bluuebunny/crossref_metadata_2025 huggingface dataset:
Epstein‐Barr virus strain type and latent membrane protein 1 gene deletions in lymphomas in patients with rheumatic diseases
Authors: Natkunam Yasodha , Elenitoba‐Johnson Kojo S. , Kingma Douglas W. , Kamel Onsi W.
Published: February 2007

Objective. Recent studies have shown that immunomodulatory therapy for the treatment of rheumatic diseases can be associated with the development of Epstein‐Barr virus (EBV)‐associated lymphoproliferative disorders. The present study was undertaken to determine the strain type of EBV in lymphoproliferative disorders that occur in patients with rheumatic disease and to investigate EBV latent membrane protein 1 (LMP‐1) gene deletions that occur in these lymphoproliferative disorders.

Methods. Ten EBV‐associated lymphoid neoplasms in patients with rheumatoid arthritis or dermatomyositis were analyzed by polymerase chain reaction to determine EBV strain type and to investigate for the presence of a previously characterized 30‐basepair deletion in the LMP‐1 gene.

Results. The results indicated that lymphoproliferative disorders in these patients can harbor EBV strain type A or B, with a predominance of type A infection (80%). It was also shown that both wild‐type and mutated LMP‐1 genes can be found in these neoplasms, with the deleted form of the LMP‐1 gene occurring in one‐third of cases in this series.

Conclusion. LMP‐1 deletions associated with certain aggressive lymphoid neoplasms are not required for the genesis of lymphoproliferative disorders in patients with rheumatic disease. The relative frequencies of type A and type B EBV strains in these lymphoproliferative disorders show similarities to the frequencies in patients with post‐solid organ transplantation immunosuppression‐associated lymphoproliferative disorders.
Small fiber neuropathy and vasculitis
Authors: Lacomis David , Giuliani Michael J. , Steen Virginia , Powell Henry C.
Published: February 2007

Vasculitis‐associated neuropathy usually presents as multiple mononeuropathies or sensorimotor polyneuropathies that affect large nerve fibers; painful small fiber sensory neuropathy has not previously been described in association with vasculitis. This report describes 2 patients with small fiber neuropathy in whom vasculitis was found to be present. Patients with small fiber neuropathy for which no other cause has been identified should be evaluated for the presence of vasculitis.
Allelic markers close to prolactin are associated with HLA‐DRB1 susceptibility alleles among women with rheumatoid arthritis and systemic lupus erythematosus
Authors: Brennan Paul , Hajeer Ali , Ong Kai Ren , Worthington Jane , John Sally , Thomson Wendy , Silman Alan , Ollier Bill
Published: February 2007

Objective. To investigate linkage disequilibrium between HLA‐DRB1 disease susceptibility alleles and microsatellite markers close to the prolactin gene, among women with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) and normal controls.

Methods. DNA from 89 women with RA, 76 women with SLE, and 94 controls was typed for HLA‐DRB1 status and D6S422 and D6S285, 2 highly polymorphic microsatellite markers close to the prolactin gene. RA patients were stratified by DRB1*0401 status, and SLE patients were stratified by *0301 status.

Results. There was an excess frequency of D6S422*1 among SLE patients with DRB1*0301 (odds ratio [OR] 3.1). The frequency of this allele was also slightly in excess among RA patients with DRB1*0401 (OR 1.9). D6S285*5 was also in excess among female RA patients with DRB1*0401 (OR 3.5), and was slightly increased among female SLE patients with DRB1*0301. None of these alleles were found to be increased among *0401‐positive or *0301‐positive controls.

Conclusion. These data indicate that there may be linkage disequilibrium between HLA‐DRB1 alleles and microsatellite marker alleles close to the prolactin gene among women with RA and SLE. This suggests the possibility of extended haplotypes encoding for HLA‐DRB1 susceptibility and high prolactin production, which contribute to susceptibility to both RA and SLE.
Matrix metalloproteinase 13 (collagenase 3) in human rheumatoid synovium
Authors: Lindy Otso , Konttinen Yrjö T. , Sorsa Timo , Ding Yanli , Santavirta Seppo , Ceponis Arnoldas , López‐otín Carlos
Published: February 2007

Objective. To show the eventual presence and extent of production of matrix metalloproteinase 13 (MMP‐13, or collagenase 3) in rheumatoid synovial tissue samples and extracts, and to assess the inhibition characteristics of recombinant MMP‐13.

Methods. Immunohistochemical avidin‐biotin‐peroxidase complex staining/morphometry was used to analyze MMP‐13‐positive cells in situ. Neutral salt extraction of synovial tissue, electrophoresis of the extract in different buffer systems, and Western blotting were also used. The inhibitory properties of doxycycline, clodronate, pamidronate, and D‐penicillamine for recombinant enzyme were determined with a soluble type II collagen assay.

Results. MMP‐13 was detected in fibroblast‐ and macrophage‐like mononuclear cells in the synovial lining and stroma and in vascular endothelial cells. The overall expression of MMP‐13 in these cells in the synovial stroma was high in rheumatoid arthritis (86 ± 12%) compared with osteoarthritis (17 ± 5%) patient samples (P = 0.0027). In a high‐pH native electrophoresis gel, immunoreactivity to anti‐MMP‐1 and anti‐MMP‐13 were clearly separated, with anti‐MMP‐13‐immunoreactive material migrating faster than anti‐MMP‐1‐immuno‐reactive material. Finally, in contrast to MMP‐1 and MMP‐8, MMP‐13 was found to be relatively resistant to the inhibitory effects of doxycycline and clodronate in vitro.

