bluuebunny/crossref_metadata_2025 huggingface dataset:
Juvenile rheumatoid arthritis. Effects of disease activity and recombinant human growth hormone on insulin‐like growth factor 1, insulin‐like growth factor binding proteins 1 and 3, and osteocalcin
Authors: Davies Ursula M. , Jones Jennifer , Reeve Jonathan , Camacho‐Hubner Cecilia , Charlett André , Ansell Barbara M. , Preece Michael A. , Woo Patricia M. M.
Published: February 2007

Objective. To investigate possible mechanisms of growth impairment in children with juvenile rheumatoid arthritis (JRA).

Methods. Eighteen prepubertal children with JRA and growth retardation received recombinant human growth hormone (rHuGH) for 1 year. Growth hormone profiles over 24 hours were obtained before treatment in 12 patients; the levels did not differ from those in “short normal” children. Levels of insulin‐like growth factor 1 (IGF‐1), IGF binding proteins (IGFBPs) 1 and 3, insulin, osteocalcin, and C‐reactive protein (CRP), as well as the erythrocyte sedimentation rate were measured serially. Pretreatment levels were compared with control levels.

Results. In JRA patients, IGF‐1, IGFBP‐3, and osteocalcin levels were significantly lower and insulin levels significantly higher than those in controls, but there was no significant difference in the level of IGFBP‐1. With rHuGH treatment, height velocity and mean levels of IGF‐1, osteocalcin, and insulin increased significantly, while mean levels of IGFBP‐1 fell significantly. Levels of IGFBP‐3 correlated with those of IGF‐1. The height velocity correlated positively with IGF‐1 and osteocalcin, and negatively with IGFBP‐1. Levels of IGFBP‐1 were inversely related to those of insulin and IGF‐1. There was a significant negative correlation between the CRP and height velocity, IGF‐1 level, and osteocalcin level.

Conclusion. IGF‐1 production is impaired in children with active JRA. Treatment with a therapeutic dose of rHuGH can rectify the IGF‐1 deficiency within 4 days, but its effect is adversely influenced by the acute‐phase response, as reflected by an elevated CRP level.
Reference sera for antinuclear antibodies. II. Further definition of antibody specificities in international antinuclear antibody reference sera by immunofluorescence and western blotting
Authors: Smolen Josef S. , Butcher Brian , Fritzler Marvin J. , Gordon Thomas , Hardin John , Kalden Joachim R. , Lahita Robert , Maini Ravinder N. , Reeves Westley , Reichlin Morris , Rothfield Naomi , Takasaki Yoshinari , van Venrooij Walther J. , Tan Eng M.
Published: February 2007

Objective. To define the fine specificity of the 10 reference sera used for determination of antinuclear antibodies (ANA) and ANA subsets which are available from the Arthritis Foundation (AF) and from the Centers for Disease Control and Prevention (CDC).

Methods. AF/CDC sera were assessed by experienced laboratory staff, using indirect immunofluorescence and Western blotting.

Results. The original assignment of fluorescence patterns to 4 reference sera was confirmed, and the fluorescence intensities were determined using reference fluorescent beads. On Western blots, sera AF/CDC2 (anti—SS‐B/La) and AF/CDC7 (anti—SS‐A/Ro) did not detect Ro antigens, sera AF/CDC9 and AF/CDC10 appeared to be monospecific anti—Scl‐70 and anti—Jo‐1 sera, respectively, serum AF/CDC4 (anti—U1 small nuclear RNP) recognized the 70‐kd band, and serum AF/CDC5 recognized the Sm antigen with its multiple bands. Semiquantitative analyses revealed that AF/CDC5, AF/CDC2, and AF/CDC10 were strongly reactive sera, whereas AF/CDC4 and AF/CDC9 were much weaker and should be used at lower dilutions on Western blots.

Conclusion. The AF/CDC ANA reference sera, originally described as reference reagents for indirect immunofluorescence and double immunodiffusion techniques, are also useful for Western blotting. The data presented herein further support the use of these sera for reference purposes.
Breast implants in patients with differentiated and undifferentiated connective tissue disease
Authors: Williams H. James , Weisman Michael H. , Berry Charles C.
Published: February 2007

Objective. To assess the frequency of breast implantation and the relationship of the implants to the onset of symptoms in patients with differentiated and undifferentiated connective tissue disease (CTD).

Methods. We evaluated an inception cohort of patients with differentiated and undifferentiated CTD and symptoms of < 12 months duration when enrolled in 1983–1987. The risk of having breast implants in those patients with early symptoms of CTD was determined in comparison with that in a non‐concurrent control group.

Results. Only 3 of 323 women in the cohort had historical, physical, or chest radiographic evidence of breast implantation. In 1 of the 3 patients, the symptoms of CTD began before the breast implantation. The odds ratio was calculated at 1.15, with a 95% confidence interval ranging from 0.23 to 3.41.

Conclusion. This study showed an absence of significant risk for prior breast implantation surgery in patients with well‐defined or undifferentiated CTD.
Clonal expansion of mycobacterial heat‐shock protein–reactive T lymphocytes in the synovial fluid and blood of rheumatoid arthritis patients
Authors: Celis Linda , Vandevyver Caroline , Geusens Piet , Dequeker Jan , Raus Jef , Zhang Jingwu
Published: February 2007

Objective. To examine the reactivity pattern and T cell receptor (TCR) characteristics of mycobacterial heat‐shock protein 65 (hsp65)‐reactive T cells generated from paired synovial fluid (SF) and peripheral blood (PB) samples obtained from rheumatoid arthritis (RA) patients and from healthy subjects.

Methods. The reactivity pattern of hsp65‐reactive T cell clones generated under limiting‐dilution conditions was analyzed in 3H‐thymidine incorporation assays. The TCR variable regions of these hsp65‐reactive T cells were characterized by polymerase chain reaction with TCR AV‐ and BV‐specific primers and by DNA sequence analysis of the third complementarity‐determining region (CDR3).

