Markers Of Thrombosis Research Articles

ОСОБЕННОСТИ ЛАБОРАТОРНЫХ ПОКАЗАТЕЛЕЙ ВОСПАЛЕНИЯ И ТРОМБОЗА У ПАЦИЕНТОВ С НОВОЙ КОРОНАВИРУСНОЙ ИНФЕКЦИЕЙ SARS-COV-2 И ТРОМБОЭМБОЛИЕЙ ЛЕГОЧНОЙ АРТЕРИИ

Introduction. The COVID-19 pandemic continues with over 600 million cases and over 6 million deaths worldwide according to WHO. The state of hypercoagulation is a key feature of the course of COVID-19 which often leads to the development of serious cardiovascular events and adverse outcomes. There is a higher risk of all-cause mortality in the COVID-19 cohort with thrombotic complications, and mortality among patients with COVID-19 and PE is significantly higher than in patients with either condition alone, indicating a life-threatening additive effect of the combination of COVID-19 and PE. Thus, it is necessary to study further the features of inflammation and thrombosis parameters in patients with COVID-19 given the high prevalence of thrombotic complications among this group of patients. Objective. To define features of inflammation and thrombosis laboratory markers in patients with COVID-19 and pulmonary embolism. Materials and Methods. The study included n = 116 patients with COVID-19 hospitalized in 4-th city clinical hospital of Minsk named after N.E.Sauchenko in whom a thrombotic event – pulmonary embolism (PE) – was developed. The mean age of the patients was 64.7 ± 11.3 years, with 53 (45.7%) male and 63 (54.3%) female patients, respectively. The study group consisted of patients with COVID-19 and confirmed diagnosis of PE (n = 37) and the comparison group consisted of patients with COVID-19 without PE (n = 79). Patients in the groups were comparable by sex, age, presence of traditional risk factors, and COVID-19 severity. The parameters associated with thrombosis were analyzed in general blood count, hemostasiogram, biochemical blood analysis at the moment of confirmation or exclusion of PE using computer tomographic angiography of the pulmonary arteries. Results. Intergroup comparison of laboratory parameters in the group of patients with COVID-19 and confirmed diagnosis of PE in comparison with the group of patients with COVID-19 without PE showed a significantly higher mean group leukocyte count: 10,59 (6,75-12,6)×109/L versus 7,12 (4,50-9,08)×109/L (U = 96,5; p < 0,05); significantly higher mean group level of C-reactive protein: 120.09 (45.08-164.38) mg/L versus 54.89 (31.14-96.86) mg/L (U = 101.0; p < 0.05); group mean fibrinogen and D-dimer were significantly higher: 7.03 (5.89-8.28) g/L versus 5.98 (4.25-6.80) g/L (U = 99.0; p < 0.05) and 2058.5 (826.0-4026.0) ng/mL versus 982.5 (656.5-1936.0) ng/mL (U = 141.5; p < 0.05) respectively. A higher proportion of individuals with increased prothrombin time was identified: 75.5% (n = 28) versus 32.9% (n = 26) (χ2 = 6.31; p < 0.05). In patients with COVID-19 and PE there was a direct moderate relationship between CRP and D-dimer values (ρ = 0.66; p < 0.05), a direct moderate relationship between CRP and fibrinogen values (ρ = 0.61; p < 0.05). Conclusion. Patients with new coronavirus infection COVID-19 and PE had a marked increase of inflammatory and thrombotic markers (leukocyte count, CRP, fibrinogen, D-dimer). The relationship between the values of CRP, fibrinogen, D-dimer indicates the association of inflammation with the thrombosis markers level.

Complement C5a and Clinical Markers as Predictors of COVID-19 Disease Severity and Mortality in a Multi-Ethnic Population.

Coronavirus disease-2019 (COVID-19) was declared as a pandemic by WHO in March 2020. SARS-CoV-2 causes a wide range of illness from asymptomatic to life-threatening. There is an essential need to identify biomarkers to predict disease severity and mortality during the earlier stages of the disease, aiding treatment and allocation of resources to improve survival. The aim of this study was to identify at the time of SARS-COV-2 infection patients at high risk of developing severe disease associated with low survival using blood parameters, including inflammation and coagulation mediators, vital signs, and pre-existing comorbidities. This cohort included 89 multi-ethnic COVID-19 patients recruited between July 14th and October 20th 2020 in Doha, Qatar. According to clinical severity, patients were grouped into severe (n=33), mild (n=33) and asymptomatic (n=23). Common routine tests such as complete blood count (CBC), glucose, electrolytes, liver and kidney function parameters and markers of inflammation, thrombosis and endothelial dysfunction including complement component split product C5a, Interleukin-6, ferritin and C-reactive protein were measured at the time COVID-19 infection was confirmed. Correlation tests suggest that C5a is a predictive marker of disease severity and mortality, in addition to 40 biological and physiological parameters that were found statistically significant between survivors and non-survivors. Survival analysis showed that high C5a levels, hypoalbuminemia, lymphopenia, elevated procalcitonin, neutrophilic leukocytosis, acute anemia along with increased acute kidney and hepatocellular injury markers were associated with a higher risk of death in COVID-19 patients. Altogether, we created a prognostic classification model, the CAL model (C5a, Albumin, and Lymphocyte count) to predict severity with significant accuracy. Stratification of patients using the CAL model could help in the identification of patients likely to develop severe symptoms in advance so that treatments can be targeted accordingly.

