Fibrinolytic Research Articles

Isolation and Characterization of Alkaline Proteases Producing Indigenous Bacillus sp. as a Source of Thrombolytic and Fibrinolytic Agents

Fibrinolytic enzymes derived from bacteria can be effective as a potent, safe and cost-effective thrombolytic agent for the prevention and treatment of cardiovascular diseases. In this study, microorganisms were isolated from various soil samples and screened for their protease activity and protein concentration. Isolates with higher protease activity were subsequently tested for their thrombolytic and fibrinolytic activity. Of a total 37 isolates, 7 exhibited significant zone in SMA media (zone ratio above 1.3). Of these, 8 isolates showed increased protease activity with values ranging from 120 to 199 U/ml. The highest protease activity was observed for the isolate GST12 obtained from mung bean (green gram) soil. The freeze-dried concentrated crude enzymes were screened by in vitro clot lysis method. Six of these eight concentrated enzymes demonstrated clot lysis above 20%, whereas, enzymes from GST12 and GST21 exhibited the highest clot lysis activity (about 27%), which is comparable with standard drug streptokinase. These two enzymes are also strongly associated with fibrinolytic activity. Our findings identify 8 isolates having potential thrombolytic and fibrinolytic activities that belong to Gram-positive Bacillus. Dhaka Univ. J. Pharm. Sci. 22(2): 125-135, 2023 (December)

A Noninvasive Circulating Signature of Combined Right Ventricular Pressure and Volume Overload in Tetralogy of Fallot/Pulmonary Atresia/Major Aortopulmonary Collateral Arteries.

Background: Despite surgical advances, children with tetralogy of Fallot/pulmonary atresia/major aortopulmonary collaterals (TOF/PA/MAPCAs) are subject to chronic right ventricular (RV) pressure and volume overload. Current diagnostic tools do not identify adverse myocardial remodeling and cannot predict progression to RV failure. We sought to identify a noninvasive, circulating signature of the systemic response to right heart stress to follow disease progression. Methods: Longitudinal data were collected from patients with TOF/PA/MAPCAs (N = 5) at the time of (1) early RV pressure overload and (2) late RV pressure and volume overload. Plasma protein and microRNA expression were evaluated using high-throughput data-independent mass spectroscopy and Agilent miR Microarray, respectively. Results: At the time of early RV pressure overload, median patient age was 0.34 years (0.02-9.37), with systemic RV pressures, moderate-severe hypertrophy, and preserved systolic function. Late RV pressure and volume overload occurred at a median age of 4.08 years (1.51-10.83), with moderate RV hypertrophy and dilation, and low normal RV function; 277 proteins were significantly dysregulated (log2FC ≥0.6/≤-0.6, FDR≤0.05), predicting downregulation in lipid transport (apolipoproteins), fibrinolytic system, and extracellular matrix structural proteins (talin 1, profilin 1); and upregulation in the respiratory burst. Increasing RV size and decreasing RV function correlated with decreasing structural protein expression. Similarly, miR expression predicted downregulation of extracellular matrix-receptor interactions and upregulation in collagen synthesis. Conclusion: To our knowledge, we show for the first time a noninvasive protein and miR signature reflecting the systemic response to adverse RV myocardial remodeling in TOF/PA/MAPCAs which could be used to follow disease progression.

Indicators of hemostasis and fibrinolysis systems and systemic inflammatory response in patients with newly diagnosed sarcoidosis after COVID-19

