Search

Systemic sclerosis is a rare autoimmune disease associated with rapidly evolving interstitial lung disease, responsible for the disease severity and mortality. Specific biomarkers enabling the early diagnosis and prognosis associated with the disease progression are highly needed. Volatile organic compounds in exhaled breath are widely available and non-invasive and have the potential to reflect metabolic processes occurring within the body. Comprehensive two-dimensional gas chromatography coupled to high-resolution mass spectrometry was used to investigate the potential of exhaled breath to diagnose systemic sclerosis. The exhaled breath of 32 patients and 30 healthy subjects was analyzed. The high resolving power of this approach enabled the detection of 356 compounds in the breath of systemic sclerosis patients, which was characterized by an increase of mainly terpenoids and hydrocarbons. In addition, the use of 4 complementary statistical approaches (two-tailed equal variance t-test, fold change, partial least squares discriminant analysis, and random forest) resulted in the identification of 16 compounds that can be used to discriminate systemic sclerosis patients from healthy subjects. Receiver operating curves were generated that provided an accuracy of 90%, a sensitivity of 92%, and a specificity of 89%. The chemical identification of eight compounds predictive of systemic sclerosis was validated using commercially available standards. The analytical variations together with the volatile composition of room air were carefully monitored during the timeframe of the study to ensure the robustness of the technique. This study represents the first reported evaluation of exhaled breath analysis for systemic sclerosis diagnosis and provides surrogate markers for such disease.

SSc is a rare disease of unknown origin associated with multiple organ involvement. One of the major complications that drives the mortality of SSc patients is interstitial lung disease. The course of SSc-interstitial lung disease progression has a wide spectrum. Since the treatment is based on aggressive immunosuppression it should not be given to stable or non-progressing disease. The correct identification of disease with high risk of progression remains a challenge for early therapeutic intervention, and biomarkers remain urgently needed. In fact, eight categories of biomarkers have been identified and classified according to the different biological pathways involved. The purpose of this article is to describe the main biomarkers thought to be of interest with clinical value in the diagnosis and prognosis of SSc-interstitial lung disease.

Background: Systemic sclerosis (SSc) is a rare connective tissue disease associated with rapid evolving interstitial lung disease (ILD), driving its mortality. Specific biomarkers associated with the progression of this lung disease are highly needed. We aimed to identify specific biomarkers of SSc-ILD to predict the evolution of the disease.Methods: We compared prospectively serum levels of several biomarkers associated with lung fibrosis in SSc patients (n=102), among which SSc-no ILD (n=63) and SSc-ILD (n=39), compared to healthy subjects (HS) (n=39). We also performed a longitudinal study in a subgroup of 28 patients analyzing biomarkers variations and pulmonary function tests over a period of 2 years.Results: Serum level of IGFBP-2 was significantly increased in SSc patients compared to HS, and negatively correlated with pulmonary function (assessed by carbon monoxide transfer coefficient (KCO)) (r=-0.29, p 105 ng/ml was associated with a poor outcome (KCO Conclusions: We showed for the first time that serum levels of IGFBP-2 might be a prognostic factor of the development of SSc-ILD. Indeed, initial level of IGFBP-2 above 105 ng/ml was associated with a poor patient’s outcome (KCO

Systemic sclerosis (SSc) is a potentially serious and disabling connective tissue disease specially in case of interstitial lung disease (SSc-ILD). The aim of our study was to evaluate the potential utility of dosing in the induced sputum (IS) and to compare their levels in SSc-ILD and SSc-nonILD patients, as well as in healthy volunteers (HV). IS and sera values were also compared. In a prospective cross-sectional analysis, we studied the IS and serum provided from 25 SSc patients, 15 SSc-nonILD and 10 SSc-ILD, compared to 25 HV. We analyzed sputum cell composition and quantified in the supernatant and corresponding serum by commercially available immunoassays: IGFBP-1, IGFBP-2, IGFBP-3, TGF-β, IL-8, TNF-α, YKL-40, MMP-7 and MMP-9. Lung function was studied by the determination of FEV-1 (%), FVC (%), DLCO (%) and KCO (%). The IS of SSc patients had a lower weight than HV (p

