bluuebunny/crossref_metadata_2025_split huggingface dataset:
Newly developed microsatellite markers for the pan‐European duck mussel, Anodonta anatina: revisiting the main mitochondrial lineages
Authors: Lopes‐Lima Manuel , Sousa Ronaldo , Teixeira Amílcar , Varandas Simone , Riccardi Nicoletta , Aldridge David C. , Froufe Elsa 
Published: June 2015

Abstract

Freshwater mussels of the family Unionidae are one of the most threatened groups worldwide and have suffered severe decline over recent decades. Although the freshwater duck mussel, Anodonta anatina (Linnaeus, 1758), is still widespread, this species has shown evidence of recent declines and is already protected in some European countries.
Informed conservation efforts must take into account patterns in genetic diversity and phylogeography. In the present study, 20 newly developed polymorphic loci were described and tested in seven populations of A. anatina, belonging to three previously detected divergent mtDNA lineages. The genetic diversity patterns, within and among A. anatina populations, were evaluated to test their congruence with those lineages.
A high genetic differentiation (FST) was found among all populations, with the exception of those in Central Europe (Germany) and the UK, which were not strongly structured.
The present study confirms the division of the species into three evolutionarily significant units corresponding to the three previously detected mtDNA lineages, which should be managed independently. Furthermore, owing to the high differentiation among southern European populations, the establishment of distinct management units for the Guadalquivir, Guadiana and Douro populations in the Iberian Peninsula is also proposed.
Copyright © 2015 John Wiley & Sons, Ltd.
A BAFF Receptor His159Tyr Mutation in Sjögren's Syndrome–Related Lymphoproliferation
Authors: Papageorgiou Aristea , Mavragani Clio P. , Nezos Andrianos , Zintzaras Elias , Quartuccio Luca , De Vita Salvatore , Koutsilieris Michael , Tzioufas Athanasios G. , Moutsopoulos Haralampos M. , Voulgarelis Michael 
Published: June 2015

ObjectiveTo study the prevalence, clinical associations, and functional implications of the His159Tyr mutation of the BAFF receptor (BAFF‐R) in patients with Sjögren's syndrome (SS).MethodsThe BAFF‐R His159Tyr mutation was evaluated using polymerase chain reaction (PCR)–based assays in 247 patients with SS (of whom 70 had SS complicated by lymphoma [SS‐lymphoma]), 145 with systemic lupus erythematosus (SLE), and 101 with rheumatoid arthritis (RA), as well as 180 healthy controls. Real‐time PCR and Western blotting were performed for the quantification of both NF‐κB1 and NF‐κB2 messenger RNA (mRNA) transcript and protein levels in isolated B cells from patients with SS‐lymphoma carrying the mutation (SS‐lymphoma–BAFF‐RHis159Tyr–derived B cells) compared to B cells from patients with SS‐lymphoma who were not carriers of the mutation and healthy controls.ResultsBoth the SS‐lymphoma and SS‐nonlymphoma patient subgroups exhibited significantly higher frequencies of the His159Tyr BAFF‐R mutation compared to healthy controls (8.6% of SS‐lymphoma patients and 6.2% of SS‐nonlymphoma patients versus 1.7% of healthy controls; P = 0.02 and P = 0.04, respectively). The corresponding frequencies of the His159Tyr BAFF‐R mutation in SLE and RA patients were 3.5% and 3%, respectively. Of interest, 71.4% of the SS patients with mucosa‐associated lymphoid tissue (MALT) lymphoma who were between the ages of 31 and 40 years at disease onset were mutation carriers. The generalized odds ratio for the development of SS‐related MALT lymphoma in the younger age at onset (age <40 years) group in the presence of the BAFF‐R mutation was 6.1 (95% confidence interval 2.0–18.7) (P < 0.01). Expression of NF‐κB at both the mRNA and protein level was up‐regulated in SS‐lymphoma–BAFF‐RHis159Tyr–derived B cells.ConclusionThis study identifies an increased prevalence of the BAFF‐R His159Tyr mutation in patients with SS, particularly in those with SS complicated by MALT lymphoma whose disease onset occurred at a younger age. BAFF‐RHis159Tyr–mediated activation of the alternate NF‐κB pathway might contribute to the pathogenesis of SS‐related lymphoproliferative disease.
Rational Design of Antirheumatic Prodrugs Specific for Sites of Inflammation
Authors: Onuoha Shimobi C. , Ferrari Mathieu , Sblattero Daniele , Pitzalis Costantino 
Published: June 2015

