Activation Of Blood Coagulation Research Articles

Site-Specific Analysis of Core and Antenna Fucosylation on Serum Glycoproteins.

Fucosylation is an important structural feature of glycans and plays an essential role in the regulation of glycoprotein functions. Fucosylation can be classified into core- (CF) and antenna-fucosylation (AF, also known as (sialyl-) Lewis) based on the location on N-glycans, and they perform distinct biological functions. In this study, core- and antenna-fucosylated N-glycans on human serum glycoproteins that hold great clinical application values were systematically characterized at the site-specific level using StrucGP combined with the recently developed fucosylation assignment method. The results showed that fucosylation was widely distributed on serum glycoproteins, with 50% of fucosylated glycopeptides modified by AF N-glycans, 37% by CF N-glycans, and 13% by dual-fucosylated N-glycans. Interestingly, CF and AF N-glycans preferred to modify different groups of serum glycoproteins with different tissue origins and were involved in distinctive biological processes. Specifically, AF N-glycoproteins are mainly from the liver and participated in complement activation, blood coagulation, and endopeptidase activities, while CF N-glycoproteins originate from diverse tissues and are mainly involved in cell adhesion and signaling transduction. These data further enhanced our understanding of fucosylation on circulation glycoproteins.

The Role of Myeloid Cells in Thromboinflammatory Disease.

Inflammation contributes to the development of thrombosis, but the mechanistic basis for this association remains poorly understood. Innate immune responses and coagulation pathways are activated in parallel following infection or injury, and represent an important host defense mechanism to limit pathogen spread in the bloodstream. However, dysregulated proinflammatory activity is implicated in the progression of venous thromboembolism and arterial thrombosis. In this review, we focus on the role of myeloid cells in propagating thromboinflammation in acute inflammatory conditions, such as sepsis and coronavirus disease 2019 (COVID-19), and chronic inflammatory conditions, such as obesity, atherosclerosis, and inflammatory bowel disease. Myeloid cells are considered key drivers of thromboinflammation via upregulated tissue factor activity, formation of neutrophil extracellular traps (NETs), contact pathway activation, and aberrant coagulation factor-mediated protease-activated receptor (PAR) signaling. We discuss how strategies to target the intersection between myeloid cell-mediated inflammation and activation of blood coagulation represent an exciting new approach to combat immunothrombosis. Specifically, repurposed anti-inflammatory drugs, immunometabolic regulators, and NETosis inhibitors present opportunities that have the potential to dampen immunothrombotic activity without interfering with hemostasis. Such therapies could have far-reaching benefits for patient care across many thromboinflammatory conditions.

The Inhibition of Serine Proteases by Serpins Is Augmented by Negatively Charged Heparin: A Concise Review of Some Clinically Relevant Interactions.

Serine proteases are members of a large family of hydrolytic enzymes in which a particular serine residue in the active site performs an essential role as a nucleophile, which is required for their proteolytic cleavage function. The array of functions performed by serine proteases is vast and includes, among others, the following: (i) the ability to fight infections; (ii) the activation of blood coagulation or blood clot lysis systems; (iii) the activation of digestive enzymes; and (iv) reproduction. Serine protease activity is highly regulated by multiple families of protease inhibitors, known collectively as the SERine Protease INhibitor (SERPIN). The serpins use a conformational change mechanism to inhibit proteases in an irreversible way. The unusual conformational change required for serpin function provides an elegant opportunity for allosteric regulation by the binding of cofactors, of which the most well-studied is heparin. The goal of this review is to discuss some of the clinically relevant serine protease-serpin interactions that may be enhanced by heparin or other negatively charged polysaccharides. The paired serine protease-serpin in the framework of heparin that we review includes the following: thrombin-antithrombin III, plasmin-anti-plasmin, C1 esterase/kallikrein-C1 esterase inhibitor, and furin/TMPRSS2 (serine protease Transmembrane Protease 2)-alpha-1-antitrypsin, with the latter in the context of COVID-19 and prostate cancer.

Transcriptome analysis of cultured human vascular endothelial cells after γ-ray irradiation and correlation analysis with ATP, ADP, and adenosine as secondary soluble factors.

