Clinical Relevance and Methodological Comparison of Anti-MDA5 Antibody Detection: A Five-Year Retrospective and Exploratory Pilot Study

ObjectiveAutoantibodies to melanoma differentiation-associated gene 5 (MDA5) are specifically expressed in patients with dermatomyositis (DM) and are associated with a subset of DM patients with rapidly progressive interstitial lung disease (RP-ILD). Here, we examined the clinical utility of a newly developed enzyme-linked immunosorbent assay (ELISA) system for detecting these antibodies.MethodsHere we developed an improved ELISA for detecting anti-MDA5 antibodies. We then performed a multicenter clinical study involving 8 medical centers and enrolled 242 adult patients with polymyositis (PM)/DM, 190 with non-PM/DM connective tissue disease (CTD), 154 with idiopathic interstitial pneumonia (IIP), and 123 healthy controls. Anti-MDA5 antibodies in the patients’ serum samples were quantified using our newly developed ELISA, and the results were compared to those obtained using the gold-standard immunoprecipitation (IP) assay. In addition, correlations between the ELISA-quantified anti-MDA5 antibodies and clinical characteristics were evaluated.ResultsIn patients with PM/DM, the anti-MDA5 antibody measurements obtained from the ELISA and IP assay were highly concordant; the ELISA exhibited an analytical sensitivity of 98.2%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 99.5% (compared to the IP assay). Anti-MDA5 antibodies were detected in 22.7% of the DM patients, but not in any of the patients with PM, non-PM/DM CTD, or IIP. Clinically amyopathic DM, RP-ILD, arthritis, and fever were more prevalent in DM patients who were anti-MDA5 antibody-positive than in those who were antibody-negative (P ≤ 0.0002 for all comparisons). In addition, anti-MDA5 antibody-positive patients with RP-ILD exhibited higher antibody levels than those without RP-ILD (P = 0.006).ConclusionOur newly developed ELISA can detect anti-MDA5 antibodies as efficiently as the gold standard IP assay and has the potential to facilitate the routine clinical measurement of anti-MDA5 antibodies in patients who suspected to have DM.

Vaccine-induced thrombotic thrombocytopenia following Ad26.COV2.S vaccine in a man presenting as acute venous thromboembolism.

This study aimed to detect the expression of progranulin (PGRN) and elucidate associations with clinical features in dermatomyositis (DM) patients with anti-melanoma differentiation-associated gene 5 (anti-MDA5) antibody. We enrolled 40 DM patients with anti-MDA5 antibody, 20 patients with antisynthetase syndrome (ASS; disease control), and 20 healthy individuals (healthy control, HC). The clinical features of patients with anti-MDA5 antibody and anti-histidyl-tRNA antibody were collected. The level of PGRN in the serum was tested by ELISA. The PGRN levels in DM patients with anti-MDA5 antibody (166.74 ± 97.95ng/ml) were significantly higher than those in patients with ASS (82.66 ± 40.50ng/ml; p < 0.001) and in HC (42.34 ± 18.69ng/ml; p < 0.001). Patients with rapid progressive interstitial lung disease (RP-ILD) in DM with anti-MDA5 antibody (213.57 ± 114.05ng/ml) had higher levels of PGRN than those without RP-ILD (135.51 ± 72.41ng/ml; p = 0.012). ROC analysis showed an AUC value at 0.715 (95% CI, 0.541-0.888) for diagnosis of RP-ILD in DM patients with anti-MDA5 antibody. The expression of PGRN was positively correlated with the levels of ALT, AST, CK, LDH and ferritin (all p < 0.05). Our results indicated PGRN had great potential as a valuable serum marker of RP-ILD in DM with anti-MDA5 antibody. Key Points The level of PGRN was elevated in DM patients with anti-MDA5 antibody, especially for those with RP-ILD.

