bluuebunny/crossref_metadata_2025_split huggingface dataset:
Functional characterization of adherent synovial fluid cells in rheumatoid arthritis: Destructive potential in vitro and in vivo
Authors: Neidhart Michel , Seemayer Christian A. , Hummel Klaus M. , Michel Beat A. , Gay Renate E. , Gay Steffen 
Published: July 2003

AbstractObjectiveTo characterize the morphologic and immunologic features of adherent synovial fluid cells derived from patients with rheumatoid arthritis (RA), and to explore their potential function in vitro and in vivo by focusing on cartilage destruction.MethodsSynovial fluid adherent cells obtained from patients with RA and from control subjects were characterized by immunohistochemistry, flow cytometry, and electron microscopy. In vitro, these cells were cultured in the presence of cartilage particles. Cartilage destruction was monitored by the release of sulfated glycosaminoglycans (sGAG) into the medium, and the level of matrix metalloproteinase 1 (MMP‐1) in the cell culture supernatant was measured by enzyme‐linked immunosorbent assay. To inhibit cartilage destruction in vitro, the MMP inhibitor marimastat was tested in this system. In vivo, in the SCID mouse coimplantation model, RA synovial fluid adherent cells and RA synovial fibroblasts (as positive controls) were coimplanted with human cartilage under the kidney capsule and maintained there for 60 days.ResultsIn vitro, the synovial fluid adherent cells consisted of 2 subpopulations, large round‐shaped macrophage‐like cells (CD68+) and spindle‐shaped fibroblast‐like cells (Thy‐1+). When passaged, the latter cells proliferated and organized themselves into 3‐dimensional formations. This allowed them to reach collagen particles fixed with agarose. Fibroblasts derived from synovial tissues could not be used in this assay because they grew only in monolayers and not on agarose. The majority (>90%) of passaged RA synovial fluid adherent cells expressed the Thy‐1+,CD45−,CD68−,CD86− phenotype. Electron microscopy did not reveal important morphologic differences between the 2 types of fibroblasts, those from synovial tissue or those from synovial fluid. However, synovial fluid adherent cells expressed lower levels of adhesion molecules, including CD54 and galectin 3, as well as the complement‐regulatory molecule CD55. The in vitro release of sGAG associated with cell activity was 2.5‐fold higher from RA synovial fluid adherent cells in comparison with that from negative control cells. The release of sGAG correlated with the concentration of MMP‐1 and was inhibited by the broad‐range MMP inhibitor marimastat in a dose‐dependent manner. RA synovial fluid adherent cells coimplanted with cartilage in SCID mice showed the same invasive behavior as that displayed by tissue‐derived RA synovial fibroblasts.ConclusionSimilar to tissue‐derived RA synovial fibroblasts, RA synovial fluid adherent cells, which contain “floating” anchorage‐independent fibroblast‐like cells, mediate cartilage destruction independent of the hyperplastic synovial tissue.
Polymorphism at position −308 of the tumor necrosis factor α gene influences outcome of infliximab therapy in rheumatoid arthritis
Authors: Mugnier Benedicte , Balandraud Nathalie , Darque Albert , Roudier Chantal , Roudier Jean , Reviron Denis 
Published: July 2003

AbstractObjectiveTo test whether the G‐to‐A polymorphism at position −308 in the promoter of the tumor necrosis factor α (TNFα) gene influences response to infliximab therapy in patients with rheumatoid arthritis (RA).MethodsWe genotyped 59 RA patients by polymerase chain reaction and subdivided them into two groups: those with the A/A or A/G genotype and those with the G/G genotype. We compared the groups' clinical responses to infliximab treatment after 22 weeks, using the Disease Activity Score in 28 joints (DAS28).ResultsWe found that 42% of patients in the A/A and A/G group and 81% of patients in the G/G group had improvement of at least 1.2 in the DAS28 score (P = 0.0086). The average improvement in the DAS28 score was 1.24 in the A/A and A/G patients and 2.29 in the G/G patients (P = 0.029).ConclusionThese data suggest that patients with a TNFα −308G/G genotype are better infliximab responders than are patients with A/A or A/G genotypes. TNFα −308 genotyping may be a useful tool for predicting response to infliximab treatment.
Five‐year followup of rheumatoid arthritis patients after early treatment with disease‐modifying antirheumatic drugs versus treatment according to the pyramid approach in the first year
Authors: Verstappen Suzan M. M. , Jacobs Johannes W. G. , Bijlsma Johannes W. J. , Heurkens Anton H. M. , van Booma‐Frankfort Christina , Borg Evert Jan ter , Hofman Dick M. , van der Veen Maaike J. 
Published: July 2003

