Plasma Recalcification Time Research Articles

“Sikil pa'ak”: A proposed food from the Mayan biocultural cuisine for the prevention of metabolic syndrome

Traditional dishes play an important role in the management of diseases linked to metabolic syndrome (MS). In this context, “sikil pa'ak” (SPK), a Mayan cuisine dish made of toasted pumpkin seeds (TPS), tomato sauce with coriander (TSC), and sour orange juice (SOJ), could stand out because of the nutritional and functional properties of its ingredients. This study evaluated the sensory, nutritional, bioactive content, and functional properties of SPK and its ingredients to determine its potential in managing MS. SPK showed a water activity of 1.00 ± 0.04 and color values L* 52.74 ± 0.72, a* 12.59 ± 0.29, b* 30.16 ± 0.67. Microbiological tests confirmed the absence of contaminants. Sensorially, the flavor was the most appreciated attribute, and 29.89% rated it as “I like it very much.” Nutritionally, SPK excelled with high protein, fiber, and lipid contents with values of 24.10, 15.82, and 41.22 g/100g, respectively. The ingredients that contributed most to the content of bioactive compounds in the SPK were TPS in total phenols (TPC: 169.64 μg/mg) and total flavonoids (TFC: 234.93 μg/mg), as well as SOJ in vitamin C (65.49 μg/mg), and TSC in β-carotene (21.12 μg/mg). SPK showed high antioxidant potential (ABTS assay: 53.27%). Although SPK had a moderate effect on lipase inhibition (15.89%) and clot lysis (11.46%), it had high inhibition of α-amylase (55.21%) and plasma recalcification time (507.34 s). The SPK was sensorially appreciated and high in nutrients and bioactive compounds, showing beneficial effects against MS, especially because of TPS and SOJ. This traditional dish could be beneficial for metabolic health.

Spatiotemporally Controlled Release of Etamsylate from Bioinspired Peptide-Functionalized Nanoparticles Arrests Bleeding Rapidly and Improves Clot Stability in a Rabbit Internal Hemorrhage Model.

Achieving rapid clotting and clot stability are important unmet goals of clinical management of noncompressible hemorrhage. This study reports the development of a spatiotemporally controlled release system of an antihemorrhagic drug, etamsylate, in the management of internal hemorrhage. Gly-Arg-Gly-Asp-Ser (GRGDS) peptide-functionalized chitosan nanoparticles, with high affinity to bind with the GPIIa/IIIb receptor of activated platelets, were loaded with the drug etamsylate (etamsylate-loaded GRGDS peptide-functionalized chitosan nanoparticles; EGCSNP). Peptide conjugation was confirmed by LCMS, and the delivery system was characterized by DLS, SEM, XRD, and FTIR. In vitro study exhibited 90% drug release till 48 h fitting into the Weibull model. Plasma recalcification time and prothrombin time tests of GRGDS-functionalized nanoparticles proved that clot formation was 1.5 times faster than nonfunctionalized chitosan nanoparticles. The whole blood clotting time was increased by 2.5 times over clot formed under nonfunctionalized chitosan nanoparticles. Furthermore, the application of rheometric analysis revealed a 1.2 times stiffer clot over chitosan nanoparticles. In an in vivo liver laceration rabbit model, EGCSNP spatially localized at the internal injury site within 5 min of intravenous administration, and no rebleeding was recorded up to 3 h. The animals survived for 3 weeks after the injury, indicating the strong potential of the system for the management of noncompressible hemorrhage.

ANK-1, ANK-2, ITL-2 Polyphenols in a Dexamethasone-Induced Rat Model of Type 2 Diabetes Mellitus Treatment