Results. The hsp65‐reactive T cells derived both from RA patients and controls preferentially recognized the 1–170 and 303–540 regions of hsp65 and did not cross‐react with human hsp60. The hsp65‐reactive T cell clones derived from RA patients displayed a restricted TCR AV and BV gene usage, which can be attributed to the limited clonal origin(s) of the independent T cell clones, as evidenced by CDR3 sequence analysis. These clonally expanded T cells were found in both PB and SF and in different inflamed joints of RA patients.

Conclusion. Our study suggests that there is in vivo clonal activation and expansion of mycobacterial hsp65‐reactive T cells in patients with RA.
The effects of functional suppression of a membrane‐bound complement regulatory protein, CD59, in the synovial tissue in rats
Authors: Mizuno Masashi , Nishikawa Kazuhiro , Goodfellow Rhian M. , Piddlesden Sara J. , Morgan B. Paul , Matsuo Seiichi , Matsuo Seiichi
Published: February 2007

Objective. To investigate the roles of CD59 in the synovial tissue by functional suppression of CD59.

Methods. Rats treated with cobra venom factor to deplete complement or untreated rats were injected intraarticularly with 0.3 mg of the F(ab′)2 fraction of a monoclonal antibody, 6D1, that inhibits the function of rat CD59. The circumference of knee joints was measured, and histologic changes in the synovium were studied.

Results. Joint swelling, thickening of the synovial tissues, infiltration of inflammatory cells into the synovium, and deposition of membrane attack complex (MAC) on the synovial surface were observed after intraarticular injection of 6D1. The inflammatory reaction reached its peak at 24 hours after injection, and finally subsided to normal within 3 days. It was suggested that functional suppression of CD59 in the synovium induced MAC formation followed by synovitis. Serum complement depletion did not completely suppress this reaction. This indicates that complement existing in the joint space is important for the formation of MAC on the synovial surface and for induction of synovitis.

Conclusion. The membrane‐bound complement regulatory protein, CD59, plays a key role in the protection of joints against MAC‐mediated synovial injury and in maintaining the normal integrity of the joint.
Surgical management of calcinosis cutis universalis in systemic lupus erythematosus
Authors: Cousins Marleen A. M. , Jones Daniel B. , Whyte Michael P. , Monafo William W.
Published: February 2007

Calcinosis cutis is common in several connective tissue diseases but rare in systemic lupus erythematosus (SLE). A 43‐year‐old woman with a 12‐year history of SLE who presented with calcinosis cutis at the time of SLE diagnosis developed a large, ulcerated, draining mass on her left hip. The lesion was excised and skin was grafted, with an excellent early result. The clinical variants and mechanisms of ectopic calcification are discussed, as well as the proposed medical therapies for calcinosis cutis.
Low‐level increases in serum C‐reactive protein are present in early osteoarthritis of the knee and predict progressive disease
Authors: Spector T. D. , Hart D. J. , Nandra D. , Doyle D. V. , Mackillop N. , Gallimore J. R. , Pepys M. B.
Published: February 2007

Objective. To examine the role of low‐grade inflammation in the etiology and progression of early osteoarthritis (OA) of the knee.

Methods. We used a new, high‐sensitivity, automated monoclonal antibody immunoassay for the classic acute‐phase protein, C‐reactive protein (CRP), in serum. Anteroposterior radiographs of the knee with weight bearing were obtained on 845 women (ages 44–67) on entry into a population‐based study of OA in Chingford, North London. In those defined radiologically as “cases,” the knee radiographs were repeated after 4 years.

Results. Levels of CRP were higher in 105 women with knee OA defined radiologically as Kellgren‐Lawrence grade 2+ (median 2.4 mg/liter, interquartile range [IQR] 1.0–5.1), compared with 740 women without OA (median 0.7 mg/liter, IQR 0.3–1.8) (P < 0.001). Median levels of CRP were higher in the 31 women whose disease progressed at least 1 Kellgren‐Lawrence grade (median 2.6 mg/liter, IQR 1.9–4.6), compared with the 39 whose disease did not (median 1.3 mg/liter, IQR 0.6–2.4) (P = 0.006). The significance of these differences persisted after adjustment for age, weight, height, smoking, knee pain, or injury. Classifying disease by the presence of joint space narrowing or osteophytes alone produced similar results.

Conclusion. CRP levels are modestly but significantly increased in women with early knee OA, and higher levels predict those whose disease will progress over 4 years, suggesting that low‐grade inflammation may be a significant aspect of early OA and may be amenable to therapeutic intervention and secondary prevention.
Treatment of early Rheumatoid Arthritis with minocycline or placebo. Results of a Randomized, double‐blind, placebo‐controlled trial
Authors: O'Dell James R. , Haire Claire E. , Palmer William , Drymalski Walter , Wees Steven , Blakely Kent , Churchill Melvin , Eckhoff P. James , Weaver Arthur , Doud Deborah , Erikson Nils , Dietz Fred , Olson Rich , Maloley Pierre , Klassen Lynell W. , Moore Gerald F.
Published: February 2007

Objective. To determine if minocycline is an effective therapy for seropositive rheumatoid arthritis (RA) when used within the first year of disease.

Methods. The Rheumatoid Arthritis Investigational Network enrolled 46 patients with RA of <1 year duration into a 6‐month study of minocycline (100 mg twice daily) versus placebo. All patients were rheumatoid factor positive. The primary end point of the study was successful completion of 6 months of treatment with no drug toxicity while maintaining 50% improvement in composite symptoms of arthritis.