ABSTRACTObjectivesThe objectives of this exploratory study were to describe the safety, comfort, and effects of applying a weighted blanket to restrict anterior chest movement on oxygenation, and regional ventilation in clinically stable, nonventilated infants, and children hospitalized with lower respiratory tract infections (LRTIs). Our hypothesis is that the application of a weighted blanket would not adversely affect comfort levels or oxygenation in nonventilated infants and children with LRTIs.MethodA prospective, exploratory interventional pilot study of nonventilated children and infants aged 2 months to 9 years of age admitted to hospital for the management of a LRTI. Comfort and pain scores, vital signs (heart rate [HR], respiratory rate [RR], oxygen saturation, and blood pressure), and electrical impedance tomography (EIT) measurements were captured at baseline, immediately and at 5 min, 10 min, and 30 min after application of the blanket.ResultsThirty children (median interquartile range [IQR] 8.31 months old [2.46–22.27]; 16 [53.30%] males), received the intervention, and were included in the analysis. Just over half of the children recruited were admitted with a clinical diagnosis of pneumonia (n = 17; 56.70%). No adverse events were recorded during the study intervention. Overall, oxygen saturation increased from median (IQR) 97% (91–100) to 100% (93–100) (p < 0.001); COMFORT‐B score improved from median (IQR) 13 (6–14) to 11 (6–16) (p = 0.02) from baseline to 30 min postintervention and there was no significant change in the faces, legs, activity, cry, and consolability (FLACC) pain scale.ConclusionThe application of a weighted blanket in nonventilated infants and children with mild LRTI was well tolerated, with no adverse effects and was associated with improvements in comfort levels and SpO2.

Exergames integrate motor, cognitive, and postural-control demands; however, how specific task-demand manipulations influence gaze allocation during exergame performance remains insufficiently characterized. This exploratory pilot study used a within-participant design to examine whether gaze allocation and task performance differed across baseline, concurrent cognitive-task, and unilateral-squat conditions during squat-based exergaming in healthy young adults. Eight healthy adults (mean age 20, SD 1 years; 7 men and 1 woman) used a squat-based exergame (Ring Fit Adventure; Nintendo) under 3 randomized conditions: baseline, concurrent cognitive task, and unilateral squats. In the concurrent cognitive task condition, participants performed serial subtraction during squatting. In the unilateral-squat condition, participants performed single-leg squats, which were intended to increase support-leg muscular demand as well as postural and motor-control requirements. Execution time, squat score, arithmetic performance, and eye-tracking metrics were recorded. Primary gaze outcomes were the proportion of fixation time and fixation counts allocated to predefined areas of interest (AOIs; command, avatar, and score) and to regions outside these areas across the entire trial. Differences from the baseline condition were examined using Dunnett tests or paired Wilcoxon signed-rank tests with Holm adjustment, and effect sizes were reported. Exergame execution time increased from 31.0 (SD 3.3) seconds in the baseline condition to 38.2 (SD 6.1) seconds in the concurrent cognitive task condition and to 34.3 (SD 6.0) seconds in the unilateral-squat condition. In contrast, the squat score decreased from 98.8 (SD 2.8) in the baseline condition to 69.6 (SD 18.3) in the unilateral-squat condition. For fixation counts, allocation to the command AOI decreased from 44.8% (SD 11.4%) in the baseline condition to 32.1% (SD 8.9%) in the concurrent cognitive task condition and 36.2% (SD 8.5%) in the unilateral-squat condition. Outside AOI fixation counts increased in the concurrent cognitive task condition (55.7%, SD 8.6%) relative to the baseline condition (36%, SD 16.9%). In this exploratory within-participant pilot study, adding a concurrent cognitive task and performing unilateral squats resulted in different patterns of performance change and fixation count redistribution during squat-based exergaming. These preliminary findings suggest that AOI-based gaze allocation metrics may help characterize task demand-related attentional shifts in this setting. Larger confirmatory studies with more diverse samples and individual-level validation are needed before these metrics can be considered for adaptive or clinical applications.