Vascular endothelial cells serve as barriers between blood components and subendothelial tissue and regulate the blood coagulation-fibrinolytic system. Ionizing radiation is a common physical stimulant that induces a bystander effect whereby irradiated cells influence neighboring cells through signalings, including purinergic receptor signaling, activated by adenosine 5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), and adenosine as secondary soluble factors. Human vascular endothelial EA.hy926 cells were cultured and irradiated with γ-rays or treated with ATP, ADP, or adenosine under non-toxic conditions. RNA-seq, gene ontology, and hierarchical clustering analyses were performed. The transcriptome analysis of differentially expressed genes in vascular endothelial cells after γ-ray irradiations suggests that the change of gene expression by γ-irradiation is mediated by ATP and ADP. In addition, the expression and activity of the proteins related to blood coagulation and fibrinolysis systems appear to be secondarily regulated by ATP and ADP in vascular endothelial cells after exposure to γ-irradiation. Although it is unclear whether the changes of the gene expression related to blood coagulation and fibrinolysis systems by γ-irradiation affected the increased hemorrhagic tendency through the exposure to γ-irradiation or the negative feedback to the activated blood coagulation system, the present data indicate that toxicity associated with γ-irradiation involves the dysfunction of vascular endothelial cells related to the blood coagulation-fibrinolytic system, which is mediated by the signalings, including purinergic receptor signaling, activated by ATP and ADP.

Dependence of coagulation activity of chicken blood IN VITRO on temperature

The paper presents the results of examine  the  relationship  between  blood  plasma incubation  temperature  and  the  hemostasis parameters in chickens. The purpose of this research was to study of the blood coagulation  parameters  in  chickens  during  hypo-, normo- and hyperthermia in vitro. In addition, to assess the possibility of using a coagulometer to identify hemostasis activity and the use of hemostatic reactions in chickens as models for human medicine. The Vologda region was where the study was conducted. The coagulogram was examined for the following parameters: thrombin time (TT), prothrombin time (PT), activated partial thromboplastin  time  (APTT),  fibrinogen  activity, and antithrombin III activity (AT-III). It was found that at 46° C activation of blood clotting occurs, manifested as an acceleration of the  TT  by  29%  (13  seconds),  PT  by  72% (111  seconds),  and  decrease  in  the  antithrombin III activity by 6% (1.5 seconds) compared to normothermia. The parameters of  secondary  hemostasis  also  responded  to hypothermia (18 °C) with hypercoagulation. It  was  an  acceleration  of  PT  of  81%  (125 sec),  an  increase  in  fibrinogen  activity  by 77% (84 sec) and a decrease in AT-III activity  by  55%  (14  sec),  with  the  exception  of thrombin time, which was extended by 70% (104  seconds).  There  were  no  significant changes  in  the  APTT  at  different  temperatures.  The  correlation-regression  and  one-factor analysis of variance revealed a significant correlation between the temperature and the TT and the fibrinogen activity, where the coefficients of determination for which were 71%  and  39%,  respectively.  Plasma  hemostasis  activity  measured  on  a  coagulometer (at 37 °C) showed no significant differences from  those  measured  at  43  °C,  except  for prothrombin time, which was 71.51 seconds (47%) less than that measured at 43 °C.

Green Solvent-Based Antifouling Polymer Brushes Demonstrate Excellent Hemocompatibility.

Medical devices are crucial for patient care, yet even the best biomaterials lead to infections and unwanted activation of blood coagulation, potentially being life-threatening. While hydrophilic polymer brushes are the best coatings to mitigate these issues, their reliance on fossil raw materials underscores the urgency of bio-based alternatives. In this work, we introduce polymer brushes of a green solvent-based monomer, prohibiting protein adsorption, bacterial colonization, and blood clot formation at the same level as fossil-based polymer brushes. The polymer brushes are composed of N,N-dimethyl lactamide acrylate (DMLA), can be polymerized in a controlled manner, and show strong hydrophilicity as determined by thermodynamic analysis of the surface tension components. The contact of various challenging protein solutions results in repellency on the poly(DMLA) brushes. Furthermore, the poly(DMLA) brushes completely prevent the adhesion and colonization of Escherichia coli. Remarkably, upon blood contact, the poly(DMLA) brushes successfully prevent the formation of a fibrin network and leukocyte adhesion on the surface. While showcasing excellent antifouling properties similar to those of N-hydroxypropyl methacrylamide (HPMA) polymer brushes as one of the best antifouling coatings, the absence of hydroxyl groups prevents activation of the complement system in blood. We envision the polymer brushes to contribute to the future of hemocompatible coatings.

Blood Coagulation Activities of Cotton-Alginate-Copper Composites.

