bluuebunny/crossref_metadata_2025 huggingface dataset:
Important role of interleukin‐3 in the early phase of collagen‐induced arthritis
Authors: Brühl Hilke , Cihak Josef , Niedermeier Marianne , Denzel Andrea , Gomez Manuel Rodriguez , Talke Yvonne , Goebel Nicole , Plachý Jiří , Stangassinger Manfred , Mack Matthias
Published: April 2009

Objective

Activation of basophils contributes to memory immune responses and results in exacerbation of collagen‐induced arthritis (CIA). We undertook the present study to analyze the production and biologic effects of interleukin‐3 (IL‐3), a strong activator of basophils, in CIA.

Methods

Arthritis was induced by immunization with type II collagen. Mice were treated with blocking monoclonal antibodies against IL‐3 or with recombinant IL‐3. Clinical scoring, histologic analysis, fluorescence‐activated cell sorter analysis, enzyme‐linked immunosorbent assay, and cell culturing were performed to assess disease activity and IL‐3 production.

Results

IL‐3 was produced in large quantities by collagen‐specific CD4+ T cells in the spleen and was present in the synovial tissue during onset of arthritis, but was down‐regulated in paws with severe inflammation. Blockade of IL‐3 during the time of arthritis onset resulted in profound improvement of the disease, with reductions in synovial leukocyte and cytokine levels, peripheral blood basophil levels, and anticollagen antibody titers. Blockade of IL‐3 during the late phase of arthritis had no beneficial effect. Administration of recombinant IL‐3 during onset of arthritis induced a marked exacerbation of the disease, with increased peripheral blood basophil and plasma IL‐6 levels and increased titers of anticollagen antibody. In studies of the regulation of IL‐3 expression in CD4+ T cells, IL‐6 and IL‐4 suppressed the release of IL‐3 by activated CD4+ T cells, whereas lipopolysaccharide and CpG DNA up‐regulated IL‐3 secretion in activated CD4+ T cells by acting on costimulatory cells.

Conclusion

Taken together, the present results demonstrate for the first time that IL‐3 has an important role in the early phase of CIA.
Coexpression of CD177 and membrane proteinase 3 on neutrophils in antineutrophil cytoplasmic autoantibody–associated systemic vasculitis: Anti–proteinase 3–mediated neutrophil activation is independent of the role of CD177‐expressing neutrophils
Authors: Hu N. , Westra J. , Huitema M. G. , Bijl M. , Brouwer E. , Stegeman C. A. , Heeringa P. , Limburg P. C. , Kallenberg C. G. M.
Published: April 2009

Objective

Wegener's granulomatosis (WG) is strongly associated with antineutrophil cytoplasmic autoantibodies (ANCAs) directed against proteinase 3 (PR3). Recent studies have shown that membrane‐bound PR3 (mPR3) is differentially expressed and colocalizes with CD177/NB1 on circulating neutrophils. We undertook this study to assess the differential expression of CD177 on neutrophils from patients with ANCA‐associated systemic vasculitis (ASV) in comparison with patients with systemic lupus erythematosus (SLE), patients with rheumatoid arthritis (RA), and healthy individuals, and to investigate whether colocalization of mPR3 and CD177 affects anti‐PR3–mediated neutrophil activation.

Methods

Expression of CD177 and mPR3 was analyzed by flow cytometry on isolated neutrophils from patients with ASV (n = 53), those with SLE (n = 30), those with RA (n = 26), and healthy controls (n = 31). Neutrophil activation mediated by anti‐PR3 antibodies was assessed by measuring the oxidative burst with a dihydrorhodamine assay.

Results

Percentages of CD177‐expressing neutrophils were significantly higher in patients with ASV and those with SLE than in healthy controls. In 3 healthy donors, CD177 expression was not detected. After priming with tumor necrosis factor α, neutrophils remained negative for CD177 while mPR3 expression was induced. Neutrophils from CD177‐negative donors or CD177− neutrophils sorted from donors with bimodal expression were susceptible to anti‐PR3–mediated oxidative burst. Variation in the extent of anti‐PR3–mediated neutrophil activation among different donors occurred independent of the percentage of CD177‐expressing neutrophils.

Conclusion

Membrane expression of CD177 on circulating neutrophils is increased in patients with ASV and in those with SLE, but not in RA patients. However, primed neutrophils from CD177‐negative individuals also express mPR3 and are susceptible to anti‐PR3–mediated oxidative burst, suggesting that recruitment of CD177‐independent mPR3 is involved in anti‐PR3–induced neutrophil activation.
Cartilage repair in a rat model of osteoarthritis through intraarticular transplantation of muscle‐derived stem cells expressing bone morphogenetic protein 4 and soluble flt‐1
Authors: Matsumoto Tomoyuki , Cooper Gregory M. , Gharaibeh Burhan , Meszaros Laura B. , Li Guangheng , Usas Arvydas , Fu Freddie H. , Huard Johnny
Published: April 2009

Objective

The control of angiogenesis during chondrogenic differentiation is an important issue affecting the use of stem cells in cartilage repair, especially with regard to the persistence of regenerated cartilage. This study was undertaken to investigate the effect of vascular endothelial growth factor (VEGF) stimulation and the blocking of VEGF with its antagonist, soluble Flt‐1 (sFlt‐1), on the chondrogenesis of skeletal muscle‐derived stem cells (MDSCs) in a rat model of osteoarthritis (OA).

Methods

We investigated the effect of VEGF on cartilage repair in an immunodeficiency rat model of OA after intraarticular injection of murine MDSCs expressing bone morphogenetic protein 4 (BMP‐4) in combination with MDSCs expressing VEGF or sFlt‐1.