Conclusion. Due to its localization in synovial tissue, its substrate profile, increased expression, and relative resistance to known MMP inhibitors, MMP‐13 is suggested to play a major role in the pathogenesis of tissue destruction in rheumatoid arthritis.
The B cell repertoire of patients with rheumatoid arthritis. II. Increased frequencies of IgG+ and IgA+ B cells specific for mycobacterial heat‐shock protein 60 or human type II collagen in synovial fluid and tissue
Authors: Rudolphi Ulrike , Rzepka Rita , Batsford Stephen , Kaufmann Stefan H. E. , von der Mark Klaus , Peter Hans H. , Melchers Inga
Published: February 2007

Objective. A qualitative and quantitative analysis of the functional, antigen‐specific B cell receptor repertoire of patients with rheumatoid arthritis (RA) in synovial and peripheral compartments.

Methods. B cells were activated to grow and differentiate at high efficiency in vitro under limiting‐dilution conditions. Isotype and specificity of the secreted Ig were tested by enzyme‐linked immunosorbent assay.

Results. In contrast to peripheral B cells, most synovial B cells had already switched to IgG/IgA in vivo. The frequencies of B cells specifically recognizing foreign antigens were decreased within the synovial population, whereas the frequencies of B cells specific for type II collagen, mycobacterial heat‐shock protein 60 (hsp60), or IgG Fc fragments were significantly increased, revealing a negative correlation in terms of frequencies.

Conclusion. B cells specific for human type II collagen, hsp60, and IgG Fc fragments are produced and/or expanded locally within the affected joints of RA patients. Thus, the specific immune system is definitely involved in the local inflammatory and destructive processes.
Development of in vitro model systems for destructive joint diseases. Novel strategies for establishing inflammatory pannus
Authors: Schultz Olaf , Keyszer Gernot , Zacher Josef , Sittinger Michael , Burmester Gerd R.
Published: February 2007

Objective. To establish a novel 3‐dimensional (3‐D) in vitro model for the investigation of destructive processes in rheumatoid arthritis (RA).

Methods. Two distinct culture systems were developed, consisting of RA synovial membrane and articular cartilage explants or interactive RA synovial cell/chondrocyte cultures embedded in 3‐D fibrin matrices. The expression of proteolytic enzymes, chondrocyte matrix architecture, and matrix degradation parameters was analyzed by immunohistochemistry.

Results. Of 28 RA explant cultures, 16 displayed an invasion of synovial tissue into the cartilage explants, compared with 1 of 8 osteoarthritis explants. The expression of collagenase and vascular cell adhesion molecule 1 could be demonstrated at the cartilage‐pannus junction. Of 20 interactive cell cultures, 18 revealed invasive behavior and remained vital for extended periods of time.

Conclusion. The models presented allow us to study distinct aspects of destructive joint diseases under in vitro conditions that resemble human pathology. Moreover, our model is able to supplement animal experiments in basic research and drug testing.
Dysregulation of interleukin‐10 production in relatives of patients with systemic lupus erythematosus
Authors: Llorente Luis , Richaud‐Patin Yvonne , Couderc Jacques , Alarcon‐Segovia Donato , Ruiz‐Soto Rodrigo , Alcocer‐Castillejos Natasha , Alcocer‐Varela Jorge , Granados Julio , Bahena Susana , Galanaud Pierre , Emilie Dominique
Published: February 2007

Objective. To evaluate interleukin‐10 (IL‐10) production in relatives of patients with systemic lupus erythematosus (SLE).

Methods. Production of IL‐10 was evaluated in 13 families in which several members had SLE. The constitutive IL‐10 production in SLE patients (n = 16) was compared with that in healthy members of these multiplex families (n = 70), in 30 SLE patients who had no relatives with SLE, and in 46 healthy unrelated controls.

Results. The level of IL‐10 production did not differ between SLE patients who were members and those who were not members of multiplex families (mean ± SEM 4,384 ± 908 pg/ml and 4,709 ± 560 pg/ml, respectively), but was higher in both groups than in healthy unrelated controls (515 ± 88 pg/ml). The healthy members of the multiplex families constitutively produced large amounts of IL‐10 (3,080 ± 311 pg/ml; P < 0.001 compared with healthy unrelated controls). This high IL‐10 production was independent of age and sex, and was similar in first‐ and second‐degree relatives of SLE patients. The IL‐10 was produced both by monocytes and by a subpopulation of B lymphocytes in SLE patients and in their relatives.

Conclusion. The dysregulation of IL‐10 production previously identified in SLE patients is also present in healthy members of families with several cases of SLE, and it may contribute to the immunologic abnormalities affecting relatives of SLE patients.
Collagen crosslinks in fibromyalgia
Authors: Sprott Haiko , Müller Andreas , Heine Hartmut
Published: February 2007

Objective. To determine if abnormal collagen metabolism is a characteristic of fibromyalgia.

Methods. The diagnosis of fibromyalgia was made according to the American College of Rheumatology criteria. Skin biopsy samples were obtained from the trapezius region of 8 patients with fibromyalgia. Urine was collected under standardized conditions from 55 control subjects and 39 patients with fibromyalgia, and serum was obtained from 17 controls and 22 patients with fibromyalgia. Pyridinoline (Pyd), an indicator of connective tissue disease, and deoxypyridinoline (Dpyd), an indicator of bone degradation, both of which represent products of lysyl oxidase‐mediated crosslinking in collagen, were analyzed by ion‐paired and gradient high‐performance liquid chromatography (HPLC) methods with fluorescence detection. Levels of hydroxyproline (Hyp), a collagen turnover marker, were also measured. The findings were related to creatinine levels, and the Pyd:Dpyd ratio was determined.

