Fibrinolytic Research Articles

Plasmin generation analysis in patients with bleeding disorder of unknown cause

AbstractBleeding disorder of unknown cause (BDUC) is a diagnosis of exclusion after evaluation of plasma coagulation and platelet function. Patients with BDUC (n = 375) recorded in the Vienna Bleeding Biobank were analyzed in comparison with healthy controls (HCs; n = 100) in this case-control study. Plasmin generation (PG) parameters were analyzed using calibrated fluorescence detection in citrated plasma. Turbidimetric plasma clot formation/lysis of 293 (78%) patients with BDUC and confocal microscopy of clots from representative patients with BDUC (n = 6) and HCs (n = 9) were assessed. In the PG analysis, patients with BDUC exhibited lower velocity and peak plasmin levels but a higher endogenous plasmin potential than HCs. Peak plasmin levels correlated with maximum clot absorbance but not with clot lysis time. Clot absorbance is an indicator of clot fiber density. Confocal microscopy analysis revealed a tendency towards thicker fibers in clots of patients with BDUC, which negatively correlated with peak plasmin (r = −0.561; P = .030). Peak plasmin correlated weakly with factor XIII, but not with other fibrinolytic factors (alpha2-antiplasmin, thrombin activatable fibrinolysis inhibitor, or plasminogen activator inhibitor 1) or bleeding severity. A model comprising fibrinogen and parameters of PG yielded high predictive power in discriminating between patients with BDUC and HCs across a fivefold stratified cross validation (80% of data; mean area under the curve [AUC], 0.847). The model generalized well to unseen data (20% of data; AUC, 0.856). Overall, patients with BDUC counterintuitively exhibited reduced peak plasmin levels, potentially related to altered clot structure.

Ocimum basilicum L. leaves extract-mediated green synthesis of MnO NPs: Phytochemical profile, characterization, catalytic and thrombolytic activities

ObjectiveThe goal of the present work was the identification and quantification of the phenolic profile of sweet basil using LC-ESI-MS and to evaluation the ability of this plant to synthesise manganese nanoparticles MnONPs and evaluation of their catalytic and thrombolytic activities. Material and methodsStandard procedures were used to extract bioactive molecules and test qualitative phytochemical substances. Phenolic compounds were identified and quantified using LC-ESI-MS. Furthermore, visual observation, UV–Vis and FT-IR spectroscopy, scanning and transmission electron microscopes (SEM and TEM), and XRD technique were used to verify the green synthesis of ObE-MnO NPs. Dye degradation of ObE-MnO NPs was studied using photocatalytic and sonocatalytic activities. Moreover, in vitro thrombolytic activity was evaluated by clot lysis. ResultsResults of phytochemical essays showed that the aqueous extract of Ocimum basilicum L. is very rich in different chemical compounds that reduce and stabilize NPs. The results of the characterization of MnONPs show that the formation of these particles was proven by UV–Vis spectrum with the absorption peak at 405 nm, and also by FTIR spectrum at 470-522 cm−1. The study also confirmed that the MnONPs were spherical in shape with an average size of 6.52 ± 0.88 nm using TEM and SEM analysis, and that their size was 13.4 nm, tetragonal, and crystalline by XRD data. Results of catalytique activity show at a time of 240 min, an efficacy level of 70.18% and 58.90% for the photocatalytic and sonocatalytic activities. Ocimum basilicum L. extract and ObE-MnO NPs demonstrated a significant higher percentage of clot lysis by 93.33% and 94.73% respectively. ConclusionIn conclusion, the synthesized NPs made with basil extract were satisfactorily characterized utilizing a range of analytical techniques. ObE-MnO NPs also exhibit efficient catalytic and thrombolytic activities, qualifying their high degree of safety for usage in the environmental, pharmacological and medicinal domains.

Safety and Efficacy Comparison of Tenecteplase and Alteplase for Clinically Suspected Large Vessel Occlusion Strokes without Thrombectomy

