The Minor Allele of rs7574865 in the STAT4 Gene Is Associated with Increased mRNA and Protein Expression

We studied 201 patients (80% female; median age, 54 years; median disease duration, 5.4 months) from PEARL study. Demographic, clinical, laboratory and therapeutic data were collected at each visit. IL-6 serum levels were measured by enzyme immune assay. The rs7574865 was genotyped using TaqMan probes. The expression levels of STAT4 mRNA were determined at 182 visits from 69 patients using quantitative real-time polymerase chain reaction. STAT4 protein was assessed by western blot in 62 samples from 34 patients. To determine the effect of different variables on the expression of STAT4 mRNA and protein, we performed multivariate longitudinal analyses using generalized linear models.This work was supported by grants awarded to IGA from the RETICS Program (RD08/0075/0004 and RD12/0009/0017 [RIER]) and FIS Program (PI11/0551) and to JM from RETICS Program (RD08/0075/0011 and RD12/0009/0004 [RIER]) from the Instituto de Salud Carlos III (www.isciii.es). Measurement of IL-6 levels described in this article was supported by different research grants from Roche to IGA. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer reviewe

VIP and CRF reduce ADAMTS expression and function in osteoarthritis synovial fibroblasts

ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family is known to play an important role in the pathogenesis of osteoarthritis (OA), working on aggrecan degradation or altering the integrity of extracellular matrix (ECM). Thus, the main purpose of our study was to define the role of vasoactive intestinal peptide (VIP) and corticotrophin-releasing factor (CRF), as immunoregulatory neuropeptides, on ADAMTS production in synovial fibroblasts (SF) from OA patients and healthy donors (HD). OA- and HD-SF were stimulated with pro-inflammatory mediators and treated with VIP or CRF. Both neuropeptides decreased ADAMTS-4, -5, -7 and -12 expressions, aggrecanase activity,glycosaminoglycans (GAG), and cartilage oligomeric matrix protein (COMP) degradation after stimulation with fibronectin fragments (Fn-fs) in OA-SF. After stimulation with interleukin-1b, VIP reduced ADAMTS-4 and -5, and both neuropeptides decreased ADAMTS-7 production and COMP degradation. Moreover, VIP and CRF reduced Runx2 and b-catenin activation in OA-SF. Our data suggest that the role of VIP and CRF on ADAMTS expression and cartilage degradation could be related to the OA pathology since scarce effects were produced in HD-SF. In addition,their effects might be greater when a degradation loop has been established, given that they were higher after stimulation with Fn-fs. Our results point to novel OA therapies based on the use of neuropeptides, since VIP and CRF are able to stop the first critical step, the loss of cartilageaggrecan and the ECM destabilization during joint degradatio

Interleukin-15 and interferon-γ participate in the cross-talk between natural killer and monocytic cells required for tumour necrosis factor production

We have characterized the lymphocyte subset and the receptor molecules involved in inducing the secretion of TNF by monocytic cells in vitro. The TNF secreted by monocytic cells was measured when they were co-cultured with either resting or IL-15-stimulated lymphocytes, T cells, B cells or natural killer (NK) cells isolated from the peripheral blood of healthy subjects and from the synovial fluid from patients with inflammatory arthropathies. Co-culture with IL-15-activated peripheral blood or synovial fluid lymphocytes induced TNF production by monocytic cells within 24 hours, an effect that was mainly mediated by NK cells. In turn, monocytic cells induced CD69 expression and IFN-γ production in NK cells, an effect that was mediated mainly by β(2 )integrins and membrane-bound IL-15. Furthermore, IFN-γ increased the production of membrane-bound IL-15 in monocytic cells. Blockade of β(2 )integrins and membrane-bound IL-15 inhibited TNF production, whereas TNF synthesis increased in the presence of anti-CD48 and anti-CD244 (2B4) monoclonal antibodies. All these findings suggest that the cross-talk between NK cells and monocytes results in the sustained stimulation of TNF production. This phenomenon might be important in the pathogenesis of conditions such as rheumatoid arthritis in which the synthesis of TNF is enhanced

Effects of disease activity on lipoprotein levels in patients with early arthritis: can oxidized LDL cholesterol explain the lipid paradox theory?