Results. Highly ordered cuffs of collagen were observed around the terminal nerve fibers by electron microscopic examination of biopsy tissue from all 8 patients with fibromyalgia, but were not observed in any of the control skin samples. The Pyd:Dpyd ratios in the urine and serum and the Hyp levels in the urine were significantly lower in patients with fibromyalgia than in healthy controls.

Conclusion. Decreased levels of collagen cross‐linking in fibromyalgia may contribute to remodeling of the extracellular matrix and collagen deposition around the nerve fibers, and may contribute to the lower pain threshold at the tender points. Analysis of altered collagen metabolism either by histologic examination on biopsy, or preferably, by HPLC analysis of collagen metabolites in urine or serum may aid in understanding more about the pathogenesis of fibromyalgia.
Effects of self‐care education on the health status of inner‐city patients with osteoarthritis of the knee
Authors: Mazzuca Steven A. , Brandt Kenneth D. , Katz Barry P. , Chambers Mary , Byr Donna , Hanna Mark
Published: February 2007

Objective. To evaluate a concise program of self‐care education delivered by an arthritis nurse specialist as an adjunct to primary care for inner‐city patients with knee osteoarthritis (OA).

Methods. An attention‐controlled clinical trial; 211 inner‐city patients with knee OA were assigned arbitrarily to education (E) or attention‐control (AC) conditions. Group E received an individualized 30‐60‐minute educational intervention that emphasized non‐pharmacologic management of joint pain, preservation of function by problem‐solving, and practice of principles of joint protection. Brief telephone contacts 1 week and 4 weeks later monitored and reinforced new self‐care activities. Group AC viewed a 20‐minute standardized public education presentation on arthritis and received followup telephone calls (only to encourage continued participation in the study). Outcomes included the Health Assessment Questionnaire (HAQ) Disability and Discomfort Scales, 10‐cm visual analog scales measuring knee pain at rest and while walking, and the Quality of Well‐Being (QWB) scale. Assessments were made at baseline and at 4‐month intervals for 1 year.

Results. A total of 165 subjects (78%) completed all assessments. After control for baseline status, group E had significantly lower scores for disability and resting knee pain throughout the year of postintervention followup (P < 0.05 for both). Effects were somewhat discordant. By 12 months, functional benefits had begun to wane, while the effect on resting knee pain had grown. The overall effects of education on walking knee pain, overall joint pain (by HAQ), and general health status (by QWB) were not significant.

Conclusion. Self‐care education for inner‐city patients with knee OA, delivered as an adjunct to primary care, was found to result in notable preservation of function and control of resting knee pain. The magnitude of the observed effects compares well with those of more labor‐intensive and time‐consuming intervention models. However, more sustained preservation of function and consistent effects on pain may require prolonged, more proactive followup, either by the patient educator or by a trained clinical assistant dedicated to the task of supporting self‐care by patients with knee OA.
Analysis of human t cell and b cell responses against u small nuclear ribonucleoprotein 70‐kd, b, and d polypeptides among patients with systemic lupus erythematosus and mixed connective tissue disease
Authors: Holyst Margaret‐Mary , Hill Don L. , Hoch Sallie O. , Hoffman Robert W.
Published: February 2007

Objective. To analyze T and B cell reactivity with U small nuclear RNP (snRNP) 70‐kd, B, and D polypeptides among patients with connective tissue disease (CTD) and to examine the functional characteristics of snRNP‐reactive T cell clones.

Methods. We used an snRNP enzyme‐linked immunosorbent assay and immunoblotting to characterize antibodies in patients' sera. We used human recombinant fusion proteins 70 kd, B, and D to stimulate and clone snRNP‐reactive T cells from CTD patients. We analyzed the cell surface phenotype, antigenic specificity, and cytokine profiles of T cell clones.

Results. Patients showed T cell responsiveness to snRNP polypeptides that paralleled their autoantibody reactivities. A total of 256 clones were generated, and clones were identified which were specific for the 70‐kd, B, or D polypeptides. Clones expressed a T helper cell phenotype, and were found to produce substantial quantities of both interleukin‐4 (IL‐4) and interferon‐γ, and lesser quantities of IL‐2 and IL‐6.

Conclusion. These results show that CTD patients have clonable circulating snRNP‐reactive T cells that parallel the specificity of snRNP‐reactive antibodies in their sera. The snRNP‐reactive T cells exhibit a helper cell phenotype and produce cytokines which are important in B cell help and differentiation.
Is knee joint proprioception worse in the arthritic knee versus the unaffected knee in unilateral knee osteoarthritis?
Authors: Sharma Leena , Pai Yi‐Chung , Holtkamp Kelly , Rymer W. Zev
Published: February 2007

Objective. Neuromuscular joint protection requires proprioceptive input and motor output. Impairment of proprioception in knee osteoarthritis (OA) may contribute to, and/or result from, the disease. If this impairment was exclusively a local result of OA, a between‐knee difference would be expected in patients with unilateral OA (UOA). To explore causal directions, 2 hypotheses were tested: 1) proprioception is worse in UOA patients versus elderly controls; 2) proprioception is worse in the arthritic knee versus the unaffected knee in UOA patients.

Methods. Twenty‐eight UOA patients (Kellgren‐Lawrence grade ⩾2 in 1 knee and <2 in the other knee) and 29 elderly controls were enrolled. The unaffected knee of each UOA patient and both knees of the elderly controls were required to meet symptom, examination, and radiographic criteria. Proprioception (detection threshold of joint displacement after slow, passive, automated knee motion), body mass index, pain, functional status, range of motion, and laxity were measured.

