Hemostasis Disturbances Research Articles

Fibrin clot permeability (Ks) in patients on left ventricular assist device

Patients on left ventricular assist devices (LVAD) are prone to excessive hemostasis disturbances due to permanent contact of artificial pump surfaces with blood components. We aimed to investigate if fibrin clot permeability is altered in patients on long-term continuous-flow LVAD therapy and if the clot permeability is associated with clinical characteristics and adverse events. We investigated 85 end-stage heart failure patients (90.6% men, age 48.6–63.8 years) scheduled for continuous flow long-term LVAD support according to current clinical indications. The patients were assessed periodically: prior to LVAD implantation (T1), 3–6 months (T2) after LVAD implantation, 6–12 months after (T3) and then every 6 months. We tested the first three blood samples (T1-T3) and the last available blood sample (T4), but no longer than 5 years after LVAD implantation. We assessed hemostasis parameters (Activated Partial Thromboplastin Time (APTT) Prothrombin Time, Activated Partial Thromboplastin Time, Fibrinogen, d-dimer, Antithrombin, Thrombin Time, Factor VIII, and von Willebrand Factor, aspirin-induced platelet inhibition, adenosine-diphosphate test) changes during the study period. Fibrin Clot Permeability was evaluated using a pressure system and Permeability Coefficient (Ks) was calculated. We observed a decrease in fibrin clot permeability (Ks) between T1, T2, T3 and T4 time periods; P < 0.01 for each comparison. Fibrin clot permeability was negatively correlated with fibrinogen concentration: r = − 0.51, P < 0.001, factor VIII activity r = − 0.42, P < 0.001. There was no association of Ks with age, Left Ventricular Ejection Fraction (LVEF) and medications P > 0.001, however cumulative measurements in patients on aspirin showed shortening of Ks in this group P = 0.0123. Major adverse cardiac and cerebrovascular events (MACCE) occurred in 36.5% patients, bleeding events in 25.9%, Net Adverse Clinical Events (NACE) in 62.4%; 31.7% patients died, and 17.6% underwent transplantation. The transplantation was considered as the endpoint. Discrepancies in Ks were observed between patients with MACCE, bleeding, and NACE, and patients without adverse events. Ks showed a constant trend towards normalization (P < 0.01) only in patients without adverse events. Patients with advanced heart failure have disturbed clot structure. A trend towards normalization of the Ks values is associated with fewer thromboembolic and bleeding complications in this group of patients.

Role of Perturbated Hemostasis in MASLD and Its Correlation with Adipokines.

The prevalence of metabolic dysfunction-associated steatotic liver disease (MASLD) continues to rise, making it one of the most prevalent chronic liver disorders. MASLD encompasses a range of liver pathologies, from simple steatosis to metabolic dysfunction-associated steatohepatitis (MASH) with inflammation, hepatocyte damage, and fibrosis. Interestingly, the liver exhibits close intercommunication with fatty tissue. In fact, adipose tissue could contribute to the etiology and advancement of MASLD, acting as an endocrine organ that releases several hormones and cytokines, with the adipokines assuming a pivotal role. The levels of adipokines in the blood are altered in people with MASLD, and recent research has shed light on the crucial role played by adipokines in regulating energy expenditure, inflammation, and fibrosis in MASLD. However, MASLD disease is a multifaceted condition that affects various aspects of health beyond liver function, including its impact on hemostasis. The alterations in coagulation mechanisms and endothelial and platelet functions may play a role in the increased vulnerability and severity of MASLD. Therefore, more attention is being given to imbalanced adipokines as causative agents in causing disturbances in hemostasis in MASLD. Metabolic inflammation and hepatic injury are fundamental components of MASLD, and the interrelation between these biological components and the hemostasis pathway is delineated by reciprocal influences, as well as the induction of alterations. Adipokines have the potential to serve as the shared elements within this complex interrelationship. The objective of this review is to thoroughly examine the existing scientific knowledge on the impairment of hemostasis in MASLD and its connection with adipokines, with the aim of enhancing our comprehension of the disease.

