Altered Coagulation Research Articles

Altered platelet reactivity, coagulation, endothelial and inflammatory markers early after smoking cessation verified with cotinine plasma concentration

Abstract Background/Introduction Cigarette smoking is a potent modifiable risk factor for coronary artery disease (CAD) including patients after percutaneous coronary interventions (PCI). Cigarette smoking is associated with major changes in platelet, endothelial, inflammatory, and coagulation responses. The early period after PCI witnesses the highest risk of thrombotic events. However, data regarding alterations of prothrombotic state and platelet reactivity early after smoking cessation are scarce. Purpose We investigated alterations to platelet reactivity, coagulation and markers of platelet, endothelial and neutrophil activation in clopidogrel-treated patients with CAD after PCI before and after smoking cessation. Methods Patients aged 18 years or older, who smoked at least 10 cigarettes per day, at least 30 days after successful PCI with stent placement, and those treated with 75 mg clopidogrel and 75 mg acetylsalicylic acid (ASA) once daily, were included in the study and were encouraged to quit the habit. Platelet reactivity was measured with VerifyNow system and the concentrations of cotinine, thrombomodulin, P-selectin, platelet factor 4 (CXCL4/PF4), citrullinated histone H3 (H3cit), were assessed in blood samples at baseline and at the 30-day follow-up visit. Smoking cessation was defined as cotinine concentrations of <50 ng/ml. Results Among 117 patients, being current smokers, who attended the baseline visit, 84 patients completed a 30-day follow-up. The median [interquartile range] age of patients was 50 [55–65] years, median body mass index was 27.55 [25.2–30.9] kg/m2, and a median load of smoking was 40 [30–47] pack-years. At the follow-up visit, 30 patients stopped smoking as confirmed by cotinine concentrations (study group) and 54 patients continued smoking (control group). Baseline characteristics were similar in both groups. In the study group as compared to the control group, at day 30 a change in platelet reactivity was larger (Δ platelet reactivity units (PRU) 19 [2, 43] vs −6 [−32, 37], p=0.018), along with a change in P-selectin concentration (−11.82 [−23.62, 1.34] vs 7.19 [−14.24, 17.19] ng/ml), p=0.005). The change in the concentrations of thrombomodulin, CXCL4/PF4, and citH3 did not differ significantly between groups. Conclusion In the early period after smoking cessation in coronary artery disease patients following PCI treated with clopidogrel and ASA, an increase in platelet reactivity with a decrease in P-selectin levels was observed. No significant changes in other markers were noticed. It might be speculated that the risk of thrombotic complications might be paradoxically enhanced among such patients after smoking cessation despite dual antiplatelet therapy. Funding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): Centre of Postgraduate Medical Education

S1279 Disparities in Outcomes Among Patients With Cirrhosis and Gastrointestinal Bleeding Undergoing Endoscopy

