Cardiovascular Disease Biomarkers Research Articles

Innovative biomarkers in cardiovascular disease: Advances in prevention and therapeutic

Innovative biomarkers in cardiovascular disease: Advances in prevention and therapeutic

Serum megalin levels in type-2 diabetes mellitus with and without cardiovascular disease

ABSTRACT Background: Low-density lipoprotein receptor-related protein-2 (LRP2), also called megalin, is a multi-ligand receptor of the LDL receptor gene family mediating reabsorption of ligands like Apo-A1. Type 2 diabetes mellitus (T2DM) may possibly disrupt megalin functions as it is found to be regulated by insulin. This might cause cardiovascular complications due to derangement in lipoprotein metabolism. The current study was carried out to assess the serum megalin levels among T2DM individuals with cardiovascular complications in the Indian population. Methods: This was a cross-sectional study involving 80 patients with T2DM. 40 T2DM patients with known cardiovascular disease were selected as cases and 40 of those without evidence of cardiovascular disease were selected as controls. Demographic details, DM duration and class of oral hypoglycemic agents (OHA) used were collected from medical records while details of lipid profile, fasting glucose, serum creatinine and urea were collected from laboratory information system. Serum megalin levels were estimated using left-over samples by ELISA. Results: The study groups showed no statistical significance in baseline laboratory parameters except for serum creatinine and low density lipoprotein cholesterol (LDL-c). Mean serum megalin levels were statistically insignificant between cases and controls (0.91 ± 0.78 ng/mL vs. 0.85 ± 0.69 ng/mL, P 0.74).The subgroup analysis of serum megalin levels based on OHA consumption among cases was also statistically insignificant (P 0.056). Pearson’s correlational analysis was statistically insignificant between serum megalin and lipid profile parameters among cases (P > 0.05). Conclusion: Serum megalin alone may not serve as a biomarker for cardiovascular disease.

Serum neurofilament light chain: a novel biomarker for cardiovascular diseases in individuals without hypertension

Serum neurofilament light chain (sNFL) is a biomarker for axonal injury. Previous studies have linked sNFL levels to cardiovascular risk factors such as diabetes and hypertension, but its association with cardiovascular diseases (CVD) remains unclear. This study aims to explore the association between sNFL and CVD and evaluates its predictive value. Utilizing NHANES 2013–2014 data, this study included 2,035 participants aged ≥ 20 years with measured sNFL quantified using a Siemens immunoassay. CVD was self-reported and included myocardial infarction, stroke, heart failure, coronary heart disease, or angina. Logistic regression models assessed the association between sNFL levels and CVD. The predictive value of sNFL for CVD was evaluated using area under the curve (AUC) and DeLong test. Participants with higher sNFL levels were typically older, male, non-Hispanic white, smokers, and had lower socioeconomic status, higher CVD, hypertension, and diabetes prevalence. Higher sNFL levels were significantly associated with increased odds of CVD (adjusted OR = 1.41, 95% CI: 1.05–1.88). The association was significant in non-hypertensive individuals (OR = 2.72, 95% CI: 1.61–4.62) but not in hypertensive individuals (OR = 1.13, 95% CI: 0.81–1.56). sNFL addition to traditional risk models improved predictive accuracy, especially in non-hypertensive individuals (AUC from 0.827 to 0.856). sNFL levels are significantly associated with CVD in the general population, with a strong predictive value in non-hypertensive individuals. Future longitudinal studies should validate sNFL’s efficacy in various populations and explore the underlying mechanisms of its relationship with hypertension and CVD.

Circulating nucleases degrade high-molecular-weight cell-free DNA in coronary vessels during acute myocardial infarction

Abstract Introduction Extracellular cell-free DNA (cfDNA) originates from various biological processes, including cell turnover, apoptosis, necrosis, NETosis, and other forms of cell death. In healthy individuals, highly fragmented cfDNA is elevated in the bloodstream due to infection, cancer, inflammation and thrombosis. In thrombotic vascular diseases, neutrophils release elongated, high-molecular-weight DNA with bioactive and prothrombotic properties in a process termed NETosis. Circulating nucleases are thought to counteract pathogenic properties of NETs. However, attempts to connect total cfDNA concentration to nuclease activity have failed. Purpose We aimed to elucidate the in vivo connection between extracellular DNA and circulating nucleases in thrombotic vascular diseases by investigating DNA length as a potential novel biomarker in cardiovascular disease. Methods Citrated plasma from healthy volunteers (n=363) and from ST-elevation myocardial infarction (MI) coronary aspirates (n=59) and peripheral arterial sites (n=96) were analyzed for DNA concentration, length, and nuclease activity. Total DNA concentration was measured with a fluorescent DNA binding dye, and elongated DNA assessed using qPCR and Agilent bioanalyzer high-sensitivity DNA chip. Nuclease activity was measured by single radial enzyme-diffusion (SRED) assay. Cardiac MRI assessed infarct size acutely and six months after the event. Results Coronary aspirates exhibited an elevation in total DNA, with a significant rise in high-molecular weight cell-free DNA in addition to an increase in sub-nucleosome fragmentation (Figure 1). Peripheral and coronary plasma samples from individuals with myocardial infarction (MI) showed elevated levels of total and elongated cfDNA. When compared to healthy individuals, the integrity of cfDNA was lower in peripheral samples and higher in coronary aspirates. During myocardial infarction, circulating nuclease activity was elevated systemically but low at the coronary culprit lesion site. Elongated high-molecular-weight cfDNA accumulates primarily in coronary vessels but is also elevated systemically. In coronary aspirates, nuclease activity negatively correlated with elongated DNA and DNA integrity (rs=0.6255, p<0.0001). Conclusions Elongated DNA accumulates during MI at the culprit lesion site. Its inverse correlation with nuclease activity underscores the role of nucleases in cleaving elongated DNA to shorter, soluble fragments. This study establishes DNA length and integrity as novel biomarkers in thrombotic cardiovascular disease.Figure 1:Coronary cfDNA lengthFigure 2:Nuclease activity and cfDNA