BackgroundSystemic sclerosis (SSc) is a potentially serious and disabling connective tissue disease specially in case of interstitial lung disease (SSc-ILD). The aim of our study was to evaluate the potential utility of dosing in the induced sputum (IS) and to compare their levels in SSc-ILD and SSc-nonILD patients, as well as in healthy volunteers (HV). IS and sera values were also compared.MethodsIn a prospective cross-sectional analysis, we studied the IS and serum provided from 25 SSc patients, 15 SSc-nonILD and 10 SSc-ILD, compared to 25 HV. We analyzed sputum cell composition and quantified in the supernatant and corresponding serum by commercially available immunoassays: IGFBP-1, IGFBP-2, IGFBP-3, TGF-β, IL-8, TNF-α, YKL-40, MMP-7 and MMP-9. Lung function was studied by the determination of FEV-1 (%), FVC (%), DLCO (%) and KCO (%).ResultsThe IS of SSc patients had a lower weight than HV (pConclusionOur study showed that SSc patients exhibit raised IS levels of IGFBP-1, TGF-β, IL-8, YKL-40 and MMP-7, molecules known to be involved in lung remodeling and fibrotic process, without any significant difference between SSc-ILD and SSc-nonILD patients. IL-8, TGF-□ and IGFBP-2 are correlated with lung function in SSc patients which emphasize clinical relevance. IS analysis represents a new approach to understand lung inflammatory process in SSc patients. A longitudinal study is needed to evaluate their pathophysiological relevance.

BackgroundSystemic sclerosis (SSc) is a rare connective tissue disease associated with rapid evolving interstitial lung disease (SSc-ILD), driving its mortality. Specific biomarkers associated with the evolution of the lung disease are highly needed. We aimed to identify specific biomarkers of SSc-ILD to predict the evolution of the disease. Nucleosomes are stable DNA/protein complexes that are shed into the blood stream making them ideal candidates for biomarkers.MethodsWe studied circulating cell-free nucleosomes (cf-nucleosomes) in SSc patients, 31 with ILD (SSc-ILD) and 67 without ILD. We analyzed plasma levels for cf-nucleosomes and investigated whether global circulating nucleosome levels in association with or without other biomarkers of interest for systemic sclerosis or lung fibrosis (e.g., serum growth factors: IGFBP-1 and the MMP enzyme: MMP-9), could be suitable potential biomarkers for the correct identification of SSc-ILD disease.ResultsWe found that H3.1 nucleosome levels were significantly higher in patients with SSc-ILD compared SSc patients without ILD (p< 0.05) and levels of MMP-9 were significantly increased in patients with SSc-ILD compared to SSc patients without ILD (p< 0.05). Conversely, IGFBP-1 was significantly reduced in patients with SSc-ILD compared to SSc without ILD (p< 0.001). The combination of cf-nucleosomes H3.1 coupled to MMP-9 and IGFBP-1 increased the sensitivity for the differential detection of SSc-ILD. High levels of accuracy were reached with this combined model: its performances are strong with 68.4% of positive predictive value and 77.2% of negative predictive value for 90% of specificity. With our model, we identified a significant negative correlation with FVC % pred (r= −0.22) and TLC % pred (r= −0.31). The value of our model at T1 (baseline) has a predictive power over the Rodnan score at T2 (after 6-18 months), showed by a significant linear regression withR2= 19% (p= 0.013). We identified in the sole group of SSc-ILD patients a significant linear regression with aR2= 54.4% with the variation of DLCO between T1 and T2 (p< 0.05).ConclusionIn our study, we identified a new blood-based model with nucleosomic biomarker in order to diagnose SSc-ILD in a SSc cohort. This model is correlated with TLC and FVC at baseline and predictive of the skin evolution and the DLCO. Further longitudinal exploration studies should be performed in order to evaluate the potential of such diagnostic and predictive model.

We read with interest the article of Bozzalla Cassione et al about COVID-19 incidence in their systemic lupus erythematosus (SLE) cohort.1 Their study adds useful epidemiological information about COVID-19 risk in SLE.1 They suggest that hydroxycholoroquine was not protective, but could not draw definite conclusion and open the question to immunosuppressive drugs’ influence. We would like to share analysis of our SLE cohort (n=225) that can help to answer these questions and determine COVID-19 infection risk factors. Determining COVID-19 incidence is challenging: PCR lacks sensitivity, was usually realised only in severely ill patients and patients with suggestive benign symptoms could stay at home without medical contact. We studied the incidence of COVID-19 infection, either asserted or suspected, by analysing positive nasopharyngeal PCR, hospitalisation or contact with emergency department, but also suspected diagnosis in ambulatory medicine. Each patient was called by phone to determine COVID-19 suggestive symptoms since 4 February 2020, date of the first case in our country. Among our patients, 92.9% were female, with a mean (±SD) age of 51.7 (±14.9) years. Most recent biological evaluation showed positivity for ds-DNA in 24% (median (min–max) levels: 139 (12–758) IU/mL). Mean (±SD) number of 1997 American College of Rheumatology (ACR), 2019 American College of Rheumatology/European League Against Rheumatism (ACR/EULAR) and Systemic Lupus International Collaborating Clinics (SLICC) classification criteria were 4.5 (±1.5), …