Objective. To study the indicators of hemostasis and fibrinolysis systems, as well as the systemic inflammatory response in patients with newly diagnosed sarcoidosis after infection caused by SARS-CoV-2. Material and methods. A prospective analysis of the indicators of hemostasis and fibrinolysis systems, as well as the systemic inflammatory response of 20 patients with sarcoidosis, after COVID-19, who were on inpatient treatment in Central TB Research Institute. The patients were divided into 2 groups. The first group included 10 patients with sarcoidosis, who had suffered COVID-19 (the main group). The comparison group consisted of 10 patients with sarcoidosis without a history of COVID-19. Results. It was found, that in patients with newly diagnosed sarcoidosis, who underwent COVID-19, there is a hypercoagulation shift in the hemostasis and fibrinolysis system. This was evidenced by a significant shortening of the indicators of APTT, PT and an increase in the level of D-dimer compared to both healthy and patients, who had not undergone COVID-19. In addition, laboratory manifestations of systemic inflammation were most pronounced in patients with sarcoidosis who underwent COVID-19. Probably, changes in the hemostasis and fibrinolysis system occurred within the framework of a systemic inflammatory response.

CLOT LYSIS AND MEMBRANE PROTECTION POTENTIALS OF CHEILANTHES TENUIFOLIA METHANOLIC LEAF EXTRACT

Cheilanthes tenuifolia is a little perennial fern that falls within the taxonomic classification of the Pteridaceae botanical family. The plant has a diverse array of phytochemical substances, including alkaloids, phenolic compounds, flavonoids, saponins, steroids, and triterpenoids, which have demonstrated promising medicinal properties. This study aimed to evaluate the in-vitro membrane-stabilizing and clot lysis activities of a methanol leaf extract of C. tenuifolia (MCT). For this, we performed hypotonic solution-induced erythrocyte lysing and human blood clot lysis methods to check the membrane stabilizing and clot lysis capacities of MCT using acetylsalicylic acid and streptokinase as standards, respectively. Additionally, we also checked its phytochemical groups. The results of a preliminary phytochemical screening indicate the presence of alkaloids, glycosides, tannins, flavonoids, and saponins in the plant. MCT inhibited hemolysis in a concentration-dependent manner and inhibited 78.93 ± 0.01% hemolysis (IC50 = 46 ± 2.11 µg/ml) at the higher concentration (160 μg/ml), whereas the standard drug, acetylsalicylic acid (IC50 = 64.10 ± 2.08 µg/ml) inhibited 97.71 ± 0.01% at the same concentration. It also exhibited clot lysis in a concentration-dependent manner, where the maximum percentage of clot lysis was observed at 160 μg/100 ml where the IC50 value was 198.41 ± 1.87 µg. The standard drug streptokinase showed 77.51 ± 0.01% clot lysis. C. tenuifolia possesses various important secondary metabolites and shows membrane stabilizing and clot lysis capacity. Further studies are required to elucidate its active principles and their biological effects.

Validation of the time to attain maximal clot amplitude after reaching maximal clot formation velocity parameter as a measure of fibrinolysis using rotational thromboelastometry and its application in the assessment of fibrinolytic resistance in septic patients: a prospective observational study: communication from the ISTH SSC Subcommittee on Fibrinolysis

BackgroundIn sepsis, fibrinolysis resistance correlates with worse outcomes. Practically, rotational thromboelastometry (ROTEM) is used to report residual clot amplitude relative to maximum amplitude at specified times after clot formation clot lysis indices (CLIs). However, healthy individuals can exhibit similar CLIs, thus making it challenging to solely diagnose the low fibrinolytic state. Furthermore, CLI does not include the kinetics of clot formation, which can affect overall fibrinolysis. Therefore, a more nuanced analysis, such as time to attain maximal clot amplitude after reaching maximal clot formation velocity (t-AUCi), is needed to better identify fibrinolysis resistance in sepsis. ObjectivesTo evaluate the correlation between the degree of fibrinolytic activation and t-AUCi in healthy or septic individuals. MethodsWhole blood (n = 60) from septic or healthy donors was analyzed using tissue factor–activated (EXTEM) and nonactivated (NATEM) ROTEM assays. Lysis was initiated with tissue-type plasminogen activator, and CLI and t-AUCi were calculated. Standard coagulation tests and plasma fibrinolysis markers (D-dimer, plasmin-α2-antiplasmin complex, plasminogen activator inhibitor type 1, and plasminogen) were also measured. Resultst-AUCi values decreased with increasing fibrinolytic activity and correlated positively with CLI for different degrees of clot lysis both in EXTEM and NATEM. t-AUCi cutoff value of 1962.0 seconds in EXTEM predicted low fibrinolytic activity with 81.8% sensitivity and 83.7% specificity. In addition, t-AUCi is not influenced by clot retraction. ConclusionWhole-blood point-of-care ROTEM analyses with t-AUCi offers a more rapid and parametric evaluation of fibrinolytic potential compared with CLI, which can be used for a more rapid and accurate diagnosis of fibrinolysis resistance in sepsis.

