Sample Of Multiple Sclerosis Patients Research Articles

Recent Advances in Metabolomics and Lipidomics Studies in Human and Animal Models of Multiple Sclerosis.

Multiple sclerosis (MS) is a neurodegenerative and inflammatory disease of the central nervous system (CNS) that leads to a loss of myelin. There are three main types of MS: relapsing-remitting MS (RRMS) and primary and secondary progressive disease (PPMS, SPMS). The differentiation in the pathogenesis of these two latter courses is still unclear. The underlying mechanisms of MS are yet to be elucidated, and the treatment relies on immune-modifying agents. Recently, lipidomics and metabolomics studies using human biofluids, mainly plasma and cerebrospinal fluid (CSF), have suggested an important role of lipids and metabolites in the pathophysiology of MS. In this review, the results from studies on metabolomics and lipidomics analyses performed on biological samples of MS patients and MS-like animal models are presented and analyzed. Based on the collected findings, the biochemical pathways in human and animal cohorts involved were investigated and biological mechanisms and the potential role they have in MS are discussed. Limitations and challenges of metabolomics and lipidomics approaches are presented while concluding that metabolomics and lipidomics may provide a more holistic approach and provide biomarkers for early diagnosis of MS disease.

Peroxynitrite reduces Treg cell expansion and function by mediating IL-2R nitration and aggravates multiple sclerosis pathogenesis

T-helper 17 cells and regulatory T cells (Treg) are critical regulators in the pathogenesis of multiple sclerosis (MS) but the factors affecting Treg/Th17 balance remains largely unknown. Redox balance is crucial to maintaining immune homeostasis and reducing the severity of MS but the underlying mechanisms are unclear yet. Herein, we tested the hypothesis that peroxynitrite, a representative molecule of reactive nitrogen species (RNS), could inhibit peripheral Treg cells, disrupt Treg/Th17 balance and aggravate MS pathology by inducing nitration of interleukin-2 receptor (IL-2R) and down-regulating RAS/JNK-AP-1 signalling pathway. Experimental autoimmune encephalomyelitis (EAE) mouse model and serum samples of MS patients were used in the study. We found that the increases of 3-nitrotyrosine and IL-2R nitration in Treg cells were coincided with disease severity in the active EAE mice. Mechanistically, peroxynitrite-induced IL-2R nitration down-regulated RAS/JNK signalling pathway, subsequently impairing peripheral Treg expansion and function, increasing Teff infiltration into the central nerve system (CNS), aggravating demyelination and neurological deficits in the EAE mice. Those changes were abolished by peroxynitrite decomposition catalyst (PDC) treatment. Furthermore, transplantation of the PDC-treated-autologous Treg cells from donor EAE mice significantly decreased Th17 cells in both axillary lymph nodes and lumbar spinal cord, and ameliorated the neuropathology of the recipient EAE mice. Those results suggest that peroxynitrite could disrupt peripheral Treg/Th17 balance, and aggravate neuroinflammation and neurological deficit in active EAE/MS pathogenesis. The underlying mechanisms are related to induce the nitration of IL-2R and inhibit the RAS/JNK-AP-1 signalling pathway in Treg cells. The study highlights that targeting peroxynitrite-mediated peripheral IL-2R nitration in Treg cells could be a novel therapeutic strategy to restore Treg/Th17 balance and ameliorate MS/EAE pathogenesis. The study provides valuable insights into potential role of peripheral redox balance in maintaining CNS immune homeostasis.

Heteromers Formed by GPR55 and Either Cannabinoid CB1 or CB2 Receptors Are Upregulated in the Prefrontal Cortex of Multiple Sclerosis Patients.

Multiple sclerosis (MS) is an autoimmune, inflammatory, and neurodegenerative disease of the central nervous system for which there is no cure, making it necessary to search for new treatments. The endocannabinoid system (ECS) plays a very important neuromodulatory role in the CNS. In recent years, the formation of heteromers containing cannabinoid receptors and their up/downregulation in some neurodegenerative diseases have been demonstrated. Despite the beneficial effects shown by some phytocannabinoids in MS, the role of the ECS in its pathophysiology is unknown. The main objective of this work was to identify heteromers of cell surface proteins receptive to cannabinoids, namely GPR55, CB1 and CB2 receptors, in brain samples from control subjects and MS patients, as well as determining their cellular localization, using In Situ Proximity Ligation Assays and immunohistochemical techniques. For the first time, CB1R-GPR55 and CB2R-GPR55 heteromers are identified in the prefrontal cortex of the human brain, more in the grey than in the white matter. Remarkably, the number of CB1R-GPR55 and CB2R-GPR55 complexes was found to be increased in MS patient samples. The results obtained open a promising avenue of research on the use of these receptor complexes as potential therapeutic targets for the disease.