10 Pharmacodynamic profiles of aspirin versus dual-pathway inhibition with either aspirin or clopidogrel among patients with stable atherosclerotic disease

Abstract Aims Inhibition of thrombin-mediated signalling processes using a vascular dose of rivaroxaban in adjunct to single antiplatelet therapy, known as dual-pathway inhibition (DPI), reduces atherothrombotic events in patients with stable atherosclerotic disease. However, there is limited data on the pharmacodynamic (PD) effects of this strategy. Methods This investigation was conducted in selected cohorts of patients (n = 40) with stable atherosclerotic disease enrolled from a larger prospective, open-label, parallel-group PD study who were treated with either aspirin, aspirin plus rivaroxaban 2.5 mg/bid or clopidogrel plus rivaroxaban 2.5 mg/bid. Multiple PD assays providing a comprehensive assessment of markers of thrombosis were used. The primary endpoint was the comparison between groups of platelet-mediated global thrombogenicity by light transmittance aggregometry (LTA) following stimuli with collagen-related peptide + adenosine diphosphate + tissue factor (CATF). Results There were no differences in the primary endpoint between aspirin vs. aspirin plus rivaroxaban 2.5 mg/bid (P = 0.110), aspirin vs. clopidogrel plus rivaroxaban 2.5 mg/bid (P = 0.611) or aspirin plus rivaroxaban 2.5 mg/bid vs. clopidogrel plus rivaroxaban 2.5 mg/bid (P = 0.315). Rivaroxaban-based treatments significantly reduced markers of thrombin generation (peak thrombin and thrombin velocity index). Clopidogrel-based treatments reduced markers of P2Y12 signalling (LTA ADP 20 max and VerifyNow). Aspirin-based treatments reduced markers of markers of cyclooxygenase-1 activity (LTA collagen). Conclusions Compared with aspirin alone, DPI with either aspirin or clopidogrel might provide superior ischaemic protection by targeting pathways alternative to those affected by antiplatelet agents with only a moderate trade-off in bleeding as supported by similar platelet-mediated global thrombogenicity between treatments.

302. Using Antiphospholipid Antibody Presence as an Additional Biomarker to Identify COVID-19 Positive Patients with High Risk for Thrombosis

BackgroundPatients who are hospitalized with Coronavirus 2019 (COVID-19) are known to have increased risk for thrombosis. Several mechanisms have been proposed for increased thrombogenesis, including antiphospholipid antibodies (APLs). We sought to better understand the relationship between a commonly used marker of thrombosis, D-dimer, and antiphospholipid antibodies in relation to thrombosis in COVID-19. MethodsThis was a single-center prospective cohort study. Participants were adults admitted to the hospital with COVID-19 between March and December of 2020. Included patients required a positive COVID-19 nasopharyngeal nucleic acid amplification testing (NAAT), coagulation studies, and regular assessment of D-dimer levels. Patients who were excluded were pregnant adults, use of oral anticoagulants prior to admission, and absence of a positive COVID-19 nasopharyngeal NAAT. We tested 52 patients for antiphospholipid antibodies (APLs), including lupus anticoagulant (LA), anti-beta-2 glycoprotein antibodies (B2GP), and anti-cardiolipin antibodies (aCL). The endpoint for analysis was hospital discharge or development of a confirmed thrombosis. ResultsTwenty-nine of fifty-two patients (55.7%) with COVID-19 had non-negative APLs. Of these patients, twenty-seven (93.1%) had non-negative aCLs, the majority of which were IgM antibodies. There was a total of 7 thrombotic events in our cohort. The sensitivity of D-dimer alone was 85% and the sensitivity of APLs alone was 71%. In patients with an intermediate D-dimer level (i.e., greater than 2 milligrams per liter (mg/L) but less than 5 mg/L), the addition of non-negative APLs increased the sensitivity of D-dimer to 100%. In patients with a high D-dimer (i.e., greater than 5), the combined sensitivity of D-dimer and APLs was 60%. Out of the 7 thrombotic events in our cohort, two patients had negative APLs, however both patients had a D-dimer of greater than 5 mg/L. ConclusionThe use of APLs can assist in risk-stratifying patients in an intermediate-risk D-dimer group to consider prophylactic anticoagulation if APLs are negative and to consider therapeutic anticoagulation if APLs are non-negative. In the high-risk group (i.e., a D-dimer greater than 5 mg/dL), a therapeutic anticoagulation approach may be more appropriate. Disclosures All Authors: No reported disclosures

NEUT-SFL in Patients with COVID-ARDS: A Novel Biomarker for Thrombotic Events?

