Search

Central nervous system (CNS) barrier impairment has been reported in amyotrophic lateral sclerosis (ALS), highlighting its potential significance in the disease. In this context, we aim to shed light on its involvement in the disease, by determining albumin quotient (QAlb) at the time of diagnosis of ALS in a large cohort of patients. Patients from the university hospital of Tours (n = 307) were included in this monocentric, retrospective study. In total, 92 patients (30%) had elevated QAlb levels. This percentage was higher in males (43%) than in females (15%). Interestingly, QAlb was not associated with age of onset, age at sampling or diagnostic delay. However, we found an association with ALS functional rating scale-revised (ALSFRS-r) at diagnosis but this was significant only in males. The QAlb levels were not linked to the presence of a pathogenic mutation. Finally, we performed a multivariate survival analysis and found that QAlb was significantly associated with survival in male patients (HR = 2.3, 95% CI = 1.2–4.3, p = 0.009). A longitudinal evaluation of markers of barrier impairment, in combination with inflammatory biomarkers, could give insight into the involvement of CNS barrier impairment in the pathogenesis of the disease. The gender difference might guide the development of new drugs and help personalise the treatment of ALS.

Iron metabolism is tightly linked to infectious and inflammatory signals through hepcidin synthesis. To date, iron homeostasis during SARS-CoV-2 infection has not yet been described. The aim of this study is to characterize the hepcidin and erythroid regulators (growth differentiation factor 15 (GDF-15) and erythroferrone (ERFE)) by measuring concentrations in plasma in context of COVID-19 disease. We performed a single-center observational study of patients with COVID-19 to evaluate concentrations of main regulatory proteins involved in iron homeostasis, namely: hepcidin, ERFE and GDF-15. SARS-CoV-2 infection (COVID-19+) was defined by a positive RT-PCR. Sixteen patients with COVID-19+ were gender-matched and age-matched to 16 patients with a sepsis unrelated to SARS-CoV-2 (COVID-19−) and were compared with non-parametric statistic test. Clinical and hematological parameters, plasma iron, transferrin, transferrin saturation, ferritin, soluble transferrin receptor and C reactive protein were not statistically different between both groups. Median plasma hepcidin concentrations were higher in the COVID-19+ group (44.1 (IQR 16.55–70.48) vs 14.2 (IQR 5.95–18.98) nmol/L, p=0.003), while median ERFE and GDF-15 concentrations were lower in the COVID-19+ group (0.16 (IQR 0.01–0.73) vs 0.89 (IQR 0.19–3.82) ng/mL, p=0.035; 2003 (IQR 1355–2447) vs 4713 (IQR 2082–7774) pg/mL, p=0015), respectively) compared with the COVID-19− group. This is the first study reporting lower ERFE and GDF-15 median concentrations in patients with COVID-19+ compared with patients with COVID-19−, associated with an increased median concentration of hepcidin in the COVID-19+ group compared with COVID19− group.

A 16-year-old child with no medical history was admitted to the hospital emergency for abdominal pain associated with polyuria-polydipsia and weight loss (baseline BMI: 25,4 kg/m2). Diagnosis of severe ketoacidosis was quickly raised regarding major metabolic acidosis, high ketonemia and glycemia. Acute pancreatitis was then diagnosed according to a plasmatic lipase more than tenfold normal values associated with a severe hypertriglyceridemia superior to 100 mmol/L. The triad composed of diabetic ketoacidosis-acute pancreatitis-hypertriglyceridemia is rarely found in childhood and can have deleterious consequences. The etiology of this disease is still enigmatic, as one can be both, cause and consequence of the other. Genetic investigation of familial chylomicronemia legitimated to invalidate the dyslipidemia etiology of this event. On the other hand, the association of a genetic variant of lipoprotein lipase leading to a decrease in its activity, with the insulinopenia of type 1 diabetes most certainly triggered this episode of hypertriglyceridemia.Une jeune adolescente de 16 ans, sans antécédent médical, s'est présentée aux urgences pour douleurs abdominales dans un contexte de polyuro-polydipsie avec amaigrissement (IMC initial : 25,4 kg/m2). Une acidocétose sévère a rapidement été évoquée devant une acidose métabolique majeure, ainsi qu'une cétonémie et glycémie élevées. Une pancréatite aiguë a ensuite été diagnostiquée devant une lipase plasmatique supérieure à 10 fois les valeurs normales associée à une hypertriglycéridémie majeure de plus de 100 mmol/L. La triade acidocétose-pancréatite aiguë-hypertriglycéridémie est un phénomène très rarement retrouvé dans l'enfance et qui peut avoir des conséquences dramatiques. Il s'agit d'une pathologie à l'étiologie encore énigmatique, l'une pouvant être la cause et la conséquence de l'autre. L'exploration génétique d'une hyperchylomicronémie a pu permettre d'infirmer l'étiologie dyslipidémique de cet épisode. En revanche, l'association d'un variant génétique de la lipoprotéine lipase conduisant à une diminution de son activité, à l'insulinopénie du diabète de type 1 a très certainement déclenché cet épisode d'hypertriglycéridémie.

International audience; The pathophysiology of primary burning mouth syndrome (BMS) remains controversial. Targeted analyses or "omics" approach of saliva provide diagnostic or pathophysiological biomarkers. This pilot study's primary objective was to explore the pathophysiology of BMS through a comparative analysis of the salivary metabolome among 26 BMS female cases and 25 age- and sex-matched control subjects. Secondary objectives included comparative analyses of inflammatory cytokines, neuroinflammatory markers, and steroid hormones among cases and control subjects, and among BMS patients according to their clinical characteristics. Salivary metabolome, neuroinflammatory markers, cytokines, and steroids were, respectively, analysed by liquid chromatography coupled with mass spectrometry, ELISA and protease activity assay, and multiparametric Luminex method. Among the 166 detected metabolites, univariate analysis did not find any discriminant metabolite between groups. Supervised multivariate analysis divided patients into 2 groups with an accuracy of 60% but did not allow significant discrimination (permutation test, P = 0.35). Among the metabolites contributing to the model, 3 belonging to the tyrosine pathway ( l -dopa, l -tyrosine, and tyramine) were involved in the discrimination between cases and control subjects, and among BMS patients according to their levels of pain. Among the detectable molecules, levels of cytokines, steroid hormones, and neuroinflammatory markers did not differ between cases and control subjects and were not associated with characteristics of BMS patients. These results do not support the involvement of steroid hormones, inflammatory cytokines, or inflammatory neurogenic mediators in the pathophysiology of pain in BMS, whereas the observed shift in tyrosine metabolism may indicate an adaptative response to chronic pain or an impaired dopaminergic transmission.

