pinecone/core-2020-05-10-deduplication huggingface dataset:
DNA hypomethylation in rheumatoid arthritis synovial fibroblasts

OBJECTIVE: Rheumatoid arthritis synovial fibroblasts (RASFs) are phenotypically activated and aggressive. We undertook this study to investigate whether the intrinsic activation of RASFs is due to global genomic hypomethylation, an epigenetic modification. METHODS: Global genomic hypomethylation was assessed by immunohistochemistry, flow cytometry, and L1 promoter bisulfite sequencing. The levels of Dnmt1 were determined in synovial tissue and cultured SFs by Western blotting before and after treatment with cytokines and growth factors. Normal SFs were treated for 3 months with a nontoxic dose of the DNA hypomethylation drug 5-azacytidine (5-azaC), and changes in gene expression were revealed using complementary DNA arrays. The phenotypic changes were confirmed by flow cytometry. RESULTS: In situ and in vitro, RASF DNA had fewer 5-methylcytosine and methylated CG sites upstream of an L1 open-reading frame than did DNA of osteoarthritis SFs, and proliferating RASFs were deficient in Dnmt1. Using 5-azaC, we reproduced the activated phenotype of RASFs in normal SFs. One hundred eighty-six genes were up-regulated >2-fold by hypomethylation, with enhanced protein expression. These included growth factors and receptors, extracellular matrix proteins, adhesion molecules, and matrix-degrading enzymes. The hypomethylating milieu induced irreversible phenotypic changes in normal SFs, which resembled those of the activated phenotype of RASFs. CONCLUSION: DNA hypomethylation contributes to the chronicity of RA and could be responsible for the limitation of current therapies
Functional autoantibodies against serpin E2 in rheumatoid arthritis

OBJECTIVE: To search for novel autoantibodies in patients with rheumatoid arthritis (RA) in an effort to better understand the processes of joint destruction in this disease. METHODS: Using a modified SEREX technique and complementary DNA derived from RA synovium, serpin E2 was identified as a novel autoantigen and was analyzed by immunohistochemistry. Levels of anti-serpin E2 autoantibodies in serum and synovial fluid from patients with RA, osteoarthritis (OA), psoriatic arthritis, and ankylosing spondylitis, and/or from healthy individuals were assessed by enzyme-linked immunosorbent assay. Since serpin E2 is an inhibitor of serine proteases, we studied the inhibitory activity of serpin E2 toward its target, urokinase plasminogen activator (uPA), in vitro in the presence of isolated anti-serpin E2 autoantibodies and in vivo using the uPA activity assay. RESULTS: We identified autoantibodies against serpin E2 by the SEREX technique. Serpin E2 was overexpressed in RA synovial tissues as compared with OA synovial tissues. Significantly higher levels of anti-serpin E2 autoantibodies were present in samples of synovial fluid (28%) and serum (22%) from RA patients as compared with OA patients (0 and 6%, respectively) or with healthy individuals (6% of sera). Most importantly, anti-serpin E2 autoantibodies isolated from RA sera reversed the inhibitory activity of serpin E2 by 70%. Furthermore, the levels of anti-serpin E2 autoantibodies correlated with the uPA activity in vivo. CONCLUSION: This study characterizes a functional property of a novel autoantibody in RA. Since anti-serpin E2 autoantibodies interfere with the inhibitory activity of serpin E2 toward serine proteases, they might facilitate the joint destruction in RA
Evaluation of the efficacy and safety of terguride in patients with fibromyalgia syndrome: Results of a twelve-week, multicenter, randomized, double-blind, placebo-controlled, parallel-group study

