Y.-A. Lee

Identifying fibromyalgia subgroups using cluster analysis: Relationships with clinical variables

Patients with fibromyalgia (FM) exhibit significant clinical heterogeneity, in terms of physical, social and psychological functions, as well as therapeutic responses. Here, we examined FM patients in terms of pain, physical, social and psychological variables to identify clinical subgroups that may be predictive of treatment patterns.

Effect of blood pressure and glycemic control on the plasma cell-free DNA in hemodialysis patients

Background The plasma levels of cell-free DNA (cfDNA) are known to be elevated under inflammatory or apoptotic conditions. Increased cfDNA levels have been reported in hemodialysis (HD) patients. The aim of this study was to investigate the clinical significance of cfDNA in HD patients. Methods A total of 95 patients on HD were enrolled. We measured their predialysis cfDNA levels using real-time EIF2C1 gene sequence amplification and analyzed its association with certain clinical parameters. Results The mean plasma cfDNA level in the HD patients was 3,884 ± 407 GE/mL, and the mean plasma cfDNA level in the control group was 1,420 ± 121 GE/mL (P < 0.05). Diabetic patients showed higher plasma cfDNA levels compared with nondiabetic patients (P < 0.01). Patients with cardiovascular complications also showed higher plasma cfDNA levels compared with those without cardiovascular complication (P < 0.05). In univariable analysis, the cfDNA level was associated with 3-month mean systolic blood pressure (SBP), white blood cell, serum albumin, creatinine (Cr), normalized protein catabolic rate in HD patients. In diabetic patients, it was significantly correlated with SBP, hemoglobin A1c, and serum albumin. In multivariate analysis, SBP was the independent determinant for the cfDNA level. In diabetic patients, cfDNA level was independently associated with hemoglobin A1c and SBP. Conclusions In patients with HD, cfDNA is elevated in diabetic patients and patients with cardiovascular diseases. Uncontrolled hypertension and poor glycemic control are independent determinants for the elevated cfDNA. Our data suggest that cfDNA might be a marker of vascular injury rather than proinflammatory condition in HD patients.

Evaluation of Digital PCR as a Technique for Monitoring Acute Rejection in Kidney Transplantation

Early detection and proper management of kidney rejection are crucial for the long-term health of a transplant recipient. Recipients are normally monitored by serum creatinine measurement and sometimes with graft biopsies. Donor-derived cell-free deoxyribonucleic acid (cfDNA) in the recipients plasma and/or urine may be a better indicator of acute rejection. We evaluated digital PCR (dPCR) as a system for monitoring graft status using single nucleotide polymorphism (SNP)-based detection of donor DNA in plasma or urine. We compared the detection abilities of the QX200, RainDrop, and QuantStudio 3D dPCR systems. The QX200 was the most accurate and sensitive. Plasma and/or urine samples were isolated from 34 kidney recipients at multiple time points after transplantation, and analyzed by dPCR using the QX200. We found that donor DNA was almost undetectable in plasma DNA samples, whereas a high percentage of donor DNA was measured in urine DNA samples, indicating that urine is a good source of cfDNA for patient monitoring. We found that at least 24% of the highly polymorphic SNPs used to identify individuals could also identify donor cfDNA in transplant patient samples. Our results further showed that autosomal, sex-specific, and mitochondrial SNPs were suitable markers for identifying donor cfDNA. Finally, we found that donor-derived cfDNA measurement by dPCR was not sufficient to predict a patients clinical condition. Our results indicate that donor-derived cfDNA is not an accurate predictor of kidney status in kidney transplant patients.

Association between catechol‐O‐methyl transferase gene polymorphisms and fibromyalgia in a Korean population: A case–control study

Although polymorphisms of the catechol‐O‐methyl transferase (COMT) gene have been implicated in altered pain sensitivity, results concerning the association between COMT gene polymorphisms and fibromyalgia (FM) are equivocal. We assessed the associations between COMT single‐nucleotide polymorphisms (SNP) and FM risk and symptom severity.