Red blood cell distribution width to albumin ratio as novel prognostic biomarker in cardiovascular disease

Abstract Background The red blood cell distribution width-to-albumin ratio (RAR) is a novel, cost-effective, and accessible biomarker for inflammation. Although numerous studies have investigated the relationship between RAR values and mortality in cardiovascular diseases (CVD), the generalizability and robustness of these individual studies' results remain uncertain, underscoring the necessity for a meta-analysis. Purpose To explore the pooled association between RAR and all-cause mortality in several CVDs. Methods A systematic search for eligible studies was conducted across scientific databases, including Scopus, Web of Science, PubMed, and ProQuest, up to February 1, 2024. Mortality hazard ratios (HRs) for the highest versus lowest RAR categories were pooled using the Generic Inverse Variance method. A dose-response meta-analysis was performed using generalized least-squares regression to estimate trends of HRs across dose categories. Publication bias was assessed by inspecting funnel plots and performing Egger's test. Subgroup analyses were based on the underlying CVDs. Statistical analyses were executed in RStudio software, with the significance threshold set at p < 0.05. Results This systematic review and meta-analysis reviewed 16 high-quality studies published between 2021 and 2024 with 30,933 participants. The underlying diseases in the included studies were Acute Myocardial Infarction (AMI), Heart Failure (HF), Atrial Fibrillation (AF), aortic valvular disease, stroke and pulmonary embolism. The lowest quartile range for RAR was 0.80–1.20 ml/g, while the highest quartile ranged from 6.09 to 15.45 ml/g. Pooled results with a random-effect model showed that patients with higher RAR had significantly increased risk of all-cause mortality (HR 1.88, 95%CI 1.59–2.23, p<0.0001, I² = 91%). Non-linearity was observed in the dose-response relationship; thus, it was primarily illustrated using the restricted cubic spline model, with the linear model provided for comparison. A 1 ml/g increase in RAR corresponded to a 27% rise in the predicted HR for all-cause mortality (HR 1.27, 95%CI 1.16–1.39; p < 0.0001). Subgroup analyses based on the primary CVD diagnoses in the included studies confirmed a significant association between RAR values and all-cause mortality across all subgroups (AMI: HR 2.43, 95%CI 1.52–3.87, I² = 45%; HF: HR 1.78, 95%CI 1.13–2.81, I² = 94%; Stroke: HR 1.58, 95%CI 0.94–2.67, I² = 90%; Other CVD: HR 1.97, 95%CI 1.34–2.89, I² = 57%; p for interaction 0.24). Conclusion This is the first meta-analysis demonstrating the positive dose-response association between RAR values and all-cause mortality in CVD. Thus, RAR may be considered as a simple and practical prognostic biomarker in CVD.Forest plot based on CVD diagnosisDose-response RAR value to HR

Awareness and perceptions of cardiologists towards systemic inflammation in atherosclerotic cardiovascular disease: A multi-national survey across 5 geographical regional sub-groups