Update on Anticoagulation Strategies in Patients with ECMO-A Narrative Review.

The use of extracorporeal membrane oxygenation (ECMO) has recently increased exponentially. ECMO has become the preferred mode of organ support in refractory respiratory or circulatory failure. The fragile balance of haemostasis physiology is massively altered by the patient's critical condition and specifically the aetiology of the underlying disease. Furthermore, an application of ECMO conveys another disturbance of haemostasis due to blood-circuit interaction and the presence of an oxygenator. The purpose of this review is to summarise current knowledge on the anticoagulation management in patients undergoing ECMO therapy. The unfractionated heparin modality with monitoring of activated partial thromboplastin tests is considered to be a gold standard for anticoagulation in this specific subgroup of intensive care patients. However, alternative modalities with other agents are comprehensively discussed. Furthermore, other ways of monitoring can represent the actual state of coagulation in a more complex fashion, such as thromboelastometric/graphic methods, and might become more frequent. In conclusion, the coagulation system of patients with ECMO is altered by multiple variables, and there is a significant lack of evidence in this area. Therefore, a highly individualised approach is the best solution today.

Explore the impact of abnormal coagulation test results on pregnancy complications and perinatal outcomes by establishing the trimester-specific reference intervals of singleton and twin pregnancies

BackgroundDuring pregnancy, complex physiological changes take place in the hemostatic system, resulting in a hypercoagulable state. With the established trimester-specific reference intervals (RIs) of the coagulation tests, we investigated the associations between disturbance of hemostasis and adverse pregnant outcomes in a population-based cohort study. MethodsThe first- and third-trimester coagulation tests results were retrieved from 29,328 singleton and 840 twin pregnant women for regular antenatal check-ups from November 30th, 2017 to January 31st, 2021. The trimester-specific RIs for fibrinogen (FIB), prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), d-dimer (DD) were estimated using both the direct observational and the indirect Hoffmann methods. The associations between the coagulation tests and the risks of developing pregnancy complications as well as adverse perinatal outcomes were assessed using the logistic regression analysis. ResultsIncreased FIB, DD and decreased PT, APTT and TT were observed as the gestational age increases in the singleton pregnancy. An enhanced procoagulant state, marked by significant elevation of FIB, DD and reduction of PT, APTT and TT, was observed in the twin pregnancy. The subjects with anormal PT, APTT, TT, DD, tend to have increased risks of developing peri- and postpartum complications such as preterm birth, fetal growth restriction. ConclusionsThe incidence of adverse perinatal outcomes was remarkably associated with the maternal increased levels of FIB, PT, TT, APTT and DD in the third trimester, which may be applied in early identification of women at high risk of adverse outcomes due to coagulopathy.

The correlation between Intra-abdominal Adhesions and body mass index in Women Undergoing elective repeated Cesarean Section

Background: Intra-abdominal adhesion formation is serious post-surgery problem that occurs frequently after caesarean section. Obesity is associated with many hemostasis disturbances, particularly impaired fibrinolysis, which may increase the incidence of intra-abdominal adhesions. Aim of Work: Goal of the work was to correlate between intra-abdominal adhesions and body mass index in women scheduled for a repeated cesarean section. Patients & Methods: This was observational cross-sectional research that was carried out at Obstetrics & Gynecology department, faculty of medicine, Al-Azhar university hospitals during the period between October 2021 and April 2022, and total of 232 pregnant women undergoing repeated cesarean section, were enrolled in our study. They were separated into 2 categories according to the BMI: Category I: BMI < thirty kg/m2 (N = 30) and Category II: BMI ≥ thirty kg/m2 (N = 202); Group IIa (N = 96): BMI thirty – 34.9 kg/m2; Group IIb (N = 81): BMI thirty five – 39.9 kg/m2, and Group IIc (N = 25): BMI ≥ 40 kg/m2. Results: no statistically significant variation was discovered among categories regarding preoperative Hgb. However, group II was associated with more significant blood loss compared to group I. Furthermore, the incidence of intraoperative visceral injuries was greater in category II compared to category I. Conclusion: in women undergoing cesarean section, there was a significant association between body mass index and formation of intra-abdominal adhesion with no effect on scar thickness, more frequent depressed scar and dense adhesions, larger adhesion size, longer operation duration and no effect on intraoperative bleeding.