AbstractObjectiveTo evaluate whether the clinical advantages observed after 1 year in a randomized controlled clinical trial, in which 2 treatment strategies were compared (the early disease‐modifying antirheumatic drug [DMARD] approach versus the pyramid approach), persist after 5 years.MethodsIn this study, 238 patients with recently diagnosed rheumatoid arthritis (RA) were randomized to either the pyramid group (n = 56) or the early DMARD group (n = 182). Patients assigned to the pyramid group received nonsteroidal antiinflammatory drugs for at least 1 year after inclusion (the mean ± SD lag time until first prescription of a DMARD was 14 ± 9 months). Patients in the early DMARD group were treated with a DMARD immediately after inclusion.ResultsAfter 5 years, data were available for 44 patients in the pyramid group (79%) and 145 patients in the early DMARD group (80%). No prolongation of the clinical advantages in favor of the early DMARD group, as observed after the first year, was demonstrated. Nevertheless, a significantly shorter delay time until complete response and a higher number of patients with overall clinically relevant improvement at several assessment points were observed in the early DMARD group compared with the pyramid group.ConclusionThe clinical results in favor of the early DMARD group, as observed after the first year, were not as evident after 5 years. This indicates that a more aggressive treatment approach in early RA is required, and that treatment should be continued for a prolonged period of time, in order to maintain the advantages obtained in the first year.
Expression of activation‐induced, T cell–derived, and chemokine‐related cytokine/lymphotactin and its functional role in rheumatoid arthritis
Authors: Blaschke Sabine , Middel Peter , Dorner Brigitte G. , Blaschke Volker , Hummel Klaus M. , Kroczek Richard A. , Reich Kristian , Benoehr Peter , Koziolek Michael , Müller Gerhard A. 
Published: July 2003

AbstractObjectiveTo evaluate the possible role of activation‐induced, T cell–derived, and chemokine‐related cytokine (ATAC)/lymphotactin (Lptn) in the pathogenesis of rheumatoid arthritis (RA).MethodsATAC/Lptn levels in serum and synovial fluid samples were measured by sandwich enzyme‐linked immunosorbent assay. Expression of messenger RNA for ATAC/Lptn in synovial tissues was analyzed by reverse transcription–polymerase chain reaction (PCR) and by in situ hybridization, and was quantitated by real‐time PCR. The phenotype of peripheral blood mononuclear cells (PBMCs) expressing ATAC/Lptn was analyzed by intracellular cytokine staining and flow cytometry.ResultsLevels of ATAC/Lptn were similar in sera and synovial fluids from RA patients (n = 20) and osteoarthritis controls (n = 15). In phorbol myristate acetate/ionomycin–stimulated PBMCs, ATAC/Lptn expression was detected in CD8+ T cells and in a significantly increased proportion of CD4+,CD28− T cells from RA patients as compared with healthy controls. In synovial tissues, ATAC/Lptn was predominantly localized in CD3+ T cells in the sublining layer. Lymphocytes, synovial macrophages, and, unexpectedly, fibroblast‐like synoviocytes (FLS) were identified as major target cells for ATAC/Lptn in RA synovium, as determined by analysis of the ATAC/Lptn receptor XCR1. In vitro, ATAC/Lptn stimulation of FLS resulted in a marked down‐regulation of matrix metalloproteinase 2 production.ConclusionThese data indicate that in RA synovium, ATAC/Lptn is mainly produced by T cells. Considering its function as a lymphocyte‐specific chemoattractant, ATAC/Lptn might be a key modulator for T cell trafficking in the pathogenesis of RA. In addition, functional studies suggest that ATAC/Lptn may exert additional immunomodulatory effects in RA.
Mechanical effects of the intraarticular administration of high molecular weight hyaluronic acid plus phospholipid on synovial joint lubrication and prevention of articular cartilage degeneration in experimental osteoarthritis
Authors: Kawano Tsutomu , Miura Hiromasa , Mawatari Taro , Moro‐Oka Takaaki , Nakanishi Yoshitaka , Higaki Hidehiko , Iwamoto Yukihide 
Published: July 2003