Alginate-based materials have gained significant attention in the medical industry due to their biochemical properties. In this article, we aimed to synthesize Cotton-Alginate-Copper Composite Materials (COT-Alg(-)Cu(2+)). The main purpose of this study was to assess the biochemical properties of new composites in the area of blood plasma coagulation processes, including activated partial thromboplastin time (aPTT), prothrombin time (PT), and thrombin time (TT). This study also involved in vitro antimicrobial activity evaluation of materials against representative colonies of Gram-positive and Gram-negative bacteria and antifungal susceptibility tests. The materials were prepared by immersing cotton fibers in an aqueous solution of sodium alginate, followed by ionic cross-linking of alginate chains within the fibers with Cu(II) ions to yield antimicrobial activity. The results showed that the obtained cotton-alginate-copper composites were promising materials to be used in biomedical applications, e.g., wound dressing.

Leukemia Inhibitory Factor, Tissue Factor, Thrombomodulin, P-Selectine, D-Dimer, Thrombin Generation and Procoagulant Phospholipid Dependent Clotting Time Are Predictive for in Vitro Fertilization Outcome. the Prospective Roadmap-IVF Study

Background The link between blood hypercoagulability, endothelial cell activation and infertility or in vitro fertilization (IVF) failure is a puzzling issue. Activation of platelets, endothelial cells and blood coagulation, proinflammatory and angiogenetic potential play important role in implantation and early embryonic development. Biomarkers of cellular hypercoagulability are associated with high risk of IVF failure. Soluble Leukemia Inhibitory Factor (LIF), an interleukin 6 class cytokine expressed in the trophectoderm of the developing embryo is an important regulator in the establishment of pregnancy. Endoglin (Eng), a co-receptor for TGF-β1 and TGF-β3 is implicated in vascular complications of pregnancy and particularly in preeclampsia. Aim: The prospective longitudinal monocentric observational cohort study ROADMAP-IVF, evaluated the synergistic prognostic value of LIF and Eng with the biomarkers of cellular hypercoagulability in predicting the endometrial receptivity of fresh IVF cycles. Materials and Methods. The ROADMAP-IVF enrolled 40 women eligible for IVF with normal blood count, PT, aPTT, Fg, renal and liver function. The control group (CG) consisted of 30 healthy women with history of uncomplicated pregnancies. Exclusion criteria: Use of anticoagulant or antiplatelet agents during the last 30 days before inclusion. Known cardiovascular disease, active cancer. Active corticosteroid treatment. Primary end-point: Echographically documented pregnancy at 7 weeks from implantation. Blood was collected on day 2 (D2) from the natural cycle and 6 days after hormone treatment initiation corresponding to D8 of the cycle. Procoagulant phospholipid-dependent clotting time (PPL-ct), tissue factor (TF), thrombomodulin (TM), von Willebrand factor (vWF) and D-Dimers (DDi) were measured with assays from Diagnostica Stago (Asnieres, France). P- and E-Selectin, LIF and Eng were measured with ELISA. Thrombin generation (TG) with the TF 5 pM PPP-Reagent ® was assessed on Calibrated Automated Thrombogram (Stago, France). The cut-off values for studied biomarkers have been selected on the basis of ROC analysis. Univariate and multivariate logistic regression analysis examined the associations between the biomarkers and the study outcome. Results: 40 women were enrolled in the study. In 11 women pregnancy was confirmed with ultrasound and in 23 women no pregnancy was echographically documented. Age was not significantly different between the IVF (41 ys; range 22 - 48 ys) and the control group (40 ys; range 20 - 46 ys). At D2 and D8 the DDi and LIF were significantly higher in IVFG as compared to the CG. At D2 most of the patients had PPL-ct, DDi and MRI of TG higher than the upper normal limit (UNL). At D8 only TF was significantly increased as compared to the CG. Analytical data are presented in Table 1. Age did not correlate with the levels of the studied biomarkers. Significant correlations (p<0.05) were found between DDi and ETP (r=0.35); TM, TF and P-Selectin (r=0.4) at D2; TF, LIF and Eng (r=0.5) at D2 and D8; P-Selectin and PPL-ct (r=-0.4) at D2; P-Selectin and TM (r=0.4), E-Selectin (r=0.5), LIF (0.4) at D2; E-Selectin and P-Selectin (r=0.5), TF (r=0.3), and LIF (r=0.4) at D2. The ROC analysis showed that IVF failure was associate with DDi increase at D2 (AUC = 0.64); P-Selectin increase at D2(AUC = 0.57) and D8 (AUC = 0.61); TM increase at D8 (AUC = 0.61); TF increase at D8 (AUC = 0.57); LIF increase at D8 (AUC = 0.59); ETP increase at D2 (AUC = 0.67) and D8 (AUC = 0.67); Peak increase at D2 (AUC = 0.64) and D8 (AUC = 0.61). Prolongation of PPL-ct on D8 was predictive for positive pregnancy outcome(AUC 0.64). Conclusion The ROADMAP-IVF study showed that at least one out of four women eligible for IVF present biological evidence of activation of platelets, endothelial cells, blood coagulation or LIF upregulation. Hormone treatment administration does not significantly alter the proinflammatory or the hypercoagulable state. Among the studied biomarkers the levels of LIF, DDi, P-Selectine, TM and TF as well as the status of thrombin generation and PPL-ct showed a significant predictive value for the IVF outcome, particularly when measured at 6 days after hormone treatment administration. These findings will lead to the elaboration of an risk assessment model for IVF failure combining the selected biomarkers of hypercoagulability with LIF.

