Relapse rates in patients with rheumatoid arthritis in stable remission tapering or stopping antirheumatic therapy: interim results from the prospective randomised controlled RETRO study

Objective: To prospectively analyse the risk for disease relapses in patients with rheumatoid arthritis (RA) in sustained remission, either continuing, tapering or stopping disease-modifying antirheumatic drugs (DMARDs) in a prospective randomised controlled trial. Methods: Reduction of Therapy in patients with Rheumatoid arthritis in Ongoing remission is a multicentre, randomised controlled, parallel-group phase 3 trial evaluating the effects of tapering and stopping all conventional and/or biological DMARDs in patients with RA in stable remission. Patients (disease activity score 28 (DAS28)<2.6 for least 6 months) were randomised into three arms, either continuing DMARDs (arm 1), tapering DMARDs by 50% (arm 2) or stopping DMARDs after 6 months tapering (arm 3). The primary endpoint was sustained remission during 12 months. Results: In this interim analysis, the first 101 patients who completed the study were analysed. At baseline, all patients fulfilled DAS28 remission and 70% also American College of Rheumatology- European League Against Rheumatism Boolean remission. 82.2% of the patients received methotrexate, 40.6% biological DMARDs and 9.9% other DMARDs. Overall, 67 patients (66.3%) remained in remission for 12 months, whereas 34 patients (33.7%) relapsed. The incidence of relapses was related to study arms (p=0.007; arm 1: 15.8%; arm 2: 38.9%; arm 3: 51.9%). Multivariate logistic regression identified anticitrullinated protein antibodies (ACPA) positivity (p=0.038) and treatment reduction (in comparison to continuation) as predictors for relapse (arm 2: p=0.012; arm 3: p=0.003). Conclusions: This randomised controlled study testing three different treatment strategies in patients with RA in sustained remission demonstrated that more than half of the patients maintain in remission after tapering or stopping conventional and biological DMARD treatment. Relapses occurred particularly in the first 6 months after treatment reduction and were associated with the presence of ACPA. Trial registration number: 2009-015740-42

Prediction of disease relapses by multibiomarker disease activity and autoantibody status in patients with rheumatoid arthritis on tapering DMARD treatment

Objective: To analyse the role of multibiomarker disease activity (MBDA) score in predicting disease relapses in patients with rheumatoid arthritis (RA) in sustained remission who tapered disease modifying antirheumatic drug (DMARD) therapy in RETRO, a prospective randomised controlled trial. Methods: MBDA scores (scale 1–100) were determined based on 12 inflammation markers in baseline serum samples from 94 patients of the RETRO study. MBDA scores were compared between patients relapsing or remaining in remission when tapering DMARDs. Demographic and disease-specific parameters were included in multivariate logistic regression analysis for defining predictors of relapse. Results: Moderate-to-high MBDA scores were found in 33% of patients with RA overall. Twice as many patients who relapsed (58%) had moderate/high MBDA compared with patients who remained in remission (21%). Baseline MBDA scores were significantly higher in patients with RA who were relapsing than those remaining in stable remission (N=94; p=0.0001) and those tapering/stopping (N=59; p=0.0001). Multivariate regression analysis identified MBDA scores as independent predictor for relapses in addition to anticitrullinated protein antibody (ACPA) status. Relapse rates were low (13%) in patients who were MBDA−/ACPA−, moderate in patients who were MBDA+/ACPA− (33.3%) and MBDA−ACPA+ (31.8%) and high in patients who were MBDA+/ACPA+ (76.4%). Conclusions: MBDA improved the prediction of relapses in patients with RA in stable remission undergoing DMARD tapering. If combined with ACPA testing, MBDA allowed prediction of relapse in more than 80% of the patients. Trial registration number: EudraCT 2009-015740-42

Putting cells under pressure ::a simple and efficient way to enhance the productivity of medium-chain-length polyhydroxyalkanoate in processes with Pseudomonas putida KT2440

The success of bioprocess implementation relies on the ability to achieve high volumetric productivities and requires working with high-cell-density cultivations. Elevated atmospheric pressure might constitute a promising tool for enhancing the oxygen transfer rate (OTR), the major growth-limiting factor for such cultivations. However, elevated pressure and its effects on the cellular environment also represent a potential source of stress for bacteria and may have negative effects on product formation. In order to determine whether elevated pressure can be applied for enhancing productivity in the case of medium-chain-length polyhydroxyalkanoate (mcl-PHA) production by Pseudomonas putida KT2440, the impact of a pressure of 7 bar on the cell physiology was assessed. It was established that cell growth was not inhibited by this pressure if dissolved oxygen tension (DOT) and dissolved carbon dioxide tension (DCT) were kept below ∼30 and ∼90 mg L−1, respectively. Remarkably, a little increase of mcl-PHA volumetric productivity was observed under elevated pressure. Furthermore, the effect of DCT, which can reach substantial levels during high-cell-density processes run under elevated pressure, was investigated on cell physiology. A negative effect on product formation could be dismissed since no significant reduction of mcl-PHA content occurred up to a DCT of ∼540 mg L−1. However, specific growth rate exhibited a significant decrease, indicating that successful high-cell-density processes under elevated pressure would be restricted to chemostats with low dilution rates and fed-batches with a small growth rate imposed during the final part. This study revealed that elevated pressure is an adequate and efficient way to enhance OTR and mcl-PHA productivity. We estimate that the oxygen provided to the culture broth under elevated pressure would be sufficient to triple mcl-PHA productivity in our chemostat system from 3.4 (at 1 bar) to 11 g L−1 h−1 (at 3.2 bar)