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an enveloped RNA virus first identified in December 2019 in Wuhan, China, and responsible for coronavirus disease 2019 (COVID-19). The ongoing COVID-19 pandemic is impacting healthcare worldwide. Patients who develop coagulopathy have worse outcomes. The pathophysiology of COVID-19 suggests a strong interplay between hemostasis and immune cells, especially neutrophils. Our purpose was to assess neutrophil fluorescence as a potential biomarker of deep vein thrombosis (DVT) in patients with COVID-acute respiratory distress syndrome (COVID-ARDS). Sixty-one patients with COVID-ARDS admitted to the four intensive care units (ICUs) of a French general hospital were included in this prospective study. Neutrophil activation was assessed by measuring neutrophil fluorescence (NEUT-Side Fluorescence Light, NEUT-SFL) with a specific fluorescent dye staining analyzed by a routine automated flow cytometer Sysmex XN-3000™ (Sysmex, Kobe, Japan). DVT was diagnosed by complete duplex ultrasound (CDU). We found that NEUT-SFL was elevated on admission in patients with COVID-ARDS (49.76 AU, reference value 46.40 AU, p < 0.001), but did not differ between patients with DVT (49.99 AU) and those without (49.52 AU, p = 0.555). NEUT-SFL is elevated in patients with COVID-ARDS, reflecting neutrophil activation, but cannot be used as a marker of thrombosis. Because neutrophils are at interface between immune response and hemostasis through release of neutrophil extracellular traps, monitoring their activation could be an interesting approach to improve our management of coagulopathy during COVID-ARDS. Further research is needed to better understand the pathophysiology of COVID-19 and identify high-performance biomarkers.

Abstract 12228: Metformin Use in Patients Hospitalized With COVID-19: Lower Inflammation, Oxidative Stress, and Thrombotic Risk Markers and Better Clinical Outcomes

Introduction: Diabetes mellitus (DM) is associated with a greater risk of COVID-19 and an increased mortality when the disease is contracted. Metformin use in patients with DM is associated with less COVID-19-related mortality, but the underlying mechanism behind this association remains unclear. Hypothesis: We hypothesize that inhibitory effect of metformin on markers of inflammation, oxidative stress, and hypercoagulability is associate with better clinical outcomes. Methods: Patients with DM on metformin (n=34) and metformin naïve (n=41), and patients without DM (n=73) were enrolled within 48 hours of hospital admission and were diagnosed with COVID-19 by reverse transcription-polymerase chain reaction assay. Laboratory assessments were conducted within 48 hours of hospital admission. The chi-squared test was used to determine whether there was a significant difference in frequencies between disease groups; p&lt; 0.05 was considered a significant difference between groups. Results: DM patients on metformin compared to naïve patients had a lower white blood cell count (p=0.02), d-dimer (p=0.04), urinary 11-dehydro thromboxane B 2 (p=0.01) and urinary liver-type fatty acid binding protein (p=0.03) levels and had lower sequential organ failure assessment score (p=0.0002), intubation rate (p=0.03), and a trend for fewer hospitalized days (p=0.13), lower in-hospital mortality (p= 0.12) and lower mortality plus nonfatal thrombotic event occurrences (p=0.10). Patients on metformin had similar clinical outcomes compared to patients without DM. Conclusion: In conclusion, metformin treatment in COVID-19 patients with DM was associated with lower markers of inflammation, renal ischemia, and thrombosis; and fewer hospitalized days and intubation requirement. Further focused studies are required to support these findings.

Abstract 12602: Local NET Burden is Associated With Biomarkers of Thrombosis and Cardiac Damage in Left Atrial Cardiopathy

Atrial cardiopathy results in disturbed blood flow, providing an ideal thrombogenic environment. Neutrophils promote large vessel occlusions and microthrombosis via release of neutrophil extracellular traps (NETs) and thus lie at the interface of (sterile) inflammation, thrombosis, and fibrosis. We profiled NETs, cytokines, thrombotic factors, and cardiac damage markers in left atrial (LA) cardiopathies together with LA structural analysis. A total of 71 patients (median age 66, 46.5% male) undergoing catheterization procedures with atrial transseptal access at the Freiburg Heart Center were enrolled in this prospective study. Paired samples were collected from peripheral blood draws or from within the left atrium (LA). NET biomarkers, sP-selectin, PAD4, proinflammatory cytokines, thrombotic factors (D-dimers, VWF, ADAMTS13), and cardiac damage markers (TropT, proBNP, ICTP) were measured in plasma or serum as appropriate. Echocardiography was performed to assess LA structure and left ventricular function. Data were analyzed by the type of procedure (MitraClip (MC), left atrial appendage closure (LAAC), pulmonary vein ablation (PVA), or patent foramen ovale closure (PFOC), and by atrial fibrillation (Afib) status at time of procedure. NETs, but not other markers, were elevated in local LA samples as compared to peripheral blood. Conversely, most thrombotic, inflammatory, and cardiac damage markers, but not NETs, were elevated in MC or LAAC as compared to PFOC or PVA patients. Across all patients, NET biomarkers positively correlated with IL-6/TNF-α, VWF, and cardiac damage biomarkers. PAD4/NETs negatively correlated with ADAMTS13 activity, indicating that citrullination may contribute to the prothrombotic phenotype of these patients. Afib was not key to these observations. NETs are locally elevated within the left atrium, with elevation of thrombosis markers including VWF in patients with a diseased left atrium. These elevations partly correlate with structural changes of the left atrium and, therefore, may point toward a disturbed local blood flow. This may provide insight on mechanisms that drive detrimental left atrial remodeling as well as clues regarding thromboembolic risks that are rooted in LA changes rather than Afib.

