Myeung-Su Lee

PBMC and exosome-derived Hotair is a critical regulator and potent marker for rheumatoid arthritis.

Despite growing importance of long non-coding RNAs (lncRNAs) in normal physiological and disease conditions, our knowledge of RA-related lncRNAs remains limited. Therefore, we aimed to identify lncRNA signatures that have prognostic values in RA. There was a notably high expression level of Hotair in blood mononuclear cells and serum exosome of rheumatoid arthritis (RA) patients, leading the migration of active macrophage. In contrast, markedly lower level of Hotair was detected in differentiated osteoclasts and rheumatoid synoviocytes and enforced expression of Hotair led to significantly decreased levels of MMP-2 and MMP-13. This exploratory study provides novel empirical evidence that Hotair could be one of potential biomarkers for diagnosing RA.

MicroRNA-181b regulates articular chondrocytes differentiation and cartilage integrity.

MicroRNAs are endogenous gene regulators that have been implicated in various developmental and pathological processes. However, the precise identities and functions of the miRNAs involved in cartilage development are not yet well understood. Here, we report that miR-181b regulates chondrocyte differentiation and maintains cartilage integrity, and is thus a potent therapeutic target. MiR-181b was significantly down-regulated during chondrogenic differentiation of TGF-β3-stimulated limb mesenchymal cells, but it was significantly up-regulated in osteoarthritic chondrocytes isolated from the cartilage of osteoarthritis patients. The use of a mimic or an inhibitor to alter miR-181b levels in chondroblasts and articular chondrocytes showed that attenuation of miR-181b reduced MMP-13 expression while inducing type II collagen expression. Furthermore, over-expression of anti-miR-181b significantly reduced the cartilage destruction caused by DMM surgery in mice. In sum, our data suggest that miR-181b is a negative regulator of cartilage development, and that inhibition of miR-181b could be an effective therapeutic strategy for cartilage-related disease.

Use of complementary and alternative medicine by rheumatoid arthritis patients in Korea.

This study measured the prevalence of use of complementary and alternative medicine (CAM) in Korean patients with rheumatoid arthritis (RA). A trained nurse conducted 20-min questionnaire-based interviews at the hospitals when each patient visited as an outpatient. The questionnaire included questions on demographic information, clinical information, and the use of CAM. Of the 153 respondents, 125 (82%) had used CAM; 37% of those who used CAM had started taking CAM products following suggestions from family members and other relatives. In users of CAM, 35% considered that it improved the symptoms of RA, and 14% felt it was effective in achieving psychological relaxation. We categorized treatment into six CAM categories used by the respondents: 84.0% of patients used traditional Oriental medical treatments, 70.4% used plant- and animal-derived over-the-counter health care products, and 13.6% used manual therapies. Most RA patients (64%) would like to try a new type of CAM. About half of the respondents (48%) expected to receive information about CAM from their general practitioner even if most (72%) did not discuss their use of CAM with their doctor. Most of the RA patients in this study used CAM, and half reported beneficial effects. Despite the presence of adverse side effects, patients tended to use CAM without discussing it with their main physicians, suggesting that physicians should be actively involved in the prescription and use of CAM.

Gliotoxin reduces the severity of trinitrobenzene sulfonic acid-induced colitis in mice: Evidence of the connection between heme oxygenase-1 and the nuclear factor-κB pathway in vitro and in vivo