New insights on the reorganization of gene transcription in Pseudomonas putida KT2440 at elevated pressure

Abstract Background Elevated pressure, elevated oxygen tension (DOT) and elevated carbon dioxide tension (DCT) are readily encountered at the bottom of large industrial bioreactors and during bioprocesses where pressure is applied for enhancing the oxygen transfer. Yet information about their effect on bacteria and on the gene expression thereof is scarce. To shed light on the cellular functions affected by these specific environmental conditions, the transcriptome of Pseudomonas putida KT2440, a bacterium of great relevance for the production of medium-chain-length polyhydroxyalkanoates, was thoroughly investigated using DNA microarrays. Results Very well defined chemostat cultivations were carried out with P. putida to produce high quality RNA samples and ensure that differential gene expression was caused exclusively by changes of pressure, DOT and/or DCT. Cellular stress was detected at 7 bar and elevated DCT in the form of heat shock and oxidative stress-like responses, and indicators of cell envelope perturbations were identified as well.Globally, gene transcription was not considerably altered when DOT was increased from 40 ± 5 to 235 ± 20% at 7 bar and elevated DCT. Nevertheless, differential transcription was observed for a few genes linked to iron-sulfur cluster assembly, terminal oxidases, glutamate metabolism and arginine deiminase pathway, which shows their particular sensitivity to variations of DOT. Conclusions This study provides a comprehensive overview on the changes occurring in the transcriptome of P. putida upon mild variations of pressure, DOT and DCT. Interestingly, whereas the changes of gene transcription were widespread, the cell physiology was hardly affected, which illustrates how efficient reorganization of the gene transcription is for dealing with environmental changes that may otherwise be harmful. Several particularly sensitive cellular functions were identified, which will certainly contribute to the understanding of the mechanisms involved in stress sensing/response and to finding ways of enhancing the stress tolerance of microorganisms.This work was partially supported by the Swiss National Science Foundation grant 315200-116812/1 and by the Spanish grant BIO2010-21049 from the Comisión Interministerial de Ciencia y Tecnología.Peer Reviewe

Deciphering the Phytochemical Profile of an Alpine Rose (<i>Rhododendron ferrugineum</i> L.) Leaf Extract for a Better Understanding of Its Senolytic and Skin-Rejuvenation Effects

Rhododendron ferrugineum, commonly named Alpine rose, is an emblematic medicinal plant of European mountains. In this study, the chemical profile of a glycerol/water extract developed from this plant as a cosmetic ingredient is investigated to understand the extract constituent(s) that could mostly contribute to its senolytic activity and skin-rejuvenation effects. For this purpose, the dereplication method “CARAMEL”, which combines Centrifugal Partition Chromatography to Nuclear Magnetic Resonance data interpretation, was directly applied to the hydro-glycerinated extract, leading to the unambiguous identification of fourteen Alpine rose metabolites, despite the strong presence of the heavy solvent glycerol. Flavonoids derived from taxifolin, quercetin, and (+)-catechin were identified as significant constituents of the extract, followed by flavanones, orcinol derivatives, phloroacetophenone, and phenolic acids, as well as the pentacyclic triterpene lupeol. Given that senolytic molecules are known to selectively induce the death of senescent cells without affecting healthy proliferating cells, which can be achieved by the selective inhibition or downregulation of the anti-apoptotic Bcl-2 protein, and considering the well-recognized pro-apoptotic activity of hyperoside, taxifolin, naringenin and farrerol, the senolytic activity of the glycerol/water Alpine rose extract can be explained by the abundance of flavonoids present in the extract

Association of the ANCA-associated vasculitis (AAV) patient-reported outcome (AAV-PRO) questionnaire with established outcome measures in AAV

Objectives The ANCA-associated vasculitis (AAV) patient-reported outcome (AAV-PRO) questionnaire was developed to capture the impact of AAV and its treatment. We investigated the association of specific AAV-PRO domains with disease activity and extent, damage, depression, health-related quality of life and treatment. Methods In a prospective longitudinal study AAV-PRO, Beck’s depression inventory (BDI), Short Form 36 (SF36), BVAS and Vasculitis Damage Index (VDI) were completed at baseline (t1) and after 3–6 months (t2). In addition, patient data including diagnosis, therapies, relapses, and organ manifestations were recorded. Data were analyzed by t-tests and correlation-based regression analyses. Results 156 patients with AAV participated. The mean BVAS at the time of enrolment was 1.4 ± 3.74. Median AAV-PRO domain scores were higher in patients reporting “active disease” compared with patients reporting “in remission” (p&amp;lt; 0.001). In the correlation analyses all AAV-PRO domain scores correlated strongly with the BDI (all r ≥ 0.319, all p≤ 0.001) as well as all eight SF36 subdomains (all |r|≥0.267, all p≤ 0.001). The regression analyses showed that AAV-PRO domains were strongly predicted by BDI and SF36 domains (|β|≥0.240 for the strongest predictor of each domain). In the longitudinal comparison (t1/t2) there were no significant changes of the overall results. Conclusion Our data show convergent validity of all AAV-PRO subdomains with the established questionnaires BDI and SF-36. The AAV-PRO domains scores were not correlated with clinician derived instruments including the BVAS and VDI. Thus, we regard the AAV-PRO as a valuable addition that might complement traditional endpoints in clinical trials