The purpose of this exploratory and descriptive pilot study was to explore the process of school re-entrance for adolescents receiving outpatient treatment for cancers, using grounded theory. All adolescents who had received a minimum of 3 weeks' hospital-based schooling, but had since returned to their own schools, and were booked for an outpatient appointment on a paediatric oncology unit during a specified 2-week period, took part in an audio-tape-recorded, semi-structured interview. The sample consisted of five such adolescents. Data were analysed using the constant comparison method of grounded theory. Certain recognizable trends were noted leading to the development of a core variable to which other categories and their properties related. Central categories identified were 'disruption', 'adaptation' and ,normality', with this latter concept considered to be the core variable. Findings highlighted problem areas which are discussed and issues requiring further detailed investigation. This led to an undertaking of theoretical sampling into hospital-based schooling, as well as suggesting theoretical sampling for future studies.

A 69-year-old woman was admitted to our hospital due to a two-week history of low-grade fever, dry cough and erythematous eruptions. Physical examination revealed facial erythema, Gottron’s papules, mechanic’s hands and fine crackles in both lower lung fields on admission. Muscle weakness, a heliotrope rash and the shawl sign were not observed. Chest X-ray and computed tomography showed rapidly progressive interstitial lung disease (RP-ILD) with focal areas of air-space consolidation and linear opacities bilaterally. Antimelanoma differentiation-associated gene five (anti-MDA5) antibody levels were elevated at 241 index. Although lacking clinical myositis, the skin lesions and interstitial lung disease suggested the development of dermatomyositis. She was diagnosed with RP-ILD in clinically amyopathic dermatomyositis, we started initial combination therapy with high-dose glucocorticoids, a calcineurin inhibitor and intravenous cyclophosphamide. However, she died on hospital day 15 despite aggressive immunosuppressive therapy. On autopsy, pathological findings of the lungs included diffuse alveolar damage in the exudative phase with hyaline membranes, oedema and interstitial acute inflammation. Moreover, we examined titres of anti-MDA5 antibody over time using a series of frozen serum samples before the onset of RP-ILD and found detectable low titres of anti-MDA5 antibody more than 2 years before disease onset by enzyme-linked immunosorbent assay and more than 4 years before disease onset by immunoprecipitation.

Sustainability is not only a moral compass for organizations of all kinds and shapes; increasingly it can be identified as social representation influencing social practices and behavior. Conceptually inspired by the idea of preferences, conventions, and moral convictions influencing individual behavior and an innovative concept of moral harmonization strategies, this paper introduces a theoretical framework for the conceptualization of moralization effects in sustainability communication in general and food choices in particular. The framework is linked to empirical data from an exploratory qualitative pilot study, in which we conducted guideline-based interviews with 25 international students to gather information on individual perceptions of food choices and eating behaviors. Interview data were analyzed using inductive category formation to explore what role sustainability plays on an individual level in terms of coming in as a value or norm and how much sustainability as a normative principle influences individual decision-making processes and behavior. Based on the results of the pilot study, we hypothesize that food is less “morally overloaded” than expected and sustainability is not a moral imperative related to specific eating behavior. In line with previous findings, our results confirmed that food choices and changes in meat consumption involve a multi-faceted and complex decision-making process, which among others may be heavily influenced by inherent social norms within a person's social network, including family, friends as well as important other peers. Thus, with this preliminary study, we critically challenge existing literature on the influence of sustainability as moral imperative guiding and influencing individual behavior, at least in the domain of food and eating behavior. To elaborate on our proposed framework, additional empirical research is needed from a cultural, sustainability, language, and communication perspective.