Introduction: Tenecteplase is a thrombolytic with higher fibrin affinity and is potentially better in clot lysis. A higher spontaneous recanalisation rate for large vessel occlusion (LVO) strokes had been shown in comparison studies with alteplase. Results of the LVO studies reflect the composite effect of the thrombolytic and thrombectomy, as patients would be treated by thrombectomy had they not been recanalised by intravenous thrombolysis alone. Thrombectomy is not readily available in many parts of the world. Our study aimed to compare the outcomes of suspected LVO patients treated with tenecteplase versus alteplase only, without the confounding effect of thrombectomy. Methods: This is a retrospective review. Data of patients given tenecteplase from May 2020 to August 2023 and those given alteplase 0.9 mg/kg from January 2019 to August 2023 were retrieved. Due to fluctuation in supply of tenecteplase during the COVID pandemic, some LVO patients were given alteplase. Patients with anterior circulation, clinically suspected LVO strokes (defined as National Institutes of Health Stroke Scale (NIHSS) score ≥6, plus cortical signs or hyperdense vessel sign), with thrombolysis given within 4.5 h of stroke onset were analysed. Patients with thrombectomy done were excluded. Safety and efficacy outcomes were compared. Results: There were 245 tenecteplase-treated patients treated between May 1, 2020, and August 31, 2023, and 732 patients were treated with alteplase between January 1, 2019, to August 31, 2023. Out of these, 148 tenecteplase patients and 138 alteplase 0.9 mg/kg patients fulfilled the study criteria. The symptomatic intracerebral haemorrhage rate was non-significantly lower in the tenecteplase group (2.1% vs. 5.8%, p = 0.13). There were no significant differences in the rate of ≥8-point NIHSS improvement (23.6% vs. 23.7%, p = 1) or the ≥4-point improvement (40.5% vs. 40.7%, p = 1) at 24 h. At 3 months, 21.6% of tenecteplase patients had good functional outcome (modified Rankin scale [mRS] 0–2), compared to 26.3% in the alteplase group (p = 0.40). Conclusion: In this pragmatic study of clinically suspected anterior circulation LVO patients without thrombectomy, outcome solely reflects the effects of tenecteplase. Tenecteplase showed comparable safety and efficacy to alteplase, but the result should be interpreted with caution in view of its small sample size and non-randomised study design.

The islet tissue plasminogen activator/plasmin system is upregulated with human islet amyloid polypeptide aggregation and protects beta cells from aggregation-induced toxicity

Aims/hypothesisApart from its fibrinolytic activity, the tissue plasminogen activator (tPA)/plasmin system has been reported to cleave the peptide amyloid beta, attenuating brain amyloid deposition in Alzheimer’s disease. As aggregation of human islet amyloid polypeptide (hIAPP) is toxic to beta cells, we sought to determine whether activation of the fibrinolytic system can also reduce islet amyloid deposition and its cytotoxic effects, which are both observed in type 2 diabetes.MethodsThe expression of Plat (encoding tPA) and plasmin activity were measured in isolated islets from amyloid-prone hIAPP transgenic mice or non-transgenic control islets expressing non-amyloidogenic mouse islet amyloid polypeptide cultured in the absence or presence of the amyloid inhibitor Congo Red. Plat expression was also determined in hIAPP-treated primary islet endothelial cells, bone marrow-derived macrophages (BMDM) and INS-1 cells, in order to determine the islet cell type(s) producing tPA in response to hIAPP aggregation. Cell-free thioflavin-T assays and MS were used to respectively monitor hIAPP aggregation kinetics and investigate plasmin cleavage of hIAPP. Cell viability was assessed in INS-1 beta cells treated with hIAPP with or without plasmin. Finally, to confirm the findings in human samples, PLAT expression was measured in freshly isolated islets from donors with and without type 2 diabetes.ResultsIn isolated islets from transgenic mice, islet Plat expression and plasmin activity increased significantly with the process of amyloid deposition (p≤0.01, n=5); these effects were not observed in islets from non-transgenic mice and were blocked by Congo Red (p≤0.01, n=4). In response to hIAPP exposure, Plat expression increased in BMDM and INS-1 cells vs vehicle-treated cells (p≤0.05, n=4), but not in islet endothelial cells. Plasmin reduced hIAPP fibril formation in a dose-dependent manner in a cell-free system, and restored hIAPP-induced loss of cell viability in INS-1 beta cells (p≤0.01, n=5). Plasmin cleaved monomeric hIAPP, inducing a rapid decrease in the abundance of full-length hIAPP and the appearance of hIAPP 1–11 and 12–37 fragments. hIAPP 12–37, which contains the critical amyloidogenic region, was not toxic to INS-1 cells. Finally, PLAT expression was significantly increased by 2.4-fold in islets from donors with type 2 diabetes (n=4) vs islets from donors without type 2 diabetes (n=7) (p≤0.05).Conclusions/interpretationThe fibrinolytic system is upregulated in islets with hIAPP aggregation. Plasmin rapidly degrades hIAPP, limiting its aggregation into amyloid and thus protecting beta cells from hIAPP-induced toxicity. Thus, increasing islet plasmin activity might be a strategy to limit beta cell loss in type 2 diabetes.Graphical

The hemostatic activity and Mechanistic roles of glucosyloxybenzyl 2-isobutylmalate extract (BSCE) from Bletilla striata (Thunb.) Rchb.f. in Inhibiting pulmonary hemorrhage