Results. UOA patients had worse proprioception than did elderly controls, in either knee. A between‐knee difference was not found in UOA patients.

Conclusion. Impaired proprioception is not exclusively a local result of disease in knee OA. The relative importance of impaired proprioception in the development and progression of knee OA will require longitudinal study.
New‐onset juvenile dermatomyositis. Comparisons with a healthy cohort and children with juvenile rheumatoid arthritis
Authors: Pachman Lauren M. , Hayford Jennifer R. , Hochberg Marc C. , Pallansch Mark A. , Chung Ahn , Daugherty Claire D. , Athreya Balu H. , Bowyer Suzanne L. , Fink Chester W. , Gewanter Harry L. , Jerath Rita , Lang Bianca A. , Szer Ilona S. , Sinacore James , Christensen Mary L. , Dyer Alan R.
Published: February 2007

Objective. To determine, in a case‐control study, if patients with new‐onset juvenile dermatomyositis (juvenile DM) have increased symptoms prior to onset, exposure to certain environmental conditions, frequency of familial autoimmune diseases, or antibody titers, compared with 2 control groups.

Methods. A structured interview with the families of 80 children with juvenile DM, 40 children with juvenile rheumatoid arthritis (JRA), or 23 healthy children, from the same geographic area as the children with juvenile DM, was conducted. All children's sera were tested for antibody to Toxoplasma gondii, herpes simplex virus (HSV), or coxsackievirus B (CVB).

Results. A high proportion of children with juvenile DM had constitutional symptoms 3 months before the disease‐onset date (P = 0.013 versus control children). Children with JRA had more relatives with rheumatoid arthritis (P = 0.0001) and pernicious anemia (P = 0.003) than did children with juvenile DM or healthy children. Among children ⩽7 years of age, elevated enteroviral titers were more frequent in those with juvenile DM (81%) and in healthy controls (90%) than in those with JRA (64%), suggesting a common environmental exposure. Titers to T gondii, HSV, or CVB 1‐6 were normal.

Conclusion. Frequencies of familial autoimmune disease, exposure to environmental factors, or elevated antibody titers to T gondii, HSV, or CVB are not increased in juvenile DM. Children with juvenile DM do have symptoms of illness 3 months before the disease‐onset date, and young patients have elevated enteroviral titers, as do young geographic controls.
A prospective, longitudinal, multicenter study of service utilization and costs in fibromyalgia
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 study, for the first time, service utilization and costs in fibromyalgia, a prevalent syndrome associated with high levels of pain, functional disability, and emotional distress.

Methods. Five hundred thirty‐eight fibromyalgia patients from 6 rheumatology centers were enrolled in a 7‐year prospective study of fibromyalgia outcome. Patients were assessed every 6 months with validated, mailed questionnaires which included questions regarding fibromyalgia symptoms and severity, utilization of services, and work disability.

Results. Fibromyalgia patients averaged almost 10 outpatient medical visits per year, and when nontraditional treatments were considered, this number increased to ∼1 visit per month. Patients were hospitalized at a rate of 1 hospitalization every 3 years. In each 6‐month study period, patients used a mean of 2.7 fibromyalgia‐related drugs. Costs increased over the course of the study. The mean yearly per‐patient cost in 1996 dollars was $2,274. However, results were skewed by high utilizers, and many patients used few services and had limited costs. Total costs and utilization were independently associated with the number of selfreported comorbid or associated conditions, functional disability, and global disease severity. Compared with patients with other rheumatic disorders, those with fibromyalgia were more likely to have lifetime surgical interventions, including back or neck surgery, appendectomy, carpal tunnel surgery, gynecologic surgery, abdominal surgery, and tonsillectomy, and were more likely than other rheumatic disease patients to report comorbid or associated conditions. Almost 50% of hospitalizations occurring during the study were related to fibromyalgia‐associated symptoms.

Conclusion. The average yearly cost for service utilization among fibromyalgia patients is $2,274. Fibromyalgia patients have high lifetime and current rates of utilization of all types of medical services. They report more symptoms and comorbid or associated conditions than patients with other rheumatic conditions, and symptom reporting is linked to service utilization and, to a lesser extent, functional disability and global disease severity.
Rheumatoid arthritis and the risk of malignancy
Authors: Cibere Jolanda , Sibley John , Haga May
Published: February 2007

Objective. To determine the relative risks of malignancy and of site‐specific malignancies in patients with rheumatoid arthritis (RA).

Methods. In a prospective cohort study, 862 patients with RA (96% white) were enrolled from 1966 to 1974 and were followed up for up to 35 years (mean 17.4 years) at the University of Saskatchewan Rheumatic Disease Unit. All diagnoses of cancer were crossreferenced with the Provincial Cancer Registry. Expected cancer incidence rates were determined based on province of Saskatchewan population statistics matched to each study patient for age, sex, and calendar year. Standardized incidence ratios (SIRs) of the observedto‐expected cancer incidence and 95% confidence intervals (95% CI) were then calculated.

Results. A total of 136 cases of cancer were observed compared with 168 expected (SIR 0.80, P = 0.011 [95% CI 0.67–0.95]). The relative risk of colorectal malignancy was significantly reduced in the RA study population (SIR 0.52, P = 0.037 [95% CI 0.25–0.96]). A significant excess of leukemia was found (SIR 2.47, P = 0.026 [95% CI 1.12–4.69]), whereas the incidence rates for Hodgkin's disease and non‐Hodgkin's lymphoma and all other site‐specific malignancies were not found to be significantly different from general population rates.