Background: Diabetes mellitus (DM) is a prevalent endocrine disorder that significantly increases the risk of cardiovascular complications due to the dysfunction of the coagulation and anticoagulation systems, exacerbated by metabolic imbalances. Despite advancements in understanding DM's pathophysiology, the precise alterations in the hemostatic system remain inadequately explored, posing challenges in diagnosis and treatment. This study investigated the effects of polyphenolic compounds isolated from Hexagalloy-lglucose (ANK-1), Hepta galloyl-glucose (ANK-2), and Isatis tinctoria L. (ITL-2) on biochemical and coagulation parameters in a dexamethasone-induced type 2 diabetes mellitus (T2DM) model in rats. Methods: T2DM was induced in 25 aged white outbred rats by administering dexamethasone (0.150 mg/kg) twice daily, with a control group receiving saline. Biochemical parameters, including glucose, total protein, ALT, AST, cholesterol, and triglycerides, were measured using a semi-automatic analyzer. Coagulation parameters such as prothrombin time (PT), activated partial thromboplastin time (APTT), plasma recalcification time (RP), and fibrinogen levels were assessed using a single-channel coagulometer. Platelet aggregation was recorded using an aggregometer, and clot degradation was measured spectrophotometrically. Results: Dexamethasone administration induced significant hyperglycemia and hypercoagulation, evidenced by reduced PT, APTT, and RP, alongside increased fibrinogen levels. The administration of ITL-2 normalized all biochemical parameters, while ANK-1 and ANK-2 normalized glucose, cholesterol, and triglycerides but not ALT and AST levels. ANK-1, ANK-2, and ITL-2 demonstrated significant antithrombotic effects, with ITL-2 showing the highest efficacy, reducing clot mass and enhancing clot degradation. Conclusion: Polyphenolic compounds, particularly ITL-2, exhibit potential as therapeutic agents for managing coagulation abnormalities and hyperglycemia in T2DM. These findings highlight the importance of further research into plant-derived substances as safer and effective alternatives in diabetes management.

Procoagulant activity of red blood cell microparticles in stored packed red blood cell units and its relation to ABO blood grouping

BackgroundThroughout the storage of blood, the red cells undergo alterations known as “storage lesions,” which involve shape changes and the formation of microparticles (MPs). Studies of the formation of red cell microparticles (RMPs) emphasize the prospective application of RMPs as a quality control measure in the preparation and storage of blood components in the future. In the present study, twenty packed RBC units in citrate phosphate dextrose adenine-1 (CPDA1) were collected from volunteers and stored for 35 days. Over 35 days of storage, samples were collected at six distinct time points weekly and evaluated for the presence of RMPs. MPs were separated by the ultracentrifugation method. Electron microscopy was used to characterize the morphology and size of the isolated microparticles, and flow cytometry was performed to determine the percentage of RMPs that expressed glycophorin A (CD235a) and Annexin V antigens. RMPs' procoagulant activity (PCA) was assessed using a plasma recalcification test. RMP concentration in accordance with ABO blood grouping was assessed by using various types of donated blood groups.ResultsRMPs progressively increased over storage. The procoagulant activity (PCA) exhibited a significant increase during storage, as evidenced by a shorter plasma recalcification time (P value = 0.001). A significant negative correlation (P value = 0.001) between plasma recalcification time and Annexin V-positive microparticles, as well as a dual-positive Annexin V/CD235a population, was identified, indicating a strong correlation between the direct quantitative assay by flowcytometry and the functional assay through the PCA.ConclusionRMPs increase on storage with increased PCA. Finding ways to reduce these microparticles in packed RBC units is crucial for reducing the risk of transfusion-related coagulopathy.

Kenaf Seed Cysteine Protease (KSCP) Inhibits the Intrinsic Pathway of the Blood Coagulation Cascade and Platelet Aggregation.