To assess the utility of anti-melanoma differentiation-associated gene 5 (anti-MDA5) antibody measurement for predicting a risk for developing rapidly progressive interstitial lung disease (RP-ILD) in patients with polymyositis/dermatomyositis (PM/DM). A single-center cohort of 64 consecutive Chinese patients with PM/DM was examined. Serum anti-MDA5 antibody was measured by enzyme-linked immunosorbent assay. For meta-analysis, we searched PubMed and the Institute for Scientific Information Web of Knowledge for original studies that measured anti-MDA5 antibodies in patients with PM/DM. We calculated pooled sensitivity, specificity, diagnostic odds ratio (DOR), and the summary receiver operating characteristic (sROC) curve. In Chinese patients, anti-MDA5 antibodies were detected in 26 patients with classic DM or clinically amyopathic DM (CADM). Compared with anti-MDA5-negative patients, anti-MDA5-positive patients showed a higher prevalence of RP-ILD (P = 0.001). In a total of 233 patients with anti-MDA5 antibody, derived from 16 studies, a higher frequency of CADM was found in Japanese than in non-Japanese patients (74.7% versus 39.2%; P = 1.2 × 10(-7) ). Meta-analysis revealed that the pooled sensitivity and specificity of anti-MDA5 antibody for RP-ILD was 77% (95% confidence interval [95% CI] 64-87%) and 86% (95% CI 79-90%), respectively. The pooled DOR was 20.41 (95% CI 9.02-46.20) with a favorable area under the sROC curve of 0.89 (95% CI 0.63-0.98). Detection of anti-MDA5 antibody is a valuable tool for identifying DM patients with a high risk for developing RP-ILD, but the distribution of classic DM and CADM in patients with this antibody varies among ethnic groups.

Background: Anti-mitochondrial antibodies (AMA) are found in &gt;90% of primary biliary cholangitis patients. Anti-rods/rings antibodies (anti-RR) are most commonly associated with interferon-α and ribavirin treatment in hepatitis C patients. Clinical laboratories routinely screen for AMA and anti-RR using indirect immunofluorescence on HEp-2 cells (HEp-2-IFA). Therefore, we sought to establish reference materials for use in AMA and anti-RR testing. Methods: AMA-positive and anti-RR-positive human plasma samples (AMA-REF and RR-REF), identified as potential reference materials based on preliminary data, were further validated by multiple laboratories using HEp-2-IFA, immunoprecipitation (IP), western blotting, IP-western, line immunoassay (LIA), addressable laser bead immunoassay (ALBIA) and enzyme-linked immunosorbent assay (ELISA). Results: AMA-REF showed a strong positive cytoplasmic reticular/AMA staining pattern by HEp-2-IFA to ≥1:1280 dilution and positive signal on rodent kidney/stomach/liver tissue. AMA-REF reacted with E2/E3, E3BP, E1α and E1β subunits of the pyruvate dehydrogenase complex by IP and western blotting and was positive for AMA antigens by LIA, ALBIA and ELISA. RR-REF showed a strong positive rods and rings staining pattern by HEp-2-IFA to ≥1:1280 dilution. RR-REF reacted with inosine monophosphate dehydrogenase by IP, IP-western and ALBIA. RR-REF also produced a nuclear homogenous staining pattern by HEp-2-IFA, immunoprecipitated proteins associated with anti-U1RNP antibody and reacted weakly with histones, nucleosomes, Sm and nRNP/Sm by LIA. Conclusions: AMA-REF and RR-REF are useful reference materials for academic or commercial clinical laboratories to calibrate and establish internal reference standards for immunodiagnostic assays. AMA-REF and RR-REF are now available for free distribution to qualified laboratories through Plasma Services Group.