Conclusion. In our cohort of RA patients, colorectal cancer was detected in only half the expected number of patients. This risk reduction may be related to long‐term nonsteroidal antiinflammatory drug (NSAID) use in RA, as has been suggested in several other studies of long‐term NSAID use. An increased risk of leukemia was confirmed. This may be due to the persistent immune stimulation associated with RA itself, since other potential explanatory factors for increased leukemia were not apparent. Despite the excess of hemopoietic malignancy and despite treatment of RA with potentially oncogenic agents, the overall risk of malignancy was reduced in this RA cohort.
H‐ras oncogene point mutations in arthritic synovium
Authors: Roivainen Anne , Jalava Jari , Pirilä Laura , Yli‐Jama Tuomas , Tiusanen Hannu , Toivanen Paavo
Published: February 2007

Objective. To examine mutational activation of ras proto‐oncogenes in synovial tissue from patients with rheumatoid arthritis (RA) compared with synovial specimens from patients with osteoarthritis (OA) or other arthropathies. Synovial samples from cadavers, without any signs of joint disease, were used as control material.

Methods. Using a combination of polymerase chain reaction (PCR) and automated sequencing of the amplified PCR product, regions around codons 12, 13, and 61 of the H‐, K‐, and N‐ras proto‐oncogenes were analyzed. Confirmation of mutations was based on restriction fragment length polymorphism analysis and/or oligonucleotide hybridization.

Results. Four (6%) of 72 patients with RA, 2 (13%) of 16 with OA, and 1 (8%) of 12 with other arthropathies harbored mutant H‐ras proto‐oncogenes, and were heterozygous at codon 13 for the GGT→GAT (Gly→Asp) change. An unexpected mutation was found in the H‐ras gene, in which a heterozygous GTG→ATG (Val→Met) mutation was observed over codon 14. The incidence for this mutation was 39% (28 of 72) in RA patients, 94% (15 of 16) in OA patients, and 42% (5 of 12) in patients with other arthropathies. All samples carrying the codon 13 mutation of H‐ras were also codon 14‐mutated, i.e., double mutations existed. Identical point mutations were also detected in a few synovial specimens obtained from cadavers (n = 8), including a single case of double mutation. All specimens showed normal K‐ and N‐ras loci.

Conclusion. Activation of proto‐oncogene H‐ras by point mutation in codons 13 and 14 occurred in the synovial tissue of patients with RA, OA, or other arthropathies, as well as, to some extent, in the control synovia, indicating that the phenomenon is not specific for RA. In codon 14, incidence of the H‐ras point mutation was highest in OA tissue. The possible significance of this codon 14‐mutated H‐ras gene needs to be clarified.
Collagenase‐3 (matrix metalloprotease 13) is preferentially localized in the deep layer of human arthritic cartilage in situ. In vitro mimicking effect by transforming growth factor β
Authors: Moldovan Florina , Pelletier Jean‐Pierre , Hambor John , Cloutier Jean‐Marie , Martel‐Pelletier Johanne
Published: February 2007

Objective. To examine, by immunohistochemistry, the localization and distribution of human collagenase‐3 in normal, osteoarthritis (OA), and rheumatoid arthritis (RA) cartilage, and to investigate the effects of interleukin‐1β (IL‐1β) and transforming growth factor β (TGFβ) on the synthesis and distribution of collagenase‐3.

Methods. Human cartilage specimens were obtained from tibial plateaus. In the first series of experiments, the OA specimens were excised from fibrillated and nonfibrillated areas of cartilage, and RA specimens were excised from lesional areas, including the cartilage‐pannus junction when present. In the second series, full strips of cartilage were processed for culture in the presence or absence of IL‐1β (100 units/ml) or TGFβ (150 ng/ml). Each specimen was processed for immunohistochemical analysis using a collagenase‐3 monoclonal antibody.

Results. The number of cells that stained for collagenase‐3 in normal cartilage was very low (±3%). In OA cartilage, the percentage increased dramatically, and no difference was found between fibrillated and nonfibrillated areas. A statistically significant increase in the percentage of cells staining for collagenase‐3 was found in the deep layer compared with the superficial layer. This finding was noted in both the fibrillated areas (mean ± SEM 58.4 ± 1.6% and 40.1 ± 3.9%, respectively; P < 0.007) and the nonfibrillated areas (55.4 ± 3.2% and 43.2 ± 2.7%; P < 0.01). Similarly, RA cartilage showed a statistically significant (P < 0.001) increase in the level of chondrocytes staining positive for collagenase‐3 in the deep layers (46.4 ± 4.1%) compared with the superficial layers (26.2 ± 3.4%). In these RA specimens, the numbers of positively staining chondrocytes were similar both close to and at a distance from the pannus junction. Both IL‐1β and TGFβ increased the number of chondrocytes producing collagenase‐3. Interestingly, in normal specimens, TGFβ had a predominant effect in the deep layers, while IL‐1β had a greater effect on the superficial layers.

Conclusion. This study demonstrates that, in situ, the increase in the level of chondrocytes synthesizing collagenase‐3 in arthritic cartilage is predominant in the deep layers. The results further indicate that TGFβ can up‐regulate the level of this enzyme and, in normal cartilage in vitro, can cause a mimicking of the in situ distribution observed in arthritic cartilage.
Testosterone suppresses anti‐DNA antibody production in peripheral blood mononuclear cells from patients with systemic lupus erythematosus
Authors: Kanda Naoko , Tsuchida Tetsuya , Tamaki Kunihiko
Published: February 2007

Objective. To study the in vitro effect of testosterone on anti‐DNA antibody production in peripheral blood mononuclear cells (PBMC) from patients with systemic lupus erythematosus (SLE) in order to elucidate its regulatory role in SLE.