Results

In vivo, a combination of sFlt‐1– and BMP‐4–transduced MDSCs demonstrated better repair without osteophyte formation macroscopically and histologically following OA induction, when compared with the other groups. Higher differentiation/proliferation and lower levels of chondrocyte apoptosis were also observed in sFlt‐1– and BMP‐4–transduced MDSCs compared with a combination of VEGF‐ and BMP‐4–transduced MDSCs or with BMP‐4–transduced MDSCs alone. In vitro experiments with mixed pellet coculture of MDSCs and OA chondrocytes revealed that BMP‐4–transduced MDSCs produced the largest pellets, which had the highest gene expression of not only type II collagen and SOX9 but also type X collagen, suggesting formation of hypertrophic chondrocytes.

Conclusion

Our results demonstrate that MDSC‐based therapy involving sFlt‐1 and BMP‐4 repairs articular cartilage in OA mainly by having a beneficial effect on chondrogenesis by the donor and host cells as well as by preventing angiogenesis, which eventually prevents cartilage resorption, resulting in persistent cartilage regeneration and repair.
Prevention of glucocorticoid‐induced bone loss in mice by inhibition of RANKL
Authors: Hofbauer Lorenz C. , Zeitz Ute , Schoppet Michael , Skalicky Monika , Schüler Christiane , Stolina Marina , Kostenuik Paul J. , Erben Reinhold G.
Published: April 2009

Objective

RANKL has been implicated in the pathogenesis of glucocorticoid‐induced osteoporosis. This study was undertaken to evaluate the efficacy of denosumab, a neutralizing monoclonal antibody against human RANKL (hRANKL), in a murine model of glucocorticoid‐induced osteoporosis.

Methods

Eight‐month‐old male homozygous hRANKL‐knockin mice expressing a chimeric RANKL protein with a humanized exon 5 received 2.1 mg/kg of prednisolone or placebo daily over 4 weeks via subcutaneous slow‐release pellets and were additionally treated with phosphate buffered saline or denosumab (10 mg/kg subcutaneously twice weekly). Two groups of wild‐type mice were also treated with either prednisolone or vehicle.

Results

The 4‐week prednisolone treatment induced loss of vertebral and femoral volumetric bone mineral density in the hRANKL‐knockin mice. Glucocorticoid‐induced bone loss was associated with suppressed vertebral bone formation and increased bone resorption, as evidenced by increases in the number of tartrate‐resistant acid phosphatase (TRAP)–positive osteoclasts, TRAP‐5b protein in bone extracts, serum levels of TRAP‐5b, and urinary excretion of deoxypyridinoline. Denosumab prevented prednisolone‐induced bone loss by a pronounced antiresorptive effect. Biomechanical compression tests of lumbar vertebrae revealed a detrimental effect of prednisolone on bone strength that was prevented by denosumab.

Conclusion

Our findings indicate that RANKL inhibition by denosumab prevents glucocorticoid‐induced loss of bone mass and strength in hRANKL‐knockin mice.
The good initial response to therapy with a combination of traditional disease‐modifying antirheumatic drugs is sustained over time: The eleven‐year results of the Finnish rheumatoid arthritis combination therapy trial
Authors: Rantalaiho Vappu , Korpela Markku , Hannonen Pekka , Kautiainen Hannu , Järvenpää Salme , Leirisalo‐Repo Marjatta , Hakala Markku , Puolakka Kari , Julkunen Heikki , Luosujärvi Riitta , Möttönen Timo ,   FIN‐RACo Trial Group
Published: April 2009

Objective

To evaluate the evolution of functional and clinical outcomes over 11 years in patients with early rheumatoid arthritis (RA) initially treated with a combination of 3 disease‐modifying antirheumatic drugs (DMARDs) or with a single DMARD.

Methods

A cohort of 199 patients with early active RA were initially randomized to receive treatment with a combination of methotrexate, sulfasalazine, and hydroxychloroquine with prednisolone or treatment with a single DMARD (initially, sulfasalazine) with or without prednisolone. After 2 years, the drug treatment strategy became unrestricted, but still targeted remission. At 11 years, function was assessed with the Health Assessment Questionnaire (HAQ), and clinical outcomes were assessed with the modified Minimal Disease Activity (MDA) measure and the American College of Rheumatology (ACR) criteria for remission.

Results

At 11 years, 138 patients were assessed (68 in the combination‐DMARD group and 70 in the single‐DMARD group). The mean ± SD HAQ scores were 0.34 ± 0.54 in the combination‐DMARD group and 0.38 ± 0.58 in the single‐DMARD group (P = 0.88). Modified MDA was achieved by 63% (95% confidence interval [95% CI] 51, 77) and by 43% (95% CI 32, 55) (P = 0.016) of the combination‐DMARD group and the single‐DMARD group, respectively, and ACR remission by 37% (95% CI 26, 49) and by 19% (95% CI 11, 29) (P = 0.017), respectively.

Conclusion

Initial therapy with a combination of DMARDs in early RA results in higher rates of patients achieving modified MDA and strict ACR remission even over the long term than initial single‐DMARD therapy. Targeting remission with tight clinical controls results in good functional and clinical outcomes in most RA patients.
Induction of CCR2‐dependent macrophage accumulation by oxidized phospholipids in the air‐pouch model of inflammation
Authors: Kadl Alexandra , Galkina Elena , Leitinger Norbert
Published: April 2009

Objective

Macrophages are key players in the pathogenesis of rheumatoid synovitis as well as in atherosclerosis. To determine whether atherogenic oxidized phospholipids potentially contribute to synovial inflammation and subsequent monocyte/macrophage recruitment, we examined the effects of oxidized 1‐ palmitoyl‐2‐arachidonoyl‐sn‐3‐glycero‐phosphorylcholine (OxPAPC) on chemokine expression and leukocyte recruitment in a facsimile synovium in vivo using the murine air‐pouch model.

