OBJECTIVE:The aim of this study was to examine the association between peri-operative myocardial injury (MIn) and the occurrence of adverse cardiovascular events and or death one year after carotid revascularisation. METHODS:In this prospective, multicentre cohort study, 527 consecutive patients undergoing elective carotid endarterectomy (CEA) or carotid artery stenting (CAS) (June - October 2023) were enrolled across five tertiary centres. High sensitivity cardiac troponin was measured pre- and post-operatively, and patients were followed for one year. The primary endpoint was myocardial infarction (MI). Secondary endpoints were stroke, cardiac related death, and all cause death one year following CEA or CAS. Survival analysis was used to assess the impact of MIn on time dependent outcomes of interest. RESULTS:One year follow up was completed in 505 patients (95.8%), predominantly men (n = 349, 69.1%), asymptomatic (n = 339/504, 67.3%), with a mean age of 71.6 ± 8.6 years (range 41 - 90 years). During follow up, eight lethal outcomes were documented, of which one patient (0.2%) died due to cardiac related cause. One year post-operatively, the incidence of MI was 2.6% (n = 13) and the incidence of stroke was 3.8% (n = 19). Post-operative MIn and pre-operative diagnosis of malignancy independently predicted MI within one year after CEA or CAS (odds ratio [OR] 6.54, 95% confidence interval [CI] 1.91 - 22.22, p = .003; OR 6.13, 95% CI 1.67 - 22.73, p = .006, respectively). MI free survival was statistically significantly lower in patients with post-operative MIn one year after CEA or CAS (91.9% vs. 98.2%, log rank p = .003). CONCLUSION:Post-operative MIn independently predicted one year MI following CEA or CAS. Patients with post-operative MIn after carotid revascularisation represent a high risk subgroup in whom targeted strategies should be considered in order to reduce the risk of subsequent cardiac adverse events.

Background and aims: We have previously discovered that Proprotein Convertase Subtilisin/Kexin type 6 (PCSK6) plays a key role in vascular disease by smooth muscle cell (VSMC) migration, proliferation, and extracellular matrix (ECM) remodeling via MMP2/MMP14 axis. Transcriptomic data from the Stockholm abdominal aortic aneurysm (AAA) biobank demonstrated elevated PCSK6, especially in the medial layer, while histology showed its presence in T cells. This study aimed to investigate the role and regulatory mechanisms of Pcsk6 in AAA further. Methods and Results: Bulk RNA-seq AAA data from the Munich aortic biobank confirmed the upregulation of PCSK6 in AAA tissues, in particular in patients with ruptures. Human scRNA-seq data from AAAs showed strong PCSK6 expression in fibroblasts, T cells and macrophages, while mouse scRNA-seq data detected it in multiple cell types, most notably ECs, VSMCs, fibroblasts, T cells and macrophages. Functional in vivo studies were therefore performed using 1) peri-adventitial porcine pancreatic elastase (PPE) and 2) Ang II infusion-induced AAA models in male global Pcsk6 -/- mice, VSMC-ablated Tagln cre/cre Pcsk6 fl/fl mice, as well as these mice on ApoE -/- background and appropriate controls. After 4 weeks of AAA diameter follow-up with ultrasound, abdominal and thoracic aorta tissues were harvested for RNA-seq profiling and histology. Although there was no difference in baseline aortic function between Pcsk6 ablated vs. control mice by ex vivo myograph studies, ultrasound imaging showed that VSMC-ablated mice manifested larger AAAs with increased rupture rates. Mechanistically, AAA tissues from VSMC-ablated Pcsk6 mice revealed a reduction in collagen abundance, but remarkably increased T-cell and macrophage infiltration, as well as higher expression of Mmp2, Mmp14, and Il6. Moreover, RNA-seq data showed significantly elevated levels of platelet glycoprotein V (GP5) in VSMC Pcsk6 deficient aneurysms, indicating platelet and coagulant dysfunction. Conclusion: Aortic Pcsk6 plays an essential role in VSMC activation, ECM production, inflammation inhibition, as well as platelet and coagulant function, which are critical processes to prevent AAA progression and rupture.