Methods. PBMC from SLE patients were cultured with testosterone. IgG anti‐double‐stranded DNA (anti‐dsDNA) antibody, total IgG, and cytokine activity in the supernatants were measured by enzyme‐linked immunosorbent assay.

Results. Testosterone suppressed both IgG anti‐dsDNA antibody and total IgG production in PBMC from SLE patients. Antibody production in B cells was also suppressed by testosterone, although the magnitude of its effect on B cells was lower than that on PBMC. Interleukin‐6 (IL‐6) partially restored the testosterone‐induced decrease in antibody levels in PBMC. Testosterone reduced IL‐6 production in monocytes.

Conclusion. These results suggest that testosterone may directly suppress anti‐DNA antibody production in PBMC from SLE patients by inhibiting B cell hyperactivity and, indirectly, by down‐regulating IL‐6 production in monocytes. These results support the therapeutic effects of testosterone on SLE.
FAS/FAS ligand expression and induction of apoptosis in chondrocytes
Authors: Hashimoto Sanshiro , Setareh Morteza , Ochs Robert L. , Lotz Martin
Published: February 2007

Objective. To examine the expression of Fas/Fas ligand and the role of this ligand/receptor interaction in the regulation of apoptosis in normal human articular chondrocytes and in osteoarthritis (OA) cartilage.

Methods. Normal and OA human knee cartilage and cells isolated from these tissues were tested for Fas expression by flow cytometry. Induction of apoptosis by antibody to Fas was analyzed by DAPI staining and electron microscopy.

Results. Treatment of freshly isolated normal human articular chondrocytes with an agonistic Fas antibody induced apoptosis in a subpopulation (‐20%) of the cells. Apoptosis induced by anti‐Fas was not dependent on nitric oxide (NO), and anti‐Fas also did not induce NO production. Analysis of isolated cells demonstrated similar levels of Fas expression on normal and OA chondrocytes (28% and 32%, respectively). In normal articular cartilage, Fas‐positive cells were located mainly in the superficial and midzones. In contrast, in fibrillated OA cartilage, surface layers were partially absent and Fas‐expressing cells were also detected in the deeper layers. Fas ligand messenger RNA was not detectable in resting or activated normal or OA chondrocytes. Analysis by electron microscopy showed the nuclear and cytoplasmic changes typical of apoptosis in cultures treated with antibody to Fas.

Conclusion. A subpopulation of chondrocytes expresses Fas and is susceptible to Fas‐induced apoptosis. Fas‐mediated chondrocyte apoptosis may contribute to cartilage degradation in arthritis.
Posttranscriptional regulation of collagenase‐1 gene expression in synoviocytes by adenosine receptor stimulation
Authors: Boyle David L. , Han Zuoning , Rutter Joni L. , Brinckerhoff Constance E. , Firestein Gary S.
Published: February 2007

Objective. To characterize the transcriptional and posttranscriptional regulation of collagenase‐1 by adenosine receptor stimulation in interleukin‐1 (IL‐1)‐stimulated fibroblast‐like synoviocytes (FLS).

Methods. FLS were stimulated with IL‐1 and either the nonselective adenosine agonist 5′‐N‐ethylcarboxamidoadenosine (NECA) or the adenylate cyclase activator forskolin. Electrophoretic mobility shift assays were performed to determine AP‐1 and cAMP‐responsive element binding protein (CREB) activation. Transcriptional activation was determined by transfecting HS68 dermal fibroblasts with a collagenasechloramphenicol acetyltransferase construct. Finally, collagenase messenger RNA (mRNA) half‐life was determined by activating cells in the presence of IL‐1, IL‐1 + NECA, or IL‐1 + forskolin and culturing cells in the presence of actinomycin D.

Results. NECA and forskolin had no effect on AP‐1 activation, c‐jun or c‐fos gene expression, or CREB phosphorylation. IL‐1 markedly increased collagenase promoter activity, and neither NECA nor forskolin blocked this action. Studies of mRNA half‐life showed that both NECA and forskolin decreased the half‐life of collagenase mRNA in IL‐1‐stimulated FLS and HS68 cells.

Conclusion. The findings of this study demonstrate that NECA and forskolin decrease collagenase gene expression in FLS and dermal fibroblasts due to enhanced mRNA degradation.
A linkage study across the T cell receptor A and T cell receptor B loci in families with rheumatoid arthritis
Authors: Hall F. C. , Brown M. A. , Weeks D. E. , Walsh S. , Nicod A. , Butcher S. , Andrews L. J. , Wordsworth B. P.
Published: February 2007

Methods. A linkage study was performed in 184 RA families from the UK Arthritis and Rheumatism Council Repository, each containing at least 1 affected sibpair. The microsatellites D14S50, TCRA, and D14S64 spanning the TCRA locus and D7S509, Vβ6.7, and D7S688 spanning the TCRB locus were used as DNA markers. The subjects were genotyped using a semiautomated polymerase chain reaction‐based method. Twopoint and multipoint linkage analyses were performed.

Results. Nonparametric single‐marker likelihood odds (LOD) scores were 0.49 (P = 0.07) for D14S50, 0.65 (P = 0.04) for TCRA, 0.07 (P = 0.29) for D14S64, 0.01 (P = 0.43) for D7S509, 0.0 (P = 0.50) for Vβ6.7, and 0.0 (P = 0.50) for D7S688. By multipoint analysis, there was no evidence of linkage at TCRB (LOD score 0), and the maximum LOD score at the TCRA locus was 0.37 (at D14S50). The presence of a susceptibility locus (LOD score < −2.0) was excluded, with lambda ≥ 1.8 at TCRA and ≥ 1.4 at TCRB.