Methods

Air pouches were raised by 2 injections of sterile air, and inflammation was induced by injecting either lipopolysaccharide (LPS) or OxPAPC into the pouch lumen. Inflammation was assessed by analysis of inflammatory gene expression using reverse transcription–polymerase chain reaction or immunohistochemical analysis, and leukocytes were quantified in the lavage fluid and in the pouch wall after staining with Giemsa or after enzymatic digestion followed by fluorescence‐activated cell sorter analysis.

Results

Application of OxPAPC resulted in selective recruitment of monocyte/macrophages into the air‐pouch wall, but not in the lumen. In contrast, LPS induced both monocyte and neutrophil accumulation in the pouch lumen as well as in the wall. LPS, but not OxPAPC, induced the expression of adhesion molecules E‐selectin, P‐selectin, intercellular adhesion molecule 1, and vascular cell adhesion molecule 1. OxPAPC increased the expression of the CCR2 ligands monocyte chemotactic protein 1 (MCP‐1), MCP‐3, and MCP‐5, as well as RANTES and growth‐related oncogene α (GROα), while it down‐regulated the expression of CCR2 on macrophages. Moreover, oxidized phospholipid–induced macrophage accumulation was abrogated in CCR2−/− mice.

Conclusion

These data demonstrate that oxidized phospholipids trigger a type of inflammatory response that leads to selective macrophage accumulation in vivo, a process relevant for the pathogenesis of chronic inflammatory rheumatic diseases.
Activation of the interferon‐γ signaling pathway in systemic lupus erythematosus peripheral blood mononuclear cells
Authors: Karonitsch Thomas , Feierl Eva , Steiner Carl Walter , Dalwigk Karolina , Korb Adelheid , Binder Nikolaus , Rapp Alfred , Steiner Günter , Scheinecker Clemens , Smolen Josef , Aringer Martin
Published: April 2009

Objective

To investigate interferon‐γ (IFNγ) signaling in peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE) by analyzing IFNγ receptor (IFNγR) expression, STAT‐1 expression and phosphorylation, and the regulation of IFNγ‐inducible genes.

Methods

Fluorocytometry was used to investigate expression of STAT‐1, pSTAT‐1, CD95, HLA–DR, class I major histocompatibility complex (MHC), IFNγ‐inducible 10‐kd protein (IP‐10), monokine induced by IFNγ (Mig), and IFNγR in PBMCs from SLE patients and healthy individuals. STAT‐1 phosphorylation was determined by fluorocytometry and Western blotting after stimulation with IFNα or IFNγ. Quantitative polymerase chain reaction was used to assess messenger RNA (mRNA) expression of the IFNγ‐inducible genes IP‐10 and Mig shortly after preparation or after stimulation with IFNγ in monocytes.

Results

STAT‐1 expression was increased in PBMCs from SLE patients and correlated significantly with disease activity and with the IFN‐inducible expression of CD95 and HLA–DR. STAT‐1 expression also showed a trend toward association with class I MHC expression. In addition, the expression of other IFNγ‐inducible genes, such as IP‐10 or Mig, was increased in SLE monocytes. While STAT‐1 phosphorylation in SLE PBMCs and PBMCs from healthy individuals was similar after IFNα stimulation, incubation with IFNγ induced STAT‐1 phosphorylation only in SLE lymphocytes. Moreover, SLE monocytes showed a considerably higher increase in pSTAT‐1 expression upon IFNγ stimulation than monocytes from healthy individuals. Increased responsiveness of SLE monocytes to IFNγ was also confirmed on the mRNA level, where expression of the IFN‐inducible, STAT‐1–dependent genes IP‐10 and Mig was more efficiently increased in SLE cells. However, IFNγR was similarly expressed on SLE lymphocytes and monocytes and those from healthy individuals.

Conclusion

In addition to supporting the role of IFNs in SLE immunopathogenesis in general, the findings of the present study support a role of IFNγ in this disease.
Cadherin 11 promotes invasive behavior of fibroblast‐like synoviocytes
Authors: Kiener Hans P. , Niederreiter Birgit , Lee David M. , Jimenez‐Boj Esther , Smolen Josef S. , Brenner Michael B.
Published: April 2009

Objective

To define the expression pattern of cadherin 11 in the destructive pannus tissue of patients with rheumatoid arthritis, and to determine whether cadherin 11 expression in fibroblast‐like synoviocytes controls their invasive capacity.

Methods

Cadherin 11 expression in rheumatoid synovial tissue was evaluated using immunohistochemistry. To examine the role of cadherin 11 in regulating the invasive behavior of fibroblast‐like synoviocytes, we generated L cell clones expressing wild‐type cadherin 11, mutant cadherin 11, and empty vector–transfected controls. The invasive capacity of L cell transfectants and cultured fibroblast‐like synoviocytes treated with a blocking cadherin 11–Fc fusion protein or control immunoglobulin was determined in Matrigel invasion assays.

Results

Immunohistochemical analysis revealed that cadherin 11 is abundantly expressed in cells at the cartilage–pannus junction in rheumatoid synovitis. Assays to determine invasion demonstrated a 2‐fold increased invasive capacity of cadherin 11–transfected L cells compared with L cells transfected with E‐cadherin or control vector. The invasive behavior of L cells stably transfected with a cadherin 11 construct that lacked the juxtamembrane cytoplasmic domain was diminished to the level of vector control L cells. Furthermore, treatment with the cadherin 11–Fc fusion protein diminished the invasive capacity of fibroblast‐like synoviocytes.

Conclusion

The results of these in vitro studies implicate a role for cadherin 11 in promoting cell invasion and contribute insight into the invasive nature of fibroblast‐like synoviocytes in chronic synovitis and rheumatoid arthritis.
Comparison of drug retention rates and causes of drug discontinuation between anti–tumor necrosis factor agents in rheumatoid arthritis
Authors: Pan Sophie Martin Du , Dehler Silvia , Ciurea Adrian , Ziswiler Hans‐Rudolf , Gabay Cem , Finckh Axel
Published: April 2009

Objective

Tumor necrosis factor (TNF) inhibitors have revolutionized the treatment of severe rheumatoid arthritis (RA), yet drug discontinuation is common. The aim of this study was to compare treatment retention rates and specific causes of anti‐TNF discontinuation in a population‐based RA cohort.