Background: Gliotoxin, a fungal metabolite, has been known to show strong immunosuppressive properties, although its mechanisms are not completely understood. In this report, the authors investigated the mechanism whereby gliotoxin has anti‐inflammatory properties in vitro and in trinitrobenzene sulfonic acid‐induced colitis. Materials and Methods: Body weight, histological scores, and myeloperoxidase activity were evaluated in trinitrobenzene sulfonic acid colitis. Nuclear factor‐&kgr;B (NF‐&kgr;B) p65, tumor necrosis factor‐&agr;, interleukin (IL)‐1&bgr;, IL‐12, and intercellular adhesion molecule‐1 were detected by immunohistochemical staining. IL‐8 secretion was measured by an enzyme‐linked immunosorbent assay. Heme oxygenase‐1 (HO‐1) expression and I‐&kgr;B degradation were analyzed by Western blot. Results: Pretreatment of human epithelial HT‐29 cells with gliotoxin significantly blocked the I‐&kgr;B degradation and NF‐&kgr;B p65 nuclear translocation induced by tumor necrosis factor‐&agr; or IL‐1&bgr;; these were parallel with the inhibition of IL‐8 secretion and intercellular adhesion molecule‐1 expression in the same cells. Interestingly, gliotoxin induced HO‐1 in HT‐29 cells and, in turn, inhibition of HO‐1 activity by a zinc protoporphyrin IX reversed the effects of gliotoxin in terms of I‐&kgr;B degradation, intercellular adhesion molecule‐1 expression, and IL‐8 production. In trinitrobenzene sulfonic acid colitis, gliotoxin administration significantly improved the clinical and histopathological symptoms. Notably, gliotoxin also induced HO‐1 in the colonic mucosa and zinc protoporphyrin IX reversed the protective effects of gliotoxin in trinitrobenzene sulfonic acid colitis. Conclusions: These results demonstrate for the first time that the anti‐inflammatory actions mediated by gliotoxin include HO‐1 induction and the subsequent blockade of NF‐&kgr;B‐dependent signaling pathways in vitro and in vivo. The current results also demonstrate that gliotoxin may be an effective agent for the treatment of diseases characterized by mucosal inflammation.

Iron chelator induces MIP-alpha/CCL20 in human intestinal epithelial cells: implication for triggering mucosal adaptive immunity.

A previous report by this laboratory demonstrated that bacterial iron chelator (siderophore) triggers inflammatory signals, including the production of CXC chemokine IL-8, in human intestinal epithelial cells (IECs). Microarray-based gene expression profiling revealed that iron chelator also induces macrophage inflammatory protein 3 α (MIP-3α)/ CC chemokine-ligand 20 (CCL20). As CCL20 is chemotactic for the cells involved in host adaptive immunity, this suggests that iron chelator may stimulate IECs to have the capacity to link mucosal innate and adaptive immunity. The basal medium from iron chelator deferoxamine (DFO)-treated HT-29 monolayers was as chemotactic as recombinant human CCL20 at equivalent concentrations to attract CCR6+ cells. The increase of CCL20 protein secretion appeared to correspond to that of CCL20 mRNA levels, as determined by real-time quantitative RT-PCR. The efficacy of DFO at inducing CCL20 mRNA was also observed in human PBMCs and in THP-1 cells, but not in human umbilical vein endothelial cells. Interestingly, unlike other proinflammatory cytokines, such as TNF-α and IL-1β, a time-dependent experiment revealed that DFO slowly induces CCL20, suggesting a novel mechanism of action. A pharmacologic study also revealed that multiple signaling pathways are differentially involved in CCL20 production by DFO, while some of those pathways are not involved in TNF-α-induced CCL20 production. Collectively, these results demonstrate that, in addition to some bacterial products known to induce host adaptive immune responses, direct chelation of host iron by infected bacteria may also contribute to the initiation of host adaptive immunity in the intestinal mucosa.

Scrub typhus as a possible aetiology of Guillain–Barré syndrome: two cases

Neurological complications of scrub typhus are reported to be rare. Peripheral nervous system involvement has been reported in only one case. We present two cases of Guillan–Barré syndrome (GBS) associated with scrub typhus. In both cases, the findings of an elevated indirect immunofluorescent antibody titer for Orientia tsutsugamushi and nerve conduction study showing sensory-motor polyneuropathy, have led us to believe that scrub typhus could be one of the antecedent illnesses associated with GBS.

CT imaging biomarker for evaluation of emodin as a potential drug on LPS-mediated osteoporosis mice.