International audience; Chronic stimulation by infectious pathogens or self-antigen glucosylsphingosine (GlcSph) can lead to monoclonal gammopathy of undetermined significance (MGUS) and multiple myeloma (MM). Novel assays such as the multiplex infectious antigen microarray (MIAA) and GlcSph assays, permit identification of targets for >60% purified monoclonal immunoglobulins (Igs). Searching for additional targets, we selected 28 purified monoclonal Igs whose antigen was not represented on the MIAA and GlcSph assays; their specificity of recognition was then analyzed using microarrays consisting of 3760 B-cell epitopes from 196 pathogens. The peptide sequences PALTAVETG and PALTAAETG of the VP1 coat proteins of human poliovirus 1/3 and coxsackievirus B1/B3, respectively, were specifically recognized by 6/28 monoclonal Igs. Re-analysis of patient cohorts showed that purified monoclonal Igs from 10/155 MGUS/SM (6.5%) and 3/147 MM (2.0%) bound to the PALTAVETG or PALTAAETG epitopes. Altogether, PALTAV/AETG-initiated MGUS are not rare and few seem to evolve toward myeloma.

HIV-1 assembly occurs principally at the plasma membrane (PM) of infected cells. Gag polyprotein precursors (Pr55(Gag)) are targeted to the PM, and their binding is mediated by the interaction of myristoylated matrix domain and a PM-specific phosphoinositide, the phosphatidylinositol-(4,5)-bisphosphate [PI(4,5)P(2)]. The major synthesis pathway of PI(4,5)P(2) involves the activity of phosphatidylinositol-4-phosphate 5-kinase family type 1 composed of three isoforms (PIP5K1α, PIP5K1β, and PIP5K1γ). To examine whether the activity of a specific PIP5K1 isoform determines proper Pr55(Gag) localization at the PM, we compared the cellular behavior of Pr55(Gag) in the context of PIP5K1 inhibition using siRNAs that individually targeted each of the three isoforms in TZM-bl HeLa cells. We found that downregulation of PIP5K1α and PIP5K1γ strongly impaired the targeting of Pr55(Gag) to the PM with a rerouting of the polyprotein within intracellular compartments. The efficiency of Pr55(Gag) release was thus impaired through the silencing of these two isoforms, while PIP5K1β is dispensable for Pr55(Gag) targeting to the PM. The PM mistargeting due to the silencing of PIP5K1α leads to Pr55(Gag) hydrolysis through lysosome and proteasome pathways, while the silencing of PIP5K1γ leads to Pr55(Gag) accumulation in late endosomes. Our findings demonstrated that, within the PIP5K1 family, only the PI(4,5)P(2) pools produced by PIP5K1α and PIP5K1γ are involved in the Pr55(Gag) PM targeting process. IMPORTANCE PM specificity of Pr55(Gag) membrane binding is mediated through the interaction of PI(4,5)P(2) with the matrix (MA) basic residues. It was shown that overexpression of a PI(4,5)P(2)-depleting enzyme strongly impaired PM localization of Pr55(Gag). However, cellular factors that control PI(4,5)P(2) production required for Pr55(Gag)-PM targeting have not yet been characterized. In this study, by individually inhibiting PIP5K1 isoforms, we elucidated a correlation between PI(4,5)P(2) metabolism pathways mediated by PIP5K1 isoforms and the targeting of Pr55(Gag) to the PM of TZM-bl HeLa cells. Confocal microscopy analyses of cells depleted from PIP5K1α and PIP5K1γ show a rerouting of Pr55(Gag) to various intracellular compartments. Notably, Pr55(Gag) is degraded by the proteasome and/or by the lysosomes in PIP5K1α-depleted cells, while Pr55(Gag) is targeted to endosomal vesicles in PIP5K1γ-depleted cells. Thus, our results highlight, for the first time, the roles of PIP5K1α and PIP5K1γ as determinants of Pr55(Gag) targeting to the PM.

ObjectivesAnti-drug antibodies (ADA) are responsible for decreased adalimumab efficacy in axial spondyloarthritis (SpA). We aimed to evaluate the ability of methotrexate (MTX) to decrease adalimumab immunisation.MethodsA total of 110 patients eligible to receive adalimumab 40 mg subcutaneously (s.c.) every other week were randomised (1:1 ratio) to receive, 2 weeks before adalimumab (W-2) and weekly, MTX 10 mg s.c. (MTX+) or not (MTX−). ADA detection and adalimumab serum concentration were assessed at weeks 4 (W4), 8 (W8), 12 (W12) and 26 (W26) after starting adalimumab (W0). The primary outcome was the proportion of patients with ADA at W26. Four years after the study completion, we retrospectively analysed adalimumab maintenance in relation with MTX co-treatment duration.ResultsWe analysed data for 107 patients (MTX+; n=52; MTX-; n=55). ADA were detected at W26 in 39/107 (36.4%) patients: 13/52 (25%) in the MTX+ group and 26/55 (47.3%) in the MTX− group (p=0.03). Adalimumab concentration was significantly higher in the MTX+ than MTX− group at W4, W8, W12 and W26. The two groups did not differ in adverse events or efficacy. In the follow-up study, MTX co-treatment >W26 versus no MTX or ≤W26 was significantly associated with adalimumab long-term maintenance (p=0.04).ConclusionMTX reduces the immunogenicity and ameliorate the pharmacokinetics of adalimumab in axial SpA. A prolonged co-treatment of MTX>W26 seems to increase adalimumab long-term maintenance.