Predictors of Disability and Disease Progression Among a Sample of Multiple Sclerosis Patients: an Egyptian Study

Predictors of Disability and Disease Progression Among a Sample of Multiple Sclerosis Patients: an Egyptian Study

Serum Evaluation of Human T-lymphotropic Virus Type 1 and 2 (HTLV-I/II) and Human Immunodeficiency Virus (HIV) in Patients with Multiple Sclerosis (MS)

Background: Multiple Sclerosis (MS) is a multifactorial disease that causes neurological disability. Human retroviruses may have a role in the etiology of several acute and chronic neurological disorders. Objectives: We aimed to investigate the presence of HTLV-I/II and HIV antibodies in the sera of MS patients to assess the relationship between these infections and MS. Methods: This case-control study was conducted on serum samples of MS patients and individuals admitted to the same hospital with no symptoms of MS as the control group. Samples were examined for HIV and HTLV-I/II antibodies using ELISA kits. Results: A total of 79 people, including 38 MS patients and 41 healthy individuals, were evaluated. The frequency of HIV antibodies was 0.0% in both the case and control groups. The HTLV-I/II antibodies were detected in 1 patient in the control group and no patients in the MS group, with no statistically significant difference between the two groups (P: 0.999). Conclusions: This study could not establish a relationship between the presence of HTLV-I/II or HIV antibodies and MS.

The expression profile of HAR1A and HAR1B in the peripheral blood cells of multiple sclerosis patients.

Multiple sclerosis (MS) is a progressive neurodegenerative disease of the central nervous system (CNS) with varying degrees of axonal and neuronal damage. The onset and progression of the disease are influenced by several environmental and genetic variables. Long non-coding RNAs (lncRNAs) have a crucial role in the pathophysiology of MS. Our study aimed to assess the levels of HAR1A and HAR1B lncRNA expression in the blood samples of MS patients and investigate the relationship between these lncRNAs and disease activity. The blood samples of 100 MS patients, including 82 relapsing-remitting (RR), 8 primary progressive (PP), and 10 secondary progressive (SP) MS cases, and 100 healthy controls were collected. Quantitative real-time PCR was used for the evaluation of gene expression. ROC curve analysis was performed to evaluate the diagnostic potential of lncRNA levels. A significant decrease was detected in HAR1A expressions (P < 0.0001), and a moderate increase was also shown in HAR1B of SPMS patients (P value = 0.0189). HAR1A showed different expression levels in patients over forty (P value = 0.034). The expression levels of HAR1A and HAR1B were positively correlated in MS patients (r = 0.2003, P value = 0.0457). In addition, ROC curve results suggested that HAR1A can be introduced as a novel biomarker for MS diagnosis (AUC = 0.776). The low serum level of HAR1A may be a potential molecular biomarker for MS diagnosis; however, no discernible difference was detected in the expression level of HAR1B in the blood samples of MS patients.

The impact of PM2.5, PM10 and NO2 on Covid-19 severity in a sample of patients with multiple sclerosis: A case-control study

BackgroundMany studies investigated the association between air pollution and Covid-19 severity but the only study focusing on patients with Multiple Sclerosis (MS) exclusively evaluated exposure to PM2.5. We aim to study, in a sample of MS patients, the impact of long-term exposure to PM2.5, PM10 and NO2 on Covid-19 severity, described as occurrence of pneumonia. MethodsA 1:2 ratio case-control study was designed, differentiating cases and controls based on Covid-19 pneumonia. Associations between pollutants and outcome were studied using logistic regression. Weighted quantile sum (WQS) logistic regression was used to identify the individual contribution of each pollutant within the mixture; Least Absolute Shrinkage and Selection Operator (LASSO) penalized regression was performed to confirm the variable selection from WQS. All the analyses were adjusted for confounders selected a priori. ResultsOf the 615 eligible patients, 491 patients provided detailed place of exposure and were included in the principal analysis. Higher concentrations of air pollutants were associated with increased odds of developing Covid-19 pneumonia (PM2.5: 3rd vs 1st tercile OR(95% CI)=2.26(1.29;3.96); PM10: 3rd vs 1st tercile OR(95% CI)=2.12(1.22;3.68); NO2: 3rd vs 1st tercile OR(95% CI)=2.12(1.21;3.69)). Pollutants were highly correlated with each other; WQS index was associated to an increased risk of pneumonia (β=0.44; p-value=0.004) and the main contributors to this association were NO2 (41%) and PM2.5 (34%). Consistently, Lasso method selected PM2.5 and NO2. ConclusionsHigher long-term exposure to PM2.5, PM10 and NO2 increased the odds of Covid-19 pneumonia among MS patients and the most dangerous pollutants were NO2 and PM2.5.