Thrombosis is the key event that obstructs the flow of blood throughout the circulatory system, leading to stroke, myocardial infarction and severe cardiovascular complications. Currently, available antithrombotic drugs trigger several life-threatening side effects. Antithrombotic agents from natural sources devoid of adverse effects are grabbing high attention. In our previous study, we reported the antioxidant, anticoagulant and antiplatelet properties of kenaf seed protein extract. Therefore, in the current study, purification and characterization of cysteine protease from kenaf seed protein extract responsible for potential antithrombotic activity was undertaken. Purification of KSCP (Kenaf Seed Cysteine Protease) was carried out using gel permeation and ion exchange column chromatography. The purity of the enzyme was evaluated by SDS PAGE (Sodium Dodecyl-Sulfate Polyacrylamide Gel Electrophoresis). RP-HPLC (Reverse Phase High-Performance Liquid Chromatography), MALDI-TOF (Matrix-Assisted Laser Desorption Ionization Time-Of-Flight) and CD (Circular Dichroism techniques) were employed for its characterization. Proteolytic, fibrinolytic and kinetic study was done using spectroscopy. Plasma recalcification time, Prothrombin Time (PT), Thrombin clotting time (TCT), Activated Partial Thromboplastin Time (APTT), bleeding time and platelet aggregation studies were carried out for antithrombotic activity of KSCP. A single sharp band of KSCP was observed under both reduced and non-reduced conditions, having a molecular mass of 24.1667kDa. KSCP was found to contain 30.3% helix turns and 69.7% random coils without a beta-pleated sheet. KSCP digested casein and fibrin, and its activity was inhibited by iodoacetic acid (IAA). KSCP was optimally active at pH 6.0 at the temperature of 40°C. KSCP exhibited anticoagulant properties by interfering in the intrinsic pathway of the blood coagulation cascade. Furthermore, KSCP dissolved both whole blood and plasma clots and platelet aggregation. KSCP purified from kenaf seed extract showed antithrombotic potential. Hence, it could be a better candidate for the management of thrombotic complications.

Dynamically Cross-Linked Double-Network Hydrogels with Matched Mechanical Properties and Ideal Biocompatibility for Artificial Blood Vessels.

Vessel transplantation is currently considered the "gold standard" treatment for cardiovascular disease. However, ideal artificial vascular grafts should possess good biocompatibility and mechanical strength that match those of native autologous vascular tissue to promote in vivo tissue regeneration. In this study, a series of dynamic cross-linking double-network hydrogels and the resultant hydrogel tubes were prepared. The hydrogels (named PCO), composed of rigid poly(vinyl alcohol) (PVA), flexible carboxymethyl chitosan (CMCS), and a cross-linker of aldehyde-based β-cyclodextrin (OCD), were formed in a double-network structure with multiple dynamical cross-linking including dynamic imine bonds, hydrogen bonds, and microcrystalline regions. The PCO hydrogels exhibited superior mechanical strength, good network stability, and fatigue resistance. Additionally, it demonstrated excellent cell and blood compatibility. The results showed that the introduction of CMCS/OCD led to a significant increase in the proliferation rate of endothelial cells seeded on the surface of the hydrogel. The hemolysis rate in the test was lower than 0.3%, and both protein adsorption and platelet adhesion were reduced, indicating an excellent anticoagulant function. The plasma recalcification time test results showed that endogenous coagulation was alleviated to some extent. When formed into blood vessels and incubated with blood, no thrombus formation was observed, and there was minimal red blood cell aggregation. Therefore, this novel hydrogel tube, with excellent mechanical properties, exhibits antiadhesive characteristics toward blood cells and proteins, as well as antithrombotic properties, making it hold tremendous potential for applications in the biomedical and engineering fields.

The Prediction of Milk Whey Extract (MWE) Bioactive Compounds Based on Proximate Analysis and its Effects on Blood Coagulation: A New Approach

The present work demonstrates the milk whey extraction, proximate analysis, and biochemical characterization of bioactive compounds in MWE. Effects of MWE on plasma coagulation and platelet aggregation. Proximate analysis of MWE was done according to AOAC. Whey protein banding was confirmed in 12%, 15%, and 18% SDS-PAGEs. A quantitative analysis of bioactive compounds was done. The anticoagulant effects of MWE were tested using plasma recalcification time in both PRP and PPP, further confirmed by PT and APTT assays and in tail bleeding assays at concentrations of 0-100 µg. The non-toxic property of MWE was screened by edema, hemorrhage, and direct hemolytic activities. MWE proximate analysis showed the presence of both micro- and macronutrients. Qualitative analysis confirmed the presence of proteins and carbohydrates. A similar protein-banding pattern was observed in 12%, 15%, and 18% SDS-PAGE’s in both reduced and non-reduced conditions. MWE prolonged the clotting time of human citrated plasma, both PRP and PPP, against the control 210 sec to 770 sec, suggesting its anticoagulant property. MWE delayed the clot progress of only the APTT ratio (8.99 ± 0.09), and PT was not altered, suggesting its role in an intrinsic pathway of blood coagulation. MWE exhibited antiplatelet activity in PRP against ADP and epinephrine-induced platelet aggregation. The percentage of inhibition was 74% and 100% for ADP and epinephrine, respectively. Interestingly, MWE exhibits nontoxic properties, it does not cause hemolysis, hemorrhage, or edema. Milk whey extraction and studies confirm bioactive compounds, and these exhibit antithrombotic, antiplatelet, and non-toxic properties. Hence, MWE not only acts as a good nutritive source but also may prove tobe a therapeutic bioactive compound in the management of cardiovascular disease.