Results. Eighteen of the 46 patients who were enrolled met 50% improvement criteria at 3 months, and maintained at least a 50% improvement for 6 months with no significant drug toxicity. Among them were 15 of the 23 patients (65%) treated with minocycline and 3 of 23 patients (13%) treated with placebo (P < 0.001).

Conclusion. In patients with early seropositive RA, therapy with minocycline is superior to placebo.
Role of nitric oxide in Sjögren's syndrome
Authors: Konttinen Yrjö T. , Platts Louise A. M. , Tuominen Susanne , Eklund Kari K. , Santavirta Nina , Törnwall Jyrki , Sorsa Timo , Hukkanen Mika , Polak Julia M.
Published: February 2007

Objective. To measure levels of salivary nitrite (NO2‐) and to localize nitric oxide synthases (NOS) in the labial salivary glands (LSGs) of patients with Sjögren's syndrome (SS).

Methods. NO2‐ was measured by the Griess reaction. LSGs were analyzed using NADPH‐diaphorase histochemical and immunohistochemical studies to determine the constitutive NOS (neuronal [ncNOS] and endothelial [ecNOS]) and inducible NOS (iNOS) isoforms.

Results. The NO2‐ concentration (mean ± SEM 307 ± 51 μM versus 97 ± 16 μM; P < 0.05) and output (166 ± 46 nmoles/minute versus 37 ± 7 nmoles/minute) were increased in SS patients compared with healthy control subjects. NADPH‐diaphorase was found in some nerve fibers and endothelial cells, and, in SS, was found in myoepithelial, acinar, and ductal epithelial cells, but in only a few inflammatory cells. In SS, ncNOS‐immunoreactive nerve fibers were sparse and ecNOS was found in a minority of the CD31‐positive vascular endothelial cells and acinar cells, whereas iNOS was localized in myoepithelial, acinar, and ductal epithelial cells, often together with tumor necrosis factor α.

Conclusion. Nitrite was found in normal human saliva. NO produced by ncNOS probably acts as a non‐adrenergic, noncholinergic neurotransmitter, whereas that produced by ecNOS exerts a vasodilatory effect. SS patients had increased NO2‐ concentrations, with most of the superfluous salivary NO being produced not by the immigrant inflammatory cells, but rather, by the resident salivary gland cells. NO may contribute to inflammatory damage and acinar cell atrophy in SS.
Reduction in the incidence and severity of collagen‐induced arthritis in DBA/1 mice, using exogenous dehydroepiandrosterone
Authors: Williams Phil J. , Jones Richard H. V. , Rademacher Thomas W.
Published: February 2007

Objective. This study examined the effect of exogenous dehydroepiandrosterone (DHEA) on the onset, incidence, and severity of collagen‐induced arthritis (CIA).

Methods. DHEA was administered subcutaneously prior to arthritis induction in DBA/1 mice, and the severity of the subsequent arthritis was monitored. Serum levels of total IgG and IgG isotype‐specific anti‐murine type II collagen were measured.

Results. Repeated administration of DHEA during arthritis induction delayed the onset and decreased the severity of arthritis in male and female DBA/1 mice. DHEA failed to have an observable effect on established arthritis. IgG isotype autoantibody levels were found to be decreased in the sera of DHEA‐treated mice.

Conclusion. Administration of exogenous DHEA offered protection against the development of CIA. These data support the results of human studies in which low DHEA levels have been identified as a potential risk factor for the development of rheumatoid arthritis. These findings also highlight DHEA as a potential therapy worthy of further investigation.
Direct evidence of high DNA binding activity of transcription factor AP‐1 in rheumatoid arthritis synovium
Authors: Asahara Hiroshi , Fujisawa Koushi , Kobata Tetsuji , Hasunuma Tomoko , Maeda Toshiro , Asanuma Masato , Ogawa Norio , Inoue Hajime , Sumida Takayuki , Nishioka Kusuki
Published: February 2007

Objective. To investigate the possible activation of transcription factor AP‐1 in rheumatoid arthritis (RA) and its involvement in the pathogenesis of RA.

Methods. Synovial tissues and peripheral blood samples were obtained from 25 patients with RA and 5 patients with osteoarthritis (OA) during arthroplasty and synovectomy. The synovial tissue was digested with collagenase and separated into adherent and nonadherent cells by plastic‐adhesion methods. Nuclear extracts obtained from each sample were examined by electrophoretic mobility shift assay to determine the DNA binding activity of AP‐1. The expression of c‐fos and c‐jun messenger RNA (mRNA) was examined by in situ reverse transcription assay.

Results. A markedly high DNA binding activity of AP‐1 was detected in the synovial tissues of RA patients, while virtually no activity or only a little activity was observed in OA patients. Following separation of adherent and nonadherent cells, the AP‐1 activity was mainly detected in adherent cells, which consisted of synovial cells and macrophages. However, the activity was significantly higher in the mononuclear cells infiltrating into RA synovium than in RA peripheral blood mononuclear cells. The high DNA binding activity of AP‐1 in RA correlated with the expression of c‐fos and c‐jun mRNA in situ. Furthermore, AP‐1 binding activity also correlated with disease activity.

Conclusion. In RA synovium, AP‐1 DNA binding activity was constitutively up‐regulated. These findings suggest that AP‐1 may play an important role in the pathogenesis of RA, including synovial hyperplasia and abnormal immune responses.
Recognition of chlamydial antigen by HLA‐B27‐restricted cytotoxic T cells in HLA‐B*2705 transgenic CBA (H‐2k) mice
Authors: Kuon Wolfgang , Lauster Roland , Böttcher Ute , Koroknay Armin , Ulbrecht Matthias , Hartmann Matthias , Grolms Martina , Ugrinovic Sanja , Braun Jürgen , Weiss Elisabeth H. , Sieper Joachim
Published: February 2007

Objective. The association of reactive arthritis (ReA) with HLA‐B27 and the presence of bacterial antigen in joints with ReA suggest that bacterial peptides might be presented by the HLA‐B27 molecule and thus stimulate CD8 T cells. This study was performed to investigate the B27‐restricted cytotoxic T lymphocyte (CTL) response to Chlamydia trachomatis, using the model of HLA‐B27 transgenic mice.

