The role of Epstein-Barr virus in multiple sclerosis: from molecular pathophysiology to in vivo imaging

Multiple sclerosis (MS) is a disease of the central nervous system characterized by inflammation, demyelination, and neuronal damage. Environmental and genetic factors are associated with the risk of developing MS, but the exact cause still remains unidentified. Epstein-Barr virus (EBV), vitamin D, and smoking are among the most well-established environmental risk factors in MS. Infectious mononucleosis, which is caused by delayed primary EBV infection, increases the risk of developing MS. EBV may also contribute to MS pathogenesis indirectly by activating silent human endogenous retrovirus-W. The emerging B-cell depleting therapies, particularly anti-CD20 agents such as rituximab, ocrelizumab, as well as the fully human ofatumumab, have shown promising clinical and magnetic resonance imaging benefit. One potential effect of these therapies is the depletion of memory B-cells, the primary reservoir site where EBV latency occurs. In addition, EBV potentially interacts with both genetic and other environmental factors to increase susceptibility and disease severity of MS. This review examines the role of EBV in MS pathophysiology and summarizes the recent clinical and radiological findings, with a focus on B-cells and in vivo imaging. Addressing the potential link between EBV and MS allows the better understanding of MS pathogenesis and helps to identify additional disease biomarkers that may be responsive to B-cell depleting intervention

Coagulation Pathways in Neurological Diseases: Multiple Sclerosis

Significant progress has been made in understanding the complex interactions between the coagulation system and inflammation and autoimmunity. Increased blood-brain-barrier (BBB) permeability, a key event in the pathophysiology of multiple sclerosis (MS), leads to the irruption into the central nervous system of blood components that include virtually all coagulation/hemostasis factors. Besides their cytotoxic deposition and role as a possible trigger of the coagulation cascade, hemostasis components cause inflammatory response and immune activation, sustaining neurodegenerative events in MS. Early studies showing the contribution of altered hemostasis in the complex pathophysiology of MS have been strengthened by recent studies using methodologies that permitted deeper investigation. Fibrin(ogen), an abundant protein in plasma, has been identified as a key contributor to neuroinflammation. Perturbed fibrinolysis was found to be a hallmark of progressive MS with abundant cortical fibrin(ogen) deposition. The immune-modulatory function of the intrinsic coagulation pathway still remains to be elucidated in MS. New molecular details in key hemostasis components participating in MS pathophysiology, and particularly involved in inflammatory and immune responses, could favor the development of novel therapeutic targets to ameliorate the evolution of MS. This review article introduces essential information on coagulation factors, inhibitors, and the fibrinolytic pathway, and highlights key aspects of their involvement in the immune system and inflammatory response. It discusses how hemostasis components are (dys)regulated in MS, and summarizes histopathological post-mortem human brain evidence, as well as cerebrospinal fluid, plasma, and serum studies of hemostasis and fibrinolytic pathways in MS. Studies of disease-modifying treatments as potential modifiers of coagulation factor levels, and case reports of autoimmunity affecting hemostasis in MS are also discussed

Crosstalk between hemostasis inhibitors and cholesterol biomarkers in multiple sclerosis

The individual roles of cholesterol pathway biomarkers (CPB) and hemostasis inhibitors with neuroimaging outcomes were previously investigated in multiple sclerosis (MS). The purpose of this extension study was to investigate potential crosstalk between plasma CPB [total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and apolipoproteins (Apo) ApoA-I, ApoAII, ApoB, ApoC-II and ApoE] and hemostasis inhibitors [heparin cofactor-II (HCII), protein C (PC), protein S (PS), thrombomodulin, ADAMTS13 and PAI-1] in a cohort of 127 MS patients, and 40 healthy individuals (HI). The associations were assessed with regressions. In MS patients, HCII was positively associated with TC, LDL-C, HDL-C and ApoA-I (p=0.028, 0.027, 0.002 and 0.027, respectively) but negatively associated with ApoCII (p=0.018). PC was positively associated with ApoC-II (p=0.001) and ApoB (p=0.016) whereas PS was associated with TC (p=0.024) and ApoE (p=0.003) in MS. The ApoC-II associations were not observed in HI. The negative association between ApoC-II and HCll was an exception amongst other positive associations between CPB and hemostasis inhibitors in MS. CPB do not modulate the PC associations with neurodegeneration in MS

Retinal blood vessel analysis using optical coherence tomography in multiple sclerosis

