main group and 16,2 +/- 5,4 in the control group. The trial revealed the prognostic significance of some early CT sings of ischemic brain damage for the hemorrhagic transformation and functional outcomes to the 90th day. It was shown that hyperfibrinogenemia (over 450 mg/dl) might be a risk factor of the hemorrhagic transformation. The excess of segmental leukocytes over 78% might be a predictor of the lethal outcome. The initial activity of tissue plasminogen inhibitor (first type) over 11,65 IU/L might be a risk factor of the vascular reocclusion after successful thrombolysis. The analysis of characteristic curves of leukocyte elastase revealed the informativeness of its initial values in the prognosis of reocclusion and hemorrhagic transformation.

[Gestagens and the brain].

Modern data on molecular mechanisms underlying intracerebral pleutropic action ofgestagens (progesterone, its metabolites, and synthetic gestagens) are reviewed. Properties of classical and membrane-bound progesterone receptors involved in regulation of gene expression, modulation of neurotransmitter systems, and activation of signal cascades in neurons are described along with data of regional distribution of progesterone receptor subtypes in brain, metabolism, functional activity, and mechanisms of action of progesterone metabolites. Special attention is given to analysis of progesterone and metabolite neuroprotective activity in CNS, such as anti-inflammatory and antiapoptotic effects, and participation in neurogenesis regulation. Clinical application of progesterone-based drugs is discussed with reference to the treatment of brain injuries and prospects for extension of indications for their use in neurologic practice.

[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.

[Experimental hemorrhagic stroke: the study of neuropeptides (MIF, selank) in the intraperitoneal injection].

The aim of this study was to examine Taftsin derivates--macrophage inhibitory factor (MIF, Thr-Lys-Pro) and heptapeptide selank (Thr-Lys-Pro-Arg-Pro-Gly-Pro) in the model of experimental intracerebral hemorrhage in rats. The double autologous blood injection in the basal nucleus was used as a model of intracerebral hemorrhage. Animals ware randomly divided into three groups--the control group (n = 5) was treated with saline, the second group (n = 5) was injected with MIF in dose 150 mkg/ kg/day, the third group (n = 5) received Selank in dose 300 mkg/kg/day. Intraperitoneal injection of peptides was used. Body weight assessment, neurological examination and brain MRI were performed in 24, 72 hours and 10 days after the hematoma formation. The effect of neuropeptides on the functional restoration in animals, in the absence of the effect on hematoma volume and perifocal edema, was found. The significant reduction of perifocal edema and hematoma volume was observed in the 10th day after the hematoma formation in all experimental groups (p < 0.05). Only the control group of animals showed the significant (p < 0.05) weight loss in the 3rd day after the operation. The rate of neurological deficit was different: the significant improvement assessed with Menzes and limb placing test scales was seen only in the groups treated with neuropeptides in the 10th day.

[MRI semiotics features of experimental acute intracerebral hematomas].

The aim of this study was to assess the possibility of revealing intracerebral hematomas (ICH), using MRI, within the first hours after onset and to determine their MRI semiotics features. Thirty animals with experimental ICH were studied. A method of two-stage introduction of autologous blood was used to develop ICH as human spontaneous intracranial hematomas. Within 3-5h after blood introduction to the rat brain. The control MRI was performed in the 3rd and 7th days after blood injections. ICH were definitely identified in the first MRI scans. The MRI semiotics features of acute ICH and their transformations were assessed. The high sensitivity of MRI to ICH as well as the uniform manifestations in all animals were shown. In conclusion, the method has high specificity for acute ICH detection.

[High efficacy of selective intra arterial thrombolysis in ischemic stroke patients with large-vessel occlusion].