Label-Free Mass Spectrometry-Based Quantitative Proteomics to Evaluate the Effects of the Calcium-Sensing Receptor Agonist Cinacalcet on Protein Expression in Rat Brains and Livers.

BackgroundCinacalcet is a calcium-sensing receptor agonist that is clinically approved for the treatment of secondary hyperparathyroidism in chronic kidney disease and hypercalcemia in patients with parathyroid carcinoma. This study aimed to use quantitative mass spectrometry-based label-free proteomics to evaluate the effects of cinacalcet on protein expression in rat brains and livers.Material/MethodsWe randomly assigned 18 Wistar rats to 2 groups: an untreated control group (n=6) and a group treated with cinacalcet at a dose corresponding to the maximum dose used in humans (2 mg/kg/body weight, 5 days/week) divided into 7-day (n=6) and 21-day (n=6) treatment subgroups. A mass-spectrometry-based label-free quantitative proteomics approach using peptides peak area calculation was used to evaluate the changes in protein expression in examined tissues. Bioinformatics analysis of quantitative proteomics data was done using MaxQuant and Perseus environment.ResultsNo changes in protein expression were revealed in the 7-day treatment subgroup. We detected 10 upregulated and 3 downregulated proteins in the liver and 1 upregulated protein in the brain in the 21-day treatment subgroup compared to the control group. Based on Gene Ontology classification, all identified differentially expressed proteins were indicated as molecular functions involved in the enzyme regulator activity (36%), binding (31%), and catalytic activity (19%).ConclusionsThese findings indicate that long-term cinacalcet therapy can impair phase II of enzymatic detoxication and can cause disturbances in blood hemostasis, lipid metabolism, and inflammatory mediators or contribute to the acceleration of cognitive dysfunction; therefore, appropriate patient monitoring should be considered.

THU519 - Disturbances in sodium and chloride hemostasis predict outcome in stable and critically-ill patients with cirrhosis-not two sides of the same coin

THU519 - Disturbances in sodium and chloride hemostasis predict outcome in stable and critically-ill patients with cirrhosis-not two sides of the same coin

The Role of Vascular-Platelet Hemostasis Disturbances in the Genesis of Respiratory Failure in Patients with COVID-19

The objective: to study the relationship between markers of imbalance in the platelet-vascular hemostasis system and lung damage and disease outcome in patients with COVID-19.Subjects and Methods: the study included 225 patients with the confirmed diagnosis of moderate, severe and extremely severe COVID-19. In all patients, lung damage was confirmed using computed and X-ray diagnostic methods. We studied the concentration of von Willebrand factor (vWF), activity of metalloproteinase ADAMTS-13, concentration of homocysteine, endothelin-1, thrombomodulin and the number of platelets and changes in these parameters on the 1st, 3–5th and 7–10th days from the admission to the intensive care unit (ICU). In deceased patients (n = 106), lung specimens were collected to assess the nature of morphological changes.Results. It was determined that by the 7–10th days with a lethal outcome of the disease, there was a statistically significant increase in the concentration of vWF by 22.2% (p = 0.0225), the vWF/ADAMTS-13 ratio by 2 times (p = 0.0408) and decrease in the platelet count by 52% (p = 0.0008) versus the initial stage of the study. In all the deceased, the morphological picture of lung tissue damage was characterized by destruction of alveolocytes, proliferation of connective tissue, cellular infiltration, plethora of capillaries, parietal erythrocyte thrombi in the lumen of capillaries, arterioles and venules, and obstructive erythrocyte thrombi in the lumen of vessels of lung microcirculation.Conclusion. The study showed that a possible cause of thrombosis in the lumen of vessels of lung microcirculation in the case of a lethal outcome of COVID-19 might be a primary imbalance in the vascular-platelet hemostasis characterized by a significant increase in the concentration of the vWF factor, the vWF/ADAMTS-13 ratio and a decrease in the number (consumption) of platelets by the 7–10th day of the disease versus the initial stage of the study.