Anti-melanoma differentiation-associated gene5 (MDA5) antibody has been recognized to be significantly associated with a subset of dermatomyositis patients with rapidly progressive interstitial lung disease (RP-ILD). To elucidate the clinical characteristics and poor prognostic factors in Japanese dermatomyositis patients with anti-MDA5 antibody. Clinical features of anti-MDA5 antibody-positive dermatomyositis patients and risk factors, potentially associated with a poor prognosis, were retrospectively analysed. A total of 37.3% (28/75) dermatomyositis patients were positive for anti-MDA5 antibody. The frequency of Gottron's papules, palmar violaceous macules, antihelix/helix violaceous macules, and clinically amyopathic dermatomyositis (CADM) was significantly higher in patients with anti-MDA5 antibody. Of anti-MDA5 antibody-positive dermatomyositis patients, 57.1% developed RP-ILD, and in those with RP-ILD, serum ferritin level was markedly high and partial pressure of arterial oxygen (PaO2) was significantly low at first visit. Among patients with anti-MDA5 antibody with RP-ILD, non-survivors were older and revealed lower PaO2 at first visit relative to survivors. Furthermore, patients who did not take triple therapy (prednisolone, calcineurin inhibitor and cyclophosphamide) as initial treatment resulted in poor outcome. Anti-MDA5 antibody may be associated with CADM, the progression to RP-ILD, high serum ferritin level, and characteristic skin manifestations. High serum ferritin level in patients with anti-MDA5 antibody may be associated with the development of RP-ILD and poor prognosis. Early treatment with triple therapy, including intravenous cyclophosphamide, may improve the prognosis of RP-ILD.

The purpose of this study was to explore the clinical and serological features of patients with pneumomediastinum (PNM) and dermatomyositis-associated interstitial lung disease (DM-ILD). A total of 93 patients (68 with classic DM and 25 with clinically amyopathic DM [CADM]) were recruited. Clinical and laboratory data were collected retrospectively. Anti-melanoma differentiation-associated gene 5 (MDA5) antibodies were detected using enzyme-linked immunosorbent assay (ELISA). Variables were compared between patients with and those without PNM. Multivariate analysis was performed using a multivariate logistic regression model. A total of 11 patients developed spontaneous PNM. During the follow-up period, 6 patients died of respiratory failure. No differences in sex, age at the onset of DM, serum ferritin levels, and C-reactive protein (CRP) levels were observed between DM patients with and without PNM. Compared with DM patients without PNM, those with PNM had significantly higher frequencies of rapidly progressive ILD (RP-ILD) (63.6 vs 24.4 %, P = 0.01), anti-MDA5 antibodies (90.9 vs 52.4 %, P = 0.02), CADM diagnoses (63.6 vs 22.0 %, P = 0.007) and cutaneous ulcers (36.4 vs 11 %, P = 0.04), but significantly lower creatine kinase (CK) levels (58.5 vs 284 U/l, P = 0.04). The multivariate analysis indicated that cutaneous ulcer was the only independent risk factor for the occurrence of PNM in DM (OR = 5.98, 95 % confidence interval [CI] 1.12-31.98, P = 0.037). PNM is a refractory complication and tends to occur in DM patients with RP-ILD, anti-MDA5 antibody, CADM diagnosis, and low CK level, and especially in patients with cutaneous ulcers.

With the widespread use of immune checkpoint inhibitors to treat various cancers, pulmonary toxicity has become a topic of increasing concern. Anti-melanoma differentiation-associated gene 5 (anti-MDA5) antibodies are strongly associated with rapidly progressive interstitial lung disease (RP-ILD) in patients with clinically amyopathic dermatomyositis. However, anti-MDA5 antibody expression has not been reported in patients with immune-related adverse events. We present the case of a 74-year-old man with lung adenocarcinoma who developed RP-ILD after treatment with immune checkpoint inhibitors. Further investigation revealed multiple autoantibodies, including anti-MDA5 antibodies. He initially responded to systemic glucocorticoids, immunosuppressants, and tocilizumab but eventually died from worsening pneumomediastinum. This case is the first one to suggest that checkpoint inhibitor pneumonitis can present as RP-ILD with positive anti-MDA5 antibodies, which may be predictive of a poor prognosis.

SAT0185 Characteristics of Cytokine Profiles and Prognositc Factors in Interstitial Lung Disease with Polymyositis/Dermatomyositis

Clinically amyopathic dermatomyositis with interstitial lung disease double-positive for anti-MDA5 and anti-PL12 antibodies