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A special issue of Diagnostics (ISSN 2075-4418). This special issue belongs to the section "Pathology and Molecular Diagnostics".

Deadline for manuscript submissions: closed (31 December 2020) | Viewed by 56808

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Prof. Dr. Paolo Zamboni

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Vascular Diseases Center, University of Ferrara, Via Aldo Moro 8, 44124 Ferrara, Italy
Interests: chronic venous disorders; venous leg ulcers; neurovascular disease; genetic polymorphism; hemodynamics; cerebral circulation; CCSVI; ultrasound; exercise; PAD
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Dear Colleagues,

The arterial, venous and lymphatic components of the human circulation are a fascinating field of research. The physical parameters of blood flow, blood oxygenation, imaging patterns, electric signaling and, of course, circulating and wall molecules are all potential biomarkers to improve our understanding and treatment of vascular disease. The latter is a heterogenous field of medicine, linked with metabolic and inflammatory disorders. In addition, there is a complex cross talk between circulating cells, the coagulation cascade, endothelial cells, and inflammatory cytokines that needs to be further elucidated. Finally, vascular and endovascular surgery, pharmacological therapy, cell therapy and rehabilitative treatments urgently need predictive biomarkers to address indications and monitor the outcome of interventions during follow-up.

Prof. Dr. Paolo Zamboni
Guest Editor

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Keywords

biomarkers
blood flow
vascular pharmacology
tissue oxygenation
electric biosignaling
imaging techniques
ultrasound
inflammation
vascular–endovascular surgery
wound healing
cell therapy
exercise
vascular rehabilitation
metabolism
sensors
coagulation

Published Papers (12 papers)

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Editorial

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4 pages, 432 KiB

Open AccessEditorial

Vascular Biomarkers: Physics Parameters and Circulating Molecules Can Be Two Faces of the Same Coin

by Paolo Zamboni

Diagnostics 2021, 11(2), 217; https://doi.org/10.3390/diagnostics11020217 - 2 Feb 2021

Viewed by 2151

Abstract

The arterial, venous and lymphatic conduits of human circulation are a fascinating field of research [...] Full article

(This article belongs to the Special Issue Biomarkers of Vascular Diseases)

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10 pages, 4261 KiB

Open AccessEditorial

COVID-19 as a Vascular Disease: Lesson Learned from Imaging and Blood Biomarkers

by Paolo Zamboni

Diagnostics 2020, 10(7), 440; https://doi.org/10.3390/diagnostics10070440 - 29 Jun 2020

Cited by 16 | Viewed by 14858

Abstract

COVID-19, a disease initially thought to be prominently an interstitial pneumonia with varying degrees of severity, can be considered a vascular disease with regards to serious complications and causes of mortality. Quite recently, blood clots have emerged as the common factor unifying many of the symptoms initially attributed without an explanation to COVID-19. Cardiovascular biomarkers and particularly, D-dimer and troponin appear to be very powerful prognostic markers, signaling the need for earlier and more aggressive interventions and treatments in order to avoid and/or minimize arterial/venous thromboembolism and myocardial infarct. The ultrasound imaging patterns at both the lung and peripheral vascular level can also be very useful weapons that have the advantage of being able to monitor longitudinally the clinical picture, something that real-time PCR/nasopharyngeal swab is not able to do and that CT can only pursue with significant radiation exposure. A lesson learned in the early phase of the COVID-19 pandemic suggests quitting and starting again with targeted imaging and blood vascular biomarkers. Full article

(This article belongs to the Special Issue Biomarkers of Vascular Diseases)

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Research

Jump to: Editorial, Review

15 pages, 33838 KiB

Open AccessArticle

Doppler Ultrasound Monitoring of Echogenicity in Asymptomatic Subcritical Carotid Stenosis and Assessment of Response to Oral Supplementation of Vitamin K2 (PLAK2 Randomized Controlled Trial)

by Yamume Tshomba, Domenico Baccellieri, Niccolò Carta, Giuseppe Cilli, Vincenzo Ardita, Luca Apruzzi, Diletta Loschi, Andrea Kahlberg, Luca Bertoglio, Renata Castellano, Elisa Simonini, Felicita Andreotti and Roberto Chiesa

Diagnostics 2021, 11(2), 229; https://doi.org/10.3390/diagnostics11020229 - 3 Feb 2021