Novel Insights into the Link Between Myeloperoxidase Modified LDL, LOX-1, and Neuroserpin in Stroke.

Cardiovascular disease that is caused by atherosclerosis is the leading cause of death worldwide. Atherosclerosis is primarily triggered by endothelial dysfunction and the accumulation of modified low-density lipoprotein (LDL) particles in the subendothelial space of blood vessels. Early reports have associated oxidized LDL with altered fibrinolysis and atherogenesis. It has been suggested that myeloperoxidase oxidized LDL (Mox-LDL) is involved in atherosclerosis because of its significant pathophysiological role in the modification of LDL in vivo. It has been equally demonstrated that Mox-LDL binds to the lectin-like oxidized low-density lipoprotein receptor-1 (lox-1) scavenger receptor which leads to the upregulation of inflammatory mediators in endothelial cells and the progression of cardiovascular disease. It has been also shown that neuroserpin, a member of the serine proteinase inhibitor (serpin) superfamily, has an important role at the level of fibrinolysis in the nervous tissue. Since little is known about the effects of Mox-LDL on endothelial cell fibrinolytic activity and the involvement of lox-1 in this process, our study aimed at evaluating the in vitro effects of Mox-LDL on neuroserpin release from human aortic endothelial cells (HAECs) and the role of lox-1 scavenger receptor in this context by relying on lox-1 gene silencing in HAECs, culturing the cells in the presence of Mox-LDL, measuring their neuroserpin expression and release by quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA), respectively, and assessing their fibrinolytic activity using the Euglobulin Clot Lysis Time (ECLT) method. Our data show that Mox-LDL decreases endothelial cell fibrinolytic capacity by upregulating neuroserpin in lox-1 knockdown cells. Lox-1 protects the endothelial cells from a Mox-LDL-induced decrease in pro-fibrinolytic capacity, which has important consequences in the context of stroke.

Highly selective fluorometric detection of streptokinase via fibrinolytic release of photoluminescent carbon dots integrated into fibrin clot network

This work pioneers a selective fluorometric assay for the fibrinolytic agent streptokinase by strategically coupling its thrombolytic action to fluorescence signaling using carbon dots integrated into fibrin clot network. Streptokinase triggers clot lysis and caused release of photoluminescent carbon dots integrated within the fibrin network in a concentration-dependent manner. Transmission electron microscope (TEM) imaging verifies distinct aggregation of the carbon dots upon addition to fibrin. The carbon dots are derived via an eco-friendly hydrothermal approach using cucumber peel as the source of carbon. Structural and optical characterization of the carbon dots were performed using different techniques including X-ray diffraction, TEM, Fourier-transform infrared spectroscopy, and ultraviolet–visible absorption and fluorescence spectroscopies. All experimental parameters influencing the formation of the fibrin clot and factors governing the release of carbon dots upon the addition of streptokinase have been fully optimized. This fluorescence assay offers high selectivity for streptokinase over potential interferences in addition to excellent sensitivity with a 0.017 μg/mL limit of detection and a wide linear range (0.05–500 μg/mL). Successful application to pharmaceutical formulations is demonstrated with excellent recovery and precision. By ingeniously linking streptokinase activity to carbon dot fluorescence, this novel assay provides a rapid, simple and eco-friendly method for selective streptokinase detection.