Effects of heterogeneous surface characteristics on hemocompatibility and cytocompatibility of bacterial nanocellulose

The surface properties of cardiovascular biomaterials play a critical role in their biological responses. Although bacterial nanocellulose (BNC) materials have exhibited potential applications in cardiovascular implants, the impact of their surface characteristics on biocompatibility has rarely been studied. This study investigated the mechanism for the biocompatibility induced by the physicochemical properties of both sides of BNC. With greater wettability and smoothness, the upper BNC surface reduced protein adsorption by 25 % compared with the lower surface. This prolonged the plasma re-calcification time by 14 % in venous blood. Further, compared with the lower BNC surface, the upper BNC surface prolonged the activated partial thromboplastin time by 5 % and 4 % in arterial and venous blood, respectively. Moreover, the lower BNC surface with lesser rigidity, higher roughness, and sparser fiber structure promoted cell adhesion. The lower BNC surface enhanced the proliferation rate of L929 and HUVECs cells by 15 % and 13 %, respectively, compared with the upper BNC surface. With lesser stiffness, the lower BNC surface upregulated the expressions of CD31 and eNOS while down-regulating the ICAM-1 expression - This promoted the proliferation of HUVECs. The findings of this study will provide valuable insights into the design of blood contact materials and cardiovascular implants.

New steroidal saponins from the aerial parts of Paris polyphylla var. yunnanensis and their effects on blood coagulation

Five new steroidal saponins, paripolins D-H (1–5), and 6 known compounds (6–11) were isolated from the aerial parts of Paris polyphylla var. yunnanensis. The structures of 1–5 were determined using spectroscopic analyses in conjunction with acid hydrolysis. It is for the first time to report the 12-hydroxysteroidal saponins from the genus Paris. The effect of all isolated compounds on blood coagulation was determined in vitro using the plasma recalcification time method. Compounds 1 and 2 showed potent procoagulant activity, and 5–11 exhibited significant anticoagulant activity.

Fabrication of Silk Fibroin Film with Strong Anticoagulant Property from the Synergy of Deep Eutectic Solvents and UV-Light Irradiation

As a biological macromolecule, silk fibroin (SF) has great application potential in anticoagulant. However, its complex molecular structure leads to its poor solubility, which limits the application of SF. In this study, a novel SF film with strong anticoagulant properties was fabricated through the synergistic effect of LiBr deep eutectic solvents (DESs) and UV-light irradiation. Under the optimum conditions, the plasma recalcification time of SF film was 182 s, the hemolysis rate was 3.93%, and the dynamic coagulation time remained at a high level up to 60 min. The synergistic effect of UV-light and DES can promote the enhancement of hydrophilicity and the decrease of surface roughness of SF films, and thus the anticoagulant properties of the films were enhanced. The structure change, wettability, and roughness of SF films were characterized by XRD, FTIR, contact angle, and AFM. In the meantime, the wavelength of the UV-light and the irradiation time have an effect on the anticoagulant properties of the SF films. Furthermore, the improvement of the anticoagulant properties of SF films exhibits good universality for different LiBr DESs. The study provides a new direction for the preparation of strong anticoagulation SF films, as well as its application in life sciences.

Water-Induced Expanded Bilayer Vascular Graft with Good Hemocompatibility and Biocompatibility.