Thrombolysis is the most effective treatment of acute ischemic stroke that increases the proportion of patients with good clinical outcome. Selective intra-arterial thrombolysis (IAT) can be used in a wider therapeutic window (up to 6-8 hours) under the angiographic control with tailoring of fibrinolytic dose. The results of IAT by a recombinant plasminogen activator in 2 patients are presented. Patient 1, male, 55 years old, with M1 cerebral middle artery (CMA) occlusion and with NIHSS score 13 on admission was treated by IAT after 7 hours of stroke onset. Recanalization was observed 40 minutes after IAT started. The NIHSS score was 8 after IAT. Patient 2, female, 64 y.o., with NIHSS score 20 on admission and C7 internal carotid artery and M1 CMA occlusion was treated by IAT. Recanalization was observed 60 minute after the beginning of IAT, with NIHSS score being decreased to 14. These clinical cases demonstrate the high efficacy of intra arterial thrombolysis in ischemic stroke patients.

[Systemic thrombolysis in ischemic stroke].

Intravenous recombinant tissue plasminogen activator (rt-PA) is an approved and effective treatment for acute ischemic stroke within 3 hours of symptoms onset. The results of the first-ever implementation of the thrombolysis in the Russian Federation are presented. Fourteen stroke patients received rt-PA in dose of 0.9 mg/kg (10% as a bolus during 1 minute followed by infusion), the mean "door-to-needle" time was 50 +/- 23.6 min. The thrombolysis resulted in the clinical improvement by the end of the first day after stroke onset in 9 (64.2%) patients, with a dramatic regress of neurological deficit (the decrease of the NIHSS scores = 4) in 3 (21.4%) of them. Hemorrhagic transformation was observed in 6 (42.8%) patients, however in 5 cases it was asymptomatic and was found only in the repeated CT study to the end of the first day. Three months after stroke onset, 7 (50%) patients scored 0-1 on the modified Rankin scale, 1 patient had 4 scores and 1 patient died. Thus the thrombolysis can change the stroke management and significantly improves the outcome.

[A randomized, double-blind, placebo-controlled study of Cerebrolysin safety and efficacy in the treatment of acute ischemic stroke]. / Randomizirovannoe dvoinoe slepoe platsebo-kontrolirumoe issledovanie bezopasnosti i effektivnosti tserebrolizina dlia lecheniia ostrogo ishemicheskogo insul'ta.

The aim of the study was to assess safety and efficacy of the neuroprotective drug Cerebrolysin in acute ischemic stroke. Thirty-six patients with ischemic stroke in carotid artery territory aged 45-85 years, were eligible for inclusion in the trial if they were admitted to the hospital within the first 12h after stroke onset. Patients were randomly and blindly assigned to placebo (n = 12) or 1 or 2 dosages of Cerebrolysin: 10 ml/d (n = 12) and 50 ml/d (n = 12) for 10 days with concomitant standard basic treatment in each group. A quantitative time-related analysis of the dynamics of neurological deficit revealed the tendency towards acceleration of improvement assessed by the Clinical Global Impression Scale and NIHSS in both Cerebrolysin groups by 30 day of the treatment. The significant reduction in the volume of MRI ischemic focus was shown in both Cerebrolysin groups (p < 0.05 vs Placebo) on day 3. Acute pharmacological test revealed a decrease (p < 0.05 vs Placebo) of the size and spread of delta and theta foci in 72.7% patients, receiving 50 ml/d of Cerebrolysin. In none of the cases, Cerebrolysin treatment provoked any paroxysmal activity on EEG. The trial demonstrated safety, efficacy and good tolerability of hige-dose Cerebrolysin in the treatment of ischemic stroke.

Connection between p53 gene Bam HI RFLP polymorphism with the volume of brain infarction in patients with carotid atherothrombotic ischemic stroke.

