Search Results (80)

This study aimed to characterize and evaluate the fibrinolytic, thrombolytic, hematological, and toxicological aspects of a serine protease (AsKSP) from Aspergillus tamarii Kita UCP 1279. The enzyme was purified using a two-phase aqueous system and assessed for optimal pH (7.0) and temperature (50 °C), stability, and effects of metal ions, inhibitors, and surfactants. AsKSP exhibited stability for up to 120 min at 50 °C and 36 h at pH 7.0. Enzymatic activity was enhanced by Na⁺ and Zn²⁺ and non-ionic surfactants (Tween-80) but inhibited by Cu²⁺, Fe³⁺, Triton X-100, and SDS, reducing activity by up to 62.35%. The highest amidolytic activity was observed for the substrate N-succinyl-Gly–Gly–Phe-p-nitroanilide. SDS-PAGE analysis indicated an approximate molecular mass of 90 kDa. The enzyme showed fibrinolytic activity, degrading 38.81% of fibrin clots in vitro after 90 min, without affecting fibrinogen. Cytotoxicity assays indicated no toxicity (cell viability > 80%). Coagulation assays showed slight prolongation of prothrombin time (PT) and activated partial thromboplastin time (aPTT), with no effect on thrombin time. No red blood cell lysis was observed, and albumin increased enzymatic activity by 31.70%. These findings demonstrate that Aspergillus tamarii Kita UCP 1279 produces a fibrinolytic protease with potential for thrombus treatment, providing a promising foundation for drug development. Full article

Dengue virus is the most prevalent arthropod-borne viral disease in humans. Severe dengue, defined by hemorrhagic fever and dengue shock syndrome, can develop quickly in people who have warning indications such as abdominal pain, mucosal bleeding, and a significant decrease in platelet count. Laboratory markers such as hematocrit, platelet count, liver enzymes, and coagulation tests are critical for early diagnosis and prognosis. This retrospective study was carried out from January 2023 to December 2024 at a super-specialty tertiary care hospital. There were 283 adult patients with dengue with warning signs, who were categorized into 102 with platelet transfusion and 181 with no platelet transfusion. Data on patient demographics, clinical history, laboratory values, and radiological findings were systematically obtained from hospital records at the time of admission. Laboratory parameters such as white blood cell (OR = 2.137), hemoglobin (OR = 2.15), aPTT (OR = 5.815), AST²/ALT (OR = 2.431), platelet count (OR = 26.261) and NS1 (OR = 4.279) were found to be significantly associated (p < 0.01) with platelet transfusion. Similarly, an increased prothrombin time (OR = 2.432) contributed to prolonged hospital stays and the presence of ascites (OR = 5.059), gallbladder wall thickening (OR = 4.212), and pleural effusion (OR = 2.917), contributing to the severity of the dengue infection. These significant laboratory markers help with identifying patients with dengue who may develop severe dengue, requiring platelet transfusion, thereby prioritizing patient care and enabling the implementation of targeted interventions. Full article

A 63-year-old woman with rheumatoid arthritis and Hashimoto’s thyroiditis was admitted to the emergency room, because of left leg pain associated with spontaneous subcutaneous hematomas, for 15 days. Their symptoms also occurred after the discontinuation of aspirin, which the patient had taken for a previous case of ocular papillitis. Laboratory tests showed anemia, a normal platelet count, but a prolonged activated partial thromboplastin time (aPTT) ratio; a computerized tomography scan of the left lower limb detected a recent hematoma in the left lateral rectus muscle, and subcutaneous soft tissue edema also involving the knee, without vascular involvement. Coagulation tests were performed showing normal levels of Lupus Anticoagulant, very low-factor FVIII activity (2.2%), normal FIX, FXI, and FXII activity, and the detection of FVIII inhibitors by a Bethesda assay (7.6 U). A diagnosis of acquired hemophilia A (AHA) was made, and hemostatic and immunosuppressive treatment was immediately started (activated prothrombin complex concentrates and methylprednisolone). Malignancies and infections were excluded. An autoantibodies panel confirmed the positivity to rheumatoid factor and anti-cyclic citrullinated peptide antibodies. In treatment, the patient did not present any new bruises, with aPTT normalizing, FVIII increasing, and inhibitors reducing until disappearance. A close follow-up continued every 1–2 week after discharge, with hemostatic treatment discontinuation and methylprednisolone decalage. Underlying autoimmune conditions induced this rare, autoimmune and life-threating disorder. Full article