ObjectiveBiologic drugs, such as the anti–tumor necrosis factor (anti‐TNF) antibody adalimumab, have represented a breakthrough in the treatment of rheumatoid arthritis. Yet, concerns remain over their lack of efficacy in a sizable proportion of patients and their potential for systemic side effects such as infection. Improved biologic prodrugs specifically targeted to the site of inflammation have the potential to alleviate current concerns surrounding biologic anticytokine therapies. The purpose of this study was to design, construct, and evaluate in vitro and ex vivo the targeting and antiinflammatory capacity of activatable bispecific antibodies.MethodsActivatable dual variable domain (aDVD) antibodies were designed and constructed to target intercellular adhesion molecule 1 (ICAM‐1), which is up‐regulated at sites of inflammation, and anti‐TNF antibodies (adalimumab and infliximab). These bispecific molecules included an external arm that targets ICAM‐1 and an internal arm that comprises the therapeutic domain of an anti‐TNF antibody. Both arms were linked to matrix metalloproteinase (MMP)–cleavable linkers. The constructs were tested for their ability to bind and neutralize both in vitro and ex vivo targets.ResultsIntact aDVD constructs demonstrated significantly reduced binding and anti‐TNF activity in the prodrug formulation as compared to the parent antibodies. Human synovial fluid and physiologic concentrations of MMP enzyme were capable of cleaving the external domain of the antibody, revealing a fully active molecule. Activated antibodies retained the same binding and anti‐TNF inhibitory capacities as the parent molecules.ConclusionThe design of a biologic prodrug with enhanced specificity for sites of inflammation (synovium) and reduced specificity for off‐target TNF is described. This construct has the potential to form a platform technology that is capable of enhancing the therapeutic index of drugs for the treatment of RA and other inflammatory diseases.
Interleukin‐21 Receptor Blockade Inhibits Secondary Humoral Responses and Halts the Progression of Preestablished Disease in the (NZB × NZW)F1 Systemic Lupus Erythematosus Model
Authors: Zhang Ming , Yu Gang , Chan Brian , Pearson Joshua T. , Rathanaswami Palaniswami , Delaney John , Ching Lim Ai , Babcook John , Hsu Hailing , Gavin Marc A. 
Published: June 2015

ObjectiveSystemic lupus erythematosus (SLE) is a complex autoimmune disease that is driven in part by chronic B and T lymphocyte hyperresponsiveness to self antigens. A deficiency of interleukin‐21 (IL‐21) or IL‐21 receptor (IL‐21R) in mice dramatically reduces inflammation and B and T cell activation in models of autoimmunity, including SLE. However, whether IL‐21 is essential for the maintenance and amplification of preestablished inflammation has not been widely examined in various animal models. The purpose of this study was to examine the impact of novel mouse IL‐21R neutralizing antibodies on recall responses to antigen challenge and on disease progression in the (NZB × NZW)F1 (NZB/NZW) mouse model of SLE.MethodsHumoral and cellular immune responses to immunization with sheep red blood cells (SRBCs) were measured in mice dosed with IL‐21R blocking antibodies. Progression of nephritis and markers of immune activation was monitored in NZB/NZW mice following different anti–IL‐21R treatment regimens.ResultsIL‐21R blockade specifically inhibited secondary IgG responses to SRBC immunization. In NZB/NZW mice, IL‐21R blockade completely inhibited the onset of nephritis, which was associated with dramatic reductions in splenomegaly and in B cell and T cell activation. When administered to mice with preexisting disease, anti–IL‐21R antibody halted the disease progression and mortality and reversed the nephritis in a subset of mice. Furthermore, treatment cessation was not followed by rapid reemergence of disease.ConclusionOur results highlight the importance of IL‐21 in promoting humoral recall responses and in sustaining autoimmune inflammation.
Long‐Term Safety, Efficacy, and Quality of Life in Patients With Juvenile Idiopathic Arthritis Treated With Intravenous Abatacept for Up to Seven Years
Authors: Lovell Daniel J. , Ruperto Nicolino , Mouy Richard , Paz Eliana , Rubio‐Pérez Nadina , Silva Clovis A. , Abud‐Mendoza Carlos , Burgos‐Vargas Ruben , Gerloni Valeria , Melo‐Gomes Jose A. , Saad‐Magalhaes Claudia , Chavez‐Corrales J. , Huemer Christian , Kivitz Alan , Blanco Francisco J. , Foeldvari Ivan , Hofer Michael , Huppertz Hans‐Iko , Job Deslandre Chantal , Minden Kirsten , Punaro Marilynn , Block Alan J. , Giannini Edward H. , Martini Alberto ,   for the Pediatric Rheumatology Collaborative Study Group and the Paediatric Rheumatology International Trials Organisation
Published: June 2015