Hemostasis Disturbances in Continuous-Flow Left Ventricular Assist Device (CF-LVAD) Patients—Rationale and Study Design

Left ventricular assist devices are a treatment option for end-stage heart failure patients. Despite advancing technologies, bleeding and thromboembolic events strongly decrease the survival and the quality of life of these patients. Little is known about prognostic factors determining these adverse events in this group of patients. Therefore, we plan to investigate 90 consecutive left ventricular assist device (LVAD) patients and study in vitro fibrin clot properties (clot lysis time, clot permeability, fibrin ultrastructure using a scanning electron microscope) and the calibrated automated thrombogram in addition to the von Willebrand factor antigen, fibrinogen, D-dimer, prothrombin time/international normalized ratio (PT/INR), and activated partial thromboplastin time (APTT) to identify prognostic factors of adverse outcomes during the course of therapy. We plan to assess the hemostasis system at four different time points, i.e., before LVAD implantation, 3–4 months after LVAD implantation, 6–12 months after LVAD implantation, and at the end of the study (at 5 years or at the time of the adverse event). Adverse outcomes were defined as bleeding events (bleeding in general or in the following subtypes: severe bleeding, fatal bleeding, gastrointestinal bleeding, intracranial bleeding), thromboembolic events (stroke or transient ischemic attack, pump thrombosis, including thrombosis within the pump or its inflow or outflow conduits, arterial peripheral thromboembolism), and death.

The importance of α1AMP-activated protein kinase (α1AMPK) in hemostatic dysfunctions induced by sepsis

Sepsis is a pathological circumstance whose outcome is associated with organ dysfunction and death. Vascular hyperpermeability and hemostasis impairment both play a key role in the onset of organ failure. Our previous work highlighted the protective role of α1AMPK against sepsis-induced hyperpermeability. However, the impact of α1AMPK on hemostasis has never been investigated in this condition. This study aims to characterize the interaction of α1AMPK with hemostasis in sepsis. α1AMPK wild type (WT) and knockout (KO) mice were challenged with intraperitoneal injection of lipopolysaccharide (LPS) from Escherichia coli O55B5 (10 mg/kg) during 6 and 18 hours. Plasmatic markers of coagulation, NETosis and platelet activation were measured by ELISA. To identify cell type responsible for the dysregulation, same analyses were conducted on endothelial-, leucocyte- or platelet- specific α1AMPK knockout mouse models. Hemostasis was altered at 6 hours and 18 hours after LPS administration, in both α1AMPK WT and KO mice. Thrombin generation, reflected by thrombin-antithrombin complex formation, was significantly increased in α1AMPK KO mice compared to WT mice, after 18 hours of treatment with LPS. In contrast, fibrinolysis was more inhibited in α1AMPK KO than in WT mice, with a higher level of plasminogen activator inhibitor-1 measured at 6 and 18 hours, while tissue plasminogen activator was not different. This exacerbated procoagulant phenotype in α1AMPK KO mice was associated with an increase of sCD62P in plasma, reflecting an overactivation of platelets under septic conditions. Septic α1AMPK KO mice also showed an increase of Neutrophil Elastase and plasma DNA with no variation of MPO-DNA complexes, suggesting that NETosis was not altered but that exacerbated cellular damages occurred in these mice compared to WT animals. Platelet- and leucocyte- specific α1AMPK knockout mouse models did not recapitulate the phenotype observed in total α1AMPK knockout mice after LPS administration. However, endothelial specific α1AMPK knockout mice showed an increase of thrombin generation with a tendency to higher platelet activation. Altogether, these results demonstrate that α1AMPK plays a crucial role in the regulation of hemostasis disturbances during sepsis. While α1AMPK from platelets and leucocytes does not contribute to the septic damage, endothelialα1AMPK might be a brake for hemostatic dysregulation.