Methods

All patients treated with etanercept, infliximab, or adalimumab within the Swiss Clinical Quality Management RA cohort between 1997 and 2006 were included in the study. Causes of treatment discontinuation were broadly categorized as adverse events (AEs) or nontoxic causes, and further subdivided into specific categories. Specific causes of treatment interruption were analyzed using a Cox proportional hazards model and adjusted for potential confounders.

Results

A total of 2,364 anti‐TNF treatment courses met the inclusion criteria. Treatment discontinuation was reported 803 times: 309 with etanercept, 249 with infliximab, and 245 with adalimumab. Drug inefficacy represented the largest single cause of treatment discontinuation (55.8% of cases). The median time of receiving anti‐TNF therapy was 37 months, but discontinuation rates differed between the 3 anti‐TNF agents (P < 0.001), with shorter retention rates for infliximab (hazard ratio [HR] 1.24, 99% confidence interval [99% CI] 1.01–1.51). The specific causes of treatment discontinuation revealed an increased risk of AEs with infliximab (HR 1.4, 99% CI 1.003–1.96), mostly due to an increased risk of infusion or allergic reactions (HR 2.11, 99% CI 1.23–3.62). Other discontinuation causes were equally distributed between the anti‐TNF agents.

Conclusion

In this population, infliximab was associated with higher overall discontinuation rates compared with etanercept and adalimumab, which is mainly due to an increased risk of infusion or allergic reactions.
Problems experienced by people with arthritis when using a computer
Authors: Baker Nancy A. , Rogers Joan C. , Rubinstein Elaine N. , Allaire Saralynn H. , Wasko Mary Chester
Published: April 2009

Objective

To describe the prevalence of computer use problems experienced by a sample of people with arthritis, and to determine differences in the magnitude of these problems among people with rheumatoid arthritis (RA), osteoarthritis (OA), and fibromyalgia (FM).

Methods

Subjects were recruited from the Arthritis Network Disease Registry and asked to complete a survey, the Computer Problems Survey, which was developed for this study. Descriptive statistics were calculated for the total sample and the 3 diagnostic subgroups. Ordinal regressions were used to determine differences between the diagnostic subgroups with respect to each equipment item while controlling for confounding demographic variables.

Results

A total of 359 respondents completed a survey. Of the 315 respondents who reported using a computer, 84% reported a problem with computer use attributed to their underlying disorder, and ∼77% reported some discomfort related to computer use. Equipment items most likely to account for problems and discomfort were the chair, keyboard, mouse, and monitor. Of the 3 subgroups, significantly more respondents with FM reported more severe discomfort, more problems, and greater limitations related to computer use than those with RA or OA for all 4 equipment items.

Conclusion

Computer use is significantly affected by arthritis. This could limit the ability of a person with arthritis to participate in work and home activities. Further study is warranted to delineate disease‐related limitations and develop interventions to reduce them.
Association of biomarkers with pre–radiographically defined and radiographically defined knee osteoarthritis in a population‐based study
Authors: Cibere Jolanda , Zhang Hongbin , Garnero Patrick , Poole A. Robin , Lobanok Tatiana , Saxne Tore , Kraus Virginia B. , Way Amanda , Thorne Anona , Wong Hubert , Singer Joel , Kopec Jacek , Guermazi Ali , Peterfy Charles , Nicolaou Savvakis , Munk Peter L. , Esdaile John M.
Published: April 2009

Objective

To evaluate 10 biomarkers in magnetic resonance imaging (MRI)–determined, pre–radiographically defined osteoarthritis (pre‐ROA) and radiographically defined OA (ROA) in a population‐based cohort of subjects with symptomatic knee pain.

Methods

Two hundred one white subjects with knee pain, ages 40–79 years, were classified into OA subgroups according to MRI‐based cartilage (MRC) scores (range 0–4) and Kellgren/Lawrence (K/L) grades of radiographic severity (range 0–4): no OA (MRC score 0, K/L grade <2), pre‐ROA (MRC score ≥1, K/L grade <2), or ROA (MRC score ≥1, K/L grade ≥2). Urine and serum samples were assessed for levels of the following biomarkers: urinary biomarkers C‐telopeptide of type II collagen (uCTX‐II), type II and types I and II collagen cleavage neoepitopes (uC2C and uC1,2C, respectively), and N‐telopeptide of type I collagen, and serum biomarkers sC1,2C, sC2C, C‐propeptide of type II procollagen (sCPII), chondroitin sulfate 846 epitope, cartilage oligomeric matrix protein, and hyaluronic acid. Multicategory logistic regression was performed to evaluate the association of OA subgroup with individual biomarker levels and biomarker ratios, adjusted for age, sex, and body mass index.

Results

The risk of ROA versus no OA increased with increasing levels of uCTX‐II (odds ratio [OR] 3.12, 95% confidence interval [95% CI] 1.35–7.21), uC2C (OR 2.13, 95% CI 1.04–4.37), and uC1,2C (OR 2.07, 95% CI 1.06–4.04), and was reduced in association with high levels of sCPII (OR 0.53, 95% CI 0.30–0.94). The risk of pre‐ROA versus no OA increased with increasing levels of uC2C (OR 2.06, 95% CI 1.05–4.01) and uC1,2C (OR 2.06, 95% CI 1.12–3.77). The ratios of type II collagen degradation markers to collagen synthesis markers were better than individual biomarkers at differentiating the OA subgroups, e.g., the ratio of [uCTX‐II][uC1,2C] to sCPII was associated with a risk of ROA versus no OA of 3.47 (95% CI 1.34–9.03) and a risk of pre‐ROA versus no OA of 2.56 (95% CI 1.03–6.40).