ObjectiveThe efficacy and safety of abatacept in patients with juvenile idiopathic arthritis (JIA) who experienced an inadequate response to disease‐modifying antirheumatic drugs were previously established in a phase III study that included a 4‐month open‐label lead‐in period, a 6‐month double‐blind withdrawal period, and a long‐term extension (LTE) phase. The aim of this study was to present the safety, efficacy, and patient‐reported outcomes of abatacept treatment (10 mg/kg every 4 weeks) during the LTE phase, for up to 7 years of followup.MethodsPatients enrolled in the phase III trial could enter the open‐label LTE phase if they had not achieved a response to treatment at month 4 or if they had received abatacept or placebo during the double‐blind period.ResultsOne hundred fifty‐three (80.5%) of 190 patients entered the LTE phase, and 69 patients (36.3%) completed it. The overall incidence rate (events per 100 patient‐years) of adverse events decreased during the LTE phase (433.61 events during the short‐term phase [combined lead‐in and double‐blind periods] versus 132.39 events during the LTE phase). Similar results were observed for serious adverse events (6.82 versus 5.60), serious infections (1.13 versus 1.72), malignancies (1.12 versus 0), and autoimmune events (2.26 versus 1.18). American College of Rheumatology (ACR) Pediatric 30 (Pedi 30) responses, Pedi 70 responses, and clinically inactive disease status were maintained throughout the LTE phase in patients who continued to receive therapy. Improvements in the Child Health Questionnaire physical and psychosocial summary scores were maintained over time.ConclusionLong‐term abatacept treatment for up to 7 years was associated with consistent safety, sustained efficacy, and quality‐of‐life benefits in patients with JIA.
Use of Ultrasound‐Guided Small Joint Biopsy to Evaluate the Histopathologic Response to Rheumatoid Arthritis Therapy: Recommendations for Application to Clinical Trials
Authors: Humby Frances , Kelly Stephen , Hands Rebecca , Rocher Vidalba , DiCicco Maria , Ng Nora , Zou Lu , Bugatti Serena , Manzo Antonio , Caporali Roberto , Montecucco Carlomaurizio , Bombardieri Michele , Pitzalis Costantino 
Published: June 2015