HIV-1 assembly occurs principally at the plasma membrane (PM) of infected cells. Gag polyprotein precursors (Pr55Gag) are targeted to the PM, and their binding is mediated by the interaction of myristoylated matrix domain and a PM-specific phosphoinositide, the phosphatidylinositol-(4,5)-bisphosphate [PI(4,5)P2]. The major synthesis pathway of PI(4,5)P2 involves the activity of phosphatidylinositol-4-phosphate 5-kinase family type 1 composed of three isoforms (PIP5K1α, PIP5K1β, and PIP5K1γ). To examine whether the activity of a specific PIP5K1 isoform determines proper Pr55Gag localization at the PM, we compared the cellular behavior of Pr55Gag in the context of PIP5K1 inhibition using siRNAs that individually targeted each of the three isoforms in TZM-bl HeLa cells. We found that downregulation of PIP5K1α and PIP5K1γ strongly impaired the targeting of Pr55Gag to the PM with a rerouting of the polyprotein within intracellular compartments. The efficiency of Pr55Gag release was thus impaired through the silencing of these two isoforms, while PIP5K1β is dispensable for Pr55Gag targeting to the PM. The PM mistargeting due to the silencing of PIP5K1α leads to Pr55Gag hydrolysis through lysosome and proteasome pathways, while the silencing of PIP5K1γ leads to Pr55Gag accumulation in late endosomes. Our findings demonstrated that, within the PIP5K1 family, only the PI(4,5)P2 pools produced by PIP5K1α and PIP5K1γ are involved in the Pr55Gag PM targeting process.IMPORTANCE PM specificity of Pr55Gag membrane binding is mediated through the interaction of PI(4,5)P2 with the matrix (MA) basic residues. It was shown that overexpression of a PI(4,5)P2-depleting enzyme strongly impaired PM localization of Pr55Gag However, cellular factors that control PI(4,5)P2 production required for Pr55Gag-PM targeting have not yet been characterized. In this study, by individually inhibiting PIP5K1 isoforms, we elucidated a correlation between PI(4,5)P2 metabolism pathways mediated by PIP5K1 isoforms and the targeting of Pr55Gag to the PM of TZM-bl HeLa cells. Confocal microscopy analyses of cells depleted from PIP5K1α and PIP5K1γ show a rerouting of Pr55Gag to various intracellular compartments. Notably, Pr55Gag is degraded by the proteasome and/or by the lysosomes in PIP5K1α-depleted cells, while Pr55Gag is targeted to endosomal vesicles in PIP5K1γ-depleted cells. Thus, our results highlight, for the first time, the roles of PIP5K1α and PIP5K1γ as determinants of Pr55Gag targeting to the PM.

International audience

Background Solitary plasmacytoma of the liver is a very rare and aggressive form of plasma cell dyscrasia. To the best of our knowledge, very few cases have been reported without systemic disease. We reported a rare case of hepatic solitary plasmacytoma that successfully responded to fractionated stereotactic radiotherapy. Case presentation A 64-year-old white French man had monoclonal gammopathy of the immune globulin G lambda type; he developed a cholestasis and cytolysis with the discovery of a subscapular nodule. A biopsy showed plasma cells and, for several reasons, the decision was made to use the fractionated stereotactic radiotherapy strategy. After 20 months, he is asymptomatic and the immune globulin G component has completely disappeared. Conclusion We suggest considering Cyberknife® radiosurgery as an option for the treatment of hepatic solitary plasmacytoma.

Aim Hepatocellular carcinoma (HCC) is a common outcome of chronic hepatitis C virus (HCV) infection and constitutes the main burden of this disease. The molecular mechanisms underlying the development of HCC are multiple and might involve certain microRNA (miR). Since discordant results have been reported concerning the detection of expression of miR-152 and miR-122 in HCC, our aim was to measure the levels of both miRs in serum and liver samples. Methods We analyzed miR-152 and miR-122 expression by RT-qPCR in a serum cohort from 14 HCV-infected patients who developed HCC, 20 HCV+ patients without HCC and 19 control patients. We also studied miR-152 and miR-122 in an independent tissue cohort from 11 normal livers, and from paired HCC and non-tumor adjacent livers of 11 HCV-infected patients and 12 non-infected patients. Results In serum samples, higher levels of miR-122 were found in non-HCC HCV+ compared to HCC HCV+ and control groups whereas miR-152 was detectable in a lower range in HCC HCV+ compared to non-HCC HCV+ and control groups. We found higher signals for miR-122 and miR-152 in non-tumor liver and HCC tissues compared to control tissues. HCC etiology had no detectable influence on miR-122 expression whereas miR-152 was increased in HCV+ tissue samples. Conclusions Detection of low values of circulating miR-152 is a potentially interesting marker of hepatocarcinogenesis in HCV+ patients, in contrast to miR-122, which varies according to hepatocyte damage.

International audience; Hepatitis C virus (HCV) virions contain a subset of host liver cells proteome often composed of interesting virus-interacting factors. A proteomic analysis performed on double gradient-purified clinical HCV highlighted the translation regulator LARP1 on these virions. This finding was validated using post-virion capture and immunoelectron microscopy, as well as immunoprecipitation applied to in vitro (Huh7.5 liver cells) grown (Gt2a, JFH1 strain) and patient-derived (Gt1a) HCV particles. Upon HCV infection of Huh7.5 cells, we observed a drastic transfer of LARP1 to lipid droplets, inducing colocalization with core proteins. RNAi-mediated depletion of LARP1 using the C911 control approach decreased extracellular infectivity of HCV Gt1a (H77), Gt2a (JFH1), and Gt3a (S52 chimeric strain), yet increased their intracellular infectivity. This latter effect was unrelated to changes in the hepatocyte secretory pathway, as evidenced using a functional RUSH assay. These results indicate that LARP1 binds to HCV, an event associated with retention of intracellular infectivity.