AbstractObjectiveTo examine in vivo the effects of a mixture of high molecular weight hyaluronic acid (HA) plus phospholipids on joint lubrication and articular cartilage degeneration.MethodsExperimental osteoarthritis (OA) of the right knee was induced by anterior cruciate and medial collateral ligament transection in 40 rabbits. The animals were subjected to 8 consecutive weekly intraarticular administrations of high molecular weight HA (the HA200 group), conventional molecular weight HA (the HA80 group), or high molecular weight HA plus L‐δ dipalmitoyl phosphatidylcholine liposomes (the PHA group) and were killed 1 week after the final injection. The remaining transected right knees (the OA group) and randomly selected nontransected contralateral left knees (the control group) were collected simultaneously. Each group (n = 10) was divided into 2 equal subgroups, one of which was evaluated histologically while the other was subjected to a lubricating ability test using a pendulum friction tester.ResultsThe injected knees had a tendency to demonstrate less damage to the articular cartilage compared with the OA group, and the histologic findings in all groups except for the PHA group differed significantly from the control group. There was a significant difference in the mean ± SD friction coefficient between the control group (0.0100 ± 0.00300) and the OA (0.0206 ± 0.00649), HA200 (0.0190 ± 0.00427), and HA80 (0.0177 ± 0.00712) groups (P < 0.05 for each comparison), but not between the control group and the PHA group (0.0150 ± 0.00330) (P = 0.15).ConclusionTo our knowledge, this is the first in vivo study to examine whether intraarticular injections of phospholipids influence joint lubrication by acting as a boundary lubricant, thus protecting articular cartilage from degenerative changes.
Analysis of systemic sclerosis in twins reveals low concordance for disease and high concordance for the presence of antinuclear antibodies
Authors: Feghali‐Bostwick Carol , Medsger Thomas A. , Wright Timothy M. 
Published: July 2003

AbstractObjectiveTo examine concordance for systemic sclerosis (SSc) in monozygotic (MZ) and dizygotic (DZ) twins.MethodsMZ and DZ twins were recruited nationwide. Zygosity was confirmed by DNA fingerprint analysis. The presence of antinuclear antibodies (ANAs) was determined using indirect immunofluorescence with HEp‐2 cells as substrate. Identification of SSc‐associated serum autoantibodies was performed by immunoprecipitation and double immunodiffusion. Major histocompatibility complex class II alleles were identified by polymerase chain reaction–restriction fragment length polymorphism analysis.ResultsConcordance for SSc was found to be similar in MZ and DZ twins. Overall concordance for SSc was low in the twins (4.7%). Concordance for the presence of ANAs was significantly higher in MZ twins compared with DZ twins. SSc‐associated serum autoantibodies occurred exclusively in patients with SSc. The distribution of SSc‐associated serum autoantibodies was similar to that observed in our large database of SSc patients. Increased HLA allele sharing was detected in DZ twins, irrespective of disease concordance.ConclusionThese results indicate that inherited genetic factors are not sufficient to explain the development of SSc. Rather, these data indicate that inheritance may play a role in the development of serum autoantibodies in the “healthy” twin sibling of an SSc patient.
The spontaneous remission of juvenile idiopathic arthritis is characterized by CD30+ T cells directed to human heat‐shock protein 60 capable of producing the regulatory cytokine interleukin‐10
Authors: de Kleer I. M. , Kamphuis S. M. , Rijkers G. T. , Scholtens L. , Gordon G. , de Jager W. , Häfner R. , van de Zee R. , van Eden W. , Kuis W. , Prakken B. J. 
Published: July 2003