Cited by 2 | Viewed by 2639

Abstract

Background: Plaque composition may predict the evolution of carotid artery stenosis rather than its sole extent. The grey scale median (GSM) value is a reproducible and standardized value to report plaque echogenicity as an indirect measure of its composition. We monitored plaque composition in asymptomatic subcritical carotid stenosis and evaluated the effect of an oral modulating calcification factor (vitamin K2). Methods: Carotid plaque composition was assessed by GSM value. Monitoring the effects of standard therapy (acetylsalicylic acid and low–medium dosage statin) (acetylsalicylic acid (ASA) arm) or standard therapy plus vitamins K2 oral supplementation (ASA + K2 arm) over a 12 months period was conducted using an ultrasound scan in a prospective, open-label, randomized controlled trial (PLAK2). Results: Sixty patients on low–medium dosage statin therapy were enrolled and randomized (30 per arm) to either ASA + K2 or ASA alone. Thirty-seven patients (61.6%) showed at 12 months a stable plaque with a mean increase in the GSM value in respect to the baseline of 2.6% with no differences between the two study arms (p = 0.66). Fifteen patients (25%) showed an 8% GSM value reduction respect the baseline with no differences between the two study arms (p = 0.99). At multivariable analysis, the adjusted mean (95% confidence interval) GSM change per month from baseline was greater in the ASA + K2 arm (−0.55 points, p = 0.048) compared to ASA alone (−0.18 points, p = 0.529). Conclusions: Carotid plaque composition monitoring through GSM value represents a laborious procedure. Although its use may not be applied to everyday practice, a specific application consists in evaluating the effect of pharmacological therapy on plaque composition. This 12 months randomized trial showed that the majority of subcritical asymptomatic carotid plaque on treatment with low–medium dosage statin presented a stable or increased echogenicity. Although vitamin K2 beyond standard therapy did not determine a significant change in plaque composition, for those who presented with GSM reduction it did enhance a GSM monthly decline. Full article

(This article belongs to the Special Issue Biomarkers of Vascular Diseases)

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11 pages, 4276 KiB

Open AccessArticle

Consistent Cerebral Blood Flow Covariance Networks across Healthy Individuals and Their Similarity with Resting State Networks and Vascular Territories

by Alice Pirastru, Laura Pelizzari, Niels Bergsland, Marta Cazzoli, Pietro Cecconi, Francesca Baglio and Maria Marcella Laganà

Diagnostics 2020, 10(11), 963; https://doi.org/10.3390/diagnostics10110963 - 17 Nov 2020

Cited by 5 | Viewed by 2054

Abstract

Cerebral blood flow (CBF) represents the local blood supply to the brain, and it can be considered a proxy for neuronal activation. Independent component analysis (ICA) can be applied to CBF maps to derive patterns of spatial covariance across subjects. In the present study, we aimed to assess the consistency of the independent components derived from CBF maps (CBF-ICs) across a cohort of 92 healthy individuals. Moreover, we evaluated the spatial similarity of CBF-ICs with respect to resting state networks (RSNs) and vascular territories (VTs). The data were acquired on a 1.5 T scanner using arterial spin labeling (ASL) and resting state functional magnetic resonance imaging. Similarity was assessed considering the entire ASL dataset. Consistency was evaluated by splitting the dataset into subsamples according to three different criteria: (1) random split of age and sex-matched subjects, (2) elderly vs. young, and (3) males vs. females. After standard preprocessing, ICA was performed. Both consistency and similarity were assessed by visually comparing the CBF-ICs. Then, the degree of spatial overlap was quantified with Dice Similarity Coefficient (DSC). Frontal, left, and right occipital, cerebellar, and thalamic CBF-ICs were consistently identified among the subsamples, independently of age and sex, with fair to moderate overlap (0.2 < DSC ≤ 0.6). These regions are functional hubs, and their involvement in many neurodegenerative pathologies has been observed. As slight to moderate overlap (0.2< DSC < 0.5) was observed between CBF-ICs and some RSNs and VTs, CBF-ICs may mirror a combination of both functional and vascular brain properties. Full article

(This article belongs to the Special Issue Biomarkers of Vascular Diseases)