Abstract Background Despite improvement in the management of atherosclerotic cardiovascular disease and chronic kidney disease (ASCVD+CKD), cardiovascular (CV) mortality remains unacceptably high. Evidence suggests that systemic inflammation (SI) is one of the key drivers for disease progression, yet clinical perceptions towards SI amongst cardiologists is poorly understood. Purpose FLAME-ASCVD (systemic inFLAMmation and rolE of hsCRP as a biomarker in AtheroSclerotic CardioVascular Disease) survey assessed cardiologist’s perceptions and awareness towards the role of SI and high sensitivity C-reactive protein (hsCRP) in patients (pts.) with ASCVD+CKD. Overall results were reported earlier1 and here we present results by sub-groups across 5 geographical regions. Methods This was a multinational, cross-sectional, online survey-based study conducted during March-May 2023. Eligible participants were cardiologists (interventional and general) who treated ≥15 ASCVD+CKD pts./month and practiced for ≥3 years. To minimize selection bias, specific study objective was not disclosed until eligibility criteria was met. An online questionnaire was used. Analysis included 5 sub-groups: Europe (EU: France, Germany, Italy, UK), Asia Pacific (AP: Australia, India), East Asia (EA: China, Japan), LatAm (LA: Brazil), Middle East (ME: Saudi Arabia). Descriptive statistics were used. Results Of the 1755 respondents, 602 eligible cardiologists (~60/country) completed the survey across the EU (n=241), AP (n=121), EA (n=120), LA (n=60), and ME (n=60). Hypertension, hyperlipidaemia, and lifestyle (diet/exercise) were consistently amongst the top 3-5 CV risk factors discussed with pts, across all regions. SI as a CV risk factor was discussed consistently less across regions (overall 10th at 43%, ³9th across regions, EU: 30%) (Table 1). Across regions, >70% of cardiologists acknowledged an intention to consider testing for SI, with the lowest being in the EU at 46%. Of those in the EU who did not consider testing, 53% (n=129) cited lack of medication as the reason. Across all regions, SI was consistently perceived as a risk factor for recurrent CV events (³65%) and residual inflammatory risk (47-85%), yet a noticeable proportion (52-92%) cardiologists acknowledged a need to learn more about SI in ASCVD+CKD (highest in ME, 92%). While overall (aided: 47% [n= 571]; unaided: 24%) considered hsCRP test for SI, few variations were observed (EA/AP/EU, aided: 55/52/33 and unaided: 22/24/03) (Table 2). Proven efficacy of hsCRP test, availability of medication for SI, and guideline recommendation were acknowledged as unmet needs to inform clinical care in ASCVD+CKD. Conclusion In this survey across regional sub-groups, discussion of SI as a CV risk factor was consistently low. Barring few noticeable variations in testing patterns in EU, perceptions towards SI were generally consistent. There is a need for better understanding and guidance on SI and hsCRP testing in practice.

Exploring protein biomarkers of physical activity and cardiovascular disease

Abstract Background The beneficial effects of physical activity on cardiovascular risk are well documented. Yet, the underlying mechanisms by which physical activity prevents cardiovascular disease remain unclear (1). Advancements of omics technology enables us to explore novel biomarkers of this complex molecular relationship and potentially unravel underlying molecular mechanisms involved in the progression of cardiovascular events. Purpose To investigate associations between long term physical activity and plasma proteins. As a second step investigate associations between identified plasma proteins and future myocardial infarction and mortality. Methods In the population based prospective cohort Uppsala Longitudinal Study of Adult Men (ULSAM), men were repeatedly investigated with a validated questionnaire on physical activity at ages 50, 60 and 70. 720 plasma proteins were analyzed at age 70 (n=782) with follow up on mortality data for 30 years. In the case-cohort MIMI (Markers of Imminent Myocardial Infarction study) the same plasma proteins were measured in disease-free individuals from six European cohorts. Individuals with acute myocardial infarction within 6 months from baseline were defined as cases (n=420) and up tto four cohort representatives per case were defined as controls (n=1598). Results Higher level of physical activity during 20 years was significantly associated with 12 plasma proteins, in a linear regression model adjusted for age, education and smoking after Bonferroni correction for multiple testing (p<0.000069). After additional adjustment for known cardiovascular risk factors (education, smoking status, BMI, blood pressure, low-density lipoprotein, diabetes diagnosis, and hypertensive-, lipid- or diabetes-treatment), all 12 proteins remained negatively associated with higher level of physical activity in the ULSAM cohort (p<0,05). Two proteins of the 12 proteins associated with physical activity were also significantly associated with future myocardial infarction in the MIMI study (FGF21 and IL6, p<0.05 for both) and 10 proteins were associated with increased mortality risk in the ULSAM cohort (p<0.05 for all). Moreover, proteins that were more negatively associated with physical activity were generally more positively associated with the risk of myocardial infarction in MIMI (Figure). Conclusion Multiple novel associations were found between long-term physical activity and plasma proteins. Some proteins were also associated with future myocardial infarction in an independent cohort which could suggest that the cardioprotective effects of physical activity to some degree may be mediated via the circulating proteome. Our findings encourage additional studies in order to understand the underlying causal mechanisms of these associations.