BackgroundHemorrhagic events cause numerous deaths annually worldwide, highlighting the urgent need for effective hemostatic drugs. The glucosyloxybenzyl 2-isobutylmalates Control Extract (BSCE) from the orchid plant Bletilla striata (Thunb.) Rchb.f. has demonstrated significant hemostatic activity in both in vitro and in vivo studies. However, the effect and mechanism of BSCE on non-traumatic bleeding remain unclear. MethodsPulmonary hemorrhage was induced in 40 Sprague-Dawley rats by administering Zingiber officinale Roscoe. for 14 days. These rats were then randomly divided into five groups: model (Mod), positive control (YNBY), and BSCE low, medium, and high-dose groups. An additional 8 rats served as the control group (Con). The BSCE groups received different doses of BSCE for 10 days, while the YNBY group received Yunnan Baiyao suspension. The effects on body weight, food and water intake, red blood cell count (RBC), hemoglobin concentration (HGB), lung tissue pathology, platelet count, coagulation parameters, and fibrinolytic system markers were evaluated. Network pharmacology and molecular docking analyses were also conducted to identify potential targets and pathways involved in BSCE's effects. ResultsBSCE treatment significantly improved body weight, food intake, and water consumption in rats with pulmonary hemorrhage. RBC and HGB levels increased significantly in the BSCE medium and high-dose groups compared to the Mod group (P < 0.05). Pathological examination revealed that BSCE reduced lung tissue hemorrhage and inflammation, with improvements in alveolar structure. BSCE also positively affected platelet count, thrombin time (TT), activated partial thromboplastin time (APTT), fibrinogen (FIB) levels, and fibrinolytic markers (D-dimer, PAI-1, and t-PA). Network pharmacology and molecular docking identified key targets such as MMPs, CASPs, and pathways including IL-17 and TNF signaling, suggesting BSCE's involvement in hemostasis and anti-inflammatory processes. ConclusionsBSCE exhibits significant hemostatic and protective effects on Z.officinale-induced pulmonary hemorrhage in rats by improving hematological parameters, reducing lung tissue damage, and modulating the coagulation and fibrinolytic systems. The study provides evidence supporting the potential of BSCE as a therapeutic agent for hemorrhagic diseases, with its efficacy linked to multi-target and multi-pathway interactions.

Age-related characteristics of the content and activity of certain components of the blood fibrinolytic system in cases of uterine cancer

Uterine cancer (UC) is the third most common cancer in women in Russia. Knowledge of age-specific features of the fibrinolytic system in patients with UC can be useful in terms of improving its pathogenetic therapy. Aim of the study was to investigate the peculiarities of the content and activity of some components of the fibrinolytic system in the blood of UC patients of different age groups. Material and methods. 30 healthy women (donors) and 56 patients with uterine cancer T1a-2N0M0, adenocarcinoma G1–G3 were divided into subgroups according to age: reproductive, perimenopause and menopause. ELISA of urokinase (u-PA), its receptor (u-PAR) and its inhibitor (PAI-1), tissue-type plasminogen activator (t-PA) content were performed in blood collected before treatment. Results. In reproductive UC patients, inhibitor u-PA (PAI-1) activity increased 14-fold (hereinafter the difference is statistically significant, p &lt; 0.05) and content increased 2.9-fold, while receptor u-PA (u-PAR) level decreased 1.7-fold compared to reproductive donors. Perimenopausal UC patients showed a 3.3-fold increase in PAI-1 content and a 6.3-fold increase in PAI-1 activity compared to perimenopausal donors, t-PA concentration decreased 1.3-fold relative to donors and was 1.9-fold lower than in reproductive UC patients. In menopausal patients with UC, the activity and content of PAI-1 enhanced 5.5-fold and 4.5-fold, respectively, compared to donors. Additionally, they were 2.1-fold and 1.2-fold lower than in reproductive UC patients. The activity of u-PA increased 2.6-fold, reaching the values of reproductive UC patients. The activity of t-PA was 1.3-fold higher than in donors, but did not differ from the activity in other RTM patients, while the level of t-PA occupied an intermediate position between the corresponding indices in young UC patients (it was 1.4-fold lower) and perimenopausal UC patients (it was 1.4-fold higher). Conclusions. The development of UC is accompanied by an imbalance of components of the fibrinolytic system in the blood, depending on the age of women, with a minimum spectrum of changes in the indicators in reproductive patients and maximum - in menopause patients, which indicates the pathognomonicity of these factors in PTM and requires an individual approach to the management of such patients.