Cardiovascular diseases, including thrombosis, are the leading cause of mortality worldwide. The generation of monoclonal antibodies (mAb) targeting specific coagulation factors could provide more targeted and safer anticoagulant therapies. Factor V (FV) is a critical cofactor in the prothrombinase complex, which catalyzes the conversion of prothrombin to thrombin, a key enzyme in the coagulation cascade. We isolated a novel human antibody specific to FV by using phage display technology. The selection occurred by panning a large repertoire of phages expressing human antibody fragments (scFv) in parallel on the purified recombinant protein in its native form (FV) or activated by proteolytic maturation (Factor Va (FVa)). Through ELISA screening, we identified the clone with the highest binding affinity for both targets, and it was successfully converted into IgG1. The novel human mAb, called D9, was found capable of binding to Factor V with a low nM affinity both by ELISA and BLI assays, whereas its cross-reactivity with some other coagulation factors was found null or very poor. Furthermore, when tested in blood clotting tests, it was found able to prolong activated partial thromboplastin time (aPTT). Thus, D9 could become not only a potential therapeutic agent as a specific anticoagulant but also a precious tool for diagnostic and research applications. Full article

The primary challenges in the tissue engineering of small-diameter artificial blood vessels include inadequate mechanical properties and insufficient anticoagulation capabilities. To address these challenges, urea-pyrimidone (Upy)-based polyurethane elastomers (PIIU-B) were synthesized by incorporating quadruple hydrogen bonding within the polymer backbone. The synthesis process employed poly(L-lactide-ε-caprolactone) (PLCL) as the soft segment, while di-(isophorone diisocyanate)-Ureido pyrimidinone (IUI) and isophorone diisocyanate (IPDI) were utilized as the hard segment. The resulting PIIU-B small-diameter artificial blood vessel with a diameter of 4 mm was fabricated using the electrospinning technique, achieving an optimized IUI/IPDI composition ratio of 1:1. Enhanced by multiple hydrogen bonds, the vessels exhibited a robust elastic modulus of 12.45 MPa, an extracellular matrix (ECM)-mimetic nanofiber morphology, and a high porosity of 41.31%. Subsequently, the PIIU-B vessel underwent dual-functionalization with low-molecular-weight heparin and gelatin via ultraviolet (UV) crosslinking (designated as PIIU-B@LHep/Gel), which conferred superior biocompatibility and exceptional anticoagulation properties. The study revealed improved anti-platelet adhesion characteristics as well as a prolonged activated partial thromboplastin time (APTT) of 157.2 s and thrombin time (TT) of 64.2 s in vitro. Following a seven-day subcutaneous implantation, the PIIU-B@LHep/Gel vessel exhibited excellent biocompatibility, evidenced by complete integration with the surrounding peri-implant tissue, significant cell infiltration, and collagen formation in vivo. Consequently, polyurethane-based artificial blood vessels, reinforced by multiple hydrogen bonds and dual-functionalized with heparin and gelatin, present as promising candidates for vascular tissue engineering. Full article

Background: The most important concern regarding living donor liver transplantation is the safety of living liver donors, of which anesthesia management is an important part. Sugammadex, which has recently been used frequently for the reversal of neuromuscular blockade, is known to cause adverse effects on the coagulation profile. This study seeks to assess the impact of neostigmine and sugammadex on coagulation parameters in living liver donors following hepatectomy. Methods: We compared the demographic, clinical, and coagulation parameters of 209 living liver donors who received sugammadex (2 mg/kg) for neuromuscular blockade reversal during donor hepatectomy procedures from January 2018 to July 2022, with 209 patients who received neostigmine (50 g/kg) for the same purpose during the same timeframe. We compared the following parameters: age, gender, prothrombin time (PT), partial thromboplastin time (PTT), international normalized ratio (INR), hemoglobin (Hb), platelet count, ICU stay, hospital stay, and relaparotomy for bleeding and other causes. Results: Demographic data and preoperative biochemical values were similar in both groups. PT (p = 0.004) and aPTT (p < 0.001) values were significantly longer in the postoperative period in both groups; the difference between preoperative and postoperative PT (p = 0.009) and aPTT (p < 0.001) was significantly higher in the sugammadex group. However, neither group showed any elongation beyond the reference range. The sugammadex group had an elevated postoperative platelet count (p = 0.040). The duration of patients’ stay in the ICU was significantly shorter in the sugammadex group (p < 0.001). Conclusion: The prolonged aPTT and PT associated with sugammadex did not lead to any postoperative bleeding complications. The sugammadex group significantly reduced the duration of ICU stays, while the hospital stays remained comparable. Further multicentric prospective randomized studies should support our study’s findings, which demonstrate the safe use of low-dose sugammadex. Full article