Background: Both greater retinal neurodegenerative pathology and greater cardiovascular burden have been seen in persons with multiple sclerosis (pwMS).1,2 Moreover, studies have described multiple extracranial and intracranial vasculature changes in pwMS.3 However, only a few studies have examined the retinal vasculature in multiple sclerosis (MS). Objectives: To determine if there are differences in retinal vasculature between pwMS and healthy controls (HCs) and their relationship to peripapillary retinal nerve fiber layer (pRNFL) thickness. Materials and methods: A total of 167 pwMS (113 relapsing-remitting MS (RRMS) and 54 progressive MS (PMS)) and 48 HCs were scanned using optical coherence tomography (OCT). Earlier OCT scans were available in a smaller sample size of 101 pwMS and 35 HCs for additional longitudinal 5-year follow-up analysis. The semiautomated segmentation of the retinal vasculature was performed in a blinded manner on peripapillary scans using the optical coherence tomography segmentation and evaluation GUI (OCTSEG) in MatLab. (Figure 1). Automated segmentation of the pRNFL was performed in the native Heidelberg OCT software. The sum of bilateral measures of total retinal vessel diameter, the total number of retinal vessels and average vessel diameter were calculated. Independent sample t-test and paired t-test were used for cross-sectional and longitudinal analyses, respectively and non-parametric Spearman's test for determining correlations. Results: PwMS had a significantly smaller total vessel diameter (2.5 cm vs 2.7 cm, age-adjusted p=0.017) and numerically fewer number of retinal vessels when compared to HCs (35.1 vs 36.8, age-adjusted p=0.167). No significant differences between the pwRRMS and pwPMS were found. Over the follow-up, pwMS had significant decrease in number of retinal vessels (36.7 vs. 33.0, p<0.001) and significant increase in the average vessel diameter (0.072cm vs. 0.081cm, p<0.001). No longitudinal changes in the HCs were noted. Only in pwMS, lower pRNFL was associated with fewer retinal vessels and total vessel diameter (r=0.191, p=0.018 and r=0.216, p=0.007). Conclusions: PwMS have retinal vasculature that results in smaller and fewer retinal vessels when compared to HCs that were related to reduced pRNFL. Over time, a reduction of retinal vasculature occurred. Future investigations should determine the relevance of retinal vasculature in regards to MS disease outcomes, presence of cardiovascular abnormalities and cerebral/retinal perfusion

Characteristics of an Outpatient Cohort with HBeAg-Negative Chronic Hepatitis B

Introduction: Patients with hepatitis Be antigen negative chronic hepatitis B (HBeAg-negative CHB) and patients inactive carriers (IC) have similar laboratory and serologic characteristics and are not always easy to distinguish. Aim: To characterize hepatitis Be antigen (HBeAg) negative chronic hepatitis B cohort based on their laboratory and virology evaluations at one point of time. Material and Methods: &nbsp;A prospective non-randomized study was conducted on 109 patients with HBeAg negative chronic hepatitis B treated as outpatients at the Clinic for infectious disease and febrile conditions. &nbsp;&nbsp;All patients underwent laboratory and serology testing, quantification of HBV DNA and HBs antigen (qHBsAg). Results: 56 patients were inactive carriers (IC) and 53 patients had HBeAg-negative CHB (AH). The mean values of ALT, HBV DNA and qHBsAg in IC were 29,13 U/L; 727,95 IU/ml and 2753,73 IU/ml respectively. In the AH group the mean values of ALT, HBV DNA and quantitative HBsAg were 50,45 U/L; 7237363,98 IU/ml and 12556,06 IU/ml respectively. The serum value of ALT was more influenced by qHBsAg than HBV DNA in both IC and AH groups (R=0.22 vs R=0.15) (p&gt;0.05). Conclusion: &nbsp;patients with inactive and active HBeAg-negative CHB have similar laboratory and serology profile. It is necessary to combine analysis of ALT, HBV DNA and qHBsAg for better discrimination between patients IC and patient swith HBeAg- negative CHB. Key words: chronic hepatitis B, inactive carriers, ALT, HBeAg, HBV DNA, quantitative HBsAg BACKGROUND: Patients with hepatitis Be antigen-negative chronic hepatitis B (HBeAg-negative CHB), and patients' inactive carriers (IC) have similar laboratory and serologic characteristics and are not always easy to distinguish. AIM: To characterise hepatitis Be antigen (HBeAg) negative chronic hepatitis B cohort based on their laboratory and virology evaluations at one point of time. METHODS: A prospective non-randomized study was conducted on 109 patients with HBeAg negative chronic hepatitis B treated as outpatients at the Clinic for Infectious Diseases and Febrile Conditions. All patients underwent laboratory and serology testing, quantification of HBV DNA and HBs antigen (qHBsAg). RESULTS: A group of 56 patients were inactive carriers (IC), and 53 patients had HBeAg-negative CHB (AH). The mean values of ALT, HBV DNA and qHBsAg in IC were 29.13 U/L; 727.95 IU/ml and 2753.73 IU/ml respectively. In the AH group, the mean values of ALT, HBV DNA and quantitative HBsAg were 50.45 U/L; 7237363.98 IU/ml and 12556.06 IU/ml respectively. The serum value of ALT was more influenced by qHBsAg than HBV DNA in both IC and AH groups (R = 0.22 vs R = 0.15) (p &gt; 0.05). CONCLUSION: patients with inactive and active HBeAg-negative CHB have similar laboratory and serology profile. It is necessary to combine analysis of ALT, HBV DNA and qHBsAg for better discrimination between patient's IC and patient with HBeAg-negative CHB