Characteristics of cerebral blood flow in an Eastern sample of multiple sclerosis patients: A potential quantitative imaging marker associated with disease severity.

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system that is rare in China. At present, there are no widespread quantitative imaging markers associated with disease severity in MS. Despite several previous studies reporting cerebral blood flow (CBF) changes in MS, no consensus has been reached. In this study, we enrolled 30 Eastern MS patients to investigate CBF changes in different brain regions using the arterial spin labeling technique and their relationship with disease severity. The average CBF in MS patients were higher than those in health controls in various brain regions except cerebellum. The results indicated that MS patients with strongly increased CBF showed worse disease severity, including higher Expanded Disability Status Scale (EDSS) scores and serum neurofilament light chain (sNfL) values than those with mildly increased CBF in the parietal lobes, temporal lobes, basal ganglia, and damaged white matter (DWM). From another perspective, MS patients with worse disease severity (higher EDSS score and sNfL values, longer disease duration) showed increased CBF in parietal lobes, temporal lobes, basal ganglia, normal-appearing white matter (NAWM), and DWM. Correlation analysis showed that there was a strong association among CBF, EDSS score and sNfL. MS patients with strongly increased CBF in various brain regions had more ratio in relapsing phase than patients with mildly increased CBF. And relapsing patients showed significantly higher CBF in some regions (temporal lobes, left basal ganglia, right NAWM) compared to remitting patients. In addition, MS patients with cognitive impairment had higher CBF than those without cognitive impairment in the right parietal lobe and NAWM. However, there were no significant differences in CBF between MS patients with and without other neurologic dysfunctions (e.g., motor impairment, visual disturbance, sensory dysfunction). These findings expand our understanding of CBF in MS and imply that CBF could be a potential quantitative imaging marker associated with disease severity.

The Relationship between Quality of Life and Future Anxiety for a Sample of Multiple Sclerosis Patients based on Selected Variables

Objectives: To identify the relationship between quality of life and future anxiety among a sample of Multiple Sclerosis (MS) patients based on variables of the number of years of injury and marital status.&#x0D; Methodology: The descriptive method was used. The study sample included (141) MS patients who were randomly selected from the total number of patients and members of the MS association in Jordan in 2019/2020. To achieve the objectives of the study, a quality-of-life scale was developed based on the following dimensions: environmental health dimension, family and social dimension, work and study dimension, and religious dimension. Also, the future anxiety scale was developed.&#x0D; Results: The existence of a statistically significant relationship at the level (a = 0.05) between the measure of future anxiety and quality of life among patients with MS. Further, there were no statistically significant differences at the level (a = 0.05) in the value of the correlation coefficient between the anxiety of the future and the quality of life of MS patients refers to the number of years of injury and their social status.&#x0D; Conclusions: Conducting more scientific and psychological studies to shed light on the problems faced by patients with MS in Jordan and all Arab regions.

Improvement in the multiple sclerosis functional composite score by multi-function swing suspension training program

