bluuebunny/crossref_metadata_2025 huggingface dataset:
Colchicine in breast milk of patients with familial mediterranean fever
Authors: Ben‐Chetrit Eldad , Scherrmann Jean‐Michel , Levy Micha
Published: February 2007

Objective. To clarify whether colchicine is excreted in breast milk, and to compare its concentrations in the serum and breast milk of lactating women who have familial Mediterranean fever (FMF).

Methods. Using a specific radioimmunoassay, we determined colchicine concentrations in the serum and breast milk of 4 patients at various time points, following oral administration of the drug. The study evaluated 4 patients with FMF who had been taking colchicine on a long‐term basis.

Results. Colchicine was found to be excreted in breast milk. Its levels ranged between 1.9 and 8.6 ng/ml, which were similar to those found in the serum (parallel concentration time curves). However, there appeared to be a considerable variation in colchicine milk concentration among the different patients, which might be related to individual breast milk composition and, possibly, to other nutritional or metabolic factors.

Conclusion. The extensive peripheral tissue binding and relatively low concentration of colchicine in breast milk suggests that the amount ingested by the infant is small. Furthermore, based on our clinical experience, nursing appears to be safe for lactating women with FMF who continue to take colchicine.
Pediatric rheumatology in adult rheumatology practices in Washington state
Authors: Sherry David D. , Wallace Carol A. , Kahn Stuart J.
Published: February 2007

Objective. To determine both the extent to which adult rheumatologists treat children and their level of comfort in doing so.

Methods. A questionnaire was sent to all 77 physicians in the state of Washington who were listed as adult rheumatologists in the American College of Rheumatology (ACR) directory.

Results. Sixty‐six questionnaires (86%) were returned; 50 were identified as being from privatepracticing adult rheumatologists and were the focus of this study. Sixty‐two percent of the respondents reported that they care for children; predictors included increased exposure to pediatric rheumatology during fellowship (P = 0.003), increased distance from Seattle (P = 0.001), and listing oneself in the ACR directory as treating children (P = 0.03). Most respondents reported feeling discomfort in treating children younger than 6 years of age, treating Kawasaki disease, and treating polyarteritis nodosa, but most reported feeling comfortable treating children with chronic arthritides. Impediments to referring to a pediatric rheumatologist included distance (median distance 35 miles), convenience for the family, personal preference, and experience in caring for children. Twenty‐nine percent reported difficulties referring to a pediatric rheumatologist outside of one's managed care plan. Adult rheumatologists expressed interest in continuing medical education dealing with pediatric rheumatology, preferably with a lecture format in their home communities.

Conclusion. A significant number of adult rheumatologists care for children. Pediatric rheumatologists should provide both educational and consultative support for these adult rheumatologist colleagues.
A matrix metalloproteinase–generated aggrecan neoepitope as a marker of skeletal maturation and aging in cartilage
Authors: Olszewski Julie , McDonnell Joseph , Stevens Karla , Visco Denise , Moore Vernon
Published: February 2007

Objective. To quantify the matrix metalloproteinase–induced neoepitope F(M/V)DIPEN (Phe–[Met/Vall]‐Asp–Jle–Pro–Glu–Asn341) in guinea pig and rabbit cartilage during aging.

Methods. Cartilage was taken from the stifle joint, nasal septum, and xiphoid process in guinea pigs and rabbits at selected ages. The cartilage was then extracted and evaluated for F(M/V)DIPEN levels by radioimmunoassay.

Results. In the 3 tissues studied, there were major increases in F(M/V)DIPEN levels during skeletal maturation and aging in both the guinea pig and rabbit cartilage. Except for spontaneous osteoarthritis that develops in guinea pigs with aging, increases in the neoepitope were not correlated with arthritis pathology.

Conclusion. Increases in the level of F(M/V)‐DIPEN in cartilage occur as a result of skeletal maturation and aging. This physiologic accumulation of F(M/V)DIPEN in cartilage should be considered when using the neoepitope as a disease marker in arthritis.
Focal myositis presenting as pseudothrombophlebitis of the neck in a patient with mixed connective tissue disease
Authors: Rivest Charles , Miller Frederick W. , Love Lori A. , Turgeon Pierre‐Paul , Blier Claude , Senécal Jean‐Luc
Published: February 2007

This report describes a case of focal myositis in a patient with mixed connective tissue disease. The patient presented with diffuse neck swelling and pseudothrombophlebitis of the left internal jugular vein. Other clinical features included a high fever, elevated erythrocyte sedimentation rate, and prompt improvement after administration of high‐dose intravenous corticosteroid therapy. Criteria for polymyositis were absent, serum levels of creatine kinase remained normal, and there was no sign of recurrence during 3 years of followup. Results of immunoprecipitation for anti–Jo‐1 and other myositis‐specific autoantibodies remained negative in serial serum samples obtained before, during, and after the episode.
The CD69 activation pathway in rheumatoid arthritis synovial fluid t cells
Authors: Hernández‐García César , Fernández‐Gutiérrez Benjamín , Morado Inmaculada C. , Bañares Antonio A. , Jover Juan A.
Published: February 2007

Objective. To study the CD69 activation pathway in synovial fluid (SF) T lymphocytes from patients with rheumatoid arthritis (RA).

Methods. Peripheral blood mononuclear cells (PBMC) or SF mononuclear cells (SFMC) were used in proliferation assays with anti‐CD69, anti‐CD28, anti‐CD3, phorbol myristate acetate (PMA), and/or recombinant interleukin‐2 (IL‐2). CD69+, CD69—, and resting SF T cells were also proliferated. CD25 expression and production of IL‐2 after CD69 activation were assessed by flow cytometry and in a bioassay with the IL‐2‐dependent cell line CTLL‐2.