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12 pages, 2302 KiB

Open AccessArticle

Longitudinal Magnetic Resonance Imaging of Cerebral Microbleeds in Multiple Sclerosis Patients

by Karthikeyan Subramanian, David Utriainen, Deepa P. Ramasamy, Sean K. Sethi, Ferdinand Schweser, John Beaver, Jesper Hagemeier, Bianca Weinstock-Guttman, Rajasimhan Rajagovindan, Robert Zivadinov and Ewart Mark Haacke

Diagnostics 2020, 10(11), 942; https://doi.org/10.3390/diagnostics10110942 - 12 Nov 2020

Cited by 5 | Viewed by 3474

Abstract

We hypothesized that cerebral microbleeds (CMBs) in multiple sclerosis (MS) patients will be detected with higher prevalence compared to healthy controls (HC) and that quantitative susceptibility mapping (QSM) will help remove false positives seen in susceptibility weighted imaging (SWI). A cohort of 100 relapsing remitting MS subjects scanned at 3T were used to validate a set of CMB detection guidelines specifically using QSM. A second longitudinal cohort of 112 MS and 25 HCs, also acquired at 3T, was reviewed across two time points. Both cohorts were imaged with SWI and fluid attenuated inversion recovery. Fourteen subjects in the first cohort (14%, 95% CI 8–21%) and twenty-one subjects in the second cohort (18.7%, 95% CI 11–27%) had at least one CMB. The combined information from SWI and QSM allowed us to discern stable CMBs and new CMBs from potential mimics and evaluate changes over time. The longitudinal results demonstrated that longer disease duration increased the chance to develop new CMBs. Higher age was also associated with increased CMB prevalence for MS and HC. We observed that MS subjects developed new CMBs between time points, indicating the need for longitudinal quantitative imaging of CMBs. Full article

(This article belongs to the Special Issue Biomarkers of Vascular Diseases)

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12 pages, 2933 KiB

Open AccessArticle

The Effects of Thermal Water Physical Exercise in Patients with Lower Limb Chronic Venous Insufficiency Monitored by Bioimpedance Analysis

by Erica Menegatti, Anselmo Pagani, Giampiero Avruscio, Marianna Mucignat and Sergio Gianesini

Diagnostics 2020, 10(11), 889; https://doi.org/10.3390/diagnostics10110889 - 31 Oct 2020

Cited by 5 | Viewed by 2599

Abstract

Background: Lower limb chronic venous diseases (CVD) lead to possible edema. The aim of the present investigation was to study the effect of thermal aquatic standardize exercise on lower limb edema control in CVD patients assessed by bioimpedance analysis (BIA). Methods: Seventeen patients (34 legs) affected by CVD clinical class CEAP C3, 4c, Ep, As, Pr were included. All the cohort performed a standardized exercise protocol in thermal water environment for a total of five sessions. BIA, leg volume, and heart rate at rest were measured. Results: After the five exercise sessions, BIA showed a significant percentage of extracellular water (ECW) reduction from 42.1 ± 5.8 to 41.24 ± 5.5%; p < 0.001. Moreover, an improvement of resistance (p < 0.0009) and reactance (p < 0.009) was assessed. At the same time, the leg volume reduction rate was 15.7%, p <0.0001. A moderate-strong correlation was found between % ECW and leg volume variation (R = 0.59, p < 0.01). Finally, a significant HR at rest reduction was recorded, p < 0.0001. Conclusion: The investigated exercise protocol significantly affects the lower limb volume, and BIA parameters related to the tissue drainage improvement. The correlation founded between the ECW rate and volume variations suggest the possible use of BIA as a biomarker for monitoring the treatments aimed to reduce edema in CVD. Full article

(This article belongs to the Special Issue Biomarkers of Vascular Diseases)

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15 pages, 1104 KiB

Open AccessArticle

Rehabilitation Improves Mitochondrial Energetics in Progressive Multiple Sclerosis: The Significant Role of Robot-Assisted Gait Training and of the Personalized Intensity

by Fabio Manfredini, Sofia Straudi, Nicola Lamberti, Simone Patergnani, Veronica Tisato, Paola Secchiero, Francesco Bernardi, Nicole Ziliotto, Giovanna Marchetti, Nino Basaglia, Massimo Bonora and Paolo Pinton

Diagnostics 2020, 10(10), 834; https://doi.org/10.3390/diagnostics10100834 - 17 Oct 2020