OBJECTIVE: To assess the efficacy and safety of terguride, a partial dopamine agonist, in patients with fibromyalgia syndrome (FMS). METHODS: In a 12-week, multicenter, double-blind, placebo-controlled, parallel-group study, 99 patients were randomized at a ratio of 2 to 1 to receive terguride or placebo. Over 21 days, the dosage was titrated to a maximum daily dose of 3 mg of terguride or placebo, and this fixed dosage was continued over 9 weeks. The primary efficacy variable was the intensity of pain (100-mm visual analog scale). Secondary efficacy variables included the Fibromyalgia Impact Questionnaire (FIQ) score, the tender point score (TPS), and the Hamilton Depression Scale (HDS) score. During the study, patients were evaluated for the presence of cervical spine stenosis by magnetic resonance imaging (MRI). RESULTS: No significant differences in the change in pain intensity, FIQ score, TPS, or HDS score between baseline and 12 weeks were observed in the terguride group as compared with the placebo group. Cervical spine stenosis was detected in 22% of the patients. Only patients with cervical spine stenosis responded to terguride treatment. FIQ scores improved significantly (per-protocol analysis), and pain intensity, the TPS score, and the HDS score showed a trend toward improvement in the terguride group as compared with the placebo group. Terguride treatment was safe. Only those adverse events already known to be side effects of terguride were observed. Premature termination of the study in patients receiving terguride (26%) occurred predominantly during up-titration and in the absence of comedication for treatment of nausea. CONCLUSION: Terguride treatment did not improve pain, the FIQ score, the TPS, or the HDS score in the total study population. However, a subgroup of patients with cervical spine stenosis seemed to benefit from terguride treatment
Proliferative remodeling of the spatial organization of human superficial chondrocytes distant from focal early osteoarthritis

Objective

Human superficial chondrocytes show distinct spatial organizations, and they commonly aggregate near osteoarthritic (OA) fissures. The aim of this study was to determine whether remodeling or destruction of the spatial chondrocyte organization might occur at a distance from focal (early) lesions in patients with OA.
Methods

Samples of intact cartilage (condyles, patellofemoral groove, and proximal tibia) lying distant from focal lesions of OA in grade 2 joints were compared with location-matched nondegenerative (grade 0–1) cartilage samples. Chondrocyte nuclei were stained with propidium iodide, examined by fluorescence microscopy, and the findings were recorded in a top-down view. Chondrocyte arrangements were tested for randomness or significant grouping via point pattern analyses (Clark and Evans Aggregation Index) and were correlated with the OA grade and the surface cell densities.
Results

In grade 2 cartilage samples, superficial chondrocytes were situated in horizontal patterns, such as strings, clusters, pairs, and singles, comparable to the patterns in nondegenerative cartilage. In intact cartilage samples from grade 2 joints, the spatial organization included a novel pattern, consisting of chondrocytes that were aligned in 2 parallel lines, building double strings. These double strings correlated significantly with an increased number of chondrocytes per group and an increased corresponding superficial zone cell density. They were observed in all grade 2 condyles and some grade 2 tibiae, but never in grade 0–1 cartilage.
Conclusion

This study is the first to identify a distinct spatial reorganization of human superficial chondrocytes in response to distant early OA lesions, suggesting that proliferation had occurred distant from focal early OA lesions. This spatial reorganization may serve to recruit metabolically active units as an attempt to repair focal damage.National Institutes of Health (U.S.) (grant P5O-AR39239)National Institutes of Health (U.S.) (grant R01-AR33236)Deutsche Forschungsgemeinschaft (DFG) (grant RO 2511/1-1)Deutsche Forschungsgemeinschaft (DFG) (grant RO 2511/2-1
Cardiac involvement in Churg-Strauss syndrome

OBJECTIVE: Churg-Strauss syndrome (CSS) is a rare form of systemic vasculitis. Previous studies showing cardiac involvement in CSS patients were limited in the number of patients and were often based solely on clinical manifestations. The aim of the present study was to determine in detail the incidence of cardiac involvement in a large population of ambulatory CSS patients. METHODS: Thirty-two consecutive patients with CSS in remission (mean +/- SD duration of disease between diagnosis and enrollment 6.1 +/- 5.8 years, mean +/- SD age 61 +/- 10 years) who were previously unaware of cardiac involvement were compared with 32 randomly selected age- and sex-matched control subjects, using clinical evaluation, electrocardiography (EKG), echocardiography, and cardiac magnetic resonance imaging (MRI). RESULTS: Detailed cardiac evaluation revealed a 62% prevalence of cardiac involvement in CSS patients compared with 3% in controls (P < 0.001), with clinical symptoms in 26% and 3%, respectively (P = 0.009), EKG abnormalities in 66% and 3%, respectively (P < 0.001), and echocardiographic defects in 50% and 3%, respectively (P < 0.001). Cardiac MRI detected cardiac manifestations in 62% of CSS patients. In the presence of cardiac MRI abnormalities, echocardiography could detect cardiac involvement with a sensitivity of 83% and a specificity of 80%. The absence of symptoms or EKG abnormalities did not exclude cardiac involvement, because abnormalities could still be detected in 38% of these patients at the time of echocardiography or cardiac MRI. CONCLUSION: These results demonstrate a high incidence of cardiac involvement in CSS patients. Systematic cardiac evaluation including detailed imaging is required to properly identify CSS patients with cardiac involvement.status: publishe
ERK-1/2 and p38 in the Regulation of Hypertrophic Changes\ud
of Normal Articular Cartilage Chondrocytes Induced by\ud
Osteoarthritic Subchondral Osteoblasts