Background Adalimumab significantly reduces the activity of rheumatoid arthritis (RA), but reliable biomarkers of inflammation are still lacking to predict and evaluate the therapeutic response. Serum calprotectin is a mainstay of endogenous activation of the inflammatory response that can be useful as a marker of response to treatment in RA. Objectives To compare the evolution over time of serum calprotectin and C-Reactive Protein (CRP) after initiation of adalimumab. Methods Serum levels of calprotectin, CRP and adalimumab concentration were measured at visits V1 (week 0), V2 (week 4), V3 (week 8), V4 (week 12) and V5 (week 26). Changes in each variable were analyzed at each visit and each variable was compared to each other using a correlation test. Receiving operating characteristic curves were used to estimate the predictive value of response at V5 for calprotectin and CRP at each visit. Results Data from 66 patients were analyzed. Serum calprotectin level decreased from V1 to V5 (3.76 µg/mL [0 – 17.47] to 2.74 µg/mL [0 – 18.83]; p < 0.05). A positive correlation was observed between serum calprotectin and DAS28ESR (Spearman 0.244; p < 0.01), and between CRP and DAS28ESR (Spearman 0.512; p < 0.01) for all visits combined. In contrast to CRP, serum calprotectin and serum calprotectin variation from V1 at each visit, were not predictive of DAS28ESR at V5. Conclusion Serum calprotectin decreases after initiation of adalimumab in RA but was weakly correlated with disease activity at week 26. Serum calprotectin cannot be considered as superior to CRP as predictive marker of therapeutic response in RA after initiation of adalimumab. Disclosure of Interests Guillaume Servant: None declared, Christophe Passot: None declared, Eric Piver: None declared, Oscar Knight: None declared, Valerie Devauchelle-Pensec Grant/research support from: Roche-Chugai, Speakers bureau: MSD, BMS, UCB, Roche, Stephanie Rist: None declared, Aleth Perdriger: None declared, Elisabeth Gervais Speakers bureau: Abbvie, BMS, MSD, Pfzer, Roche, UCB, Novartis, Emmanuelle Dernis: None declared, Benoit Le Goff Speakers bureau: Abbvie, BMS, Janssen, MSD, Pfzer, Sanofi-Genzyme, UCB, Novartis, Laurence Picon: None declared, Philippe Goupille Grant/research support from: Financial compensation received from MSD on a pro-rota basis for participation in Scientific Committee meetings and functions for this study, Speakers bureau: Abbvie, Biogaran, BMS, Hospira, Janssen, MSD, Pfizer, Sanofi-Genzyme, UCB, Denis Mulleman Speakers bureau: Pfizer, Novartis, Grifols

Background: Immunogenicity of anti TNF monoclonal antibodies leads to poor or secondary loss of response. Methotrexate reduces anti-drug antibodies (ADA) to adalimumab at week 26 in spondyloarthritis (SpA).1 Objectives: Herein we sought to examine adalimumab long term persistence in aDA positive versus aDA negative SpA patients. Methods: The CoMARIS study (Combination of Methotrexate and adalimumab to Reduce Immunization in patients with axial SpA) is a 26-week prospective, randomised, open-labelled, multicentre study in which patients received adalimumab 40 mg subcutaneously (s.c.) every other week either in combination with MTX 10 mg s.c. For 26 weeks or without MTX. In a post hoc analysis, we reviewed the charts of patients to assess adalimumab persistence. A Cox model analysis was performed to test the following covariates: MTX combination or not, sex, presence of aDA at week 26. Results: Data from 104 patients (54 without MTX and 50 with MTX) were reviewed, and time of adalimumab discontinuation was collected. The median time of follow-up was 210.57 weeks. ADA positivity at week 26 was the only covariate associated with adalimumab persistence. The median retention rate of adalimumab in aDA positive patients was 56.9 weeks, as compared with 98.6 weeks in those without aDA (log rank: p=0.015). In the Cox model analysis, the presence of aDA at week 26 increased the risk of adalimumab discontinuation by 1.78 [IC 95%=1.11-2.85], p=0.016. Conclusion: Immunogenicity is a key factor that contributes to adalimumab discontinuation in SpA. MTX at initiation may therefore be considered in combination to adalimumab in SpA patients. References [1] Ducourau E, et al. Methotrexate reduces adalimumab immunogenicity in patients with spondyloarthritis: a randomized clinical trial. EULAR17-1527. Acknowledgement: This work was promoted by the Regional University Hospital Center of Tours and supported by grants from the French Ministry for Health and Sport within the framework of the Programme Hospitalier de Recherche Clinique 2011. Disclosure of interests: Marine Samain: None declared, Emilie Ducourau Speakers bureau: BMS and abbvie, theo Rispens Grant/research support from: Genmab, Speakers bureau: Pfizer, abbvie, Regeneron, Emmanuelle Dernis: None declared, Fabienne Le Guilchard: None declared, Lucia andras: None declared, aleth Perdriger: None declared, Eric Lespessailles Grant/research support from: Grants/research support from amgen, Eli Lily, MSD, UCB., Consultant for: Consultant for amgen, Expanscience, Eli Lilly, MSD, UCB., antoine Martin: None declared, Gregoire Cormier: None declared, Thomas armingeat: None declared, Valerie Devauchelle-Pensec Grant/research support from: Roche-Chugai, Speakers bureau: MSD, BMS, UCB, Roche, Elisabeth Gervais Speakers bureau: abbvie, BMS, MSD, Pfizer, Roche, UCB, Novartis, Benoit Le Goff Speakers bureau: abbvie, BMS, Janssen, MSD, Pfizer, Sanofi-Genzyme, UCB, Novartis, annick de Vries: None declared, Eric Piver: None declared, Gilles Paintaud Grant/research support from: Novartis, Roche Pharma, Sanofi-Genzyme, Chugai, Pfizer and Shire, Celine Desvignes: None declared, David Ternant Speakers bureau: Sanofi, amgen, Herve Watier: None declared, Philippe Goupille Grant/research support from: Financial compensation received from MSD on a pro-rota basis for participation in Scientific Committee meetings and functions for this study, Speakers bureau: abbvie, Biogaran, BMS, Hospira, Janssen, MSD, Pfizer, Sanofi-Genzyme, UCB, Denis Mulleman Speakers bureau: Pfizer, Novartis, Grifols

