Procoagulatory state in inflammatory bowel diseases is promoted by impaired intestinal barrier function

Inflammatory and immune mediated disorders are risk factors for arterial and venous thromboembolism. Inflammatory bowel diseases (IBD) confer an even greater risk of thromboembolic events than other inflammatory conditions. It has been shown that IBD patients display defective intestinal barrier functions. Thus, pathogen-associated molecular patterns (PAMPs) coming from the intestinal bacterial burden might reach systemic circulation and activate innate immunity receptors on endothelial cells and platelets, promoting a procoagulative state. Aim of the study was to test this hypothesis, correlating the presence of circulating PAMPs with the activation of innate immune system and the activation of the coagulatory cascade in IBD patients. Specifically, we studied lipopolysaccharide (LPS), Toll-like receptor (TLR) 2, TLR4, and markers of activated coagulation (i.e., D-Dimer and prothrombin fragment F1 + 2) in the serum and plasma of IBD patients. We found that LPS levels are increased in IBD and correlate with TLR4 concentrations; although a mild correlation between LPS and CRP levels was detected, clinical disease activity does not appear to influence circulating LPS. Instead, serum LPS correlates with both D-Dimer and F1 + 2 measurements. Taken together, our data support the role of an impairment of intestinal barrier in triggering the activation of the coagulatory cascade in IBD

Evaluation of an automated chromogenic assay for Factor VIII clotting activity measurement in patients affected by haemophilia A

Introduction: The original one-stage clotting assay is still the most widely used method to measure Factor VIII clotting activity (FVIII:C) in patients with haemophilia A (HA), although the use of chromogenic assays is increasing significantly. Aim: Evaluation of the analytical performance and diagnostic accuracy of BIOPHEN\u2122 FVIII:C (HYPHEN BioMed, Neuville-sur-Oise, France) assay on Sysmex CS-2400 (Sysmex, Kobe, Japan) analyser. Methods: Sixty patients with haemophilia A (HA; any severity) and 120 healthy Italian subjects were included. All the assays were performed on citrate platelet-poor plasmas stored at 1280\ub0C. Chromogenic BIOPHEN\u2122 FVIII:C was compared with the one-stage assay using Actin FS and Factor VIII deficient plasma (Siemens Healthcare Diagnostics, Marburg, Germany) on Sysmex CS-2400 and with another chromogenic automated assay (COAMATIC\u2122 Factor VIII, CHROMOGENIX on ACL TOP analyzer; Instrumentation Laboratory, Milan, Italy). Results: Intra-assay and inter-assay coefficient of variation were <6%. Linearity was good up to 1/128 dilution (r = 0.99); mean recovery was 91.7% and limit of detection was 0.2%. BIOPHEN\u2122 FVIII:C assay showed a good correlation and diagnostic agreement with the chromogenic COAMATIC\u2122 assay: the Spearmen's Rank correlation coefficient was 0.98 and the inter-rate agreement K Cohen coefficient was 0.61. The K coefficient was 0.91 when BIOPHEN\u2122 FVIII:C was compared with the historical classification of the patients, demonstrating an optimal diagnostic accuracy in HA. Conclusions: BIOPHEN\u2122 FVIII:C showed good analytical performance and diagnostic accuracy and could be considered suitable for the introduction in routine analytical panel of coagulation for the diagnosis of HA patients

Inflammatory and prothrombotic parameters in normotensive non-diabetic obese women : effect of weight loss obtained by gastric banding

Hypertension and diabetes are known risk factors for obesity-related thrombosis, but several studies have shown that obesity is characterised by a potentially prothrombotic inflammatory state because of activated coagulation and impaired fibrinolysis. In order to verify if obese patients-unaffected by hypertension, diabetes, dyslipidemia, cigarette smoking or inflammatory diseases-show increased prothrombotic markers and whether the weight loss induced by gastric banding normalises such parameters. Plasma levels of C reactive protein (CRP), fibrinogen, plasminogen activator inhibitor-1 (PAI-1), von Willebrand factor (VWF) and factor VII (FVII) were measured in 25 women with isolated obesity prior to, as well as 3, 6 and 12\ua0months subsequent to gastric banding. Twenty-five healthy women served as a baseline control group. The obese women had higher CRP (p\ua0=\ua00.0001), fibrinogen (p\ua0=\ua00.014), PAI-1 (p\ua0=\ua00.003), VWF (p\ua0=\ua00.004) and FVII levels (p\ua0=\ua00.0001) than the normal controls, and their body mass index (BMI) positively correlated with CRP (r\ua0=\ua00.462, p\ua0=\ua00.02), fibrinogen (r\ua0=\ua00.426, p\ua0=\ua00.04) and PAI-1 (r\ua0=\ua00.468, p\ua0=\ua00.02). Twelve months after gastric banding, the median BMI had decreased from 40.0 to 34.9 (p\ua0=\ua00.0001); CRP from 4.18 to 1.69\ua0\u3bcg/ml (p\ua0=\ua00.01); fibrinogen from 389 to 318\ua0mg/dl (p\ua0=\ua00.0001); PAI-1 from 32.1 to 12.0\ua0UI/ml (p\ua0=\ua00.003); VWF from 144 to 120% (p\ua0=\ua00.0001); and FVII from 134 to 112% (p\ua0=\ua00.002). Even in the absence of major cardiovascular risk factors, obese patients are characterised by a prothrombotic state. The weight loss induced by gastric banding decreases the parameters of inflammation, coagulation and impaired fibrinolysis, thus potentially reducing the thrombotic risk