Results. RA SFMC did not proliferate either in the presence of anti‐CD69 monoclonal antibodies alone or with concomitant PMA activation, when compared with paired or control PBMC. Similar low proliferative responses via the CD3 or CD28 pathway with PMA were observed. This defective proliferation of RA SFMC after stimulation through the CD69 molecule was explained in part by a failure to express CD25 and to produce IL‐2. SF CD69‐ T cells and resting SF T cells had higher rates of proliferation through the alternative costimulatory pathway CD28 than did SF CD69+ T cells or freshly isolated SF T cells.

Conclusion. Freshly isolated SF T cells present a profound state of hyporesponsiveness through the CD69 and CD28 costimulatory pathways. This state appears to be dependent on the activation status of SF T cells, since CD69— and resting SF T cells showed recovery of the ability to proliferate through the CD28 activation pathway.
Heterogeneity of collagen synthesis in normal and systemic sclerosis skin fibroblasts. Increased proportion of high collagen‐producing cells in systemic sclerosis fibroblasts
Authors: Jelaska Ante , Arakawa Masami , Broketa Goran , Korn Joseph H.
Published: February 2007

Objective. The goal of this study was to quantitatively analyze the distribution of collagen synthesis in normal and systemic sclerosis (SSc) fibroblast populations in order to determine the extent of activation in SSc populations.

Methods. We used quantitative in situ hybridization to assess the population distribution of type I collagen synthesis. Fibroblast cultures were derived from both clinically involved and uninvolved skin regions of patients with SSc, and from healthy adults, and assessed for levels of α1(I) procollagen messenger RNA (mRNA).

Results. Dermal fibroblasts from both patients with SSc and normal adults were heterogeneous for distribution of α1(I) procollagen mRNA when assessed by in situ hybridization, with a wide range of grains per cell. In contrast, clones of neonatal fibroblasts showed a relatively homogeneous distribution of grain counts. Involved SSc skin fibroblasts had a larger proportion of cells in the high collagen‐producing mRNA subpopulation (mean ± SEM 28.4 ± 6.85%), compared with normal fibroblasts (1.75 ± 1.44%) and uninvolved fibroblasts (9.6 ± 6.73%). Conversely, within the low collagen‐producing mRNA subpopulation, involved fibroblasts had a smaller proportion of cells (mean ± SEM 14.0 ± 5.63%) than did uninvolved fibroblasts (37.8 ± 13.69%), while normal fibroblasts had a majority of the cells in this subpopulation (53.5 ± 8.68%).

Conclusion. These results suggest that only a specific subset of fibroblasts are activated in SSc, as evidenced by an increased proportion of cells with high levels of α1(I) procollagen mRNA. Differences between the SSc and normal fibroblast populations appeared to be quantitative rather than qualitative. This may be a result of either clonal selection or selective activation in the pathogenesis of SSc.
Transcriptional activation of the α1(1) procollagen gene in systemic sclerosis dermal fibroblasts. Role of intronic sequences
Authors: Hitraya Elena G. , Jiménez Sergio A.
Published: February 2007

Objective. To investigate the transcriptional regulation of the α(1) procollagen gene (COL1A1) in cultured dermal fibroblasts from patients with diffuse systemic sclerosis (SSc) of recent onset and to evaluate the role that intronic sequences may play in the upregulated expression of COL1A1 in SSc dermal fibroblasts.

Methods. Dermal fibroblasts from 6 patients with diffuse SSc of recent onset and from 3 healthy individuals were studied. The steady‐state levels of α1(I) procollagen messenger RNA were evaluated by Northern hybridization analysis, and the transcriptional regulation of COL1A1 was examined by transient transfection experiments with deletion constructs containing portions of COL1A1 promoter (with 5′ end points at ‐5.3 kb, ‐2.3 kb, and ‐804 bp and 3′ end point at +42 bp) ligated to the chloramphenicol acetyltransferase (CAT) reporter gene. To examine the role of intronic sequences, constructs containing, in addition to the COL1A1 promoter, a portion of the first intron (+380 bp to +1,440 bp) cloned in front of the CAT gene were transfected. The efficiency of transfections was normalized relative to the net amount of CAT plasmid actually transfected into recipient cells, determined by a modified Southern hybridization procedure.

Results. Maximal CAT activity was observed with constructs extending from ‐804 bp to +42 bp in both normal and SSc fibroblasts. However, the activity driven by this construct was 80–110% higher in SSc fibroblasts. The CAT activity driven by a construct with a 5′ end point at ‐5.3 kb was only 15–20% higher in SSc cells, and the CAT activity driven by a construct with a 5′ end point ‐5.3 kb was only 15–20% higher in SSc cells, and the CAT activity driven by a construct with a 5′ end point ‐2.3 kb was 35‐45% higher in SSc fibroblasts. The CAT activity driven by the ‐804‐bp promoter construct was increased up to 4‐fold in SSc fibroblasts in comparison with normal cells when the intronic segment spanning +380 bp to +1,440 bp was included in the transfected construct.

Conclusion. The results directly demonstrate the transcriptional activation of COL1A1 in dermal fibroblasts from SSc patients. The data also indicate that first‐intron sequences of COL1A1 are required for maximal transcriptional activity of the collagen gene and may play an important role in the up‐regulation of its expression in SSc fibroblasts.
Development of complete heart block in an adult patient with Sjögren's syndrome and anti‐Ro/SS‐A autoantibodies
Authors: Lee Lela A. , Pickrell Michael B. , Reichlin Morris
Published: February 2007

We describe the occurrence of complete heart block in a patient with Sjögren's syndrome. The patient's serum contained antibodies to both the 60‐kd and 52‐kd Ro proteins. This case indicates that although the adult atrioventricular node may be relatively resistant to the development of anti‐Ro–associated heart block, it can nevertheless be affected.
Increased responsiveness of rheumatoid factor–producing B cells in seronegative and seropositive rheumatoid arthritis
Authors: He Xiaowen , Zhong Wanyun , McCarthy Timothy G. , Weyand Cornelia M. , Goronzy Jörg J.
Published: February 2007

Objective. To compare the frequencies and responsiveness of rheumatoid factor (RF)–producing B cells in the peripheral blood of patients with seronegative and seropositive rheumatoid arthritis (RA).