Background An increased risk of cardiovascular (CV) complications has been described in patients with rheumatoid arthritis (RA). It is the result of the combined effect of classic CV risk factors and others that are specific to the disease. Methods We assessed data from 448 early arthritis (EA) patients: 79% women, age (median [p25-p75]) at onset: 55 [44?67] years and disease duration at study entry 5 [3?8] months; and 72% fulfilled the 1987 RA criteria at 2?years of follow-up. Rheumatoid factor was positive in 54% of patients and anti-citrullinated peptide antibodies in 50%. The follow-up of patients ranged from 2 to 5?years with more than 1400 visits with lipoprotein measurements available (mean 2.5 visits/patient). Demographic- and disease-related variables were systematically recorded. Total cholesterol (TC), high-density lipoprotein (HDL-C), and low-density lipoprotein (LDL-C) levels were obtained from routine laboratory tests. Oxidized-LDL (oxLDL-C) levels were assessed using a commercial ELISA kit. We fitted population-averaged models nested by patient and visit to determine the effect of independent variables on serum levels of TC, its fractions, and oxLDL-C. Results After adjustment for several confounders, high-disease activity was significantly associated with decreased TC, HDL-C, and LDL-C levels and increased oxLDL-C levels. Standardized coefficients showed that the effect of disease activity was greater on oxLDL-C and HDL-C. Interestingly, we observed that those patients with lower levels of LDL-C showed higher oxLDL-C/LDL-C ratios. Conclusions High-disease activity in EA patients results in changes in the HDL-C and oxLDL-C levels, which in turn may contribute to the increased risk of CV disease observed in these patients.Our manuscript was supported by grants RD16/0011/0012, RD16/0011/0009, RD16/0011/0004, PI05/2044, and PI18/0371 from the Ministerio de Economía y Competitividad (Instituto de Salud Carlos III) and co-funded by Fondo Europeo de Desarrollo Regional (FEDER)

Experience with the use of Rituximab for the treatment of rheumatoid arthritis in a tertiary Hospital in Spain: RITAR study

There is evidence supporting that there are no relevant clinical differences between dosing rituximab 1000 mg or 2000 mg per cycle in rheumatoid arthritis (RA) patients in clinical trials, and low-dose cycles seem to have a better safety profile. Our objective was to describe the pattern of use of rituximab in real-life practice conditions. Methods: Rituximab for RA in clinical practice (RITAR) study is a retrospective cohort study from 2005 to 2015. Eligibility criteria were RA adults treated with rituximab for active articular disease. Response duration was the main outcome defined as months elapsed from the date of rituximab first infusion to the date of flare. A multivariable analysis was performed to determine the variables associated with response duration. Results: A total of 114 patients and 409 cycles were described, 93.0% seropositive and 80.7% women. Rituximab was mainly used as second-line biological therapy. On demand retreatment was used in 94.6% of cases versus fixed 6 months retreatment in 5.4%. Median response duration to on demand rituximab cycles was 10 months (interquartile range, 7–13). Multivariable analysis showed that age older than 65 years, number of rituximab cycles, seropositivity, and first- or second-line therapy were associated with longer response duration. The dose administered at each cycle was not significantly associated with response duration. Conclusions: Our experience suggests that 1000 mg rituximab single infusion on demand is a reasonable schedule for long-term treatment of those patients with good response after the first cycles, especially in seropositive patients and when it is applied as a first- or second-line biological therap

Assessing the influence of isotopic composition of water on that of clay minerals during chemical treatments.

The isotopic composition of hydrogen in authigenic minerals is a useful tool to reconstruct past paleo-environments. Clay minerals are an important component of authigenic minerals in soils and sediments but they usually occur with other compounds that must be eliminated before the analysis, such as organic matter and carbonates. Thus, various “pre-treatments” are used, generally involving dilute HCl and H2O2 solutions in water. In this work, the influence of the isotopic composition of the water used in these pre-treatment solutions is assessed, using ten different samples of clay minerals. The isotopic composition of hydrogen was measured in each sample after HCl pre-treatment alone, H2O2 pre-treatment alone and both HCl and H2O2 pre-treatments in sequence, using two types of water in the pre-treatment solutions: one 2H-enriched and one 2H-depleted. The results indicate some influence of the isotopic composition of the water on the clay minerals after pre-treatment. In general, the samples showed significant alteration by HCl pre-treatment and negligible alteration by H2O2 pre-treatment. A pure kaolinite reference material did not show any change by chemical pre-treatment while a smectite reference material did show significant effects. Other samples (Ethiopian lacustrine sediment samples and Spanish cave sediments) showed important differences, which also depend on clay mineralogy. Thus, mineralogy seems to be the main cause of the variability in the alteration, especially the quantity of smectite. In addition, this result challenges the utility of clay minerals for isotope studies in acid conditions, such as in acidic soils, to reconstruct past environments and, therefore, climate changes.This study was developed during a stay supported by predoctoral mobility grant from the Spanish MINECO. Financial support for this work was obtained from MINECO Grant CGL2015-65387-C3-3-P. I. Campaña was the beneficiary of a predoctoral FPI Grant from the Spanish MINECO. We thank Jessica Wilson at the University of South Florida School of Geosciences who supported the isotopic analysis. J.G. Wynn was supported by an NSF IR/D program. Special thanks to the local resident of Burgos that help us remove the car from a mud puddle. // Funding for open access charge: Universidad de Málaga / CBU