Xylans, polysaccharides abundantly derived from agricultural byproducts, have shown potential pharmacological properties, making them a subject of increasing research interest. This study aimed to expand the understanding of xylans’ pharmacological properties and relate them to their composition. A method combining ultrasound and alkaline media for xylan extraction from corn cobs (ERX) was used, resulting in a significant increase in final yield compared to other methodologies. The physicochemical characterization of ERX was carried out, and its antioxidant, cytotoxic, anticoagulant, and immunomodulatory properties were evaluated. ERX demonstrated significant antioxidant activity with metal-chelating properties and induced apoptosis in HeLa tumor cells (p < 0.0001). It also reduced nitric oxide (NO) production by activated macrophages and extended the blood coagulation time, as assessed by the APTT assay (p < 0.0001). Further fractionation of ERX using various organic solvents resulted in multiple xylan subfractions. Among them, the ethanol-derived subfraction E1.4 exhibited remarkable pharmacological activities, including metal-chelation, cytotoxicity against HeLa cells via apoptosis, reduced NO production (p < 0.0001), and prolonged coagulation times (p < 0.0001). E1.4 is heteroxylan with a molecular weight of approximately 100 kDa. These findings suggest that corn cobs could be a promising source of pharmacologically significant molecules, particularly the heteroxylan E1.4. Future studies should focus on the structural characterization of this xylan to understand the relationship between structure and biological activity and explore the therapeutic potential of E1.4 in vivo models. Full article

Granule secretion is an essential platelet function that contributes not only to haemostasis but also to wound healing, inflammation, and atherosclerosis. Granule secretion from platelets is facilitated, at least in part, by Soluble N-ethylmaleimide-Sensitive Factor (NSF) Attachment Protein Receptor (SNARE) complex-mediated granule fusion. Although α-synuclein is a protein known to modulate the assembly of the SNARE complex in other cells, its role in platelet function remains poorly understood. In this study, we provide evidence that α-synuclein is critical for haemostasis using α-synuclein-deficient (^−/−) mice. The genetic deletion of α-synuclein resulted in impaired platelet aggregation, secretion, and adhesion in vitro. In vivo haemostasis models showed that α-synuclein^−/− mice had prolonged bleeding times and activated partial thromboplastin times (aPTTs). Mechanistically, platelet activation induced α-synuclein serine (ser) 129 phosphorylation and re-localisation to the platelet membrane, accompanied by an increased association with VAMP 8, syntaxin 4, and syntaxin 11. This phosphorylation was calcium (Ca²⁺)- and RhoA/ROCK-dependent and was inhibited by prostacyclin (PGI₂). Our data suggest that α-synuclein regulates platelet secretion by facilitating SNARE complex formation. Full article

Few studies are concerned with the effect of the conjugat protein on the bioactivities of the abalone gonad polysaccharide (AGP). In this study, a series of treatments, including raw material (female and male) defatting, extraction temperature (25–121 °C), proteolysis, ultrafiltration, and ethanol precipitation, was conducted to investigate the role of the conjugate protein on AGP anticoagulant activity. All AGP extracts significantly prolonged activated partial thromboplastin time (APTT) and thrombin time (TT). The strongest was observed in the female AGPs prepared at 50 and 121 °C. The most active is located at 30–300 kDa by ultrafiltration. After being exposed to neutral protease, quick shortening of APTT and TT was found in all AGPs. Further ethanol precipitating of found the longest APTT in the sediment, which contains most polysaccharides and proteins. Defatting lowered the activity of female AGP but increased that of males. Proteolysis also significantly weakened the clotting factor inhibition effect of the 50 °C female AGP, but heating seemed not affect the effect. Five fractions were obtained after the 50 °C female AGP was subjected to ion exchange column. Fraction V, with the highest protein and medium polysaccharide content, showed the strongest anticoagulant effect and was also much higher than AGSP, which was obtained by multi-step proteolysis. The findings supported positive effect of the conjugate protein in AGP anticoagulant activity. Full article