COVID-19: OBSERVATIONS ON STANDARD TREATMENT ALGORITHMS

In this article, a retrospective analysis of the medical histories was carried out in order to develop an individualized approach to treating COVID-19. Questions were raised about the advisability of the universal immunosuppressive therapy, the need to prescribe glucocorticosteroids only taking into account the cytokine profile of patients, the relationship of glucocorticosteroid therapy and carbohydrate metabolism disorders, as one of the most common complications of pharmacotherapy COVID-19 in the patients we studied. Unjustified immunosuppression in the stage of active fight against the infectious agent as a factor of chronization of the process and decrease of the body reactivity, as well as neutralization by immunosuppressive therapy of factors of protection of innate non-specific immunity mobilized to fight the causative agent. The importance of taking into account the probability of thrombosis with consideration of coagulogram indices and thrombosis markers (D-dimer, fibrin degradation products) does not exclude the risk of the hemorrhagic complications when prescribing high doses of anticoagulants. Antibacterial therapy for uncomplicated viral infections is neither an etiologic nor a pathogenetic treatment option. In addition, it leads to the emergence of resistant strains of microorganisms as a result of mutational variability and evolutionary adaptations of bacteria, which greatly complicates the choice of an effective antibacterial drug in some cases. Antithrombotic, glucocorticosteroid and antibacterial therapy should be clinically and laboratory justified in each individual case. The article focuses on the safety of prescribing antipodagric agents in the treatment of COVID-19 as anti-inflammatory. The review considers all the above-mentioned aspects of COVID-19 therapy with reference to foreign studies, describes options for the pathophysiological development of infection. We note the need to develop systematic data on coronavirus, changes that the pathogen induces in the body. This will allow the development of innovative and effective therapeutic strategies for the treatment of the new coronavirus infection.

Stroke mechanisms associated with coronavirus disease (COVID-19)

The purpose is to identify the pathophysiological mechanisms of stroke development in patients with COVID-19.MethodsA total of 92 patients (46.72+1.72 years) with impairments of cerebral circulation due to COVID-19 (confirmed by positive PCR test results) had been examined. Among them, 67 patients had an ischemic stroke, 14–a transient ischaemic attack, 11–an intracerebral hemorrhage, 3– a subarachnoid hemorrhage.The parameters of hemostasis were measured by standard methods, electrical and viscoelastic parameters of red blood cells (RBC) by the dielectrophoresis method was carried out.ResultsMost of the surveyed persons (60 patients) showed signs of an intravascular coagulation and thrombosis: accelerated platelet-leukocyte aggregation, increased levels of coagulation products, reduced fibrinolysis activity (p=0.002–0.04). The levels of D-dimer, fibrinogen, ESR, platelet count were significantly higher in this group compared to patients in the second one (p<0.01). A moderate increase in summarized rigidity, viscosity of RBC was noted. The level of RBC hemolysis was associated with platelet count (r=0.727, p<0.05), D-dimer level (r=0.422, p=0.04), and fibrinogen level (r=0.318, p<0.05).In the second group of patients (32 persons with a predominance of men - 25 male subjects), the markers of thrombosis had moderate deviations. A sharply reduced RBC deformability with increased summarized rigidity and viscosity was dominant coupled with the background of high electrical conductivity of cell membranes compared to the indicators in the first group (p<0.01). There was a decrease in membrane capacity, surface charge, electrical dipole moment of the cells and polarizability at all frequencies of the electric field than those in the first group (p=0.0001–0.05). A sharp decrease in RBC deformability creates obstacles to overcoming small-diameter capillaries, leading to violations of microcirculatory blood flow.The RBC deformability was associated with levels of ferritin (r=0.407, p=0.024), HbA1c (r=0.419, p=0.033), uric acid (r=−0.303, p<0.05), and LDL cholesterol (r=0.426, p=0.029).Incubation of blood samples in vitro for 10 min with riboflavin, nicotinamide, inosine, which ensures RBC energy metabolism (anaerobic glycolysis and pentose phosphate pathway) restored the reduced the RBC deformability (p<0.01), and decreased the RBC aggregation (p<0.001). Similar changes were obtained when coenzymes were administered to patients in vivo.ConclusionTwo independent mechanisms of cerebral circulatory disturbances have been identified in COVID-19 stroke patients: both thrombotic and hemorheologic. The thrombotic variant is associated with a procoagulant state and with an activity of inflammation. The hemorheologic variant is caused by a decrease in the activity of the RBC energy metabolism enzymes; it is associated with the presence of metabolic disorders and is to be eliminated by an administration of coenzymes and metabolites.FUNDunding AcknowledgementType of funding sources: Other. Main funding source(s): This research was done within the framework of the topic “Epidemiological monitoring of the population state of health and studies on molecular genetic and molecular biological mechanisms of the development of common internal diseases in Siberia for improvement of the relevant diagnostic, preventive, and therapeutic methods” in State Assignment No. 0324-2018-0001, registration No. AAAA-A17-117112850280-2