Improved classification of rheumatoid arthritis with a score including anti‑acetylated ornithine antibodies

The presence of rheumatoid factor (RF) or anti-cyclic citrullinated peptide (anti-CCP) autoantibodies contributes to the current rheumatoid arthritis (RA) classification criteria. These criteria involve stratification on antibody levels, which limits reproducibility, and underperform in the RA patients without RF and anti-CCP. Here, we have explored if two anti-acetylated peptide antibodies (AAPA), anti-acetylated lysine (AcLys) and anti-acetylated ornithine (AcOrn), could improve the performance of the current criteria. The analysis was done in 1062 prospectively-followed early arthritis (EA) patients. The anti-AcOrn were more informative than the anti-AcLys, the conventional RA antibodies and the anti-carbamylated protein antibodies. The anti-AcOrn produced a classification that did not require antibody levels and showed improved specificity (77.6% vs. 72.6%, p = 0.003) and accuracy (79.0% vs. 75.8%, p = 0.002) over the current criteria. These improvements were obtained with a scoring system that values concordance between anti-AcOrn, RF and anti-CCP. No significant gain was obtained in sensitivity (80.2% vs. 78.8%, p = 0.25) or in improving the classification of the RA patients lacking RF and anti-CCP, although the anti-AcOrn ranked first among the analysed new antibodies. Therefore, the anti-AcOrn antibodies could contribute to the improvement of RA classification criteria by exploiting antibody concordance.This work was supported by the Instituto de Salud Carlos III (Spain) through grants [RD16/0012/0014 and PI17/01606 to AG; RD16/0012/0012 to AB; PI14/00442 and RD16/0012/0011 to IG-A]. These grants are partially financed by the European Regional Development Fund of the EU (FEDER). LRM was supported by Xunta de Galicia (Spain) through a Gain pre-doctoral fellowship. CR was supported by Ministerio de Educacion Cultura y Deporte (Spain) through a FPU pre-doctoral fellowship [FPU15/03434]

Revision of TD1 and TD2 stratigraphic sequence of Gran Dolina cave (Sierra de Atapuerca, Spain)

Gran Dolina es una cueva rellena por al menos 25 m de sedimentos pleistocenos dividido en 12 unidades litoestratigráficas y 19 facies sedimentarias. Estas facies sedimentarias se han dividido entre facies alóctonas, definidas como entradas de sedimentos desde el exterior, y facies autóctonas, definidas como sedimentos generadas dentro del karst; sin embargo, esta clasificación de facies ha sido cuestionada en trabajos recientes. En este estudio se ha descrito en detalle las unidades TD1 y TD2 de Gran Dolina y se ha evaluado la idoneidad del uso de facies autóctonas. Para ello, se ha estudiado la sección estratigráfica de la excavación arqueológica, combinando observaciones de campo con análisis de laboratorio (tamizado, difracción láser y DRX) para caracterizar la textura y estructura de los sedimentos. A partir de estos estudios, se han identificado un total de 8 facies sedimentarias, y se han separado la unidad TD1 en dos sub-unidades y 13 niveles, y la unidad TD2 en tres sub-unidades. La asociación de facies indica una sucesión de fases freáticas y vadosas que definiría conjuntamente condiciones epifreáticas en el interior de la cueva, relacionadas con la transición entre la terraza T3 y la terraza T4 del valle del río Arlanzón. Por tanto, se propone el término facies de interior (y facies de entrada en vez de facies alóctonas) para definir los sedimentos de las unidades de TD1 y TD2 de Gran Dolina, ya que el análisis de facies indica transporte de los sedimentos por corrientes subterráneas.This study was supported by the project PGC2018-093925-B-C31 of the Spanish Ministry of Science and Innovation, Spanish State Research Agency and FEDER funds from the UE. Project PID2021-122355NB-C33 financed by MCIN/AEI/10.13039/501100011033/FEDER, UE. Funding for open access charge: Universidad de Málaga/CBUA. I. Campaña is the beneficiary of a postdoctoral grant from Junta de Andalucía. This work has benefited from discussion with Mathieu Duval, Lucía Bermejo, Lucía Ojeda and Sergio Durán. The constructive comments made by one anonymous reviewer and Dr. Ivan Martini contributed to improve the manuscript

