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Various rodent models have been developed for examining either focal or global cerebral ischemia so far, but the most used model is the rat middle cerebral artery occlusion by intraluminal filaments, followed by reperfusion, and 2,3,5-triphenyltetrazoliumchloride or Cresyl Violet/Nissl staining for evaluation of the region of interest. Like other surgical procedures, this one is prone to various complications (excessive bleeding, vascular or nerve lesions) and failure related to the surgical technique. We focused on detailed surgical techniques, along with data from literature, in order to reduce the complications and increase the chance of experiment success. Magnetic Resonance Imaging evaluation with 3D reconstruction of the ischemic area opens new perspectives on investigations into the ischemic brain, having the advantage of being noninvasive as an alternative to gold histological standard for measuring the infarct size, being the only one accesible in surviving subjects.
The acute phase response follows tissue injury and contributes to its exacerbation with pro-inflammatory and pro-thrombotic mechanisms. Acute phase proteins promote erythrocyte aggregation and falling, with the result that the erythrocyte sedimentation rate (ESR) is a measure of the acute phase response. As the acute phase response accompanies ischaemic brain damage, we studied ESR values in patients within the first 24 hours of ischaemic stroke and evaluated whether these values may be related to the volume of anatomically relevant single hemispheric brain computed tomography (CT) areas observed at the same period, indicating early stroke-related cerebral changes. We observed an increase in ESR in stroke patients and a positive correlation between the ESR values and the volume of early brain CT hypodense areas. The results suggest that elevation in ESR values is observed soon after a stroke and may reflect the relationship between the degree of acute phase response in the early phase of ischaemic stroke and the extent of local brain damage.
Tumor necrosis factor alpha (TNF-alpha) is a protein of a cellular origin belonging to a group of proinflammatory cytokines. A rapid overproduction of TNF-alpha in a cerebral post-ischemic inflammatory response leads to the stimulation of adhesive molecules expression with subsequent accumulation of leukocytes in the ischemic focus, which is preceded by their adhesion and migration. The TNF-alpha proinflammatory activity results mainly in extending the area of the brain infarct, which brings about negative clinical implications. Being the final morphological effect of ischemic stroke, TNF-alpha appears also to contribute to neuronal necrosis by its involvement in the process of apoptosis as well as in the death of neurons. The present study describes and discusses mainly the contribution of TNF-alpha to the formation of ischemic focus in the brain.
Stroke-induced inflammatory reaction leads to the accumulation of leukocytes in the brain ischaemic region, where they exert a detrimental effect - promotion and extension of cerebral damage. Intracerebral infiltration of peripheral blood leukocytes requires prior endothelial-leukocyte interactions that are mediated by such cell surface proteins as adhesion molecules. Among adhesion molecules, it is the immunoglobulin gene superfamily (IgSF) that is responsible for strong attachment and transendothelial migration of leukocytes. The principal members of IgSF are: intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and platelet endothelial cell adhesion molecule-1 (PECAM-1). In this review the following issues were described and discussed: an increased expression of ICAM-1 and VCAM-1 in ischaemic brain as well as a detection of their soluble(s) forms in sera of stroke victims. The presented data suggest the involvement of both ICAM-1 and VCAM-1 in the sequence and timing of the infiltration of leukocytes into the brain ischaemic zone after stroke. They have also revealed changes in serum concentrations of sICAM-1 and sVCAM-1 that are characteristic for stroke. Recently, increase in sPECAM-1 levels in serum and cerebrospinal fluid (CSF) has been shown within 24 h of the onset of stroke, having indirectly suggested involvement of the molecule in the inflammatory events during the early phase of stroke.