The present study explored the interaction between genotype (G), environment (E), and their interaction (G x E) in determining grain iron content (GFeC), grain zinc content (GZnC), and grain yield per plot (GYPP) in wheat. Thirty-two genotypes were evaluated across eight contrasting environments, revealing significant phenotypic variation for all traits. GGE biplot and AMMI analyses identified Genotypes 14 ('EU 60') and 15 ('EU 61') as stable performers for micronutrient content, while Genotype 27 ('MP 1358') exhibited superior yield stability. A total of 113 significant marker-trait associations (MTAs) were detected, including 42 for GFeC, 23 for GZnC, and 48 for GYPP. Notably, marker trait association (MTAs) Xgwm468.1 and Xgwm538.1 were consistently associated with both GFeC and GZnC, while Xwmc382.4 was common to GFeC and GYPP, suggesting pleiotropy or tight linkage. Marker Xbarc74-5B was validated for GZnC, confirming its reliability for marker-assisted selection. These findings provide valuable genomic resources and phenotypic insights for future breeding programs aimed at developing high-yielding, climate-resilient, and nutrient-rich wheat cultivars.

OBJECTIVE:Despite elective repair of a large portion of stable abdominal aortic aneurysms (AAAs), the diameter criterion cannot prevent all small AAA ruptures. Since rupture depends on many factors, this study explored whether machine learning (ML) models (logistic regression [LogR], linear and non-linear support vector machine [SVM-Lin and SVM-Nlin], and Gaussian Naïve Bayes [GNB]) might improve the diameter based risk assessment by comparing already ruptured (diameter 52.8 - 174.5 mm) with asymptomatic (diameter 40.4 - 95.5 mm) aortas. METHODS:A retrospective case-control observational study included ruptured AAAs from two centres (2010 - 2012) with computed tomography angiography images for finite element analysis. Clinical patient data and geometric and biomechanical AAA properties were fed into ML models, whose output was compared with the results from intact cases. Classifications were explored for all cases and those having diameters below 70 mm. All data trained and validated the ML models, with a five-fold cross-validation. SHapley Additive exPlanations (SHAP) analysis ranked the factors for rupture identification. RESULTS:One hundred and seven ruptured (20.6% female, mean age 77 years, mean diameter 86.3 mm) and 200 non-ruptured aneurysmal infrarenal aortas (21.5% female, mean age 74 years, mean diameter 57 mm) were investigated through cross-validation methods. Given the entire dataset, the diameter threshold of 55 mm in males and 50 mm in females provided a 58.0% accurate rupture classification. It was 99.1% sensitive (AAA rupture identified correctly) and 36.0% specific (intact AAAs identified correctly). ML models improved accuracy (LogR 90.2%, SVM-Lin 89.5%, SVM-Nlin 88.7%, and GNB 86.4%); accuracy decreased when trained on the ≤ 70 mm group (55/50 mm diameter threshold 44.2%, LogR 82.5%, SVM-Lin 83.6%, SVM-Nlin 65.9%, and GNB 84.7%). SHAP ranked biomechanical parameters other than the diameter as the most relevant. CONCLUSION:A multiparameter estimate enhanced the purely diameter-based approach. The proposed predictability method should be further tested in longitudinal studies.

BACKGROUND:MicroRNAs are post transcriptional modulators of gene expression. We explored the diagnostic and prognostic value of circulating microRNAs in abdominal aortic aneurysm (AAA) disease, for which currently no established circulating biomarker is available. METHODS:We profiled the expression of 754 human microRNAs in plasma from 187 patients with AAA and 190 matched non-diseased controls. To validate, we used two additional AAA patient cohorts, looking at circulating and aortic tissue-derived microRNA expression, and their correlation to AAA disease phenotype, as well as two murine AAA models. RESULTS:We show that among 12 differentially expressed microRNAs, miR-15a and -659 are the most significantly up-regulated in AAA, whereas miR-1183 and -192 are the most significantly down-regulated. miR-15a is upregulated AAA patient tissues, and in plasma from two murine AAA models. In patients from three different cohorts, miR-15a expression levels in plasma, serum and aortic tunica media are significantly correlated with AAA diameter. Through modulation of miR-15a in human aortic smooth muscle cells, we identify several potential target genes of miR-15a known to be down-regulated in human AAA, suggesting its potential involvement in AAA pathology. Inhibition of miR-15a in vivo demonstrates a significant inhibition of murine aortic diameter growth at day 7. CONCLUSIONS:Our findings suggest that miR-15a is a potential biomarker of AAA. Through in vivo studies and based on its target profile, we show that miR-15a is involved in AAA pathogenesis and could help treatment, but also assist in risk-stratification of AAA patients and identify candidates for early AAA repair.

