Quantitative and structural changes of blood platelet cytoskeleton proteins in multiple sclerosis (MS).

Epidemiological studies show that cardiovascular events related to platelet hyperactivity remain the leading causes of death among multiple sclerosis (MS) patients. Quantitative or structural changes of platelet cytoskeleton alter their morphology and function. Here, we demonstrated, for the first time, the structural changes in MS platelets that may be related to their hyperactivity. MS platelets were found to form large aggregates compared to control platelets. In contrast to the control, the images of overactivated, irregularly shaped MS platelets show changes in the cytoskeleton architecture, fragmented microtubule rings. Furthermore, MS platelets have long and numerous pseudopodia rich in actin filaments. We showed that MS platelets and megakaryocytes, overexpress ß1-tubulin and ß-actin mRNAs and proteins and have altered post-translational modification patterns. Moreover, we identified two previously undisclosed mutations in the gene encoding ß1-tubulin in MS. We propose that the demonstrated structural changes of platelet cytoskeleton enhance their ability to adhere, aggregate, and degranulate fueling the risk of adverse cardiovascular events in MS.

Effect of SARS-CoV-2 Infection and BNT162b2 Vaccination on the mRNA Expression of Genes Associated with Angiogenesis.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), discovered in December 2019 in Wuhan, China, caused the coronavirus disease 2019 (COVID-19). Due to the rate of spread of this virus, the World Health Organization, in March 2020, recognised COVID-19 as a worldwide pandemic. The disease is multisystemic with varying degrees of severity. Unfortunately, despite intensive research, the molecular changes caused by SARS-CoV-2 remain unclear. Mechanisms affected by the virus infection include endothelial dysfunction and angiogenesis. Similarly, the vaccines developed so far affect the process of angiogenesis, contributing to the development of undesirable effects on part of the cardiovascular system. The presented research aimed to investigate the impact of the SARS-CoV-2 infection and the Pfizer Comirnaty vaccine (BNT162b2) on the molecular aspect of angiogenesis. We found that convalescents vaccinated with one dose of BNT162b2 were characterised by higher MMP-7 (metalloproteinases 7) expression than non-vaccinated convalescents and healthy volunteers vaccinated with one dose of BNT162b2. Moreover, non-vaccinated convalescents showed increased mRNA expression of ADAMTS1 (ADAM metallopeptidase with thrombospondin type 1 motif 1) compared to healthy volunteers vaccinated with one dose of BNT162b2. In addition, we showed significant sex differences in the expression of MMP-7. In conclusion, the results of our study suggest a significant impact of SARS-CoV-2 infection and vaccination on the course of angiogenesis at the molecular level.

The Role of Supplementation with Natural Compounds in Post-Stroke Patients.

Malnutrition is a serious problem in post-stroke patients. Importantly, it intensifies with hospitalization, and is related to both somatic and psychological reasons, as well as is associated with the insufficient knowledge of people who accompany the patient. Malnutrition is a negative prognostic factor, leading to a reduction in the quality of life. Moreover, this condition significantly extends hospitalization time, increases the frequency of treatment in intensive care units, and negatively affects the effectiveness of rehabilitation. Obtaining growing data on the therapeutic effectiveness of new compounds of natural origin is possible through the use of pharmacodynamic and analytical methods to assess their therapeutic properties. The proper supply of nutrients, as well as compounds of natural origin, is an important element of post-stroke therapy, due to their strong antioxidant, anti-inflammatory, neuroprotective and neuroplasticity enhancing properties. Taking the above into account, in this review we present the current state of knowledge on the benefits of using selected substances of natural origin in patients after cerebral stroke.

The Molecular Aspects of Disturbed Platelet Activation through ADP/P2Y12 Pathway in Multiple Sclerosis.