ObjectiveTo examine in a cohort of rheumatoid arthritis (RA) patients undergoing serial ultrasound (US)–guided biopsies of small joints in the context of clinical trials whether sufficient synovial tissue could be obtained at both baseline and second biopsy to: 1) accurately evaluate the synovial immune phenotype, 2) permit adequate RNA extraction to determine molecular signatures, and 3) sensitively detect change in the number of synovial sublining macrophages (CD68+) following effective therapy.MethodsSynovial samples from RA patients undergoing US‐guided biopsy of small joints as part of 2 clinical trials (Barts Early Arthritis Cohort [n = 18] and the Clinical and Pathological Response to Certolizumab Pegol (CLIP‐Cert) study [n = 17]) were examined, and the quality and quantity of histologic samples and RNA extracted per joint were determined and compared to synovial thickness and power Doppler scores determined by US before biopsy. Modulation of the number of CD68+ sublining macrophages was correlated with clinical response to treatment.ResultsGood quality synovial tissue that accurately reflected the synovial immune phenotype of the total joint was obtained in 80% of US‐guided procedures when synovial thickness (higher than grade 2) was documented before biopsy. In 100% of the procedures, sufficient RNA was extracted to permit molecular analysis. There was a significant correlation between change in CD68+ sublining macrophage number and clinical response to treatment.ConclusionThis study provides minimum standards for sample retrieval for small joint biopsy. Furthermore, our findings confirm the clinical utility of the procedure in the largest reported cohort of US‐guided small joint biopsies. The demonstration that small joint synovial tissue can be readily accessed by a technically simple, minimally invasive procedure is likely to facilitate critical advancements in the knowledge of RA pathobiology and personalized health care.
Promotion of Hypercoagulability in Antineutrophil Cytoplasmic Antibody–Associated Vasculitis by C5a‐Induced Tissue Factor–Expressing Microparticles and Neutrophil Extracellular Traps
Authors: Huang Yi‐Min , Wang Huan , Wang Chen , Chen Min , Zhao Ming‐Hui 
Published: June 2015

ObjectivePatients with antineutrophil cytoplasmic antibody–associated vasculitis (AAV) have a high frequency of venous thromboembolic events and a hypercoagulable state. As C5a‐primed neutrophils play an important role in the development of AAV, we investigated whether C5a‐induced neutrophil tissue factor (TF)–expressing microparticles (MPs) and neutrophil extracellular traps (NETs) might promote hypercoagulability in AAV.MethodsTF‐expressing MPs were measured by flow cytometry. TF‐expressing NETs were assessed by confocal microscopy. Levels of thrombin–antithrombin complexes were determined by enzyme‐linked immunosorbent assay. The effect of C5a in sera from AAV patients was evaluated by treating neutrophils with C5a receptor antagonist before incubation with sera from AAV patients with active disease.ResultsTreatment of C5a‐primed neutrophils with antineutrophil cytoplasmic antibody (ANCA)–positive IgG resulted in the release of TF‐bearing MPs and NETs. Neutrophils from healthy donors treated with sera from patients with active AAV released TF‐bearing MPs and NETs, which were abolished by treatment with C5a receptor antagonist. Involvement of TF in MP‐ or NET‐dependent thrombin generation was indicated by the findings of antibody neutralization studies. NETs with thrombin‐generating capacity were demonstrated by DNase I treatment.ConclusionC5a‐primed neutrophils produce TF‐expressing MPs and NETs after stimulation with ANCAs, indicating a mechanism for hypercoagulability in AAV that was not previously recognized.
Brief Report: Association of CCR1, KLRC4, IL12A–AS1, STAT4, and ERAP1 With Behçet's Disease in Iranians
Authors: Sousa Inês , Shahram Farhad , Francisco David , Davatchi Fereydoun , Abdollahi Bahar Sadeghi , Ghaderibarmi Fahmida , Nadji Abdolhadi , Mojarad Shafiee Niloofar , Xavier Joana M. , Oliveira Sofia A. 
Published: June 2015