Methods. CBA (H‐2k) mice homozygous for HLA‐B*2705 and human β2‐microglobulin expression were immunized with C trachomatis or with the chlamydial 57‐kd heat‐shock protein (hsp57) coupled to latex beads. Cytotoxicity of lymphocytes from in vivo—primed transgenic mice was tested against C trachomatis—infected targets. Blocking experiments were performed with monoclonal antibodies (MAb) against class I major histocompatibility complex molecules.

Results. A Chlamydia‐specific lysis of both B27‐transfected and nontransfected target cells was observed. This response could be inhibited by anti‐B27 and anti‐H2 MAb. CTL from mice immunized with hsp57 were not able to lyse Chlamydia‐infected target cells, and Chlamydia‐specific CTL could not destroy targets loaded with hsp57.

Conclusion. These results suggest the existence of at least 2 CTL populations in this mouse model: one recognizing peptide of bacteria‐infected cells restricted by HLA‐B*2705 and the other recognizing peptide of bacteria‐infected cells restricted by the murine H‐2Kk molecule. It does not appear that hsp57 is a major target for the CD8 T cell response directed against Chlamydia. This animal model opens the way for identifying bacterial epitopes presented by HLA‐B27, and might thus help to clarify the pathogenesis of B27‐associated diseases.
Endothelial activation in monosodium urate monohydrate crystal‐induced inflammation. In vitro and in vivo studies on the roles of tumor necrosis factor α and interleukin‐1
Authors: Chapman Peter T. , Yarwood Helen , Harrison Andrew A. , Stocker Claire J. , Jamar François , Gundel Robert H. , Peters A. Michael , Haskard Dorian O.
Published: February 2007

Objective. There is relatively little direct evidence for the roles of interleukin‐1 (IL‐1) and tumor necrosis factor α (TNFα) in activating endothelium in vivo. The aim of this study was to use in vitro and in vivo models to investigate the contribution of these cytokines to both E‐selectin expression and the recruitment of polymor‐phonuclear cells (PMN) in monosodium urate monohydrate (MSU) crystal‐induced inflammation.

Methods. MSU crystals were incubated with freshly isolated mononuclear cells, after which the harvested supernatants were tested for their ability to induce E‐selectin expression during coculture with human umbilical vein endothelial cells. Subsequent experiments were performed with the addition of neutralizing anticytokine antibodies/antisera. The role of TNFα was then studied in an MSU crystal‐induced monarthritis model, in the presence or absence of anti‐TNFα (5 mg/kg intravenously). 99mtechnetium (99mTc)‐labeled PMN cells and 111indium (111In)‐labeled anti‐E‐selectin monoclonal antibody (MAb) 1.2B6 were intravenously administered 4 hours after intraarticular injection to quantify PMN recruitment and E‐selectin expression in inflamed joints.

Results. MSU crystals were a potent stimulus for IL‐1 and TNFα production by monocytes in vitro, and these cytokines fully accounted for MSU crystalstimulated, monocyte‐mediated endothelial activation. In the MSU crystal‐induced monarthritis model, TNFα blockade was very effective in suppressing both E‐selectin expression and PMN emigration into the inflamed joints, as judged by gamma‐camera image analysis and postmortem tissue counting following the intravenous injection of 99mTc‐PMN and 111In‐anti‐E‐selectin MAb.

Conclusion. IL‐1 and TNFα appear to be the only factors released by monocytes following incubation with MSU crystals, which induce E‐selectin expression in vitro. Anti‐TNFα is effective in suppressing endothelial activation and PMN recruitment in vivo E‐selectin imaging can be used to assess the endothelial response to therapy and may prove useful for clinical studies.
Transglutaminase activity in aging articular chondrocytes and articular cartilage vesicles
Authors: Rosenthal Ann K. , Derfus Beth A. , Henry Lisa A.
Published: February 2007

Objective. Transglutaminases (TGases) (E.G. 2.3.2.13) catalyze a posttranslational modification of proteins and are associated with biomineralization in growth plate cartilage. Type II TGase participates in the activation of latent transforming growth factor β (TGFß), a crucial factor for both normal cartilage mineralization and the pathologic mineralization that results in calcium pyrophosphate dihydrate (CPPD) crystal formation in aging articular cartilage. To explore a possible association between TGase levels and CPPD crystal formation in mature articular cartilage, TGase activity in articular chondrocytes from old and young pigs and in the articular cartilage vesicle (ACV) fraction of porcine articular cartilage was examined. In addition, the effects of TGase inhibitors on the production of inorganic pyrophosphate (PPi), a process necessary for CPPD crystallogenesis, were determined.

Methods. TGase activity was measured with a radiometric assay in cultured articular chondrocytes from the knee joints of old (3–5 years old) and young (2–6 weeks old) pigs and in the ACVs. PPi levels were measured in chondrocyte‐conditioned media in the presence of TGase inhibitors or control compounds.

Results. Levels of TGase activity in the cytosolic fraction of old chondrocytes were 7‐fold higher than those in identically cultured young chondrocytes. The mean ± SD activity level in the membrane fraction of lysed chondrocytes was 6.0 ± 0.6 units/mg protein in old articular chondrocytes and was undetectable in young chondrocytes. In ACVs, the mean ± SD TGase activity level was 1.23 ± 0.1 units/mg protein. Type II TGase protein was present in chondrocyte cytosol and in ACVs. TGase activity was increased by TGFß to 120% of control values (P < 0.01), and decreased by insulin‐like growth factor 1 to 80% of control values (P < 0.01). TGase inhibitors blocked media accumulation of PPi, an essential precursor of CPPD crystal formation, and a sensitive marker of TGFß effect.