Aneurysm rupture is a life-threatening event, yet its underlying mechanisms remain largely unclear. This study investigated the fracture properties of the thoracic aneurysmatic aorta (TAA) using the symmetry-constraint Compact Tension (symconCT) test and compared results to native and enzymatic-treated porcine aortas’ tests. With age, the aortic stiffness increased, and tissues ruptured at lower fracture energy $$D$$ . Patients with bicuspid aortic valves were more sensitive to age, had stronger aortas and required more $$D$$ than tricuspid valves individuals (peak load: axial loading 4.42 $$\pm$$ 1.56 N vs 2.51 $$\pm$$ 1.60 N; circumferential loading 5.76 $$\pm$$ 2.43 N vs 4.82 $$\pm$$ 1.49 N. Fracture energy: axial loading 1.92 $$\pm$$ 0.60 kJ m-2 vs 0.74 $$\pm$$ 0.50 kJ m-2; circumferential loading 2.12 $$\pm$$ 2.39 kJ m-2 vs 1.47 $$\pm$$ 0.91 kJ m-2). Collagen content partly explained the variability in $$D$$ , especially in bicuspid cases. Besides the primary crack, TAAs and enzymatic-treated porcine aortas displayed diffuse and shear-dominated dissection and tearing. As human tissue tests resembled enzymatic-treated porcine aortas, microstructural degeneration, including elastin loss and collagen degeneration, seems to be the main cause of TAA wall weakening. Additionally, a tortuous crack developing during the symconCT test reflected intact fracture toughening mechanisms and might characterize a healthier aorta.

PURPOSE:The goal of the study described in this protocol is to build a multimodal artificial intelligence (AI) model to predict abdominal aortic aneurysm (AAA) shrinkage 1 year after endovascular aneurysm repair (EVAR). METHODS:In this retrospective observational multicenter study, approximately 1000 patients will be enrolled from hospital records of 5 experienced vascular centers. Patients will be included if they underwent elective EVAR for infrarenal AAA with initial assisted technical success and had imaging available of the same modality preoperatively and at 1-year follow-up (CTA-CTA or US-US). Data collection will include baseline and vascular characteristics, medication use, procedural data, preoperative and postoperative imaging data, follow-up data, and complications. PROPOSED ANALYSES:The cohort will be stratified into 3 groups of AAA remodeling based on the maximum AAA diameter difference between the preoperative and 1-year postoperative moment. Patients with a diameter reduction of ≥5 mm will be assigned to the AAA shrinkage group, cases with an increase of ≥5 mm will be assigned to the AAA growth group, and patients with a diameter increase or reduction of <5 mm will be assigned to the stable AAA group. Furthermore, an additional fourth group will include all patients who underwent an AAA-related reintervention within the first year after EVAR, because both the complication and the reintervention might have influenced the state of AAA remodeling at 1 year. The preoperative and postoperative CTA scans will be used for anatomical AAA analysis and biomechanical assessment through semi-automatic segmentation and finite element analysis. All collected clinical, biomechanical, and imaging data will be used to create an AI prediction model for AAA shrinkage. Explainable AI techniques will be used to identify the most descriptive input features in the model. Predicting factors resulting from the AI model will be compared with conventional univariate and multivariate logistic regression analyses to find the best model for the prediction of AAA shrinkage. The study is registered at www.clinicaltrials.gov under the registration number NCT06250998. CLINICAL IMPACT:This study aims to develop a robust and high-performance AI model for predicting AAA shrinkage one-year after EVAR, with great potential for optimizing both EVAR treatment and follow-up. The model can identify cases with an initially lower chance of early AAA shrinkage, in whom EVAR-treatment could be tailored by including additional preoperative coil embolization, active sac management and/or postoperative tranexamic acid therapy, which have shown to promote AAA shrinkage rate but are too complex and costly to perform in all patients. The model could aid in stratification of post-EVAR surveillance based on the patient's individual risk and possibly decrease follow-up for the 40-50% of patients who will experience AAA sac shrinkage. Overall, the AI prediction model is expected to improve patient survival and decrease the number of reinterventions after EVAR and associated healthcare costs.

