bluuebunny/crossref_metadata_2025 huggingface dataset:
Adenosine A2A or A3 receptors are required for inhibition of inflammation by methotrexate and its analog MX‐68
Authors: Montesinos M. Carmen , Desai Avani , Delano Dave , Chen Jiang‐Fan , Fink J. Stephen , Jacobson Marlene A. , Cronstein Bruce N.
Published: January 2003

Objective

Low‐dose weekly methotrexate therapy remains a mainstay in the treatment of inflammatory arthritis. Results of previous studies demonstrated that adenosine, acting at one or more of its receptors, mediates the antiinflammatory effects of methotrexate in animal models of both acute and chronic inflammation. We therefore sought to establish which receptor(s) is involved in the modulation of acute inflammation by methotrexate and its nonpolyglutamated analog MX‐68 (N‐[[4‐[(2,4‐diaminopteridin‐6‐yl)methyl]‐3,4‐dihydro‐2H‐1,4‐benzothiazin‐7‐yl]‐carbonyl]‐L‐homoglutamic acid).

Methods

We studied the effects of low‐dose methotrexate (0.75 mg/kg intraperitoneally [IP] every week for 5 weeks), MX‐68 (2 mg/kg IP 2 days and 1 hour before induction of inflammation), dexamethasone (1.5 mg/kg IP 1 hour before induction of inflammation), or vehicle control on acute inflammation in an air‐pouch model in A2A and A3 receptor knockout mice.

Results

Low‐dose weekly methotrexate treatment increased the adenosine concentration in the exudates of all mice studied and reduced leukocyte and tumor necrosis factor α accumulation in the exudates of wild‐type mice, but not in those of A2A or A3 receptor knockout mice. Dexamethasone, an agent that suppresses inflammation by a different mechanism, was equally effective at suppressing leukocyte accumulation in A2A knockout, A3 knockout, and wild‐type mice, indicating that the lack of response was specific for methotrexate and MX‐68.

Conclusion

These findings confirm that adenosine, acting at A2A and A3 receptors, is a potent regulator of inflammation. Moreover, these results provide strong evidence that adenosine, acting at either or both of these receptors, mediates the antiinflammatory effects of methotrexate and its analog MX‐68.
Laser‐mediated microdissection facilitates analysis of area‐specific gene expression in rheumatoid synovium
Authors: Judex M. , Neumann E. , Lechner S. , Dietmaier W. , Ballhorn W. , Grifka J. , Gay S. , Schölmerich J. , Kullmann F. , Müller‐Ladner U.
Published: January 2003

Objective

Current approaches to analyzing gene expression in rheumatoid arthritis (RA) synovium are based on RNA isolated either from cultured synovial cells or from synovial biopsy specimens. This strategy does not, in general, allow distinction of specific gene expression between cells originating from different synovial areas, due to potential mixture of expression profiles. Therefore, we established the combination of laser‐mediated microdissection (LMM) and differential display to analyze profiles of gene expression in histologically defined areas of rheumatoid synovium. The present study was undertaken to establish parameters for this technique and assess its usefulness for gene expression analysis.

Methods

Cryosections derived from RA synovial tissues were used to obtain cell samples from synovial lining versus sublining, using a microbeam laser microscope. RNA was isolated and analyzed by nested RNA arbitrarily primed–polymerase chain reaction (RAP‐PCR) for differential display fingerprinting. Differentially expressed bands were cut out, and PCR products were eluted, cloned, and sequenced. Differential expression of identified sequences was confirmed by in situ hybridization and immunohistochemistry analysis.

Results

Microdissected sections of RA synovial tissue containing ∼600 cells yielded enough RNA to produce a reproducible RNA fingerprint pattern. Several genes could be identified as being expressed differentially between the synovial lining and the sublining, and their expression could be confirmed at the messenger RNA and protein levels.

Conclusion

The combination of LMM and RAP‐PCR presents a valuable tool to obtain novel insights into the area‐dependent differential regulation of gene expression in RA synovium. Both known and previously unknown genes were revealed with this technique. This study is the first to demonstrate the potential of this analytic strategy in the investigation of a nonmalignant, multifactorial, inflammatory disease.
The effect of HLA–DR on susceptibility to rheumatoid arthritis is influenced by the associated lymphotoxin α–tumor necrosis factor haplotype
Authors: Newton Julia , Brown Matthew A. , Milicic Anita , Ackerman Hans , Darke Chris , Wilson Jonathan N. , Wordsworth B. Paul , Kwiatkowski Dominic
Published: January 2003

Objective

HLA–DRB1, a major genetic determinant of susceptibility to rheumatoid arthritis (RA), is located within 1,000 kb of the gene encoding tumor necrosis factor (TNF). Because certain HLA–DRB1*04 subtypes increase susceptibility to RA, investigation of the role of the TNF gene is complicated by linkage disequilibrium (LD) between TNF and DRB1 alleles. By adequately controlling for this LD, we aimed to investigate the presence of additional major histocompatibility complex (MHC) susceptibility genes.

Methods

We identified 274 HLA–DRB1*04–positive cases of RA and 271 HLA–DRB1*04–positive population controls. Each subject was typed for 6 single‐nucleotide polymorphisms within a 4.5‐kb region encompassing TNF and lymphotoxin α (LTA). LTA–TNF haplotypes in these unrelated individuals were determined using a combination of family data and the PHASE software program.

Results

Significant differences in LTA–TNF haplotype frequencies were observed between different subtypes of HLA–DRB1*04. The LTA–TNF haplotypes observed were very restricted, with only 4 haplotypes constituting 81% of all haplotypes present. Among individuals carrying DRB1*0401, the LTA–TNF 2 haplotype was significantly underrepresented in cases compared with controls (odds ratio 0.5 [95% confidence interval 0.3–0.8], P = 0.007), while in those with DRB1*0404, the opposite effect was observed (P = 0.007).