Abstract P260: SARS-Cov-2 Spike Protein S1-Mediated Endothelial Injury And Pro-Inflammatory State Is Amplified By Dihydrotestosterone And Prevented By Mineralocorticoid Antagonism

Objectives: Men are disproportionately affected by the coronavirus disease-2019 (COVID-19) and experience higher mortality as compared to women. Endothelial dysfunction has been proposed as a major inciting factor of pro-inflammatory and thrombotic changes in COVID-19 infection. The vascular effects of androgen signaling and inflammatory cytokines in severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)-mediated endothelial damage are not defined. Methods: Here, we determined the effects of SARS-CoV-2 spike protein-mediated endothelial damage under conditions of exposure to androgens and TNF-α. We tested the therapeutic effects of approved drugs: angiotensin receptor blockade by valsartan and mineralocorticoid receptor antagonism by spironolactone. Circulating endothelial injury markers VCAM-1 and E-selectin were measured in the plasma from men and women diagnosed with COVID-19. Results: Exposure of endothelial cells (ECs) in vitro to androgen dihydrotestosterone (DHT) exacerbated spike protein S1-mediated endothelial injury transcripts for cell adhesion molecules E-selectin, VCAM-1, and ICAM-1 and anti-fibrinolytic PAI-1 (P&lt;0.05), as well as increased THP-1 monocyte adhesion to ECs (P=0.032). Spironolactone dramatically reduced DHT+S1-induced endothelial activation. Pre-treatment of ECs with valsartan did not increase endothelial activation by S1. TNF-α exacerbated S1-induced EC activation and spironolactone abrogated this effect. Analysis of plasma from patients hospitalized with COVID-19 showed concordant higher VCAM-1 in men as compared to women. Conclusions: Androgen exposure promoted spike protein-mediated endothelial injury by increasing markers of inflammation and thrombosis. A beneficial therapeutic effect of the FDA-approved drug spironolactone was observed, supporting a rationale for further evaluation as an adjunct treatment in COVID-19.

The Angiotensin Converting Enzyme Deletion/Deletion Genotype Is a Risk Factor for Severe COVID-19: Implication and Utility for Patients Admitted to Emergency Department.

Background and objective: Insertion/deletion polymorphisms of angiotensin-converting enzyme (ACE) have been previously described in association with adult respiratory distress syndrome (ARDS) and correlated to outcome. The ACE deletion/deletion(D/D)genotype represents a marker of thrombosis in subjects apparently without predisposing factors and/or traditional thrombophilic alterations and increases the risk of venous thromboembolism in subjects in whom a thrombogenic condition occurs. Thrombosis seems to play a role very early in the disease caused by SARS-CoV-2, in particular in those with severe COVID-19 pneumonia. The counterbalance between angiotensin-converting enzyme (ACE) and ACE2 activities in COVID-19 disease may play a crucial role in the thrombo-inflammatory process. We hypothesised that a genetic predisposition could condition the severity and complications of SARS-CoV-2 infection. Materials and methods: We conducted a spontaneous, single centre observational study in the Sub-Intensive Care Unit of A.O.R.N. Ospedali dei Colli, Cotugno Hospital, Naples (Italy). In this study, we performed genetic screening for ACE D/D genotype and other thrombophilic mutations in 20 patients affected by ARDS related to COVID-19 pneumonia, compared to 19 age- and sex-matched healthy controls. Results: All tested patients had multiple polymorphisms and, in particular, a significantly higher prevalence of ACE D/D polymorphism in severe COVID-19 patients Conclusion: We found that the majority of patients who tested positive for ACE D-D genotype and who were not associated with other risk factors for VTE showed an evolution to ARDS. This finding could have a predicting role in the selection of patients more prone to developing severe COVID-19 during clinical observation in emergency department.

Obesity, inflammatory and thrombotic markers, and major clinical outcomes in critically ill patients with COVID-19 in the US.