Conclusion. These data suggest a potential link between TGase activity and processes of pathologic biomineralization that result in CPPD crystal formation in aging articular cartilage.
In vivo suppression of early experimental osteoarthritis by interleukin‐1 receptor antagonist using gene therapy
Authors: Pelletier Jean‐Pierre , Caron John P. , Evans Christopher , Robbins Paul D. , Georgescu Helga I. , Jovanovic Dragan , Fernandes Julio C. , Martel‐Pelletier Johanne
Published: February 2007

Objective. This study explored the therapeutic effect of interleukin‐1 receptor antagonist (IL‐1Ra), administered by gene transfer, on the progression of osteoarthritic (OA) lesions in an experimental dog model.

Methods. Seventeen mature mongrel dogs were divided into 3 groups. Group 1 (n = 7) had an anterior cruciate ligament (ACL) section of the right knee through a stab wound incision. Groups 2 and 3 (n = 5 per group), had an ACL section of the right knee and partial synovectomy of the left knee. Each dog's synovium was subjected to enzymatic digestion, and the synovial fibroblasts were propagated in monolayer culture. Synovial cells from each dog were transduced in vitro using the retrovirus MFG with either the Escherichia coli β‐galactosidase (lac Z) gene (group 2) or the human IL‐1Ra gene (group 3). Two days after surgery, the dogs received intraarticular injections as follows: group 1 phosphate buffered saline (PBS) (2 ml); group 2 autologous cells (60 × 106 cells/2 ml of PBS) transduced with the lac Z gene; group 3 autologous cells transduced with the IL‐1Ra gene. Synovial fluid was aspirated at 2 weeks and 4 weeks. All dogs were euthanized at 4 weeks postsurgery. The right knees were dissected, and lesions were scored for macroscopic and microscopic changes. Synovial explants were dissected and representative specimens were used for histology or were cultured for 48 hours. The levels of IL‐1Ra in synovial fluid and synovial explant conditioned medium were measured by specific enzyme‐linked immunosorbent assay.

Results. The level of IL‐1Ra in synovial fluid of group 3 was 202.8 ± 131.5 ng/ml (mean ± SEM) at 2 weeks and 2.8 ± 2.2 ng/ml at 4 weeks after surgery. Membrane explants isolated from dogs that received synovial cells transduced with the IL‐1Ra gene (group 3) actively produced IL‐1Ra (4.0 ± 2.0 ng/gm of tissue wet weight). The severity of OA cartilage lesions was similar in groups 1 and 2. In contrast, group 3 dogs had a marked reduction in macroscopic lesion severity on the tibial plateaus (P < 0.01 for grade; P < 0.04 for size) and femoral condyles. Moreover, the histologic lesion severity was decreased on both plateaus (P < 0.06) and condyles.

Conclusion. This study showed that a local increase in IL‐1Ra production in OA knee joints by intraarticular injection of transduced synovial cells can reduce the progression of experimentally induced lesions.
Interleukin‐17 up‐regulation of nitric oxide production in human osteoarthritis cartilage
Authors: Attur Mukundan G. , Patel Rajesh N. , Abramson Steven B. , Amin Ashok R.
Published: February 2007

Objective. To examine the effect of human interleukin‐17 (IL‐17) on nitric oxide (NO) production in human osteoarthritis (OA) cartilage under ex vivo conditions.

Methods. OA cartilage from patients undergoing knee replacement surgery was used in explant assays to assess the effect of IL‐17. NO production was measured by estimating the stable NO metabolite, nitrite, in conditioned medium.

Results. IL‐17 augmented the spontaneous production of nitric oxide. This augmentation was sensitive to cycloheximide and pyrrolidine dithiocarbamate, but not to dexamethasone or soluble IL‐1 receptor.

Conclusion. IL‐17 augments nitric oxide production in OA cartilage via nuclear factor KB activation, but independently of IL‐1β signaling.
Rheumatic diseases in an MRL strain of mice with a deficit in the functional Fas ligand
Authors: Ito Mituko R. , Terasaki Shizuko , Itoh Junpei , Katoh Hideki , Yonehara Shin , Nose Masato
Published: February 2007

Objective. To characterize Fas antigen expression on the cell surface, and to determine the effect of this expression in rheumatic diseases using a newly established gld‐congenic MRL strain of mice (MRL/gld), which is defective in its functional Fas ligand (Fas‐L).

Methods. Flow cytometric analyses of lymphoid cells and macrophages were performed using anti‐Fas and other cell surface markers. Histopathologic manifestations were examined using immunochemistry and light and electron microscopy. Serum levels of IgG and anti‐DNA antibodies were measured by single radial immunodiffusion and enzyme‐linked immunosorbent assay, respectively.

Results. MRL/gld mice developed systemic lymph‐adenopathy with an accumulation of Thy1.2+, B220+ and CD4‐, CD8‐ T cells, which both express the Fas antigen. Splenic B cells positive for surface IgM and/or surface IgD, and resident peritoneal macrophages exhibited up‐regulated expression of the Fas antigen, at much higher levels than those observed in MRL/MpJ‐+/+ (MRL/+) mice. Forms of rheumatic disease were observed in these mice, although not in C3H/HeJ‐gld/gld mice. These forms included diffuse glomerulonephritis, granulomatous arteritis, and arthritis, and were associated with the infiltration of mononuclear cells expressing the Fas antigen. Serum levels of IgG and anti‐DNA antibodies were significantly increased in MRL/gld mice compared with MRL/+ mice.