A direct comparison of non-invasive blood markers of liver fibrosis as associates of cardiovascular risk. The Athens cardiometabolic registry

Abstract Background Non-alcoholic fatty liver disease (NAFLD) is associated with increased risk of cardiovascular disease (CVD). This risk increases further with advanced liver fibrosis. To date, non-invasive tests (NIT) for liver fibrosis risk have not been established as diagnostic or prognostic biomarkers of CVD. Purpose The purpose of the study was to compare NIT of liver fibrosis with respect to their associations with vascular injury and cardiovascular (CV) events. Methods Individuals without clinically overt CVD and patients with CVD (total n=1,692), consecutively recruited in the Athens Cardiometabolic Registry, were analysed in this study. We retrospectively evaluated the association of NIT (i.e. Fibrosis-4 index (FIB-4), NAFLD Fibrosis score (NFS), BARD score) with indices of subclinical arterial injury (i.e. carotid wall thickness and atherosclerotic plaque for subclinical atherosclerosis, pulse wave velocity (PWV), for arterial stiffness) and the incidence of CV events. Patients were followed-up for CV events for a median time of 39 months. Results Elevated FIB-4 (>2.67) was associated with vascular damage [adjusted odds ratio (aOR)=2.97, 95% confidence intervals (CI)= 1.71-5.78 for maximal wall thickness (maxWT) and aOR=4.58, 95% CI=2.44-8.60 for PWV] across the entire study population, including those with or without clinically overt CVD. NFS> 0.675 showed similar associations but lacked significant correlation with arterial stiffness in coronary artery disease patients. FIB-4> 2.67 and BARD> 2 were associated with increased risk of the composite endpoint of CV death, acute myocardial infarction, or coronary revascularization [adjusted hazard ratio, (aHR)=2.00, 95% CI= 1.12-3.55 and aHR=3.36, 95% CI=1.03-11.0 for FIB-4 (n=842) and BARD (n=887) respectively]. FIB-4 and BARD as continuous variables confered incremental prognostic value compared to European Systematic Coronary Risk Evaluation 2 (SCORE2). Conclusion In a population with a wide range of cardiometabolic risk, FIB-4 was associated with both markers of atherosclerotic disease and arterial stiffness as well as with adverse CV events. These data support further investigation of FIB-4 as a biomarker of CV risk.

Inhibition of ceramide de novo synthesis mitigates atherosclerosis

Abstract Background Atherosclerosis is the primary underlying cause of cardiovascular disease. Plasma ceramides are positively correlated with an increased risk of major adverse cardiovascular events and serve as an alternative biomarker for cardiovascular disease. Importantly, ceramide levels are notably higher within atherosclerotic plaques. Therefore, inhibiting the ceramide de novo synthesis pathway in the plaques may serve as a potential therapeutic intervention for atherosclerosis. Purpose We aim to assess the therapeutic efficacy of a nano-formulation of myriocin, a targeted inhibitor of serine-palmitoyl transferase, the key enzyme in ceramide de novo synthesis, in treating atherosclerosis. Methods Myriocin was encapsulated within the hydrophobic core of lipid-based nanoparticles specifically designed for this study. Male and female ApoE-/- mice, subjected to an 8-week high-fat diet regimen, received either myriocin nanoparticles (treatment group) or empty nanoparticles (control group) intravenously once a week for 4 weeks at a dose of 1.3 mg/kg body weight while still on the high-fat diet. At the study endpoint, whole aortas were harvested and subjected to Oil Red O staining to quantify the total lesion area. Histological staining was performed on serial sections of the aortic root to analyze the cellular composition within the plaques. Plasma samples were collected for total cholesterol measurement and lipidomic analysis. Results Upon intravenous administration, myriocin nanoparticles exhibited preferential accumulation in atherosclerotic plaques. Compared to control mice, those treated with myriocin nanoparticles exhibited a 58.7% reduction in total lesion area in male ApoE-/- mice and a 64.6% reduction in female ApoE-/- mice (p<0.0001). Consistently, the lesion areas in the aortic root were also reduced in the treated mice. Notably, myriocin nanoparticles enhanced collagen and alpha-smooth muscle content, while reducing macrophages and neutral lipids within the plaques, leading to a reduced plaque vulnerability index and preventing rupture events. TUNEL staining revealed reduced cellular apoptosis in the atherosclerotic plaques, resulting in a smaller necrotic area in the treated mice. In vitro experiments demonstrated that myriocin nanoparticles effectively inhibited lipopolysaccharide-induced inflammatory response and oxidative stress in bone marrow-derived macrophages. Furthermore, myriocin nanoparticles restored the gene expression of enzymes involved in the fatty acid beta-oxidation pathway, such as ACOX1 and CPT1A, in foam cells. Conclusion Using myriocin nanoparticles to inhibit excessive ceramide production due to inflammation reduces inflammation, and ROS production, while restoring fatty acid beta-oxidation in macrophages. This approach emerges as an effective strategy for targeting ceramide de novo synthesis within atherosclerotic plaques, thereby mitigating atherosclerosis progression and improving plaque stability.