Stroke is a major cause of adult disability that poses an enormous healthcare burden. Effective pharmacotherapy for stroke remains an unmet need. Development of restorative therapies has been identified as a potential alternative in stroke. Emerging understanding of brain repair and plasticity mechanisms have revealed therapeutic targets including inhibition of axonal sprouting (e.g., Nogo, MAG), altered perilesional GABA and glutamate receptor signaling, endogenous neurogenesis and angiogenesis. The main advantage with restorative therapies is the delayed treatment after acute necrotic cell death, when patients are stable. In addition, restorative therapies can be combined with intensive rehabilitation and medication for poststroke complications to further facilitate recovery process. The problem with patient studies is, however, that many pharmaceutical companies have scaled down their stroke programs, because of failures with neuroprotective compounds. We should convince industry that restorative drugs target completely different mechanisms with extended therapeutic time window offering an attractive approach to help stroke patients.
Background. Ventilation volume is a parameter used mainly for determining oxygen consumption of fish. The aim of the present work was to determine the ventilation volume of carp, under conditions of pure, aerated water. Materials and Methods. Stroke volume and breathing rate of carp representing three size groups (258.7 ± 40.1 g, 449.3 ± 39.6 g, and 663.2 ± 32.3 g) were studied within the temperature range of 10–25°C. Results. At 10°C the stroke volume of carp weighing 200–300 g was on average 2.25 ± 0.63 ml per 1 breath. This parameter in fish weighing 400–500 g was 2.70 ± 0.12 ml·per 1 breath, while in fish attaining 600–700 g it amounted to 3.22 ± 0.41 ml·per 1 breath. The breathing rate of all size groups ranged from 46.2 to 47.4 ± 8.51 cycles per min. A statistically significant increase of the stroke volume was recorded in all size groups at 15°C. At 20°C the increased stroke volume was accompanied by accelerated breathing rate. The temperature increase from 20 to 25°C did not cause any further increase of either breathing rate or stroke volume. Conclusion. The temperature-related regulation of the ventilation volume in carp is a two-step process. At 10–15°C the increased water volume pumped through the gills was achieved by an increased breathing depth (stroke volume). A further increase of the ventilation volume at 15–25°C resulted from acceleration of the breathing rate.
Although there are some in vitro evidence that angiotensin II (Ang II) may promote thrombosis, there is still no data concerning effect of Ang II on arterial thrombus formation. In the present study we have investigated the influence of Ang II on electrically induced arterial thrombosis in a common carotid artery of renovascular hypertensive rats. Furthermore, we examined if Ang II effect is mediated via AT1 receptor. We measured some coagulation and fibrinolytic parameters at the same time. Since platelets play crucial role in the initiation of arterial thrombosis their contribution in the mode of Ang II action was also determined. Intravenous infusion of Ang II caused significant increase in arterial thrombus weight, which was reversed by losartan, selective AT1 receptor antagonist. The prothrombotic effect of Ang II was accompanied by increase in haemostatic and decrease in fibrinolytic potential of rat plasma. While number of data has clearly demonstrated that Ang II can augment human platelets aggregation, at least in rats, platelets were not involved in the mechanism of Ang II action. Our study shows that Ang II via AT1 receptor accelerates arterial thrombosis in renovascular hypertensive rat, therefore may be considered as a risk factor of myocardial infarction or stroke.
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The incidence of sleep apnea in patients with stroke or transient ischemic attack

58%
Disorders of breathing during sleep are defined as cessation or reduction of air flow thorough the upper airway, accompanied by a decrease of oxygen saturation. The results of many studies underline the association between sleep-disordered breathing (SDB) and cerebrovascular disorders. SDB, mostly obstructive sleep apnea syndrome (OSAS), is believed to be an independent risk factor of stroke and is related to poor outcome and increased long-term stroke mortality. The present study evaluated the frequency of SDB in patients with stroke or transient ischemic attack transient ischemic attack. We studied 43 patients (mean age 68.5 ±11.0), which included 35 males and 8 females, with acute stroke (n=37) and transient ischemic attack (n=6). The assessment included body mass index (BMI), age, cardiovascular risk factors, and localization of stroke. All patients underwent all-night screening for SDB with a portable 8-channel recorder. The apnea/hypopnea index (AHI) for the whole group was 13.3 ±15.2. AHI <5 was found in 16 patients. Overall, SDB was present in 27 (62.8%) patients with stroke and transient ischemic attack, stratified into those with AHI 5-10, (10 patients), 10-20 (8 patients), and AHI>20 (9 patients). In 15 patients, there was an increase in AHI 5 on assuming the supine position. The patients’ mean BMI was 27.8 ±4.7. The analysis of BMI, age, and localization of stroke was not sufficient to identify patients with high risk for SDB. We submit that overnight screening for SDB should be routinely performed in every patient after stroke and transient ischemic attack and it should become a diagnostic tool in neurological departments.