Both absolute and relative quantification of urinary mRNA are useful for non-invasive diagnosis of acute kidney allograft rejection

Urinary mRNA analysis with three-gene set (18S rRNA, CD3ε, and IP-10) has been suggested as a non-invasive biomarker of acute rejection (AR) in kidney transplant recipients using quantitative real-time PCR (qPCR). Application of droplet digital PCR (ddPCR), which has been suggested to provide higher sensitivity, accuracy, and absolute quantification without standard curves, could be a useful method for the quantifying low concentration of urinary mRNA. We investigated the urinary expression of these three genes in Korean patients with kidney transplantation and also evaluated the usefulness of ddPCR. 90 urine samples were collected at time of allograft biopsy in kidney recipients (n = 67) and from patients with stable renal function more than 10 years (n = 23). Absolute quantification with both PCR system showed significant higher mRNA levels of CD3ε and IP-10 in AR patients compared with stable transplants (STA), but there was no difference in 18S rRNA expression across the patient groups. To evaluate discrimination between AR and STA, ROC curve analyses of CTOT-4 formula yielded area under the curve values of 0.72 (95% CI 0.60–0.83) and 0.77 (95% CI 0.66–0.88) for qPCR and ddPCR, respectively. However, 18S normalization of absolute quantification and relative quantification with 18S showed better discrimination of AR from STA than those of the absolute method. Our data indicate that ddPCR system without standard curve would be useful to determine the absolute quantification of urinary mRNA from kidney transplant recipients. However, comparative method also could be useful and convenient in both qPCR and ddPCR analysis.

AB0048 The role of adiponectin in the production of il-6, il-8, vegf, and mmps in human endothelial cells and osteoblasts: implications for arthritic joints

Background The role of adiponectin in the pathogenesis of rheumatoid arthritis was previously studied in relation to the production of IL-6, IL-8, vascular endothelial growth factor (VEGF), and matrix metalloproteinases (MMPs) in fibroblast-like synoviocytes (FLS). Objectives This study was performed to evaluate the contribution of adiponectin to the production of IL-6, IL-8, VEGF, MMP-1, and MMP-13 in human endothelial cells and osteoblasts in arthritic joints. Methods Cultured human umbilical vascular endothelial cells (HUVEC) and osteoblasts were stimulated with adiponectin (1 or 10 μg/ml) or IL-1β (0.1 ng/ml) in the presence or absence of hypoxia for 24 hrs. The gene expression patterns of culture supernatants and cells were analyzed by ELISA and PCR, respectively. Results Adiponectin significantly stimulated the expression of VEGF, MMP-1, and MMP-13 in osteoblasts but not in HUVEC, while it significantly stimulated the expression of IL-6 and IL-8 in both HUVEC and osteoblasts. The increase of VEGF expression by adiponectin was significantly higher than that by IL-1β stimulation. The expression of IL-6 and IL-8 in adiponectin-stimulated HUVEC was about 10-fold higher than that of IL-1β, but about 2-fold higher in osteoblasts. In addition, IL-8 expression levels in HUVEC were about 7-fold higher than that of osteoblasts. However, IL-6 levels were similar between the two cell types, suggesting that adiponectin may be involved in the production of IL-8 in endothelial cells, which may play an important role in neutrophil recruitment to arthritic joints. Furthermore, the increases observed in gene expression by adiponectin were differentially regulated by hypoxia. Conclusions Adiponectin plays a more important role in the production of mediators driving synovitis and joint destruction in endothelial cells and osteoblasts than that of IL-1β at physiological concentrations. References Arthritis Res Ther 2009, 11(6):R161 Acknowledgements Basic Science Research Program through the National Research Foundation of Korea (NRF) (grant numbers 2010-0024089 and 2011-0009061) Disclosure of Interest None Declared

THU0554 Polymorphisms of Transient Receptor Potential Vanilloid (TRPV) 2 and Trpv3 Gene Polymorphisms Were Associated with fibromyalgia in A Korean Population