RATIONALE AND OBJECTIVES To identify micro-computed tomography (CT) imaging biomarkers for evaluating the effects of emodin, a potential drug to treat osteoporosis, in the mouse model of lipopolysaccharide (LPS)-mediated osteoporosis. MATERIALS AND METHODS Forty male imprinting control region (ICR) mice with LPS-induced bone resorption were equally divided into four experimental groups: phosphate-buffered saline-treated (control), emodin-treated, LPS-treated, and LPS + emodin-treated groups. Emodin (50 mg/kg) was administered orally on alternate days for 8 days, and LPS (5 mg/kg) was injected intraperitoneally on days 1 and 4. After 8 days, the mice were sacrificed, and micro-CT images of the left proximal femurs were obtained. Three-dimensional images were analyzed by using commercial software to measure the bone volume to total volume fraction (BV/TV), trabecular number (Tb-N), trabecular thickness (Tb-Th), and trabecular separation (Tb-Sp) as CT imaging biomarkers. Histologic analyses of the femurs were performed using hematoxylin and eosin and tartrate-resistant acid phosphatase (TRAP) immunohistochemical staining. RESULTS The LPS + emodin-treated group demonstrated marked suppression of LPS-induced bone resorption compared to the LPS-treated group (BV/TV, 28.84% vs. 40.76%; Tb-N, 2.65 vs. 3.45 mm(-1); Tb-Sp, 300.81 vs. 212.31 μm; Tb-Th, 116.94 vs. 131.25 μm). TRAP immunohistochemical analysis showed fewer osteoclasts per field of tissue in the LPS + emodin-treated group than in the LPS-treated group (27.8 vs. 41.8). The BV/TV, Tb-N, and Tb-Sp data correlated well with the histomorphometric findings. CONCLUSIONS The findings reveal a novel effect of emodin on bone remodeling in the LPS-mediated osteoporotic mouse model. The ex vivo micro-CT imaging is a promising tool for assessing the therapeutic effects of potential drugs on osteoporosis.

Candida Arthritis after Arthroscopic Arthroplasty in a Patient without Predisposing Factors

Because candidiasis is usually associated with immunosuppression, candida arthritis in an immunocompetent patient is rare. The symptoms of candidiasis are similar to bacterial infections, tuberculosis, and autoimmune diseases. In our patient with no predisposing factors, candida arthritis was initially excluded because the probability of occurrence was low. The patient had no leukocytosis, the acid-fast bacteria (AFB) stain was negative, and the autoimmune antibody screen was negative. After Candida parapsilosis was cultured in the synovial fluid, the patient was treated with amphotericin B (0.7 mg/kg/day) and oral fluconazole (400 mg/day). The treatment was successful and there were no side effects of the medications.

AB0400 Open Labeled, Multicenter 24-Week Study To Assess The Efficacy and Safety of Tacrobell® in Active Rheumatoid Arthritis Patients

Background Rheumatoid arthritis (RA) is chronic inflammatory disease characterized by persistent synovitis and structural joint damage with T cell-driven inflammation. Tacrolimus suppress activation of T cells through the inhibition of calcineurin. Objectives We evaluated the efficacy and safety of Tacrobell® (Tacrolimus from Chong Kun Dang Pharma Inc.) in Korean active RA patient who had inadequate response to disease-modifying anti-rheumatic drugs (DMARDs) including Methotrexate (MTX). Methods During the study period from Aug. 2012 to Jun. 2015, in this open labeled, multicenter study, 111 patients were enrolled. Patients were in active disease state with Disease Activity Score28 (DAS28) ≥3.2 despite previously taken at least one conventional DMARD including MTX. Patients had wash out period with DMARDs, except MTX. Patients received Tacrobell® during 24 weeks. The initial dose was 1 mg once daily and was increased to 3mg once daily by 1mg, every 4 weeks. The disease activity was measured by the DAS28-ESR at 4, 8, 16, 24-week after the add on Tacrobell®. Simplified Disease Activity Index (SDAI), Korean Health Assessment Questionnaire (KHAQ)-20, Erythrocyte Sedimentation Rate (ESR), C-Reactive Protein (CRP) and the safety was assessed. Results Data from 97 patients were evaluated in full set analysis. At week-24, EULAR response rate were 83.5% (81 of 97) with improvements from week-16 in 74.2% (72 of 97). Mean DAS28-ESR was continuously decreased of 5.64 at baseline, 4.14 (±1.22, p<0.001) at week-16 and 3.66 (±1.39, p<0.001) at week-24. Efficacy rates according to SDAI were 89.7% (87 of 97) and KHAQ-20 score decreased -2.42 (±4.37, p<0.001) from baseline 7.27 (±4.59) at week-24. Mean ESR was decreased -10.97 (±24.16, p<0.001) at week-16, -14.77 (±24.57, p<0.001) at week-24 from baseline 46.05 (±23.22). Mean CRP was decreased from 2.86 (±7.85, p=0.0578) at baseline to 1.34 (±3.02, p=0.0367) at week-24. The most common adverse events were in gastrointestinal (18 of 108; 16.679%) and respiratory disorder (12 of 108; 16.67%). In serious adverse events (6 of 108, 5.56%), two cases (pneumonia, high glucose level) were related with Tacrobell® and recovered with treatment. At laboratory exam, no abnormal findings with increased BUN or Cr as known common Tacrolimus side effect. Systolic blood pressure increased 2.12 mmHg at week-8. Conclusions This study demonstrated the efficacy of add on Tacrobell® therapy to MTX in patients with active RA. References Tsutomu T., et al (2013) Post-marketing surveillance of the safety and effectiveness of tacrolimus in 3,267 Japanese patients with rheumatoid arthritis Mod Rheumatol.; 24(1):8–16 Takeyuki K., et al (2013) Long-term therapeutic effects and safety of tacrolimus added to methotrexate in patients with rheumatoid arthritis Rheumatol Int.; 33:871–877 Mariko K., et al (2013) Efficacy of adjunct tacrolimus treatment in patients with rheumatoid arthritis with inadequate responses to methotrexate Mod Rheumatol.; 23:788–793 Kawai S., et al. (2011) Efficacy and safety of additional use of tacrolimus in patients with early rheumatoid arthritis with inadequate response to DMARDs-a multicenter, double-blind, parallel-group trial Mod Rheumatol.; 21(5):458–68 Disclosure of Interest None declared