Methods. Frequencies of IgM+, IgG+, and RF+ B cells were determined by limiting‐dilution analysis of purified peripheral blood B cells from 6 patients with seropositive RA, 8 patients with seronegative RA, and 7 normal controls. B cell help was provided by cloned T helper cells, which were stimulated by either anti‐CD3 or the bacterial superantigen staphylococcal enterotoxin D (SED). IgM and IgG antibodies and RF in culture supernatants were detected by enzyme‐linked immunosorbent assay.

Results. In the presence of anti‐CD3–stimulated T helper cells, 2–10% of B cells from normal individuals secreted IgM and IgG antibodies. The frequency of RF+ B cells was low and ranged from 1:182 to 1:885 (RF+: IgM+) B cells. In patients with seropositive RA, the numbers of Ig‐producing B cells were reduced by a factor of 2, while the fraction of RF+ B cell precursors was expanded by more than 50‐fold (7–20% of IgM+ B cells; P = 0.004). Patients with seronegative RA had higher frequencies of RF‐producing B cells (1.5–6% of IgM+ B cells) than normal individuals (P = 0.002), but not to the same extent as seropositive patients (P = 0.002). Stimulation of B cells using SED preferentially induced RF+ B cells in normal controls and in patients with seronegative and seropositive RA.

Conclusion. B cell precursors with the potential to secrete RF were detectable in high frequencies in normal individuals and in patients with seropositive and seronegative RA. In all donors, these B cells could be stimulated with the bacterial superantigen SED. In normal individuals, RF+ B cells remained nonresponsive to help provided by anti‐CD3–activated T cells, but were responsive in RA patients. Seronegative and seropositive RA form a continuous spectrum of disease, with a higher number of RF‐secreting B cells in the seropositive patients.
Chondroprotective effect of intraarticular injections of interleukin‐1 receptor antagonist in experimental osteoarthritis. Suppression of collagenase‐1 expression
Authors: Caron John P. , Fernandes Julio C. , Martel‐Pelletier Johanne , Tardif Ginette , Mineau François , Geng Changshan , Pelletier Jean‐Pierre
Published: February 2007

Objective. To investigate the in vivo effect of recombinant human interleukin‐1 receptor antagonist (rHuIL‐1Ra) on the development of lesions and the expression of metalloproteases in the canine experimental osteoarthritis (OA) model.

Methods. The right anterior cruciate ligament was sectioned percutaneously in 3 groups of dogs. The control group (n = 5) received an intraarticular injection of sterile physiologic saline (1 ml) twice weekly for 4 weeks starting on the day of surgery. The remaining 2 groups received intraarticular injections of either 2 mg (n = 6) or 4 mg (n = 5) rHuIL‐1Ra in 1 ml of physiologic saline according to the same schedule as the first group. All dogs were killed 4 weeks after surgery. The macroscopic appearance of femoral condyle osteophytes and the size and severity of cartilage lesions on femoral condyles and tibial plateaus were evaluated, as were the histologic features of cartilage and synovial membrane. Levels of collagenase‐1 and stromelysin‐1 messenger RNA expression in cartilage and synovium were determined by Northern blotting.

Results. Recombinant human IL‐1Ra exerted a dose‐dependent protective effect on the development of osteophytes and cartilage lesions in vivo. Treatment with rHuIL‐1Ra reduced the incidence (saline‐treated group 70%, 2 mg rHuIL‐1Ra–treated group 42%, 4 mg rHuIL‐1Ra–treated group 20%) and size (saline‐treated group 2.3 ± 0.7 mm [mean ± SEM], 2 mg rHuIL‐1Ra–treated group 0.7 ± 0.3 mm, 4 mg rHuIL‐1Ra–treated group 0.5 ± 0.3 mm) of femoral condyle osteophytes. In addition, a dose‐dependent decrease in the size (saline‐treated group 24.40 ± 8.17 mm2, 2 mg rHuIL‐1Ra–treated group 20.90 ± 8.01 mm2, 4 mg rHuIL‐1Ra–treated group 7.70 ± 5.16 mm2) and the grade (0–4 scale; saline‐treated group 1.20 ± 0.29, 2 mg rHuIL‐1Ra–treated group 1.00 ± 0.26, 4 mg rHuIL‐1Ra–treated group 0.30 ± 0.21) of the tibial plateau cartilage lesions was found, with a significant difference (P < 0.04) reached only with 4 mg rHuIL‐1Ra. Similarly, the histologic lesions in dogs treated with 4 mg rHuIL‐1Ra (Mankin scale; mean ± SEM 2.95 ± 0.53) were significantly less severe (P < 0.002) compared with those in the saline‐treated group (4.95 ± 0.54). Importantly, rHuIL‐1Ra treatment led to a significant reduction (P < 0.005) of collagenase‐1 expression in OA cartilage.

Conclusion. This study demonstrated that intraarticular injections of rHuIL‐1Ra can protect against the development of experimentally induced OA lesions. This effect could result, at least in part, from a reduction of collagenase‐1 expression. However, other catabolic processes involved in the degradation of OA cartilage may also be affected.
Local disruption of the insulin‐like growth factor system in the arthritic joint
Authors: Fernihough J. K. , Billingham M. E. J. , Cwyfan‐Hughes S. , Holly J. M. P.
Published: February 2007

Objective. To identify differences in levels of insulin‐like growth factor (IGF) and IGF binding proteins (IG‐FBPs) between 30 patients with arthritis (14 with rheumatoid arthritis [RA], 16 with osteoarthritis [OA]) and 11 normal control subjects. IGF and IGFBP levels were correlated to the disease activity marker C‐reactive protein (CRP) to determine whether they were disease related. We also examined the degree of proteolytic modification of the IGFBPs.