The importance of alpha-1 AMP-activated protein kinase (α1AMPK) in hemostatic dysfunctions induced by sepsis

Sepsis is a pathological circumstance whose outcome is associated with organ dysfunction and death. Vascular hyperpermeability and hemostasis impairment, both play a key role in the onset of organ failure. Our previous work highlighted the protective role of α1AMPK against sepsis-induced hyperpermeability. However, the impact of α1AMPK on hemostasis has never been investigated in this condition. This study aims to characterize the interaction of α1AMPK with hemostasis in sepsis. α1AMPK wild type (WT) and knockout (KO) mice were challenged with intraperitoneal injection of lipopolysaccharide (LPS) from Escherichia coli O55B5 (10 mg.kg) during 6 and 18 hours. Plasmatic markers of coagulation, NETosis and platelet activation were measured by ELISA. In order to identify the origin of the dysregulation, same analyses were conducted on endothelial, leucocyte or platelet specific deletion model. In both α1AMPK WT and KO mice, hemostasis was altered at 6 hours and 18 hours after LPS administration. Thrombin generation, reflected by thrombin-antithrombin complex formation, was significantly increased in α1AMPK KO mice compared with WT mice, after 18 hours of treatment by LPS. In contrast, fibrinolysis was more inhibited in α1AMPK KO than in WT mice, with a higher level of plasminogen activator inhibitor-1 measured at 6 and 18 hours, while tissue plasminogen activator was not different. This exacerbated procoagulant phenotype in α1AMPK KO mice was associated with an increase in sCD62P in plasma, showing an overactivation of platelets under septic conditions. Septic α1AMPK KO mice showed an increase of Neutrophil Elastase and plasmatic DNA with no variation of MPO-DNA complex suggesting a stronger cellular damage than WT mice with no increase of NETosis. Platelet and leucocyte specific models did not have the phenotype observed in the total deletion model after LPS administration. However, endothelial specific model showed an increase of thrombin generation with a tendency of more platelet activation. Altogether, these results demonstrate that α1AMPK plays an important role in the regulation of hemostasis disturbances during sepsis. Specific model showed that α1AMPK from platelet and leucocyte does not contribute directly to the septic damage but endothelial α1AMPK seems to be important to contain hemostasis dysregulation.

The Bioflavonoids Rutin and Rutin Succinate Neutralize the Toxins of B. jararaca Venom and Inhibit its Lethality.

The venom of the Brazilian pit viper Bothrops jararaca (BjV) is a complex mixture of molecules, and snake venom metalloproteinases (SVMP) and serine proteinases (SVSP) are the most abundant protein families found therein. Toxins present in BjV trigger most of the deleterious disturbances in hemostasis observed in snakebites, i.e., thrombocytopenia, hypofibrinogenemia and bleedings. The treatment of patients bitten by snakes still poses challenges and the bioflavonoid rutin has already been shown to improve hemostasis in an experimental model of snakebite envenomation. However, rutin is poorly soluble in water; in this study, it was succinylated to generate its water-soluble form, rutin succinate (RS), which was analyzed comparatively regarding the chemical structure and characteristic features of rutin. Biological activities of rutin and RS were compared on hemostatic parameters, and against toxic activities of crude BjV in vitro. In vivo, C57BL/6 mice were injected i.p. with either BjV alone or pre-incubated with rutin, RS or 1,10-phenanthroline (o-phe, an SVMP inhibitor), and the survival rates and hemostatic parameters were analyzed 48 h after envenomation. RS showed the characteristic activities described for rutin – i.e., antioxidant and inhibitor of protein disulfide isomerase – but also prolonged the clotting time of fibrinogen and plasma in vitro. Differently from rutin, RS inhibited typical proteolytic activities of SVMP, as well as the coagulant activity of BjV. Importantly, both rutin and RS completely abrogated the lethal activity of BjV, in the same degree as o-phe. BjV induced hemorrhages, falls in RBC counts, thrombocytopenia and hypofibrinogenemia in mice. Rutin and RS also improved the recovery of platelet counts and fibrinogen levels, and the development of hemorrhages was totally blocked in mice injected with BjV incubated with RS. In conclusion, RS has anticoagulant properties and is a novel SVMP inhibitor. Rutin and RS showed different mechanisms of action on hemostasis. Only RS inhibited directly BjV biological activities, even though both flavonoids neutralized B. jararaca toxicity in vivo. Our results showed clearly that rutin and RS show a great potential to be used as therapeutic compounds for snakebite envenomation.