ObjectiveTo independently replicate the top findings from 4 published genome‐wide association studies (GWAS) of susceptibility genes in Behçet's disease (BD).MethodsWe tested 14 single‐nucleotide polymorphisms (SNPs) in 13 genomic loci (excluding the major histocompatibility complex [MHC], IL10, and IL23R–IL12RB2, which have already been associated with BD in Iranians) for allelic and genotypic associations with BD in 973 patients and 828 controls from Iran and performed meta‐analyses of the significantly associated markers.ResultsSix SNPs (in decreasing order of significance, rs7616215 located 38 kb downstream of CCR1, rs2617170 [p.Asn104Ser] in KLRC4, rs17810546 in IL12A–AS1, rs7574070 in STAT4, and rs10050860 [p.Asp575Asn] and rs13154629 in ERAP1) were nominally associated with BD in both allelic association tests (5.05 × 10−9 ≤ Pallele ≤ 7.55 × 10−3) and sex‐adjusted genotypic association tests (6.01 × 10−9 ≤ adjusted P value ≤ 1.30 × 10−2). For all 6 SNPs tested by meta‐analysis (Pmeta), the association with BD was strengthened, because the direction and magnitude of association were similar across populations (e.g., for rs7574070, odds ratio [OR] for A allele 1.29 [95% confidence interval (95% CI) 1.21–1.37], Pmeta = 2.34 × 10−16; for rs7616215, OR for C allele 0.70 [95% CI 0.65–0.76], Pmeta = 1.54 × 10−19; for rs17810546, OR for A allele 0.60 [95% CI 0.52–0.70], Pmeta = 6.34 × 10−11; for rs2617170, OR for T allele 0.76 [95% CI 0.70–0.81], Pmeta = 2.75 × 10−14; for rs13154629, OR for TT genotype 2.76 [95% CI 2.01–3.80], Pmeta = 3.57 × 10−10).ConclusionThis study reinforces the notion that CCR1, KLRC4, IL12A–AS1, STAT4, and ERAP1 are bona fide susceptibility genes for BD, in addition to the MHC, IL10, and IL23R–IL12RB2 loci. Future genetic and functional studies are now warranted to uncover the roles of these genes in the pathogenesis of BD.
Increased Expression of NAPDH Oxidase 4 in Systemic Sclerosis Dermal Fibroblasts: Regulation by Transforming Growth Factor β
Authors: Piera‐Velazquez Sonsoles , Makul Alma , Jiménez Sergio A. 
Published: June 2015

ObjectiveSystemic sclerosis (SSc) is characterized by severe and often progressive fibrosis of the skin and multiple internal organs. The mechanisms responsible for these alterations remain obscure, although excessive reactive oxygen species (ROS)–mediated oxidative stress has been implicated. NOX‐4 is 1 of 7 isoforms of NADPH oxidase responsible for the generation of ROS. The purpose of this study was to examine NOX‐4 expression in skin and cultured dermal fibroblasts from SSc patients and to examine its regulation by transforming growth factor β1 (TGFβ1).MethodsNOX‐4 was assessed in normal and SSc skin by immunohistologic analysis and in normal and SSc cultured dermal fibroblasts by quantitative polymerase chain reaction analysis, fluorescence microscopy, and Western blotting. ROS levels were assessed by fluorescence measurement of H2O2 production. Specific kinase inhibitors were used to study the TGFβ1 signaling involved in NOX‐4 stimulation. NOX‐4 inhibition/down‐regulation was induced with a selective NOX‐4 small‐molecule inhibitor and NOX‐4 small interfering RNA (siRNA).ResultsIn contrast with normal skin fibroblasts, those from SSc skin showed intense NOX‐4 staining. Cultured SSc fibroblasts displayed increased NOX‐4 expression. TGFβ1 caused potent NOX‐4 protein and messenger RNA stimulation in normal and SSc fibroblasts, which was mediated by the protein kinase Cδ (PKCδ) and Smad2/3 pathways. NOX‐4 knockdown in SSc fibroblasts reduced the production of ROS and lowered the expression of type I collagen.ConclusionNOX‐4 expression and production were found to be constitutively elevated in SSc skin and cultured SSc dermal fibroblasts. TGFβ1 stimulated NOX‐4 expression in normal and SSc fibroblasts through PKCδ and Smad2/3 signaling pathways. A small‐molecule NOX‐4 inhibitor decreased collagen and fibronectin production by normal and SSc fibroblasts, and NOX‐4 siRNA knockdown reduced ROS and collagen production by SSc fibroblasts. These results demonstrate the involvement of NOX‐4 in SSc‐associated fibrosis and indicate NOX‐4 inhibitors as novel therapeutic approaches for SSc.
Articular Cartilage Endurance and Resistance to Osteoarthritic Changes Require Transcription Factor Erg
Authors: Ohta Yoichi , Okabe Takahiro , Larmour Colleen , Di Rocco Agnese , Maijenburg Marijke W. , Phillips Amanda , Speck Nancy A. , Wakitani Shigeyuki , Nakamura Takashi , Yamada Yoshihiko , Enomoto‐Iwamoto Motomi , Pacifici Maurizio , Iwamoto Masahiro 
Published: June 2015