Shape memory polymer (SMP) vascular grafts are promising interventional vascular grafts for cardiovascular disease (CAD) treatment; However, hemocompatibility and biocompatibility, which are the critical issues for the SMP vascular grafts, are not systematically concerned. Furthermore, the water-induced SMP grafts are more convenient and safer than the thermally induced ones in case of the bioapplication. Herein, in this work, the new water-induced expanded bilayer vascular graft with the inner layer of crosslinked poly(ε-caprolactone) (cPCL) and the outer layer of water-induced SMP of regenerated chitosan/polyvinyl alcohol (RCS/PVA) are prepared by hot pressing and programming approaches. The results show that the inner and outer layer surfaces of the prepared grafts are smooth, and they exhibit good interfacial interaction properties. The bilayer grafts show good mechanical properties and can be expanded in water with a diameter expansion of ≈30%. When compared with commercial expanded polytetrafluoroethylene (ePTFE), the bilayer graft shows better hemocompatibility (platelet adhesion, hemolysis rate, various clotting times, and plasma recalcification time (PRT)) and in vitro and in vivo biocompatibility, which thus is a promising material for the vascular graft.

Development of substrate-independent heparin coating to mitigate surface-induced thrombogenesis: efficacy and mechanism.

Heparin coatings are widely applied on blood-contact materials to reduce the use of anticoagulants during blood treatment. However, the previous heparin coatings formed via covalent binding or electrostatic bonding commonly require complex surface premodification, and the blood coagulation pathway was significantly inhibited to potentially increase the bleeding risk. This contradicts the intended purpose and deviates from the anticoagulation mechanism of the heparin coatings. Herein, we present a facile and substrate-independent coating, achieved through the co-deposition of dopamine/chitosan followed by electrostatic interaction between heparin and the immobilized chitosan, which could be prepared within 1 hour. This coating prolonged the plasma re-calcification time (PRT) to over 60 minutes, effectively preventing surface-induced thrombosis. Favorable hemocompatibility was reflected in a hemolysis ratio of less than 2%, low levels of platelet adhesion and activation, and low levels of fibrinogen adhesion. We also systematically elucidate the anticoagulant mechanism of the coating, demonstrating why the coating can prevent thrombogenesis without the bleeding risk. Our work not only offers a promising and readily available heparin coating for blood-contact materials, but more importantly, the mechanism exploration supports the practical feasibility of heparin coating in various applications.

Bat-derived oligopeptide LE6 inhibits the contact-kinin pathway and harbors anti-thromboinflammation and stroke potential.

Thrombosis and inflammation are primary contributors to the onset and progression of ischemic stroke. The contact-kinin pathway, initiated by plasma kallikrein (PK) and activated factor XII (FXIIa), functions bidirectionally with the coagulation and inflammation cascades, providing a novel target for therapeutic drug development in ischemic stroke. In this study, we identified a bat-derived oligopeptide from Myotis myotis (Borkhausen, 1797), designated LE6 (Leu-Ser-Glu-Glu-Pro-Glu, 702 Da), with considerable potential in stroke therapy due to its effects on the contact kinin pathway. Notably, LE6 demonstrated significant inhibitory effects on PK and FXIIa, with inhibition constants of 43.97 μmol/L and 6.37 μmol/L, respectively. In vitro analyses revealed that LE6 prolonged plasma recalcification time and activated partial thromboplastin time. In murine models, LE6 effectively inhibited carrageenan-induced mouse tail thrombosis, FeCl 3-induced carotid artery thrombosis, and photochemically induced intracerebral thrombosis. Furthermore, LE6 significantly decreased inflammation and stroke injury in transient middle cerebral artery occlusion models. Notably, the low toxicity, hemolytic activity, and bleeding risk of LE6, along with its synthetic simplicity, underscore its clinical applicability. In conclusion, as an inhibitor of FXIIa and PK, LE6 offers potential therapeutic benefits in stroke treatment by mitigating inflammation and preventing thrombus formation.

Screening and verification of hemostatic effective components group of Panax Notoginseng based on spectrum-effect relationships