Incremental role of Cardio-Ankle Vascular Index in predicting ischemic threshold in patients with suspected coronary artery disease

Abstract Background The Cardio-Ankle Vascular Index (CAVI) quantifies arterial stiffness and has emerged as a potential biomarker for cardiovascular disease and its associated risk factors. This study explores the prognostic utility of CAVI in predicting myocardial ischemia in patients with suspected coronary artery disease (CAD). Additionally, it examines the relationship between global longitudinal strain (GLS) and the wall motion score index (WMSI) during both rest and stress echocardiography. Methods We conducted a retrospective analysis of patients with suspected CAD who underwent CAVI measurement and stress echocardiography within a three-month window. Coronary angiography was utilized to evaluate anatomical stenosis (>50%). We used multivariate analysis to identify predictors of the ischemic threshold. Results In our cohort of 127 patients (average age 61.4 ± 9.1, 78% male), the prevalence of hypertension, dyslipidemia, and diabetes was 76%, 68% and 47% of patients, respectively. Prior coronary intervention and a history of myocardial infarction were showed significant prevalence at 33% and 9%, respectively. CAD distribution included absence in 15%, left main (LM) disease in 8%, proximal left anterior descending (LAD) artery disease in 38%, mid-to-distal LAD disease in 50%, left circumflex artery disease in 46%, and right coronary artery disease in 57%. A robust correlation between GLS and WMSI was observed both at rest and during peak stress (r=0.68 and r=0.57, p<0.001, respectively). High CAVI (≥8.5), resting GLS in the LAD(<-15.5%), and peak GLS during stress (<-15.3%) emerged as significant ischemic threshold predictors in multivariate analysis (AUC = 0.82, p<0.0001). Conclusion The correlation between WMSI and GLS provides valuable insights into cardiac mechanics. These findings underscore the clinical relevance of incorporating CAVI, GLS, and WMSI into routine assessments for patients with suspected CAD to enhance prognostic stratification.ROC for final predictive model

Circulating levels of amyloid-beta (1-40) peptide associate with cardiometabolic traits and risk for metabolic dysfunction-associated steatotic liver disease

Abstract Background and Aims The ageing-associated amyloid-beta 1-40 peptide (Αbeta40) has recently emerged as a blood-based biomarker of cardiovascular disease (CVD). Its levels predict future cardiovascular death and MACE in patients with established CVD. The clinical value of Abeta40 in patients with metabolic syndrome who do not concomitantly have clinically overt CVD or other end-organ damage remains poorly understood. At the present study we investigated the clinical value of plasma Αbeta40 in patients with pre- or established metabolic syndrome who are at risk for metabolic dysfunction-associated steatotic liver disease (MASLD). Methods Αbeta40 was measured in plasma by enzyme-linked immunosorbent assay and cardiometabolic traits were determined in a mixed population (n=888) of patients with established (n=243) and pre- or without metabolic syndrome (n=645) who did not have clinical signs of established cardiometabolic disease (heart failure, coronary artery disease, MASLD, chronic kidney disease). The endpoint of the study was the association of Abeta40 with cardiometabolic indices and risk (BARD score) for MASLD. Odds ratio (OR) was calculated by ordinal regression analysis for the increasing incidence of metabolic syndrome components per increasing Abeta40 (1-SD) after adjustment for age, sex, smoking status and hyperlipidemia. Results Abeta40 levels are associated with the presence of metabolic syndrome and with higher odds for increasing incidence of metabolic syndrome components after adjustment for sex, age, smoking status, hypertension and dyslipidemia. Increased Abeta40 plasma levels were associated with decreased HDL-C levels (OR: 1.004, 95% CI:1.000-1.009, P=0.049) and increased triglyceride levels (OR: 1.007, 95%CI: 1.001-1.012, P=0.033) after adjustment for traditional cardiovascular risk factors (age, sex, smoking status, hypertension and hyperlipidemia). Among the total population (n=888), we identified n=473 subjects with increased MASLD risk if any of the following was present: hepatic steatosis index≥36 or fatty liver index≥60 or Fib-4 index≥2.67 or NAFLD score≥0.675. Among patients with high-risk for MASLD, patients with increased BARD score had significantly higher Abeta40 levels (median Abeta40 concentration in patients with BARD score≥2 vs. patients with BARD score<2: 54.9 pg/ml (IQR=38.25) vs. 41.9 pg/m (IQR=30.35), P=0.012). Increased Abeta40 levels were associated with increased odds for AST/ALT≥0.8 ratio and increased odds for BARD score≥2 after adjustment for age, sex, smoking status, hypertension and hyperlipidemia. Conclusion Our findings suggest that Abeta40 peptide is associated with cardiometabolic traits and risk for MASLD. Whether Abeta40 may emerge as an early cardiometabolic prognostic biomarker, future longitudinal studies are warranted to determine the prognostic value of Abeta40 for the development of cardiometabolic diseases in patients with metabolic syndrome.