BackgroundPhysical activity has been considered as a promising approach to slow down the disease process in Multiple Sclerosis (MS) patients. The functional impairments of MS have been studied in detail, while evidence of the efficacy of exercise training interventions on the Multiple Sclerosis functional composite (MSFC) score in these patients is limited. The aim of this study was to investigate the improvement in MSFC score by multi-function swing suspension training program (MFSST) in the women with MS. MethodsThe patients were divided into two groups as the intervention and control groups. A total of 47 MS patients completed the MSFC components at baseline and after the intervention: the timed 25-foot walk (T25FW); the 9-hole peg test (9HPT); and paced auditory serial addition test (PASAT). Z scores were created for each test based on control means. ResultsThe MSFC score, 9HPT, T25FW, and PASAT showed a significant increment in comparison with the baseline levels in the four, six, and eight weeks following the first exercise session (all p<0.05). These differences in the control group were not significant. The improvement in the MSFC score and the component Z-scores in the intervention groups was found from the fourth week onwards. ConclusionsThe study findings highlight that the progression of MS disability can be partially compensated by physical exercise. Overall, these results indicate that MFSST can be used as an effective treatment method in patients suffering from MS. Longer (years) exercise studies with larger samples of MS patients, with different MS subtypes, and of different sex, are needed to evaluate the effect of other types of exercise interventions on the MSFC score in MS patients with different disabilities.

Emerging role of neuregulin-1beta1 in pathogenesis and progression of multiple sclerosis

Emerging role of neuregulin-1beta1 in pathogenesis and progression of multiple sclerosis

Spinal cord grey matter atrophy in Multiple Sclerosis clinical practice

PurposeSeveral studies have indicated the relevance of spinal cord grey matter atrophy for Multiple Sclerosis (MS) related disability. This work aimed at evaluating feasibility and clinical relevance of MRI-derived estimations of spinal cord grey matter atrophy in clinical practice. MethodsA convenience sample of MS patients (n=48) and healthy controls (HC, n=11) was scanned using brain sagittal 3D T1w (MPRAGE) and transverse T2w-FLAIR, and transverse single-slice spinal cord 2D heavily T1w (PSIR: phase-sensitive inversion recovery) sequences. An experienced technician with neuroradiological supervision used a semiautomated thresholding method implemented in JIM software on PSIR sequences to obtain cross-sectional area (CSA) of the spinal canal (CSAcanal), whole cord (CSAcord), grey (CSAgm) and white matter (CSAwm) at C2-C3. MPRAGE and T2w-FLAIR sequences were used to obtain brain parenchymal, grey and white matter fractions (BPF, GMF and WMF) using the Statistical Parametric Mapping software. Appropriate statistical tests were used before and after age and sex adjustment. ResultsCSAgm and CSAwm could be obtained in all HC, but only 18 patients (37.5%) due to presence of lesions at C2-C3. Significant univariate associations between EDSS and BPF (rho =−0.376, p=0.015), GMF (rho =−0.287, p=0.069), CSAcanal (rho =−0.310, p=0.049), CSAcord (rho =−0.533 p<0.001) and CSAgm (rho =−0.511, p=0.062) were detected. After age/sex adjustment trends were observed for CSAcanal and CSAcord. ConclusionIn a clinical practice setting, relevance of spinal cord grey matter is confirmed, but cervical cord lesions could greatly hamper its application. Clinical associations with disability have been observed and seem stronger for spinal cord than for brain atrophy measures.

Administration of Maresin-1 ameliorates the physiopathology of experimental autoimmune encephalomyelitis

BackgroundResolution of inflammation is an active and regulated process that leads to the clearance of cell debris and immune cells from the challenged tissue, facilitating the recovery of homeostasis. This physiological response is coordinated by endogenous bioactive lipids known as specialized pro-resolving mediators (SPMs). When resolution fails, inflammation becomes uncontrolled leading chronic inflammation and tissue damage, as occurs in multiple sclerosis (MS).MethodsSPMs and the key biosynthetic enzymes involved in SPM production were analysed by metabololipidomics and qPCR in active brain lesions, serum and peripheral blood mononuclear cells (PBMC) of MS patients as well as in the spinal cord of mice with experimental autoimmune encephalomyelitis (EAE). We also tested the therapeutic actions of the SPM coined Maresin-1 (MaR1) in EAE mice and studied its impact on inflammation by doing luminex and flow cytometry analysis.ResultsWe show that levels of MaR1 and other SPMs were below the limit of detection or not increased in the spinal cord of EAE mice, whereas the production of pro-inflammatory eicosanoids was induced during disease progression. Similarly, we reveal that SPMs were undetected in serum and active brain lesion samples of MS patients, which was linked to impaired expression of the enzymes involved in the biosynthetic pathways of SPMs. We demonstrate that exogenous administration of MaR1 in EAE mice suppressed the protein levels of various pro-inflammatory cytokines and reduced immune cells counts in the spinal cord and blood. MaR1 also decreased the numbers of Th1 cells but increased the accumulation of regulatory T cells and drove macrophage polarization towards an anti-inflammatory phenotype. Importantly, we provide clear evidence that administration of MaR1 in mice with clinical signs of EAE enhanced neurological outcomes and protected from demyelination.ConclusionsThis study reveals that there is an imbalance in the production of SPMs in MS patients and in EAE mice, and that increasing the bioavailability of SPMs, such as MaR1, minimizes inflammation and mediates therapeutic actions. Thus, these data suggest that immunoresolvent therapies, such as MaR1, could be a novel avenue for the treatment of MS.