Conclusion. These linkage studies provide no significant evidence of a major germline‐encoded TCRA or TCRB component of susceptibility to RA.
Increased nitric oxide production accompanied by the up‐regulation of inducible nitric oxide synthase in vascular endothelium from patients with systemic lupus erythematosus
Authors: Belmont H. Michael , Levartovsky David , Goel Archana , Amin Ashok , Giorno Ralph , Rediske John , Skovron Mary Louise , Abramson Steven B.
Published: February 2007

Objective. To investigate whether systemic lupus erythematosus (SLE) is accompanied by increased serum nitrite levels, whether active compared with inactive disease is associated with greater nitric oxide (NO) production, and whether endothelial cells or keratinocytes serve as cellular sources of NO by virtue of their increased expression of either constitutive nitric oxide synthase (cNOS) or inducible NOS (iNOS).

Methods. Fifty‐one serum samples (46 from patients with SLE) were analyzed for NO production by measuring nitrite levels in a calorimetric assay. Skin biopsy samples from 21 SLE patients and 11 healthy volunteers were evaluated immunohistochemically, using monoclonal antibodies, for endothelial cell and keratinocyte cNOS and iNOS expression.

Results. Serum nitrite levels were significantly elevated in the 46 patients with SLE (mean ± SEM 37 ± 6 μM/liter) compared with controls (15 ± 7 μM/liter; P < 0.01), and were elevated in patients with active SLE compared with those with inactive disease (46 ± 7 μM/liter versus 30 ± 7 μM/liter; P < 0.01). Serum nitrite levels correlated with disease activity (r = 0.47, P = 0.04) and with levels of antibodies to doublestranded DNA (r = 0.35, P = 0.02). Endothelial cell expression of iNOS in SLE patients (mean ± SEM score 1.5 ± 0.2) was significantly greater compared with controls (0.6 ± 0.2; P < 0.01), and higher in patients with active disease compared with those with inactive SLE (1.7 ± 0.2 versus 1.2 ± 0.2; P < 0.01). Keratinocyte expression of iNOS was also significantly elevated in SLE patients (0.9 ± 0.1) compared with controls (0.4 ± 0.1; P < 0.001). With regard to expression of cNOS, there were no differences between patients with active SLE, those with inactive SLE, and normal controls in either the vascular endothelium or the keratinocytes.

Conclusion. NO production is increased in patients with SLE, and 2 potential sources of excessive NO are activated endothelial cells and keratinocytes via up‐regulated iNOS.
Susceptibility to ankylosing spondylitis in twins the role of genes, HLA, and the environment
Authors: Brown Matthew A. , Kennedy L. Gail , Macgregor Alex J. , Darke Chris , Duncan Emma , Shatford Jane L. , Taylor Andrew , Calin Andrei , Wordsworth Paul
Published: February 2007

Objective. To determine the relative effects of genetic and environmental factors in susceptibility to ankylosing spondylitis (AS).

Methods. Twins with AS were identified from the Royal National Hospital for Rheumatic Diseases database. Clinical and radiographic examinations were performed to establish diagnoses, and disease severity was assessed using a combination of validated scoring systems. HLA typing for HLA‐B27, HLA‐B60, and HLA‐DR1 was performed by polymerase chain reaction with sequence‐specific primers, and zygosity was assessed using microsatellite markers. Genetic and environmental variance components were assessed with the program Mx, using data from this and previous studies of twins with AS.

Results. Six of 8 monozygotic (MZ) twin pairs were disease concordant, compared with 4 of 15 B27‐positive dizygotic (DZ) twin pairs (27%) and 4 of 32 DZ twin pairs overall (12.5%). Nonsignificant increases in similarity with regard to age at disease onset and all of the disease severity scores assessed were noted in disease‐concordant MZ twins compared with concordant DZ twins. HLA‐B27 and B60 were associated with the disease in probands, and the rate of disease concordance was significantly increased among DZ twin pairs in which the co‐twin was positive for both B27 and DR1. Additive genetic effects were estimated to contribute 97% of the population variance.

Conclusion. Susceptibility to AS is largely genetically determined, and the environmental trigger for the disease is probably ubiquitous. HLA‐B27 accounts for a minority of the overall genetic susceptibility to AS.
Clinical, laboratory, radiographic, and histopathologic features of methotrexate‐associated lung injury in patients with rheumatoid arthritis. A multicenter study with literature review
Authors: Kremer Joel M. , Alarcón Graciela S. , Weinblatt Michael E. , Kaymakcian Mari V. , Macaluso Maurizio , Cannon Grant W. , Palmer William R. , Sundy John S. , St. Clair E. William , Alexander Ronald W. , Smith G. J. Walker , Axiotis Constantine A.
Published: February 2007

Objective. To describe the clinical, laboratory, radiologic, and histopathologic features of methotrexate (MTX)‐induced lung injury in a combined cohort of selected patients with rheumatoid arthritis (RA) and all cases reported in the English‐language literature.

Methods. Retrospective combined cohort review and abstraction from the medical literature. Case reports were obtained from 6 centers that had 4 or more cases of potential MTX lung injury per site. RA patients who were seen between 1981 and 1993 and who satisfied predetermined criteria for the presence of MTX lung injury were identified.