Conclusion. Rheumatic disease was generated by the gld gene in mice with an MRL background, as it is by the lpr gene, which is a Fas deletion mutant, associated with autoimmune traits. Rheumatic disease in this MRL strain was initiated by an incapacity for Fas/Fas‐L‐induced apoptosis, resulting in the development of autoimmunity and allowing for a persistent immune response in the affected lesions.
Impaired nonrestricted cytolytic activity in systemic lupus erythematosus
Authors: Stohl William , Elliott Julie E. , Li Lily , Podack Eckhard R. , Lynch David H. , Jacob Chaim O.
Published: February 2007

Objective. To determine the cytolytic effector pathway involved in impaired generation of nonrestricted cytolytic activity in systemic lupus erythematosus (SLE).

Methods. Peripheral blood mononuclear cells (PBMC) from normal subjects and SLE patients were stimulated in vitro with anti‐CD3 monoclonal antibody (MAb) and interleukin‐2 to promote the generation of nonrestricted cytolytic activity. On day 13 of culture, the PBMC were assayed for cytolytic activity against Fas‐Daudi cells and Fas+ Jurkat cells. The effects on cytotoxicity of calcium chelation, antagonist anti‐Fas MAb, tumor necrosis factor (TNF) α and β, and ATP were measured. Intracellular perforin expression was determined by intracellular staining, and perforin messenger RNA levels were determined by quantitative competitive reverse transcription‐polymerase chain reaction.

Results. We demonstrated the existence of a cytolytic pathway that is independent of Fas, TNFα, TNFβ, and ATP, but is dependent upon extracellular calcium. Despite its calcium dependence, this pathway is associated with low‐to‐undetectable levels of perforin.

Conclusion. Impaired generation of nonrestricted cytolytic activity in SLE is likely due to decreased activity of this Fas‐, TNFα‐, TNFβ‐, ATP‐independent pathway associated with very low levels of perforin.
Molecular analysis of major histocompatibility complex allelic associations with systemic lupus erythematosus in Taiwan
Authors: Lu Ling‐Ying , Ding Wei‐Zi , Fici Dolores , Deulofeut Richard , Cheng He‐Hsiung , Cheu Ching‐Cheng , Sung Ping‐Kuang , Schur Peter H. , Fraser Patricia A.
Published: February 2007

Objective. To investigate the association of HLA class II alleles/haplotypes, type I C2 deficiency gene, and tumor necrosis factor α gene promoter allele (TNF2) with systemic lupus erythematosus (SLE) in the Chinese population in Taiwan.

Methods. The HLA‐DRB1 and DQB1 alleles were studied in 105 SLE patients and 115 controls by the polymerase chain reaction (PCR)/sequence‐specific oligonucleotide probe method, the subtyping of DRB1*15/16 and DRB5 by PCR with sequence‐specific primers, type I C2 deficiency gene by PCR, and TNF2 by PCR‐Nco I restriction fragment length polymorphism.

Results. The frequencies of the HLA class II alleles DRB1*02, DRB1*1502, DRB5*0102, DQB1*0501, and DQB1*0602 and DR2‐associated haplotypes DRB1* 1501,DRB5*0101,DQB1*0602 and DRB1*1502,DRB5* 0102,DQB1*0501 were higher among SLE patients than among controls; however, only DQB1*0501 was statistically significantly associated with SLE. No specific allele/haplotype was significantly associated with lupus nephritis. No subject had type I C2 deficiency. SLE patients had a marginally higher percentage of TNF2, which was in linkage disequilibrium with DR3. Since DR3 was not associated with SLE in this Taiwanese Chinese population, TNF2 might play a role in the immunopathogenesis of SLE.

Conclusion. Although no HLA‐DRB1 allele was found to be significantly associated with SLE, the associations with DQB1*0501 and TNF2 suggest that DQB1 and tumor necrosis factor α may be important genetic factors in SLE susceptibility in the Chinese population in Taiwan.
The palpable tendon friction rub
Authors: Steen Virginia D. , Medsger Thomas A.
Published: February 2007

Objective. To evaluate the clinical and prognostic significance of palpable tendon friction rubs in patients with systemic sclerosis (SSc).

Methods. SSc patients evaluated prospectively at the University of Pittsburgh were examined serially for the presence of tendon friction rubs on physical examination. Demographic, clinical, and laboratory features of disease were obtained by patient examination, annual patient questionnaire, and medical record review. Patients were classified as having limited or diffuse scleroderma according to standard definitions. The prognostic significance of the presence of tendon friction rubs was determined using this comprehensive database.

Results. The SSc patients (n = 1,305) were first evaluated during 1972 through 1991 and were followed up for a mean of 6.3 years. Tendon friction rubs were detected most frequently in patients who had or who developed diffuse cutaneous involvement. There were strong correlations between the presence of tendon friction rubs and symptoms and signs typical of diffuse scleroderma, including more severe skin thickening, more frequent heart and kidney involvement, and decreased survival. In multiple regression analyses, the presence of 1 or more tendon friction rubs was one of the best predictors of both evolution to diffuse scleroderma and reduced survival.

Conclusion. The palpable tendon friction rub is an easily detected, inexpensively obtained physical examination finding which is highly associated with diffuse cutaneous scleroderma and decreased survival. This observation should lead to the early diagnosis of diffuse scleroderma and should identify patients at high risk for serious visceral involvement who are thus candidates for potential disease‐modifying therapy.
Interleukin‐1β‐stimulated invasion of articular cartilage by rheumatoid synovial fibroblasts is inhibited by antibodies to specific integrin receptors and by collagenase inhibitors
Authors: Wang Allan Z. , Wang Jane C. , Fisher Gregory W. , Diamond Herbert S.
Published: February 2007

Objective. To study the role of integrin receptors in the invasion of cartilage by rheumatoid synovial fibroblasts (RSF).

Methods. RSF were cocultured with cartilage slices alone or in the presence of various potential activators or inhibitors. The penetration of the cartilage surface by RSF was determined by live‐cell imaging of fluorescent‐labeled cells.