Information Processing Speed Assessed with Letter Digit Substitution Test in Croatian Sample of Multiple Sclerosis Patients.

Cognitive impairment is a common complaint in people with multiple sclerosis (pwMS). The study objective was to determine the psychometric properties of the letter digit substitution test (LDST) that measures information processing speed and to investigate the impact of relevant predictors of LDST achievement in pwMS. The design was cross-sectional. The study included 87 pwMS and 154 control subjects. The validity of LDST was examined, and a hierarchical regression model was used to explore relevant predictors of LDST success. The LDST had excellent construct validity, as expressed by differences between pwMS and control subjects. Convergent validity of the LDST was supported by a significant moderate correlation with the expanded disability status scale (EDSS) (ρ = −0.36; p < 0.05) and a significantly strong correlation with the multiple sclerosis impact scale (MSIS-29) physical subscale (r = −0.64; p < 0.01). The LDTS score well differentiated the pwMS considering age, education, EDSS, disease duration, comorbidity, and medication therapy. Using the LDST as a criterion variable in pwMS results showed consistent evidence for the age, education, and EDSS impact on LDST performance. The best cut-off score of ≤35 discriminated the control and MS group. LDST proved to be a valid test for assessing information processing speed in pwMS.

Features of the intestinal microbiota composition in multiple sclerosis patients receiving oral disease-modifying therapy

BACKGROUND: Heterogeneous dysbiosis of the intestinal microbiome is a common hallmark of multiple sclerosis. In this pilot study, we compared the level of some gut bacteria in multiple sclerosis patients receiving oral disease-modifying therapy versus untreated.&#x0D; MATERIALS AND METHODS: Subjects were patients with relapsing-remitting or secondary and primary progressive multiple sclerosis. Multiple sclerosis patients were treated by Fingolimod (n = 31), Teriflunomide (n = 21) or were untreated (n = 31). The bacterial levels in stool samples were analyzed by cultivation method and real time PCR.&#x0D; RESULTS: The levels of symbiotic and opportunistic bacterial species in the fecal samples of multiple sclerosis patients receiving disease-modifying therapy were different from those in untreated patients. Also, there was a difference in the spectrum of gastrointestinal tract disorders between these patients. Fingolimod-treated patients showed decreased levels of some bacterial species compared to untreated subjects, including Escherichia coli with regular enzymatic activity, Sutterella wadsworthensis (phylum Proteobacteria), butyrate-producing bacteria Roseburia spp., Faecalibacterium prausnitzii, and Ruminococcus spp. (phylum Firmicutes, class Clostridia). Teriflunomide-treated patients demonstrated decreased levels of Lactobacillus spp. and Enterococcus spp. (phylum Firmicutes, class Bacilli) and Ruminococcus spp. Increased levels of Bifidobacterium spp. were observed in treated and untreated multiple sclerosis patients with higher EDSS scores.&#x0D; CONCLUSIONS: This study shows the negative effect of oral disease-modifying therapy on intestinal microbiota composition and gastrointestinal tract disorders. However, more extensive studies are needed to confirm these preliminary results and develop ways to normalize intestinal dysbiosis in multiple sclerosis patients.

Subgroup Analysis of Individuals with Multiple Sclerosis Showing Cognitive Resilience.