Methods. Radioimmunoassays were used for measuring IGF and IGFBP‐3 levels; CRP was measured by enzyme‐linked immunosorbent assay. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis followed by Western blotting, chemiluminescence, and autoradiography were used for visualizing binding proteins.

Results. There was a significant increase in synovial fluid levels of both IGF‐1 and IGFBP‐3 in both RA and OA. This resulted in an elevated IGFBP‐3 to IGF molar ratio of 1.49 in the OA group and 1.47 in the RA group, compared with 0.86 in the normal control group (P = 0.0002 for both). A significantly lower degree of IGFBP‐3 proteolysis was also seen in the synovial fluids from the patients compared with the controls. There were significant correlations between the CRP level and levels of IGF‐1, IGF‐2, and IGFBP‐3 in the RA patients (r = 0.62–0.898, P = 0.04–0.0007).

Conclusion. There was significant local disruption of the IGF system in patients with arthritis. This may result in a lower amount of IGF that is able to bind to IGF receptors in the arthritic joint. Levels of IGF‐1, IGF‐2, and IGFBP‐3 all correlated with the CRP level in patients with RA, which indicates the possibility that the IGF system is involved in the disease process.
Hepatocyte growth factor. A cytokine mediating endothelial migration in inflammatory arthritis
Authors: Koch Alisa E. , Halloran Margaret M. , Hosaka Shigeru , Shah Manisha R. , Haskell Catherine J. , Baker Steven K. , Panos Ralph J. , Haines G. Kenneth , Bennett Gregory L. , Pope Richard M. , Ferrara Napoleone
Published: February 2007

Objective. Angiogenesis is an integral component of the vasculoproliferative phase of rheumatoid arthritis (RA). Recently, a heparin‐binding cytokine termed hepatocyte growth factor (HGF), or scatter factor (due to its ability to disperse cohesive epithelial colonies), was described. We conducted this study to investigate the hypothesis that this cytokine was present in the milieu of the inflamed joint, and that it contributed to the chemotaxis of endothelial cells in the synovial tissue.

Methods. We examined synovial fluid, synovial tissue, and peripheral blood from 91 patients with RA and other arthritides. We used 83 total samples in an enzyme‐linked immunosorbent assay to quantitate the HGF in synovial fluids and peripheral blood. To determine whether the HGF was biologically active, an epithelial scatter factor assay was performed. Immunohistochemical analysis was used to determine localization in synovial tissues. To define a function for synovial HGF, we preincubated rheumatoid synovial fluids with neutralizing anti‐HGF and measured the ability of these synovial fluids to induce endothelial chemotaxis.

Results. Synovial fluid from patients with RA contained a mean ± SEM HGF concentration of 2.0 ± 0.3 ng/ml, while synovial fluid from patients with other arthritides (including inflammatory arthritis) contained 2.4 ± 0.7 ng/ml HGF. Osteoarthritis (OA) patient samples contained the smallest quantities of synovial fluid HGF at 0.9 ± 0.1 ng/ml. RA synovial fluid contained significantly more HGF than did RA peripheral blood (1.1 ± 0.2 ng/ml) (P < 0.05). Rheumatoid synovial fluids induced more scattering of cells than did OA synovial fluids, suggesting a role for this cytokine in rheumatoid joint destruction. Interleukin‐1β induced expression of rheumatoid synovial tissue fibroblast antigenic HGF and scatter factor activity. Immunohistochemically, HGF, as well as the HGF receptor (the met gene product), localized to significantly more rheumatoid synovial tissue lining cells than normal lining cells (P < 0.05). Both HGF and its receptor immunolocalized to subsynovial macrophages as well. Levels of synovial tissue immunoreactive HGF correlated positively with the number of synovial tissue blood vessels. Anti‐HGF neutralized a mean of 24% of the chemotactic activity for endothelial cells found in 10 rheumatoid synovial fluid samples.

Conclusion. These results indicate that synovial HGF may contribute to the vasculoproliferative phase of inflammatory arthritides such as RA, by inducing HGF‐mediated synovial neovascularization. These findings point to a newly described role for HGF in the fibroproliferative phase of RA‐associated synovitis.
Expression of matrix metalloproteinase 9 (96‐kd gelatinase B) in human rheumatoid arthritis
Authors: Ahrens Diane , Koch Alisa E. , Pope Richard M. , Stein‐Picarella Monica , Niedbala Michael J.
Published: February 2007

Objective. To determine the expression of matrix metalloproteinase 9/gelatinase B (MMP‐9) in synovial fluid (SF), plasma, and synovial tissue from individuals with rheumatoid arthritis (RA), inflammatory arthritis (IA), and osteoarthritis (OA), using specific monoclonal antibody reagents.

Methods. Gelatinolytic activity in the SF and plasma of patients with RA, IA, and OA was assessed by gelatin zymography. A mouse monoclonal antiserum, 277.13, which selectively recognizes soluble latent forms of human MMP‐9, was used to quantitate MMP‐9 levels in patient synovial effusions, plasma, and synovial tissue with a capture sandwich enzyme‐linked immunosorbent assay (ELISA). Fifty‐one SF samples (31 RA, 9 OA, 11 IA) were analyzed. Immunolocalization of MMP‐9 in RA, OA, and normal synovium was investigated using MMP‐9–specific antisera.