Conclusion

These findings suggest that the MHC contains genetic elements outside the LTA–TNF region that modify the effect of HLA–DRB1 on susceptibility to RA.
Coordinate expression of activating Fcγ receptors I and III and inhibiting Fcγ receptor type II in the determination of joint inflammation and cartilage destruction during immune complex–mediated arthritis
Authors: Nabbe K. C. A. M. , Blom A. B. , Holthuysen A. E. M. , Boross P. , Roth J. , Verbeek S. , Van Lent P. L. E. M. , Van Den Berg W. B.
Published: January 2003

Objective

To study the role of the activating Fcγ receptor types I and III (FcγRI and FcγRIII, respectively) and the inhibiting Fcγ receptor II (FcγRII) in inflammation and in various aspects of cartilage destruction during arthritis that is solely induced by immune complexes.

Methods

Immune complex–mediated arthritis (ICA) was passively induced by lysozyme–antilysozyme complexes in FcγRI‐, FcγRIII‐, and FcγRII‐knockout mice and their wild‐type controls. Total knee joints were isolated to study inflammation and cartilage destruction (loss of proteoglycans [PGs], chondrocyte death, matrix metalloproteinase [MMP]–mediated neoepitope [VDIPEN] expression, and erosion). The presence of an active phenotype of macrophages was studied by detection of myeloid‐related proteins 8 and 14 (MRP8 and MRP14, respectively).

Results

Influx and activation of inflammatory cells (MRP expression) during ICA was decreased in FcγRIII‐deficient mice and enhanced in mice lacking FcγRII. Mild cartilage destruction reflected by loss of PGs was consistent with the degree of inflammation. Mice lacking FcγRIII showed almost no PG depletion, whereas in FcγRII−/− mice, PG depletion was increased 3–7‐fold in various cartilage areas. Initiation of erosive cartilage destruction, as reflected by MMP‐mediated VDIPEN expression, was reduced in FcγRIII−/− and FcγRI−/− mice, directing the two different critical steps of cellular influx and subsequent activation. These aspects were enhanced in FcγRII−/− mice. In FcγRI−/− and FcγRIII−/− mice, VDIPEN expression was 90–99% lower, whereas in FcγRII−/− mice, VDIPEN expression was increased 4‐fold. Chondrocyte death was reduced in FcγRIII−/− mice (68% lower) and enhanced in FcγRII−/− mice (6–12‐fold higher). Progression of arthritis and erosion of the cartilage surface were markedly elevated in FcγRII−/− arthritic joints.

Conclusion

During ICA, FcγRIII is the dominant activating receptor mediating joint inflammation, whereas both FcγRI and FcγRIII are involved in cartilage destruction. FcγRII inhibits both joint inflammation and severe cartilage destruction during ICA.
Induction of aggrecanase 1 (ADAM‐TS4) by interleukin‐1 occurs through activation of constitutively produced protein
Authors: Pratta Michael A. , Scherle Peggy A. , Yang Gengjie , Liu Rui‐Qin , Newton Robert C.
Published: January 2003

Objective

To study the production of aggrecanase 1 (ADAM‐TS4) in monolayer chondrocytes, capsular fibroblasts, and cartilage.

Methods

Bovine nasal and articular cartilage, monolayer chondrocytes, and capsular fibroblasts were incubated in the absence and presence of interleukin‐1 (IL‐1). ADAM‐TS4 production was evaluated by immunofluorescence or by Western blot analysis. Aggrecanase activity was measured in cells grown on an immobilized peptide substrate, and peptide cleavage was monitored by enzyme‐linked immunosorbent assay.

Results

There was constitutive production of ADAM‐TS4 in both cells and tissue. The protein was associated with the extracellular matrix based on the observation that the staining could be reduced following treatment of chondrocytes with heparin or exposure to chondroitinase ABC. Interestingly, there was no detectable change in the abundance of ADAM‐TS4 in response to IL‐1. Western blot analysis of cell lysates from IL‐1–stimulated chondrocytes showed no evidence of increased ADAM‐TS4 production, but resulted in activation of ADAM‐TS4. The activation was associated with an increased generation in the aggrecanase neoepitope NITEGE in nasal cartilage in response to IL‐1. These data suggest that induction of aggrecanase activity both in cells and in cartilage by IL‐1 may involve the stimulation of an activator of ADAM‐TS4. Consistent with this observation, culture of chondrocytes on a solid support containing a peptide substrate resulted in the generation of aggrecanase‐mediated cleavage that could be blocked by selective inhibitors of ADAM‐TS4.

Conclusion

These data support the hypothesis that ADAM‐TS4 is constitutively produced in these cells and tissue, and that stimulation by IL‐1 results in aggrecanase activation. Thus, the activator could be a potential target by which to control aggrecanase‐mediated degradation in arthritic diseases.
Influence of disease‐modifying therapy on radiographic outcome in inflammatory polyarthritis at five years: Results from a large observational inception study
Authors: Bukhari M. A. S. , Wiles N. J. , Lunt M. , Harrison B. J. , Scott D. G. I. , Symmons D. P. M. , Silman A. J.
Published: January 2003

Objective

To determine the effect of early treatment with disease‐modifying antirheumatic drugs (DMARDs) in reducing radiographic progression over a 5‐year period in patients with new‐onset inflammatory polyarthritis.