This study introduces a novel approach for enhancing the functional properties of cotton fibers through complexation of copper sulfate, and subsequent combination with chitosan (COT-CuSO₄-CTS). Our preliminary investigations focused on the development composites as candidate materials for functional coatings with antimicrobial properties. The materials were thoroughly characterized via scanning electron microscopy (SEM) and optical microscopy, providing insights into their structural features and composition. The findings show that the modified cotton materials exhibit potent antimicrobial activity. Specifically, the COT-CuSO₄ and COT-CuSO₄-CTS samples demonstrated zones of inhibition against both Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli, confirming their ability to reduce microbial growth significantly. The incorporation of a chitosan layer significantly enhanced the Ultraviolet Protection Factor (UPF) of the cotton fabric from 3.37 to over 50, indicating exceptional UV shielding capabilities, while copper(II) oxide treatment provided a moderate UPF value of 14.56. Blood compatibility studies further revealed that COT-CuSO₄ and COT-CuSO₄-CTS fabrics influence coagulation parameters, with a marked prolongation in activated partial thromboplastin time (aPTT) and prothrombin time (PT) compared to untreated cotton. This anticoagulant effect is primarily linked to the presence of copper, although the addition of chitosan modulates this response, slightly reducing clotting times compared to COT-CuSO₄ alone. Cytotoxicity and genotoxicity assessments using Peripheral Blood Mononuclear (PBM) cells indicated that untreated cotton was non-toxic and non-genotoxic. However, COT-CuSO₄ and COT-CuSO₄-CTS fabrics displayed a reduction in cell viability and induced DNA damage, highlighting their potential cytotoxic and genotoxic effects. Notably, COT-CuSO₄-CTS showed lower cytotoxicity and genotoxicity than COT-CuSO₄-CTS, suggesting that chitosan reduces the overall cytotoxic and genotoxic potential of the composite. Furthermore, plasmid DNA relaxation assays indicated that COT-CuSO₄ and COT-CuSO₄-CTS interact with DNA, with COT-CuSO₄ exhibiting a stronger interaction than COT-CuSO₄-CTS, consistent with the findings on PBM cells. Full article

Phyllophorus proteus is a low-value sea cucumber from Indonesia and other tropical peripheral waters. In this study, a fucosylated glycosaminoglycan (FG) was extracted from P. proteus. It consists of GlcA, GalNAc, and Fuc, with a molecular weight of 67.1 kDa. The degraded FG (dFG) was prepared by β-elimination. Structural analysis revealed that the main chain of dFG was composed of GalNAc and GlcA, linked alternately by β1,3 and β1,4 glycosidic bonds. The sulfate group was located at the 4 and 6 positions of GalNAc. Fuc was attached to the 3 position of GlcA by an α1,3 glycosidic bond, and the side chain of Fuc exhibited various sulfate substitutions. FG significantly prolonged the coagulation time of APTT, PT, TT, and FIB, surpassing the effect of LMWH, thereby demonstrating its ability to exert anticoagulant effects in both the endogenous and exogenous coagulation pathways. Conversely, dFG had no significant effect on the clotting time of PT, suggesting its lack of impact on the intrinsic coagulation pathway. This study elucidates the structural properties and potent anticoagulant activities of fucosylated glycosaminoglycan from P. proteus. Full article

Direct oral anticoagulants (DOACs) are increasingly used for the treatment of thrombosis. While inhibitors of factor IIa and factor Xa have shown effectiveness, the risk of bleeding remains a significant concern. Recently, direct factor XIa inhibitors—including asundexian and milvexian—have emerged as potential anticoagulation therapies, based on clinical observations that patients with factor XIa deficiencies seldom present with spontaneous bleeding tendencies. The interferences associated with DOACs in routine and specialised coagulation assays are well-described; however, the interferences associated with emerging FXIa inhibitors are largely uncharacterised. Here, we briefly report the impact of asundexian and milvexian in routine coagulation assays using in vitro plasma-based systems. Asundexian and milvexian induce concentration-dependent prolongations in APTT-based assays with curvilinear regressions, which may be suitable for the measurement of pharmacodynamic effects at peak levels ex vivo. We also report differential sensitivities of APTT-based assays—particularly at higher FXIa inhibitor concentrations—highlighting the clinical need for an extensive evaluation of interferences associated with FXIa inhibitors in coagulation assays. Full article