Results. MMP‐9 antigen levels in synovial effusions were elevated 67‐fold in RA samples compared with OA samples. In addition, although MMP‐9 antigen levels in IA synovial effusions were 2.7‐fold less than the values in RA samples, they were elevated 34‐fold over the values in OA samples. These data indicate an association between increased MMP‐9 levels and inflammatory arthritis. A predominant 92‐kd gelatinolytic activity (specifically inhibited by EDTA) was evident in RA and IA samples, but no activity was observed in OA samples. Among 86 plasma samples (17 RA, 9 IA, 60 normal controls) analyzed for MMP‐9 antigen levels by immunocapture ELISA, MMP‐9 antigen levels were elevated 7‐fold in RA plasma compared with normal plasma. RA synovial tissue extracts demonstrated elevated levels of MMP‐9 antigen compared with OA synovial tissue. MMP‐9 immunolocalization studies demonstrated expression in infiltrating leukocytes (neutrophils and macrophages), endothelial cells, and synovial fibroblasts in RA synovium.

Conclusion. Latent MMP‐9 and/or MMP‐9–tissue inhibitor of metalloproteinases 1 (TIMP‐1) complexes are elevated in RA and IA SF compared with OA SF. In addition, MMP‐9 is increased in RA plasma versus normal control plasma. Synovial tissue levels of MMP‐9 antigen are also elevated in RA versus OA. The tissue distribution of MMP‐9 within RA synovium is localized to sites of inflammation comprising surface synovial lining cells, endothelium, and leukocytes. Taken together, these observations suggest that connective tissue turnover occurs as a result of excessive MMP activity over TIMP action in the invading pannus, periarticular tissue, or SF. Further studies such as those used in the present investigation will help elucidate the role of a number of different enzymes and inhibitors in the destructive arthropathies.
Selective antibody reactivity with peptides from human endogenous retroviruses and nonviral poly(amino acids) in patients with systemic lupus erythematosus
Authors: Bengtsson Anders , Blomberg Jonas , Nived Ola , Pipkorn Rüdiger , Toth Lajos , Sturfel Gunnar
Published: February 2007

Objective. To investigate antibody responses to a broad panel of peptides derived from human endogenous retroviruses (HERVs) among unselected patients with systemic lupus erythematosus (SLE).

Methods. In sera obtained from 69 patients with SLE and healthy blood donors, immunoassay was used to measure levels of antibody against synthetic peptides derived from HERVs, exogenous retroviruses, and nonviral poly(amino acids).

Results. Measurement by immunoassay revealed increased frequencies of antiretroviral antibodies against 2 peptides derived from the env gene of the type C—like class, which includes ERV‐9 and HERV‐H, and against 2 peptides from the gag region of human T lymphotropic virus type I—related endogenous sequence 1, in patients with SLE. Antibodies to 2 nonviral peptides, polyhistidine and polyproline, were also overrepresented in patient sera. In 1 patient, longitudinal data obtained over a period of 12 years indicated that the concentrations of certain antiretroviral antibodies varied according to disease activity.

Conclusion. Reactivity to certain type C HERV—derived antigens was found among patients with SLE. This reactivity could be explained by increased exposure to cross‐reactive epitopes from essentially complete type C HERVs.
Development of the anti‐Ro autoantibody response in a patient with systemic lupus erythematosus
Authors: Scofield R. Hal , Zhang Fenchung , Kurien Biji T. , Anderson Camille J. , Reichlin Morris , Harley John B. , Stafford Haraldine A.
Published: February 2007

Objective. To characterize the initial events in anti‐Ro production by a patient with systemic lupus erythematosus, in whom this autoantibody is developing.

Methods. The immune response to the Ro ribonucleoprotein and other autoantigens were studied by enzyme‐linked immunosorbent assay for IgG and IgM, by isoelectric focusing, and by inhibition studies to determine apparent avidity.

Results. The patient's sera showed an oligoclonal response to Ro that increased in complexity and affinity with time. IgM anti‐Ro appeared shortly before IgG anti‐Ro, and disappeared as IgG anti‐Ro increased in titer and affinity. IgG antiribosomal P autoantibodies also appeared during the patient's course, but in contrast to anti‐Ro, were not preceded by IgM antiribosomal P.

Conclusion. These data are consistent with the Ro autoantigen being presented and processed in a manner similar to heterologous antigen, and with differences in the mechanisms that lead to the production of IgG anti‐Ro autoantibodies as opposed to antiribosomal P autoantibodies.
Rheumatoid arthritis lung disease. Determinants of radiographic and physiologic abnormalities
Authors: Saag Kenneth G. , Kolluri Sheela , Koehnke Rachelle K. , Georgou Thomas A. , Rachow John W. , Hunninghake Gary W. , Schwartz David A.
Published: February 2007

Objective. To determine the prevalence and important clinical predictors of radiographic and physiologic abnormalities indicative of rheumatoid arthritis interstitial lung disease (RA‐ILD).

Methods. An unselected cohort of patients with a confirmed diagnosis of RA and known lung disease were identified (n = 336) and evaluated for RA disease activity and severity. Outcomes included abnormalities determined by the pulmonary function tests of forced vital capacity (FVC) and diffusion capacity for carbon monoxide (DLco), and/or chest radiographic findings of interstitial infiltrates. We used multivariable statistical modeling to determine the independent significance of cigarette smoking and other RA‐specific factors on the pulmonary abnormalities of interest.

Results. At least 1 of the 3 abnormal findings was identified by pulmonary tests in 32.4% of all patients. These abnormal findings included an FVC <80% of predicted in 42 patients, a DLco <80% of predicted in 64 patients, and evidence of radiographic interstitial infiltrates in 40 patients. After statistical adjustment for confounding factors, pack‐years of cigarette smoking remained a significant predictor of low DLco (β = −0.07, 95% confidence interval [95% CI] −0.09, −0.04), low FVC (β = −0.003, 95% CI −0.006, −0.0004), and interstitial abnormalities on chest radiograph (odds ratio for ⩾25 pack‐years = 3.76, 95% CI 1.59, 8.88). The Health Assessment Questionnaire (HAQ) Disability Index (DI) was also an important risk factor for the decline in both the DLco (β = −1.15, 95% CI −2.00, −0.30) and FVC (β = −0.23, 95% CI −0.32, −0.13).