Background: Systemic sclerosis (SSc) is a rare connective tissue disease associated with potential rapid evolving interstitial lung disease (SSc-ILD), driving the mortality of these patients. Therefore, a specific biomarker associated with the evolution of that disease is highly needed to identify patients with an increased risk of death. Aim of the Study: Identify specific biomarkers of SSc-ILD to predict the evolution of the disease. Methods: In this prospective longitudinal study, we compared serum levels of biomarkers assumed to be associated with lung fibrosis (TGF-β, IGF-1, IGFBP-1, IGFBP-2, IGFBP-3, IL-8, MMP-7, MMP-9, YKL-40 and TNF-α) among three groups: SSc-ILD (n=39), SSc without ILD (n=63) and healthy subjects (HS)(n=39). We also prospectively analyzed variations of biomarkers and correlated them to pulmonary function tests (n=28). Then, we realized an in vitro analysis to study the potential anti-fibrotic effect of IGFBP-2 (n=3). Results: IGFBP-2, IL-8 and MMP-9 are increased in SSc patients compared to HS (p Conclusion: IGFBP-1 has a potential interest to identify early SSc-ILD whereas IGFBP-2 would rather predict the risk of a rapid evolution of lung fibrosis. Moreover, in vitro studies confirmed the anti-fibrotic effect of IGFBP-2 underlying its potential interest in further mechanical studies and therapeutic aspects.

Background The OMERACT Ultrasound scoring system (USSS) of joint in RA has been shown to be reliable and sensitive to change when used in clinical trial setting. However it is unclear whether this reliability is also achieved in clinical daily practice among Rheumatologists performing US in a non-research setting and using different machines. Objectives To assess agreement between non-research sonographers in scoring synovitis using the OMERACT USSS and using the scoring of an OMERACT expert as gold standard. To assess the reliability of the USSS by using different US machines. Methods First an OMERACT US expert presented the scoring method and supervised a training session. The wrist, MCP 2 and 3 joints of the left hand (dorsal aspect) of 3 RA patients were then successively evaluated by 9 Rheumatologists with 3 different US machines: Hitachi Arietta = H, GE Logic E9 = G, Esaote MyLab 7= E. The USSS included B-mode acquisition of synovial hypertrophy (SH), joint effusion (JE) and bone erosion (BE), and Power-Doppler (PD) activity. JE, SH and BE were scored binary; SH and PD were score semi quantitatively (0 to 3), by both the 9 participating Rheumatologists and the OMERACT US expert. The agreement between each participant with the scoring of the OMERACT US expert was quantified by proportions. The inter-US machine reproducibility was assessed by kappa statistics for discrete variables and weighted kappa9s for ordinate. Results The 3 joints of 3 patients each were evaluated on the 4 US items (SH, JF, BE, PD) on a different US machine. So 27 values were obtained for each joint within each item. The percentages of exact agreement (PEA) between these 27 values and the scores of the OMERACT US expert were calculated and ranged between 33% (9/27) for SH-MCP3, 37% for JF-MCP2 and PD-Wrist, 41% for SH-wrist, 48% BE-MCP3, 52% PD-MCP3, 56% PD-MCP3, 59 JF-wrist and BE-MCP2, 63% JF-MCP3, 67% SH-MCP2 and 78% (21/27) for BE-wrist. For the inter-US machines reproducibility, kappa between the 3 machines was calculated on 9 data (3 patients x 3 joints). The reliability was low for detecting JE ≤0.211. Acceptable reliability among machines was found for SH, BE and PD (table 1). Conclusions Non OMERACT-expert Rheumatologists can apply the USSS, and this score works well across different machines. The difference among machines and sonographers is mostly captured by the low reliability of JE. The OMERACT USSS could be used by sonographers in their everyday clinical practice to evaluate the activity of RA patients. Further studies in clinical-based settings will allow to define more precisely the applicability of this scoring system. Disclosure of Interest None declared