Cited by 15 | Viewed by 3322

Abstract

Abnormal levels of pyruvate and lactate were reported in multiple sclerosis (MS). We studied the response of markers of mitochondrial function to rehabilitation in relation to type, intensity and endurance performance in severely disabled MS patients. Forty-six progressive MS patients were randomized to receive 12 walking sessions of robot-assisted gait training (RAGT, n = 23) or conventional overground therapy (CT, n = 23). Ten healthy subjects were also studied. Blood samples were collected to determine lactate, pyruvate, and glutathione levels and lactate/pyruvate ratio pre–post rehabilitation. In vivo muscle metabolism and endurance walking capacity were assessed by resting muscle oxygen consumption (rmVO₂) using near-infrared spectroscopy and by six-minute walking distance (6MWD), respectively. The levels of mitochondrial biomarkers and rmVO₂, altered at baseline with respect to healthy subjects, improved after rehabilitation in the whole population. In the two groups, an enhanced response was observed after RAGT compared to CT for lactate (p = 0.012), glutathione (<0.001), lactate/pyruvate ratio (p = 0.08) and rmVO₂ (p = 0.07). Metabolic biomarkers and 6MWD improvements were exclusively correlated with a training speed markedly below individual gait speed. In severely disabled MS patients, rehabilitation rebalanced altered serum metabolic and muscle parameters, with RAGT being more effective than CT. A determinable slow training speed was associated with better metabolic and functional recovery. Trial Registration: ClinicalTrials.gov NCT02421731. Full article

(This article belongs to the Special Issue Biomarkers of Vascular Diseases)

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15 pages, 4511 KiB

Open AccessArticle

Association of Major Histocompatibility Complex Class I Related Chain A/B Positive Microparticles with Acute Myocardial Infarction and Disease Severity

by Songpol Haohan, Burabha Pussadhamma, Amonrat Jumnainsong, Wit Leuangwatthananon, Pattarapong Makarawate, Chanvit Leelayuwat and Nantarat Komanasin

Diagnostics 2020, 10(10), 766; https://doi.org/10.3390/diagnostics10100766 - 29 Sep 2020

Cited by 1 | Viewed by 2172

Abstract

Background: Various cell types undergo activation and stress during atherosclerosis resulting in the development of acute myocardial infarction (AMI) in coronary artery disease (CAD). Major histocompatibility complex class I related chain A and B (MICA/B) can be expressed on the surface of activated and stressed cells and released into blood circulation in several forms including microparticles (MICA/B⁺ MPs) from various cell types. We aimed to investigate the association of these MICA/B⁺ MPs with the presence of AMI. Fifty-one AMI and 46 age-matched control subjects were recruited. Methods: Levels of MICA/B⁺ MPs derived from various parent cells including endothelial cells, platelets, monocytes, neutrophils, and T lymphocytes were determined by flow cytometry. Results: The levels and proportion of MICA/B⁺ MPs from all types of cell origin were significantly increased in AMI patients compared to those of the controls. A multivariate regression model showed an independent association between MICA/B⁺ MPs and AMI (OR = 11.6; 95% CI = 2.8, 47.3). Interestingly, based on the disease severity, we found that the levels of MICA/B⁺ MPs were significantly elevated in the ST-segment elevation myocardial infarction (STEMI) compared to the non-STEMI (NSTEMI) patients. Moreover, an independent association of MICA/B⁺ MPs with the occurrence of STEMI was also demonstrated (OR = 4.1; 95% CI = 1.5, 16.7). Conclusions: These results suggest that MICA/B⁺ MPs are associated with AMI and disease severity. They may act as mediators contributing to the pathological process of AMI. Alternatively, they are the results of various cell activations contributing to AMI. Full article

(This article belongs to the Special Issue Biomarkers of Vascular Diseases)

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15 pages, 1408 KiB

Open AccessArticle

Biomarker Utility for Peripheral Artery Disease Diagnosis in Real Clinical Practice: A Prospective Study

by Alexandr Ceasovschih, Victorita Sorodoc, Viviana Onofrei (Aursulesei), Dan Tesloianu, Cristina Tuchilus, Ecaterina Anisie, Antoniu Petris, Cristian Statescu, Elisabeta Jaba, Alexandra Stoica, Elena-Daniela Grigorescu, Irina M. Jaba and Laurentiu Sorodoc