Conclusion. Although RA disease activity/severity (particularly as defined by the HAQ DI) was important, smoking was the most consistent independent predictor of radiographic and physiologic abnormalities suggestive of ILD in RA.
Antibodies to type II collagen in early rheumatoid arthritis. Correlation with disease progression
Authors: Cook Andrew D. , Rowley Merrill J. , Mackay Ian R. , Gough Andrew , Emery Paul
Published: February 2007

Objective. To establish that frequencies and levels of IgG antibodies to type II collagen are higher in early rheumatoid arthritis (RA), and to correlate these results with disease activity.

Methods. Forty‐four patients were characterized as having early RA. Patient sera obtained at initial presentation and at 12 months were tested by enzyme‐linked immunosorbent assay for IgG antibodies to native and denatured type II collagen.

Results. IgG antibodies to native and denatured type II collagen were detected at initial presentation in 27% and 82% of patients, respectively, and after 12 months in 14% and 50%, respectively. The presence of antibodies to native collagen was associated with activity of RA and severity of symptoms, and loss of antibodies at 12 months was associated with initially erosive RA and the DRB1 disease susceptibility motif.

Conclusion. Levels of serum IgG antibodies to collagen in RA decrease over time and, therefore, are not attributable simply to cartilage destruction. The presence of early positivity for these antibodies, together with the RA susceptibility motif, appears to be predictive of rapidly progressive RA.
Comparison of synovial tissue and synovial fluid as the source of nucleic acids for detection of Chlamydia trachomatis by polymerase chain reaction
Authors: Branigan Patrick J. , Gérard Hervé C. , Hudson Alan P. , Schumacher H. Ralph
Published: February 2007

Objective. Difficulties in detecting Chlamydia trachomatis in human joints by polymerase chain reaction (PCR) may be related to whether synovial tissue or synovial fluid (SF) is used as the source of DNA in PCR amplification. In this study, a new PCR assay was developed and used to compare chlamydial DNA in paired samples of SF and synovial tissue from patients with arthritis.

Methods. The PCR assay targeted the ribosomal RNA operons, which are present in 2 copies on the C trachomatis chromosome. DNA from several relevant bacteria and chlamydial serovars was used for testing this screening system. The detection of chlamydial DNA in nucleic acid preparations from matched samples of SF and synovial tissue was compared by PCR assay. Samples were obtained from 55 patients, including patients with reactive arthritis, Reiter's syndrome, and other arthropathies.

Results. Testing of the PCR screening system confirmed it to be highly specific and sensitive. Use of this assay to screen DNA from SF and synovial tissue samples showed that 29 (53%) of 55 synovial tissue preparations were positive for chlamydial DNA, but only 16 (29%) of the matched SF samples from these 29 patients were similarly positive. Five (9%) of 55 SF samples, but not their tissue counterparts, were positive for chlamydial DNA by PCR.

Conclusion. Detection of chlamydial DNA in the joints of patients by PCR gives positive results more often when synovial tissue rather than SF is the source of target nucleic acids. Although synovial tissue is the source of choice for the most reliable determination of chlamydia in the joint, both synovial tissue and SF should be assayed if possible.
Effects of pulse methylprednisolone on cell adhesion molecules in the synovial membrane in rheumatoid arthritis. Reduced E‐selectin and intercellular adhesion molecule 1 expression
Authors: Youssef Peter P. , Triantafillou Sophie , Parker Angela , Coleman Mark , Roberts‐Thomson Peter J. , Ahern Michael J. , Smith Malcolm D.
Published: February 2007

Objective. To investigate the effects of a 1,000‐mg intravenous pulse of methylprednisolone succinate (MP) on cell adhesion molecule expression on the synovial vascular endothelium in patients with rheumatoid arthritis (RA).

Methods. Sequential arthroscopic biopsy samples were taken before and 24 hours after MP administration (10 patients) and at the time of RA flare (2 patients) and after retreatment with MP (1 patient). Immunoperoxidase staining for E‐selectin (CD62E), P‐selectin (CD62P), intercellular adhesion molecule 1 (ICAM‐1; CD54) and platelet—endothelial cell adhesion molecule (PECAM; CD31) was performed, and the staining was quantified by color video image analysis.

Results. MP caused a rapid (within 24 hours) and substantial decrease in the expression of E‐selectin on the synovial vascular endothelium, with a smaller reduction in ICAM‐1 expression on synovial vascular endothelium and the synovial lining. There were no similar effects on synovial membrane P‐selectin or PECAM expression.

Conclusion. A potential mechanism by which MP impairs neutrophil trafficking into inflamed RA joints might be by reducing E‐selectin, and possibly, ICAM‐1, expression in the synovial membrane.
Comparison of sulfasalazine and placebo in the treatment of reactive arthritis (Reiter's syndrome). A department of veterans affairs cooperative study
Authors: Clegg Daniel O. , Reda Domenic J. , Weisman Michael H. , Cush John J. , Vasey Frank B. , Schumacher H. Ralph , Budiman‐Mak Elly , Balestra Dominic J. , Blackburn Warren D. , Cannon Grant W. , Inman Robert D. , Alepa F. Paul , Mejias Edwin , Cohen Miriam R. , Makkena Rama , Mahowald Maren L. , Higashida Jean , Silverman Stuart L. , Parhami Nourollah , Buxbaum Joel , Haakenson Clair M. , Ward Richard H. , Manaster B. J. , Anderson Robert J. , Ward John R. , Henderson William G.
Published: February 2007

Objective. To determine whether sulfasalazine (SSZ) at a dosage of 2,000 mg/day is effective in the treatment of reactive arthritis (ReA) that has been unresponsive to nonsteroidal antiinflammatory drug (NSAID) therapy.

