Potential role of new molecular plasma signatures on cardiovascular risk stratification in asymptomatic individuals

Abstract

The evaluation of cardiovascular (CV) risk is based on equations derived from epidemiological data in individuals beyond the limits of middle age such as the Framingham and SCORE risk assessments. Lifetime Risk calculator (QRisk®), estimates CV risk throughout a subjects’ lifetime, allowing those. A more aggressive and earlier intervention to be identified and offered protection from the consequences of CV and renal disease. The search for molecular profiles in young people that allow a correct stratification of CV risk would be of great interest to adopt preventive therapeutic measures in individuals at high CV risk. To improve the selection of subjects susceptible to intervention with aged between 30–50 years, we have employed a multiple proteomic strategy to search for new markers of early CV disease or reported CV events and to evaluate their relationship with Lifetime Risk. Blood samples from 71 patients were classified into 3 groups according to their CV risk (healthy, with CV risk factors and with a previously reported CV event subjects) and they were analyzed using a high through quantitative proteomics approach. This strategy allowed three different proteomic signatures to be defined, two of which were related to CV stratification and the third one involved markers of organ damage.

Unbiased plasma proteomics discovery of biomarkers for improved detection of subclinical atherosclerosis

“…Briefly, peptide concentration was measured using DirectDetect Infrared Spectrometer (Merck) and 50 µg of peptides from each sample were labeled. 23 , 24 After labeling, peptides from all the samples in each TMT experiment were mixed in the same tube, acidified with 25%TFA to a final concentration of 1% and desalted with Oasis cartridges (Waters). Aliquots of 1/10 (in volume) were saved for direct MS analysis (without peptide fractionation).…”

Why Does COVID-19 Affect Patients with Spinal Cord Injury Milder? A Case-Control Study: Results from Two Observational Cohorts

“…For the quantitative differential LC-MS/MS analysis using isobaric tags (TMT 10-plex), about 300 µg of total protein was digested using the filter-aided sample preparation (FASP) protocol described previously [ 15 ], with minor modifications. Samples were denatured by boiling for 5 min in the presence of 1% Sodium Dodecyl Sulfate (SDS) and 50 mM dithiothreitol (DTT), diluted in 7 M urea in 0.1 M Tris-HCl (pH 8.5: UA buffer) and loaded onto 10 kDa centrifugal filter devices (NanoSep 10k Omega, Pall Life Sciences, Port Washington, NY, USA).…”

Comprehensive Proteomic Profiling of Pressure Ulcers in Patients with Spinal Cord Injury Identifies a Specific Protein Pattern of Pathology

“…We used a previously described protocol, 14 with minor modifications. Quantitative differential LC-MS/MS analysis using TMT 10-plex isobaric labeling was developed with 100 lg of total protein, which was digested by the FASP protocol described previously, 15 with minor adjustments.…”

“…Labeled peptides were analyzed by LC-MS/MS as previously reported. 14 We used a C-18 reversed phase nano-column (75 lm I.D. • 50 cm, 2 lm particle size, Acclaim PepMap RSLC, 100 C18; Thermo Fisher Scientific, Waltham, MA) with a continuous ACN gradient consisting of 0-30% B in 360 min and 50-90% B in 3 min (A = 0.1% formic acid [FA]; B = 90% ACN, 0.1% FA).…”