PURPOSE: The aim of our investigation was to study the connection between p53 gene Bam HI RFLP polymorphism and the brain infarction volume in patients with atherothrombotic ischemic stroke that could highlight certain genetic aspects of the individual sensibility of brain tissue to acute ischemia. MATERIALS AND METHODS: Diallelic Bam HI RFLP polymorphism in 5' flanking region p53 gene was studied in 96 patients with carotid atherothrombotic stroke from Moscow population. Magnetic resonance imaging was conducted on day 7 after the stroke onset. The manual morphometry and "Osiris" morphometric hardware (by the Hospital of the University of Geneva) were used for assessment of the infarction volume. RESULTS: The predominance of small-size infarctions (< 40 cm3) was revealed in patients with (-/-) Bam HI RFLP p53 genotype versus patients with (-/+) (X2 = 19.7; P < 0.001) and (+/+) (X2 = 12.288; P < 0.001) genotypes. According to the Bayesian's statistics, in patients with (-/-) p53 Bam HI genotype the development of a small-size infarction in atherothrombotic ischemic stroke can be prognosticated with probability more than 65%. CONCLUSIONS: A significant association between p53 gene Bam HI RFLP polymorphism and the infarction volume was found in patients with carotid atherothrombotic stroke from Moscow population. These results additionally confirm that apoptosis plays an important role in the formation of ischemic brain lesion and that drugs with anti-apoptotic properties may prove beneficial in stroke patients.

[The dynamics of recovery processes in ischemic stroke as shown by comparison of clinical and tomographic indices]. / Dinamika vosstanovitel'nykh protsessov pri ishemicheskom insul'te po dannym sopostavleniia klinicheskikh i tomograficheskikh pokazatelei.

The aim of this study was to assess the relation between MRI-changes of the brain infarction (BI) and neurological status in patients with hemispheric ischemic stroke. Clinical and MRI examinations were performed in 70 patients (aged from 46 to 89 years, 37 females, 33 males) 1, 3, 7-10, 14 and 21-30 days (in all cases) and 2, 6 and 12 months (n=29) after stroke onset. On FLAIR-images the criteria of assessment of cystic and gliosis transformation were determined. The tissue contrast of BI is assigned mainly to brain edema and hemorrhagic transformation, the cystic and gliosis transformation visualized in 1-2 months after stroke onset. The ratio between cystic and gliosis transformation correlated with clinical dynamics and the size of BI.

[Association between Bam HI RFLP p53 gene polymorphism and brain infarction volume in patients with atherothrombotic ischemic stroke]. / AssotsiatsiiaBam HI RFLP polimorfizma gena p53 s ob"emom infarkta mozga u patsientov s aterotromboticheskim ishemicheskim insul'tom.

A significant association between Bam HI RFLP p53 gene polymorphism and brain infarction volume in patients with atherothrombotic ischemic stroke (AIS) from Moscow population was found. Biallelic Bam HI polymorphism in exon 5 of p53 gene was studied in 96 AIS patients, 100 their healthy siblings, 41 patients with chronic ischemic disease and 42 their healthy siblings. Magnetic resonance tomography with morphometric program "Osiris" (the Hospital of the University, Geneva) for infarction volume measuring was used on day 1 and 7 after stroke onset. The patients with (-/-) p53 Bam HI genotype had the smaller brain infarction sizes (< 40 cm3), comparing to those with (-/+) (chi 2 = 19.7; p < 0.001) and (+/+) (chi 2 = 12.3; p < 0.001) genotypes. According to Bayes statistics, development of smaller infarction size in AIS may be predicted with over than 65% probability in the patients with (-/-) p53 Bam HI genotype. A significant association between p53 Bam HI polymorphism and infarction volume in patients with carotid atherothrombotic stroke confirms an important role of apoptosis in ischemic brain lesions formation that demands temporary antiapoptotic influence on patients with stroke.

[Disrupted circle of Willis and crisis course of arterial hypertension]. / Razomknutyi villiziev krug i "krizovoe" techenie arterial'noi gipertenzii.

Two patients, aged 17 and 55 years, with partly or completely open circle of Willis were examined using cerebral magnetic resonance angiography. A role of anatomical structure of circle of Willis and other anastomotic peculiarities of brain blood supply in cerebral vascular crises development is demonstrated. The latter can proceed as hypertensive ones on the background of emotional overloading and also reflect other features of brain blood supply in its clinical picture e.g. an absence of one of spinal arteries on the background of atherosclerosis and arterial hypertension.