AbstractObjectiveTo test the hypothesis that T cell reactivity to self heat‐shock protein 60 (Hsp60) in patients with remitting juvenile idiopathic arthritis (JIA) is part of an antiinflammatory, regulatory mechanism.MethodsUsing peripheral blood–derived mononuclear cells (PBMCs) and synovial fluid–derived mononuclear cells (SFMCs) obtained from patients with JIA, we analyzed the expression of CD30 and the induction of regulatory cytokines in response to human and mycobacterial Hsp60.ResultsIn oligoarticular JIA patients, in vitro activation of PBMCs and SFMCs with Hsp60 induced a high expression of CD30 on CD4+, activated (HLA–DR–positive), memory (CD45RO+) T cells. The expression of CD30 induced by human Hsp60 was much higher than that induced by mycobacterial Hsp60. In oligoarticular JIA patients with active disease, the expression of CD30 in response to human Hsp60 was paralleled by a high interleukin‐10 (IL‐10):interferon‐γ (IFNγ) ratio. In addition, restimulated human Hsp60–specific T cell lines from oligoarticular JIA patients showed a high production of IL‐10 and a low production of IFNγ. In contrast, PBMCs and SFMCs from polyarticular JIA patients responded to human Hsp60 with virtually no expression of CD30 and a low IL‐10:IFNγ ratio.ConclusionThe results show that T cells responding to human Hsp60 in oligoarticular JIA patients express CD30, and during active phases of the disease, these T cells have a cytokine profile with a high IL‐10:IFNγ ratio. These findings suggest that in oligoarticular JIA patients, human Hsp60–specific CD4+ cells have a regulatory function and contribute to disease remission.
Clinical and physiological findings, and possible aetiological factors of rectal hyposensitivity
Authors: Gladman M A , Scott S M , Williams N S , Lunniss P J 
Published: July 2003

Abstract

Background
Rectal hyposensitivity (RH) relates to insensitivity of the rectum on anorectal physiological investigation and appears common in functional bowel disorders. The clinical significance of this physiological abnormality is unclear.

Method
RH was defined as one or more sensory threshold volumes raised beyond the normal range (mean plus two standard deviations) on rectal balloon distension. Clinical information and results of other anorectal physiological investigations were evaluated in 261 patients with RH.

Results
Patients with RH most commonly presented with constipation (48 per cent), constipation and incontinence in combination (27 per cent), or faecal incontinence (20 per cent). Thirty-eight per cent of patients had a history of previous pelvic surgery, 22 per cent a history of anal surgery and 13 per cent a history of spinal trauma. In patients with RH presenting with symptoms of constipation or incontinence, impaired rectal sensation was the only abnormality on physiological investigation in 48 per cent and 31 per cent respectively.

Conclusion
Patients with RH display marked heterogeneity in terms of presenting symptoms. The exact causes of RH are unknown, but there is evidence to suggest that pelvic nerve injury and spinal trauma are possible aetiological factors. RH appears important in the aetiology of both constipation and faecal incontinence, and may be useful as a predictor of surgical outcome.
Platinum(II)‐Based Coordination Compounds as Nucleic Acid Labeling Reagents: Synthesis, Reactivity, and Applications in Hybridization Assays
Authors: Heetebrij R. J. , Talman E. G. , Velzen M. A. v. , Gijlswijk R. P. M. v. , Snoeijers S. S. , Schalk M. , Wiegant J. , Rijke F. v.d. , Kerkhoven R. M. , Raap A. K. , Tanke H. J. , Reedijk J. , Houthoff H.‐J. 
Published: July 2003

AbstractThe synthesis, characterization, and molecular interactions of platinum(II) coordination compounds, which contain a distal nonradioactive reporter molecule, with mono‐ and polynucleotides are described. A [Pt(II)(en)(NH2(CH2)6NH‐tBoc)Cl](NO3) (en=ethylenediamine) entity has been coupled, after removal of the tBoc group, to a number of hapten and fluorophore molecules through succinimide derivatives. The influence of the various tethered reporter groups within these complexes on the reactivity towards guanosine 5′‐monophosphate (5′‐GMP), as a model for polynucleotide sequences, was investigated to shed light on the use of these reagents in hybridization assays. Reactivity turned out to be strongly dictated by the chemical nature of the distal reporter molecule present. At pH 7.0 the sequence of reactivity is cationic ≈ aromatic (stacking) > neutral > anionic; there is approximately an order of magnitude difference between the fastest reacting complex (k=10.2×10−2 M−1 s−1) and the slowest reacting complex (k=0.93×10−2 M−1 s−1) under these conditions. Platination of an oligodeoxynucleotide (30‐mer), dsDNA, or an RNA transcript, shows that a Pt/nucleotide ratio between 1:10 and 1:20 (established by using flameless atomic absorption spectroscopy) results in probes with excellent hybridization characteristics. In terms of applicability and detection limits these platinated nucleic acid probes perform equally well compared to conventionally generated nucleic acid probes, that is, through enzymatic incorporation of covalently labeled nucleotide triphosphates. Applications of these reagents to in situ hybridization assays and gene expression profiling on microarrays illustrate the potential of these monofunctional binding platinum triamine compounds.
Modified DNA Bearing 5(Methoxycarbonylmethyl)‐2′‐deoxyuridine: Preparation by PCR with Thermophilic DNA Polymerase and Postsynthetic Derivatization
Authors: Mehedi Masud Mohammad , Ozaki‐Nakamura Akiko , Kuwahara Masayasu , Ozaki Hiroaki , Sawai Hiroaki 
Published: July 2003