Methods

Three hundred thirty‐five consecutive patients with paired radiographs obtained 1 year and 5 years after enrollment in a population‐based arthritis register were studied. Logistic regression was used to model differences in baseline factors associated with the start of DMARDs. The time from symptom onset to first use of DMARDs was stratified to represent 4 groups: no DMARD use, <6 months, 6–12 months, and >12 months. Radiographs of the hands and feet were scored using the Larsen method. Progression in the Larsen score was evaluated as a 5‐year score adjusted for the first film score. Negative binomial regression was used to compare Larsen score progression for each of the 3 treatment groups with that for patients not receiving DMARDs. Results were then adjusted for severity, based on propensity modeling.

Results

Patients who received treatment had more radiographic progression than did patients who were untreated. Coefficients (95% confidence intervals), expressed as a multiple of the Larsen score in DMARD‐treated patients compared with untreated patients, were as follows: 1.6 (1.1–2.3) for <6 months, 2.4 (1.5–3.6) for 6–12 months, and 2.0 (1.4–2.8) for >12 months. As expected, patients receiving treatment had more severe disease at baseline. Using the propensity score as a method of adjusting for disease severity, the influence of treatment on outcome became attenuated as follows: 1.1 (0.8–1.7) for <6 months, 1.6 (1.0–2.6) for 6–12 months, and 1.5 (1.0–2.2) for >12 months. This effect was also seen in the crude Larsen score at year 5.

Conclusion

In this observational study, DMARD treatment was a marker not only of worse disease at presentation but also of the radiographic state and radiographic progression at 5 years. After adjustments were made for baseline disease severity, earlier therapy was shown to have a beneficial effect on outcome.
Human scleroderma sera contain autoantibodies to protein components specific to the U3 small nucleolar RNP complex
Authors: Yang Jian‐Ming , Hildebrandt Bernhard , Luderschmidt Christoph , Pollard K. Michael
Published: January 2003

Objective

To determine whether antifibrillarin autoantibodies in scleroderma patients are associated with autoantibodies to protein components specific for U3 small nucleolar RNP (U3 snoRNP).

Methods

Sera from 220 scleroderma patients were examined for antinucleolar autoantibodies (ANoA) and for antibodies to fibrillarin and the U3 snoRNP–specific proteins Mpp10 and hU3‐55K. Clinical correlates were determined for the different autoantibody specificities.

Results

Fifty‐nine of the 220 patients were positive for ANoA, and 31 of these patients were antifibrillarin positive. Anti–hU3‐55K was found in 10 patients, all of whom were antifibrillarin positive. Twenty‐nine patients had anti‐Mpp10 antibodies; 23 of these were antifibrillarin positive and 6 were antifibrillarin negative. ANoA, including antifibrillarin, anti–hU3‐55K, and anti‐Mpp10, were associated with diffuse, rather than limited, systemic or localized scleroderma. Esophageal and lung involvement were more common in patients with antifibrillarin and anti‐Mpp10 antibodies, and the highest frequency was in patients with anti‐Mpp10 alone.

Conclusion

Antifibrillarin autoantibodies are associated with autoantibodies to protein components specific to U3 snoRNP, particularly Mpp10. The prevalence of anti‐Mpp10 antibodies in antifibrillarin‐positive patients suggests that the U3 snoRNP particle is a source of immunogenic/antigenic material for the anti‐snoRNP response in scleroderma. Autoantibodies to snoRNP components were more frequent in patients with diffuse scleroderma than in those with either the limited systemic or localized forms. The increased expression of these antibodies in patients with the more severe form of scleroderma, coupled with the observations that fibrillarin expression is positively linked to collagen expression in fibroblasts and that fibrillarin is overexpressed in scleroderma fibroblasts, suggests a source of snoRNP to initiate and maintain these autoantibody responses.
Identification of an IL1A gene segment that determines aberrant constitutive expression of interleukin‐1α in systemic sclerosis
Authors: Kawaguchi Yasushi , Hara Masako , Kamatani Naoyuki , Wright Timothy M.
Published: January 2003

Objective

To further explore the mechanisms of transcriptional regulation of the IL1A gene in fibroblasts from the affected skin of patients with systemic sclerosis (SSc).

Methods

The transcription activity of IL1A was estimated by nuclear run‐on assay. A reporter construct (pLuc.–1437) was prepared that contained a 1.5‐kb genomic fragment extending from −1437 to +39 relative to the transcription start site of the IL1A gene inserted upstream of the luciferase gene. The DNA protein binding activity for the cis‐element of the IL1A gene was determined by electrophoretic mobility shift assay (EMSA).

Results

Affected fibroblasts exhibited a strong nuclear run‐on signal, indicative of constitutive transcription activity of the IL1A gene. Luciferase assay indicated high levels of transcription activity of pLuc.–1437 in transiently transfected fibroblasts from affected skin; this was absent in transiently transfected control fibroblasts. Analysis of constructs containing a series of 5′ deletions of pLuc.–1437 revealed that the sequence between −96 and −82 (nuclear factor–interleukin‐1α [NF–IL‐1α]) was the most important for transcription of the IL1A gene in affected SSc fibroblasts. NF–IL‐1α contains a 5′‐GGAA‐3′ sequence that is essential for specific binding of members of the Ets nuclear protein family. Using EMSA, a DNA protein complex was detected in samples containing nuclear extracts from affected fibroblasts and the NF–IL‐1α probe, whereas no specific band was detected in samples from normal fibroblasts. The DNA protein complex disappeared with the use of NF–IL‐1α probes that had mutations of the Ets binding sites.