Objective. Previous studies have shown the influence\ud
of subchondral bone osteoblasts (SBOs) on phenotypical\ud
changes of articular cartilage chondrocytes\ud
(ACCs) during the development of osteoarthritis (OA).\ud
The molecular mechanisms involved during this process\ud
remain elusive, in particular, the signal transduction\ud
pathways. The aim of this study was to investigate the in\ud
vitro effects of OA SBOs on the phenotypical changes in\ud
normal ACCs and to unveil the potential involvement of\ud
MAPK signaling pathways during this process.\ud
Methods. Normal and arthritic cartilage and bone\ud
samples were collected for isolation of ACCs and SBOs.\ud
Direct and indirect coculture models were applied to\ud
study chondrocyte hypertrophy under the influence of\ud
OA SBOs. MAPKs in the regulation of the cell–cell\ud
interactions were monitored by phosphorylated antibodies\ud
and relevant inhibitors.\ud
Results. OA SBOs led to increased hypertrophic\ud
gene expression and matrix calcification in ACCs by\ud
means of both direct and indirect cell–cell interactions.\ud
In this study, we demonstrated for the first time that OA\ud
SBOs suppressed p38 phosphorylation and induced\ud
ERK-1/2 signal phosphorylation in cocultured ACCs.\ud
The ERK-1/2 pathway inhibitor PD98059 significantly\ud
attenuated the hypertrophic changes induced by conditioned\ud
medium from OA SBOs, and the p38 inhibitor\ud
SB203580 resulted in the up-regulation of hypertrophic\ud
genes in ACCs.\ud
Conclusion. The findings of this study suggest\ud
that the pathologic interaction of OA SBOs and ACCs is\ud
mediated via the activation of ERK-1/2 phosphorylation\ud
and deactivation of p38 phosphorylation, resulting in\ud
hypertrophic differentiation of ACCs
Posttranslational Modification of Collagen Type II : Effects on Antigen-Specific T-cell Tolerance and Autoreactivity in Collagen-Induced Arthritis

Rheumatoid arthritis (RA) is a common chronic inflammatory 
disease affecting peripheral joints in approximately 1% of the world 
population.  Immunization of susceptible strains with CII, leads to 
development of collagen-induced arthritis (CIA), an animal model for 
RA. The aim of this thesis was to investigate mechanisms involved in 
regulation of immunological T-cell tolerance in  CIA by studying 
availability of joint-specific CII for presentation to autoreactive T cells 
in healthy as well as pathological settings.
This work shows that transgenic expression of heterologous CII 
can inhibit expansion and Th1/Th17-skewing of antigen-specific T cells 
upon immunization with heterologous CII. The strength of tolerance 
induction was found to be dependent on the abundance of the selfantigen, the genetic background of the mice, as well as the presence or 
absence of posttranslational modifications on CII. Data indicate that 
joint-specific antigens are readily available for presentation in draining 
lymph nodes to induce immunological tolerance. Furthermore, a defect 
in thymic tolerance induction suggests that certain CII modifications are 
presented differentially depending on the location in the organism (Paper 
IV). 
To obtain these results, established mouse systems were refined by 
generating a T-cell receptor specific antibody (Paper I) or by breeding 
diverse mouse and human transgenes on genetic backgrounds with 
different susceptibilities (Paper II & III). 
Even though it is accepted that T cells play an important role in 
arthritis development, it remains controversial where and how  they
contribute to pathogenic mechanisms after loss of tolerance. In 
summary, this thesis describes  a series of new mouse models that will 
aid to further elucidate the arthritogenic action of T cells in disease 
relevant sites. This will hopefully enlarge the mechanistic framework for 
further investigation of human disease pathogenesis, which might lead to 
new therapeutic strategies to promote self-tolerance in diseased 
individuals
Matriptase is a novel initiator of cartilage matrix degradation in osteoarthritis