Methods. One hundred thirty‐four patients with ReA who had failed to respond to NSAIDs were recruited from 19 clinics, randomized (double‐blind) to receive either SSZ or placebo, and followed up for 36 weeks. The definition of treatment response was based on joint pain/tenderness and swelling scores and physician and patient global assessments.

Results. Longitudinal analysis revealed improvement in the patients taking SSZ compared with those taking placebo, which appeared at 4 weeks and continued through the trial (P = 0.02). At the end of treatment, response rates were 62.3% for SSZ treatment compared with 47.7% for placebo treatment. The Westergren erythrocyte sedimentation rate declined more with SSZ treatment than with placebo (P < 0.0001). Adverse reactions were fewer than expected and were mainly due to nonspecific gastrointestinal complaints.

Conclusion. SSZ at a dosage of 2,000 mg/day is well tolerated and effective in patients with chronically active ReA.
Preliminary experience with low‐dose methotrexate in fibroblastic rheumatism
Authors: Vittecoq Olivier , Mejjad Othmane , Silva Fernando Da , Joly Pascal , Thomine Elizabeth , Lauret Philippe , Thomine Jean Michel , Loët Xavier Le
Published: February 2007

Fibroblastic rheumatism (FR) is a rare disease characterized by joint manifestations without joint destruction, and associated with cutaneous nodules and sclerodactyly. The diagnosis is usually based on histologic examination of a skin nodule or synovium specimen. In the 11 previously reported cases, corticosteroid treatment, which has been widely used, was shown to have an unpredictable effect on the evolution of FR. We report a case of FR that was treated with methotrexate for 5 years, which led to complete recovery. This drug may be considered an effective treatment for FR. Further cases are needed to confirm this preliminary experience.
Immunohistologic analysis of renal CD40 and CD40L expression in lupus nephritis and other glomerulonephritides
Authors: Yellin Michael J. , D'Agati Vivette , Parkinson Glenn , Han Angelina Soh‐Yung , Szema Anthony , Baum David , Estes Dorothy , Szabolcs Matthias , Chess Leonard
Published: February 2007

Objective. To investigate potential mechanisms by which CD40L‐mediated signals may be involved in the pathogenesis of lupus glomerulonephritis (GN).

Methods. Renal in situ CD40L and CD40 expression was examined in patient biopsy specimens. Immuno‐histochemical studies were performed on frozen sections utilizing anti‐CD40L monoclonal antibody (MAb), anti‐CD40 MAb, or control MAb. As controls, we analyzed normal kidney specimens and specimens obtained from patients with IgA nephropathy, focal segmental glomerulosclerosis, minimal change disease, idiopathic membranous GN, and antineutrophil cytoplasmic antibody‐positive pauci‐immune GN. Staining distribution was noted and staining intensity scored on a semiquantitative scale of 0 (no staining) to 3+ (intense staining).

Results. In normal kidney, CD40 was expressed on parietal epithelial cells, mesangial cells, endothelial cells, and distal tubules but not proximal tubules. Glomerular and tubular CD40 expression was markedly up‐regulated in class III and class IV lupus GN, where there was intense staining of crescents, proximal and distal tubules, and interstitial mononuclear cells. In contrast, CD40 expression in class V lupus GN was similar to that in normal kidney. Interstitial mono‐nuclear cells expressing CD40L were present in class IV lupus GN. However, these findings were not unique to lupus GN: up‐regulation of CD40 and CD40L expression was similarly observed in other inflammatory renal diseases.

Conclusion. This study shows that CD40 is expressed on a variety of renal parenchymal and non‐parenchymal cells in normal kidney. Renal CD40 expression is up‐regulated in class III and class IV lupus nephritis, as well as in other inflammatory renal diseases, and is associated with the presence of CD40L+ mononuclear cells.
Suppression of type II collagen‐induced arthritis by the endogenous estrogen metabolite 2‐methoxyestradiol
Authors: Josefsson Elisabet , Tarkowski Andrej
Published: February 2007

Objective. To evaluate the antiarthritic properties of 2‐methoxyestradiol, an endogenous metabolite of estradiol, on type II collagen‐induced arthritis (CIA) in DBA/1 mice.

Methods. The effects of treatment with 2‐methoxyestradiol on the development of CIA were evaluated clinically and histologically. The in vitro effects of 2‐methoxyestradiol on lymphocyte and endothelial cell proliferation and differentiation were analyzed by standard methods.

Results. The development of CIA was significantly suppressed by 2‐methoxyestradiol. Incubation with 2‐methoxyestradiol suppressed the in vitro proliferation of endothelial cells, indicating that this compound down‐regulates angiogenesis. Endothelial cell production of nitric oxide (NO) was also down‐regulated by 2‐methoxyestradiol. In contrast to estradiol, 2‐methoxyestradiol exerted neither detectable feminizing effects on the sex organs nor inhibition of leukocyte development in hematopoietic organs.

Conclusion. The development of CIA is suppressed by 2‐methoxyestradiol, possibly via inhibition of angiogenesis. Diminished NO production could be of importance in vivo because it is a potent proinflammatory mediator. Since 2‐methoxyestradiol exerts only mild side effects compared with estradiol, it is an interesting candidate for therapeutic use in inflammatory diseases.
Analysis of the synovial cell infiltrate in early rheumatoid synovial tissue in relation to local disease activity
Authors: Tak Paul P. , Smeets Tom J. M. , Daha Mohamed R. , Kluin Philip M. , Meijers Katharina A. E. , Brand Ronald , Meinders A. Edo , Breedveld Ferdinand C.
Published: February 2007

Objective. To define variations in the cellular infiltrate and in the expression of monokines in synovial tissue (ST) from rheumatoid arthritis (RA) patients with different durations of disease and different levels of disease activity.

Methods. The immunohistologic features of synovial biopsy specimens from 31 patients with early RA (<1 year) and 35 patients with longstanding RA (>5 years) were compared. The possible associations between these features and local disease activity, as measured by the score for pain in the biopsied knee joint, were also evaluated.