Objective:Carotid endarterectomy for symptomatic carotid stenosis is recommended for patients with >70% stenosis, but not in those with <50%. Because non-significant, low-degree stenoses may still cause strokes, refined risk stratification is necessary, which could be improved by assessing biological features of plaque instability. To challenge risk-stratification based on luminal narrowing, we compared biological features of carotid plaques from symptomatic patients with low-degree (<50%) vs high-degree (>70%) stenosis and explored potential mechanisms behind plaque instability in low-degree stenoses. Methods:Endarterectomy specimens were taken from symptomatic patients with high-degree (n = 204) and low-degree (n = 34) stenosis, all part of the Biobank of Karolinska Endarterectomies. Patient demographics, image-derived plaque morphology, and gene expression analyses of extracted lesions were used for comparisons. Plaque biology was assessed by transcriptomics using dimensionality reduction, differential gene expression, and gene-set enrichment analyses. Immunohistochemistry was used to study proteins corresponding to upregulated genes. Results:The demographics of the two groups were statistically similar. Calcification, lipid-rich necrotic core, intraplaque hemorrhage, plaque burden, and fibrous cap thickness were similar in both groups, whereas the sum of lipid-rich necrotic core and intraplaque hemorrhage was higher (P = .033) in the high-degree stenosis group. Dimensionality reduction analysis indicated poor clustering separation of plaque gene expression in low-compared with high-degree stenosis lesions, whereas differential gene expression showed upregulation of hypoxia-inducible factor 3A (log2 fold change, 0.7212; P = .0003), and gene-set enrichment analyses identified pathways related to tissue hypoxia and angiogenesis in low-degree stenoses. Hypoxia-inducible factor 3-alpha protein was associated with smooth muscle cells in neo-vascularized plaque regions. Conclusions:Plaques from symptomatic patients with non-significant low-degree carotid stenoses showed morphologic and biological features of atherosclerotic plaque instability that were comparable to plaques from patients with high-degree stenoses, emphasizing the need for improved stroke risk stratification for intervention in all patients with symptomatic carotid stenosis irrespective of luminal narrowing. An increased expression of hypoxia-inducible factor 3A in low-degree stenotic lesions suggested mechanisms of plaque instability associated with tissue hypoxia and plaque angiogenesis, but the exact role of hypoxia-inducible factor 3A in this process remains to be determined. Clinical relevance:Carotid plaques from symptomatic patients with <50% stenosis show morphologic and biological features of plaque instability, comparable to high-degree stenosis, which emphasizes the need for improved stroke risk stratification beyond stenosis severity.