Epidemiological studies confirm a high risk of ischemic events in secondary-progressive multiple sclerosis (SP MS) patients, directly associated with an increased level of pro-thrombotic activity of platelets. Our work aimed to verify potential molecular abnormalities of the platelet P2Y12 receptor expression and functionality as a cause of an increased risk of thromboembolism observed in the course of MS. We have demonstrated an enhanced platelet reactivity in response to adenosine diphosphate (ADP) in SP MS relative to controls. We have also shown an increased mRNA expression for the P2RY12 gene in both platelets and megakaryocytes, as well as enhanced density of these receptors on the platelet surface. We postulate that one of the reasons for the elevated risk of ischemic events observed in MS may be a genetically or phenotypically reinforced expression of the platelet P2Y12 receptor. In order to analyze the effect of the PAR1 (protease activated receptor type 1) signaling pathway on the expression level of P2Y12, we also analyzed the correlation parameters between P2Y12 expression and the markers of platelet activation in MS induced by selective PAR1 agonist (thrombin receptor activating peptide-6, TRAP-6). Identifying the molecular base responsible for the enlarged pro-thrombotic activity of platelets in SP MS could contribute to the implementation of prevention and targeted treatment, reducing the development of cardiovascular disorders in the course of the disease.

Association of miRNA and mRNA Levels of the Clinical Onset of Multiple Sclerosis Patients.

Multiple sclerosis (MS) is a demyelinating disease characterized by chronic inflammation of the central nervous system, in which many factors can act together to influence disease susceptibility and progression. To date, the exact cause of MS is still unclear, but it is believed to result from an abnormal response of the immune system to one or more myelin antigens that develops in genetically susceptible individuals after their exposure to a, as yet undefined, causal agent. In our study, we assessed the effect of microRNAs on the expression level of neuroprotective proteins, including neurotrophins (BDNF and NT4/5), heat shock proteins (HSP70 and HSP27), and sirtuin (SIRT1) in peripheral blood mononuclear cells in the development of multiple sclerosis. The analysis of dysregulation of miRNA levels and the resulting changes in target mRNA/protein expression levels could contribute to a better understanding of the etiology of multiple sclerosis, as well as new alternative methods of diagnosis and treatment of this disease. The aim of this study was to find a link between neurotrophins (BDNF and NT4), SIRT1, heat shock proteins (HSP27 and HSP27), and miRNAs that are involved in the development of multiple sclerosis. The analysis of the selected miRNAs showed a negative correlation of SIRT1 with miR-132 and miR-34a and of BDNF with 132-3p in PBMCs, which suggests that the miRNAs we selected may regulate the expression level of the studied genes.

Muscle power, contraction velocity and functional performance after stroke.

OBJECTIVE: The goal of this study was to describe muscle function deficit in patients after stroke as well as to define the relationship between maximal muscle power (Pmax ) and optimal shortening velocity (υopt ) with functional efficiency in stroke survivors. MATERIAL AND METHODS: A total of 134 participants were enrolled in the study, including 67 patients after a stroke and 67 volunteers, matched for age and sex (controls). Functional performance was measured with the timed Up and Go test (TUG) and additionally with Rivermead Motor Assessment (RMA) and Barthel Index (BI) in stroke survivors. To assess Pmax and υopt of the knee extensor muscles, a specially equipped Monark cycle ergometer was used. RESULTS: The power generated by stroke survivors was 49.6% that of their peers and muscle contraction velocity was 65.5%. Pmax /kg and υopt were associated with TUG outcomes in both groups. Pmax /kg and υopt were associated with age in the control group, but not in patients after stroke. In multivariate analysis in patients after stroke, TUG was better predicted by Pmax /kg or υopt than by the age. In stroke survivors, both Pmax /kg and υopt were related to the BI and to the RMA total results. Both BI and RMA total were not determined by age. CONCLUSIONS: Muscle power and muscle contraction velocity in patients who have had a stroke within three months have reduced markedly. These factors significantly affect functional performance. Muscle power and optimal shortening velocity are more important determinants of functional status than age in these stroke survivors.

Evaluation of Selected MicroRNAs Expression in Remission Phase of Multiple Sclerosis and Their Potential Link to Cognition, Depression, and Disability.