Ethnopharmacological relevancePanax Notoginseng (PN) can disperse blood stasis, hemostasis, and detumescence analgesic, which can be used for hemoptysis, hematemesis and another traumatic bleeding, and it is known as “A miracle hemostatic medicine”. Studies show that the chemical composition of PN is relatively comprehensive, however, its hemostatic active ingredients have not been fully clarified. Aim of studyThis study aimed to clarify the hemostatic effective components group (HECG) of PN, provide a foundation for the assessment of PN's quality and its comprehensive development, and for further studies on the pharmacodynamic material basis of other Traditional Chinese Medicines (TCMs). Materials and methodsUPLC-MS was used to establish the fingerprint and identify the common peaks in 44 batches of PN extracts (PNE). In addition, the plasma recalcification time and in vitro coagulation time were measured. For spectrum-effect analysis, bivariate correlation analysis (BCA) and partial least squares regression analysis (PLSR) were used to screen the hemostasis candidate active monomers of PN. The monomers were prepared by combining several preparative chromatography techniques. The efficacy was verified by plasma recalcification time, in vitro coagulation time, and a rat model of gastric hemorrhage. ResultsA total of 30 common peaks and hemostatic efficacy indexes of 44 batches of PNE were obtained. A total of 18 components were positively correlated with the comprehensive coagulation index by two statistical methods. Six and eleven monomers were obtained respectively by chromatographic preparation and procurement, and one monomer was eliminated due to preparation difficulty and other reasons. Seven active monomers with direct hemostatic effect and one active monomer with synergistic hemostatic effect were screened through plasma recalcification time, and their combinations were used as candidate HECG for hemostatic effect verification. The results of in vitro experiments showed that plasma recalcification time and in vitro coagulation time were significantly reduced (P < 0.05) in the HECG group, compared to the PNE group. The results of in vivo experiment also indicated that the hemostatic effect of HECG was comparable to that of PNE and PN powder. ConclusionThe composition and efficacy of the HECG of PN were screened and verified using the spectral correlation method and in vivo and in vitro efficacy verification; the HECG included Dencichine, Ginsenoside Rg1, Ginsenoside Rd, Ginsenoside Rh1, Ginsenoside F1, Notoginsenoside R1, Notoginsenoside Ft1 and Notoginsenoside Fe. These results laid a foundation for the quality evaluation of PN and provided a reference for the basic research of pharmacodynamic material basis of other TCMs.

TiO2-doped bilirubin-imprinted polydopamine cored with Fe3O4 nanoparticle for bilirubin selective adsorptivity enhancement

As one of the essential treatments for bilirubin removal in clinical, hemoperfusion of bio-sorbents required efficacy, efficiency and biosafety. Dopamine of biocompatibility and spontaneous polymerization was herein proposed for the bilirubin-imprinted adsorptive polymers of selectivity, doped with TiO2 of adsorptivity and Fe3O4 as fixed magnets. Via electrostatic reaction and hydrogen bonding, the magnetic sorbents composed of polydopamine doped with TiO2 removed bilirubin of 263.5 mg·g−1 non-homogeneously during 240 min. With a competitive adsorption coefficient (tN) of 10.4, the magnetic sorbents also exhibited the decent blood compatibility of acceptable hemolysis ratio and plasma recalcification time. Prospectively, the TiO2-doped bilirubin-imprinted PDA cored with Fe3O4 was potentially for bilirubin selective removal via hemoperfusion in clinic.

Enantioselective Synthesis, Computational Molecular Docking and In Vitro Anticoagulant Activity of Warfarin-based Derivatives

Abstract: Warfarin containing a 4-hydroxycoumarin moiety possesses excellent anticoagulant activity, with the (S) enantiomer being the eutomer. The present work is designed to synthesize warfarin based derivatives enantioselectivity to explore their anticoagulant potential. The substituted chalcones were reacted with 4-hydroxycoumarin in the presence of the chiral organocatalyst 9-amino-9-deoxyepiquinine to afford warfarin-based analogues 5a- 5k. The structures of synthesized compounds 5a-5k were confirmed by Fourier transform infrared spectroscopy (FTIR), proton nuclear magnetic resonance spectroscopy (1H NMR), carbon-13 nuclear magnetic resonance spectroscopy (13C NMR) and electron ionization mass spectroscopy (EIMS) data. The enantiomeric excess (ee) has been found in the range of 16-99% as determined by chiral high-performance liquid chromatography (HPLC) analysis. The in vitro anticoagulant activity of the products 5a-5k was evaluated by plasma recalcification time (PRT) method, and it was found that most of the derivatives showed good anticoagulant activity, specifically compound 5b exhibited excellent results compared to that of warfarin. Compound 5b displayed an IC50 value of 249.88 μM, which is better than that of warfarin (IC50 408.70 μM). The molecular docking studies have been performed against vitamin K epoxide reductase with PDBID 3kp9. The synthesized compounds bind well in the active binding site of the target enzyme. The derivative 5b showed π-π stacking interactions with the amino acid phenylalanine (Phe 114). The antimicrobial activity of synthesized compounds has also been evaluated, and results showed moderate antimicrobial activity. Based on our results, it is proposed that derivative 5b may act as a lead compound to design more potent anticoagulant derivatives.