Objectives Researchers continue to gather evidence that transient receptor potential vanilloid (TRPV) channels contribute towards pain signaling pathways. However, it is unknown whether polymorphisms of the TRPV gene are associated with fibromyalgia (FM). For the first time, we investigated the association between the polymorphisms of the TRPV2 and TRPV3 genes with FM susceptibility and the severity of the symptoms. Methods A total of 409 patients with FM and 423 controls were enrolled from 10 medical centers that participated in the Korean nationwide FM survey. The alleles and genotypes at three positions [rs3813768 (C>G), rs8121 (C>T), and rs1129235 (C>A)] in the TRPV2 gene, and two positions [rs7216486 (G>A) and rs395357 (C>T)] in the TRPV3 gene, were genotyped. Results The frequencies of the alleles and genotypes of individual TRPV2 and TRPV3 genes were not significantly associated with FM susceptibility. However, the GTA haplotype of TRPV2 showed a defense against FM susceptibility (P=0.035). In addition, polymorphisms of TRPV3 were associated with symptom severity in FM patients. The single nucleotide polymorphism (SNP) rs395357 of TRPV3 was associated with the scores of the Brief Fatigue Inventory (BFI) (P=0.017) in FM patients. Furthermore, haplotypes of TRPV3 were associated with the BFI and the SF-36 mental health summary scores (P=0.036, respectively) Conclusions This study was the first to evaluate the associations of TRPV gene polymorphisms with FM. Our results suggest that certain TRPV2 haplotypes may have a protective role against FM, and also that some genotypes and haplotypes of TRPV3 contribute towards the symptoms of FM. Disclosure of Interest None declared

AB0840 Association between Catechol-O-Methyl Transferase Gene Polymorphisms and fibromyalgia in A Korean Population: A Case-Control Study

Objectives Although polymorphisms of the catechol-O-methyl transferase (COMT) gene have been implicated in altered pain sensitivity, results concerning the association between COMT gene polymorphisms and FM are equivocal. We assessed the associations between COMT single-nucleotide polymorphisms (SNP) and FM risk and symptom severity. Methods In total, 409 FM patients and 423 controls were enrolled. Alleles and genotypes at five positions [rs6269 (A>G), rs4633 (C>T), rs4818 (C>G), rs4680 (C>G) and rs165599 (A>G)] in the COMT gene were genotyped from peripheral blood DNA. Results Alleles and genotypes of the rs4818 COMT gene polymorphism were significantly associated with increased susceptibility to FM. The rs4818 GG genotype was more strongly associated with FM compared to the CC genotype (OR =1.680, 95% CI: 1.057, 2.672, P=0.027). Although allele and genotype frequencies did not differ among groups, the rs4633 CT genotype was not associated with the presence of FM following adjustment for age and sex (OR =0.745; 95% CI: 0.558, 0.995; P=0.046). However, no association was observed between clinical measures and individual COMT SNPs. In haplotype analysis, there was a significant association between ACG haplotype and FM susceptibility sex (OR =2.960, 95% CI: 1.447, 6.056, P=0.003) and the number of tender points (P=0.046). Conclusions This large-scale study suggests that polymorphisms of the COMT gene may be associated with FM risk and pain sensitivity in Korean FM patients. However, our results differed to those of previous studies, suggesting ethnic variation in COMT gene polymorphisms in FM. Disclosure of Interest None declared

AB0787 Evaluation of Longitudinal Association Between Changes in Facet Joints and Administration of Tumor Necrosis Factor Inhibitors in Ankylosing Spondylitis Using CT