Neutrophil-to-High-Density Lipoprotein Ratio (NHR) and Neutrophil-to-Lymphocyte Ratio (NLR) as prognostic biomarkers for incident cardiovascular disease and all-cause mortality: A comparison study

Cardiovascular diseases (CVD) remain a leading cause of global mortality, with atherosclerosis and inflammation playing pivotal roles in their development. The neutrophil-to-lymphocyte ratio (NLR) and neutrophil-to-HDL cholesterol ratio (NHR) have emerged as potential biomarkers for assessing CVD risk. In this community-based cohort study conducted in Taiwan, involving 3278 participants, we investigated the associations between NHR, NLR, and the risks of CVD and all-cause mortality. Our findings revealed that both NHR and NLR were effective in identifying individuals at high risk for CVD. However, when assessing their joint effect, NHR alone demonstrated a stronger predictive value for CVD prognosis than NLR or the combination of both markers. Furthermore, NLR alone showed potential as a predictor of all-cause mortality when compared with NHR alone or in combination with NLR and NHR. These findings underscore the complex interplay between inflammation and lipid metabolism in the pathogenesis of CVD. While NHR shows promise as a cost-effective tool for CVD risk assessment, NLR emerges potential as a prognostic marker for mortality. Further research is warranted to explore the dynamic changes in these markers and their implications for clinical practice.

Old, the new, more or less and the conundrum for biomarkers in cardiovascular diseases?

Biomarkers are critical for diagnostic, monitoring and management of cardiovascular diseases. Opportunities to improve how biomarkers are used are important. This editorial on biomarkers in coronary artery disease provides commentary on the study as well as broad benchmarks on what make ideal biomarkers. Biomarker discovery is difficult and opportunities to expand and improve their use should be encouraged.

An improved amperometric cholesterol biosensor based on cholesterol oxidase nanostructures for pre-diagnosis of myocardial infarction

Cholesterol plays a pivotal role in human health, serving as a crucial biomarker for cardiovascular diseases, including myocardial infarction. This study presents the development of an innovative amperometric cholesterol biosensor that enhances the detection and quantification of cholesterol levels in serum. The biosensor integrates cholesterol oxidase (ChOx) nanoparticles with a modified electrode, leveraging the unique properties of platinum nanoparticles (PtNPs) and graphene nanosheets (GNs) to improve sensitivity and stability. The synthesis of PtNPs was achieved using Camellia sinensis extract, while graphene oxide was reduced to form GNs. At 2.39 mg/mL or above is deemed a biomarker for cardiovascular disorders, peripheral artery disease, heart attack, diabetes mellitus, strokes, and hypertension. The monitoring of serum cholesterol level is therefore very significant. In the present study, an innovative amperometric cholesterol biosensor was constructed by immobilizing nanoparticles of cholesterol oxidase onto a pencil graphite (PG) electrode modified with graphene nanosheets (GNs), platinum nanoparticles (PtNPs), and chitosan (CHIT). At various stages of construction, the modified electrode was characterized by employing electrical impedance spectroscopy (EIS) and cyclic voltammetry (CV), scanning electron microscopy (SEM), and energy dispersive X-ray (EDX) spectroscopy. The biosensor responded best when run at 0.14Vs-1 at optimal pH and temperature of 8.0 and 35°C respectively. The biosensor has a wide linear range (0.1mg/mL-7.5 mg/mL), with great sensitivity (0.89 mA cm−1mgmL−1) and a low limit of detection (0.97 mg/mL). This research not only contributes to the field of biosensing but also offers a promising tool for the early diagnosis of cholesterol-related health issues, paving the way for enhanced cardiovascular disease management.

7669 Higher Serum Uric Acid Levels Are Associated With a Deleterious Metabolic and Inflammatory Profile in Brazilian Adolescents: An Exploratory Cross-Sectional Analysis

Abstract Disclosure: F.Q. Iorra: None. M.P. Guahnon: None. P.G. Rodrigues: None. A.T. Barcelos: None. F.V. Cureau: None. M. Drehmer: None. B.D. Schaan: None. Introduction: The association of high serum uric acid (sUA) with metabolic syndrome (MetS) and its components have been widely studied, but there are still gaps in how this breakdown product of purine metabolism might refine cardiovascular risk assessment and correlate with other cardiovascular disease biomarkers, especially in children and adolescents. Objective: To evaluate if sUA is associated with metabolic syndrome components and inflammatory markers in Brazilian adolescents. Methods: For this cross-sectional study, 786 individuals aged 12 to 17 years old, previously enrolled in the Study of Cardiovascular Risk in Adolescents (ERICA) - a Brazilian nationwide school based study, and who were from the city of Porto Alegre, had their sUA measured from blood samples that were stored in a biorepository. Anthropometric, biochemical and blood pressure data were collected. MetS diagnosis was based on the International Diabetes Federation age-specific criteria (2005), and insulin resistance was assessed by the HOMA-IR. In accordance with a non-parametric distribution, sUA was divided into quartiles (3.7 mg/dL, 4.4 mg/dL, 5.3 mg/dL for the 25th, 50th, and 75th percentiles, respectively) for the analysis. A linear regression model and a Poisson regression model with robust error variance were used, both adjusted for age, sex and skin color. Results: Participants were 14.8 ± 1.5 years-old, predominantly women (61.7%), self-identified as white skin color (72%) and eutrophic (66.9%). Through the linear regression model, significant positive associations were found between high sUA levels and body mass index Z-score, waist circumference, systolic and diastolic blood pressure, total cholesterol, triglycerides, HOMA-IR and C-reactive protein, and significant negative associations were observed for HDL cholesterol and adiponectin. No association was found between sUA and glucose or HbA1c. We also computed a metabolic risk score as the mean Z score of waist circumference, systolic blood pressure, HDL cholesterol (negative Z score), triglycerides and glucose, which was positive related to sUA (β ± SE = 0.13 ± 0.02; p < 0.001). Finally, the Poisson regression model was applied to estimate the prevalence ratios (PR) for MetS and its components, between adolescents with sUA in the 4th quartile to those in the 1st quartile. An association was observed for MetS (PR 6.37, 95% CI 1.39 - 29.21, p 0.017), high waist circumference (PR 4.61, 95% CI 2.55 - 8.36, p < 0.001) and increased triglycerides (PR 4.76, 95% CI 1.47 - 15.44, p 0.009). Conclusion: High sUA levels were associated with a worse metabolic and inflammatory profile, as well as a greater prevalence of MetS and some of its components in adolescents. Therefore, uric acid should be considered as a potential additional marker of cardiovascular risk in this population. Support: CNPq, CAPES, FIPE, FAPERGS. Presentation: 6/1/2024