Conclusion

Different cartilage degradation markers are associated with pre‐ROA than are associated with ROA, indicating that their use as diagnostic markers depends on the stage of OA. Biomarker ratios contrasting cartilage degradation with cartilage synthesis are better able to differentiate OA stages compared with levels of the individual markers.
Efficacy, pharmacodynamics, and safety of VX‐702, a novel p38 MAPK inhibitor, in rheumatoid arthritis: Results of two randomized, double‐blind, placebo‐controlled clinical studies
Authors: Damjanov Nemanja , Kauffman Robert S. , Spencer‐Green George T.
Published: April 2009

Objective

To assess the efficacy and safety of VX‐702, a p38 MAPK inhibitor, in patients with active, moderate‐to‐severe rheumatoid arthritis (RA).

Methods

Two 12‐week, double‐blind, placebo‐controlled studies of VX‐702 were conducted in patients with active, moderate‐to‐severe RA. In the VeRA study, 313 patients received placebo or 2 daily doses of VX‐702. In Study 304, 117 patients received placebo, daily VX‐702, or twice weekly VX‐702 in addition to concomitant methotrexate (MTX). Study end points included the proportion of patients meeting the American College of Rheumatology 20% improvement criteria (an ACR20 response), ACR50 and ACR70 responses, changes in the serum levels of biomarkers of inflammation, and safety assessments.

Results

The numerically superior ACR20 response rates among patients receiving VX‐702 compared with those receiving placebo in both studies did not reach pairwise statistical significance at the highest doses in either study. At week 12 in the VeRA study, ACR20 response rates were 40%, 36%, and 28% among patients receiving 10 mg of VX‐702, 5 mg of VX‐702, and placebo, respectively. In Study 304, the response rates were 40%, 44%, and 22% for patients receiving 10 mg VX‐702 daily plus MTX, 10 mg VX‐702 twice weekly plus MTX, and placebo, respectively. Reductions in the levels of C‐reactive protein, soluble tumor necrosis factor receptor p55, and serum amyloid A were observed as early as week 1 in both studies, but these levels rapidly returned to baseline values by week 4. The overall frequency of adverse events was similar between the VX‐702 and placebo groups. In the VeRA study, serious infections were more frequent in the VX‐702 groups compared with the placebo group (2.4% versus 0%) but not in Study 304 (2.6% versus 4.9%).

Conclusion

The modest clinical efficacy plus the transient suppression of biomarkers of inflammation observed in this study suggest that p38 MAPK inhibition may not provide meaningful, sustained suppression of the chronic inflammation seen in RA.
Histone deacetylase 7, a potential target for the antifibrotic treatment of systemic sclerosis
Authors: Hemmatazad Hossein , Rodrigues Hanna Maciejewska , Maurer Britta , Brentano Fabia , Pileckyte Margarita , Distler Jörg H. W. , Gay Renate E. , Michel Beat A. , Gay Steffen , Huber Lars C. , Distler Oliver , Jüngel Astrid
Published: April 2009

Objective

We have recently shown a significant reduction in cytokine‐induced transcription of type I collagen and fibronectin in systemic sclerosis (SSc) skin fibroblasts upon treatment with trichostatin A (TSA). Moreover, in a mouse model of fibrosis, TSA prevented the dermal accumulation of extracellular matrix. The purpose of this study was to analyze the silencing of histone deacetylase 7 (HDAC‐7) as a possible mechanism by which TSA exerts its antifibrotic function.

Methods

Skin fibroblasts from patients with SSc were treated with TSA and/or transforming growth factor β. Expression of HDACs 1–11, extracellular matrix proteins, connective tissue growth factor (CTGF), and intercellular adhesion molecule 1 (ICAM‐1) was analyzed by real‐time polymerase chain reaction, Western blotting, and the Sircol collagen assay. HDAC‐7 was silenced using small interfering RNA.

Results

SSc fibroblasts did not show a specific pattern of expression of HDACs. TSA significantly inhibited the expression of HDAC‐7, whereas HDAC‐3 was up‐regulated. Silencing of HDAC‐7 decreased the constitutive and cytokine‐induced production of type I and type III collagen, but not fibronectin, as TSA had done. Most interestingly, TSA induced the expression of CTGF and ICAM‐1, while silencing of HDAC‐7 had no effect on their expression.

Conclusion

Silencing of HDAC‐7 appears to be not only as effective as TSA, but also a more specific target for the treatment of SSc, because it does not up‐regulate the expression of profibrotic molecules such as ICAM‐1 and CTGF. This observation may lead to the development of more specific and less toxic targeted therapies for SSc.
Transmembrane BAFF from rheumatoid synoviocytes requires interleukin‐6 to induce the expression of recombination‐activating gene in B lymphocytes
Authors: Rochas Caroline , Hillion Sophie , Saraux Alain , Mageed Rizgar A. , Youinou Pierre , Jamin Christophe , Devauchelle Valérie
Published: April 2009

Objective

B cells that accumulate in the synovial tissue of rheumatoid arthritis (RA) patients revise their receptors due to coordinate expression of recombination‐activating gene 1 (RAG‐1) and RAG‐2 genes. The aim of this study was to determine the mechanisms that control this re‐expression.

Methods

B cells from healthy control subjects were cocultured with fibroblast‐like synoviocytes (FLS) from patients with RA and osteoarthritis (OA). Re‐expression of RAG messenger RNA (mRNA) and proteins was analyzed by reverse transcription–polymerase chain reaction (RT‐PCR) and indirect immunofluorescence. Activity of RAG enzymes was evaluated by flow cytometry to measure variations in immunoglobulin κ and λ light chain expression and by ligation‐mediated–PCR to assess specific DNA breaks. Blocking antibodies, short hairpin RNA, and recombinant cytokine were used to identify the molecules involved in RAG re‐expression.