Introduction: Patients with cirrhosis are prone to gastrointestinal (GI) bleeding due to a higher prevalence of varices, and secondary to altered coagulation. The aim of our study is to understand the characteristics of patients with cirrhosis undergoing Endoscopic Treatment (ET) for GI bleeding, and the risk factors associated with mortality. Methods: We explored the 2019 National Inpatient Sample (NIS) from the Healthcare Cost and Utilization Project, Agency for Healthcare Research and Quality, and their partners (https://www.hcup-us.ahrq.gov/nisoverview.jsp) for cases of cirrhosis with a procedural code for “control bleeding in gastrointestinal tract, endo” (classified under endoscopic control of bleeding) (0W3P8ZZ). Patient characteristics were compared using Chi-Square tests and mortality risks were analyzed via multivariate logistic regression. Results: 24,635 patients with cirrhosis undergoing ET for GI bleeding were included in our final analysis. These patients were more likely to be males (59.5%), age ≥ 61 (57.5%), White (67.2%), and covered by Medicare (50.8%). 40.4% were smokers, 33.2% had a history of alcohol abuse and 2% reported marijuana use. 38.8% reported Acute Kidney Injury (AKI), 10.6% had hyperkalemia and 17.2% had hypokalemia. Comorbidities- 37.1% had diabetes, 58.5% had hypertension, 18.0% were obese and 20.5% had chronic pulmonary disease. (Figure) Our study had mortality rate of 9.4%. After adjusting for variables, AKI (77.3%, aOR 5.799, p< 0.01) and hyperkalemia (19.0%, aOR1.516, p< 0.01) showed increased risk of mortality. Racial differences were also noticed as Blacks (aOR 1.142, p=0.027) had a higher death risk as compared to Caucasians. Cardiopulmonary resuscitation was also linked with a poor outcome (aOR 15.821, p< 0.01). Meanwhile, hyperlipidemia (aOR 0.702, p< 0.01), smoking (0.719, p< 0.01), hypokalemia (aOR 0.831, p< 0.01), diabetes (aOR 0.773, p< 0.01), hypertension (aOR 0.694, p< 0.01), obesity (aOR 0.754, p< 0.01), showed a reduced mortality risk. Conclusion: Our study provides a fresh perspective on various risk factors for mortality among patients with cirrhosis undergoing ET for GI bleeding. Physicians should thus be careful and monitor pre and post-procedural occurrences for timely management to improve the outcomes of these patients.Figure 1.: Forest plot showing demographics and risk factors of patients with cirrhosis and GI bleeding.

Science, Medicine, and the Anesthesiologist

Science, Medicine, and the Anesthesiologist

Acquired hemophilia A following COVID-19 vaccination – The importance of prompt diagnosis: A case report

Acquired hemophilia A (AHA) is a rare coagulopathy characterized by hemorrhagic manifestations. It has been linked to various conditions, including autoimmune disorders, drugs, tumors, lymphoproliferative disorders, and infections. We present a case of AHA in a 71-year-old male patient with cutaneous hematoma occurring 8 days after vaccination for COVID-19. This report aims to highlight the risk of FVIII inhibitor development following an immune stimulus, thus improving our knowledge regarding possible vaccination-related adverse events. Furthermore, we underline how the potential risk of not recognizing disease manifestations promptly, together with specific coagulation alterations, could significantly affect the patient's outcome. Adequate management plans and the diffusion of shared guidelines are of fundamental importance in order to prevent the development of life-threatening complications and initiate appropriate treatment as soon as possible. Data AvailabilityAll data generated or analyzed during this study are included in this article. Further inquiries can be directed to the corresponding author.

Hospital Outcomes among COVID-19 Hospitalizations with Acute Ischemic Stroke: Cross-Sectional Study Results from California State Inpatient Database.

Coronavirus disease 2019 (COVID-19) could be a risk factor for acute ischemic stroke (AIS) due to the altered coagulation process and hyperinflammation. This study examined the risk factors, clinical profile, and hospital outcomes of COVID-19 hospitalizations with AIS. This study was a retrospective analysis of data from California State Inpatient Database (SID) during 2019 and 2020. COVID-19 hospitalizations with age ≥ 18 years during 2020 and a historical cohort without COVID-19 from 2019 were included in the analysis. The primary outcomes studied were in-hospital mortality and discharge to destinations other than home. There were 91,420 COVID-19 hospitalizations, of which, 1027 (1.1%) had AIS. The historical control cohort included 58,083 AIS hospitalizations without COVID-19. Conditional logistic regression analysis showed that the odds of in-hospital mortality, discharge to destinations other than home, DVT, pulmonary embolism, septic shock, and mechanical ventilation were significantly higher among COVID-19 hospitalizations with AIS, compared to those without AIS. The odds of in-hospital mortality, DVT, pulmonary embolism, septic shock, mechanical ventilation, and respiratory failure were significantly higher among COVID-19 hospitalizations with AIS, compared to AIS hospitalizations without COVID-19. Although the prevalence of AIS was low among COVID-19 hospitalizations, it was associated with higher mortality and greater rates of discharges to destinations other than home.