Association of periodontitis with cardiometabolic and haemostatic parameters

ObjectiveTo investigate the association between periodontitis and cardiometabolic and haemostatic parameters.Materials and methodsBetween 2014 and 2019, 54 individuals needing full mouth extraction, and 50 control individuals, were recruited for a combined cross-sectional (individuals versus controls) and longitudinal (individuals before and after extraction) study. Periodontitis severity was measured using the periodontal inflamed surface area (PISA). Blood was drawn to measure the haemostatic (Factor VIII, von Willebrand factor [VWF], endogenous thrombin potential, d-dimer, clot lysis time) and cardiovascular risk (C-reactive protein [CRP], lipid profile) parameters, prior to and 12 weeks post-extraction. The results were analysed group-wise.ResultsThe mean VWF and CRP levels were higher and the high-density lipoprotein levels were lower in the individuals prior to extraction compared to the controls. The VWF was significantly correlated with the PISA (a 21% unit increase in VWF per 1000 mm2 increase in PISA, 95%CI: 6–36%, p = 0.01). The other analyses were comparable between the individuals and controls, and did not change in the individuals after the extraction.ConclusionVWF levels are associated with periodontitis severity; they do not improve after full-mouth extraction. Severe periodontitis in control individuals does not induce substantial changes in their haemostatic or inflammatory systems.Clinical relevanceTreatment of periodontitis has been shown to improve the cardiometabolic blood profile of patients with established cardiometabolic disease. However, whether periodontitis treatment improves cardiometabolic and haemostatic profiles in people without cardiometabolic disease is uncertain.

Fibrinolysis is impaired in patients with primary immune thrombocytopenia

BackgroundPatients with primary immune thrombocytopenia (ITP) have an increased risk of thrombosis, which may be due to altered fibrinolysis. ObjectivesTo elucidate the clinical impact of delayed fibrinolysis in ITP patients. MethodsA turbidimetric clot formation and lysis assay and a fluorometric plasmin generation (PG) assay were performed, and levels of plasminogen activator inhibitor-1 (PAI-1), tissue plasminogen activator (tPA), tPA-PAI-1 complexes, α2-antiplasmin, thrombin activatable fibrinolysis inhibitor, and D-dimer were assessed in 86 adult primary ITP patients and 78 healthy controls (HCs). ResultsITP patients showed significantly delayed clot formation, increased clot density, and prolonged clot lysis time (CLT) compared with HCs, with a median (IQR) CLT of 28.0 (13.7-34.7) minutes in patients and 17.3 (12.0-28.0) minutes in HCs, while in the PG assay, only the lag time was prolonged. In ITP patients compared with controls, PAI-1 was higher (1.2 [0.8-2.6] vs 1.1 [0.6-2.1] U/mL) and tPA antigen and activity were lower (tPA antigen: 2.6 [1.1-4.4] vs 3.7 [3.2-4.7] ng/mL; tPA activity ≤ 0 U/mL: 26% vs 7%). TPA-PAI-1 complex levels were positively associated with CLT in multiple linear regression analysis (β = 0.241; P = .019), whereas PG parameters were not associated with CLT. Six patients who developed thrombosis during follow-up had higher levels of tPA-PAI-1 complexes. ConclusionProlonged CLT and delayed onset of PG may indicate a hypofibrinolytic tendency in ITP patients, as also indicated by high PAI-1 and low tPA levels. No association was found between fibrinolytic potential and the bleeding phenotype, whereas higher tPA-PAI-1 complex levels were associated with prolonged CLT and increased in patients with future thrombosis.

SERPINE1 and MTHFR genetic variants in patients with embolic stroke of undetermined source: links with fibrin clot properties.

The SERPINE1 c.-820G (4_5), MTHFR gene variants, and unfavourably altered fibrin clot features, have been suspected to be associated with embolic stroke of undetermined source (ESUS). We investigated the SERPINE1 c.-820G (4_5) gene variants alone and coexisting with MTHFR c.665C > T and c.1286A > C gene variants in relation to thrombophilic factors and plasma fibrin clot properties in Polish patients with ESUS. Unrelated consecutive patients with ESUS (n = 206) were genotyped by TaqMan assay. Thrombophilia screening was performed four weeks or more after a thrombotic event while off oral anticoagulation. Factor VIII (FVIII) activity was determined by a coagulometric assay, while lipoprotein(a) was determined using immunoturbidimetry. We determined fibrin clot permeability (Ks) and clot lysis time (CLT). Apparently healthy individuals without a family history of stroke or venous thromboembolism (n = 30), and patients with a history of atrial fibrillation (n = 25) or carotid artery disease-related stroke (n = 21), served as controls. Among ESUS patients, the SERPINE1 c.-820G (4_5) minor allele frequency was 0.57. There were no differences in common factors associated with thrombophilia among ESUS patients regarding SERPINE1 variants. The overall prevalence of FVIII > 150IU/dL was 26% (n = 53) and elevated FVIII predominated in SERPINE1 variants carriers (n = 45; 84.9%), including 36 (68%) carriers of MTHFR variant. Moreover, 4.3-fold higher Lp(a) levels along with 50% reduced Ks and 46% prolonged CLT were found in patients with mutant SERPINE1 combined with mutant homozygotes in the MTHFR c.665C > T variant compared to the wild type SERPINE1 combined with mutant homozygotes in the MTHFR c.665C >T (P < 0.001). The SERPINE1 c.-820G (4_5) variants carriers have increased FVIII levels, while the SERPINE1 c.-820G (4_5) mutant homozygotes coexisting with MTHFR c.665C > T have more prothrombotic fibrin clot features and elevated Lp(a). Our study underlines the cumulative effect of genetic risk factors in patients with ESUS that might require specific antithrombotic therapy.

