## 2 Materials and Methods
### Participants  
The study included Caucasian Southern European individuals recruited from the Neurology Units at Policlinico Umberto I in Rome, Italy. The apparently healthy control group (Normal) consisted of 13 individuals (age 44 \(\pm\) 16 years; 6 females, 7 males), selected from hospital personnel and relatives of patients. All patients with an established diagnosis of MS (\(n\) = 13, age 46 \(\pm\) 13 years; 10 women, 3 men) underwent a detailed neurological examination performed by a senior neurologist expert in demyelinating disorders, confirming their classification as MS.  
MS patients were assessed using the EDSS to quantify overall neurological disability and the FSS to evaluate perceived fatigue levels. The corresponding scores for each scale are shown in Table 1.  
\begin{table}
\begin{tabular}{c c c c c c c c c c c c c} \hline \hline \multirow{2}{*}{ID} & \multirow{2}{*}{**Condition**} & **Age** & \multirow{2}{*}{**Sex**} & \multirow{2}{*}{**Weight**} & \multirow{2}{*}{**Height**} & \multirow{2}{*}{**moll**} & \multirow{2}{*}{**moll**} & \multirow{2}{*}{**EDS**} & \multirow{2}{*}{**FSS**} & \multirow{2}{*}{**DMT**} & \multirow{2}{*}{**ARR**} \\  & & [yrs] & & & [kg] & & & & & & & \\ \hline
**D01** & RRMS & 36 & 1 & 75 & 156 & 31 & 8 & 1 & 18 & Peginterferon beta-1a & 0.2 \\ \hline
**D02** & RRMS & 41 & 0 & 104 & 178 & 33 & 6 & 1 & 11 & Dimethyl fumarate & 0.4 \\ \hline
**D03** & RRMS & 24 & 1 & 80 & 167 & 29 & 4 & 1 & 33 & Ocrelizumab & 0.5 \\ \hline
**D04** & RRMS & 51 & 1 & 51 & 170 & 18 & 24 & 2.5 & 20 & Peginterferon beta-1a & 0.3 \\ \hline
**D05** & RRMS & 19 & 1 & 55 & 168 & 19 & 0.09 & 2 & 21 & Dimethyl fumarate & 1.0 \\ \hline
**D06** & RRMS & 58 & 1 & 48 & 160 & 19 & 14 & 3 & 43 & Figolimod & 0.2 \\ \hline
**D07** & RRMS & 42 & 1 & 48 & 162 & 18 & 4 & 1.5 & 47 & Peginterferon beta-1a & 0.4 \\ \hline
**D08** & RRMS & 55 & 1 & 70 & 170 & 24 & 2 & 2.5 & 28 & Dimethyl fumarate & 0.6 \\ \hline
**D09** & RRMS & 51 & 0 & 85 & 182 & 26 & 20 & 6 & 28 & Ocrelizumab & 0.2 \\ \hline
**D10** & RRMS & 54 & 1 & 95 & 157 & 39 & 7 & 5 & 61 & Dimethyl fumarate & 0.5 \\ \hline
**D11** & RRMS & 56 & 1 & 62 & 167 & 22 & 17 & 6 & 37 & Figolimod & 0.2 \\ \hline
**D12** & RRMS & 57 & 0 & 85 & 181 & 26 & 20 & 3.5 & 32 & Cladribine & 0.2 \\ \hline
**D13** & SPMS & 54 & 1 & 47 & 167 & 17 & 24 & 7.5 & 9 & Ofatumumab & 0.3 \\ \hline
**NT03** & Normal & 55 & 1 & 58 & 168 & 21 & - & - & - & - & - \\ \hline
**NT04** & Normal & 27 & 1 & 54 & 160 & 21 & - & - & - & - & - \\ \hline
**NT05** & Normal & 24 & 1 & 58 & 167 & 21 & - & - & - & - & - \\ \hline
**NT07** & Normal & 36 & 0 & 75 & 174 & 25 & - & - & - & - & - \\ \hline \hline \end{tabular}
\end{table}
Table 1: Demographic and anthropometric characteristics of the study participants, including Expanded Disability Status Scale (EDSS) and Fatigue Severity Scale (FSS) scores for subjects with multiple sclerosis (MS). Control subjects (Normal) did not undergo EDSS or FSS evaluation (indicated by ”-”). Body Mass Index: BMI; Disease Duration: DD; disease-modifying therapy: DMT; Annualized Relapse Rate: ARR. Sex is coded as 0 = male, 1 = female.  
Figure 1: Workflow of the adopted methodology.  
Written informed consent was obtained from all participants prior to their inclusion in the study, which received approval from the local ethics committee (Comitato Etico Lazio 2, protocol number 0167183/2018). All procedures adhered to applicable ethical guidelines and regulatory standards.