An Overview of VPAC Receptors in Rheumatoid Arthritis: Biological Role and Clinical Significance

The axis comprised by the Vasoactive Intestinal Peptide (VIP) and its G protein-coupled receptors (GPCRs), VPAC1, and VPAC2, belong to the B1 family and signal through Gs or Gq proteins. VPAC receptors seem to preferentially interact with Gs in inflammatory cells, rather than Gq, thereby stimulating adenylate cyclase activity. cAMP is able to trigger various downstream pathways, mainly the canonical PKA pathway and the non-canonical cAMP-activated guanine nucleotide exchange factor (EPAC) pathway. Classically, the presence of VPACs has been confined to the plasma membrane; however, VPAC1 location has been described in the nuclear membrane in several cell types such as activated Th cells, where they are also functional. VPAC receptor signaling modulates a number of biological processes by tipping the balance of inflammatory mediators in macrophages and other innate immune cells, modifying the expression of TLRs, and inhibiting MMPs and the expression of adhesion molecules. Receptor signaling also downregulates coagulation factors and acute-phase proteins, promotes Th2 over Th1, stimulates Treg abundance, and finally inhibits a pathogenic Th17 profile. Thus, the VIP axis signaling regulates both the innate and adaptive immune responses in several inflammatory/autoimmune diseases. Rheumatoid arthritis (RA) is a complex autoimmune disease that develops on a substrate of genetically susceptible individuals and under the influence of environmental factors, as well as epigenetic mechanisms. It is a heterogeneous disease with different pathogenic mechanisms and variable clinical forms between patients with the same diagnosis. The knowledge of VIP signaling generated in both animal models and human ex vivo studies can potentially be translated to clinical reality. Most recently, the beneficial effects of nanoparticles of VIP self-associated with sterically stabilized micelles have been reported in a murine model of RA. Another novel research area is beginning to define the receptors as biomarkers in RA, with their expression levels shown to be associated with the activity of the disease and patients-reported impairment. Therefore, VPAC expression together VIP genetic variants could allow patients to be stratified at the beginning of the disease with the purpose of guiding personalized treatment decisions

Lack of association between TLR4 rs4986790 polymorphism and risk of cardiovascular disease in patients with rheumatoid arthritis

This is copy of an article published in the DNA and cell biology 2012 © Mary Ann Liebert, Inc.; DNA and cell bilogy is available online at: http://online.liebertpub.comRheumatoid arthritis (RA) is a chronic inflammatory disease associated with increased cardiovascular (CV) mortality. Toll-like receptor-4 (TLR4) activates the innate immune response via NF-kB pathway and mitogenactivated protein kinase signaling, leading to expression of proinflammatory cytokines and chemokines. The G allele of TLR4 rs4986790 (+ 896A > G, Asp299Gly) gene polymorphism has been implicated in reduction of risk of atherosclerosis. In this study, 1481 RA patients fulfilling the 1987 American College of Rheumatology (ACR) criteria were genotyped for the rs4986790 TLR4 variant to determine the influence of this variant in the risk of CV events in these patients. Also, HLA-DRB1 status was determined using molecular based methods. Moreover, potential influence of rs4986790 variant in the development of subclinical atherosclerosis was assessed in a subgroup of RA patients with no history of CV events by the measurement of surrogate markers of subclinical atherosclerosis. No statistically significant differences in allele or genotype frequencies for the rs4986790 variant between RA patients who experienced CV events or not were found. Likewise, no significant association between this gene variant and any of the surrogate markers of subclinical atherosclerosis was found. In summary, results in our study do not support the hypothesis that the rs4986790 (+ 896A > G, Asp299Gly) TLR4 variant may influence predisposition for subclinical atherosclerosis and clinically evident CV disease in RA patientsThis study was supported by two grants from Fondo de Investigaciones Sanitarias PI06-0024 and PS09/00748 (Spain). This work was partially supported by RETICS Program, RD08/0075 (RIER) from Instituto de Salud Carlos III (ISCIII), within the VI PN de I +D+ i 2008–2011 (FEDER). M.G.B. is supported by a grant from Fundación Española de Reumatología (FER). R.L.M. is supported by a grant by IFIMAV, Santander (Spain)