Semantic connectivity map obtained with Auto-Cm System.

<p>The figures on the arches of the graph refer to the strength of the association between two adjacent nodes. The range of this value is from 0 to 1.</p

Overview of the folate metabolic pathway, adapted from [18].

<p>Folates require several transport systems to enter the cells, the best characterized being the reduced folate carrier (RFC1). Methylenetetrahydrofolate reductase (MTHFR) reduces 5,10-methylenetetrahydrofolate (5,10-MTHF) to 5-methyltetrahydrofolate (5-MTHF). Subsequently, methionine synthase (MTR) transfers a methyl group from 5-MTHF to homocysteine (Hcy) forming methionine (Met) and tetrahydrofolate (THF). Methionine is then converted to S-adenosylmethionine (SAM) in a reaction catalyzed by methionine adenosyltransferase (MAT). Most of the SAM generated is used in transmethylation reactions, whereby SAM is converted to S-adenosylhomocysteine (SAH) by DNA methyltransferases (DNMTs) that transfer the methyl group to the DNA. Vitamin B12 is a cofactor of MTR, and methionine synthase reductase (MTRR) is required for the maintenance of MTR in its active state. If not converted into methionine, Hcy can be used for the synthesis of glutathione (GSH) in a reaction catalyzed by cystathionine b-synthase (CBS) and other enzymes. Another important function of folate derivatives (THF and dihydrofolate: DHF) is in the de novo synthesis of DNA and RNA precursors (dUMP, dTMP, etc). This pathway is mediated by thymidylate synthase (TYMS), methylenetetrahydrofolate dehydrogenase (MTHFD), and phosphoribosylglycinamide transformylase (GART) enzymes.</p

Demographic characteristics of Myasthenia Gravis patients and Controls.

<p>Gender: no significant difference between MG patients and controls </p><p>Age: no significant difference between MG patients and controls </p><p>Associated autoimmune diseases (AID): Hashimoto’s thyroiditis: 16 cases; Graves-Basedow disease: 14 cases; Type 1 Diabetes: 7 cases; Others: 14 cases.</p

DNMT3B -579G>T allele frequencies in Myasthenia Gravis patients and Controls.

<p>Compared to the control group</p><p>Fisher’s exact test <i>P</i>-value = 0.01; Bonferroni’s corrected <i>P</i>-value = 0.04</p

<i>CDKN2A</i> pyrosequencing.

<p>10 CpG sites analyzed in the CpG Island of the <i>CDKN2A</i> gene. The y axis represents the signal intensity, while the x axis shows the dispensation order. The blue color indicate the % of methylation at each CpG site.</p

Bland-Altman plots.

<p>A) <i>CDKN2A</i> gene methylation; B) <i>APC</i> gene methylation. MS-HRM and pyrosequencing assays are performed on each sample, resulting in 2<i>n</i> data points. Each of the <i>n</i> samples is then represented on the graph by assigning the mean of the two measurements as the abscissa (x-axis) value, and the difference between the two values as the ordinate (y-axis) value.</p

Coefficient of correlation between MS-HRM and pyrosequencing methylation results.

<p>Coefficient of correlation between MS-HRM and pyrosequencing methylation results.</p

Melting curves of <i>APC</i> gene(A): the standards and a sample in duplicate (highlighted); Melting curves of <i>CDKN2A</i> gene (B): the standards and two samples in duplicate (highlighted).

<p>Melting curves of <i>APC</i> gene(A): the standards and a sample in duplicate (highlighted); Melting curves of <i>CDKN2A</i> gene (B): the standards and two samples in duplicate (highlighted).</p

MS-HRM and pyrosequencing conditions and analyzed sequences.

<p>MS-HRM and pyrosequencing conditions and analyzed sequences.</p

Profile of <i>CDKN2A</i> and <i>APC</i> methylation (%) obtained for each sample with the two techniques.

<p>Profile of <i>CDKN2A</i> and <i>APC</i> methylation (%) obtained for each sample with the two techniques.</p