Cognitive dysfunction is known to occur in many individuals with multiple sclerosis (MS). However, little is currently known about MS patients without cognitive impairment, including protective factors associated with intact cognition. The present study considered these issues in a sample of MS patients screened for intact subjective and objective cognitive functioning. Two MS participant groups from a larger sample were derived: i) participants within 1 standard deviation of controls on measures of objective cognition, subjective cognition, and informant-observed subjective cognition [cognitively resilient MS group (MScr)], and ii) those classified as not cognitively resilient (MSncr). Both groups were compared with age- and gender-matched controls. Findings indicated that the MScr group was similar to the MSncr group on most disease and demographic variables, and level of fatigue. The MScr group showed higher estimated baseline intellectual ability and reported less anxiety, subclinical depressive symptoms, and pain interference. MScr participants also showed a trend toward more reported compensatory cognitive strategy use than MSncr participants. The MScr group showed comparable reading recognition and pain symptoms to controls. Our findings provide preliminary information on factors associated with cognitive resilience in MS. Future research should examine resilient individuals with MS to further clarify positive outcomes in this condition.

Nutritional status of multiple sclerosis (MS) patients attending Kasr Alainy MS unit: an exploratory cross-sectional study

BackgroundNutrition was claimed to be a factor in MS causation, course, complications, and management. Several studies were conducted to assess the nutritional status of MS patients; however, few studies were conducted to assess this problem in Egypt. Therefore, the purpose of the current study was to assess the nutritional status of a sample of MS patients.MethodsThe researchers conducted an exploratory cross-sectional study among 76 relapsing-remitting MS (RRMS) patients attending Kasr Alainy Multiple Sclerosis Unit (KAMSU) from October 2018 to January 2019 to assess the nutritional status of a sample of MS patients. Data were collected using a structured interview questionnaire including an inquiry about the socioeconomic status, and nutritional status using anthropometric measurements, patient-generated subjective global assessment (PG-SGA), semi-quantitative food frequency questionnaires (SQFFQ), and hemoglobin level measurement. Assessment of fatigue was done using the Modified Fatigue Impact Scale 5-items version.ResultsThe mean age of the study participants was 30 ± 6 years. The disease duration ranged from 2 to 264 months. Malnutrition was prevalent among 67.1% (27.6 % overweight, 36.8% obese, and 2.6% underweight). Half of the investigated patients were anemic. According to the PG-SGA, more than half of the studied patients (53.9%) were classified as moderately or suspected malnourished. The unhealthy dietary habits such as taking only a few meals, junk food intake and skipping breakfast were observed in considerable proportions of the group. The SQFFQ revealed overconsumption of energy and fat, and less than acceptable consumption of dietary fibers by most of the studied patients.ConclusionsOverweight, obesity, anemia, and unhealthy dietary habits were prevalent among the RRMS patients attending the KAMSU. Nutrition care service is extremely needed for this group of patients.

Small Molecule p53-MDM2 Inhibitors As Potential Regulators of p53-Dependent Genes Expression in Multiple Sclerosis

Transcription factor p53 plays an important role in numerous processes such as apoptosis, cell cycle regulation, metabolism, etc. This protein is believed to regulate apoptosis and neuronal death in various disorders such as multiple sclerosis (MS), Parkinson's and Alzheimer's diseases. In many cases p53 inactivation is caused by overexpression of its negative regulator - E3 ubiquitin ligase MDM2 - that exerts its regulatory functions on p53 by poly-ubiquinating it for proteasomal degradation. Under normal conditions p53 and MDM2 are constituted in an interdependent negative-feedback loop that controls their levels. There are multiple examples in the literature that describe inhibition of MDM2 using small molecules that activate p53 and induce p53-mediated molecular processes, primarily in tumor cells. However, currently the range of potential applications of MDM2 inhibitors is not restricted to cancer, but also includes autoimmune diseases. Therefore, suppression of MDM2 by small molecules is considered as one of the major strategies for activation of p53 and its transcriptional functions, further leading to expression of p53-dependent genes.To evaluate the effect of small molecule p53-MDM2 inhibitors (Nutlin-3a and RG-7112) we have isolated peripheral blood mononuclear cells (PBMCs) of MS patients and healthy donors. First, using colorimetric MTS assay we determined cytotoxicity of the compounds in PBMCs for 0.001-100 uM concentration range and incubation time of 24, 48, 72 hours. It was found that both compounds have only minor effect on cell viability in tested conditions.Finally, we employed Taqman real-time reverse transcription PCR (RT-PCR) to quantitatively analyze gene expression with the use of fluorescence-based detection system BioRad CFX-96 Touch. Freshly isolated cells were treated with both compounds at 5-20 uM concentrantion range. Total RNA was isolated and used to synthesize cDNA via reverse transcription reaction and then real-time PCR was carried out. Relative normalized expression was calculated by normalization to beta-actin, data analyzed using CFX Manager software. All PCR reactions were performed in triplicates. The effect of PBMC treatment was investigated on expression levels of key p53 target genes such as p21, Mdm2, PUMA, GADD45A, BAX, Bcl2 . We observed concentration-dependent expression of p21, Mdm2, Puma, GADD45A and Bax genes. Interestingly, Nutlin-3a demonstrated a more profound effect on these genes compared to RG-7112. Neither Nutlin-3a nor RG-7112 displayed any noticeable effect on the expression of Bcl2 . Importantly, we also noticed that gradual increase in Nutlin-3a concentration leads to higher expression levels of p53-dependent genes in MS patient samples compared to healthy donor samples. In conclusion, we expect that specific MDM2 inhibitors such as Nutlin-3a and RG-7112 can be used for regulation of p53-mediated processes in MS.Further experiments are currently ongoing to examine the effect of p53 activation on expression of various cytokines in stimulated and non-stimulated cells from both MS patients and healthy donors.The study was funded by RFBR research grant 16-34-60213 mol_а_dk. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Illumination of Molecular Pathways in Multiple Sclerosis Lesions and the Immune Mechanism of Matrine Treatment in EAE, a Mouse Model of MS.