Real-Time Monitoring of Lower Limb Oxygen Saturation in Children with Indwelling Catheters Using Near-Infrared Spectroscopy to Assess the Risk of Deep Vein Thrombosis and Nursing Strategies

Objectives: To analyze the risk of deep vein thrombosis (DVT) in children with indwelling catheters through near-infrared spectroscopy (NIRS) detection of lower limb crSaO2, and to implement corresponding nursing countermeasures against the risk. Methods: Using a convenience sampling method, 80 children with indwelling catheters were selected from the first pediatric intensive care unit of our hospital between January 2022 and December 2022. They were divided into two groups: The DVT group (N = 20) and the non-DVT group (N = 60), based on whether DVT occurred within 3 days of catheterization. Single-factor and multivariate logistic regression analyses were conducted using SPSS 27.0 software. Non-DVT patients were randomly divided into a study group and a control group using a random number table method, with 30 cases in each group. The control group received routine nursing measures, while the study group received nursing measures based on NIRS for detecting DVT risk in lower limb crSaO2 in children with indwelling catheters. The blood coagulation indexes [thrombin time (TT); activated partial thromboplastin time (APTT); fibrinogen (FIB); D-dimer (D-D)] of the two groups were compared at 3 days (T0), 7 days (T1), and 14 days (T2) after catheterization. The symptoms related to lower limb DVT during the study period were also recorded. Results: Multivariate logistic analysis showed that the independent risk factors for DVT in critically ill children with deep vein catheterization were bed rest time ≥ 3 days (OR = 2.963, 95% CI = 1.282 - 6.852), limb immobilization (OR = 2.843, 95% CI = 1.209 - 6.685), FIB (OR=57.765, 95% CI = 11.308 - 295.094), and D-D (OR = 3.415, 95% CI = 1.396 - 8.359) (P &lt; 0.05). Overall, there were statistical differences in the TT, APTT, FIB, and D-D indicators, as well as time and interaction among the study group's children (P &lt; 0.05). Before catheterization, there was no statistically significant difference in TT, APTT, FIB, and D-D between the two groups of children (P &gt; 0.05). Compared between groups, the TT, APTT, FIB, and D-D at T1 and T2 in the study group were significantly lower than those in the control group, while lower limb crSaO2 was significantly higher than in the control group (P &lt; 0.05). Intra-group comparison showed statistically significant differences in TT, APTT, FIB, and D-D between the two groups of children (P &lt; 0.05). The incidence of DVT-related symptoms in the study group was significantly lower than in the control group (P &lt; 0.05). Conclusions: After deep vein catheterization, critically ill children are prone to be affected by extended bed rest and limb immobilization, resulting in the passive activation of the fibrinolytic system. Moreover, multidisciplinary team collaborative nursing measures based on NIRS technology can effectively improve coagulation function and lower limb oxygen saturation, thereby avoiding the occurrence of DVT-related symptoms.