Conclusion

These findings suggest that a DNA binding protein containing the Ets domain is constitutively expressed only in fibroblasts from the affected skin of SSc patients and regulates the transcription of the IL1A gene, which contributes to the fibrogenic phenotype of fibroblasts in SSc.
Functional disability in rheumatoid arthritis patients compared with a community population in Finland
Authors: Sokka Tuulikki , Krishnan Eswar , Häkkinen Arja , Hannonen Pekka
Published: January 2003

Objective

To compare Health Assessment Questionnaire (HAQ) scores of patients with rheumatoid arthritis (RA) with HAQ scores from a sex‐ and age‐adjusted population.

Methods

Patients with RA (n = 1,095) and control subjects (n = 1,530) completed a mailed questionnaire that comprised the HAQ, pain and global health scores, education level, and comorbidities, as well as height, weight, and lifestyle attitudes, including smoking and exercise habits.

Results

The HAQ scores increased (indicating declining function) with older age in patients and controls. The HAQ scores were above the reference values (>95th percentile of the HAQ scores of the age‐ and sex‐matched population) in 17–45% of women with RA and in 7–32% of men with RA ages 30–79 years, while the HAQ scores of the patients ≥80 years were similar to those of the age‐ and sex‐matched population. In a logistic regression model, the odds ratio for disability (HAQ score ≥1; at least some difficulties in most activities of daily living) was 7.7 (95% confidence interval 5.3–11.1; P < 0.001) among patients with RA compared with community controls, when adjusted for age, sex, education, smoking, exercise, body mass index, number of comorbidities, and pain.

Conclusion

RA is associated with a >7‐fold risk of disability compared with that in a general population of adults in the same community. The impact of disability due to RA appears to be greater in younger and middle‐age people than in elderly patients.
Control of fibroblast‐like synoviocyte proliferation by macrophage migration inhibitory factor
Authors: Lacey Derek , Sampey Annaleise , Mitchell Robert , Bucala Richard , Santos Lanie , Leech Michelle , Morand Eric
Published: January 2003

Objective

The hyperplasia of fibroblast‐like synoviocytes (FLS) is considered essential to the evolution of joint destruction in rheumatoid arthritis (RA), but the mechanisms underlying FLS proliferation remain poorly understood. Macrophage migration inhibitory factor (MIF) is a cytokine that has recently been shown to exert proinflammatory effects on RA FLS. This study sought to identify the mechanisms of activation of FLS by MIF, and to assess the effects of MIF on synovial cell proliferation.

Methods

Human RA FLS were treated with recombinant MIF, interleukin‐1β (IL‐1β), tumor necrosis factor α (TNFα), and/or anti‐MIF monoclonal antibodies (mAb). Proliferation was measured with tritiated thymidine incorporation. Nuclear factor κB (NF‐κB) and mitogen‐activated protein (MAP) kinase activation were measured with immunohistochemistry and Western blotting, respectively.

Results

FLS proliferation was significantly increased by MIF. IL‐1β and TNFα also induced proliferation, but these effects were prevented by neutralization with anti‐MIF mAb. Activation of NF‐κB was induced by IL‐1β, but not by MIF. Anti‐MIF mAb had no effect on IL‐1β–induced NF‐κB nuclear translocation. By contrast, MIF induced phosphorylation of extracellular signal–regulated kinase (ERK) MAP kinase. ERK antagonism, but not NF‐κB antagonism, prevented the effect of MIF on FLS proliferation.

Conclusion

These data suggest that MIF may regulate RA synovial hyperplasia by acting directly and via involvement in the effects of IL‐1β and TNFα. In addition, the effects of MIF on FLS activation are independent of NF‐κB, and dependent on ERK MAP kinase. These data suggest an important therapeutic potential for MIF antagonism in RA.
Increased salivary gland tissue expression of Fas, Fas ligand, cytotoxic T lymphocyte–associated antigen 4, and programmed cell death 1 in primary Sjögren's syndrome
Authors: Bolstad Anne Isine , Eiken Hans Geir , Rosenlund Benedikte , Alarcón‐Riquelme Marta E. , Jonsson Roland
Published: January 2003

Objective

To assess salivary gland tissues obtained from patients with primary Sjögren's syndrome (SS) for the gene expression profile of the candidate genes TNFRSF6 (Fas), TNFSF6 (FasL), SSA1 (Ro52α and the splice variant Ro52β), SSB (La), CTLA4, PDCD1 (PD‐1), and ORM2, which were selected on the basis of their putative participation in salivary gland inflammation.

Methods

Quantitative real‐time reverse transcriptase–polymerase chain reaction (RT‐PCR) was used to examine the expression of messenger RNA (mRNA). Tissue localization of the expressed proteins was detected by immunohistochemistry.

Results

Expression of mRNA was increased for Fas (5.1‐fold; P < 0.001), FasL (8.8‐fold; P < 0.05), cytotoxic T lymphocyte–associated antigen 4 (CTLA‐4) (11.2‐fold; P < 0.01), programmed cell death 1 (PD‐1) (15.2‐fold; P < 0.05), Ro52α (3.0‐fold; P < 0.01), La (2.3‐fold; P < 0.05), and orosomucoid 2 (ORM2) (4.4‐fold; P < 0.05) in patients compared with controls when GAPDH was used as endogenous standard in duplex runs. In single runs using 2 other endogenous standards (18S and β‐actin), statistically significant differences between patients and controls were confirmed for expression of Fas, FasL, CTLA‐4, and PD‐1, but this difference was not observed for Ro52α, La, and ORM2. Expression of Ro52β mRNA was similar in patients and controls.