RATIONALE FOR PRESCRIBING ANTICOAGULANT THERAPY IN THE COMPLEX TREATMENT OF PURULENT-NECROTIC COMPLICATIONS OF DIABETIC FOOT SYNDROME

Abstract. This article is based on the study of hemostasis indicators in 25 patients with diabetic foot syndrome. Diabetic foot syndrome (DFS) develops against the background of diabetic neuropathy, micro- and macroaniopathy, osteoarthropathy, which lead to frequent traumatization of the soft tissues of the foot, infection with the subsequent development of purulent-necrotic processes on the foot, and in the course of progression leads to high limb amputations. According to the international federation, which deals with the problem of diabetes mellitus (DM), the 21st century is considered a pandemic of DM. It should be noted that the number of patients with diabetes is increasing every year. Patients with diabetes are characterized by an atherothrombotic state caused by multiple disorders in the hemostasis system. Hemostatic disorders in patients with diabetes are complex and include activation of natural mechanisms of the coagulation system, inhibition of fibrinolytic activity, and various platelet dysfunctions.&#x0D; The purpose of the study is to substantiate the appointment of anticoagulation therapy in patients with SDS.&#x0D; Materials and methods. On the basis of KNP KMKL No. 6 in Kyiv in 2023, for a period of 6 months, 69 patients with SDS were treated. The state of indicators of the coagulation link of the hemostasis system in 49 people was studied. At the same time, 26 patients had purulent-necrotic complications of the neuroischemic form of SDS, 23 people were healthy (comparison group). The patients were divided according to the article and age, according to the duration of diabetes mellitus, and according to the stages according to Wagner. The following indicators were determined for patients: prothrombin index (PTI), plasma recalcification time, fibrinogen, B-naphthol test, ethanol test, Willebrand factor, endothelin-1, coagulation factor (VIII), fibronectin.&#x0D; The results of the study and their discussion: Based on the study, 61.5 % of patients with SDS are of working age. Of them, 57.7 % are men, 42.3 % are women. The largest number of patients with SDS is observed in the group of patients who have had diabetes for more than 10 years - 57.7 %, and the largest number of patients with stage III-IV according to Wagner - 88.5 %. In patients with diabetes with neuroischemic form of SDS, the PTI level is increased by 28.8 % and the prothrombin time is shortened by 21 %, which indicates an increase in the activity of external blood clotting mechanisms. An increase in fibrinogen by 29.7 % indicates an increase in the activity of coagulation hemostasis. Willebrand factor increased by 34.5 %, coagulation factor (VIII) increased by 20.3 %, fibronectin increased by 83.01 %, which indicates an increase in aggregation and procoagulant processes, which in turn increases thrombus formation.&#x0D; Conclusion. Thus, with diabetic foot syndrome, hypercoagulation is observed, which leads to an increase in the level of thrombus formation, which justifies the appointment of anticoagulant therapy in the complex treatment of diabetic foot syndrome.

Evaluation of PTFPMS membrane towards application in artificial oxygenation

The tradeoff between efficient gas exchange and blood leakage as well as thrombosis caused by contact between blood and membrane are existing challenges in extracorporeal membrane oxygenation (ECMO). Herein, poly (3,3,3-trifluoropropyl) methylsiloxane/polyetherimide (PTFPMS/PEI) composite membranes, with a thin PTFPMS dense layer for functional CO2 and O2 exchange coated on microporous PEI substrate membrane for mechanical support, were synthesized, characterized and evaluated for potential application as oxygenation membrane. Characterization data from scanning electron microscopy with energy dispersive spectroscopy, thermogravimetric analysis, contact angle and zeta potential measurements confirmed that PTFPMS/PEI composite membrane with different coating concentrations were successfully prepared, and the composite membranes obtained exhibited thermal stability, surface hydrophobicity as well as surface electronegativity. Analysis on gas permeance and hemocompatibility performance demonstrated that the composite membranes displayed effective gas transfer and good blood compatibility in terms of hemolysis rate, protein adsorption, platelet adhesion as well as plasma recalcification time. More notably, the membrane also displayed excellent long term anti-fouling performance. In summary, the PTFPMS/PEI composite membrane showed impressive application potential for use as oxygenation membrane in ECMO.