Results. The immunohistologic features were not dependent on disease duration. We found a positive correlation between the scores for knee pain and the semiquantitative scores for the number of macrophages, as well as the expression of interleukin‐6 and tumor necrosis factor α, whereas the correlation with the scores for CD4+ T cells was negative. Multivariate analysis showed that these correlations were highly statistically significant (P < 0.003).

Conclusion. The results do not support the view that inflammatory mechanisms in the synovial tissues of RA patients differ between early and late stages of the disease. The findings presented here are consistent with the concept that early RA is the result of a synovitis process of longer duration and that macrophage‐derived cytokines play an important role in maintaining the clinical signs of inflammation.
Role of interleukin‐4 and interleukin‐10 in murine collagen‐induced arthritis. Protective effect of interleukin‐4 and interleukin‐10 treatment on cartilage destruction
Authors: Joosten Leo A. B. , Lubberts Erik , Durez Patrick , Helsen Monique M. A. , Jacobs Marike J. M. , Goldman Michel , van den Berg Wim B.
Published: February 2007

Objective. To examine the role of endogenous interleukin‐4 (IL‐4) and interleukin‐10 (IL‐10) and the therapeutic effect of the addition of IL‐4 and IL‐10 in early and established murine collagen‐induced arthritis (CIA).

Methods. Murine recombinant IL‐4, IL‐10, or the combination was given intraperitoneally twice daily from the day of arthritis onset up to 7–10 days of CIA in DBA/1 mice. Anti‐IL‐4, anti–IL‐10, or both antibodies were given intraperitoneally before or after the onset of CIA. The effect of cytokine or anticytokine treatment was monitored visually by macroscopic scoring. Histology and reverse transcription‐polymerase chain reaction (RT‐PCR) analyses were performed at the end of the treatment period.

Results. IL‐4 alone did not provoke any effect, IL‐10 slightly suppressed the arthritis, but a more pronounced amelioration was found with the combination. This cooperative effect was noted after early treatment but also occurred when the start of treatment was delayed until 1 week after onset. Apart from suppression of macroscopic signs of inflammation, combined treatment with IL‐4/IL‐10 also reduced cellular infiltrates in the synovial tissue and caused pronounced protection against cartilage destruction. Moreover, levels of mRNA for tumor necrosis factor α (TNFα) and IL‐1 were highly suppressed both in the synovial tissue and in the articular cartilage. In contrast, levels of IL‐1 receptor antagonist (IL‐1Ra) mRNA remained elevated, which suggests that the mechanism of protection may be related to suppressed production of TNFα and IL‐1, with concomitant up‐regulation of the IL‐1Ra/IL‐1 balance. However, accelerated onset of CIA and increased severity could be achieved with neutralizing anti–IL‐10 antibodies. This expression could be further optimized with a combination of anti–IL‐4 and anti–IL‐10 antibodies, although anti–IL‐4 alone was without effect.

Conclusion. Our data are consistent with a dominant role of IL‐10 in the natural suppression of arthritis expression, whereas combined treatment with IL‐4 and IL‐10 appears of potential therapeutic value, not only at the onset, but also in established arthritis.
Growth factor regulation of chondrocyte integrins. Differential effects of insulin‐like growth factor 1 and transforming growth factor β on α1β1 integrin expression and chondrocyte adhesion to type VI collagen
Authors: Loeser Richard F.
Published: February 2007

Objective. The ability of growth factors to modulate integrin expression may be important with regard to processes involved in tissue repair and remodeling. This study was undertaken to determine the effect of transforming growth factor β (TGFβ) and insulin‐like growth factor 1 (IGF‐1) on chondrocyte β1 integrin expression and integrin‐mediated adhesion to extracellular matrix proteins.

Methods. Chondrocytes obtained from normal bovine articular cartilage were cultured in the presence or absence of 10% fetal bovine serum, 100 pM IGF‐1, or 100 pM TGF β. Integrin expression and function were measured by protein blotting of immunoprecipitated integrins, Northern blot analysis, and cell adhesion assays.

Results. Immunoprecipitation with an anti–β1 integrin antibody coprecipitated the α1 integrin subunit and a band representing α3 and α5, as previously reported. Compared with serum‐free cultures, the use of serum resulted in an average 10‐fold increase in the α1 band and a 12‐fold increase in the α3/α5 band. IGF‐1 increased α1 and α3/α5 by an average of 3‐fold and 4‐fold, respectively. TGFβ also increased α3/α5 by >5‐fold but decreased α1 to an average of 24% of that found in serum‐free controls. Northern blot analysis revealed that TGFβ significantly increased α5 integrin subunit RNA levels. IGF‐1 did not have a significant effect on α5 or α1 integrin subunit RNA levels, suggesting that its effects on integrins are posttranscriptional. In cell adhesion assays, TGFβ treatment resulted in a 50% decrease in the adhesion of chondrocytes to type VI collagen, while adhesion to type II collagen and fibronectin was stimulated. IGF‐1 stimulated adhesion to all 3 proteins. An α1 integrin blocking antibody inhibited up to 75% of the adhesion of human chondrocytes to type VI collagen.

Conclusion. Both IGF‐1 and TGFβ stimulate chondrocyte cell surface expression of the α3/α5 integrin subunit band and stimulate adhesion of chondrocytes to fibronectin and type II collagen. The 2 growth factors have opposite effects on expression of α1β1, with IGF‐1 increasing and TGFβ decreasing cell surface levels of this integrin. TGFβ‐treated cells also have decreased adhesion to type VI collagen. The opposing effects of IGF‐1 and TGFβ on chondrocyte expression of α1/β1 and on adhesion to type VI collagen suggest that α1β1 mediates chondrocyte adhesion to type VI collagen. This was confirmed by using an antibody to the α1 integrin subunit to block adhesion of chondrocytes to type VI collagen.