9193 Higher Serum Uric Acid Levels Are Associated With a Deleterious Metabolic and Inflammatory Profile in Brazilian Adolescents: An Exploratory Cross-Sectional Analysis

Abstract Disclosure: F.Q. Iorra: None. M.P. Guahnon: None. P.G. Rodrigues: None. A.T. Barcelos: None. F.V. Cureau: None. M. Drehmer: None. B.D. Schaan: None. Introduction: The association of high serum uric acid (sUA) with metabolic syndrome (MetS) and its components have been widely studied, but there are still gaps in how this breakdown product of purine metabolism might refine cardiovascular risk assessment and correlate with other cardiovascular disease biomarkers, especially in children and adolescents. Objective: To evaluate if sUA is associated with metabolic syndrome components and inflammatory markers in Brazilian adolescents. Methods: For this cross-sectional study, 786 individuals aged 12 to 17 years old, previously enrolled in the Study of Cardiovascular Risk in Adolescents (ERICA) - a Brazilian nationwide school based study, and who were from the city of Porto Alegre, had their sUA measured from blood samples that were stored in a biorepository. Anthropometric, biochemical and blood pressure data were collected. MetS diagnosis was based on the International Diabetes Federation age-specific criteria (2005), and insulin resistance was assessed by the HOMA-IR. In accordance with a non-parametric distribution, sUA was divided into quartiles (3.7 mg/dL, 4.4 mg/dL, 5.3 mg/dL for the 25th, 50th, and 75th percentiles, respectively) for the analysis. A linear regression model and a Poisson regression model with robust error variance were used, both adjusted for age, sex and skin color. Results: Participants were 14.8 ± 1.5 years-old, predominantly women (61.7%), self-identified as white skin color (72%) and eutrophic (66.9%). Through the linear regression model, significant positive associations were found between high sUA levels and body mass index Z-score, waist circumference, systolic and diastolic blood pressure, total cholesterol, triglycerides, HOMA-IR and C-reactive protein, and significant negative associations were observed for HDL cholesterol and adiponectin. No association was found between sUA and glucose or HbA1c. We also computed a metabolic risk score as the mean Z score of waist circumference, systolic blood pressure, HDL cholesterol (negative Z score), triglycerides and glucose, which was positive related to sUA (β ± SE = 0.13 ± 0.02; p < 0.001). Finally, the Poisson regression model was applied to estimate the prevalence ratios (PR) for MetS and its components, between adolescents with sUA in the 4th quartile to those in the 1st quartile. An association was observed for MetS (PR 6.37, 95% CI 1.39 - 29.21, p 0.017), high waist circumference (PR 4.61, 95% CI 2.55 - 8.36, p < 0.001) and increased triglycerides (PR 4.76, 95% CI 1.47 - 15.44, p 0.009). Conclusion: High sUA levels were associated with a worse metabolic and inflammatory profile, as well as a greater prevalence of MetS and some of its components in adolescents. Therefore, uric acid should be considered as a potential additional marker of cardiovascular risk in this population. Support: CNPq, CAPES, FIPE, FAPERGS. Presentation: 6/1/2024

Non-enzymatic cholesterol biosensor: Electrochemical sensing based on peptide-polylactic acid thin film