A Syngeneic Murine Model of Antiphospholipid Syndrome (APS) Recapitulates Aspects of Human APS

Background Antiphospholipid syndrome (APS) is characterized by arterial or venous thrombosis, or recurrent pregnancy loss, in the presence of persistently positive tests for antiphospholipid antibodies (aPL). aPL include lupus anticoagulants (LAC), anti-β2-glycoprotein 1 (β2GPI) and anti-cardiolipin antibodies (aCL). Among them, antibodies directed toward β2GPI have been best characterized and shown in most studies to have a significant association with thrombosis. Most murine models of APS are generated by infusion of either polyclonal IgG isolated from APS patients, or monoclonal IgG directed against human or murine β2GPI. However, one or more of the following limitations may limit their clinical relevance: 1) Unlike chronically elevated autologous aPL in APS patients, cross-species IgG infused into mice circulate for only limited periods of time and may provoke allogeneic responses; 2) Monoclonal human or murine anti-β2GPI IgG, unlike polyclonal aPL from patients, tend to be high affinity, monospecific, and may recognize epitopes different than those recognized by clinical aPL. Hence, we sought to develop a relevant syngeneic murine model of APS that employed polyclonal murine anti-β2GPI IgG that would circulate in mice for extended periods, and not induce allogeneic inflammatory responses. Methods β2GPI-deficient ( apoh -/-) mice were previously developed in our lab on a C57/BL6 background using CRISPR/cas9. IgG isolated from apoh -/- mice immunized with human plasma β2GPI was used as a source of polyclonal murine aPL IgG, while IgG isolated from apoh -/- mice treated with adjuvant alone served as control IgG in all experiments. Wild-type (WT) C57/BL6 mice (two groups of 8 mice each) were injected intravenously with 20 µg of either murine polyclonal aPL or control IgG, and after 72 hours were evaluated for venous thrombosis using the IVC stasis model, with thrombi harvested and measured 48 h after IVC ligation. In parallel studies, we injected mice with aPL or control IgG and evaluated 1) activation of washed platelets, 2) aortic wall gene expression, and 3) levels of circulating neutrophil extracellular traps (NETs) using blood and tissue samples isolated at identical time points as IVC ligation studies (mice in these studies did not undergo IVC ligation). Platelet activation was assessed by measuring the binding of JON/A and expression of platelet P-selectin by flow-cytometry. Total RNA isolated from aortic walls of 3 mice each treated with murine aPL or control IgG was analyzed by RNA sequencing. Results WT C57/BL6 mice that received 20 µg of polyclonal murine aPL sustained plasma anti-β2GPI IgG levels >500 ng/mL for more than 120 h after IgG injection. In the IVC stasis model, six out of eight mice receiving aPL IgG and four out of eight mice receiving control IgG developed thrombi (lack of thrombi in all mice was likely due to bleeding of extensive IVC back branches in C57/BL6 mice). Thrombi weight in aPL-treated mice were significantly larger than those from mice that received control IgG (19.7 ± 1.7 mg vs 13.5 ± 1.0 mg, P = 0.015). Thrombin (0.1 U/ml)-induced platelet activation was significantly greater for platelets isolated from aPL-administered mice compared to those from control mice 72h after IgG injection as determined by binding of JON/A (1246.7 ± 55.3 MFI vs 879 ± 37.9 MFI, P = 0.005) and expression of P-selectin (5729.7 ± 553.9 MFI vs 3777.3 ± 402.5 MFI, P=0.046). Gene enrichment analysis of mRNA from aortas demonstrated significant upregulation of gene pathways related to complement and coagulation cascades, blood coagulation, and oxidoreductase activity in the aPL-treated mice. Individually upregulated genes included C8, C9 fga and fgb. Elevated levels of circulating nucleosomes were also observed in aPL treated mice but did not reach significance due to small numbers. Conclusion These results demonstrate that anti-β2GPI polyclonal aPL raised in apoh-/- mice circulate for extended periods when reinfused into WT mice, enhance thrombosis, induce platelet hyperreactivity, and cause changes in mRNA expression in the vascular wall. Further characterization of anti-β2GPI IgG for LAC activity, anti-PS/PT reactivity and β2GPI domain 1 reactivity is in progress. While all animal models of APS have weaknesses, this syngeneic model provides an additional approach that recapitulates many aspects of vascular inflammation seen in human APS.