Conclusion

The present study demonstrates a substantial increase in expression of the negative regulator molecules PD‐1 and CTLA‐4 and the apoptotic signal molecules Fas and FasL in patients with primary SS compared with controls, which corresponded to the immunomorphologic pattern. The results strongly indicate that these molecules have central roles in the inflammatory process in the salivary glands of patients with primary SS.
Screening for atherosclerosis in patients with rheumatoid arthritis: Comparison of two in vivo tests of vascular function
Authors: Van Doornum S. , McColl G. , Jenkins A. , Green D. J. , Wicks I. P.
Published: January 2003

Objective

Inflammation appears to play a central role in atherosclerosis, and endothelial damage mediated by systemic inflammation may contribute to the increased cardiovascular mortality in rheumatoid arthritis (RA). Brachial artery flow‐mediated dilatation (FMD) and pulse wave analysis (PWA) are measures of vascular function. The aim of this study was to determine if FMD and PWA are abnormal in patients with RA.

Methods

Twenty‐five RA patients and 25 matched healthy controls were studied. All were free of traditional cardiovascular risk factors. FMD was measured in all subjects. PWA was performed in 18 RA patients and 18 controls, with results expressed as large and small artery compliance (C1 and C2). Modified Sharp scores were calculated in 13 RA patients.

Results

Results (mean ± SD) in RA patients and controls, respectively, were as follows: FMD 107.6 ± 4.6% versus 108.5 ± 4.1% (P = 0.49), C1 14.8 ± 2.8 ml/mm Hg × 10 versus 17.9 ± 3.1 ml/mm Hg × 10 (P = 0.0033), C2 4.5 ± 2.3 ml/mm Hg × 100 versus 7.7 ± 3.7 ml/mm Hg × 100 (P = 0.0039). There was an inverse correlation between C2 and modified Sharp scores in the RA patients (Spearman's rho −0.69, P = 0.0085).

Conclusion

FMD was normal in these RA patients, whereas arterial compliance was markedly reduced. PWA appears to be a more sensitive measure of vascular dysfunction than FMD in RA and may be the preferred surrogate marker of vascular dysfunction in longitudinal studies of RA patients. The inverse correlation between C2 and the modified Sharp score, a measure that reflects disease activity over time, supports the notion that chronic inflammation plays a role in RA‐associated atherosclerosis.
Association of IL1A gene polymorphisms with susceptibility to and severity of systemic sclerosis in the Japanese population
Authors: Kawaguchi Yasushi , Tochimoto Akiko , Ichikawa Naomi , Harigai Masayoshi , Hara Masako , Kotake Shigeru , Kitamura Yutaka , Kamatani Naoyuki
Published: January 2003

Objective

To analyze the frequencies of haplotypes of single‐nucleotide polymorphisms (SNPs) of the IL1A gene (at −889, +4729, and +4845) in patients with systemic sclerosis (SSc) and in healthy control subjects, and to determine whether the IL1A gene haplotype is associated with SSc susceptibility or disease severity.

Methods

We studied 60 patients with SSc (34 with diffuse cutaneous SSc and 26 with limited cutaneous SSc) and 70 healthy control subjects. Polymorphisms of the IL1A gene were genotyped by direct sequencing using the ABI Prism 377 Sequence Detection System. The LDSupport program, which was recently developed in our laboratory, was used to estimate the haplotype frequencies of SNPs in the study population.

Results

We confirmed the presence of 2 SNPs at positions −889 (C/T) and +4845 (G/T) of the IL1A gene, as previously reported. We also identified a novel SNP at position +4729 (T/C). Six haplotypes, CTG (49.7%), TCT (14.7%), CCT (20.3%), TTG (13.2%), CCG (1.4%), and TTT (0.7%), were found in the healthy controls. In contrast, only 2 haplotypes, CTG (95%) and TCT (5%), were detected in the SSc patients. Notably, the CTG haplotype was present at a significantly higher frequency in the SSc patients than in the healthy controls (P < 0.0001). We also examined the relationship between the CTG/CTG diplotype frequencies and interstitial lung disease (ILD), a major complication of SSc, as an indicator of disease severity. All SSc patients with ILD had the CTG/CTG diplotype, whereas the frequency of this diplotype was only 67% in patients without ILD.

Conclusion

Our observations suggest that the CTG haplotype of the IL1A gene may be an important marker for the susceptibility to, and the severity of, SSc.
Serum amyloid P component binds to late apoptotic cells and mediates their uptake by monocyte‐derived macrophages
Authors: Bijl Marc , Horst Gerda , Bijzet Johan , Bootsma Hendrika , Limburg Pieter C. , Kallenberg Cees G. M.
Published: January 2003

Objective

Some pentraxins, such as C‐reactive protein, bind to apoptotic cells and are involved in the clearance of these cells. We undertook this study to determine whether serum amyloid P component (SAP; a pentraxin that, when deficient in mice, results in lupus‐like disease) binds to apoptotic cells and to assess the functional consequences of SAP binding for their phagocytosis by macrophages.

Methods

Human peripheral blood monocytes were isolated and cultured for 7 days to obtain monocyte‐derived macrophages. Jurkat cells were irradiated with ultraviolet B to induce apoptosis. After 4 hours, a mean ± SEM of 54.0 ± 5.1% of these cells stained with annexin V and were propidium iodide negative (early apoptotic [EA] cells). After 24 hours, 77.3 ± 2.7% of cells stained positive with both annexin V and propidium iodide (late apoptotic [LA] cells or secondary necrotic cells). EA and LA cells were incubated with fluorescein isothiocyanate–labeled SAP in the presence or absence of Ca2+, and binding was measured by flow cytometry. Phagocytosis was tested by incubation of macrophages with EA or LA cells in the presence of normal human serum (NHS) and quantified as a phagocytosis index (PI; number of Jurkat cells internalized by 100 macrophages). Experiments were repeated with SAP‐depleted serum and after reconstitution with increasing concentrations of SAP.