DIFFERENCES IN PROTEASE ACTIVATED RECEPTOR-1 AND THROMBINE LEVELS IN PREECLAMPSIA AND NORMAL PREGNANCY

Introduction: Preeclampsia is a condition caused by alterations in endothelial function during pregnancy. Changes in endothelial function result in an increase in coagulation and microvascular fibrin accumulation, which results in impaired placental perfusion. Thrombin, which converts fibrin to fibrinogen, as well as platelet activity, the fibrinolytic system, and anticoagulants, are all procoagulant circumstances in preeclampsia. Thrombin contributes to the pathogenesis of preeclampsia by increasing the expression of sFlt-1 thereby providing an antiangiogenic response. Protease Activated Receptor-1 (PAR-1) is a mediator of thrombin for coagulation and inflammation in preeclampsia. Inhibition of Protease Activated Receptor-1 expression in trophoblasts can enhance placental angiogenesis and vascular remodeling. Recently, only few studies have assessed the levels of Protease Activated Receptor -7 and thrombin in preeclampsia.Objective: To determine the difference in levels of Protease Activated Receptor-1 and thrombin in preeclampsia and normal pregnancyMethods: This study is observational with a cross-sectional comparative study design. Sampling was conducted from March 2020 to March 2021. A total of 66 patients were investigated, with 33 samples of preeclampsia and 33 samples of normal pregnancy. The independent sample T-test was used for statistical analysis.Results: The mean levels of Protease Activated Receptor-1 in the preeclampsia group were higher at 28.56 ± 7.68 ng/mL while normal pregnancy was 21.67 ± 6.92 ng/mL. The results of statistical tests showed that there was a significant difference in levels of Protease Activated Receptor-1 between the preeclampsia and normal pregnancy groups (p&lt;0.05). The mean thrombin level in the preeclampsia group was higher at 72.23 ± 7.99 ng/mL, while in normal pregnancy it was 63.70 ± 8.92 ng/mL. The difference in thrombin levels between the preeclampsia and normal pregnancy groups was statistically significant (p&lt;0.05).Conclusion: Preeclampsia was associated with greater levels of Protease Activated Receptor-1 and thrombin than normal pregnancy. There was a significant difference in the mean levels of Protease Activated Receptor-1 and thrombin between preeclampsia and normal pregnancy.Keywords: Thrombin, Protease Activated Receptor-1(PAR-1), Preeclampsia

ADHESIVE DISEASE OF THE ABDOMINAL CAVITY: ETIOMORPHOPATHOGENESIS, CLINIC, DIAGNOSIS, TREATMENT AND PREVENTION AT THE PRESENT STAGE

In surgery of the gastrointestinal tract, peritoneal adhesions are detected in 80-90% of cases, including open surgical interventions, abdominal adhesions occur in 70-90% of patients, with laparoscopic – in 24-35% of patients. The number of deaths in adhesive disease ranges from 14 to 52%, and in patients with concomitant pathology, whose age exceeds sixty years, reaches 68%. The main etiological factors of the formation of adhesions are mechanical, chemical, physical and infectious effects. The pathogenesis of the formation of adhesions includes three processes: inhibition of fibrinolytic and extracellular matrix degradation systems; inflammatory reaction with cytokine production, mainly TGF-β1; tissue hypoxia as a result of interruption of blood supply to mesothelial cells and submesothelial fibroblasts. The clinical picture of SBBP, as a rule, is characterized by dyspeptic disorders in the early stages, and is accompanied by symptoms of intestinal obstruction in the advanced ones. The "gold standard". Treatment of adhesive disease can be carried out using conservative therapy or surgical intervention. To date, prevention is the most preferred method to prevent the consequences of the development of adhesive disease. Despite the improvement of surgical techniques, the development of new approaches to treatment and diagnosis, adhesions remain an inevitable consequence of intra-abdominal operations. Understanding the pathogenesis of the formation of the adhesive process and adhesion, the possibility of their transformation, especially at the cellular and molecular level, can help in the further development of more effective methods of treatment and prevention of SBP.

In vivo and in silico studies of membrane-stabilizing and clot lysis activities of Trachyspermum ammi

Trachyspermum ammi contains many important phytochemicals, including thymol, which is used to treat many diseases. This study aimed to check the membrane-stabilizing and clot-lysis capacity of the ethanol extract of T. ammi (ETA) and its main chemical component, thymol (THY). For this, we used hypotonic solution-induced erythrocyte lysing and egg-albumin protein denaturation for membrane-stabilizing capacity and human blood clot lysis for anti-atherombosis capacity. Further, we observed in silico study of THY and ntric oxide synthase (NOSs) isoform. The stabilizing effects of ETA and THY on the membrane changed depending on the concentration. At the highest concentration (160 μg/mL), they stopped the most bleeding, while the standard drug, acetylsalicylic acid, stopped less bleeding at the same concentration. In the erythrocyte lysing test, ETA showed 102.60 ± 0.01 and 75.83 ± 1.44% membrane protection. In the egg-albumin denaturation test, they showed 27.62 ± 0.02% and 56.71 ± 0.02% membrane protection, respectively. In our clot lysis experiment, ETA and THY also showed significant and concentration-dependent clot lysis capacity. The IC50 and EC50 values of THY were lower than ETA when it came to keeping the membrane stable and breaking up clots, respectively. In our in silico investigation, THY and NOSs showed significant binding interaction. Taken together, ETA and its major component, THY, exhibited potent membrane stabilizing activity and clot lysis activity in in vitro studies. Further studies are required to elucidate its other active principles and their biological effects, along with possible mechanisms.