Results. Interleukin‐1β (IL‐1β) and IL‐8 stimulated the RSF invasion of cartilage. Invasion was specific for RSF and required a concentration gradient of IL‐1β. The IL‐1β‐activated invasion of cartilage was inhibited by anti‐IL‐1 antibodies, IL‐1 receptor antagonist, and collagenase inhibitors. RSF invasion was also inhibited by antibodies to α4, α5, αV, and β1 integrins.

Conclusion. In this study, an IL‐1β concentration gradient was required for RSF invasion into cartilage, raising the possibility that in vivo invasion may be induced by IL‐1β released by chondrocytes. The IL‐1β activation of RSF assayed in vitro may contribute to the RSF invasion of cartilage in vivo. Cartilage invasion requires the availability of β1 and α4, α5, and αV integrins and the presence of collagenase activity.
Mechanisms of drug‐induced lupus. IV. Comparison of procainamide and hydralazine with analogs in vitro and in vivo
Authors: Yung Raymond , Chang Sandra , Hemati Nahid , Johnson Kent , Richardson Bruce
Published: February 2007

Objective. T cells treated with DNA methylation inhibitors overexpress lymphocyte function‐associated antigen 1 (LFA‐1), which results in autoreactivity, and the autoreactive cells cause a lupus‐like disease in vivo, suggesting a mechanism by which some agents may cause drug‐induced lupus. This study compared the effects of procainamide (Pca) and hydralazine (Hyd) with those of structural analogs, to determine if the degree of LFA‐1 overexpression and T cell autoreactivity correlated with the ability of the agents to induce autoimmunity.

Methods. Cloned murine T helper 2 cells were treated with Pca, N‐acetylprocainamide, Hyd, Phthalazine, or hydroxyurea (HU). The treated cells were then compared for LFA‐1 overexpression, autoreactivity, and the ability to induce autoimmunity in vivo.

Results. Pca and Hyd were more potent than their analogs or HU in all 3 assays.

Conclusion. The results support a relationship between LFA‐1 overexpression, T cell autoreactivity, and autoimmunity, and suggest a mechanism by which Pca and Hyd, but not the analogs, may cause drug‐induced lupus.
Incidence of hydroxychloroquine retinopathy in 1,207 patients in a large multicenter outpatient practice
Authors: Levy G. D. , Munz S. J. , Paschal J. , Cohen H. B. , Pince K. J. , Peterson T.
Published: February 2007

Objective. To define the true risk of hydroxychloroquine (HCQ) retinal toxicity by studying the largest single group yet evaluated.

Methods. Retrospective chart review of all patients in the Kaiser Permanente Medical Care Program, Southern California Region, who had HCQ prescriptions filled from 1991 through 1993 (1,556 patients in 11 medical centers). Of 1,207 charts of patients who took HCQ and had documented ophthalmologic examinations, initial screening identified 21 charts (1.7%) that indicated possible HCQ toxicity.

Results. We identified 1 patient with definite toxicity (1 of 1,207; 0.08%) and 5 other patients with indeterminate but probable toxicity (5 of 1,207; 0.4%). The incidence of definite HCQ retinal toxicity in patients treated with HCQ at <6.5 mg/kg/day was 0.

Conclusion. In HCQ‐treated patients whose renal function is normal, routine ophthalmic screening is not indicated if the daily dosage is <6.5 mg/kg. In patients whose daily dosage is >6.5 mg/kg or who have taken HCQ continuously for > 10 years, annual screening may be appropriate.
Deposition of antibodies to the collagen‐like region of C1Q in renal glomeruli of patients with proliferative lupus glomerulonephritis
Authors: Mannik Mart , Wener Mark H.
Published: February 2007

Objective. To determine if antibodies to the collagen‐like region of C1q (C1q‐CLR) are present in the glomerular immune deposits of patients with systemic lupus erythematosus (SLE).

Methods. Kidney tissues were obtained at autopsy, glomeruli were isolated, and glomerular basement membrane fragments were prepared. Antibodies were extracted with low pH or with DNase.

Results. The concentrations of antibodies to C1q‐CLR recovered from the glomeruli were ⩾50‐fold higher per unit of IgG than that found in the serum or in the serum and interstitial fluid entrained in glomeruli. Antibodies to C1q‐CLR were recovered from glomeruli of 4 of 5 patients with proliferative glomerulonephritis at autopsy.

Conclusion. This is the first demonstration that antibodies to C1q‐CLR are deposited and concentrated in the renal glomeruli of patients with SLE. These antibodies, thus, have the potential of contributing to the pathogenesis of lupus glomerulonephritis.
Health status and disease severity in fibromyalgia. Results of a six‐center longitudinal study
Authors: Wolfe Frederick , Anderson Janice , Harkness Deborah , Bennett Robert M. , Caro Xavier J. , Goldenberg Don L. , Russell I. Jon , Yunus Muhammad B.
Published: February 2007

Objective. To determine the intermediate and long‐term outcomes of fibromyalgia in patients seen in rheumatology centers in which there is special interest in the syndrome.

Methods. We conducted a longitudinal outcome study by mailed comprehensive Health Assessment Questionnaire administered every 6 months to 538 patients, from 6 rheumatology centers, whose median duration of disease at first assessment was 7.8 years. The final assessment took place after 7 years. In addition, there was study followup on 85 patients who had attended the Wichita center for > 10 years.

Results. Although functional disability worsened slightly and health satisfaction improved slightly, measures of pain, global severity, fatigue, sleep disturbance, anxiety, depression, and health status were markedly abnormal at study initiation and were essentially unchanged over the study period. Correlations between first and last assessment values were as high as r = 0.82. For some variables, abnormalities were 3 times greater at one center compared with another.