Background Ankylosing spondylitis (AS) is a chronic inflammatory arthropathy, which primarily affects the sacroiliac joints and axial skeleton. In AS, involvement of the facet joints results in both back pain, and limitation of spinal mobility. Involvement of facet joint in AS have been reported in several articles. Improvements in spinal mobility in AS after treatment with tumor necrosis factor (TNF) inhibitors have been reported. However, TNF inhibitors were reported to increase or to not inhibit structural damage progression in AS, as assessed using plain radiography. It seems logical that improvement in spinal mobility by TNF inhibitors would result from their effects on facet joint. Objectives To study the effect of TNF inhibitors on progressive facet joint damage in AS patients. Methods AS patients who were treated with TNF inhibitors and had 2 sets of spinal computed tomography (CT) scans, a median of 50.96±5.47 months apart, were included (n=25). Radiographic severity of facet joints was assessed by a new scoring system using CT. Total facet joints from C2-C3 to L5-S1 were scored: erosions 1, subchondral sclerosis 2, partial ankyloses 3, and total ankyloses 4. We compared the longitudinal effects of TNF inhibitors with non-TNF inhibitor treatment by scoring of facet joint of 25 AS patients who were not treated by TNF inhibitors. Results The mean age were 30.9±6.7 years and no significant differences in gender distribution (male, 84% vs. 88%). The only significant difference between two groups was that TNF inhibitor treated group had longer disease duration (years, 4.15±4.02 vs. 2.13±1.73, p=0.001). Significant increases in CT score in the total spine and T-spine were observed in both groups at follow-up time points (Table 1). The rates of increase in CT score of the total spine were similar in both groups, whereas, the rates of increase in CT score of the T-spine was lower in the TNF inhibitor treated group than in the non-TNF group. However, the difference was not statistically significant (p=0.387; Fig. 1).Table 1. CT score of Facet joints at each time point in groups stratified according to TNF inhibitor therapy Initial Follow up (42–66 mo) p-value TNF inhibitor (n=25)  Total spine 33.88±34.37 45.6±41.57 0.004  L-spine 6.36±10.56 7.96±12.68 0.057  T-spine 24.08±22.83 31.44±22.54 0.012  C-spine 3.44±7.52 6.2±11.98 0.460 Non-TNF inhibitor (n=25)  Total spine 18.24±26.58 29.8±33.82 0.000  L-spine 3.88±8.26 4.6±9.45 0.131  T-spine 12.24±17.30 22.8±22.30 0.000  C-spine 2.12±7.01 2.4±7.19 0.465 Values are presented as mean ± SD. Conclusions Treatment with TNF inhibitors did not prevent the radiographic progression of facet joints damage in AS patients References Wilkinson M, Bywaters EG. Clinical features and course of ankylosing spondylitis; as seen in a follow-up of 222 hospital referred cases. Ann Rheum Dis. 1958;17:209-28.2 Machado MA, Barbosa MM, Almeida AM et al. Treatment of ankylosing spondylitis with TNF blockers: a meta-analysis. Rheumatol Int. 2013;33:2199-213. Disclosure of Interest None declared

THU0316 The Symptom Severity in Korean Patients with Fibromyalgia is Associated with Socioeconomic Status, but not with Obesity

Background Fibromyalgia (FM) is a chronic pain disorder characterized by widespread pain, tenderness, and various associated symptoms including sleep disturbances, chronic fatigue, depression, and other somatic symptoms. Several longitudinal and cross-sectional studies in western countries have shown that a high body mass index (BMI) is a strong and independent risk factor for future development of FM and is associated with higher levels of FM symptoms. In addition, obese patients have more physical and emotional impairments compared with nonobese patients. Objectives The purpose of this study was to determine whether obesity and socioeconomic factors influence symptom severity in Korean patients with FM. Methods A total of 343 patients with FM were recruited from outpatient clinics at 11 medical centers across the Republic of Korea. All patients met the ACR 1990 classification criteria for FM. We interviewed these patients using a structured questionnaire that included sociodemographic data, current or past FM symptoms, and current use of relevant medications at the time of enrollment. Tender point counts and scores were assessed by thumb palpation. Patients were asked to complete a Korean version of the Fibromyalgia Impact Questionnaire (FIQ), the Brief Fatigue Inventory (BFI), the SF-36, the Beck Depression Inventory (BDI), the State-Trait Anxiety Inventory (STAI), the Self-Efficacy Scale, and the Social Support Scale. Results Based on an obesity definition of a BMI of ≥25, 76 (22.1%) of the 343 patients were obese. Obese patients were not different from nonobese patients in terms of tender points and scores, FIQ, BFI, SF-36, BDI, STAI, Self-Efficacy, and Social Support scores. After age-, gender-, and symptom duration adjustment by propensity score matching, no significant differences were also found between obese and nonobese patients. However, socioeconomic status such as employment, insurance, and education were significantly associated with symptom severity of FM. The unemployed patients had higher FIQ scores (p=0.011), higher BFI scores (p=0.013), lower physical and mental component SF-36 scores (p=0.012, p=0.005), higher BDI scores (p=0.005), and higher STAI II scores (p=0.041). Lower-income patients had higher FIQ score (p=0.040), lower physical and mental SF-36 scores (p=0.047, p=0.006), higher BDI scores (p<0.000), and lower Self-Efficacy scores (p=0.016). Finally, patients with an education of <12 years had higher tender points (p=0.034), higher BDI scores (p=0.007), and higher STAI II scores (p=0.045). Conclusions Our findings show that, contrary to Western patients, symptom severity in Korean patients with FM is associated with socioeconomic status, but not with obesity. Disclosure of Interest : None declared DOI 10.1136/annrheumdis-2014-eular.2676