Pharmacological mechanisms by which baicalin ameliorates cardiovascular disease.

Baicalin is a flavonoid glycoside obtained from the dried root of Scutellaria baicalensis Georgi, which belongs to the Labiatae family. Accumulating evidence indicates that baicalin has favorable therapeutic effects on cardiovascular diseases. Previous studies have revealed the therapeutic effects of baicalin on atherosclerosis, myocardial ischemia/reperfusion injury, hypertension, and heart failure through anti-inflammatory, antioxidant, and lipid metabolism mechanisms. In recent years, some new ideas related to baicalin in ferroptosis, coagulation and fibrinolytic systems have been proposed, and new progress has been made in understanding the mechanism by which baicalin protects cardiomyocytes. However, many relevant underlying mechanisms remain unexplained, and much experimental data is lacking. Therefore, further research is needed to determine these mechanisms. In this review, we summarize the mechanisms of baicalin, which include its anti-inflammatory and antioxidant effects; inhibition of endothelial cell apoptosis; modulation of innate immunity; suppression of vascular smooth muscle cells proliferation, migration, and contraction; regulation of coagulation and fibrinolytic systems; inhibition of myocardial hypertrophy; prevention of myocardial fibrosis; and anti-apoptotic effects on cardiomyocytes.

Clot lysis time and thrombin generation in patients undergoing transcatheter aortic valve implantation.

Aortic valve stenosis (AS) is the most prevalent valvular heart disease and is associated with a significant increase in mortality. AS has been shown to be linked with numerous coagulation system abnormalities, including increased fibrin deposition on the stenotic aortic valves. Transcatheter aortic valve implantation (TAVI) is the primary treatment method for patients at high surgical risk. The aim of the study was to assess the impact of treating severe AS with TAVI on thrombin generation and clot lysis time (CLT). We studied 135 symptomatic AS patients recommended for TAVI by the local Heart Team. All measurements were performed before and 5-7 days after TAVI. Alongside clinical assessment and echocardiographic analysis, we assessed clot lysis time (CLT) and thrombin generation parameters, including lag time, peak thrombin generation, time to peak thrombin generation (ttPeak), and endogenous thrombin potential (ETP). 70 patients were included in the final analysis. After TAVI, there was a significant 9% reduction in CLT despite a 12% increase in fibrinogen concentration. We observed significant increase in lag time and ttPeak (20% and 12%, respectively), and 13% decrease in peak thrombin concentration compared to pre-procedural levels. Multivariable linear regression analysis demonstrated that baseline CLT and C-reactive protein (CRP) levels were independent predictors of significant reduction in mean aortic gradient, defined as TAVI procedure success. CLT and peak thrombin concentration decreased, while Lag time and ttPeak increased significantly after TAVI. Multivariable linear regression analysis demonstrated CLT and CRP levels as independent predictors of achieving a reduction in mean aortic gradient, defining TAVI procedure success.

Emerging Adjuvant Thrombolytic Therapies for Acute Ischemic Stroke Reperfusion.

Thrombolytic therapies for acute ischemic stroke are widely available but only result in recanalization early enough, to be therapeutically useful, in 10% to 30% of cases. This large gap in treatment effectiveness could be filled by novel therapies that can increase the effectiveness of thrombus clearance without significantly increasing the risk of harm. This focused update will describe the current state of emerging adjuvant treatments for acute ischemic stroke reperfusion. We focus on new treatments that are designed to (1) target different components that make up a stroke thrombus, (2) enhance endogenous fibrinolytic systems, (3) reduce stagnant blood flow, and (4) improve recanalization of distal thrombi and postendovascular thrombectomy.

Moonlighting on the Fasciola hepatica tegument: Enolase, a glycolytic enzyme, interacts with the extracellular matrix and fibrinolytic system of the host.