Sterilized Polyhexanide-Releasing Chitosan Membranes with Potential for Use in Antimicrobial Wound Dressings.

Wound infection is a common complication of chronic wounds. It can impair healing, which may not occur without external help. Antimicrobial dressings (AMDs) are a type of external help to infected chronic wounds. In this study, highly porous membranes made of only chitosan and containing the antiseptic polyhexanide (poly(hexamethylene biguanide); PHMB) were prepared by cryogelation, aiming to be used in AMDs. These membranes exhibited a water swelling capacity of 748%, a water drop penetration time of 11 s in a dry membrane and a water vapor transmission rate of 34,400 g H2O/m2/24 h when in contact with water. The best drug loading method involved simultaneous loading by soaking in a PHMB solution and sterilization by autoclaving, resulting in sterilized, drug-loaded membranes. When these membranes and a commercial PHMB-releasing AMD were assayed under the same conditions, albeit far from the in vivo conditions, their drug release kinetics were comparable, releasing PHMB for ca. 6 and 4 h, respectively. These membranes exhibited high antibacterial activity against Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa, which are bacterial species commonly found in infected wounds and blood clotting activity. The obtained results suggest that these membranes may have potential for use in the development of AMDs.

GRSF1 antagonizes age-associated hypercoagulability via modulation of fibrinogen mRNA stability

Age-associated hypercoagulability is accompanied by the increase of plasma levels of some coagulation factors including fibrinogen which may contribute to the increased risk of cardiovascular, cerebrovascular, and thrombotic diseases in elderly people. However, the underlying mechanism of increased plasma fibrinogen concentration during aging is still elusive. GRSF1 belongs to the heterogeneous nuclear ribonucleoproteins F/H (hnRNP F/H) subfamily. Here, we report that GRSF1 attenuates hypercoagulability via negative modulation of fibrinogen expression. We demonstrated that GRSF1 negatively regulated fibrinogen expression at both mRNA and protein levels. GRSF1 directly interacted with the coding region (CDS) of FGA, FGB, and FGG mRNAs, and decreased their stability thus mitigating fibrinogen expression. We further identified that only a few G-tracts within the Fib C domain of FGA, FGB, and FGG CDS and the qRRM2 domain of GRSF1 were required for their interaction. Moreover, we confirmed hypercoagulability and the decrease of GRSF1 expression level during mice aging. Functionally, GRSF1 overexpression in old mice liver decreased fibrinogen plasma level, reduced hypercoagulability, and mitigated blood coagulation activity, whereas GRSF1 knockdown in young mice liver increased fibrinogen plasma level and promoted blood coagulation activity. Collectively, our findings unveil a novel posttranscriptional regulation of fibrinogen by GRSF1 and uncover a critical role of GRSF1 in regulating blood coagulation activity.

Activation of intravascular blood coagulation in the early postoperative period in reconstructive abdominoplasty

Purpose of the study: to determine the activity of intravascular blood coagulation during the abdominoplasty and justify pharmacological thromboprophylaxis.Material and Methods. An open prospective study was conducted on a group of 100 patients undergoing abdominoplasty. The risk of venous thromboembolism (VTE) before the surgery was assessed using the Caprini risk assessment model in 2010. Prothrombin (PT) and fibrinogen (FG) levels, soluble fibrin-monomer complexes (SFMC), euglobulin clot lysis time (ECLT), antithrombin III (AT-III) level, and protein C (PC) level were determined before and after the surgery.Results. Using the Caprini risk assessment model, patients were categorized by VTE risk as follows: moderate risk – 64% and high risk – 36%. After the surgery, an increase in PT by 12.3% and 15.5% on days 3 and 5 was  observed (p < 0.01); an increase in FG by 0.49; 2.34 and 2.04 g/L on days 1, 3, and 5 was observed (p < 0.01); an increase in SFMC at all postoperative stages by 4.7; 8.2; 10.8 and 10.0 times (p < 0.01); an increase in ECLT by 2.8; 9.7, and 6.3 minutes on days 1, 3, and 5 (p < 0.001); and a decrease in AT-III and PC levels immediately after  the operation by 18.8% and 8.9%, on day 1 by 14.2% and 11.2%, and on day 3 by 8.8% and 6.3% (p < 0.01).Conclusion. All patients undergoing abdominoplasty require pharmacological thromboprophylaxis due to moderate and high risk of venous thromboembolic complications. Abdominoplasty in the early postoperative period is accompanied by activation of plasma coagulation, inhibition of fibrinolytic system activity, and reduced activity of natural anticoagulants, which justifies the need for pharmacological thromboprophylaxis for at least 5 days.