Results

The majority of LA cells did bind SAP in the presence of Ca2+, whereas EA cells did not. SAP depletion of NHS resulted in a 50% decrease in the PI for LA cells, and complete restoration of the PI could be demonstrated with SAP reconstitution up to 100 μg/ml. SAP depletion had no effect on phagocytosis of EA cells.

Conclusion

SAP binds to LA cells and is involved in the phagocytosis of these cells by human monocyte–derived macrophages. This may have consequences for diseases such as systemic lupus erythematosus, in which phagocytosis of apoptotic cells is decreased.
Elucidation of the relationship between synovitis and bone damage: A randomized magnetic resonance imaging study of individual joints in patients with early rheumatoid arthritis
Authors: Conaghan Philip G. , O'Connor Philip , McGonagle Dennis , Astin Paul , Wakefield Richard J. , Gibbon Wayne W. , Quinn Mark , Karim Zunaid , Green Michael J. , Proudman Susanna , Isaacs John , Emery Paul
Published: January 2003

Objective

To simultaneously image bone and synovium in the individual joints characteristically involved in early rheumatoid arthritis (RA).

Methods

Forty patients with early, untreated RA underwent gadolinium‐enhanced magnetic resonance imaging (MRI) of the second through fifth metacarpophalangeal joints of the dominant hand at presentation, 3 months, and 12 months. In the first phase (0–3 months), patients were randomized to receive either methotrexate alone (MTX) or MTX and intraarticular corticosteroids (MTX + IAST) into all joints with clinically active RA. The MTX‐alone group received no further corticosteroids until the second phase (3–12 months), when both groups received standard therapy.

Results

In the first phase, MTX + IAST reduced synovitis scores more than MTX alone. There were significantly fewer joints with new erosions on MRI in the former group compared with the latter. During the second phase, the synovitis scores were equivalent and a similar number of joints in each group showed new erosions on MRI. In both phases, there was a close correlation between the degree of synovitis and the number of new erosions, with the area under the curve for MRI synovitis the only significant predictor of bone damage progression. In individual joints, there was a threshold effect on new bone damage related to the level of synovitis; no erosions occurred in joints without synovitis.

Conclusion

In early RA, synovitis appears to be the primary abnormality, and bone damage occurs in proportion to the level of synovitis but not in its absence. In the treatment of patients with RA, outcome measures and therapies should focus on synovitis.
Reduced arterial elasticity in rheumatoid arthritis and the relationship to vascular disease risk factors and inflammation
Authors: Wong Melinda , Toh Ling , Wilson Andrew , Rowley Kevin , Karschimkus Connie , Prior David , Romas Evange , Clemens Laurence , Dragicevic George , Harianto Harry , Wicks Ian , McColl Geoff , Best James , Jenkins Alicia
Published: January 2003

Objective

Rheumatoid arthritis (RA) is associated with increased rates of cardiovascular disease. Reduced small artery elasticity (SAE) and large artery elasticity (LAE) and increased systemic vascular resistance (SVR) have been found in other high‐risk groups. In the present study, we sought to determine whether arterial elasticity was reduced and SVR was increased in RA patients compared with controls matched for coronary artery disease (CAD) status, and to relate the results to vascular disease risk factors, including measures of inflammation.

Methods

Arterial elasticity was assessed by pulse wave analysis in RA patients with (n = 15) and without (n = 38) CAD, and in controls matched 1:1 for age, sex, and CAD status. Vascular risk factors, including high‐sensitivity C‐reactive protein (hsCRP), soluble vascular cell adhesion molecule 1 (sVCAM‐1), and serum amyloid A (SAA) levels, were assessed.

Results

SAE and LAE were significantly lower and SVR was significantly higher in RA patients than in controls. RA patients also had higher levels of hsCRP, SAA, and sVCAM‐1. SAE and LAE values were inversely correlated with markers of inflammation. Associations of SAE and LAE with RA were independent of conventional risk factors, but were dependent on markers of inflammation.

Conclusion

Vascular function is abnormal in RA, with reduced SAE and LAE and increased SVR relative to controls. Arterial elasticity is inversely associated with measures of inflammation. These measures may be clinically useful in the detection and monitoring of vascular disease in RA.
High‐dose cyclophosphamide without stem cell transplantation in systemic lupus erythematosus
Authors: Petri Michelle , Jones Richard J. , Brodsky Robert A.
Published: January 2003

Objective

High‐dose chemotherapy followed by hematopoietic stem cell transplantation is increasingly being studied as a treatment for severe autoimmune disorders, such as systemic lupus erythematosus (SLE). High‐dose cyclophosphamide, the foundation of virtually all conditioning regimens for stem cell transplantation, is not myeloablative; therefore, when high‐dose cyclophosphamide is used alone, autografting, with its potential for reinfusing autoreactive effector cells, is not required. We undertook this study to investigate the safety and efficacy of high‐dose cyclophosphamide without stem cell transplantation in refractory SLE.

Methods

We treated 14 patients with moderate‐to‐severe SLE that had been refractory to corticosteroids and one or more additional immunosuppressive drugs. All patients received 50 mg/kg of cyclophosphamide for 4 consecutive days followed by 5 μg/kg granulocyte colony‐stimulating factor until the neutrophil count was 1 × 109/liter for 2 consecutive days. Patients were followed up monthly for disease activity using the physician's global assessment, SLE Disease Activity Index, and assessment of functioning of involved organs. The Responder Index for Lupus Erythematosus was used to define partial or complete response.