Enolase is a 47 kDa enzyme that functions within the glycolysis and gluconeogenesis pathways involved in the reversible conversion of D-2-phosphoglycerate (2PGA) to phosphoenolpyruvate (PEP). However, in the context of host-pathogen interactions, enolase from different species of parasites, fungi and bacteria have been shown to contribute to adhesion processes by binding to proteins of the host extracellular matrix (ECM), such as fibronectin (FN) or laminin (LM). In addition, enolase is a plasminogen (PLG)-binding protein and induces its activation to plasmin, the main protease of the host fibrinolytic system. These secondary 'moonlighting' functions of enolase are suggested to facilitate pathogen migration through host tissues. This study aims to uncover the moonlighting role of enolase from the parasite Fasciola hepatica, shedding light on its relevance to host-parasite interactions in fasciolosis, a global zoonotic disease of increasing concern. A purified recombinant form of F. hepatica enolase (rFhENO), functioning as an active homodimeric glycolytic enzyme of ~94 kDa, was successfully obtained, fulfilling its canonical role. Immunoblotting studies on adult worm extracts showed that the enzyme is present in the tegument and the excretory/secretory products of the parasite, which supports its key role at the host-parasite interface. Confocal immunolocalisation studies of the protein in newly excysted juveniles and adult worms also localised its expression within the parasite tegument. Finally, we showed by ELISA that rFhENO can act as a parasitic adhesin by binding host LM, but not FN. rFhENO also binds PLG and enhances its conversion to plasmin in the presence of the tissue-type and urokinase-type PLG activators (t-PA and u-PA). This moonlighting adhesion-like function of the glycolytic protein enolase could contribute to the mechanisms by which F. hepatica efficiently invades and migrates within its host and encourages further research efforts that are designed to impede this function by vaccination or drug design.

Fibrinogen and plasma clot properties are associated with apolipoprotein B and apolipoprotein B-containing lipoproteins in Africans

Fibrinogen and plasma clot properties are associated with apolipoprotein B and apolipoprotein B-containing lipoproteins in Africans

Fibrinolytic Platelet Decoys Reduce Cancer Metastasis by Dissociating Circulating Tumor Cell Clusters.

During metastasis, circulating tumor cells (CTCs) can travel in the bloodstream as individual cells or clusters, associated with fibrin and platelets. Clusters have a higher metastatic potential due to their increased ability to withstand shear stress and arrest in small vessels. Moreover, CTC-platelet interaction protects CTCs from shear stress and immune detection. The objective of this project is to develop a fibrinolytic platelet system to leverage platelet-CTC interactions and dissociate CTC clusters. For this approach, tissue plasminogen activator (tPA) is loaded onto two modified platelet systems: platelet Decoys and lyophilized platelets. The activities of the systems are characterized using a Förster Resonance Energy Transfer-based assay and an angiogenic assay. Furthermore, the ability of the system to dissociate cancer cell clusters in vitro is assessed using light transmission aggregometry. The data demonstrates that the fibrinolytic platelets can maintain tPA activity, interact with CTCs, and dissociate cancer cell clusters. Finally, fibrinolytic platelets are assessed in vivo, demonstrating a decreased tumor load and increased survival with tPA-Decoy treatment, which is selected as the optimal treatment based on favorable in vitro results and in vivo trials. Therefore, this fibrinolytic platelet approach is a promising method for leveraging platelet-CTC interactions to disperse CTC clusters and reduce metastasis.

Levels of markers of coagulation and fibrinolysis systems in patients with pulmonary tuberculosis with concomitant diabetes mellitus after COVID-19

Background. It is known that COVID-19 can be followed by a shift in the hemostatic system towards hypercoagulation, which is more pronounced in the presence of diabetes mellitus (DM). Tuberculosis process is often accompanied with hypercoagulation syndrome. Of great interest is the study of the state of hemostatic systems in patients with pulmonary tuberculosis (TB) with concomitant DM who have had COVID-19.The aim. To study the relationship between the state of the hemostatic and fibrinolysis systems and moderate and severe COVID-19 in patients with pulmonary tuberculosis and diabetes mellitus.Methods. 32 patients with TB and DM were divided into two groups. Group 1 included 16 patients with TB and DM who have previously had COVID-19 (TB-DM-COVID). Group 2 included 16 patients with TB and DM who did not have COVID-19 (TB-DM).Results. It was found that TB-DM-COVID patients were more likely to develop a hypercoagulable shift compared to TB-DM patients. This was evidenced by a more frequent shortening of such indicator as activated partial thromboplastin time (43.7 % and 25.0 % of cases, respectively; χ2 = 7.22; p = 0.01), an increase in fibrinogen levels (43.7 % and 25.0%, respectively; χ2 = 7.22; p = 0.01) and D-dimer (43.7 % and 18.7 %, respectively; χ2 = 14.74; p = 0.0001). These changes were closely associated with the systemic inflammatory response, as strong and positive correlations were found between fibrinogen and C-reactive protein levels (r = 0.420; p = 0.01), and erythrocyte sedimentation rate (r = 0.433; p = 0.01) in TB-DM-COVID patients.Conclusion. In patients with pulmonary tuberculosis and diabetes mellitus after moderate and severe COVID-19, compared to patients who have not had COV ID-19, a hypercoagulable shift associated with the development of more pronounced systemic inflammation develops more often.