Analysis of Disturbances in Secondary Haemostasis in Patients with Tick Bite

Analysis of Disturbances in Secondary Haemostasis in Patients with Tick Bite

Элементный состав плодов наиболее распространенных растений семейства Vacciniaceae

Introduction. Disturbances of trace element hemostasis are the causes of trace element imbalance, which often lead to the development of such major human diseases as iron deficiency anemia (IDA) and diabetes mellitus. Aim. Study of the trace element composition of fruits of the Vacciniaceae family plants and determination of their prospects for use in the prevention of trace element imbalance. Materials and methods. The content of 37 elements, including essential (Mn, Fe, Cu, Cr, Co, Zn) and toxic elements (Pb, Hg, As, and Cd), was determined by inductively coupled plasma mass spectroscopy. Results. The content of toxic elements in all the studied samples is below the reference values. The content of essential microelements differs, which must be taken into account in the prevention of specific imbalance of trace elements. Conclusion. The collected raw materials are environmentally safe and can be used for medicinal purposes. The combination of cranberry and blueberry is optimal in terms of manganese and zinc content and can be used to prevent diabetes. From the point of view of the IDA prevention, it is preferable to use blueberry fruits, which have equimolar concentrations of iron and manganese and the maximum content of cobalt.

ATRIAL FIBRILLATION IN PATIENTS WITH CONCOMITANT DIABETES MELLITUS – WHAT DO WE ALREADY KNOW AND WHAT DO WE NEED TO DISCOVER?

Recently published data indicate the prevalence of atrial fibrillation, the most common cardiac arrhythmia worldwide, in up to 20% of the elderly population. This arrhythmia significantly impacts the quality of life by increasing the risk of stroke, thromboembolism, dementia or heart failure, resulting in a substantial increase in the risk of adverse events and all-cause death. On the other hand, diabetes mellitus is the most predominant metabolic disorder on the globe, which incidence is surging annually, currently affecting over 500 million individuals. Patients with coexisting diabetes have a relevantly elevated risk of atrial fibrillation development. This association have not yet been comprehensively elucidated. Nonetheless, it seems to be a multifactorial, complex relationship comprising mechanisms such as oxidative stress, insulin resistance, hemostasis and fibrinolysis disturbances or endothelium dysfunction, which lead to mechanical and electrical left atrial remodeling. Therefore, this study aims to summarize the evidence regarding the relationship linking diabetes mellitus and atrial fibrillation.

The state of hemostasis in perimenopause and hormonal replacement therapy

The interruption in the ovarianfunctioning goes along with the disturbances of vascular-trombocyte hemostasis: intravascular trombocyte activation increases, trombocyte micro- and macro-aggregates appear. Cyclic hormonal replacement therapy with Femoston, which contains estradiol and didrogesterone, helps to normalize the trombocytogram. Besides that, the addition of progestagen didrogesterone increases the effect of estradiol on micro circular hemostasia.