Antibodies to tissue-type plasminogen activator (t-PA) in patients with inflammatory bowel disease : high prevalence, interactions with functional domains of t-PA and possible implications in thrombosis

Background: Patients with inflammatory bowel disease (IBD) have an increased prevalence of thromboembolic events. The pathogenetic mechanisms of these events include reduced fibrinolysis, which may be caused by antibodies to tissue-type plasminogen activator (t-PA). Objectives: To evaluate anti-t-PA antibodies in patients with IBD, considering clinical, biochemical and functional characteristics. Patients and methods: We immunoenzymatically measured anti-t-PA antibodies in plasma from 97 consecutive IBD patients and 97 ageand sex-matched healthy controls. We also assessed the antibody interactions with different epitopes of t-PA, the antibody inhibition on t-PA activity and the correlations with clinical features and other serum antibodies. Results: IBD patients had higher median anti-t-PA antibody levels (5.4 U mL(-1) vs. 4.0 U mL(-1); P < 0.0001): 18 patients were above the 95th percentile of the controls (OR 5.3; 95% CI 1.7-16.3; P < 0.003), and the six with a history of thrombosis tended to have high levels (6.9 U mL(-1)). Anti-t-PA antibody levels did not correlate with IBD type, activity, location or treatment, or with age, sex, acute-phase reactants or other antibodies. The anti-t-PA antibodies were frequently IgG1 and bound t-PA in fluid phase; they recognized the catalytic domain in 10 patients and the kringle-2 domain in six. The IgG fraction from the three patients with the highest anti-t-PA levels slightly reduced t-PA activity in vitro. Conclusions: The prevalence of anti-t-PA antibodies is high in IBD patients. By binding the catalytic or kringle-2 domains of t-PA, these antibodies could lead to hypofibrinolysis and contribute to the prothrombotic state of IBD

The ADAMTS13-von Willebrand factor axis in COVID-19 patients

Background: Severe coronavirus disease 2019 (COVID-19) is characterized by an increased risk of thromboembolic events, with evidence of microthrombosis in the lungs of deceased patients. Objectives: To investigate the mechanism of microthrombosis in COVID-19 progression. Patients/Methods: We assessed von Willebrand factor (VWF) antigen (VWF:Ag), VWF ristocetin-cofactor (VWF:RCo), VWF multimers, VWF propeptide (VWFpp), and ADAMTS13 activity in a cross-sectional study of 50 patients stratified according to their admission to three different intensity of care units: low (requiring high-flow nasal cannula oxygenation, n&nbsp;=&nbsp;14), intermediate (requiring continuous positive airway pressure devices, n&nbsp;=&nbsp;17), and high (requiring mechanical ventilation, n&nbsp;=&nbsp;19). Results: Median VWF:Ag, VWF:RCo, and VWFpp levels were markedly elevated in COVID-19 patients and increased with intensity of care, with VWF:Ag being 268, 386, and 476&nbsp;IU/dL; VWF:RCo 216, 334, and 388&nbsp;IU/dL; and VWFpp 156, 172, and 192&nbsp;IU/dL in patients at low, intermediate, and high intensity of care, respectively. Conversely, the high-to-low molecular-weight VWF multimers ratios progressively decreased with increasing intensity of care, as well as median ADAMTS13 activity levels, which ranged from 82&nbsp;IU/dL for patients at low intensity of care to 62 and 55&nbsp;IU/dL for those at intermediate and high intensity of care. Conclusions: We found a significant alteration of the VWF-ADAMTS13 axis in COVID-19 patients, with an elevated VWF:Ag to ADAMTS13 activity ratio that was strongly associated with disease severity. Such an imbalance enhances the hypercoagulable state of COVID-19 patients and their risk of microthrombosis

Stress neuropeptide levels in adults with chest pain due to coronary artery disease: potential implications for clinical assessment

: Substance P (SP) and neuropeptide Y (NPY) are neuropeptides involved in nociception. The study of biochemical markers of pain in communicating critically ill coronary patients may provide insight for pain assessment and management in critical care. Purpose of the study was to to explore potential associations between plasma neuropeptide levels and reported pain intensity in coronary critical care adults, in order to test the reliability of SP measurements for objective pain assessment in critical care