An international study on activated partial thromboplastin time prolongation. Part 2: Interpretative commenting

BackgroundProviding evidence-based interpretative comments (IC) is an integral task of clinical laboratory professionals. It may be of special relevance for coagulation testing, where pathological first-line tests could trigger more specialized tests. Our aim was to evaluate the quality of ICs provided to the physician in two samples with activated partial thromboplastin time (APTT) prolongation. Material and methodsTwo lyophilized plasma samples and their respective fictional clinical cases (case 1: heparin contamination and case 2: factor VIII deficiency) were sent to European laboratories for APTT and APTT mixing test measurement, and elaboration of ICs based on their results.The quality of ICs was evaluated in terms of analytical classification, laboratory interpretation, advice to physician, clarity, length and whether the clinical question was answered. ResultsA total of 214 laboratories were included. Classification of the analytical result was stated in 57 % of comments. Laboratory interpretation was found in 91 % of comments for case 1 and 83.3 % for case 2, among which 9.3 % and 6.5 % were considered wrong, respectively. Advice for the requesting physician was provided in 65.8 % of comments for case 1 and 61.2 % for case 2, among which 36 % and 4.7 % were considered wrong, respectively. More than 70 % of comments for both cases were evaluated as clear and of an adequate length. ConclusionA significant number of laboratories provide clear interpretations and helpful advice for the management of altered coagulation results. Nevertheless, the finding of several confusing and misleading comments highlights the need for recommendations on elaboration of interpretative comments.

Correlation of vascular structural changes in a cadaveric model and obesity-related cardiovascular non-communicable diseases

IntroductionCarrying excess body weight is a vital risk factor for obesity-related chronic diseases affecting blood vessels. Obesity influences cardiovascular non-communicable diseases (NCDs) via vascular structural changes, which involve alterations in lipids, blood pressure, coagulation, fibrinolysis, and inflammation, leading to endothelial dysfunction due to vascular remodeling and stiffness. Small peripheral vessels are the first to be impacted; however, it is unclear whether this change is followed by microscopic changes in the aorta. ObjectivesTo determine the correlation of vascular structure with the incidence of NCDs and subcutaneous fat thickness and to study micro-scale changes in vascular structure, especially concerning collagen in the aorta, using a cadaveric model. MethodsTwenty-four cadaveric models were classified into a control group and an NCD group. The subcutaneous fat thickness was measured on the arm, anterior abdomen, and thigh. The aorta was collected and stained with hematoxylin, eosin, and Masson's trichrome for collagen evaluation. The vessel thickness was morphometrically analyzed. Scanning electron microscopy was performed to identify the extracellular matrix organization in the vessel. ResultsDisorganization of the extracellular matrix and fragments of the vascular wall were found in the NCDs group. The tunica intima of the NCDs group represented endothelial dysfunction with macrophage foam cells. The thickness of the tunica intima of the NCDs group slightly increased without being significantly different compared to control group with 144.63 ± 124.38 µm and 105.60 ± 27.49 µm, respectively. However, the thickness of tunica media of the NCDs group significantly decreased compared to control group with 956.58 ± 27.80 µm and 1167.43 ± 48.6 µm, respectively. Collagen deposits in the aortic wall significantly increased by 15% in the NCDs group especially in tunica media by 17.4% compared to control. The results showed a correlation between the amount of collagen fiber and subcutaneous fat on the thigh. ConclusionThere was a change toward irregular microstructural patterns and increased collagen fibers in NCDs. In addition, there was a correlation between collagen fiber density and the subcutaneous fat thickness of the thigh in cadavers with a history of NCDs.

