Moreover, studies of two large independent ERA cohorts have provided evidence that triple-positive patientsi

Moreover, studies of two large independent ERA cohorts have provided evidence that triple-positive patientsi.e., with RF, ACPA, and anti-carbamylated protein antibodieshad the highest levels of acute phase reactants, suggesting that the amount of inflammation in ERA is proportional to the number of autoantibody-specificities [48]. The concordance of RF+ and ACPA+ in RA is somehow controversial, as observations indicate that ACPA+ patients had disease activity that was similar to, or indeed less than, that of ACPA? patients, both in presence or absence of RF [47]. per se turned out not to predict the response to the therapy, nor did age, sex, body mass index, or smoking status. However, if cases were stratified according to positivity to rheumatoid factor (RF) and anti-citrullinated protein antibody (ACPA) or seronegativity, we observed an opposite association between baseline LINE-1 methylation levels and optimal response to MTX therapy among responders. The best response to MTX therapy was associated with hypermethylated LINE-1 among double-positive ERA cases (= 0.05) and adjusted according to Bonferroni corrections for multiple comparisons when required. The odds ratio (OR) was calculated to evaluate the influence of specific parameters on therapy response. All computations were performed using Microsoft Excel (2016). 3. Results 3.1. Patient Characteristics and Therapy Response Out of 364 recruited CB-1158 patients, 273 were included in the analyses after quality control of data. Details are presented in Table 2. As expected, around KSHV ORF62 antibody three-quarters of the patients were female, with an average age of 59.2 16.0 years, while male patients were on CB-1158 average 650.2 13.1 years old. Overall, CB-1158 approximately half of the cases included in the study were over 60 years old. At the time of ERA diagnosis, the disease length of patients was on average 16.9 13.4 weeks, and the DAS28 score was on average 5.05 1.30. BMI of cases was on average 26.1 5.2 kg/m2, and approximately half of them presented with BMI > 25 kg/m2. Among this group, two-thirds (66.9%) were overweight and one-third (33.1%) were obese (>30 kg/m2). As many as 44.7% of the patients reported being currently exposed to tobacco smoking. Regarding serological characteristics, 56.4% of cases were positive for either RF or ACPA or both (seropositive patients), while 40.3% were positive for both. Table 2 Characteristics of the patients included in the study (= 273). = 0.015), but after adjusting for multiple comparisons it turned out not significant (= 0.057). Among MTX responders, the male patients showed a trend towards association with good response to therapy compared to females (OR = 2.002, 95% CI [1.023C3.917], = 0.042). No association between serological parameters and response to therapy was observed. 3.2. Association with LINE-1 Methylation Level Among the 273 studied cases, the LINE-1 methylation level at baseline was 67.27% on average. As shown in Figure 1, no significant differences were observed among the three response groups. The mean methylation level of non-responders was 67.03 0.71% while it was 67.31 0.22% among MTX responders. Amidst these latter cases, the mean methylation of good responders was 67.24 0.29%, and 67.44 0.36% among moderate responders. Open in a separate window Figure 1 Average LINE-1 methylation level in relation to the response to MTX therapy. Data are presented as the average methylation percentage of five CpG sites. NR, No Response; MR, Moderate Response; GR, Good Response. The correlation of LINE-1 methylation level with response to the therapy was further assessed considering the sex, age class, BMI class, or smoking status of the patients. As presented in Table 3, the LINE-1 methylation level in female or male patients was very similar across all three therapy response groups, and stratification by sex turned out to show no association between LINE-1 methylation and response to therapy. Similar results were obtained after stratification by age class (threshold at age of 60 years) or by exposure to tobacco smoking. Table 3 Average LINE-1 methylation levels with response to the therapy considering sex, age, BMI, and smoking status.

NR 1 MR 2 GR 3 Patients.