The impact of post-ovulatory ageing on the development of diploid and triploid Atlantic salmon (Salmo salar L.)

This study is the first to investigate the effects of post-ovulatory ageing (POA) on both diploid and triploid Atlantic salmon (Salmo salar L.). Following ovulation, female broodstock were partially stripped periodically (at 0, 5, 10, 15 and 20 days post-ovulation, DPO), creating five groups of ova with differing durations held in the body cavity. Survival and bone architecture were negatively impacted by increasing POA, irrespective of ploidy, and an increase in individual growth variation was observed, particularly in triploids. Several ovarian fluid and egg quality parameters were screened to determine potential prospective biomarkers that may predict the future success of an egg batch. Ovarian fluid osmolality and egg lipid composition, vitamin E concentration and TBARS emerged as good candidates. Furthermore, significant changes in abundance of 19 proteins involved in cell organisation, metabolism and reproduction were observed at different stages of POA. Also, a suite of genes involved in energy metabolism, apoptosis and cell cycle regulation showed significantly reduced expression in eggs at 20 DPO compared to 0 DPO. The data show that increasing POA negatively impacts egg survival and future growth and increases deformity prevalence. Several potential biomarkers show promise and should be investigated further. Although egg quality appears to remain stable up to 10 DPO, the data suggests that fertilising eggs within 5 DPO would reduce the variation in growth, thus improving future performance potential and further benefitting stock management of both diploid and triploid Atlantic salmon

Reproducibility Evaluation of Urinary Peptide Detection Using CE-MS

In recent years, capillary electrophoresis coupled to mass spectrometry (CE-MS) has been increasingly applied in clinical research especially in the context of chronic and age-associated diseases, such as chronic kidney disease, heart failure and cancer. Biomarkers identified using this technique are already used for diagnosis, prognosis and monitoring of these complex diseases, as well as patient stratification in clinical trials. CE-MS allows for a comprehensive assessment of small molecular weight proteins and peptides (<20 kDa) through the combination of the high resolution and reproducibility of CE and the distinct sensitivity of MS, in a high-throughput system. In this study we assessed CE-MS analytical performance with regards to its inter- and intra-day reproducibility, variability and efficiency in peptide detection, along with a characterization of the urinary peptidome content. To this end, CE-MS performance was evaluated based on 72 measurements of a standard urine sample (60 for inter- and 12 for intra-day assessment) analyzed during the second quarter of 2021. Analysis was performed per run, per peptide, as well as at the level of biomarker panels. The obtained datasets showed high correlation between the different runs, low variation of the ten highest average individual log2 signal intensities (coefficient of variation, CV < 10%) and very low variation of biomarker panels applied (CV close to 1%). The findings of the study support the analytical performance of CE-MS, underlining its value for clinical application

Association of fatal and non-fatal adverse health outcomes with urinary peptides reflecting collagen I turnover

Background Imbalance of collagen I (COL1) turnover, featured by increased synthesis and decreased degradation of collagen fibers, is a hallmark of fibrosis in the heart and blood vessels that associates with poor cardiovascular outcomes. Such as imbalance of COL1 turnover could be reflected in urine and serve as fingerprint for future adverse outcomes in general population, and high risk subjects. Purpose We hypothesize that imbalance of proteomic signatures of urinary peptides (UPs) reflecting COL1 turnover relate to adverse health outcomes in participants from a general population. Methods We randomly recruited 776 participants (51.2% women; 50.5 years) from the Flemish Study on Environment, Genes and Health Outcomes cohort and measured UPs proteome by capillary electrophoresis coupled with mass spectrometry. Our analyses focused on 148 peptides of COL1 alpha-1 (COL1A1) chain that retained ≥70% signal in the whole sample. The primary endpoint included fatal and nonfatal cardiovascular endpoints. Secondary endpoints consisted of total mortality, fatal and nonfatal cardiac, coronary, and heart failure endpoints. Multivariate Cox proportional models, partial least squares analysis (PLS), log-likelihood test, and receiver operating characteristics (ROC) curve were applied. Results Over a median follow up of 12.4 years, 110 primary endpoints occurred, 61 participants died, 81, 41 and 24 experienced cardiac, coronary, and heart failure endpoints; respectively. In PLS analyses, upregulation of UPs signatures closer to C- and N-terminal locations of the COL1A1 chain whereas downregulation of mid-region UPs were associated with lower risk of adverse health outcomes. This pattern was inverted in subjects with cardiovascular disease, as upregulation of terminal and downregulation of mid region UPs increased risk. Adding UPs to a basic model including sex, age and usual cardiovascular risk factors significantly improved model performance between 2.54% to 4.93% (P≤0.001) for prediction of adverse health outcomes. In ROC plots, adding UPs to the basic model increased the area under the curve up to 4.00% (P\u3c0.012). Conclusions UPs reflecting COL1 turnover predicted adverse health outcomes. The inverted up- and down regulations of UPs in between participants with and without previous cardiovascular diseases might be explained by a shift in the UPs signatures of COL1 fragments linked to distinct fibrotic processes. Urinary proteomic might have clinical importance in documenting the extent of collagen accumulation that relates to adverse health outcomes. In patients at high cardiovascular risk, modification of collagen I fibers turnover might be a potential treatment target. Funding Acknowledgement Type of funding sources: Public grant(s) – EU funding. Main funding source(s): The European Union the European Research Council and the European Research Area Net for Cardiovascular Diseases

Increased Collagen Turnover Is a Feature of Fibromuscular Dysplasia and Associated With Hypertrophic Radial Remodeling: A Pilot, Urine Proteomic Study.

Fibromuscular dysplasia (FMD), a nonatherosclerotic, noninflammatory disease of medium-sized arteries, is an underdiagnosed disease. We investigated the urinary proteome and developed a classifier for discrimination of FMD from healthy controls and other diseases. We further hypothesized that urinary proteomics biomarkers may be associated with alterations in medium-sized, but not large artery geometry and mechanics. The study included 33 patients with mostly multifocal, renal FMD who underwent in depth arterial exploration using ultra-high frequency ultrasound. The cohort was separated in a training set of 23 patients with FMD from Belgium and an independent test set of 10 patients with FMD from Italy. For each set, controls matched 2:1 were selected from the Human Urinary Proteome Database. The specificity of the classifier was tested in 700 additional controls from general population studies, patients with chronic kidney disease (n=66) and coronary artery disease (n=31). Three hundred thirty-five urinary peptides, mostly related to collagen turnover, were identified in the training cohort and combined into a classifier. When applying in the test cohort, the area under the receiver operating characteristic curve was 1.00, 100% specificity at 100% sensitivity. The classifier maintained a high specificity in additional controls (98.3%), patients with chronic kidney (90.9%) and coronary artery (96.8%) diseases. Furthermore, in patients with FMD, the proteomic score was positively associated with radial wall thickness and wall cross-sectional area. In conclusion, a proteomic score has the potential to discriminate between patients with FMD and controls. If confirmed in a wider and more diverse cohort, these findings may pave the way for a noninvasive diagnostic test of FMD