In vivo genome editing using 244-cis LNPs and low-dose AAV achieves therapeutic threshold in hemophilia A mice

Gene therapy and rebalancing therapy have emerged as promising approaches for treating hemophilia A, but there are limitations, such as temporary efficacy due to individual differences. Genome editing for hemophilia has shown long-term therapeutic potential in preclinical trials. However, a cautious approach is necessary because genome editing is irreversible. Therefore, we attempted to induce low-level human factor 8 (hF8) gene knockin (KI) using 244-cis lipid nanoparticles and low-dose adeno-associated virus to minimize side effects and achieve a therapeutic threshold in hemophilia A mice. We selected the serpin family C member 1, SerpinC1, locus as a target to enable a combined rebalancing strategy with hF8 KI to augment efficacy. This strategy improved blood coagulation activity and reduced hemophilic complications without adverse effects. Furthermore, hemophilic mice with genome editing exhibit enhanced survival for 40weeks. Here, we demonstrate an effective, safe, and sustainable treatment for hemophilia A. This study provides valuable information to establish safe and long-term genome-editing-mediated treatment strategies for treating hemophilia and other protein-deficient genetic diseases.

PB0791 Bacterial Biofilms Produced by Staphylococcus sp. Act as Molecular Scaffolds for the Activation of Blood Coagulation

PB0791 Bacterial Biofilms Produced by Staphylococcus sp. Act as Molecular Scaffolds for the Activation of Blood Coagulation

Characteristics of FXa-storing platelets in hemophilia B mice and the influence of alcohol on the platelets

Platelet-stored activated blood coagulation factor X (FXa) has great potential in the gene therapy of hemophilia B (HB). However, we still need to understand more about the properties of FXa-storing platelets and how dietary factors affect them. We created transgenic mice called 2bFXa-HB, which had stable expression and storage of FXa in their platelets, resulting in the alleviation of the bleeding disorder in these mice. Even after inducing anti-factor IX (FIX) inhibitors in 2bFXa-HB mice, the hemorrhage phenotype could still be rescued by the expression of FXa. The activation capacity of 2bFXa-HB platelets remained unchanged, and there were no signs of elevated thrombotic risk in these mice. In an acute alcohol exposure mouse model, a single administration of alcohol reduced both the number of platelets and their activation capacity, as well as impaired coagulation function. However, it did not increase the markers of thrombotic risk in either 2bFXa-HB or HB mice. These results suggest that FXa storage in platelets is safe and effective for treatment of HB, but alcohol could impair the therapeutic effect of FXa-containing platelets.

Exploration of Anthelmintic, Blood Coagulant, Diuretic and Laxative Activities of Different Solvent Fractions of Flagellaria Indica Leaves

Flagellaria indica (Family: Flagellariaceae) is a common climbing plant found primarily in tropical regions of many countries. The plant has various traditional uses, although most of them lack scientific published reports. The crude ethanolic extract of F. indica leaves was fractionated based on polarity using water, ethyl acetate, and n-hexane. Biological screening was conducted on the anthelmintic, blood coagulation, diuretic, and laxative activities of the water, ethyl acetate, and n-hexane fractions of F. indica leaves. In the anthelmintic test, the n-hexane fraction showed a moderate effect with paralysis times of 16.79 and 13.62 minutes and death times of 27.34 and 21.81 minutes, respectively, at doses of 25 and 50 mg/mL.In the blood coagulant test, only the water fraction showed a notable effect. The clotting times were 4.33, 6.02, 7.68, and 8.32 minutes, respectively, at doses of 200, 100, 50, and 25 mg/mL. Diuretic activity was performed to determine the increase in the volume of excreted urine, and electrolyte analysis of urine was performed to determine pH, density, conductance, and Na+, Cl-, K+ levels, as well as natriuretic, kaliuretic, saluretic, and CAI indexes. The ethyl acetate fraction showed better diuretic activity than the n-hexane fraction, while the water fraction did not reveal a notable diuretic effect. The Na+, Cl-, K+, natriuretic, and saluretic indexes were found satisfactory in the ethyl acetate fraction, and the CAI index was better in the n-hexane fraction. In the laxative test, the n-hexane fraction showed the best laxative properties, with an increase in stool weight of 38% and 54% at doses of 250 and 500 mg/kg, respectively. These results suggest that different fractions of F. indica leaves contain distinct phytochemicals that may be responsible for these biological effects. The isolation of bioactive compounds could help justify its traditional uses in modern medicine.