Background. - Early detection of pulmonary arterial hypertension (PH) is crucial in systemic scleroderma. However, predictors of new onset of resting PH during follow-up (FUPH) have been poorly explored. Aim. - To determine whether nailfold videocapillaroscopy (NVC) grade and exercise echocardiographic variables are predictors of FUPH. Methods. - We prospectively enrolled 40 patients with systemic sclerosis (age 54 13 years; 68% women). All patients underwent graded semisupine exercise echocardiography and NVC. Baseline resting PH and FUPH were defined as systolic pulmonary arterial pressure (sPAP) > 35 mmHg, and exercise-induced PH (EIPH) as exercise sPAP > 50 mmHg. Results. - Seventeen patients developed EIPH (43%). During follow-up (FU) (25 15 months), 11 patients without baseline PH developed FUPH (28%), all from the EIPH group. Patients with FUPH were significantly older (60 +/- 14 vs 50 +/- 12 years; P = 0.04), had higher resting and exercise sPAP (30 +/- 4 vs 22 +/- 5 and 60 +/- 12 vs 40 +/- 11 mmHg, respectively; P < 0.0001) and a higher exercise E/e' ratio (9.4 +/- 0.7 vs 5.8 +/- 0.4; P = 0.0003) and presented more frequently NVC grade > 2 (90% vs 35%; P = 0.0009). After adjustment for age, resting sPAP, exercise sPAP and NVC grade > 2 were associated with maximal resting sPAP during follow-up and FUPH (P < 0.05). Patients with both EIPH and NVC grade > 2 had a very high incidence of FUPH (82%), and both variables remained strongly associated with FUPH after adjustment for age (hazard ratio 11.6, 95% confidence interval 2.4-55.3; P = 0.002). Conclusion. - Exercise echocardiography and NVC can identify a subgroup of patients with systemic sclerosis who are at risk of developing FUPH.

The aim of this study was to assess rheumatoid arthritis (RA) synovitis with positron emission tomography (PET) and( 18)F-fluorodeoxyglucose ((18)F-FDG) in comparison with dynamic magnetic resonance imaging (MRI) and ultrasonography (US).Sixteen knees in 16 patients with active RA were assessed with PET, MRI and US at baseline and 4 weeks after initiation of anti-TNF-alpha treatment. All studies were performed within 4 days. Visual and semi-quantitative (standardised uptake value, SUV) analyses of the synovial uptake of FDG were performed. The dynamic enhancement rate and the static enhancement were measured after i.v. gadolinium injection and the synovial thickness was measured in the medial, lateral patellar and suprapatellar recesses by US. Serum levels of C-reactive protein (CRP) and metalloproteinase-3 (MMP-3) were also measured.PET was positive in 69% of knees while MRI and US were positive in 69% and 75%. Positivity on one imaging technique was strongly associated with positivity on the other two. PET-positive knees exhibited significantly higher SUVs, higher MRI parameters and greater synovial thickness compared with PET-negative knees, whereas serum CRP and MMP-3 levels were not significantly different. SUVs were significantly correlated with all MRI parameters, with synovial thickness and with serum CRP and MMP-3 levels at baseline. Changes in SUVs after 4 weeks were also correlated with changes in MRI parameters and in serum CRP and MMP-3 levels, but not with changes in synovial thickness.(18)F-FDG PET is a unique imaging technique for assessing the metabolic activity of synovitis. The PET findings are correlated with MRI and US assessments of the pannus in RA, as well as with the classical serum parameter of inflammation, CRP, and the synovium-derived parameter, serum MMP-3. Further studies are warranted to establish the place of metabolic imaging of synovitis in RA.

Objective To study the levels of matrix metalloproteinase-3 (MMP-3) in the knee synovial fluid (SF) of inflammatory arthropathies (rheumatoid arthritis whether erosive or not, reactive arthritis, acute crystal arthritis) and degenerative arthropathies [chronic crystal disease, osteoarthritis and (control) meniscus pathology] and to correlate them with the degree of joint destruction, local inflammatory and immune parameters and systemic markers of inflammation. Methods SF levels of MMP-3 (precursor, active and tissue inhibitor of MMP-bound forms), tumour necrosis factor (TNF) alpha, soluble TNF receptors I and II, interleukin (IL)-6 and soluble IL-6 receptor were measured by ELISA in 107 inflammatory and 53 degenerative arthropathies. Results MMP-3 levels in SF were (i) significantly higher in inflammatory than in degenerative arthropathies; (ii) not related to the degree of joint destruction; (iii) significantly correlated with the levels of all SF markers tested and with erythrocyte sedimentation rate and serum levels of C-reactive protein and fibrinogen. Conclusion Increased MMP-3 levels in SF are found in inflammatory arthropathies and are not specific for erosive joint diseases. MMP-3 in SF is therefore a potential candidate for the assessment of the inflammatory process in joints. However, the exclusive determination of the active form could indicate the degree of joint destruction.