Human adipose-derived stem cells (huADSC) were generated from fat tissue of a 65-year-old male donor. Flow cytometry and reverse transcription polymerase chain reaction (RT-PCR) analyses indicated that the huADSC express neural cell proteins (MAP2, GFAP, nestin and β-III tubulin), neurotrophic growth factors (BDNF and GDNF), and the chemotactic factor CXCR4 and its corresponding ligand CXCL12. In addition, huADSC expressed the characteristic mesenchymal stem cell (MSC) markers CD29, CD44, CD73, CD90, CD105 and HLA class I. The huADSC were employed, via a right femoral vein injection, to treat rats inflicted with experimental intracerebral hemorrhage (ICH). Behavioral measurement on the experimental animals, seven days after the huADSC therapy, showed a significant functional improvement in the rats with stem cell therapy in comparison with rats of the control group without the stem cell therapy. The injected huADSC were detectable in the brains of the huADSC treated rats as determined by histochemistry analysis, suggesting a role of the infused huADSC in facilitating functional recovery of the experimental animals with ICH induced stroke.
Background. Many scientific reports have shown a decrease in total cerebrovascular disease (CeVD) mortality over the past few decades, but too little attention has been paid to premature mortality. CeVD accounted for 22.5% and 17.8% of premature cardiovascular disease deaths in Poland, in 2000 and 2016, respectively. Objective. The aim of the study was to analyse premature CeVD mortality in the Polish population in the recent years, the dynamics of its changes and the potential factors that may have contributed to the decline in mortality. The main goal of the study was to overview the levels and trends in premature CeVD mortality with an emphasis on haemorrhagic, ischaemic and unspecified (not specified as haemorrhagic or ischaemic) stroke. Material and methods. The analysis was based on a database of the Central Statistical Office of Poland and included data from 2000-2016 on premature cerebrovascular deaths occurring between 25 and 64 years of age (N=104,786). CeVD and haemorrhagic, ischaemic or unspecified stroke were coded with ICD-10 codes I60-I69, I61-I62, I63 and I64, respectively. The analysis included assessment of CeVD deaths distribution and evaluation of age-specific mortality rates in 10-year age groups and age-standardised mortality rates (SMR) in the age group 25-64 years, separately for men and women. Trends in SMRs have been studied in the period 2000-2016. Results. The number of CeVD deaths decreased by 32.8% in men and 48.8% in women. There was a two-fold decline in CeVD mortality: from 59 to 29 male and from 30 to 12 female per 100,000. In addition, a 2-year increase in the median age of CeVD death was observed (Men: 56.4 to 58.4 years, Women: 56.4 to 58.7 years, p<0.001). A statistically significant decline in mortality (per 100,000) was also noticed for haemorrhagic stroke (Men: 18.7 to 10.4; Women: 9.6 to 3.8), ischaemic stroke (Men: 11.8 to 8.4; Women: 4.7 to 3.0) and unspecified stroke (Men: 19.7 to 3.5; Women: 9.1 to 1.3). Conclusions. A substantial decline in premature CeVD mortality was observed in the period 2000-2016. Additionally, the number of deaths that could not be classified as haemorrhagic or ischaemic stroke death decreased significantly. The increasingly widespread use of new post-stroke therapies and their availability make it possible to expect a further decrease in CeVD mortality. However, the necessary actions should be taken to compensate for the disparities in CeVD mortality between men and women.
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