Platelets provide robustness of spatial blood coagulation to the variation of initial conditions

Activated platelets provide phospholipid surface and secrete coagulation factors, enhancing blood clotting. We investigated the role of platelets in the regulation of blood coagulation spatial dynamics. We activated blood clotting with tissue factor-bearing (TF) surface in platelet-rich plasma (PRP) or platelet-free plasma (PFP). When blood coagulation was initiated by high TF density, clot growth rate (V) in PRP (2 × 105/μL platelets) was only 15 % greater than in PFP. Spatial distribution of thrombin in PRP had a peak-like shape in the area of the fibrin clot edge, while in PFP thrombin was distributed in the shape of descending plateau. Platelet inhibition with prostaglandin E1 or cytochalasin D made spatial thrombin distribution look like in the case of PFP. Inhibition of blood coagulation by natural endogenous inhibitor heparin was diminished in PRP, while the effect of the exogenous or artificial inhibitors (rivaroxaban, nitrophorin, hirudin) remained undisturbed in the presence of platelets. Ten times decrease of the TF surface density greatly depressed blood coagulation in PFP. In PRP only clotting initiation phase was, while the propagation phase remained intact. Coagulation factor deficiency greatly reduced amount of thrombin and decreased V in PFP rather than in PPR. Thus, platelets were redundant for clotting in normal plasma under physiological conditions but provided robustness of the coagulation system to the changes in initial conditions.

Immunological, hormonal and hemostasis parameters in cosmonauts’ blood after long-term orbital missions

The extreme factors associated with space flight influence pronouncedly various physiological systems of the human body. The current study focuses on the impact of factors associated with prolonged space missions lasting up to 6 months on cellular innate immunity, hormonalregulation, and hemostasis in Russian cosmonauts. Immune system adaptation to the conditions of long-term space flight was shown to be accompanied by a significant increase in the level of monocytes and granulocytes, along with a decreased number of cells expressing adhesion molecules. The coagulation cascade, main anticoagulant and fibrinolysis proteins were also evaluated. The early post-flight period was characterized by elevated D-dimer levels compared to baseline values, indicative of blood clotting and fibrinolysis activation caused by a damage of endothelium during landing. aAlterations in blood rheology during re-adaptation to Earth conditions was also observed. The blood concentrations of aldosterone, cortisol, estradiol, testosterone, osteocalcin, and erythropoietin were analyzed, and their correlation with the parameters of hemostasis was assessed.The summary findings indicate an adaptive physiological reaction in cosmonauts associated with 6 months duration of spaceflight. The observations may be related to endothelium damage, which manifested in the activation of certain components of the immune system, along with the activation of fibrin clot formation and subsequent fibrinolysis.

Features of disorders and methods of correction of the hemostasis system in women with antenatal fetal death