Introduction La spondyloarthrite (SpA) est un rhumatisme inflammatoire chronique touchant 0,1 a 1,4 % de la population. A ce jour, il n’existe pas de marqueur biologique assez specifique et sensible pour evaluer l’activite de la SpA et donc pour predire la reponse therapeutique precocement. L’objectif de notre etude etait d’analyser l’evolution de la concentration serique de calprotectine apres l’initiation de l’adalimumab dans la SpA axiale et sa relation avec l’activite de la maladie. Patients et methodes Trente patients presentant une SpA qui ont participe a l’essai clinique CoMARIS ( NCT01895764 ) ont ete analyses. Tous les patients ont recu 40 mg d’adalimumab par voie sous-cutanee (SC) toutes les deux semaines, soit seuls (n = 12), soit en association avec du methotrexate (n = 18). La concentration serique de calprotectine a ete mesuree 2 semaines avant, 4 et 12 semaines apres l’initiation de l’adalimumab. Le score d’activite de la maladie de la spondylarthrite ankylosante (ASDAS) et le C-reactive protein (CRP) etaient disponibles. L’indice de correlation de Spearman a ete mesure a chaque point dans le temps entre l’ASDAS et la calprotectine. Ensuite, la variation de la calprotectine (mediane [intervalle]) selon les categories de reponse ASDAS (« aucune amelioration », n = 15 ; « amelioration importante », n = 7 ; « amelioration majeure », n = 5) a la semaine 12 a ete analysee. Resultats L’indice de correlation de Spearman entre la calprotectine et l’ASDAS etait de 0,29, 0,15 et 0,04 a la semaine −2, la semaine 4 et la semaine 12, respectivement. La diminution de la calprotectine semblait etre plus prononcee chez les patients ayant un ASDAS plus eleve au depart que chez ceux avec des valeurs d’ASDAS faibles. La variation de calprotectine entre la semaine 2 et la semaine 4 etait de −31,0 % [−43,7 ; −9,3] dans le « groupe d’amelioration majeure », −28,1 [−28,7 ; 17,2] dans le « groupe d’amelioration importante » et −27,9 [−45,7 ; −1,0]. La calprotectine diminuait de maniere moins prononcee par rapport a la CRP apres l’initiation de l’adalimumab. Conclusion La calprotectine est faiblement correlee a l’activite de la maladie dans la SpA axiale et est moins sensible au traitement par l’adalimumab, que la CRP.

Introduction Mechanisms of maintenance of both atrial fibrillation and structural left ventricular disease are known to include fibrosis. Galectin-3, a biomarker of fibrosis, is elevated both in patients with heart failure and persistent atrial fibrillation. We sought to find whether galectin-3 has a prognostic value in patients with heart failure and a reduced left ventricular ejection fraction undergoing ablation of persistent atrial fibrillation. Methods Serum concentrations of galectin-3 were determined in a consecutive series of patients with an ejection fraction ≤40%, addressed for ablation of persistent atrial fibrillation. Responders to ablation were patients in sinus rhythm and with an ejection fraction ≥50% at 6 months. A combined endpoint of heart failure hospitalization, transplantation and/or death was used at 12 months. Results Seventy-five patients were included (81% male, age 63±10 years, ejection fraction 34±7%, galectin-3 21±12 ng/mL). During follow-up, eight patients were hospitalized for decompensated heart failure, 1 underwent heart transplantation, and 4 died; 50 patients were considered as responders to ablation. After adjustment, galectin-3 level independently predicted both 6-month absence of response to ablation (OR = 0.89 per unit increase, p = 0.002). Patients with galectin-3 levels

Maintenance of atrial fibrillation is a complex mechanism, including extensive electrical and structural remodeling of the atria which involves progressive fibrogenesis. Galectin-3 is a biomarker of fibrosis, and, thus, may be involved in atrial remodeling in atrial fibrillation patients. We review the role of galectin-3 in AF mechanisms and its potential therapeutic implications.

Background TNF inhibitors are effective in treating spondyloarthritis (SpA). However, anti-drug antibodies (ADA) may be responsible for decreased efficacy. Methotrexate reduces adalimumab immunogenicity in rheumatoid arthritis (1). Objectives The aim of the study was to evaluate the effect of methotrexate on ADA detection in SpA patients receiving adalimumab. Methods One hundred and ten SpA patients eligible for adalimumab 40 mg S/C eow were randomized on a 1:1 ratio to receive MTX 10 mg s/c every week, 2 weeks prior (V0) adalimumab (MTX+), or adalimumab alone (MTX-). ADA detection and adalimumab concentration were assessed 4 weeks (V2), 8 weeks (V3), 12 weeks (V4) and 26 weeks (V5) after starting adalimumab (V1). The main outcome was the percentage of positive patients for ADA detection at V5 or last available visit. Results Patients9 characteristics (sex, previous TNF inhibitors, disease duration, HLA B27 +/-, BMI) were comparable between the two groups. One hundred and seven patients were analyzed, 55 in the MTX+ group versus 52 in the MTX- group. ADA were detected at V5, in 39/107 (36.4%) patients; 13/52 (25%) in the MTX+ group versus 26/55 (47.3%) in the MTX- group (p=0.03). Adalimumab concentrations were statistically higher in the MTX+ group as compared with the MTX- group at V2, V3, V4 and V5 (Figure 1). There was no difference in terms of adverse events and efficacy between the two groups. Conclusions MTX reduces immunogenicity and ameliorates pharmacokinetics of adalimumab in SpA patients. The clinical impact of this combination requires longer period studies. References Krieckaert CL, Nurmohamed MT, Wolbink GJ. Ann Rheum Dis 2012. Acknowledgements The authors thank Bruno Giraudeau for his methodological advice in the study design; Yoann Desvignes for technical support with the study protocol; Elody Marnat for data management; Coraline Gadras and Celine Vignault for blood sample management; Anne-Claire Duveau for technical assistance. This work was promoted by the Regional University Hospital Center of Tours and supported by grants from the French Ministry for Health and Sport within the framework of the “Programme Hospitalier de Recherche Clinique 2011” and received fundings from the “Lions Club, Tours Val de France” and from Nordic Pharma who provided subcutaneous methotrexate. This work was a collaborative venture by HUGO (Hopitaux Universitaires du Grand Ouest – Western France University Hospitals Network). Disclosure of Interest E. Ducourau: None declared, T. Rispens: None declared, E. Dernis: None declared, F. Le Guilchard: None declared, L. Andras: None declared, A. Perdriger: None declared, E. Lespessailles Grant/research support from: MSD, Novartis, UCB, Pfizer, Consultant for: MSD, Novartis, UCB, Pfizer, A. Martin: None declared, G. Cormier: None declared, T. Armingeat: None declared, V. Devauchelle-Pensec: None declared, E. Solau-Gervais: None declared, B. Le Goff: None declared, A. de Vries: None declared, E. Piver: None declared, D. Ternant Consultant for: Sanofi, Amgen, P. Goupille Consultant for: Abbvie, Biogaran, BMS, Hospira, Janssen, MSD, Pfizer, Sanofi-Genzyme, UCB, D. Mulleman Grant/research support from: Nordic Pharma, Abbvie, Consultant for: MSD, Novartis, UCB, Pfizer