BACKGROUND:Abdominal aortic aneurysms (AAAs) are characterized by ECM (extracellular matrix) degradation and chronic vascular inflammation, with macrophages playing a key role. The mechanisms regulating macrophage activation in AAA remain incompletely understood. Vascular macrophages express Olfr2 (olfactory receptor 2), a GPCR (G-protein-coupled receptor) implicated in inflammation, but its role in AAA development is unknown. METHODS:We investigated the role of Olfr2 in AAA using PPE (porcine pancreatic elastase) infusion in Olfr2-deficient (Olfr2-/-), Ang II (angiotensin II) infusion in Apoe-/- Olfr2-/-mice, bone marrow transplantation, and pharmacological modulation experiments. Echocardiography and histology were complemented by spectral flow cytometry, transcriptional profiling, and functional in vivo and ex vivo assays. RESULTS:Microarray analysis revealed increased expression of the human Olfr2 orthologue OR6A2 (olfactory receptor family 6 subfamily A member 2) in AAA tissue. Flow cytometry showed OR6A2 upregulation in monocytes from patients with large versus small AAAs. In both human and murine tissues, up to 30% of vascular macrophages expressed OR6A2/Olfr2, which peaked in MHCIIhigh CCR2low monocytes/macrophages on day 7 of experimental AAA. Both whole-body and hematopoietic Olfr2 deficiency protected mice from AAA formation, with reduced ECM degradation, decreased macrophage infiltration, and preserved smooth muscle cell content. Treatment with the Olfr2 agonist octanal exacerbated, while the antagonist citral reduced AAA and inflammation. In Olfr2-/- mice, inflammatory gene expression and aortic leukocyte accumulation were diminished. Despite a similar total leukocyte count, Ly6Chigh monocytes displayed reduced CX3CR1 (CX3C motif chemokine receptor 1) expression and impaired migration toward CX3CL1 in vitro. Competitive transfer confirmed reduced migratory capacity of Olfr2-/- monocytes, while pharmacological CX3CR1 inhibition mitigated the proinflammatory effects of octanal in AAA. CONCLUSIONS:Olfr2 regulates monocyte recruitment and macrophage-driven inflammation during AAA. Its genetic deletion or pharmacological inhibition protects against AAA, whereas receptor activation worsens the disease. Olfr2 represents a critical modulator of vascular inflammation and a potential therapeutic target in AAA.

Currently there is no effective pharmacotherapy to prevent the growth and rupture of abdominal aortic aneurysms. Using a mouse model that combines cigarette smoke exposure and hypercholesterolemia, we demonstrated that cigarette smoke exacerbated atherosclerosis, leading to elastin fragmentation, aneurysm formation, rupture and death. Arterial injury was driven by macrophages that accumulated within atherosclerotic plaques and exhibited tissue-degrading proteolytic activity in vivo (a process dependent on the endothelial cell-derived macrophage growth factor CSF-1). Single-nucleus RNA sequencing revealed that cigarette smoke-induced endothelial cell dysfunction promoted monocyte recruitment and inflammatory signaling and amplified vascular injury. Furthermore, single-cell transcriptomic analysis identified conserved macrophage responses across mouse and human abdominal aortic aneurysm, including TREM2+ macrophages, which were key mediators of arterial damage. These findings established atherosclerotic plaque macrophages as critical drivers of aneurysm pathology and provide key insights into the mechanisms underlying aneurysm progression and rupture.

OBJECTIVE:To assess the prevalence and risk factors for the development of myocardial injury (MIn) in patients undergoing carotid revascularisation and to assess whether elevated troponin levels can predict the occurrence of adverse cardiac events and short term death. METHODS:This prospective, multicentre cohort study included 527 patients subjected to carotid revascularisation from June to October 2023. High sensitive cardiac troponin I and or T were assessed pre-operatively, and eight and 24 hours post-operatively. Myocardial injury was defined as at least one value of cardiac troponin above the 99th percentile upper reference limit. RESULTS:Four hundred and forty patients (83.5%) underwent carotid endarterectomy and 87 (16.5%) carotid artery stenting (85 under local infiltrative and two under general anaesthesia). Of the 527 patients, 87 (16.5%) were operated under general, 355 (67.4%) under regional anaesthesia, and the remaining 85 (16.1%) received local anaesthetic infiltration. Pre-operatively, MIn was noted in 7% of patients, and the overall prevalence of post-operative MIn was 12.3%. Age ≥ 81 years, pre-operative haemoglobin 10 - 13.9 g/dL and glomerular filtration rate 15 - 30 mL/min/1.73 m2 were independent predictors of post-operative MIn (OR 2.84, 95% CI 1.17 - 6.91, p = .021; OR 1.95, 95% CI 1.01 - 3.76, p = .046; OR 11.46, 95% CI 2.37 - 55.33, p = .002, respectively). Myocardial infarction (MI) developed in seven patients (1.3%), more frequently in those who had MIn eight and 24 hours following surgery (71.4% vs. 10.4%, p = .001; 71.4% vs. 10.2%, p = .001, respectively). CONCLUSION:Myocardial injury is common in patients undergoing carotid revascularisation; to reduce the rate of MIn, special attention should be paid to those patients with risk factors identified in the present study. Long term (one and two year) follow up of the TROPICAR patients will provide additional insights into the association between peri-operative MIn, MI, and death.