Accumulating data suggests that miRNAs might play a major role in neuroinflammatory processes. Therefore, our study aimed to first estimate the levels of miR-155, miR-326, and miR-301a in serum of RR-MS patients in the remission phase and then compare the levels of the examined miRNAs at different times after relapse. In this study, 36 RR-MS patients in the remission phase took part. We analyzed two subgroups of RR-MS: one, 1 to 2 months after completing steroid treatment during relapse (post-acute; n = 13) and the other, over 2 years without any relapse (stable; n = 23). Moreover, we made correlations between these biochemical results and clinical parameters of cognitive impairment, depression, and disability. The obtained results presented downregulation of miR-155 and miR-301a (in 94% and 51% samples, respectively) and overexpression of miR-326 (in 72% samples) in RR-MS patients. Moreover, we observed a positive correlation between the relative expression of miRNAs and BDI (Beck Depression Index) for miR-326 (rho = 0.385459, p = 0.022210; Spearman's rank correlation) and miR-301a (rho = 0.435131, p = 0.008991; Spearman rank correlation). We also observed the differences in expression levels between the post-acute and stable phases of RR-MS. The expression levels of miR-301a and miR155 were higher in the post-acute vs. stable phase of remission (2.385 vs. 0.524 and 0.594 vs. 0.147; respectively). Our study, for the first time, presents miRNA expression differences in two stages of remission: post-acute and stable.

Markers of oxidative/nitrative damage of plasma proteins correlated with EDSS and BDI scores in patients with secondary progressive multiple sclerosis.

OBJECTIVES: The objective of the present study was to evaluate oxidative/nitrative stress in the plasma of 50 patients suffering from the secondary progressive course of multiple sclerosis (MS), and to verify its correlation with physical and mental disability as assessed by the Expanded Disability Status Scale (EDSS), and the Beck Depression Inventory (BDI). METHODS: Oxidative and nitrative damage to proteins was determined by the level of carbonyl groups and 3-nitrotyrosine using ELISA test. Based on the reaction with Ellman's reagent, we estimated the concentration of oxidized thiol groups. Additionally, we measured the level of lipid peroxidation. RESULTS: In plasma drawn from MS patients, we observed a significantly higher level of 3-NT (92%; P < 0.0003), carbonyl groups (29%; P < 0.0001) and thiobarbituric acid reactive substances (73%; P < 0.0001), as well as a lower concentration of thiol groups (33%; P < 0.0001), in comparison to healthy subjects. We noted positive correlations between the level of carbonyl groups or 3-NT and both diagnostic parameters, EDSS and BDI. Negative correlations were observed between concentration of -SH groups and EDSS and BDI. CONCLUSION: Our results indicate that impaired red-ox balance can significantly promote neurodegeneration in secondary progressive MS.

Flow cytometric analysis reveals the high levels of platelet activation parameters in circulation of multiple sclerosis patients.

The epidemiological studies confirm an increased risk of cardiovascular disease in multiple sclerosis, especially prothrombotic events directly associated with abnormal platelet activity. The aim of our study was to investigate the level of blood platelet activation in the circulation of patients with chronic phase of multiple sclerosis (SP MS) and their reactivity in response to typical platelets' physiological agonists. We examined 85 SP MS patients diagnosed according to the revised McDonald's criteria and 50 healthy volunteers as a control group. The platelet activation and reactivity were assessed using flow cytometry analysis of the following: P-selectin expression (CD62P), activation of GP IIb/IIIa complex (PAC-1 binding), and formation of platelet microparticles (PMPs) and platelet aggregates (PA) in agonist-stimulated (ADP, collagen) and unstimulated whole blood samples. Furthermore, we measured the level of soluble P-selectin (sP-selectin) in plasma using ELISA method, to evaluate the in vivo level of platelet activation, both in healthy and SP MS subjects. We found a statistically significant increase in P-selectin expression, GP IIb/IIIa activation, and formation of PMPs and PA, as well as in unstimulated and agonist-stimulated (ADP, collagen) platelets in whole blood samples from patients with SP MS in comparison to the control group. We also determined the higher sP-selectin level in plasma of SP MS subjects than in the control group. Based on the obtained results, we might conclude that during the course of SP MS platelets are chronically activated and display hyperreactivity to physiological agonists, such as ADP or collagen.

[Brain stroke - risk of disability and possibilities of improvment in motor and cognitive functioning]. / Udar mózgu ­ ryzyko niepelnosprawnosci oraz mozliwosci poprawy funkcji motorycznych i poznawczych.