Results

RA FLS, but not OA FLS, induced B cells to re‐express RAG mRNA and proteins. Enzymes were functional, since the κ‐to‐λ ratios decreased and specific DNA breaks were detectable after coculture with RA FLS. Transmembrane BAFF provided the first signal of RAG re‐expression, since its down‐regulation in RA FLS prevented RAG gene transcription in B cells. The failure of transmembrane BAFF from OA FLS to induce RAG suggests that a second signal was provided by RA FLS. Interleukin‐6 (IL‐6) is a candidate, since blockade of its receptors precluded transcription of RAG genes by RA FLS. Unless supplemented with IL‐6, OA FLS were unable to induce RAG gene expression in normal B cells.

Conclusion

Two independent signals are required for the induction of RAG gene expression in B cells that infiltrate the synovium of patients with RA.
Th17 and natural Treg cell population dynamics in systemic lupus erythematosus
Authors: Yang Ji , Chu Yiwei , Yang Xue , Gao Di , Zhu Lubing , Yang Xinrong , Wan Linlin , Li Ming
Published: April 2009

Objective

To investigate the relative abundance and activities of Th17 cells and natural Treg cells in systemic lupus erythematosus (SLE).

Methods

Blood samples were collected from 50 adult patients with SLE. Samples were processed to detect Th17 cells and natural Treg cells by flow cytometry, and related gene expression was assessed by real‐time reverse transcription–polymerase chain reaction. Skin biopsy specimens were collected for histologic assessment. The function of Th17 cells in relation to human umbilical vein endothelial cells (HUVECs) was studied in vitro. Th17 cells were also examined in lupus‐prone MRL/Mp‐lpr/lpr (MRL/lpr) mice.

Results

We demonstrated the presence of Th17 cells among the peripheral blood mononuclear cells (PBMCs) and in the involved organs of patients with active SLE. Both the percentage of circulating Th17 cells and the ability to produce interleukin‐17A (IL‐17A) were increased in samples derived from patients with active SLE. The number of Th17 cells increased during SLE flare, especially in patients with vasculitis, and decreased following certain treatments. We observed that IL‐17A from patients with SLE could induce adhesion molecule messenger RNA expression in HUVECs and adhesion of T cells to HUVECs. An increase in the percentage of Th17 cells was correlated with natural Treg cell depletion during disease flare. Finally, expansion of the Th17 cell population was detected in MRL/lpr mice.

Conclusion

SLE flare might be linked to the expansion of the Th17 cell population and the depletion of natural Treg cell subpopulations. Expansion of the Th17 cell population might be related to a distinct cytokine environment in active SLE. Th17 cells and microenvironmental IL‐17A are involved in vascular inflammation in SLE. Antagonism of Th17 cells by IL‐17A–blocking antibodies should be explored as a treatment of SLE.
Development and validation of a composite disease activity score for juvenile idiopathic arthritis
Authors: Consolaro Alessandro , Ruperto Nicolino , Bazso Anna , Pistorio Angela , Magni‐Manzoni Silvia , Filocamo Giovanni , Malattia Clara , Viola Stefania , Martini Alberto , Ravelli Angelo
Published: April 2009

Objective

To develop and validate a composite disease activity score for juvenile idiopathic arthritis (JIA), the Juvenile Arthritis Disease Activity Score (JADAS).

Methods

The JADAS includes 4 measures: physician global assessment of disease activity, parent/patient global assessment of well‐being, active joint count, and erythrocyte sedimentation rate. These variables are part of the American College of Rheumatology (ACR) Pediatric 30 (Pedi 30), Pedi 50, and Pedi 70 criteria for improvement. Validation analyses were conducted on >4,500 patients and included assessment of construct validity, discriminant validity, and responsiveness to change. Three versions of the JADAS were tested based on 71‐joint (range 0–101), 27‐joint (range 0–57), or 10‐joint (range 0–40) counts. Statistical performances of the JADAS were compared with those of 2 rheumatoid arthritis composite scores, the Disease Activity Score in 28 joints (DAS28) and the Clinical Disease Activity Index (CDAI).

Results

The JADAS demonstrated good construct validity, yielding strong correlations with JIA activity measures not included in the score and moderate correlations with the Childhood Health Assessment Questionnaire. Correlations obtained for the 3 JADAS versions were comparable, but superior to those yielded by the DAS28 and CDAI. The area under the curve of the JADAS predicted long‐term disease outcome, measured as radiographic progression over 3 years. In 2 clinical trials, the JADAS discriminated well between ACR Pedi 30, Pedi 50, and Pedi 70 response and revealed strong responsiveness to clinical change.

Conclusion

The JADAS was found to be a valid instrument for assessment of disease activity in JIA and is potentially applicable in standard clinical care, observational studies, and clinical trials.
An adaptable search engine for multimodal information retrieval
Authors: Hubert Gilles , Mothe Josiane
Published: April 2009

This article describes an information retrieval approach according to the two different search modes that exist: browsing an ontology (via categories) or defining a query in free language (via keywords). Various proposals offer approaches adapted to one of these two modes. We present a proposal leading to a system allowing the integration of both modes using the same search engine. This engine is adapted according to each possible search mode.
Image‐based hysteresis modeling and compensation for an AFM piezo‐scanner
Authors: Zhang Yudong , Fang Yongchun , Zhou Xianwei , Dong Xiaokun
Published: April 2009