Conclusion. Patients with established fibromyalgia, seen in rheumatology centers in which there a special interest in the disease and followed up for as long as 7 years, have markedly abnormal scores for pain, functional disability, fatigue, sleep disturbance, and psychological status, and these values do not change substantially over time. Half the patients are dissatisfied with their health, and 59% rate their health as fair or poor. There are marked differences in disease severity among the various centers, but <14% of the variance in outcomes can be explained by demographic or center factors. Values at the first assessment are predictive of final values.
Range of antinuclear antibodies in “healthy” individuals
Authors: Tan E. M. , Feltkamp T. E. W. , Smolen J. S. , Butcher B. , Dawkins R. , Fritzler M. J. , Gordon T. , Hardin J. A. , Kalden J. R. , Lahita R. G. , Maini R. N. , McDougal J. S. , Rothfield N. F. , Smeenk R. J. , Takasaki Y. , Wiik A. , Wilson M. R. , Koziol J. A.
Published: February 2007

Objective. To determine the range of antinuclear antibodies (ANA) in “healthy” individuals compared with that in patients with systemic lupus erythematosus (SLE), systemic sclerosis (SSc; scleroderma), Sjögren's syndrome (SS), rheumatoid arthritis (RA), or soft tissue rheumatism (STR).

Methods. Fifteen international laboratories experienced in performing tests for ANA by indirect immunofluorescence participated in analyzing coded sera from healthy individuals and from patients in the 5 different disease groups described above. Except for the stipulation that HEp‐2 cells should be used as substrate, each laboratory used its own in‐house methodology so that the data might be expected to reflect the output of a cross‐section of worldwide ANA reference laboratories. The sera were analyzed at 4 dilutions: 1:40, 1:80, 1:160, and 1:320.

Results. In healthy individuals, the frequency of ANA did not differ significantly across the 4 age subgroups spanning 20–60 years of age. This putatively normal population was ANA positive in 31.7% of individuals at 1:40 serum dilution, 13.3% at 1:80, 5.0% at 1:160, and 3.3% at 1:320. In comparison with the findings among the disease groups, a low cutoff point at 1:40 serum dilution (high sensitivity, low specificity) could have diagnostic value, since it would classify virtually all patients with SLE, SSc, or SS as ANA positive. Conversely, a high positive cutoff at 1:160 serum dilution (high specificity, low sensitivity) would be useful to confirm the presence of disease in only a portion of cases, but would be likely to exclude 95% of normal individuals.

Conclusion. It is recommended that laboratories performing immunofluorescent ANA tests should report results at both the 1:40 and 1:160 dilutions, and should supply information on the percentage of normal individuals who are positive at these dilutions. A low‐titer ANA is not necessarily insignificant and might depend on at least 4 specific factors. ANA assays can be a useful discriminant in recognizing certain disease conditions, but can create misunderstanding when the limitations are not fully appreciated.
Mono‐iodoacetate‐induced experimental osteoarthritis. A dose‐response study of loss of mobility, morphology, and biochemistry
Authors: Guingamp Corinne , Gegout‐Pottie Pascale , Philippe Lionel , Terlain Bernard , Netter Patrick , Gillet Pierre
Published: February 2007

Objective. To characterize the doseresponsiveness of morphologic and biochemical chondral changes relative to mobility in mono‐iodoacetate (MIA)‐induced osteoarthritis (OA) in rats.

Methods. Rat mobility was assessed by biotelemetry. Articular lesions were characterized by macroscopic and histologic examinations. Cartilage proteoglycan metabolism was evaluated by the 1,9‐dimethylmethylene blue dye binding assay and by radiosulfate incorporation in patellar cartilage.

Results. Spontaneous locomotor activity was rapidly, transiently, and dose‐dependently decreased after MIA injection into rat knees (primary response). Thereafter, only high doses (0.3 mg and 3.0 mg) led to a secondary progressive long‐term loss of spontaneous mobility on day 15, when subchondral bone was exposed. These 2 doses resulted in significant changes in cartilage proteoglycan concentration at day 15 and a strong inhibition of anabolism in the peripheral patellae by day 2, contrasting with the effects of lower doses (0.01, 0.03, and 0.1 mg).

Conclusion. When a sufficient dose of MIA is used, this model can easily and quickly reproduce OA‐like lesions and functional impairment in rats, similar to that observed in human disease. These parameters, as well as proteoglycan metabolism, could serve as indicators for studying chondroprotective drugs, or for evaluating the ability of imaging techniques to detect and evaluate chondral lesions.
Molecular cloning of a novel 97‐kd Golgi complex autoantigen associated with Sjögren's syndrome
Authors: Griffith Kevin J. , Chan Edward K. L. , Lung Chien‐Cheng , Hamel John C. , Guo Xiaoying , Miyachi Kiyomitsu , Fritzler Marvin J.
Published: February 2007

Objective. To identify a Golgi complex autoantigen bound by Sjögren's syndrome (SS) autoantibodies.

Methods. Serum from a patient with secondary SS and anti‐Golgi antibodies was used as a probe to isolate a complementary DNA (cDNA) insert from a HeLa cDNA library.

Results. A 3.7‐kb cDNA encoding a 56‐kd recombinant protein was immunoprecipitated by the human anti‐Golgi serum and immune rabbit serum. Western blot analysis showed that the immune rabbit sera recognized a protein of 97 kd (golgin‐97), suggesting that the isolated clone contained a partial cDNA. The 5′ upstream sequence was obtained by rapid amplification of the cDNA ends. The complete cDNA contained 4,860 basepairs, encoding a protein with a calculated Mr of 88 kd. Antibodies to golgin‐97 were found in 12 (20%) of 60 sera known to have anti‐Golgi autoantibodies, and the majority of these sera (8 of 12, or 75%) were from patients who had secondary SS.

Conclusion. Golgin‐97 is a unique Golgi complex antigen that appears to be a target of SS autoantibodies.