Background: Abdominal aortic aneurysms (AAA) are characterized by an intricate interplay of extracellular matrix degradation and inflammation. Macrophages are centrally involved in these processes. The mechanisms underlying macrophage activation in AAA remain incompletely understood. Vascular macrophages have been shown to express olfactory receptor 2 (Olfr2), a G-protein coupled receptor involved in mediating the sense of smelling and regulating inflammatory activity in macrophages. Whether Olfr2 plays a role in modulating macrophage responses in AAA formation remains unknown. Methods & Results: In silico micro-array analysis showed increased expression of the human Olfr2 orthologue OR6A2 in AAA tissue compared to healthy aorta. Flow cytometric analysis revealed increased expression of OR6A2 on classical, non-classical, and intermediate monocytes of patients with a large AAA (> 5cm) in comparison to patients with smaller AAA (< 5cm). Up to 30% of vascular macrophages expressed OR6A2 in human AAA and Olfr2 in mouse AAA tissue. Olfr2 expression peaked on major histocompatibility complex II-high (MHCIIhigh) and C-C chemokine receptor type 2-low (CCR2low) aortic monocytes and macrophages on day 7 following experimental AAA initiation and decreased to baseline expression on day 28. Olfr2 gene (Olfr2-/-) deficiency protected mice from AAA formation, which was accompanied by lowered ECM degradation, reduced macrophage infiltration and increased smooth muscle cell content. Conversely, treatment with the Olfr2 agonist octanal exacerbated AAA formation and inflammation, while the antagonist citral reduced AAA formation in comparison to vehicle treated mice. Bulk transcriptome analysis of aortic tissue revealed reduced inflammatory gene expression in Olfr2-/- mice at day 7 following AAA initiation. Spectral flow cytometry resolved 20 aortic immune cell populations, which were largely reduced in quantity by Olfr2-deficiency at day 7 and day 28 post experimental AAA formation, while circulating leukocyte counts and monocyte subset distribution were not altered between Olfr2+/+ and Olfr2-/- mice. Circulating Ly6Chigh-monocytes exhibited reduced expression of the CX3C motif chemokine receptor 1 (CX3CR1) and CCR2 during AAA formation. Transcriptional analysis of monocytes confirmed downregulation of pathways associated with cell adhesion, motility and migration. In vitro, Olfr2-/- monocytes showed impaired migration towards the CX3CR1 ligand CX3CL1. Competitive transfer of Olfr2+/+ and Olfr2-/- monocytes confirmed reduced migratory capacity of Olfr2-/- monocytes into the developing AAA. Conclusion: We demonstrate a critical relevance for Olfr2 in the modulation of the inflammatory response underlying AAA, which is mediated by enhanced monocyte recruitment. ### Competing Interest Statement The authors have declared no competing interest.

The human blood proteome provides a holistic readout of health states through the assessment of thousands of circulating proteins. In this study, we present a pan-disease resource to enable the study of diverse disease phenotypes within a harmonized proteomics dataset. By profiling protein concentrations across 59 diseases and healthy cohorts, we identified proteins associated with age, sex, and body mass index, as well as disease-specific signatures. This study highlights shared and distinct protein patterns across conditions, demonstrating the power of a unified proteomics approach to uncover biological insights. The dataset, covering 8262 individuals and up to 5416 proteins, serves as an online resource for exploring disease-specific protein profiles and advancing precision medicine research.