Cognitive Profiles of Aging in Multiple Sclerosis

BackgroundIncreasingly favorable mortality prognosis in multiple sclerosis (MS) raises questions regarding MS-specific cognitive aging and the presence of comorbidities such as Alzheimer’s disease (AD).ObjectiveTo assess elderly with MS (EwMS) and age-matched healthy controls (HCs) using both MS- and AD-specific psychometrics.MethodsEwMS (n = 104) and 56 HCs were assessed on a broad spectrum of language, visual-spatial processing, memory, processing speed, and executive function tests. Using logistic regression analysis, we examined cognitive performance differences between the EwMS and HC groups. Cognitive impairment (CI) was defined using a -1.5 SD threshold relative to age and education years-matched HCs, in two cognitive domains.ResultsCI was observed in 47.1% of EwMS with differences most often seen on tests emphasizing cognitive processing speed as measured by Symbol Digit Modalities Test (SDMT) (d = 0.9, p &lt; 0.001) and verbal fluency (both category-based d = 0.87, p &lt; 0.001; letter-based d = 0.67, p &lt; 0.001). After adjusting for age, sex and years of education, MS/HC diagnosis was best predicted (R2 = 0.27) by differences in category-based verbal fluency (Wald = 9.935, p = 0.002) and SDMT (Wald = 13.937, p &lt; 0.001).ConclusionThis study confirms the common hallmark of slowed cognitive processing speed in MS among elderly patients. Defective verbal fluency, less often observed in younger cohorts, may represent emerging cognitive pathology due to other etiologies

Phenotypic characteristics and clinical outcome in hospitalized patients with COVID-19 and diabetes

Objectives. The aim is to describe the phenotypic, biological and clinical characteristics of hospitalized patients with COVID-19 and diabetes, and the association with the clinical outcome of the patients. Material and methods. This single-center, retrospective study was conducted on 200 patients. The primary endpoint was death observed within day 7, 14 and beyond day 14 of hospitalization, and secondary objective was to compare the survival group with non-survival group. The variables that demonstrated significant association with primary endpoint were subject to multivariate binary logistic regression analysis. Outcomes. The estimated prevalence was 17.87% of the total COVID-19 hospitalizations during this period (n=1119). The majority of the patients were with diabetes mellitus type 2 with a median age of 67 years and BMI of 27.8 kg/m2. On admission, 156 patients (78%) presented with severe/critical illness. A total of 93 patients (46.5%) met the primary endpoint, with most deaths occurring within day 7 of hospital stay. Non-survival group showed significantly higher levels of leucocytes count, more pronounced lymphopenia, higher CRP, LDH and D-dimer levels. Multivariate analysis identified four independent risk factors associated with death: age OR 1.05 (CI 95% 1.01-1.09), severity of disease at admission OR 0.22 (CI 95, 0.07-0.65), COVID-19 vaccination status OR 3.07 (CI 95%, 1.36-6.91) and LDH levels OR 1.00 (CI 95%,1.002-1.008). Conclusions. Diabetic patients admitted to hospital for COVID-19 infection tend to have high mortality rate. Severity of disease at admission, advanced age, not completed vaccination and increased LDH levels are independent risk factors for lethal outcome, irrespective of diabetes status

Plasma 24-hydroxycholesterol is associated with narrower common carotid artery and greater flow velocities in relapsing multiple sclerosis