This study aimed to determine whether obesity is independently associated with major adverse clinical outcomes and inflammatory and thrombotic markers in critically ill patients with COVID-19. The primary outcome was in-hospital mortality in adults with COVID-19 admitted to intensive care units across the US. Secondary outcomes were acute respiratory distress syndrome (ARDS), acute kidney injury requiring renal replacement therapy (AKI-RRT), thrombotic events, and seven blood markers of inflammation and thrombosis. Unadjusted and multivariable-adjusted models were used. Among the 4,908 study patients, mean (SD) age was 60.9 (14.7) years, 3,095 (62.8%) were male, and 2,552 (52.0%) had obesity. In multivariable models, BMI was not associated with mortality. Higher BMI beginning at 25 kg/m2 was associated with a greater risk of ARDS and AKI-RRT but not thrombosis. There was no clinically significant association between BMI and inflammatory or thrombotic markers. In critically ill patients with COVID-19, higher BMI was not associated with death or thrombotic events but was associated with a greater risk of ARDS and AKI-RRT. The lack of an association between BMI and circulating biomarkers calls into question the paradigm that obesity contributes to poor outcomes in critically ill patients with COVID-19 by upregulating systemic inflammatory and prothrombotic pathways.

DIAGNOSTIC VALUE OF INTERLEUKIN-6 AND D-DIMER IN INFLAMMATORY BOWEL DISEASES

The article deals with the diagnostic value of changes in the content of interleukin-6 and D-dimer in patients with inflammatory bowel diseases (IBD) - Ulcerative Colitis (UC) and Crohn Disease (CD).&#x0D; Materials and methods: We have studied 34 patients with UC and 18 people with CD with continuously recurrent course. There are also analysed data of 15 patients with UC and 15 patients with CD in period of the remission. The control group included 30 healthy volunteers. Clinical, endoscopic examination, an immunoassay determination of the content of interleukin-6 (IL-6) and D-dimer, was carried out. CD activity was assessed using the Chron’s disease activity index. The localization of CD was established according to the Montreal classification. The clinical activity of the disease in patients with UC was determined using the classification according to Truelove-Witts. The value of the index consisted of the sum of points for each indicator, which allowed determining the activity of the inflammatory process.&#x0D; Statistical processing was performed using STATISTICA 10.0 (StatSoft. Inc., USA), at p&lt;0.05, the discrepancies between the obtained data were considered statistically significant.&#x0D; Research results. According to the obtained data, mild activity was found in 7 patients with UC (20.6%) and 4 - with CD (22.3%), moderate severity - in 17 (50.0%) and 8 (44.4%) patients, severe - in 10 (29.4%) and 6 (33.3%) patients. In the remission stage, 15 patients with UC and 15 patients with CD were examined. According to the Montreal classification, lesions of the upper gastrointestinal tract (L4) were observed in 4 patients with CD (22.2%), terminal ileitis (L1) occurred in 10 patients (55.6%), 4 patients were diagnosed with ileocolitis (L3) - 22.2%.&#x0D; It has been established that patients with an active course of UC and CD had a higher level of proinflammatory markers and thrombosis markers (D-dimer and IL-6) compared with inactive course and control group (p&lt;0.05). The D-dimer content in UC was 690±315 pg/l. It is confirmed the increase of the content of this marker in 2.55 times compared to control group and in 1.43 times than inactive course of the disease respectively. The level of IL-6 at active in course of UC was 3.36±1.78 pg/ml, increasing in 2.07 times against control and in 1.72 times compared with group of remission.&#x0D; In the active course of CD, the level of D-dimer reached 720±267 pg/ml, increasing in 2.67 times in relation to control and in 1.60 times with respect to patients under remission. The content of IL-6 in the active course of CD reached 4.07±2.17 pg/ml, increasing in 2.85 times in comparison to healthy individuals and in 2.83 times relatively patients with remission.&#x0D; While using the ROC-analysis, the most sensitive value as a “cut point” for the diagnosis of the active process in the course of IBD can be considered the content of the D-dimer in an amount of more than 650 pg/l. The calculated AUC in ROC-analysis for the IL-6 content was 73.7%±6.14% (61.7 - 85.7, p&lt;0.001), indicating the prognostic value of the model. The values of the IL-6 content of more than 2.80 pg/ml are diagnostically significant to predict the active process in IBD.