Converging evidence highlights that lipid metabolism plays a key role in ALS pathophysiology. Dyslipidemia has been described in ALS patients and may be protective but peripheral lipoprotein subclasses have never been studied.We collected sera from 30 ALS patients and 30 gender and age-matched controls. We analyzed 11 distinct lipoprotein subclasses by linear polyacrylamide gel electrophoresis (Lipoprint, Quantimetrix Corporation, USA). We also measured lipoprotein (a), apolipoprotein B, and apolipoprotein E levels.ALS patients had significant higher total cholesterol, HDL-cholesterol, and LDL-cholesterol levels than controls (p0.0001, p=0.0007, and p=0.0065, respectively). The LDL-1 subfraction concentration was higher (1.03±0.41 vs. 0.71±0.28mmol/L; p=0.0006) and the IDL-B subfraction lower (6.5±2% vs. 8.0±2%; p=0.001) in ALS patients than controls.Our preliminary work confirmed the association between ALS and dyslipidemia. The low IDL-B levels may explain the hepatic steatosis frequently reported in ALS. The high levels of the cholesterol-rich LDL-1 subfraction is consistent with previously reported hypercholesterolemia.This study describes, for the first time, the distribution of serum lipoproteins in ALS patients, with low IDL-B and high LDL-1 subfraction level.

International audience; OBJECTIVE: The study aims to evaluate the accuracy of combining uterine artery Doppler (UAD), PlGF and sFlt-1 in the first trimester for preeclampsia screening. METHODS: Prospectively enrolled women at high risk of preeclampsia were included. Transabdominal UAD measurements and serum biomarkers were collected between 11 and 13 weeks of gestation in three university hospitals and in one general hospital. The main outcome was preeclampsia. UAD parameters and biomarker levels among women with preeclampsia were compared with those of women in the unaffected group in univariate and multivariate analyses. RESULTS: Out of 226 women included from May 2007 to January 2011, 27 (11.9%) women developed preeclampsia. Among women affected by preeclampsia, the lowest pulsatility index was higher (p\,=\,0.02), bilateral notching was more frequent (p\,=\,0.01), and PlGF was lower (p\

BackgroundMechanisms of maintenance of atrial fibrillation are known to include fibrosis. Galectin-3, as a biomarker of fibrosis, may be a valuable marker of atrial remodeling. We sought to find whether there was a link between clinical features and higher galectin-3 levels in patients with atrial fibrillation.MethodsSerum concentrations of Galectin-3 were determined in a consecutive series of patients addressed for ablation of atrial fibrillation.ResultsOne-hundred-and-eighty-seven patients were included, 56% having a paroxysmal type of atrial fibrillation. Mean Galectin-3 concentration was 14.5 ± 5.5 ng/mL. Age, persistent form of atrial fibrillation, underlying cardiac disease, heart failure, decreased left ventricular ejection fraction (LVEF), hypertension, diabetes, treatment with ACEI/ARB, enlarged left atrium and renal insufficiency were associated with higher Galectin-3 levels. Importantly, persistent form of atrial fibrillation, female sex, and LVEF < 45% were independent predictors (OR 13.9, p = 0.01, OR = 11.7, p = 0.03, and OR 54.2, p = 0.04, respectively) of higher Galectin-3 levels (≥ 15 ng/mL).ConclusionsPersistent type of atrial fibrillation is an independent predictor of higher Galectin-3 concentration. This biomarker of fibrosis may be implied in the mechanisms of atrial remodeling and maintenance of atrial fibrillation, and thus be helpful for the design of therapeutic strategy in patients with atrial fibrillation.

The development of metastases largely relies on the capacity of cancer cells to invade extracellular matrices (ECM) using two invasion modes termed ‘mesenchymal' and ‘amoeboid', with possible transitions between these modes. Here we show that the SCN4B gene, encoding for the β4 protein, initially characterized as an auxiliary subunit of voltage-gated sodium channels (NaV) in excitable tissues, is expressed in normal epithelial cells and that reduced β4 protein levels in breast cancer biopsies correlate with high-grade primary and metastatic tumours. In cancer cells, reducing β4 expression increases RhoA activity, potentiates cell migration and invasiveness, primary tumour growth and metastatic spreading, by promoting the acquisition of an amoeboid–mesenchymal hybrid phenotype. This hyperactivated migration is independent of NaV and is prevented by overexpression of the intracellular C-terminus of β4. Conversely, SCN4B overexpression reduces cancer cell invasiveness and tumour progression, indicating that SCN4B/β4 represents a metastasis-suppressor gene., The capacity of cancer cells to migrate is intimately linked to their ability to induce metastasis. Here the authors show that the sodium channel β4 subunit regulates breast cancer cell migration via inhibition of RhoA activation, independently from its function as an auxiliary protein of the sodium channel.