The etiology of multiple sclerosis (MS) is not clear, and the treatment of MS presents a great challenge. This study aimed to investigate the pathogenesis and potential therapeutic targets of MS and to define target genes of matrine, a quinolizidine alkaloid component derived from the root of Sophorae flavescens that effectively suppressed experimental autoimmune encephalomyelitis (EAE), an animal model of MS. To this end, the GSE108000 gene data set in the Gene Expression Omnibus Database, which included 7 chronic active MS lesions and 10 control samples of white matter, was analyzed for differentially expressed genes (DEGs). X cell was used to analyze the microenvironmental differences in brain tissue samples of MS patients, including 64 types of immune cells and stromal cells. The biological functions and enriched signaling pathways of DEGs were analyzed by multiple approaches, including GO, KEGG, GSEA, and GSVA. The results by X cell showed significantly increased numbers of immune cell populations in the MS lesions, with decreased erythrocytes, megakaryocytes, adipocytes, keratinocytes, endothelial cells, Th1 cells and Tregs. In GSE108000, there were 637 DEGs, including 428 up-regulated and 209 down-regulated genes. Potential target genes of matrine were then predicted by the network pharmacology method of Traditional Chinese medicine, and 12 key genes were obtained by cross analysis of the target genes of matrine and DEGs in MS lesions. Finally, we confirmed by RT-PCR the predicted expression of these genes in brain tissues of matrine-treated EAE mice. Among these genes, 2 were significantly downregulated and 6 upregulated by matrine treatment, and the significance of this gene regulation was further investigated. In conclusion, our study defined several possible matrine target genes, which can be further elucidated as mechanism(s) of matrine action, and novel targets in the treatment of MS.

The Symbol Digit Modalities Test (SDMT) is sensitive but non-specific in MS: Lexical access speed, memory, and information processing speed independently contribute to SDMT performance

BackgroundThe Symbol Digit Modalities Test (SDMT) is the most sensitive metric of neurocognitive function in multiple sclerosis (MS), and is consistently interpreted as a measure of information processing speed (IPS). ObjectiveTo evaluate the cognitive psychometric profile captured by the SDMT to identify whether different cognitive processes independently underlie performance. MethodsThree samples of MS patients (total n=661; 185 research patients at MS center; 370 clinical patients at MS center; 106 persons with MS from the community) completed objective assessments of neuropsychological function across cognitive domains. Exploratory factor analysis (EFA) was used to derive latent cognitive factor scores, and operationalize cognitive domain composite scores, to understand the unique, shared and redundant contribution of different cognitive domains to SDMT performance using hierarchical multiple regression and commonality analysis. ResultsAcross three independent samples we provide converging strong evidence that the cognitive domains of Memory, IPS and Rapid Automatized Naming (lexical access speed) jointly and uniquely contribute to SDMT performance. ConclusionThe SDMT measures multiple cognitive processes, which likely explains the high degree of sensitivity to cognitive change in MS. Researchers and clinicians should interpret the SDMT as a multifarious measure of general cognition rather than a specific test of IPS.