As an important component of Atomic Force Microscopes (AFM), a piezo‐scanner exhibits some undesired nonlinear characteristics, among which the inherent hysteresis largely decreases positioning accuracy during scanning and nano‐manipulation process. To alleviate this problem, an image‐based approach is proposed in this paper to model and then compensate for the hysteresis behavior of the piezo‐scanner. Specifically, some scanning images over standard samples are utilized to identify the parameters of the classical Preisach model (CPM) of hysteresis. On the basis of the obtained model, an inversion‐based technique is adopted to design a compensator for the hysteresis of the piezo‐scanner. The proposed algorithm presents such advantages as low cost and little complexity since no nanoscale position sensor is required to collect identification data. Some scanning and nano‐imprinting results are included to demonstrate the performance of the proposed strategy. Copyright © 2009 John Wiley and Sons Asia Pte Ltd and Chinese Automatic Control Society
Inhibitory effects of a gradient static magnetic field on normal angiogenesis
Authors: Wang Zhe , Yang Pengfei , Xu Huiyun , Qian Airong , Hu Lifang , Shang Peng
Published: April 2009

Angiogenesis, the formation of new blood vessels, is critical in many normal and pathological processes such as development, reproduction, tumor growth, and metastasis. Recently, exposure to moderate‐intensity static magnetic fields (1 mT to 1 T) has attracted much attention for its potential therapeutic value as a noninvasive intervening method. Nevertheless, the effects of moderate‐intensity and spatial gradient static magnetic fields (GSMF) on angiogenesis have not received enough attention. In this study, the effects of GSMF (0.2–0.4 T, 2.09 T/m, 1–11 days) on angiogenesis were investigated both in vitro and in vivo. An MTT assay was used as an in vitro method to detect the proliferation ability of human umbilical veins endothelial cells (HUVECs). Two kinds of in vivo models, a chick chorioallantoic membrane (CAM) and a matrigel plug, were used to detect the effects of GSMF on angiogenesis. The results showed that the proliferation ability of HUVECs was significantly inhibited 24 h after the onset of exposure. With regard to the CAM model, vascular numbers in the CAM that was continuously exposed to the GSMF were all less than those in normal condition. In accordance with the gross appearance, the contents of hemoglobin in the models exposed to GSMF for 7–9 days were also less. In addition, similar to the CAM model, the results of vascular density and hemoglobin contents in the matrigel plug also demonstrated that the GSMF exposure for 7 or 11 days inhibited vascularization. These findings indicate that GSMF might inhibit or prevent new blood vessels formation and could be helpful for the treatment of some diseases relevant to pathological angiogenesis. Bioelectromagnetics 30:446–453, 2009. © 2009 Wiley‐Liss, Inc.
Partition of thermodynamic energies of drug–DNA complexation
Authors: Kostjukov V. V. , Khomytova N. M. , Evstigneev M. P.
Published: April 2009

We report a computation methodology, which leads to the ability to partition the Gibb's free energy for the complexation reaction of aromatic drug molecules with DNA. Using this approach, it is now possible to calculate the absolute values of the energy contributions of various physical factors to the DNA binding process, whose summation gives a value that is reasonably close to the experimentally measured Gibb's free energy of binding. Application of the methodology to binding of various aromatic drugs with DNA provides an answer to the question “What forces are the main contributors to the stabilization of aromatic ligand–DNA complexes?” © 2009 Wiley Periodicals, Inc. Biopolymers 91: 773–790, 2009.

This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com
Randomized clinical trial of anal fistula plugversusendorectal advancement flap for the treatment of high cryptoglandular fistulain ano
Authors: Ortiz H , Marzo J , Ciga M A , Oteiza F , Armendáriz P , de Miguel M
Published: April 2009

Background

The aim of this randomized study was to compare the results of anal fistula plug and endorectal advancement flap in the treatment of high fistula in ano of cryptoglandular origin.

Methods

Consecutive patients with high trans-sphincteric fistula in ano of cryptoglandular aetiology were randomized to treatment with either an anal fistula plug or endorectal advancement flap. Patients agreed to participate in a follow-up programme, which included scheduled visits at 2, 4, 8, 12 and 24 weeks and at 1 year after surgery. The primary endpoint was effectiveness in fistula healing. Recurrence was defined as the presence of an abscess arising in the same area, or obvious evidence of fistulation.

Results

A large number of recurrences in the fistula plug group led to premature closure of the trial. After 1 year, fistula recurrence was noted in 12 of 15 patients treated with an anal fistula plug compared with two of 16 treated with an endorectal advancement flap (relative risk 6·40 (95 per cent confidence interval 1·70 to 23·97); P < 0·001).

Conclusion

Contrary to other published studies, an anal fistula plug was associated with a low rate of fistula healing, particularly in patients with a history of fistula surgery.
Quantitative separation of oxytocin, norfloxacin and diclofenac sodium in milk samples using capillary electrophoresis
Authors: Solangi Amber R. , Memon Saima Q. , Mallah Arfana , Khuhawar M. Y. , Bhanger M. I.
Published: April 2009

A simple, sensitive and rapid method has been developed for simultaneous separation and quantification of three different drugs: oxytocin (OT), norfloxacin (NOR) and diclofenac (DIC) sodium in milk samples using capillary electrophoresis (CE) with UV detection at 220 nm. Factors affecting the separation were pH, concentration of buffer and applied voltage. Separation was obtained in less than 9 min with sodium tetraborate buffer of pH 10.0 and applied voltage 30 kV. The separation was carried out from uncoated fused silica capillary with effective length of 50 cm with 75 µm i.d. The carrier electrolyte gave reproducible separation with calibration plots linear over 0.15–4.0 µg/mL for OT, 5–1000 µg/mL for NOR and 3–125 µg/mL for DIC. The lower limits of detection (LOD) were found to be 50 ng/mL for OT, and 1 µg/mL for NOR and DIC. The method was validated for the analysis of drugs in milk samples and pharmaceutical preparations with recovery of drugs within the range 96–100% with RSD 0.9–2.8%. Copyright © 2009 John Wiley & Sons, Ltd.
Analysis of hydroxyl radical‐induced oxidation process of glucagon by reversed‐phase HPLC and ESI‐MS/MS
Authors: Ichiba Hideaki , Ogawa Tomoko , Yajima Takehiko , Fukushima Takeshi
Published: April 2009