Objective This study aimed to determine if screening for preeclampsia could be improved between 20 and 24 weeks of gestation by uterine artery Doppler (UAD), biomarkers and lipid-related markers. Method Women at high risk of preeclampsia according to obstetric and medical characteristics and history were prospectively enrolled. Transabdominal UAD, serum biomarkers (placenta growth factor (PlGF), soluble Fms-like tyrosine kinase-1 and sEndoglin) and lipid-related markers (total and high-density lipoprotein cholesterol, triglycerides and leptin) were prospectively collected between 20 and 24 weeks of gestation. The main endpoint was preeclampsia. UAD indices and biomarker levels were compared for the groups with and without preeclampsia. Multivariate analyses took into account the laboratory and ultrasound variables significantly associated with preeclampsia in the univariate analysis and age and nulliparity. Results The study at Tours University Hospital took place from March 2003 to February 2008 and included 235 women: 56 (23.8%) developed preeclampsia, 42 were severe (17.8%) and 14 occurred before 34 weeks of gestation (5.9%). The group with preeclampsia had a higher UAD pulsatility index (p = 0.0003), more frequent bilateral notches (p = .0001), lower PlGF levels (239.90 vs 302.00 pg/mL; p = 0.0015), and higher triglyceride (1.95 vs 1.70 mmol/l; p = 0.0068) and leptin levels (37.40 vs 22.55 ng/mL; p = < 0.001). No significant differences were observed for other markers. Screening for preeclampsia using these Doppler and laboratory findings produced an area under the curve of 0.795. Conclusion Second-trimester UAD findings, and PlGF, triglyceride and leptin levels could help to predict preeclampsia. © 2013 John Wiley & Sons, Ltd.

Hepatocellular carcinoma (HCC) is a common outcome of chronic hepatitis C virus (HCV) infection and constitutes the main burden of this disease. The molecular mechanisms underlying the development of HCC are multiple and might involve certain microRNA (miR). As discordant results have been reported concerning the detection of expression of miR-152 and miR-122 in HCC, our aim was to measure the levels of both miRs in serum and liver samples.We analyzed miR-152 and miR-122 expression by reverse transcription-quantitative polymerase chain reaction in a serum cohort from 14 HCV-infected patients who developed HCC, 20 HCV+ patients without HCC, and 19 control patients. We also studied miR-152 and miR-122 in an independent tissue cohort from 11 normal livers, and from paired HCC and non-tumor adjacent livers of 11 HCV-infected patients and 12 non-infected patients.In serum samples, higher levels of miR-122 were found in non-HCC HCV+ compared to HCC HCV+ and control groups, whereas miR-152 was detectable in a lower range in HCC HCV+ compared to non-HCC HCV+ and control groups. We found higher signals for miR-122 and miR-152 in non-tumor liver and HCC tissues compared to control tissues. Hepatocellular carcinoma etiology had no detectable influence on miR-122 expression, whereas miR-152 was increased in HCV+ tissue samples.Detection of low values of circulating miR-152 is a potentially interesting marker of hepatocarcinogenesis in HCV+ patients, in contrast to miR-122, which varies according to hepatocyte damage.

Galectin-3 is a biomarker of fibrosis and atrial remodeling, involved in the mechanisms of initiation and maintenance of atrial fibrillation (AF). We sought to study the accuracy of galectin-3 level in predicting recurrences of AF after ablation. Serum concentrations of galectin-3 were determined in a consecutive series of patients addressed for AF ablation in our center. After a 3-month blanking period, recurrences of atrial arrhythmias were collected during the first year in all patients, using Holter monitoring at 3, 6 months and 12 months. A total of 160 patients were included, with a mean galectin-3 rate was 14.4 ± 5.6 ng/mL. At 12-month, 55 patients (34%) had reexperienced sustained atrial arrhythmia. Only higher galectin-3 level (HR = 1.07 [1.01–1.12], p = 0.02) and larger left atrial diameter (HR = 1.07 [1.03–1.12], p = 0.001) independently predicted recurrence. Patients with both galectin-3 level

Objective HCV particles are associated with very low-density lipoprotein components in chronically infected patients. These hybrid particles, or ‘lipo-viro particles’ (LVPs), are rich in triglycerides, and contain the viral RNA, the capsid protein, E1E2 envelope glycoproteins and apolipoproteins B and E. However, their specific ultrastructural organisation has yet to be determined. We developed a strategy for the preparation of any viral sample that preserves the native structure of the LVPs, facilitating their precise morphological characterisation. Design Using a strategy based on the direct specific immunocapture of particles on transmission electron microscopy (TEM) grids, we characterised the precise morphology of the viral particle by TEM. Results The LVP consists of a broad nucleocapsid surrounding an electron-dense centre, presumably containing the HCV genome. The nucleocapsid is surrounded by an irregular, detergent-sensitive crescent probably composed of lipids. Lipid content may determine particle size. These particles carry HCV E1E2, ApoB and ApoE, as shown in our immuno-EM analysis. Our results also suggest that these putative LVPs circulate in the serum of patients as part of a mixed population, including lipoprotein-like particles and complete viral particles. Conclusions Twenty-five years after the discovery of HCV, this study finally provides information about the precise morphological organisation of viral particles. It is truly remarkable that our TEM images fully confirm the ultrastructure of LVPs predicted by several authors, almost exclusively from the results of molecular biology studies.