ASPARAGINASE INDUCED SINUS VEIN THROMBOSIS IN AN ADULT YOUNG ALL PATIENT

Asparaginase has a very important role in ALL treatment among increasing of remission rate and duration. Multiple side effects prevent its regulatory use. Asparaginase reduce antithrombin 3, heparin cofactor II, protein C and plasminogen synthesis. By the way, P-selectin, PAI 1, tissue factor activity, and vWF antigen levels increase. Hypofibrinogenemia may be a marker of of hemostasis disturbances and decreased protein synthesis. A 25-year-old male patient with acute T-cell lymphoblastic leukemia diagnosis was initiated on induction chemotherapy according to augmented BFM protocol. After 4 weeks, remission was confirmed. During consolidation with the third dose of standard L-asparaginase of 10000 units, headache, nausea and vomiting started and confusion developed. Biochemical investigations, PT, aPTT were within normal limits. Fibrinogen was 92 mg/dL and D-dimer was high.Contrast-enhanced MRI showed a thrombus occluding. the superior sagittal sinus. He was intubated and followed up in the intensive care unit due to a rapid decline in the Glasgow score and epileptic seizures. Correction of fibrinogen with cryoprecipitate anticoagulation treatment contributed to symptoms improvement. unfortunately, asparaginase was removed from the protocol. During maintenance therapy, he has had severe COVID-19 pneumonia and during this time he did not experience any thrombosis related complications. Asparaginase therapy is associated with low antithrombin and fibrinogen levels and 7% of thrombosis rate. Previous studies showed that, in ALL patients, thrombosis occurred far more frequently during cycles that contained asparaginase than those that did not. Therefore, studies have investigated the role of fresh frozen plasma or cryoprecipitate supplementation to reduce the thrombohemorrhagic risk of therapy. Retrospective studies suggest antithrombin concentrates may have a beneficial effect on the outcomes of adults treated with asparaginase for ALL.

The main pathogenetic mechanisms of hypercoagulation in diabetes and the possibility of its drug correction

Disorders in the blood coagulation system play an important role in the development of cardiovascular pathology in diabetes. Factors that cause them are hyperglycemia, insulin deficiency, insulin resistance, dyslipidemia, oxidative stress. The most significant changes are observed in the vascular-platelet link of hemostasis. Diabetes is characterized by morphological and functional changes in the endothelium of blood vessels. The activity of platelets increases, which is manifested by their high level of spontaneous aggregation and increased sensitivity to the action of activating factors. The role in the disturbance of hemostasis is played by increasing the activity of the von Willebrand factor, reflecting damage to endothelial cells. Diabetes is characterized by an increase in the activity of plasma clotting factors (I, II, III, VII, VIII, IX, XI, XII and XIII), activation of the callicrein-kinin system. In some cases, this correlates with the development of complications of diabetes. Characteristic disorders in the coagulation inhibition system are a decrease in the activity of antithrombin III, reduced formation of thrombin-antithrombin complexes, reduction of thrombomodulin and protein C. In diabetes, there is a decrease in fibrinolysis, due to a decrease in the expression of tissue activator plasminogen and an increase in the level of the inhibitor of the activator plasminogen. The possibilities of drug correction of hypercoagulation factors in diabetes are to achieve glycemic control with sugar-reducing drugs and elimination of dyslipidemia through hypolipidemic therapy. The most well-studied sugar-lowering drug that improves the state of the blood clotting system is metformin. The system of hemostasis in diabetic patients is positively affected by statins both due to the direct hypolipidemic effect, and by improving endothelial function and increasing fibrinolysis.

Phosphatidylserine on Blood Cells and Microparticles Contributes to the Hyper-Coagulable State in Diabetic Kidney Disease