Abstract P2085: Whole Blood Microfluidics To Assess Direct Oral Anticoagulant (doac) Activity And Reversal In Neonatal Cardiac Patients

Introduction: VTEs increase morbidity in neonates with congenital heart disease. Although heparin is the main therapy for prevention / treatment, DOACs are attractive alternatives due to ease of administration, no dependence on antithrombin and wider therapeutic window. Preclinical studies to evaluate their effects on clot formation and antidote reversal are hampered by a lack of low volume assays that permit multiple analyses. Hypothesis: A low volume microfluidic system can reproducibly measure activity and reversibility of DOAC-induced alterations in coagulation. Methods: A microfluidic device was used to study whole blood (WB) coagulation in response to co-immobilized tissue factor and collagen. Blood was drawn before and 24h after surgery. Platelet and fibrin deposition were monitored by fluorescence microscopy (Apixaban ± andexanet). Results: 34 neonates were enrolled. Most participants had double ventricle physiology, required bypass and blood products. There was no difference (P&gt;0.05) in Hb / Hct, platelet count, or fibrinogen levels pre- vs post-surgery. Time to clot-induced channel occlusion and platelet / fibrin deposition were similar in post-operative vs adult participants (7.6±3.1 min vs 7.9±1.5 min; Figure 1A), the former prolonged (14.2±1.9 min) and the latter reduced in pre-operative patients. Although Apixaban reduced clot formation (Figure 1B), it prevented channel occlusion in most post-operative neonates vs adults (60% vs 20%). Andexanet reversed its effects, restoring coagulation to non-treatment levels. Conclusions: WB microfluidics demonstrated that post-operative neonates are hypercoagulable compared to the pre-operative state and are responsive to apixaban, the effects of which can be reversed with Andexanet.

Des bleus mais pas de coups

AbstractA 2-month-old newborn was hospitalized for hemorrhagic picture associated with deterioration of his general condition and eczema. A thrombocytopenia is highlighted, without any other alteration of coagulation. As the patient develops pneumocystis jirovecii infection and signs of vasculitis, immune deficiency is suspected. The discovery of CD8 lymphopenia, microthrombocytopenia, and the presence of eczema, lead to a diagnosis of Wiskott-Aldrich syndrome, then confirmed by molecular biology.

Thrombocytopenia and increased risk of adverse outcome in COVID-19 patients.

BackgroundThrombocytopenia was common in the coronavirus disease 2019 (COVID-19) patients during the infection, while the role of thrombocytopenia in COVID-19 pathogenesis and its relationship with systemic host response remained obscure. The study aimed to systematically evaluate the relationship between thrombocytopenia in COVID-19 patients and clinical, haematological and biochemical markers of the disease as well as adverse outcomes.MethodsTo assess the relationship between abnormal platelet levels and disease progression, a multi-center retrospective cohort study was conducted. COVID-19 patients with thrombocytopenia and a sub-cohort of matched patients without thrombocytopenia were compared for their clinical manifestations, haematological disorders, biochemical parameters, inflammatory markers and clinical outcome.ResultsThrombocytopenia was present in 127 of 2,209 analyzed patients on admission. Compared with the control group, thrombocytopenia patients developed significantly higher frequency of respiratory failure (41.9% vs. 22.6%, P = 0.020), intensive care unit entrance (25.6% vs. 11.5%, P = 0.012), disseminated intravascular coagulation (45.2% vs. 10.6%, P < 0.001), more altered platelet morphology indexes and coagulation perturbation, higher levels of inflammatory markers. In addition, a significantly increased all-cause mortality (hazard ratio 3.08, 95% confidence interval 2.26–4.18, P < 0.001) was also observed in the patients with thrombocytopenia. Late development of thrombocytopenia beyond 14 days post-symptom was observed in 61 patients, from whom a comparable mortality rate yet longer duration to death was observed compared to those with early thrombocytopenia.ConclusionsOur finding from this study adds to previous evidence that thrombocytopenia is associated with adverse outcome of the disease and recommend that platelet count and indices be included alongside other haematological, biochemical and inflammatory markers in COVID-19 patients’ assessment during the hospital stay.