Biocompatibility assessment of Rutin and PEG loaded novel nanoceria on human erythrocytes and human myeloid leukemia (U937) cells

Nanomaterials are increasingly being used, prompting extensive research into nanomedicine. The use of nanomaterials is constrained by the absence of comprehensive toxicity profiles, despite their impressive diagnostic and therapeutic outcomes in in vitro and in vivo models. The compatibility of the bioconjugate nanoparticles, we fabricated a novel Rutin (Rut) and polyethylene glycol (PEG) loaded cerium oxide nanoparticles (Rut-PEG-CeO2NPs). The investigation of antibacterial activity of cerium oxide nanoparticles (CeO2NPs), Rut, Rut-PEG-CeO2 NPs and antibiotic was examined against Gram-positive and Gram-negative pathogens using well-diffusion and time-kill assays. The fabricated Rut-PEG-CeO2NPs has demonstrated broad spectrum nature resulting in significant cell damage at the target site when compared to CeO2NPs and Rut. The haemolytic effect of Rut-PEG-CeO2NPs on erythrocytes from healthy donor (HDE) and diabetic donor (DDE) was evaluated using the haemolysis assay against CeO2NPs. The results exhibit that the percentage of haemolysis was more in DDE compared to that of HDE for CeO2NPs as well as Rut-PEG-CeO2NPs. The study of anticoagulation activity using plasma recalcification time method which shows the clotting time of platelet rich plasma (PRP) 280–290 s and platelet poor plasma (PPP) 245–318 s. Besides that, the anti-angiogenetic property was evaluated using chick embryo chorioallantoic membrane (CAM) assay which showed the remarkable anti-angiogenic effect of Rut-PEG-CeO2NPs on the chick embryo. Additionally, the in vitro cytotoxicity of Rut-PEG-CeO2NPs was tested on human myeloid leukemia cells (U937) against Rut and CeO2NPs. The cytotoxicity results depicted that Rut-PEG-CeO2NPs shows high percent of cell viability compared to that of CeO2NPs and Rutin. These results suggests that the fabricated Rut-PEG-CeO2NPs has novel approach towards the therapeutic applications which is potentially due to the loading of Rut on the CeO2NPs on the surface to form Rut-PEG-CeO2NPs.

In Vitro Anticoagulant Activity of Crude Protease of Bacillus tequilensis HSFI-5

obtained from the fermented intestine of Holothuria scabra (sand sea cucumber). Strain HSFI-5 had been reported to be able to produce proteases, which had shown several characteristics of an antithrombotic agent, i.e., fibrinolytic and clot-lysis activities. However, its anticoagulation activity test is yest to be done. This study aimed to determine the anticoagulant activity of the crude protease HSFI-5 in vitro. The study design was a completely randomized design with a sample size of 90 calculated using the Federer formula. The material used was crude protease from B. tequilensis in skim milk broth. Prothrombin time (PT), activated partial thromboplastin time (aPTT), and plasma recalcification time (PRT) were carried out to test the anticoagulant activity. Citrated platelet poor plasma samples were divided into positive control, normal control, direct examination with crude enzyme in volumes of 50 and 100 µL and pre-incubation at 37ºC for 5, 10, and 15 min with crude enzyme volumes of 50 and 100 µL. The data normality was tested with the Kolmogorov-Smirnov test and the different tests were analyzed by one-way ANOVA with the Post hoc LSD test. The results of one-way ANOVA both on PT, aPTT, and PRT examinations showed that there was a significant difference between the treatment groups (p&lt;0.05). The longest results of PT, aPTT, and PRT are positive controls, and the shortest results are normal controls for PT, and 15’ 50 group for aPTT and PRT. It is clear that crude protease B. tequilensis HSFI-5 exhibits anticoagulant as well as thrombolytic action, raising the possibility that it could function as an antithrombotic drug.