Introduction: Patients with diabetes mellitus (DM) are considered to be prothrombotic with major disturbances in hemostasis that are associated with an increased risk of venous thromboembolism, especially when microvascular complications of diabetes mellitus occur, such as diabetic kidney disease (DKD). Additionally, microthrombosis in glomerular capillaries could decrease the kidney blood flow and induce renal insufficiency. However, patients with DKD do not receive routine antithrombotic therapy. Therefore, more study is needed on the mechanisms of coagulation in DKD with the aim of identifying therapeutic targets. Recent studies have shown that phosphatidylserine (PS), a membrane constituent, plays an important role in the thrombosis. However, relatively little is known about the precise role of PS in the prothrombotic state of DKD. Our objectives were to study the effect of increased PS exposure on microparticles (MPs) and the outer membrane of MP-origin cells in DKD patients, and to evaluate its effect on procoagulant activity (PCA).Methods: DKD patients (n = 90) were divided into three groups based on urinary albumin excretion rate (UAER), defined as normoalbuminuria (<30 mg/24 h), microalbuminuria (30-299 mg/24 h), or macroalbuminuria (>300 mg/24 h), and compared to healthy controls (n = 30). Lactadherin was used to quantify PS exposure on MPs and their original cells. Healthy blood cells and human umbilical vein endothelial cells (HUVECs) were treated with 25 mM, 5 mM or 2.5 mM glucose and then PS exposure was measured. PCA of MPs and cells was evaluated by clotting time and purified coagulation complex assays. Fibrin production was determined by turbidity. Phosphatidylserine exposure and fibrin strands were observed using confocal microscopy.Results: Usingflow cytometry, we found that PS+ MPs (derived from platelets, erythrocytes, HUVECs, neutrophils, monocytes and lymphocytes) and blood cells (BCs) were significantly higher in patients than in controls. Furthermore, the number of PS+ MPs and BCs in patients with macroalbuminuria was significantly higher than in patients with normoalbuminuria. Similarly, we observed markedly elevated PS exposure on HUVECs cultured with serum from patients with macro-DKD versus serum from patients with micro-DKD or normo-DKD. In addition, circulating PS+ MPs cooperated with PS+ cells, contributing to markedly shortened coagulation time and dramatically increased FXa/thrombin generation and fibrin formation in each DKD group. Confocal microscopy images demonstrated co-localization of fibrin with PS on ECs. Moreover, blockade of exposed PS on MPs and cells with lactadherin inhibited PCA by approximately 80%. Additionally, both hyperglycemia (25 mmol/L) and hypoglycemia (2.5 mmol/L) induced increased PS exposure on cells compared to normoglycemia (5 mmol/L). Linear regression analysis showed that levels of PS+ BCs and MPs were positively correlated with uric acid and proteinuria, but negatively correlated with GFR.Conclusions: Our results suggested that PS+ MPs and blood cells play a prominent role in inducing the hypercoagulable and prothrombotic state in patients with DKD. The occurrence of abnormal glucose during the hypoglycemic therapy is associated with hypercoagulation in DKD. PS+ cells and MPs likely support thrombosis in renal vascular and result in renal dysfunction and disease progression. Our results demonstrate that PS provides binding sites for FXa and prothrombinase complexes and promotes thrombin formation. Thus, directly targeting FXa and prothrombinase complexes might decrease thrombotic and renal disease risk. The results from this study suggest that future research should focus on discovery and deployment of direct PS inhibitors.Disclosure of Interest: Jialan Shi has a patent for the use of lactadherin as a PS probe. DisclosuresNo relevant conflicts of interest to declare.

Platelet and Haemostasis are the Main Targets in Severe Cases of COVID-19 Infection; a System Biology Study.

Introduction: Many proteomics-based and bioinformatics-based efforts are made to detect the molecular mechanism of COVID-19 infection. Identification of the main protein targets and pathways of severe cases of COVID-19 infection is the aim of this study.Methods:Published differentially expressed proteins were screened and the significant proteins were investigated via protein-protein interaction network using Cytoscape software V. 3.7.2 and STRING database. The studied proteins were assessed via action map analysis to determine the relationship between individual proteins using CluePedia. The related biological terms were investigated using ClueGO and the terms were clustered and discussed.Results:Among the 35 queried proteins, six of them (FGA, FGB, FGG, and FGl1 plus TLN1 and THBS1) were identified as critical proteins. A total of 38 biological terms, clustered in 4 groups, were introduced as the affected terms. “Platelet degranulation” and “hereditary factor I deficiency disease” were introduced as the main class of the terms disturbed by COVID-19 virus.Conclusion:It can be concluded that platelet damage and disturbed haemostasis could be the main targets in severe cases of coronavirus infection. It is vital to follow patients’ condition by examining the introduced critical differentially expressed proteins (DEPs).