Increasing evidence implicates serine proteinases in pathologic tissue turnover. The aim of this study was to assess the role of the transmembrane serine proteinase matriptase in cartilage destruction in osteoarthritis (OA). Serine proteinase gene expression in femoral head cartilage obtained from either patients with hip OA or patients with fracture to the neck of the femur (NOF) was assessed using a low-density array. The effect of matriptase on collagen breakdown was determined in cartilage degradation models, while the effect on matrix metalloproteinase (MMP) expression was analyzed by real-time polymerase chain reaction. ProMMP processing was determined using sodium dodecyl sulfate–polyacrylamide gel electrophoresis/N-terminal sequencing, while its ability to activate proteinase-activated receptor 2 (PAR-2) was determined using a synovial perfusion assay in mice. Matriptase gene expression was significantly elevated in OA cartilage compared with NOF cartilage, and matriptase was immunolocalized to OA chondrocytes. We showed that matriptase activated proMMP-1 and processed proMMP-3 to its fully active form. Exogenous matriptase significantly enhanced cytokine-stimulated cartilage collagenolysis, while matriptase alone caused significant collagenolysis from OA cartilage, which was metalloproteinase-dependent. Matriptase also induced MMP-1, MMP-3, and MMP-13 gene expression. Synovial perfusion data confirmed that matriptase activates PAR-2, and we demonstrated that matriptase-dependent enhancement of collagenolysis from OA cartilage is blocked by PAR-2 inhibition. Elevated matriptase expression in OA and the ability of matriptase to activate selective proMMPs as well as induce collagenase expression make this serine proteinase a key initiator and inducer of cartilage destruction in OA. We propose that the indirect effects of matriptase are mediated by PAR-2, and a more detailed understanding of these mechanisms may highlight important new therapeutic targets for OA treatment
The diagnostic utility of MRI in spondyloarthritis: An international multicentre evaluation of 187 subjects (The MORPHO study)

OBJECTIVE:: To systematically assess the diagnostic utility of MRI to differentiate spondyloarthritis (SpA) patients from patients with non-specific back pain (NSBP) and healthy volunteers using a standardized evaluation of MR images of the sacroiliac joints (SIJ). METHODS:: Five readers blinded to patient and diagnosis independently assessed MRI scans (T1-weighted and STIR sequences) of the SIJ from 187 subjects: 75 patients with AS (symptom duration </=10 years); 27 patients with pre-radiographic inflammatory back pain (IBP) (mean symptom duration 29 months); 26 NSBP and 59 healthy controls </=45 years. Bone marrow edema (BME), fat infiltration, erosion, and ankylosis were recorded according to standardized definitions using an online data entry system. We calculated sensitivity, specificity, positive (LR+) and negative (LR-) likelihood ratios for the diagnosis of SpA based on global assessment of the MRI scans. RESULTS:: Diagnostic utility was high for all 5 readers for both AS (sensitivity 0.90, specificity 0.97, LR+ 44.6) and pre-radiographic IBP patients (sensitivity 0.51, specificity 0.97, LR+ 26.0). Diagnostic utility based solely on detection of BME enhanced sensitivity (67%) for IBP patients but reduced specificity (88%); detection of erosions in addition to BME further enhanced sensitivity (81%) without changing specificity. A single MRI lesion of the SIJ was observed in up to 27% of control individuals. CONCLUSION:: This systematic and standardized evaluation of SIJ in SpA patients showed that MRI has much greater diagnostic utility than documented previously. We present for the first time a data driven definition of a positive MRI for SpA
Extracellular calreticulin is present in the joints of patients with rheumatoid arthritis and inhibits FasL (CD95L)-mediated apoptosis of T cells

addresses: Peninsula College of Medicine and Dentistry, and University of Exeter, Exeter, UK.types: Journal Article; Research Support, Non-U.S. Gov'tThe definitive version is available at www3.interscience.wiley.comThe binding of FasL (CD95L) to its receptor, Fas (CD95), induces apoptosis. Studies have shown that in patients with rheumatoid arthritis (RA), T lymphocytes are resistant to FasL-induced apoptosis in vivo but are susceptible to FasL-induced apoptosis in vitro. Dysfunction in this mechanism may be an important contributor to the pathophysiology of RA. Thus, the present study was undertaken to determine which factors might inhibit FasL-Fas binding in vivo and those that would inhibit apoptosis of T lymphocytes in an in vitro model system
Vulnerability of the superficial zone of immature articular cartilage to compressive injury