Results

The median time to achieve a neutrophil count of 0.5 × 109/liter was 14 days (range 11–22 days) after the last dose of cyclophosphamide. Patients required a median of 2 units (range 2–5) of packed red blood cells, and a median of 16 days (range 0–23 days) elapsed from the last dose of cyclophosphamide to the last platelet transfusion. There were no deaths or fungal infections. Significant improvements in physician's global assessment (mean difference 1.4; P < 0.0001), SLE Disease Activity Index (mean difference 4.1; P = 0.0039), and prednisone dosage (mean difference 14.9 mg/day; P = 0.002) were observed. Responses, including 5 durable complete responses, were observed in all organ systems (renal, central nervous system, and skin) with involvement that had led to patient enrollment.

Conclusion

High‐dose cyclophosphamide without stem cell transplantation leads to rapid hematopoietic reconstitution and has significant clinical benefit in patients with refractory SLE. Therefore, this approach deserves further study.
Jaccoud's arthropathy in systemic lupus erythematosus: Differentiation of deforming and erosive patterns by magnetic resonance imaging
Authors: Ostendorf Benedikt , Scherer Axel , Specker Christof , Mödder Ulrich , Schneider Matthias
Published: January 2003

Objective

To evaluate alterations of the soft tissues, tendons, and bones as detected by magnetic resonance imaging (MRI) in patients with systemic lupus erythematosus (SLE)–associated arthritis of the finger joints.

Methods

Both hands of 14 patients with SLE of various activities and durations and with arthritis and/or deformities of the finger joints were examined by MRI and conventional radiography. Coronal T1‐weighted spin‐echo (with and without gadolinium contrast), axial T2‐weighted turbo spin‐echo, coronal fat‐suppressed short tau inversion recovery, and 3‐dimensional double‐echo steady‐state sequences were acquired and analyzed, and the findings were compared with those from conventional radiographs.

Results

MRI detected periarticular capsular swelling in all 14 patients, joint effusion in 7, edematous tenosynovitis in 6, proliferative tenosynovitis (flexor and/or extensor tendons) in 4, and intraarticular signs of synovial membrane hypertrophy in 10 patients, 9 of whom showed enhancement after administration of contrast medium. Bony erosions were identified in 8 patients by MRI; conventional radiography missed these erosions in 2 of the 8 patients. Four of the 14 patients were designated as Jaccoud's arthropathy index–positive, and all 4 showed severe edematous tenosynovitis and capsular swelling, but no signs of bony erosions despite longstanding disease (mean 21.5 years). Four of the 10 patients with mild deformity exhibited prominent soft tissue pathology, with minimal destruction of bone; the other 6 patients had bony alterations that resembled rheumatoid arthritis.

Conclusion

In SLE patients with arthritis of the finger joints, MRI detects characteristic signs of soft tissue pathology (e.g., capsular swelling, edematous and proliferative tenosynovitis, synovial hypertrophy) and bony alterations (e.g., erosions, some of which are missed by conventional radiography). MRI thus helps to distinguish different types of lupus arthritis/Jaccoud's arthropathy, which allows more differentiated treatment strategies and monitoring.
Arthropathy in juvenile hemochromatosis
Authors: Vaiopoulos George , Papanikolaou George , Politou Marianna , Jibreel Iyad , Sakellaropoulos Nikos , Loukopoulos Dimitris
Published: January 2003

Objective

To evaluate whether arthropathy is associated with juvenile hemochromatosis and, if so, to assess the relationship between this feature and other clinical features of the disease.

Methods

Clinical, laboratory, and radiologic evidence of arthropathy was studied in 8 Greek patients with genetically proven juvenile hemochromatosis. Osteopenia and osteoporosis were assessed by bone mineral density measurement.

Results

Seven of the 8 patients had articular manifestations. The main affected joint was the metacarpophalangeal joint, and the arthritis was progressive independent of phlebotomy therapy. Osteopenia was observed in 2 patients, and osteoporosis in 2 patients.

Conclusion

Arthropathy may be present in patients with juvenile hemochromatosis, with features similar to those found in patients with hemochromatosis type 1.
A comparison between anti‐Th/To– and anticentromere antibody–positive systemic sclerosis patients with limited cutaneous involvement
Authors: Mitri Ghaith M. , Lucas Mary , Fertig Noreen , Steen Virginia D. , Medsger Thomas A.
Published: January 2003

Objective

To compare anti‐Th/To–positive and anticentromere antibody (ACA)–positive patients with limited cutaneous systemic sclerosis (lcSSc).

Methods

We reviewed the medical records of 107 anti‐Th/To–positive patients and 365 ACA‐positive patients who were first evaluated during 1985–2000. ACA was detected by indirect immunofluorescence on HEp‐2 cell substrate, and anti‐Th/To was detected by RNA immunoprecipitation with K562 cell extracts. Patients were included if they had a clinical diagnosis of lcSSc, and excluded if they had >1 SSc‐associated serum autoantibody.

Results

The final study groups comprised 87 lcSSc patients with anti‐Th/To antibodies and 306 with ACAs. Anti‐Th/To–positive patients were younger (P < 0.04) and had a shorter disease duration at their first evaluation (P < 0.003). Patients with anti‐Th/To antibodies had more subtle skin changes, less severe vascular involvement, and less frequent distal esophageal hypomotility. Both groups had a higher frequency of “intrinsic” pulmonary hypertension than has been previously reported in the literature (28% and 19% of anti‐Th/To–positive and ACA‐positive patients, respectively), perhaps due to referral bias. Patients in the anti‐Th/To group more often had radiographic evidence of interstitial lung disease (48% versus 13% of the ACA group; P < 0.0001). Scleroderma renal crisis was uncommon (4 cases), but occurred exclusively in the anti‐Th/To group. Survival among the anti‐Th/To–positive patients was reduced compared with that in the ACA group (P < 0.02).