Background: Multiple myeloma (MM) is characterized by the accumulation of malignant plasma cells, which secrete >30g/L of monoclonal immunoglobulin (mc Ig). MM is always preceded by the usually asymptomatic stage called "monoclonal gammapathy of undetermined significance" (MGUS). Presently the detection of a mc Ig does not lead to analysis of its specificity of antigen (Ag) recognition. Yet certain infectious pathogens are associated with the development of B-cell malignancy: for instance, Epstein-Barr virus (EBV) and Burkitt lymphoma, Helicobacter pylori (H. pylori) and MALT lymphoma. Objective: Our aim was to determine the frequency of recognition by purified mc Ig from MGUS and MM patients, of infectious pathogens associated with chronic infection and B-cell malignancy. Methods: We used the "Multiplex Infectious Antigen Array" (MIAA)test set up by the team to study the specificity of purified mc IgG against 8 infectious pathogens: hepatitis C virus (HCV), EBV, H. pylori, varicella zoster virus (VZV), cytomegalovirus (CMV), herpesvirus simplex 1 (HSV-1), HSV-2, and Toxoplasma gondii. Second or third methods were used to verify the specificity of mc IgG (western blotting, ELISA, or another multiplex array). Sialylation of mc Ig, which reflects chronic inflammation, was also studied, via ELISAs and Western blotting, for 98 MGUS and MM patients and 43 controls. Results: The specificity of purified mc IgG was analyzed for 291 patients (60 MGUS, including 4 associated with a B-cell lymphoma; 108 MM; for the 123 other patients the diagnosis of MGUS or MM was not available). Purified mc IgG were found to be specific for one of the 8 infectious pathogens of the MIAA test for 17.5% of patients (51/291: 25 MGUS, 1 SMM, and 25 MM). Purified mc Ig specifically recognized EBV (n=27, including 8 MGUS, 1 SMM and 18 MM ; the Ag recognized were EBNA-1, 25 cases; and VCA, 2 cases); HCV (10 cases: 9 MGUS and 1 MM) ; HSV-1 (7 cases: 5 MGUS including 3 associated with a B-cell malignancy, and 2 MM) ; H. pylori (5 cases: 2 MGUS including 1 associated with a B-cell malignancy, and 3 MM); VZV (1 MM); and CMV (1 MGUS). The Ag most frequently recognized by purified mc IgG was EBV EBNA-1; MM patients with EBNA-1-specific mc IgG represented 10% of the MM in this series. Consistent with a pro-inflammatory effect, for 81.6% of MGUS and MM patients (80/98), the purified mc IgG was found to be hyposialylated. Moreover, purified polyclonal Ig were also hyposialylated for 37% of MM (compared to Conclusion: At least 6 infectious pathogens (EBV, HCV, H. pylori, HSV, VZV, CMV) may be involved in the pathogenesis of MGUS and MM with mc IgG. Importantly, EBV, HCV, H. pylori and HSV are known as oncogenic pathogens associated with solid cancer and/or B-cell malignancy. Finally, for 37% of the MM patients examined, both the polyclonal and mc Ig were found to be hyposialylated, which suggests that inflammation existed prior to clonality and MM. Disclosures Garderet: Takeda: Consultancy; Amgen: Consultancy; BMS: Consultancy, Honoraria; Novartis: Consultancy.

The advent of infectious molecular clones of Hepatitis C virus (HCV) has unlocked the understanding of HCV life cycle. However, packaging of the genomic RNA, which is crucial to generate infectious viral particles, remains poorly understood. Molecular interactions of the domain 1 (D1) of HCV Core protein and HCV RNA have been described in vitro. Since compaction of genetic information within HCV genome has hampered conventional mutational approach to study packaging in vivo, we developed a novel heterologous system to evaluate the interactions between HCV RNA and CoreD1. For this, we took advantage of the recruitment of Vpr fusion-proteins into HIV-1 particles. By fusing HCV Core D1 to Vpr we were able to package and transfer a HCV subgenomic replicon into a HIV-1 based lentiviral vector. We next examined how deletion mutants of basic sub-domains of Core D1 influenced HCV RNA recruitment. The results emphasized the crucial role of the first and third basic regions of D1 in packaging. Interestingly, the system described here allowed us to mobilise full-length JFH1 genome in CD81 defective cells, which are normally refractory to HCV infection. This finding paves the way to an evaluation of the replication capability of HCV in various cell types.

Hepatitis C virus (HCV) is a major cause of chronic hepatitis associated with liver steatosis, commonly evolving to cirrhosis or hepatocellular carcinoma. The World Health Organisation (WHO) estimates that there are around 170 million chronic HCV carriers worldwide. The virus has a highly variable sequence, allowing definition of seven genotypes with different geographical distributions. Both clinical outcome and response to antiviral therapy are strongly influenced by HCV genotype. Importantly, several recent papers have suggested that the lipid profile of infected patients is strongly indicative of the various clinical outcomes of HCV infection. Furthermore, viral molecular and cellular studies have shown a tight link between cellular lipid metabolism and almost every step of the HCV infectious cycle. In the present review we summarise the current knowledge establishing the interplay between the molecular features of HCV replication, the cellular lipid biology and the lipid profiles observed in the serum of infected patients.

International audience; Procalcitonin (PCT) levels are commonly used for diagnostic guidance in routine bacterial infections. By contrast, little data are currently available regarding PCT in parasitic diseases, and its role in cases of invasive amoebiasis has not yet been described. For this purpose, 35 adult patients with a proven diagnosis of invasive or digestive amoebiasis were included in a 4-year study period. Serum PCT was retrospectively assessed. Results were analysed with regard to the usual inflammatory biomarkers, like C-reactive protein (CRP). PCT was significantly higher in patients with proven invasive amoebiasis than in digestive amoebiasis (mean value: 4.03 μg/L versus 0.07 μg/L, respectively; P < 0.001), but the SD was greater than with CRP, and the effect was less than that demonstrated in bacterial infections. By contrast, PCT was not shown to be elevated during digestive amoebiasis.

This study aims to develop a cellular metabolomics model that reproduces the pathophysiological conditions found in amyotrophic lateral sclerosis in order to improve knowledge of disease physiology. We used a co-culture model combining the motor neuron-like cell line NSC-34 and the astrocyte clone C8-D1A, with each over-expressing wild-type or G93C mutant human SOD1, to examine amyotrophic lateral sclerosis (ALS) physiology. We focused on the effects of mutant human SOD1 as well as oxidative stress induced by menadione on intracellular metabolism using a metabolomics approach through gas chromatography coupled with mass spectrometry (GC-MS) analysis. Preliminary non-supervised analysis by Principal Component Analysis (PCA) revealed that cell type, genetic environment, and time of culture influenced the metabolomics profiles. Supervised analysis using orthogonal partial least squares discriminant analysis (OPLS-DA) on data from intracellular metabolomics profiles of SOD1G93C co-cultures produced metabolites involved in glutamate metabolism and the tricarboxylic acid cycle (TCA) cycle. This study revealed the feasibility of using a metabolomics approach in a cellular model of ALS. We identified potential disruption of the TCA cycle and glutamate metabolism under oxidative stress, which is consistent with prior research in the disease. Analysis of metabolic alterations in an in vitro model is a novel approach to investigation of disease physiology.