Structural modification of a polypeptide hormone, glucagon, by a hydroxyl radical in vitro was investigated by reversed‐phase high‐performance liquid chromatography (RP‐HPLC), and the oxidized site of glucagon was detected by electrospray ionization tandem mass spectrometry (ESI‐MS/MS). It was shown that 27methionine (Met) was oxidized to 27Met sulfoxide by hydroxyl radical, and the production rate of 27Met sulfoxide was faster than that by hydrogen peroxide. In addition, production of 27Met sulfoxide enantiomer was confirmed by RP‐HPLC analysis. cAMP production in a HepG2 cell induced by 27Met sulfoxide glucagon was reduced to approximately 75% as compared with that induced by the native form of glucagon. Copyright © 2009 John Wiley & Sons, Ltd.
Development and reproducibility of a novel high‐performance liquid‐chromatography monolithic column method for the detection and quantification of trans‐indolyl‐3‐acryloylglycine in human urine
Authors: Carr Kevin , Whiteley Paul , Shattock Paul
Published: April 2009

Elevated levels of trans‐indolyl‐3‐acryloylglycine (IAcrGly) have been reported in the urine of people with various conditions including pervasive developmental disorders (PDDs) such as autism and Asperger syndrome. Reversed‐phase high‐performance liquid chromatography with ultra‐violet detection using traditional particle silica‐based columns subsequent to solid‐phase extraction (SPE) has been the preferred assay method; requiring long analytical run times, high flow rates and high solvent usage. Recent developments in monolithic HPLC column technology facilitated the development of a novel analytical method, for the detection and quantification of urinary IAcrGly. The revised method eliminates the requirement for SPE pre‐treatment, reduces sample run‐time and decreases solvent volumes. Five urine samples from people diagnosed with PDD were run in quadruplicate to test the intra‐ and inter‐day reliability of the new method based on retention time, peak area and peak height for IAcrGly. Detection was by UV with IAcrGly confirmation by MS/MS‐MS. Relative standard deviations showed significant improvement with the new method for all parameters. The new method represents a major advancement in the detection and quantification of IAcrGly by reducing time and cost of analysis whilst improving detection limits and reproducibility. Copyright © 2009 John Wiley & Sons, Ltd.
On‐column refolding of corticotropin‐releasing factor receptor 1 extracellular domain by size exclusion chromatography
Authors: Ersoy Maria , Matic Sandra , Schmidt Stefan R.
Published: April 2009

Expressing the extracellular domain of corticotropin releasing factor receptor 1 in Escherichia coli usually results in the formation of inclusion bodies. Here we describe the optimization of refolding by applying size exclusion chromatography with a denaturing guanidine hydrochloride gradient and a refolding buffer containing glycerol. Several chromatographic parameters like gradient length, flow rate, sample concentration and chromatography resin characteristics were evaluated. Recovery yields of refolded protein above 50% using a Superdex 200 column demonstrate the usefulness of this method. Copyright © 2009 John Wiley & Sons, Ltd.
Association of a new intronic polymorphism of the SOD2 gene (G1677T) with cancer
Authors: Hernandez‐Saavedra Daniel , McCord Joe M.
Published: April 2009

There is growing evidence of the correlation between cancer and reactive oxygen species (ROS), especially superoxide. Low expression levels of the Mn‐superoxide dismutase (SOD2) enzyme have been reported in cancer patients. Genetic variation in the regulatory regions of the SOD2 gene may increase the risk of cancer. We identified a genetic variation (G1677T, rs2Y758Y339) in the vicinity of the enhancer region located in intron 2 of the SOD2 gene that creates a potential glucocorticoid responsive element, and developed an assay to screen DNA samples of 220 individuals (73 control, 59 prostate cancer survival individuals and 88 lung cancer biopsies). There were no significant differences in the genotype frequency distribution among prostate, lung cancer and control (p = 0.074 and 0.057, respectively). However, we identified an association of T allele with a decreased risk of lung cancer (OR = 0.525, p = 0.037). The use of the G1677T polymorphism of SOD2 gene as a genetic risk marker may suggest new approaches for detection, prevention, treatment, and prognosis of cancer. Copyright © 2009 John Wiley & Sons, Ltd.
Design of Triazole‐Tethered Glycoclusters Exhibiting Three Different Spatial Arrangements and Comparative Study of their Affinities towards PA‐IL and RCA 120 by Using a DNA‐Based Glycoarray
Authors: Moni Lisa , Pourceau Gwladys , Zhang Jing , Meyer Albert , Vidal Sébastien , Souteyrand Eliane , Dondoni Alessandro , Morvan François , Chevolot Yann , Vasseur Jean‐Jacques , Marra Alberto
Published: April 2009

Sugar‐coated chips: Glycoside clusters are valuable tools for carbohydrate–lectin recognition studies. However, the spatial arrangement of the sugar residues is a key issue in the design of high‐affinity glycoclusters. Here the affinities of linear and antenna‐ and calixarene‐based galactoside clusters towards two lectins derived from Pseudomonas aeruginosa and Ricinus communis were compared by means of glycoarrays.

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Interactions between proteins and carbohydrates are involved in a large number of crucial biological events. Many efforts have been devoted to the design and synthesis of unnatural saccharides displaying high affinities towards targeted lectins. Among others, glycoside clusters have proven to be valuable tools for these recognition studies. However, the spatial arrangements of the sugar residues are a key issue in the design of high‐affinity glycoclusters. Here, the affinities of linear and antenna‐ and calixarene‐based galactoside clusters against two lectins, derived from Pseudomonas aeruginosa and Ricinus communis, have been compared by means of glycoarrays.