Biomarkers of Platelet Activation and Their Prognostic Value in Patients in Sepsis Associated Coagulopathy

Introduction:Sepsis-associated disseminated intravascular coagulation (SAC) is associated with marked hemostatic changes including transient thrombocytopenia due to their enodogenous activation / consumption. Platelets are decreased in SAC and exhibit compromised function. Patients with SAC are at an increased risk of thrombosis and / or bleeding. The widespread activation of platelets contribute to vascular occlusions, fibrin deposition, multi-organ dysfunction, contributing to a two-fold increase in mortality. At this time there is no single biomarker or laboratory test that can effectively diagnose SAC; however, the International Society on Thrombosis and Hemostasis (ISTH) has introduced a scoring system based on laboratory parameters such as platelet count, clotting time, and levels of markers of thrombosis to calculate SAC severity in terms of DIC score. Measurement of activation markers that describe the interplay between inflammation and platelet function may provide insight into the pathogenesis of SAC. The purpose of this study was to measure markers of platelet function in the plasma of patients with clinically established SAC and to determine their association to disease severity and outcome.Materials and Methods:Plasma samples from 103 adult intensive care unit (ICU) patients with sepsis and suspected disseminated intravascular coagulation (DIC) were acquired from the University of Utah Hospital at the time of ICU admission as well as on days 4 and 8. Samples with relevant clinical information were transported to Loyola University Chicago under an IRB-approved protocol and stored at -80˚C prior to analysis. Plasma samples from healthy individuals (n=50) were purchased from George King Biomedical (Overland, KS). DIC scores were calculated using the ISTH scoring algorithm which incorporates platelet count, D-Dimer, INR, and fibrinogen. Patients were categorized as having no DIC, non-overt DIC, or overt DIC. Plasma levels of CD40L, von Willebrand Factor (vWF), platelet factor-4 (PF-4), and microparticles (MP) were quantified using commercially available ELISAs performed according to manufacturer instructions.Results:Markers of platelet activation were significantly elevated in patients with sepsis alone and with suspected DIC compared to normal healthy individuals on ICU day 0 (p<0.001). Levels of platelet-associated biomarkers were compared between survivors and non-survivors. PF-4 was significantly decreased in non-survivors compared to survivors (p = 0.0156). No other significant variations based on survival or based on DIC score category were observed.Since platelet count is often dramatically reduced in patients with DIC, the relationship between platelet count and levels of markers of platelet activity was also evaluated. Patients were stratified based on platelet count and levels of markers were compared between groups. CD40L, vWF, PF4, and MP showed significant variation based on platelet count, with all markers exhibiting stepwise elevation with increasing platelet count (Table 1).Conclusion: Markers of platelet activation were significantly elevated in patients with SAC compared to healthy individuals. This increase was independent of platelet count, underscoring the importance of activation processes to this disease process. Of the markers studied, PF4 showed significant difference based on DIC score or mortality, and differentiated the non-survivors compared to survivors. Levels of CD40L, vWF, PF4, and MP showed significant association with platelet count, increasing in a stepwise manner with increases in platelet count. These studies also suggest that the markers of platelet activation in SAC are primarily regulated by the number of circulating platelets and may be independent of the factors leading to their endogenous consumption. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Modulated molecular markers of restenosis and thrombosis by in-vitro vascular cells exposed to bioresorbable scaffolds

Drug-eluting bioresorbable vascular scaffolds (BVSs) have emerged as a potential breakthrough for the treatment of coronary artery stenosis, providing mechanical support and drug delivery followed by complete resorption. Restenosis and thrombosis remain the primary limitations in clinical use. The study aimed to identify potential markers of restenosis and thrombosis analyzing the vascular wall cell transcriptomic profile modulation triggered by BVS at different values of shear stress (SS). Human coronary artery endothelial cells and smooth muscle cells were cultured under SS (1 and 20 dyne cm−2) for 6 h without and with application of BVS and everolimus 600 nM. Cell RNA-Seq and bioinformatics analysis identified modulated genes by direct comparison of SS conditions and Gene Ontology (GO). The results of different experimental conditions and GO analysis highlighted the modulation of specific genes as semaphorin 3E, mesenchyme homeobox 2, bone morphogenetic protein 4, (heme oxygenase 1) and selectin E, with different roles in pathological evolution of disease. Transcriptomic analysis of dynamic vascular cell cultures identifies candidate genes related to pro-restenotic and pro-thrombotic mechanisms in an in-vitro setting of BVS, which are not adequately contrasted by everolimus addition.

Beneficial Effects of Mineralocorticoid Receptor Pathway Blockade against Endothelial Inflammation Induced by SARS-CoV-2 Spike Protein.

Background: Vascular endothelial cells activation and dysfunction mediate inflammation and abnormal coagulation in COVID-19 patients. Mineralocorticoid receptor (MR) signaling and its downstream target Galectin-3 (Gal-3) are known to mediate cardiovascular inflammation and might be involved in the pathogenesis of COVID-19 complications. Accordingly, we aimed to investigate the potential beneficial effects of MR antagonism and Gal-3 inhibition on the inflammatory response induced by SARS-CoV-2 Spike protein in human aortic endothelial cells (HAECs). Methods: HAECs were treated with recombinant SARS-COV2 Spike (S) protein. MR antagonists (namely spironolactone and eplerenone) or the Gal-3 inhibitor G3P-01 were supplemented before and after S protein challenge. HAECs supernatants were assessed by ELISA or Western blotting. Results: HAECs treated with recombinant S protein resulted in enhanced secretion of inflammatory molecules (interleukin-6, monocyte chemoattractant protein-1, interleukin-18, interleukin-27, and interferon-γ) as well as in the thrombosis marker plasminogen activator inhibitor (PAI)-1. This was prevented and reversed by both MR antagonists and G3P-01. Conclusions: These findings indicate that MR/Gal-3 pathway blockade could be a promising option to reduce endothelial inflammation in SARS-CoV-2 infection.