P1675: POST-HOSPITAL DISCHARGE EVALUATION OF COVID-19 SURVIVORS WHO SUFFERED ACUTE VENOUS THROMBOEMBOLISM (VTE) DURING HOSPITALIZATION: THE BERGAMO EXPERIENCE

Background: Acute COVID-19 manifestations are well known, while post-COVID clinical and laboratory consequences are less clear. Due to the strict association of COVID-19 with thrombosis, the Hemostasis Service of Bergamo hospital had a central role in guiding anticoagulant treatments in patients during the acute and post-discharge phases. Aims: In the post-hospital discharge setting, in a cohort of COVID-19 survivors who experienced VTE during the acute phase and were followed up at our outpatient clinic, we aim to evaluate whether alterations in coagulation and inflammatory parameters were occurring and were possibly associated with disease severity. Methods: An outpatient service specifically dedicated to follow-up of COVID-19 survivors was set up at our Institution. A retrospective analysis of a cohort of 1563 patients enrolled from May 2020 to November 2020 was performed. Data were obtained on the occurrence or not of acute COVID-19 -associated VTE, the site of thrombosis, ongoing anticoagulant treatments, and the results of laboratory tests performed at the first post-discharge assessment visit, including blood cell count, D-dimer, fibrinogen, CRP and LDH. The severity degree of COVID-19 at the onset was defined by the need of Continuous Positive Airway Pressure devices (C-PAP) or tracheal intubation as compared to no-oxygen need, or use of nasal flow, reservoir mask, or Venturi Mask. Statistical analyses were performed using SPSS v21 software. Results of lab tests were expressed as median values (5th-95th percentile). Results: Of the 1563 enrolled outpatient COVID-19 survivors [974 M/589 F, median age of 60 years (35.5-81)], 56 subjects [43M/13F; median age 62 years (42-80.5)] had had VTE during COVID-19 acute phase (3.58%), as follows: 9 isolated proximal DVT (4 in upper extremity), 45 isolated PE (34 subsegmental and/or segmental PE; 11 central pulmonary artery thrombosis), 2 DVT+PE. Concerning COVID-19 severity, 33 of 56 patients needed C-PAP or tracheal intubation. At the follow-up visit, patients were still on anticoagulation (76% with DOAC). Laboratory data analyses revealed that 25% of VTE-subjects had increased levels of both D-dimer and fibrinogen, and 11% had elevated levels of CRP, while no abnormalities were observed in the platelet and leucocyte counts. CRP significantly correlated with fibrinogen and D-Dimer levels. According to COVID-19 severity, only platelet count was significantly higher in patients with severe disease, who needed C-PAP or tracheal intubation, as compared to those with less severe disease. No patients had VTE recurrence or bleeding complications at the time of follow up. Summary/Conclusion: Our data suggest persistence of pro-coagulant and inflammatory laboratory patterns in a proportion of patients with COVID-19-related VTE, in spite of anticoagulant therapy. Further studies are needed to establish on whether monitoring these parameters may help in the decision on the duration of anticoagulation in this condition.

Annual review of venous thromboembolism in 2021

In the past year, significant progress has been made in the field of venous thromboembolism (VTE) including risk assessment and anticoagulation prevention, diagnostic strategies and model exploration, new drug development and disease management. Particularly, major breakthroughs have been made in the prevention of VTE with FXI inhibitors and the prevention of novel coronavirus pneumonia with coagulation alterations and anticoagulation interventions. Here, we reviewed the progress and achievements in the field of VTE in the past year, aiming to provide evidence and ideas for the diagnosis, treatment and future studies of VTE.

Acroyschemia in a Male Patient with Covid 19: A Case Report

Coronavirus disease-19 (COVID-19) which is an enveloped RNA beta coronavirus. First emerged in December 2019 in China and rapidly spread worldwide. Although various studies have reported that COVID-19 is associated with a hypercoagulable state and thrombotic complications in critically ill patients. The current SARS-CoV-2 pandemic is causing high morbidity and mortality, its main affectation is at the pulmonary level, however, the virus also affects the cardiovascular system. The vasculature is affected directly by the virus and indirectly by a storm of inflammatory cytokines, causing an excessive inflammatory process, platelet activation, endothelial dysfunction and stasis. Biochemically it manifests itself with altered coagulation parameters, increased levels of D-dimer and Ferritin, which are the most common laboratory abnormalities related to mortality. In our investigation, acroischemic lesions were found in critically ill patients with severe limb ischemia.