Objective

The zonal composition and functioning of adult articular cartilage causes depth-dependent responses to compressive injury. In immature cartilage, shear and compressive moduli as well as collagen and sulfated glycosaminoglycan (sGAG) content also vary with depth. However, there is little understanding of the depth-dependent damage caused by injury. Since injury to immature knee joints most often causes articular cartilage lesions, this study was undertaken to characterize the zonal dependence of biomechanical, biochemical, and matrix-associated changes caused by compressive injury.
Methods

Disks from the superficial and deeper zones of bovine calves were biomechanically characterized. Injury to the disks was achieved by applying a final strain of 50% compression at 100%/second, followed by biomechanical recharacterization. Tissue compaction upon injury as well as sGAG density, sGAG loss, and biosynthesis were measured. Collagen fiber orientation and matrix damage were assessed using histology, diffraction-enhanced x-ray imaging, and texture analysis.
Results

Injured superficial zone disks showed surface disruption, tissue compaction by 20.3 ± 4.3% (mean ± SEM), and immediate biomechanical impairment that was revealed by a mean ± SEM decrease in dynamic stiffness to 7.1 ± 3.3% of the value before injury and equilibrium moduli that were below the level of detection. Tissue areas that appeared intact on histology showed clear textural alterations. Injured deeper zone disks showed collagen crimping but remained undamaged and biomechanically intact. Superficial zone disks did not lose sGAG immediately after injury, but lost 17.8 ± 1.4% of sGAG after 48 hours; deeper zone disks lost only 2.8 ± 0.3% of sGAG content. Biomechanical impairment was associated primarily with structural damage.
Conclusion

The soft superficial zone of immature cartilage is vulnerable to compressive injury, causing superficial matrix disruption, extensive compaction, and textural alteration, which results in immediate loss of biomechanical function. In conjunction with delayed superficial sGAG loss, these changes may predispose the articular surface to further softening and tissue damage, thus increasing the risk of development of secondary osteoarthritis.National Institutes of Health (U.S.) (grant P5O-AR39239)National Institutes of Health (U.S.) (grant R01-AR45779)Deutsche Forschungsgemeinschaft (DFG) (grant RO 2511/1-1)Deutsche Forschungsgemeinschaft (DFG) (grant RO 2511/2-1
Anti-U1 RNP antibodies in cerebrospinal fluid are associated with central neuropsychiatric manifestations in systemic lupus erythematosus and mixed connective tissue disease.

[Objective]To determine the significance of anti–U1 RNP antibodies in the cerebrospinal fluid (CSF) of patients with systemic lupus erythematosus (SLE) or mixed connective tissue disease (MCTD) who have central neuropsychiatric SLE (NPSLE). [Methods]The frequency of antinuclear antibodies including anti–U1 RNP antibodies in the sera and CSF of 24 patients with SLE and 4 patients with MCTD, all of whom had neuropsychiatric syndromes, was determined using an RNA immunoprecipitation assay and an enzyme-linked immunosorbent assay. The frequency of anti–U1 RNP antibodies in the CSF of patients with central NPSLE was examined, and the anti–U1 RNP index ([CSF anti–U1 RNP antibodies/serum anti–U1 RNP antibodies]/[CSF IgG/serum IgG]) was compared with CSF interleukin-6 (IL-6) levels and the albumin quotient (Qalb, an indicator of blood–brain barrier damage). CSF and serum antibodies against U1-70K, U1-A, and U1-C, including autoantigenic regions, were examined, and the U1-70K, U1-A, and U1-C indices as well as the anti–U1 RNP index were calculated. [Results]CSF anti–U1 RNP antibodies with an increased anti–U1 RNP index showed 64.3% sensitivity and 92.9% specificity for central NPSLE. The anti–U1 RNP index did not correlate with CSF IL-6 levels or the Qalb. The anti–U1-70K index was higher than the anti–U1-A and anti–U1-C indices in the CSF of anti–U1 RNP antibody–positive patients with central NPSLE. The major autoantigenic region for CSF anti–U1-70K antibodies appeared to be localized in U1-70K amino acid 141–164 residue within the RNA-binding domain. [Conclusion]The frequency of anti–U1 RNP antibodies in the CSF and the anti–U1 RNP index are useful indicators of central NPSLE in anti–U1 RNP antibody–positive patients. The predominance of anti–U1-70K antibodies in CSF suggests intrathecal anti–U1 RNP antibody production