Stroke Mechanism in COVID-19 Infection: A Prospective Case-Control Study

BackgroundThe characteristics and pathophysiological mechanisms involved in acute ischemic stroke in patients with COVID-19 infection have not been fully clarified. We prospectively studied the phenotypic and etiological features of acute stroke occurring in COVID-19 infection. Patients & methodsWithin nine months starting from April-2020, the presence of COVID-19 infection was determined by thoracic CT and SARS-CoV-2 PCR in all acute stroke cases managed in a single tertiary center. Consecutive and prospective data on vascular risk factors/comorbidities, in-hospital quality metrics, discharge outcomes, etiological subclassification and blood markers of thrombosis / inflammation were compared in 44 COVID-19 positive cases (37 acute ischemic stroke, 5 TIA, 2 intracerebral hematoma) and 509 COVID-19 negative patients (355 ischemic, 105 TIA, 44 hematoma and 5 stroke mimic). ResultsCOVID-19 positive patients had more severe strokes, delayed hospital admission, longer hospital stay, higher mortality rates, but had similar vascular risk factors/comorbidities frequency, thrombolysis/thrombectomy utilization rates, metrics, and stroke etiological subtype. They had significantly higher CRP, fibrinogen, ferritin, leukocyte count and lower lymphocyte count. No difference was detected in aPTT, INR, D-dimer, platelet, hemoglobin, homocysteine levels and ANA, anti-dsDNA antibody and ENA panel positivity rates. Anti-phospholipid antibodies have been studied in 70% of COVID-19 positive and all cryptogenic patients, but were never found positive. Tests for coagulation factor levels and hereditary thrombophilia did not show major thrombophilia in any of the stroke patients with COVID-19. ConclusionWe documented that there is no significant difference in etiological spectrum in acute stroke patients with COVID-19 infection. In addition, cryptogenic stroke and antiphospholipid antibody positivity rates did not increase.

High Levels of Neutrophil Extracellular Traps Persist in the Lower Respiratory Tract of Critically Ill Patients With Coronavirus Disease 2019.

Lower respiratory tract (LRT) disease induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can deteriorate to acute respiratory distress syndrome (ARDS). Because the release of neutrophil extracellular traps (NETs) is implicated in ARDS pathogenesis, we investigated the presence of NETs and correlates of pathogenesis in blood and LRT samples of critically ill patients with COVID-19. Plasma NET levels peaked early after intensive care unit admission and were correlated with the SARS-CoV-2 RNA load in sputum and levels of neutrophil-recruiting chemokines and inflammatory markers in plasma samples. The baseline plasma NET quantity was correlated with disease severity but was not associated with soluble markers of thrombosis or with development of thrombosis. High NET levels were present in LRT samples and persisted during the course of COVID-19, consistent with the detection of NETs in bronchi and alveolar spaces in lung tissue from deceased patient with COVID-19. Thus, NETs are produced and retained in the LRT of critically ill patients with COVID-19 and could contribute to SARS-CoV-2-induced ARDS disease.

Uric Acid-An Emergent Risk Marker for Thrombosis?

Hyperuricemia is nowadays an established cardiovascular risk factor. Experimental studies linked elevated serum uric acid (SUA) levels with endothelial dysfunction (ED), inflammation, and prothrombotic state. The purpose of this review is to summarize the current evidence that emphasizes the possible role of uric acid as a biomarker for a prothrombotic state. A large number of clinical trials correlated SUA levels with both incident and recurrent cases of venous thromboembolism (VTE), independent of other confounding risk factors. Moreover, increased SUA levels may be an important tool for the risk stratification of patients with pulmonary embolism (PE). Left atrial thrombosis was correlated with high SUA levels in several studies and its addition to classical risk scores improved their predictive abilities. In patients with acute myocardial infarction (MI), hyperuricemia was associated with increased mortality, and the idea that hyperuricemia may be able to act as a surrogate to unstable coronary plaques was advanced. Finally, SUA was correlated with an increased risk of thromboembolic events in different systemic diseases. In conclusion, uric acid has been considered a marker of a thrombotic milieu in several clinical scenarios. However, this causality is still controversial, and more experimental and clinical data is needed.

Anticardiolipin and other antiphospholipid antibodies in critically ill COVID-19 positive and negative patients

BackgroundReports of severe COVID-19 being associated with thrombosis, antiphospholipid antibodies (APLA), and antiphospholipid syndrome have yielded disparate conclusions. Studies comparing patients with COVID-19 with contemporaneous controls of similar severity are...