The study investigated the biological properties of a composite material composed of poly(lactide) (PLA) and iron (Fe) produced by sputtering iron onto melt-blown poly(lactide) nonwoven fabrics. The research aimed to thoroughly understand the structure and properties of these materials and their potential applications in biomedicine. We conducted comprehensive chemical and structural analyses using techniques such as microscopic analysis, flame atomic absorption spectrometry (FAAS), and Brunauer–Emmett–Teller (BET) surface area analysis to precisely determine the properties of PLA-Fe materials. Additionally, we evaluated their impact on blood coagulation processes by measuring activated partial thromboplastin time (aPTT) and prothrombin time (PT). We also performed biological analyses on human peripheral blood mononuclear cells (PBM cells) including cell viability and DNA damage. Our results clearly demonstrate that PLA-Fe materials do not significantly influence blood coagulation mechanisms, as they only slightly prolong aPTT time and have no effect on PT. This suggests their potential in biomedical applications. Our results indicate the absence of cyto- and genotoxic properties of PLA-Fe materials against normal blood cells. In conclusion, the research findings suggest that the novel poly(lactide) and iron-sputtered nonwoven fabrics are promising tools in the field of biomedicine, offering potentially innovative therapeutic solutions for the treatment of wounds and injuries. Full article

Paeonia lactiflora Pall. (PLP) is thought to promote blood circulation and remove blood stasis. This study used blood component analysis, network pharmacology, and molecular docking to predict the mechanism of PLP in the treatment of blood stasis syndrome (BSS). PLP was processed into Paeoniae Radix Alba (PRA) and Paeoniae Radix Rubra (PRR). PRA and PRR could significantly reduce whole blood viscosity (WBV) at 1/s shear rates and could increase the erythrocyte aggregation index (EAI), plasma viscosity (PV), and erythrocyte sedimentation rate (ESR) of rats with acute blood stasis. They prolonged the prothrombin time (PT), and PRR prolonged the activated partial thromboplastin time (APTT). PRA and PRR increased the thrombin time (TT) and decreased the fibrinogen (FBG) content. All the results were significant (p < 0.05). Ten components of Paeoniflorin, Albiflorin, Paeonin C, and others were identified in the plasma of rats using ultra-high-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). A protein–protein interaction network (PPI) analysis showed that AKT1, EGFR, SRC, MAPK14, NOS3, and KDR were key targets of PLP in the treatment of BSS, and the molecular docking results further verified this. This study indicated that PLP improves BSS in multiple ways and that the potential pharmacological mechanisms may be related to angiogenesis, vasoconstriction and relaxation, coagulation, and the migration and proliferation of vascular cells. Full article

Natural and synthetic colorants present in food can modulate hemostasis, which includes the coagulation process and blood platelet activation. Some colorants have cardioprotective activity as well. However, the effect of genipin (a natural blue colorant) and synthetic blue colorants (including patent blue V and brilliant blue FCF) on hemostasis is not clear. In this study, we aimed to investigate the effects of three blue colorants—genipin, patent blue V, and brilliant blue FCF—on selected parameters of hemostasis in vitro. The anti- or pro-coagulant potential was assessed in human plasma by measuring the following coagulation times: thrombin time (TT), prothrombin time (PT), and activated partial thromboplastin time (APTT). Moreover, we used the Total Thrombus formation Analysis System (T-TAS, PL-chip) to evaluate the anti-platelet potential of the colorants in whole blood. We also measured their effect on the adhesion of washed blood platelets to fibrinogen and collagen. Lastly, the cytotoxicity of the colorants against blood platelets was assessed based on the activity of extracellular lactate dehydrogenase (LDH). We observed that genipin (at all concentrations (1–200 µM)) did not have a significant effect on the coagulation times (PT, APTT, and TT). However, genipin at the highest concentration (200 µM) and patent blue V at the concentrations of 1 and 10 µM significantly prolonged the time of occlusion measured using the T-TAS, which demonstrated their anti-platelet activity. We also observed that genipin decreased the adhesion of platelets to fibrinogen and collagen. Only patent blue V and brilliant blue FCF significantly shortened the APTT (at the concentration of 10 µM) and TT (at concentrations of 1 and 10 µM), demonstrating pro-coagulant activity. These synthetic blue colorants also modulated the process of human blood platelet adhesion, stimulating the adhesion to fibrinogen and inhibiting the adhesion to collagen. The results demonstrate that genipin is not toxic. In addition, because of its ability to reduce blood platelet activation, genipin holds promise as a novel and valuable agent that improves the health of the cardiovascular system and reduces the risk of cardiovascular diseases. However, the mechanism of its anti-platelet activity remains unclear and requires further studies. Its in vivo activity and interaction with various anti-coagulant and anti-thrombotic drugs, including aspirin and its derivatives, should be examined as well. Full article