Conclusion

Patients with anti‐Th/To and those with ACA most often develop lcSSc and have a high frequency of “intrinsic” pulmonary hypertension. Compared with the ACA patients, anti‐Th/To lcSSc patients have more subtle cutaneous, vascular, and gastrointestinal involvement, but more often have certain features typically seen in diffuse scleroderma, such as pulmonary fibrosis and scleroderma renal crisis, as well as reduced survival.
High‐cell‐density fed‐batch cultivation of the docosahexaenoic acid producing marine alga Crypthecodinium cohnii
Authors: De Swaaf Martin E. , Sijtsma Lolke , Pronk Jack T.
Published: January 2003

The heterotrophic marine alga Crypthecodinium cohnii is known to produce docosahexaenoic acid (DHA), a polyunsaturated fatty acid with food and pharmaceutical applications, during batch cultivation on complex media containing sea salt, yeast extract, and glucose. In the present study, fed‐batch cultivation was studied as an alternative fermentation strategy for DHA production. Glucose and acetic acid were compared as carbon sources. For both substrates, the feed rate was adapted to the maximum specific consumption rate of C. cohnii. In glucose‐grown cultures, this was done by maintaining a significant glucose concentration (between 5 and 20 g/L) throughout fermentation. In acetic acid‐grown cultures, the medium feed was automatically controlled via the culture pH. A feed consisting of acetic acid (50% w/w) resulted in a higher overall volumetric productivity of DHA (rDHA) than a feed consisting of 50% (w/v) glucose (38 and 14 mg/L/h, respectively). The rDHA was further increased to 48 mg/L/h using a feed consisting of pure acetic acid. The latter fermentation strategy resulted in final concentrations of 109 g/L dry biomass, 61 g/L lipid, and 19 g/L DHA. These are the highest biomass, lipid, and DHA concentrations reported to date for a heterotrophic alga. Vigorous mixing was required to sustain aerobic conditions during high‐cell‐density cultivation. This was complicated by culture viscosity, which resulted from the production of viscous extracellular polysaccharides. These may present a problem for large‐scale industrial production of DHA. Addition of a commercial polysaccharide‐hydrolase preparation could decrease the viscosity of the culture and the required stirring. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 666–672, 2003.
Potential inhibitors from wet oxidation of wheat straw and their effect on ethanol production of Saccharomyces cerevisiae: Wet oxidation and fermentation by yeast
Authors: Klinke H. B. , Olsson L. , Thomsen A. B. , Ahring B. K.
Published: January 2003

Alkaline wet oxidation (WO) (using water, 6.5 g/L sodium carbonate and 12 bar oxygen at 195°C) was used as pretreatment method for wheat straw (60 g/L), resulting in a hydrolysate and a cellulosic solid fraction. The hydrolysate consisted of soluble hemicellulose (8 g/L), low‐molecular‐weight carboxylic acids (3.9 g/L), phenols (0.27 g/L = 1.7 mM) and 2‐furoic acid (0.007 g/L). The wet oxidized wheat straw hydrolysate caused no inhibition of ethanol production by Saccharomyces cerevisiae ATCC 96581. Nine phenols and 2‐furoic acid, identified to be present in the hydrolysate, were each tested in concentrations of 50–100 times the concentration found in the hydrolysate for their effect on fermentation by yeast. At these high concentrations (10 mM), 4‐hydroxybenzaldehyde, vanillin, 4‐hydroxyacetophenone and acetovanillone caused a 53–67% decrease in the volumetric ethanol productivity in S. cerevisiae compared to controls with an ethanol productivity of 3.8 g/L. The phenol acids (4‐hydroxy, vanillic and syringic acid), 2‐furoic acid, syringaldehyde and acetosyringone were less inhibitory, causing a 5–16% decrease in ethanol productivity. By adding the same aromatic compounds to hydrolysate (10 mM), it was shown that syringaldehyde and acetovanillone interacted negatively with hydrolysate components on the ethanol productivity. Fermentation in WO hydrolysate, that had been concentrated 6 times by freeze‐drying, lasted 4 hours longer than in regular hydrolysate; however, the ethanol yield was the same. The longer fermentation time could not be explained by an inhibitory action of phenols alone, but was more likely caused by inhibitory interactions of phenols with carboxylic acids, such as acetic and formic acid. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 738–747, 2003.
Pre-emptive surgery and increasing demands for technical perfection
Authors: Brennan Murray F
Published: January 2003

Challenging times for surgeons
Current practice of emergency vagotomy and Helicobacter pylori eradication for complicated peptic ulcer in the United Kingdom
Authors: Gilliam A D , Speake W J , Lobo D N , Beckingham I J
Published: January 2003

Background

The aim was to assess the current opinion of surgeons, by subspecialty, towards vagotomy and the practice of Helicobacter pylori testing, treatment and follow-up, in patients with bleeding or perforated duodenal ulcer.

Methods

A postal questionnaire was sent to 1073 Fellows of the Association of Surgeons of Great Britain and Ireland in 2001.

Results

Some 697 valid questionnaires were analysed (65·0 per cent). Most surgeons did not perform vagotomy for perforated or bleeding duodenal ulcer. There was no statistical difference between the responses of upper gastrointestinal surgeons and those of other specialists for perforated (P = 0·35) and bleeding (P = 0·45) ulcers. Respondents were more likely to perform a vagotomy for bleeding than for a perforated ulcer (P < 0·001). Although more than 80 per cent of surgeons prescribed H. pylori eradication treatment after operation, fewer than 60 per cent routinely tested patients for H. pylori eradication. Upper gastrointestinal surgeons were more likely to prescribe H. pylori treatment and test for eradication than other specialists (P < 0·01).

Conclusion

Most surgeons in the UK no longer perform vagotomy for duodenal ulcer complications.