The improvement of the providing medical services quality to pregnant women remains an urgent issue in modern obstetrics. Women with perinatal losses deserve special attention, especially in the second half of pregnancy. Changes in the hemostasis system are an integral part of the development of pregnancy. The tendency to hypercoagulation has significant pathogenetic significance and can be the cause of a number of complications – miscarriage in the I trimester of pregnancy and preeclampsia, premature birth, antenatal fetal death (AFD) – in the II and III trimesters of pregnancy. Also, pregnancy is a background process for the activation of a number of diseases, in particular hereditary thrombophilia.The objective: to evaluate the changes in the hemostasis system in women with antenatal fetal death and the effectiveness of the use of low molecular weight heparins for the correction of disorders in the hemostasis system in this category of patients during childbirth and in the postpartum period.Materials and methods. 72 women were examined, including 42 pregnant women with AFD (main group) and 30 pregnant women with a physiological course of pregnancy, childbirth and the postpartum period (control group). The state of the hemostasis system was studied using a standard coagulogram. D-dimer was determined by immunoturbometric analysis.To assess the state of the hemostasis system, the following biochemical tests were used: procoagulant link – fibrinogen content and indicators: prothrombin index, activated partial thrombin time (APTT), thrombin time (TP), ancistrone time (AT), soluble fibrinogen-monomer complex (SFMC), factor X (FX); to evaluate the antithrombin system, the content of antithrombin-III (AT-III), protein C were determined; to characterize the state of the fibrinolytic system – the amount of plasminogen, α2-antiplasmin, fibrinogen degradation products (FDC).Thrombophilia markers and antiphospholipid antibodies were also determined. The preference was given to vaginal childbirth. During childbirth, mechanical compression of the lower limbs was applied using special compression stockings (compression level 2). In 12 hours after delivery thromboprophylaxis with low molecular weight heparins was started, the drug of choice being enoxaparin sodium. After 48 hours of the postpartum period, a comparative analysis of coagulogram indicators was performed to further determine the timing of thromboprophylaxis.Results. Analysis of family thrombotic anamnesis revealed risk factors in 12 (28.6%) patients of the main group. In close relatives of the first line, the presence of peripheral vein thrombosis was found – 5 (41.7%) cases, myocardial infarction under the age of 45 – 3 (25.0%) cases, pulmonary embolism – 1 (8.3%) case, transient ischemic attack of the brain – 3 (25.0%).In the control group only 2 (6.6%) patients had a family history of venous thromboembolism in first-line relatives. The analysis of the coagulation system shows significantly higher fibrinogen values (the main group – 5.3±0.2 g/l, the control group – 4.3±0.1 g/l; p<0.05), functional FX activity (the main group – 149.3±3.1%, control group – 107.3±2.7%; p<0.05), SFMC (main group – 15.9±1.2 μg/ml, control group – 7.8±0.9 μg/ml; p<0.05) and D-dimer (main group – 4.4±0.25 μg/ml, control group – 0.7±0.2 μg/ml; p< 0.05) in pregnant women of the main group. An increase in these indicators is a predictor of thrombus formation, activation of blood coagulation by the internal pathway with a decrease in the antithrombin reserve due to FX.During the correlation analysis, a strong direct relationship (r=0.8633) was established between the indicators of SFMC and FX in the blood serum of pregnant women of the main group, the combination of which determines the tendency to clot formation. Determination of markers for the most common types of thrombophilia shows the dominance of hereditary forms (prothrombin, Leiden mutation, MTHFR) in 43% of pregnant women of the main group.The implementation of the proposed treatment approach contributed to a significant decrease in the average concentration of fibrinogen 48 hours after delivery in postpartum women of the main group (main group: before delivery – 5.3±0.2 g/l, after 48 hours after delivery – 3.9±0.3 g/l; control group: 48 hours after delivery – 3.2±0.2 g/l; p<0.05) in combination with a synergistic decrease in the average indicators of SFMC (main group: before delivery – 15.9±1.2 μg/ml, 48 hours after delivery – 6.2±0.2 μg/ml; control group: 48 hours after delivery – 5.4±0.3 μg/ml; p<0.05) and FX (main group: before delivery – 149.3±3.1%, after 48 hours after delivery – 103.1±3.6%; control group: after 48 hours after delivery – 117.1±4.1%; p<0.05).Conclusions. Increased coagulation function was confirmed in pregnant women with antenatal fetal death (AFD). Thanks to the proposed method of correcting disorders in the hemostasis system, a decrease in the frequency of postpartum thromboembolism in women with AFD is achieved, as well as an improvement in the main indicators of the hemostasis system. This prevents the emergence of a chronic form of the syndrome of disseminated intravascular blood coagulation which is developed in the cases of AFD.

Effects of sesame (Sesamum indicum L.) and bioactive compounds (sesamin and sesamolin) on inflammation and atherosclerosis: A review.

Inflammation, oxidative stress, obesity, infection, hyperlipidemia, hypertension, and diabetes are the main causes of atherosclerosis, which in the long term lead to hardening of the arteries. In the current study, we reviewed recent findings of the mechanism of sesame and its active compounds of sesamin and sesamolin regulates on atherosclerosis. Sesame can decrease the lipid peroxidation and affect the enzymes, which control the balance of oxidative status in the body. Besides modulating the inflammatory cytokines, sesame regulates the main mediators of the signaling pathways in the process of inflammation, such as prostaglandin E2 (PGE2), nuclear factor kappa light-chain enhancer of activated B cells (NF-kB) and peroxisome proliferator-activated receptor gamma (PPAR-γ). Sesame decreases the growth of different pathogens. It fights against obesity and helps to reduce weight, body mass index (BMI), waist circumference, and lipid count of serum and liver. In addition to lowering fasting blood sugar (FBS), it decreases the hemoglobin A1c (HbA1c) and glucose levels and improves insulin function. With high content of linoleic acid, α-linolenic acid, and total polyunsaturated fatty acid (PUFA), sesame efficiently controls the blood plasma lipids and changes the lipid profile. In the case of hypertension, it maintains the health of endothelium through multiple mechanisms and conserves the response of the arteries to vasodilation. PUFA in sesame suppresses blood clotting and fibrinogen activity. All the mentioned properties combat atherosclerosis and hardening of blood vessels, which are detailed in the present review for sesame.