Diseases of the central nervous system are the most common cause of mobility and cognitive impairment. Stroke is the leading cause of hospitalization in the Neurological Rehabilitation Departments. Age increases the risk of stroke, therefore monitoring basic medical parameters, including blood pressure especially under age of 65, is a very important part of preventive healthcare. According to data from the National Association of Stroke, in 10% of patients after brain stroke, recovery of motor functions and mental state is almost complete, in 25% impairment is minimal, in 40% the functional and cognitive disability is moderated or significant therefore requires rehabilitation, in 10% in view of impossibility of active rehabilitation patients requires comprehensive nursing care service at home or in special long term care centers. Early mortality after stroke is about 15% of patients. Neurological rehabilitation after the incident of stroke is based on a multidisciplinary, individual approach to the problems arising directly from the consequences of stroke as well as comorbidities and social conditions and welfare in order to enable the best level of functioning at home and in society. The article discusses motor and cognitive impairment, which is important from neurological rehabilitation point of view as well as opportunities for their improvement. In addition, the work includes a description of the major risk factors for stroke, together with chosen predisposing genes statement.

[The effectiveness of comprehensive rehabilitation after a first episode of ischemic stroke]. / Skutecznosc kompleksowej rehabilitacji po pierwszym w zyciu udarze niedokrwiennym mózgu.

UNLABELLED: Ischemic stroke is the most common cause of hospitalization in the Department of Neurological Rehabilitation. Comprehensive rehabilitation is essential for regaining lost functional efficiency. AIM: The aim of study was to evaluate the effectiveness of specific disorder rehabilitation program in 57 patients with first-ever ischemic stroke. MATERIALS AND METHODS: The study included 57 patients (27 women, 30 men) aged from 47 to 89. Patients were admitted for comprehensive rehabilitation, lasted an average of 25 days. The treatment program consisted of exercises aimed at reeducation of posture and gait. In addition, physical treatments were used. Evaluation of the effectiveness of rehabilitation was measured using the Activity Daily Living scale, Modified Rankin Scale, Rivermead Measure Assessment (RMA1-global movements, RMA2-lower limb and trunk, RMA3-upper limb) and the psychological tests - Geriatric Depression Scale (GDS) and Beck Depression Inventory (BDI). RESULTS: As a result of comprehensive rehabilitation treatment, functional status and mental health improvement was observed in relation to the ADL scale by 32% (woman 36%, man 30%), Rankin scale by 22% (woman 22%, man 21%). In the RMA, improvement was observed with the statistical significance of p=0.001 in all of the subscales. The highest rate of improvement affected upper limb function: RMA/3 (41%). In other subscales women have achieved statistically more significant improvement than men (RMA/1-43% versus 25%; RMA/2-41% versus 30%). The results related to the psychological assessment showed statistically significant GDS improvement p<0.001 (<60 years old) and BDI (> 60 years old) in test men (p=0.038). Spearman correlation coefficient showed no relation between mental state and functional improvement (GDS versus ADL; BDI versus ADL). CONCLUSIONS: The 25 days comprehensive rehabilitation program during the subacute stroke phase affects mainly the improvement of upper limb function. Women have achieved better functional improvement in all of the parameters. In addition, it was observed that symptoms of depression were presented in all study group, and the improvement of mental focused primarily on patients after 60 years old.

Poststroke depression as a factor adversely affecting the level of oxidative damage to plasma proteins during a brain stroke.

Poststroke depression, the second most serious psychosomatic complication after brain stroke, leads to delay of the rehabilitation process and is associated with an increased disability and cognitive impairment along with increase in term mortality. Research into the biochemical changes in depression is still insufficiently described. The aim of our study was therefore to evaluate the possible association between plasma protein oxidative/nitrative damages and the development of poststroke depression. We evaluated oxidative/nitrative modifications of specific proteins by measurement of 3-nitrotyrosine and carbonyl groups levels using ELISA test. Additionally, we checked differences in proteins thiol groups by spectrophotometric assay based on reaction between DTNB and thiols. We also evaluated catalase activity in erythrocytes measured as ability to decompose H2O2. Correlation analysis was performed using Spearman's rank. We observed significant (P < 0.001) differences in all oxidative/nitrative stress parameters in brain stroke patients compared to healthy group. Our research shows that oxidative damage of proteins is correlated with the degree of poststroke depression, while nitrative changes do not show any relationship. We demonstrate a positive correlation between the concentration of carbonyl groups and the Geriatric Depression Scale and a negative correlation between the degree of depression and the concentration of -SH groups or catalase activity.