Background: Multiple sclerosis (MS) studies suggest greater cardiovascular disease burden and disturbances in the cholesterol pathways1,2 The potential impact of oxidized cholesterol molecules (oxysterols) on MS is emerging (Figure 1).3 Objective: To determine the relationship between multiple oxysterol molecules and atherosclerosis burden in MS patients. Materials and methods: A total of 99 MS patients (61 relapsing-remitting MS (RRMS) and 38 progressive MS (PMS)) patients and 38 healthy controls (HCs) underwent magnetic resonance angiography (MRA) and the cross-sectional area (CSA) of the common carotid artery (CCA) was determined at three different levels before the bifurcation (C7, C6 and C5). Additionally, an echo-color Doppler ultrasound was performed and measures of blood flow velocities were derived. Blood samples acquired at the time of the imaging examinations were analyzed and 24-, 25-, 27-hydroxycholesterol (24HC, 25HC, 27HC) and 7-ketocholesterol (7KC) were quantified in ng/mL. Results: In the MS patients, higher levels of 24HC were significantly associated with smaller CCA CSA measured at all three cervical levels (r=-0.201, p=0.046; r=-0.228, p=0.023, and r=-0.215, p=0.032, for C7, C6 and C5, respectively). These associations were driven by the RRMS group only (r=-0.407, p=0.002 for C7; r=-0.414, p=0.002, for C6; and r=-0.368, p=0.006 for C5). No associations were seen in the HCs. Despite adjusting for the significant age effect (B=0.445, p=0.004), higher 24HC levels were independently associated with smaller CCA CSA (B=-0.20, p=0.045). 24HC was additionally associated with greater time-averaged and peak diastolic CCA velocities. RRMS patients treated with potent anti-inflammatory therapies had lower oxysterol levels (p=0.019). RRMS patients in the lower 24HC quartiles had significantly higher expanded disability status scale (EDSS) scores when compared to RRMS patients in the higher two 24HC quartiles (2.5 (IQR 1.9-3.1) vs 2.0 (1.5-2.5), p=0.038). Conclusions: Greater 24HC levels are associated with smaller CSA CCA and greater flow velocities in RRMS patients. The higher inflammatory activity in RRMS patients may contribute to the production of highly reactive oxysterols and worsen the atherosclerotic burden in the MS population. Potent anti-inflammatory medications can significantly decrease oxysterol levels

Lower cerebral arterial blood flow is associated with greater serum neurofilament light chain levels in multiple sclerosis patients

Background: Hypoperfusion, vascular pathology, and cardiovascular risk factors are associated with disease severity in multiple sclerosis (MS).1,2 In particular, the total cerebral arterial blood flow (CABF), measured as a sum of all arterial flow in the neck, was associated with the cognitive performance of MS patients.3 Objective: To assess relationships between CABF and serum neurofilament light chain (sNfL), as neuronal damage biomarker with good prognostic value and treatment responsiveness.4 If the cerebrovascular changes are an independent pathophysiological factor in MS, a relationship should remain significant after controlling for common MS-based disease measures (i.e., T2 lesion volume and brain volume). Materials and methods: Total CABF was measured in 137 patients (86 clinically isolated syndrome (CIS)/relapsing-remitting (RR) and 51 progressive MS (PMS)) and 48 healthy controls (HCs) using Doppler ultrasound. sNfL was quantitated using a single molecule assay (Simoa). Three point zero T magnetic resonance imaging (MRI) examination allowed quantification of T2 lesion and whole-brain volume (WBV). Multiple linear regression models determined the sNfL associated with CABF after correction for demographic and MRI-derived variables. Results: After adjustment for age, sex and body mass index (BMI), total CABF remained statistically significant and model comparisons showed that CABF explained additional 2.6% of the sNfL variance (β=-0.167, p=0.044). (Table 1) CABF also remained significant in a step-wise regression model (β=0.18, p=0.034) upon the inclusion of T2 lesion burden and WBV effects. The explained sNfL variance improved from 17.4%, 22.7% with the presence of at least 2 CVD variable and 25.8% with both CVD and CABF predictors. Lastly, the disease-modifying therapy was not kept in the final model as an independent predictor of sNfL. Patients in the lowest CABF quartile (CABF≤761 mL/min) had significantly higher sNfL (34.6 pg/mL versus 23.9 pg/mL, adjusted-p=0.042) when compared to the highest quartile (CABF≥1130 mL/min). Conclusions: Lower CABF is associated with increased sNfL in MS patients, highlighting direct and independent relationship between cerebral hypoperfusion and axonal pathology. This relationship remained significant in the CIS/RRMS after adjusting for age, sex, and BMI effects

Editorial of Special Issue &ldquo;Multiple Sclerosis: From Diagnostic Biomarkers to Imaging and Clinical Predictors&rdquo;

Multiple sclerosis (MS) is a chronic, neuroinflammatory and neurodegenerative disease of the central nervous system (CNS) that can present with a plethora of physical and cognitive impairments [...