MICROCIRCULATION AND SARS-COV-2 INFECTION: A FOLLOW UP STUDY

Objective: SARS-CoV2 infection can lead to several clinical scenarios, named COVID-19, ranging from mild manifestations to acute respiratory distress syndrome (ARDS), coagulation alterations and endothelial dysfunction. The functional impairment of the microcirculation seems play a key role in the pathophysiology and clinical consequences of COVID-19. However, to date there is no evidence of structural microvascular damage related to COVID-19. Design and method: The aim of this study is to investigate microvascular alterations by adaptive optics and vide-ocapillaroscopy in patients recently admitted for COVID-19 and re-evaluated one year later. Methods: We enrolled 153 patients admitted between February and April 2020 at the Hospital of Montichiari (Brescia) and at the Internal Medicine Department of ASST Spedali Civili - University of Brescia for respiratory failure due to SARS-CoV2-related interstitial pneumonia. Patients were evaluated two months after nalysedsation and after one year. All patients underwent a venous blood sampling for hematochemical tests, evaluation of retinal arteriolar morphology by adaptive optics, assessment of basal and total capillary density (BCD and TCD respectively) at the dorsum of the fourth finger of the non-dominant hand by videocapillaroscopy. Results: Fifty patients with completed follow-up were nalysed. An increase of internal lumen (93.8 ± 13.3 vs. 97.3 ± 14.2 micron, p &lt; 0.001) and a reduction of wall to lumen ratio (WLR 0.30 ± 0.03 vs. 0.27 ± 0.03, p &lt; 0.001) were observed at the follow up visit after one year (Figure). No significant differences were observed in BCD in the dorsum of the finger after one year, whereas a significant reduction in TCD was observed (p &lt; 0.001). Microvascular changes were independent of body mass index and the presence of hypertension or diabetes mellitus. Conclusions: Preliminary data from this study show that patients with SARS-Cov2 infection present an improvement of microvascular structure after one year from the disease, such as a reduction in WLR of retinal arterioles. This suggests that COVID19 might induce structural alterations in the microcirculation which contribute to vascular damage. These changes do not seem to be influenced by the weight, presence of hypertension or diabetes.

Machine learning-based model for prediction of clinical deterioration in hospitalized patients by COVID 19

Despite the publication of great number of tools to aid decisions in COVID-19 patients, there is a lack of good instruments to predict clinical deterioration. COVID19-Osakidetza is a prospective cohort study recruiting COVID-19 patients. We collected information from baseline to discharge on: sociodemographic characteristics, comorbidities and associated medications, vital signs, treatment received and lab test results. Outcome was need for intensive ventilatory support (with at least standard high-flow oxygen face mask with a reservoir bag for at least 6 h and need for more intensive therapy afterwards or Optiflow high-flow nasal cannula or noninvasive or invasive mechanical ventilation) and/or admission to a critical care unit and/or death during hospitalization. We developed a Catboost model summarizing the findings using Shapley Additive Explanations. Performance of the model was assessed using area under the receiver operating characteristic and prediction recall curves (AUROC and AUPRC respectively) and calibrated using the Hosmer–Lemeshow test. Overall, 1568 patients were included in the derivation cohort and 956 in the (external) validation cohort. The percentages of patients who reached the composite endpoint were 23.3% vs 20% respectively. The strongest predictors of clinical deterioration were arterial blood oxygen pressure, followed by age, levels of several markers of inflammation (procalcitonin, LDH, CRP) and alterations in blood count and coagulation. Some medications, namely, ATC AO2 (antiacids) and N05 (neuroleptics) were also among the group of main predictors, together with C03 (diuretics). In the validation set, the CatBoost AUROC was 0.79, AUPRC 0.21 and Hosmer–Lemeshow test statistic 0.36. We present a machine learning-based prediction model with excellent performance properties to implement in EHRs. Our main goal was to predict progression to a score of 5 or higher on the WHO Clinical Progression Scale before patients required mechanical ventilation. Future steps are to externally validate the model in other settings and in a cohort from a different period and to apply the algorithm in clinical practice.Registration: ClinicalTrials.gov Identifier: NCT04463706.