AbstractA thymidine analogue bearing a methyl ester at the C5 position was accepted as a substrate by the thermophilic family B DNA polymerases, KOD Dash, Pwo, and Vent(exo−), to form the corresponding PCR product, but not by the thermophilic family A DNA polymerases, Taq, Tth, and T7 thermosequenase. Modified DNA containing this analogue was prepared by PCR on a large scale with KOD Dash DNA polymerase and 5(methoxycarbonylmethyl)‐2′‐deoxyuridine 5′‐triphosphate as a substrate. The methyl ester of the modified DNA was further allowed to react with tris(2‐aminoethyl)amine or histamine by an ester–amide exchange reaction to form the corresponding derivatized DNA bearing a tris(2‐aminoethyl)amine or histamine moiety. Hydrolysis of the methyl ester of the modified DNA gave a functionalized DNA bearing an anionic carboxyl group. The derivatized DNA could act as a template for the PCR with KOD Dash DNA polymerase and the natural 2′‐deoxythymidine 5′‐triphosphate or the modified thymidine analogue as a substrate. The postsynthetic derivatization of the modified DNA may expand the variety of structurally modified DNA produced by PCR.
A Homology Model of Penicillin Acylase from Alcaligenes faecalis and In Silico Evaluation of its Selectivity
Authors: Braiuca Paolo , Ebert Cynthia , Fischer Lutz , Gardossi Lucia , Linda Paolo 
Published: July 2003

AbstractA three‐dimensional model of the relatively unknown penicillin acylase from Alcaligenes faecalis (PA‐AF) was built up by means of homology modeling based on three different crystal structures of penicillin acylase from various sources. An in silico selectivity study was performed to compare this homology model to the structure of the Escherichia coli enzyme (PA‐EC) in order to find any selectivity differences between the two enzymes. The program GRID was applied in combination with the principal component analysis technique to identify the regions of the active sites where the PAs potentially engage different interactions with ligands. These differences were further analyzed and confirmed by molecular docking simulations. The PA‐AF homology model provided the structural basis for the explanation of the different enantioselectivities of the enzymes previously demonstrated experimentally and reported in the literature. Different substrate selectivities were also predicted for PA‐AF compared to PA‐EC. Since no crystallographic data are available for PA‐AF to date, the three‐dimensional homology model represents a useful and efficient tool for fully exploiting this attractive and efficient biocatalyst, particularly in enantioselective acylations of amines.
Real‐time Detection of Nucleotide Incorporation During Complementary DNA Strand Synthesis
Authors: Krieg Alexander , Laib Stephan , Ruckstuhl Thomas , Seeger Stefan 
Published: July 2003

AbstractReal‐time observation of DNA strand synthesis by using a supercritical angle fluorescence detection apparatus for surface‐selective fluorescence detection is described. DNA template molecules were immobilized on a glass surface and the synthesis of the complementary strand was observed after addition of enzyme, dTTP, dATP, dGTP, and fluorescently labeled dCTP (d, deoxy; TP, triphosphate; T, A, G, and C, nucleobases). The fluorescence increase during the Klenow‐fragment‐catalyzed polymerization depends on the number of labeled dCTP nucleotides incorporated. The efficiency of this reaction is of the same order of magnitude as that of a bimolecular hybridization reaction.