OP0074 Ebselen Is A Potential Anti-Osteoporosis Agent by Suppressing Receptor Activator of Nuclear Factor Kappa-B Ligand-Induced Osteoclast Differentiation In Vitro and Lipopolysaccharide-Induced Inflammatory Bone Destruction In Vivo

Background Ebselen is a non-toxic seleno-organic drug with anti-inflammatory and antioxidant properties that is currently being examined in clinical trials to prevent and treat various diseases, including atherosclerosis, stroke, and cancer. Objectives We investigated the effects of ebselen on RANKL-induced differentiation of osteoclasts and their functions and the underlying molecular mechanisms. Furthermore, we determined the effects of ebselen on LPS-induced bone erosion in vivo. Methods To generate osteoclasts from BMMs, we cultured BMMs for 4 days in the condition of M-CSF and RANKL pretreated with ebselen. The cells were then stained with TRAP solution, rhodamine-conjugated phalloidin for F-actin ring labeling and DAPI solution to detect apoptotic body formation. The change of F-actin ring on mature osteoclasts induced by ebselen was quantified by calculating the ratio of actin ring positive (AR+) osteoclasts versus actin ring negative (AR-) osteoclasts. to detect the formation of apoptotic osteoclasts, we performed TUNEL (TdT-mediated dUTP-biotin nick end-labeling) assay. Primary calvaria osteoblasts and BMCs were co-cultured and were re-seeded in hydroxyapatite-coated plates or dentin slices with or without ebselen. ICR mice were divided into 4 experimental groups comprising 5 mice each: phosphate-buffered saline-treated (control) group, ebselen only-treated group, LPS only-treated group, and LPS- and ebselen-treated group. Ebselen (10 mg/kg) or PBS was administered orally every 8 days, and LPS was injected intraperitoneally on days 1 and 4. Micro-computed tomography (μ-CT) data containing 3D images and bone parameters and histological data were acquired Results Ebselen suppressed the formation of TRAP-positive multinucleated cells in an osteoblast/osteoclast co-culture by regulating the ratio of RANKL/osteoprotegerin secreted by osteoblasts. In addition, ebselen treatment in the early stage of osteoclast differentiation inhibited RANKL-dependent osteoclastogenesis by decreasing the phosphorylation of IκB, PI3K, and Akt in early signaling pathways and by subsequently inducing c-Fos and nuclear factor of activated T-cells c1. Further, ebselen induced apoptosis of osteoclasts in the late stage of osteoclast differentiation. In addition, ebselen treatment suppressed filamentous actin ring formation and bone resorption activity of mature osteoclasts. Reflecting these in vitro effects, administration of ebselen recovered bone loss and its μ-CT parameters in lipopolysaccharide-mediated mouse model. Histological analysis confirmed that ebselen prevented trabecular bone matrix degradation and osteoclast formation in the bone tissues. Finally, it was proved that the anti-osteoclastogenic action of ebselen is achieved through targeting N-methyl-D-aspartate receptor. Conclusions Finally, it was proved that the anti-osteoclastogenic action of ebselen is achieved through targeting N-methyl-D-aspartate (NMDA) receptor. Disclosure of Interest None declared