ObjectiveTo determine whether and how the transcription factor Erg participates in the genesis, establishment, and maintenance of articular cartilage.MethodsFloxed Erg mice were mated with Gdf5‐Cre mice to generate conditional mutants lacking Erg in their joints. Joints of mutant and control mice were subjected to morphologic and molecular characterization and also to experimental surgically induced osteoarthritis (OA). Gene expression, promoter reporter assays, and gain‐ and loss‐of‐function in vitro tests were used to characterize molecular mechanisms of Erg action.ResultsConditional Erg ablation did not elicit obvious changes in limb joint development and overall phenotype in juvenile mice. However, as mice aged, joints of mutant mice degenerated spontaneously and exhibited clear OA‐like phenotypic defects. Joints in juvenile mutant mice were more sensitive to surgically induced OA and became defective sooner than operated joints in control mice. Global gene expression data and other studies identified parathyroid hormone–related protein (PTHrP) and lubricin as possible downstream effectors and mediators of Erg action in articular chondrocytes. Reporter assays using control and mutated promoter‐enhancer constructs indicated that Erg acted on Ets DNA binding sites to stimulate PTHrP expression. Erg was up‐regulated in severely affected areas in human OA articular cartilage but remained barely appreciable in areas of less affected cartilage.ConclusionThe study shows for the first time that Erg is a critical molecular regulator of the endurance of articular cartilage during postnatal life and that Erg can mitigate spontaneous and experimental OA. Erg appears to do this through regulating expression of PTHrP and lubricin, factors known for their protective roles in joints.
Dysregulated Osteoclastogenesis Is Related to Natural Killer T Cell Dysfunction in Rheumatoid Arthritis
Authors: Jin Hye‐Mi , Kee Seung‐Jung , Cho Young‐Nan , Kang Jeong‐Hwa , Kim Moon‐Ju , Jung Hyun‐Ju , Park Ki‐Jeong , Kim Tae‐Jong , Lee Sang‐Il , Choi Hyuck , Koh Jeong‐Tae , Kim Nacksung , Park Yong‐Wook 
Published: June 2015

ObjectiveTo investigate the role played by natural killer T (NKT) cells in osteoclastogenesis and their effects on inflammatory bone destruction.MethodsPatients with rheumatoid arthritis (RA) (n = 25) and healthy controls (n = 12) were enrolled in this study. In vitro osteoclastogenesis experiments were performed using peripheral blood mononuclear cells (PBMCs) in the presence of macrophage colony‐stimulating factor and RANKL. PBMCs were cultured in vitro with α‐galactosylceramide (αGalCer), and proliferation indices of NKT cells were estimated by flow cytometry. In vivo effects of αGalCer‐stimulated NKT cells on inflammation and bone destruction were determined in mice with collagen‐induced arthritis.ResultsIn vitro osteoclastogenesis was found to be significantly inhibited by αGalCer in healthy controls but not in RA patients. Proliferative responses of NKT cells and STAT‐1 phosphorylation in monocytes in response to αGalCer were impaired in RA patients. Notably, αGalCer‐stimulated NKT cells inhibited osteoclastogenesis mainly via interferon‐γ production in a cytokine‐dependent manner (not by cell–cell contact) and down‐regulated osteoclast‐associated genes. Mice treated with αGalCer showed less severe arthritis and reduced bone destruction. Moreover, proinflammatory cytokine expression in arthritic joints was found to be reduced by αGalCer treatment.ConclusionThis study primarily demonstrates that αGalCer‐stimulated NKT cells have a regulatory effect on osteoclastogenesis and a protective effect against inflammatory bone destruction. However, it also shows that these effects of αGalCer are diminished in RA patients and that this is related to NKT cell dysfunction. These findings provide important information for those searching for novel therapeutic strategies to prevent bone destruction in RA.