Results. Twenty‐seven patients satisfied the criteria for definite MTX lung injury, and 2 for probable MTX lung injury. Predominant clinical features of MTX lung injury included shortness of breath in 27 patients (93.1%), which was present for 23.5 ± 22.3 days (mean ± SD), cough in 24 (82.8%), present for 26.9 ± 28.5 days, and fever in 20 (69.0%), present for 10.4 ± 12.8 days. Five patients (17.2%) died, compared with 12 of 68 (17.6%) reported in the medical literature. Four of the 6 patients who were re‐treated with MTX after an initial pulmonary event developed recurrent lung toxicity, resulting in 2 deaths, compared with a recurrence rate of 3 of 6 in the literature.

Conclusion. MTX lung injury is most often a subacute process, in which symptoms are commonly present for several weeks before diagnosis. Approximately 50% of the cases are diagnosed within 32 weeks from initiation of MTX treatment. A patient who recovers from MTX lung injury should not be re‐treated. Earlier recognition and drug withdrawal may avoid the serious and sometimes fatal outcome that has been observed in this and other studies.
Combination treatment of severe rheumatoid arthritis with cyclosporine and methotrexate for forty‐eight weeks. An open‐label extension study
Authors: Stein C. Michael , Pincus Theodore , Yocum David , Tugwell Peter , Wells George , Gluck Oscar , Kraag Gunnar , Torley Helen , Tesser John , McKendry Robert , Brooks Raye H. ,   The Methotrexate‐Cyclosporine Combination Study Group
Published: February 2007

Objective. To determine whether the clinical benefit and favorable safety profile previously noted with the combination of cyclosporine (CSA) and methotrexate (MTX) given for 24 weeks in patients with rheumatoid arthritis (RA) would be maintained for a further 24 weeks, and whether the addition of CSA in patients who had previously been randomized to receive placebo + MTX would result in clinical benefit.

Methods. Eligible subjects from the initial study (weeks 0–24), in which the addition of placebo or CSA to MTX therapy was compared in patients with RA that was partially responsive to MTX, were enrolled. Patients who had received CSA + MTX continued this regimen for a further 24 weeks (weeks 24–48) (group 1; n = 48), and patients who had initially received placebo + MTX now received CSA + MTX for 24 weeks (weeks 24–48) (group 2; n = 44), in an open‐label extension study. The primary outcome measures were the number of tender joints, number of swollen joints, physician and patient global assessments, pain, functional disability as measured by the modified Health Assessment Questionnaire, and erythrocyte sedimentation rate.

Results. Of the 92 patients enrolled, 80 (87%) completed the extension study. In patients in group 1, the clinically and statistically significant improvement in response outcomes previously noted at week 24, ranging from 25% to 50%, was maintained through week 48. In patients in group 2, the addition of CSA resulted in significant clinical improvement. By week 48, most outcome measures in group 2 patients were similar to those in group 1 patients. CSA treatment resulted in a small increase in serum creatinine levels, but only 1 patient was withdrawn from the study for this reason.

Conclusion. The clinical improvement previously observed in patients treated with the CSA + MTX combination for 24 weeks was maintained for 24 subsequent weeks, without serious adverse effects, and was also observed in the patients whose treatment was switched from placebo + MTX to CSA + MTX.
Percutaneous needle muscle biopsy in the evaluation of patients with suspected inflammatory myopathy
Authors: Campellone Joseph V. , Lacomis David , Giuliani Michael J. , Oddis Chester V.
Published: February 2007

Objective. To determine the usefulness of a unique method of percutaneous needle muscle biopsy (NMB) in patients with suspected idiopathic inflammatory myopathy (IIM).

Methods. The yield of percutaneous NMB was studied in 55 patients who were found to have a combination of clinical, laboratory, or electromyographic features of IIM.

Results. A diagnosis of IIM was confirmed histopathologically in 29 patients (53%), other specific myopathies were found in 5 (9%), nonspecific myopathic changes were present in 11 (20%), and a neurogenic process was diagnosed in 3 (5%). Nonspecific changes or no abnormalities were present in 7 patients (13%). Followup of the 18 patients with nonspecific histopathologic findings disclosed that only 3 had a subsequent disease course compatible with IIM.

Conclusion. Percutaneous NMB is a safe, convenient, and relatively inexpensive method of muscle biopsy, with a high diagnostic yield for the pathologic confirmation of IIM. It should be considered as a primary method of acquiring muscle for histopathologic examination in the evaluation of suspected IIM.
Azathioprine‐induced severe pancytopenia due to a homozygous two‐point mutation of the thiopurine methyltransferase gene in a patient with juvenile HLA‐B27‐associated spondylarthritis
Authors: Leipold G. , Schütz E. , Haas J. P. , Oellerich M.
Published: February 2007

Severe pancytopenia due to azathioprine (AZA) toxicity in patients with autoimmune diseases is not uncommon. We describe a 14‐year‐old girl with HLA‐B27+ spondylarthritis who was treated with AZA 3 mg/kg/day and who suddenly developed severe pancytopenia in the seventh week of treatment. Analysis of the catabolic pathway of AZA revealed a homozygous deficiency of thiopurine methyltransferase (TPMT) on the basis of a combined 2‐point mutation at nucleotide positions 460 and 719 in the gene for TPMT, causing a toxic level of the metabolic active 6‐thioguanine nucleotides (6‐TGN) (2,394 pmoles/8 × 108 red blood cells). The patient was transfusion dependent and finally recovered 8 weeks after the development of the pancytopenia. At that time, 6‐TGN had already returned to normal therapeutic levels. Family studies revealed another homozygous deficiency in the mother, while the other family members were heterozygous.