The serum of COVID-19 asymptomatic patients up-regulates proteins related to endothelial dysfunction and viral response in circulating angiogenic cells ex-vivo

BackgroundSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has already caused 6 million deaths worldwide. While asymptomatic individuals are responsible of many potential transmissions, the difficulty to identify and isolate them at the high peak of infection constitutes still a real challenge. Moreover, SARS-CoV-2 provokes severe vascular damage and thromboembolic events in critical COVID-19 patients, deriving in many related deaths and long-hauler symptoms. Understanding how these processes are triggered as well as the potential long-term sequelae, even in asymptomatic individuals, becomes essential.MethodsWe have evaluated, by application of a proteomics-based quantitative approach, the effect of serum from COVID-19 asymptomatic individuals over circulating angiogenic cells (CACs). Healthy CACs were incubated ex-vivo with the serum of either COVID-19 negative (PCR −/IgG −, n:8) or COVID-19 positive asymptomatic donors, at different infective stages: PCR +/IgG − (n:8) and PCR −/IgG + (n:8). Also, a label free quantitative approach was applied to identify and quantify protein differences between these serums. Finally, machine learning algorithms were applied to validate the differential protein patterns in CACs.ResultsOur results confirmed that SARS-CoV-2 promotes changes at the protein level in the serum of infected asymptomatic individuals, mainly correlated with altered coagulation and inflammatory processes (Fibrinogen, Von Willebrand Factor, Thrombospondin-1). At the cellular level, proteins like ICAM-1, TLR2 or Ezrin/Radixin were only up-regulated in CACs treated with the serum of asymptomatic patients at the highest peak of infection (PCR + /IgG −), but not with the serum of PCR −/IgG + individuals. Several proteins stood out as significantly discriminating markers in CACs in response to PCR or IgG + serums. Many of these proteins particiArticle title: Kindly check and confirm the edit made in the articletitle.pate in the initial endothelial response against the virus.ConclusionsThe ex vivo incubation of CACs with the serum of asymptomatic COVID-19 donors at different stages of infection promoted protein changes representative of the endothelial dysfunction and inflammatory response after viral infection, together with activation of the coagulation process. The current approach constitutes an optimal model to study the response of vascular cells to SARS-CoV-2 infection, and an alternative platform to test potential inhibitors targeting either the virus entry pathway or the immune responses following SARS-CoV-2 infection.

Effects of Gymnema inodorum Leaf Extract on the Alteration of Blood Coagulation Parameters and Platelet Count in Plasmodium berghei-Infected Mice.

Malaria remains highly prevalent and one of the major causes of morbidity and mortality in tropical and subtropical regions. Alteration of blood coagulation and platelets has played an important role and attributed to increased morbidity in malaria. Hence, this study was performed to investigate the efficacy of Gymnema inodorum leaf extract on Plasmodium berghei-induced alteration of blood coagulation parameters and platelet numbers in mice. Groups of ICR mice were inoculated with 1 × 107 parasitized red blood cells of P. berghei ANKA (PbANKA) and given orally by gavage with 100, 250, and 500 mg/kg of G. inodorum leaf extract (GIE). Chloroquine (10 mg/kg) was used as a positive control. Platelet count and blood coagulation parameters were measured. The results showed that PbANKA induced thrombocytopenia in mice as indicated by markedly decreased platelet count. Decreased platelet count had a negative correlation with the degree of parasitemia with R2 value of 0.6668. Moreover, significantly (p < 0.05) shortened activated partial thromboplastin time was found in PbANKA-infected group, while prothrombin time and thrombin time were still normal. GIE gave significantly (p < 0.05) good results with respect to platelet count, compared with the results obtained from positive and healthy controls. Additionally, GIE reversed the alteration of blood coagulation parameters when compared to untreated mice. The highest efficacy of GIE was observed at a dose of 500 mg/kg. It was concluded that GIE exerted a protective effect on thrombocytopenia and altered blood coagulation parameters induced by PbANKA infection in mice. This plant may be a future candidate for alternative antimalarial development.