Diagnostics 2020, 10(9), 723; https://doi.org/10.3390/diagnostics10090723 - 20 Sep 2020

Cited by 18 | Viewed by 5219

Abstract

Peripheral arterial disease (PAD) is a common manifestation of generalized atherosclerosis, which affects more than 200 million patients worldwide. Currently, there is no ideal biomarker for PAD risk stratification and diagnosis. The goal of this research was to investigate the levels of inflammation biomarkers and cystatin C and to explore their utility for the diagnosis of PAD. The study included 296 participants, distributed in two groups: 216 patients diagnosed with PAD and 80 patients without PAD as controls. All studied biomarker levels (C-reactive protein, CRP; fibrinogen; erythrocyte sedimentation rate, ESR; neopterin; beta 2-microglobulin, B2-MG; and cystatin C) were significantly higher in the PAD group and indirectly correlated with the ankle–brachial index (ABI). The final logistic regression model included an association of neopterin, fibrinogen, and cystatin C as the most efficient markers for the prediction of PAD diagnosis. When comparing the area under the curve (AUC) for all biomarkers, the value for neopterin was significantly higher than those of all the other analyzed biomarkers. In agreement with previous studies, this research shows that markers such as fibrinogen, CRP, ESR, B2-MG, and cystatin C have significant value for the diagnosis of PAD, and also clearly underlines the accuracy of neopterin as a leading biomarker in PAD prediction. Full article

(This article belongs to the Special Issue Biomarkers of Vascular Diseases)

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10 pages, 571 KiB

Open AccessArticle

Serum Neurofilament Light Chain Levels are Associated with Lower Thalamic Perfusion in Multiple Sclerosis

by Dejan Jakimovski, Niels Bergsland, Michael G. Dwyer, Deepa P. Ramasamy, Murali Ramanathan, Bianca Weinstock-Guttman and Robert Zivadinov

Diagnostics 2020, 10(9), 685; https://doi.org/10.3390/diagnostics10090685 - 11 Sep 2020

Cited by 5 | Viewed by 3877

Abstract

Both perfusion-weighted imaging (PWI) measures and serum neurofilament light (sNfL) chain levels have been independently associated with disability in multiple sclerosis (MS) patients. This study aimed to determine whether these measures are correlated to each other or independently describe different MS processes. For this purpose, 3T MRI dynamic susceptibility contrast (DSC)–PWI and single-molecule assay (Simoa)-based sNfL methods were utilized when investigating 86 MS patients. The perfusion measures of mean transit time (MTT), cerebral blood volume (CBV), and cerebral blood flow (CBF) were derived for the normal-appearing whole brain (NAWB), the normal-appearing white matter (NAWM), the gray matter (GM), the deep GM (DGM), and the thalamus. The normalized CBV and CBF (nCBV and nCBV) were calculated by dividing by the corresponding NAWM measure. Age- and sex-adjusted linear regression models were used to determine associations between the DSC–PWI and sNfL results. False discovery rate (FDR)-adjusted p-values < 0.05 were considered statistically significant. A greater age and thalamic MTT were independently associated with higher sNfL levels (p < 0.001 and p = 0.011) and explained 36.9% of sNfL level variance. NAWM MTT association with sNfL levels did not survive the FDR correction. In similar models, a lower thalamic nCBF and nCBV were both associated with greater sNfL levels (p < 0.001 and p = 0.022), explaining 37.8% and 44.7% of the variance, respectively. In conclusion, higher sNfL levels were associated with lower thalamic perfusion. Full article

(This article belongs to the Special Issue Biomarkers of Vascular Diseases)

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16 pages, 8426 KiB

Open AccessArticle

Preliminary Post-Mortem COVID-19 Evidence of Endothelial Injury and Factor VIII Hyperexpression

by Luigi Cipolloni, Francesco Sessa, Giuseppe Bertozzi, Benedetta Baldari, Santina Cantatore, Roberto Testi, Stefano D’Errico, Giulio Di Mizio, Alessio Asmundo, Sergio Castorina, Monica Salerno and Cristoforo Pomara

Diagnostics 2020, 10(8), 575; https://doi.org/10.3390/diagnostics10080575 - 9 Aug 2020