SUMMARY Synovial fluid (SF) levels of soluble CD23 (sCD23) were determined in 96 patients presenting with an inflammatory knee effusion (73 with RA and 23 with reactive arthritis (ReA) serving as a control inflammatory non-erosive group) and were correlated with the degree of joint destruction, with local immune parameters (IL-1β, IL-3, IL-4, IL-6, IL-8, IL-10, IL-12 and sCD25) and with serum markers of inflammation, C-reactive protein and erythrocyte sedimentation rate. RA patients, classified as erosive or not according to Larsen’s grade, were separated as follows: (i) 13 patients with non-erosive RA; (ii) 16 RA patients with erosions in hands but not in knees, matched for disease duration with the first group; (iii) 44 RA patients with hand and knee erosions, matched with the second group for rheumatoid factor positivity but of longer disease duration. SF sCD23 levels were significantly increased in both erosive RA groups compared with non-erosive diseases, whether RA or ReA (P

Exercise-induced pulmonary arterial hypertension (EIPH) in systemic sclerosis (SSc) has already been observed but its determinants remain unclear. The aim of this study was to determine the incidence and the determinants of EIPH in SSc.We prospectively enrolled 63 patients with SSc (age 54±3years, 76% female) followed in CHU Sart-Tilman in Liège. All patients underwent graded semi-supine exercise echocardiography. Systolic pulmonary arterial pressure (sPAP) was derived from the peak velocity of the tricuspid regurgitation jet and adding the estimation of right atrial pressure, both at rest and during exercise. Resting pulmonary arterial hypertension (PH) was defined as sPAP35 mmHg and EIPH as sPAP50 mmHg during exercise. The following formulas were used: mean PAP (mPAP) = 0.61 × sPAP + 2, left atrial pressure (LAP)=1.9+1.24 × left ventricular (LV) E/e' and pulmonary vascular resistance (PVR)=(mPAP-LAP)/LV cardiac output (CO) and slope of mPAP-LVCO relationship=changes in mPAP/changes in LVCO. Resting PH was present in 3 patients (7%) and 21 patients developed EIPH (47%). Patients with EIPH had higher resting LAP (10.3 ± 2.2 versus 8.8 ± 2.3 mmHg; p = 0.03), resting PVR (2.6 ± 0.8 vs. 1.4 ± 1.1 Woods units; p=0.004), exercise LAP (13.3 ± 2.3 vs. 9 ± 1.7 mmHg; p0.0001), exercise PVR (3.6 ± 0.7 vs. 2.1 ± 0.9 Woods units; p = 0.02) and slope of mPAP-LVCO (5.8 ± 2.4 vs. 2.9 ± 2.1 mmHg/L/min; p0.0001). After adjustment for age and gender, exercise LAP (β=3.1 ± 0.8; p=0.001) and exercise PVR (β=7.9 ± 1.7; p=0.0001) were independent determinants of exercise sPAP.EIPH is frequent in SSc patients and is mainly related to both increased exercise LV filling pressure and exercise PVR.

Extracellular proteolysis is considered to be required during neuritic outgrowth to control the adhesiveness between the growing neurite membrane and extracellular matrix proteins. In this work, PC12 nerve cells were used to study the modulation of proteolytic activity during neuronal differentiation. PC12 cells were found to contain and release a 70–75-kDa tissue-type plasminogen activator (tPA) and a much less abundant 48-kDa urokinase-type plasminogen activator. A plasminogen activator inhibitor (PAI) activity with molecular sizes of 54 and 58 kDa was also detected in PC12 cell conditioned medium and formed high-molecular-mass complexes with released tPA. Release of PAI activity was dependent on treatment with nerve growth factor (NGF), whereas tPA synthesis and release were under control of a cyclic AMP- dependent mechanism and increased on treatment with dibutyryl-cyclic AMP [(But)2cAMP] or cholera toxin. Simultaneous treatment with NGF and (But)2cAMP resulted in increases of both tPA and PAI release and enhancement of tPA-PAI complex formation. The resulting plasminogen activator activity in conditioned medium was high in (But)2cAMP-treated cultures with short neuritic outgrowth but remained low in NGF- or NGF plus (But)2 cAMP-treated cultures, where neurite extension was, respectively, large and very large. These results suggest that excess proteolytic activity may be detrimental to neuritic outgrowth and that not only PAI release but also tPA-PAI complex formation is associated with production of large and stable neuritic outgrowth. This can be understood as an involvement of PAI in the protection against neurite-destabilizing proteolytic activity.