Cholesterol is a fundamental lipid prevalent in eukaryotic cell membranes and circulating in the bloodstream bound to lipoproteins. It serves as a precursor to steroid hormones and is regarded as a biomarker for cardiovascular disease and other metabolic disorders. Numerous cholesterol detection methods predominantly rely on enzymes, which suffer from instability, leading to non-cost-effective biosensors with low sensitivity and poor reusability. Therefore, monitoring cholesterol levels with a feasible, rapid, and stable biosensor is critical for diagnosing and treating various disorders. This study aimed to develop a non-enzymatic cholesterol biosensor based on a selected cholesterol recognition peptide as the detection element. Screen-printed carbon electrodes (SPEs) modified with biocompatible poly-L-lactic acid (PLLA) porous nanomembranes (NMs) were utilized as support for the covalent immobilization of the peptide. Data obtained from electrochemical impedance spectroscopy (EIS) demonstrated the peptide's effective binding affinity towards cholesterol, paving the way for its implementation. The determination of cholesterol with the proposed biosensor exhibited a low limit of detection of 6.31 μM with linear responses ranging from 2–15 μM and 20–40 μM. These findings present an alternative method for cholesterol sensing by integrating novel peptides as biorecognition motifs with biocompatible polymeric materials, potentially useful as biocompatible and future point-of-care sensors.

MicroRNAs as diagnostic biomarkers in cardiovascular diseases that cause sudden cardiac death: comparison between methods used in discovery and validation phases in major research studies

MicroRNAs as diagnostic biomarkers in cardiovascular diseases that cause sudden cardiac death: comparison between methods used in discovery and validation phases in major research studies

Effect of nuts on lipid profile and inflammatory biomarkers in atherosclerotic cardiovascular disease: a systematic review and meta-analysis of randomized controlled trials.

Nut-enriched diets are related to improve lipid and inflammatory biomarkers in meta-analyses in the context of primary cardiovascular prevention. However, primary studies on secondary cardiovascular prevention are scarce and controversial. This systematic review and meta-analysis aimed to evaluate the effect of nut supplementation on lipid and inflammatory profiles in individuals with atherosclerotic cardiovascular disease, and the frequency of adverse events. Six databases were used for research: PubMed, EMBASE, BVS, Cochrane Library, Web of Science, and ClinicalTrials.gov, until February 2023, with no language restrictions. We performed random-effects meta-analyses to compare nut-enriched diets vs. control diets for pre-post intervention changes. The Grading of Recommendations Assessment, Development and Evaluation (GRADE) system assessed the evidence's certainty. From the 5187 records identified, eight publications containing data referring to five randomized clinical trials involving 439 participants were included in the final analyses. The nuts evaluated were almonds, pecans, Brazil nuts, and mixed nuts, with doses ranging between 5g and 85g (median: 30g/day). The intervention time varied between 6 and 12 weeks. Compared to nut-free diets, nut intake did not have a statistically significant effect on lipid profile biomarkers, except on the atherogenic index (MD: -0.32 [95% CI -0.58 to -0.06], I2 = 0% - moderate certainty of the evidence). Similarly, there was no effect of nuts on inflammatory profile biomarkers. It was not possible to aggregate data on adverse events. Nut supplementation did not change lipid and inflammatory profiles in the secondary cardiovascular prevention setting.

Synergy Between NK Cells and Monocytes in Potentiating Cardiovascular Disease Risk in Severe COVID-19.

Evidence suggests that COVID-19 predisposes to cardiovascular diseases (CVDs). While monocytes/macrophages play a central role in the immunopathogenesis of atherosclerosis, less is known about their immunopathogenic mechanisms that lead to CVDs during COVID-19. Natural killer (NK) cells, which play an intermediary role during pathologies like atherosclerosis, are dysregulated during COVID-19. Here, we sought to investigate altered immune cells and their associations with CVD risk during severe COVID-19. We measured plasma biomarkers of CVDs and determined phenotypes of circulating immune subsets using spectral flow cytometry. We compared these between patients with severe COVID-19 (severe, n=31), those who recovered from severe COVID-19 (recovered, n=29), and SARS-CoV-2-uninfected controls (controls, n=17). In vivo observations were supported using in vitro assays to highlight possible mechanistic links between dysregulated immune subsets and biomarkers during and after COVID-19. We performed multidimensional analyses of published single-cell transcriptome data of monocytes and NK cells during severe COVID-19 to substantiate in vivo findings. During severe COVID-19, we observed alterations in cardiometabolic biomarkers including oxidized-low-density lipoprotein, which showed decreased levels in severe and recovered groups. Severe patients exhibited dysregulated monocyte subsets, including increased frequencies of proinflammatory intermediate monocytes (also observed in the recovered) and decreased nonclassical monocytes. All identified NK-cell subsets in the severe COVID-19 group displayed increased expression of activation and tissue-resident markers, such as CD69 (cluster of differentiation 69). We observed significant correlations between altered immune subsets and plasma oxidized-low-density lipoprotein levels. In vitro assays revealed increased uptake of oxidized-low-density lipoprotein into monocyte-derived macrophages in the presence of NK cells activated by plasma of patients with severe COVID-19. Transcriptome analyses confirmed enriched proinflammatory responses and lipid dysregulation associated with epigenetic modifications in monocytes and NK cells during severe COVID-19. Our study provides new insights into the involvement of monocytes and NK cells in the increased CVD risk observed during and after COVID-19.