Cited by 49 | Viewed by 7078

Abstract

(1) Background: The current outbreak of COVID-19 infection is an ongoing challenge and a major threat to public health that requires surveillance, prompt diagnosis, as well as research efforts to understand the viral pathogenesis. Despite this, to date, very few studies have been performed concerning autoptic specimens. Therefore, this study aimed: (i) to reiterate the importance of the autoptic examination, the only method able to precisely define the cause of death; (ii) to provide a complete post-mortem histological and immunohistochemical investigation pattern capable of diagnosing death from COVID-19 infection. (2) Methods: In this paper, the lung examination of two subjects who died from COVID-19 are discussed, comparing the obtained data with those of the control, a newborn who died from pneumonia in the same pandemic period. (3) Results: The results of the present study suggest that COVID-19 infection can cause different forms of acute respiratory distress syndrome (ARDS), due to diffuse alveolar damage and diffuse endothelial damage. Nevertheless, different patterns of cellular and cytokine expression are associated with anti-COVID-19 antibody positivity, compared to the control case. Moreover, in both case studies, it is interesting to note that COVID-19, ACE2 and FVIII positivity was detected in the same fields. (4) Conclusions: COVID-19 infection has been initially classified as exclusively interstitial pneumonia with varying degrees of severity. Subsequently, vascular biomarkers showed that it can also be considered a vascular disease. The data on Factor VIII discussed in this paper, although preliminary and limited in number, seem to suggest that the thrombogenicity of Sars-CoV2 infection might be linked to widespread endothelial damage. In this way, it would be very important to investigate the pro-coagulative substrate both in all subjects who died and in COVID-19 survivors. This is because it may be hypothesized that the different patterns with which the pathology is expressed could depend on different individual susceptibility to infection or a different personal genetic-clinical background. In light of these findings, it would be important to perform more post-mortem investigations in order to clarify all aspects of the vascular hypothesis in the COVID-19 infection. Full article

(This article belongs to the Special Issue Biomarkers of Vascular Diseases)

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Review

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27 pages, 525 KiB

Open AccessReview

A Systematic Review of Cerebral Functional Near-Infrared Spectroscopy in Chronic Neurological Diseases—Actual Applications and Future Perspectives

by Augusto Bonilauri, Francesca Sangiuliano Intra, Luigi Pugnetti, Giuseppe Baselli and Francesca Baglio

Diagnostics 2020, 10(8), 581; https://doi.org/10.3390/diagnostics10080581 - 12 Aug 2020

Cited by 30 | Viewed by 5027

Abstract

Background: The management of people affected by age-related neurological disorders requires the adoption of targeted and cost-effective interventions to cope with chronicity. Therapy adaptation and rehabilitation represent major targets requiring long-term follow-up of neurodegeneration or, conversely, the promotion of neuroplasticity mechanisms. However, affordable and reliable neurophysiological correlates of cerebral activity to be used throughout treatment stages are often lacking. The aim of this systematic review is to highlight actual applications of functional Near-Infrared Spectroscopy (fNIRS) as a versatile optical neuroimaging technology for investigating cortical hemodynamic activity in the most common chronic neurological conditions. Methods: We reviewed studies investigating fNIRS applications in Parkinson’s Disease (PD), Alzheimer’s Disease (AD) and Mild Cognitive Impairment (MCI) as those focusing on motor and cognitive impairment in ageing and Multiple Sclerosis (MS) as the most common chronic neurological disease in young adults. The literature search was conducted on NCBI PubMed and Web of Science databases by PRISMA guidelines. Results: We identified a total of 63 peer-reviewed articles. The AD spectrum is the most investigated pathology with 40 articles ranging from the traditional monitoring of tissue oxygenation to the analysis of functional resting-state conditions or cognitive functions by means of memory and verbal fluency tasks. Conversely, applications in PD (12 articles) and MS (11 articles) are mainly focused on the characterization of motor functions and their association with dual-task conditions. The most investigated cortical area is the prefrontal cortex, since reported to play an important role in age-related compensatory mechanism and neurofunctional changes associated to these chronic neurological conditions. Interestingly, only 9 articles applied a longitudinal approach. Conclusion: The results indicate that fNIRS is mainly employed for the cross-sectional characterization of the clinical phenotypes of these pathologies, whereas data on its utility for longitudinal monitoring as surrogate biomarkers of disease progression and rehabilitation effects are promising but still lacking. Full article

(This article belongs to the Special Issue Biomarkers of Vascular Diseases)

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