Association of MTHFR rs1801133 and homocysteine with Legg\u2013Calv\xe9\u2013Perthes disease in Mexican patients

BackgroundLegg–Calvé–Perthes disease (LCPD) is the avascular osteonecrosis of the proximal femoral epiphysis. It is a rare disease of unclear etiology in children, although alterations in coagulation or the collagen gene have been described and could be associated with its etiology. Our objective was to evaluate the following alterations: COL1A1 (rs1107946, rs2412298), COL2A1 (rs121912891 and rs387106558), MTHFR rs1801133, CBS rs115742905, and PT rs1799963 and their relationship with LCPD.MethodsDNA was obtained and genotyped by real-time PCR with TaqMan probes. Prothrombin (FII) and homocysteine (Hcy) were determined by a coagulometric method. The variables were described as mean and standard deviation or percentages, and genotypic and allelic distributions were analyzed using the Student's t-test. The Hardy–Weinberg equilibrium and OR were also used.ResultsWe studied 23 patients with LCPD and 46 controls. We did not find any association of the MTHFR, CBS, PT, COL1A1, and COL2A1 genetic variants with LCPD. However, when adjusting the data with the Hcy values for the MTHFR C677T polymorphism, the C/C genotypes showed an association with the recessive model (p = 0.038), with susceptibility to LCPD.ConclusionNo association was found with the CBS, PT, COL1A1, and COL2A1 genes. Nevertheless, our results suggest a significant link between moderately elevated Hcy levels and the MTHFR C677T polymorphism in a cohort of Mexican children with LCPD.

Coagulation abnormalities in patients with COVID-19 on venovenous ECLS increased risk for technical complications and support times but had no impact on survival.

BackgroundPatients with severe coronavirus disease‐19 (COVID‐19)‐associated acute respiratory distress on venovenous extracorporeal lung support (V‐V ECLS) showed a high incidence of vascular as well as ECLS‐related thrombotic complications. The latter may influence the outcome of the patients.MethodsThis is a retrospective monocentric study on prospectively collected data of technical complications including 69 adult COVID‐19 patients on V‐V ECLS (ECLS Registry, March 2020 until April 2021) without and with system exchanges. Alterations in ECLS‐specific data, hemolysis, coagulation, and hemostasis parameters were analyzed.ResultsEvery second COVID‐19 patient on V‐V ECLS developed technical complications. Optimized ECLS management at our ECLS center reduced cases of acute clot formation (pump head thrombosis, acute oxygenator thrombosis) (17%), and allowed early identification of progressive clotting processes (worsened gas transfer, coagulation disorder) (14%, 54%) with a significant overhang of hyperfibrinolysis (37%). Although COVID‐19 disease and technical complications caused the prolonged length of stay at the intensive care unit and ECLS support times, the proportion of successful weaning and survival rates were comparable with patients without system exchange.ConclusionThe survival of ECLS patients with COVID‐19 was independent of the requirement for system exchange due to technical‐induced coagulation disorders. Close monitoring for circuit clotting is mandatory in COVID‐19 patients and is one prerequisite for successful organ support in these difficult patients.

Coagulation imbalance is associated with hepatic fibrosis and vascular complications in patients with type2 diabetes and NAFLD

Introduction: Non-alcoholic fatty liver disease (NAFLD) and type 2 diabetes mellitus (T2DM) are characterized by a pro-coagulant state and vascular complications. Aim: to evaluate in patients with T2DM and NAFLD if alterations in coagulation are associated with hepatic fibrosis and vascular complications.