Rapid and accurate diagnosis of emerging inflammatory illnesses is challenging due to overlapping clinical features with existing conditions. We demonstrate an approach that integrates proteomic analysis with machine learning to identify diagnostic protein signatures, using the example of SARS-CoV-2-induced multisystem inflammatory syndrome in children (MIS-C). We used plasma samples collected from subjects diagnosed with MIS-C and compared them first to controls with asymptomatic/mild SARS-CoV-2 infection and then to controls with pneumonia or Kawasaki disease. We used mass spectrometry to identify proteins and support vector machine (SVM) algorithm-based classification schemes to identify protein signatures. Diagnostic accuracy was assessed by calculating sensitivity, specificity, and area under the ROC curve (AUC), and corrected for overfitting by cross-validation. Proteomic analysis of a training dataset containing MIS-C (N = 17), and asymptomatic/mild SARS-CoV-2 infected control samples (N = 20) identified 643 proteins, of which 101 were differentially expressed. Plasma proteins associated with inflammation increased, and those associated with metabolism and coagulation decreased in MIS-C relative to controls. The SVM machine learning algorithm identified a three-protein model (ORM1, AZGP1, SERPINA3) that achieved 90.0% specificity, 88.2% sensitivity, and 93.5% AUC, distinguishing MIS-C from controls in the training set. Performance was retained in the validation dataset utilizing MIS-C (N = 19) and asymptomatic/mild SARS-CoV-2 infected control samples (N = 10) (90.0% specificity, 84.2% sensitivity, 87.4% AUC). We next replicated our approach to compare MIS-C with similarly presenting syndromes, such as pneumonia (N = 17) and Kawasaki disease (N = 13), and found a distinct three-protein signature (VWF, FCGBP, and SERPINA3) that accurately distinguished MIS-C from the other conditions (97.5% specificity, 89.5% sensitivity, 95.6% AUC). A software tool was also developed that may be used to evaluate other protein signatures using our data. These results demonstrate that the use of mass spectrometry to identify candidate plasma proteins followed by machine learning, specifically SVM, is an efficient strategy for identifying and evaluating biomarker signatures for disease classification.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-20684-5.

This Viewpoint discusses different approaches to evaluating and managing melanoma in situ to help mitigate the potential harms of overdiagnosis.

We previously demonstrated that the NO-stimulated soluble guanylyl cyclase (GC1), which produces cGMP, also has the ability to transfer S-nitrosothiols (SNO) to other proteins in a reaction involving oxidized Thioredoxin 1 (oTrx1). This transnitrosation cascade was established in vitro and involved Cys 610 (C610) of GC1 as the major SNO-donor. To assay the relevance of GC1 transnitrosation under physiological conditions and in oxidative pathologies, we studied a knock-in mouse in which C610 was replaced with a serine (KI αC610S) under basal or angiotensin II (Ang II)-treated conditions. Despite similar GC1 expression and NO responsiveness, the Ang II-treated KI mice displayed exacerbated oxidative pathologies including higher mean arterial pressure and more severe cardiac dysfunctions compared to the Ang II-treated WT. These phenotypes were associated with a drastic decrease in global S-nitrosation and in levels of SNO-Trx1 in the KI mice. To investigate the mechanism underlying the dysregulation of blood pressure, pressure myography and in vivo intravital microscopy were conducted to analyze the vascular tone of resistance vessels. Both approaches indicated that, even in the absence of oxidative stress, the single mutation C610S led to a significant disruption of the endothelium-dependent, acetylcholine-induced vasorelaxation while NO-dependent smooth muscle relaxation remained unchanged. Mechanistically, the vasorelaxation defect was associated with decreased endothelial calcium influx and membrane hyperpolarization, independent of NO bioavailability. These findings indicate that the C610S mutation uncouples two NO signaling vasodilatory pathways (endothelial SNO and smooth muscle NO-cGMP) and suggest that GC1 transnitrosation activity is essential for endothelium-derived hyperpolarization.

ABSTRACTBackgroundCancer advocacy groups engage patients, families and caregivers in navigating the cancer landscape, with a focus on early detection/screening and providing psychosocial and financial support during and after treatment. These groups are influential among their communities, funders and policymakers.ObjectiveWe wished to understand perceptions of breast cancer advocacy group leaders on primary care's role in breast cancer survivorship care, given limited primary care engagement despite endorsement by the National Cancer Institute (NCI).MethodsAs part of a larger NCI‐funded study, we used purposive sampling to select leaders from a diversity of patient advocacy groups for in‐depth interviews (n = 9). After obtaining consent, interviews were conducted and recorded on Zoom, professionally transcribed, and analysed using an established immersion–crystallisation process to identify themes and patterns.ResultsWe interviewed leaders (n = 9) from two local, three regional and four national advocacy groups, five of whom had personal experiences with breast cancer. These advocates emphasised that transitions away from the safety of oncology to primary care are difficult for patients. They felt patients with a history of breast cancer have unique and complex needs that are different from the standards of care found within primary care settings, and primary care clinicians are not adequately prepared to address these. In reflecting on the ideal role of primary care, they highlighted listening to patients, identifying issues and referring patients to appropriate specialists, but ultimately stressed that patients needed to advocate for themselves in the current healthcare environment.ConclusionsAdvocacy groups typically start as grass root efforts motivated by perceptions of inadequate support and care for cancer patients. As such, there is potential for advocacy groups to shape the conversation to improve collaboration between oncology and primary care by articulating and advocating for better primary care involvement in survivorship care.Patient or Public ContributionThe project's steering committee included cancer survivors and cancer advocacy group leaders who provided feedback on the project design and made recommendations for people to interview. Steering committee retreats were later held after the completion of interviews to reflect on emerging findings and plan dissemination strategies. The study team included a cancer survivor and several members whose immediate family members had a history of cancer. They were actively engaged in the design, analysis and manuscript writing.

ABSTRACTObjectiveTo investigate the impact of clinical surveillance, primary radiotherapy, and primary surgery on overall survival (OS) in laryngeal carcinoma in situ (Cis).MethodsThe 2006–2020 National Cancer Database was queried for adults with a biopsy‐proven diagnosis of laryngeal Cis. Multivariable binary logistic and Cox proportional hazards regression models were implemented.ResultsOf 3567 unique patients satisfying inclusion criteria, 514 (14.4%) underwent clinical surveillance, 1074 (30.1%) underwent primary radiotherapy, and 1979 (55.5%) underwent primary surgery. Receiving treatment at an academic/research facility was associated with higher odds of undergoing primary surgery compared to primary radiotherapy. Among 646 patients undergoing primary surgery with known pT classification and margins, 570 (76.6%) had pTis and NSM and 174 (23.4%) had pT1 and/or PSM. 5‐year OS of clinical surveillance, primary radiotherapy, and primary surgery was 73%, 81%, and 86%, respectively (p < 0.001). Patients undergoing primary surgery with invasive or residual disease (i.e., pT1 and/or PSM) had similar 5‐year OS as those without (84% vs. 88%, p = 0.057). Compared with primary radiotherapy, clinical surveillance (aHR 1.29, 95% CI 1.06–1.57, p = 0.003) was associated with worse OS, and primary surgery (aHR 0.80, 95% CI 0.69–0.92, p = 0.003) was associated with higher OS.ConclusionPrimary surgery is associated with higher OS than clinical surveillance and primary radiotherapy among patients with laryngeal Cis.Level of Evidence4.