Developmental adcyap1b loss leads to hemorrhage, disrupted hemostasis, and a blood coagulation cascade in zebrafish

BackgroundPituitary adenylate cyclase–activating polypeptide is a neuropeptide with diverse roles in biological processes. Its involvement in the blood coagulation cascade is unclear. ObjectivesThis study unraveled adcyap1b’s role in blood coagulation using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 in zebrafish. Effects were validated via adcyap1b knockdown. Gene expression changes in adcyap1b mutants were explored, linking them to clotting disorders. An analysis of proca gene splicing illuminated its role in adcyap1b-related anticoagulation deficiencies. MethodsZebrafish were genetically modified using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 to induce adcyap1b knockout. Morpholino-mediated gene knockdown was employed for validation. Expression levels of coagulation factors, anticoagulant proteins, and fibrinolytic system genes were assessed in adcyap1b mutant zebrafish. Alternative splicing of proca gene was analyzed. ResultsAdcyap1b mutant zebrafish exhibited severe hemorrhage, clotting disorders, and disrupted blood coagulation. Morpholino-mediated knockdown replicated observed phenotypes. Downregulation in transcripts related to coagulation factors V and IX, anticoagulation protein C, and plasminogen was observed. Abnormal alternative splicing of the proca gene was identified, providing a mechanistic explanation for anticoagulation system deficiencies. ConclusionAdcyap1b plays a crucial role in maintaining zebrafish blood coagulation and hemostasis. Its influence extends to the regulation of procoagulant and anticoagulant pathways, with abnormal alternative splicing contributing to observed deficiencies. These findings unveil a novel aspect of adcyap1b function, offering potential insights into similar processes in mammalian systems.

Low factor XIII levels and altered fibrinolysis in patients with multiple myeloma

BackgroundAcquired factor FXIII (FXIII) deficiency can be immune- or non-immune mediated and may cause severe bleeding symptoms. The incidence of acquired FXIII deficiency and its etiology in patients with multiple myeloma (MM) are poorly understood. ObjectivesTo assess FXIII levels and the balance of fibrinolysis in newly diagnosed, untreated MM and monoclonal gammopathy of undetermined significance (MGUS) patients. MethodsFXIII activity, mixing studies, FXIII-A2B2 antigen, total FXIII-B antigen were measured in platelet-poor plasma from 17 untreated MM patients, 33 untreated MGUS patients, and 30 age and sex-matched healthy controls. Besides routine laboratory measurements, the balance of coagulation and fibrinolysis was evaluated using quantitative fibrin monomer (FM) test, thrombin-antithrombin assay, α2-antiplasmin activity, plasmin-α2-antiplasmin (PAP) complex, D-dimer, plasmin generation assay, clot lysis assay, and ClotPro-TPA test. ResultsFXIII-A2B2 levels were significantly lower in MM patients compared to controls [median (IQR):14.6 (11.2–19.4) vs. 21.8 (17.1–26.4) mg/L, p = 0.0015], whereas total FXIII-B did not differ between groups. Decrease in FXIII activity was parallel to the decrease in FXIII-A2B2. An immune-mediated inhibitory mechanism was ruled out. Free/total FXIII-B was significantly higher in MM patients compared to MGUS and healthy controls, suggesting an etiology of FXIII-A consumption. In MM and MGUS patients, FM, D-dimer, and PAP complex were significantly elevated compared to controls, indicating hypercoagulability and ongoing fibrinolysis. ConclusionsLow FXIII levels due to consumption were observed in MM patients at diagnosis. Hypercoagulability and ongoing fibrinolysis were detected in MM and MGUS, indicating that a disturbed hemostasis balance is already present in the latter benign condition.

TAFI phenomenon and its clinical significance

Thrombin activatable fi brinolysis inhibitor (TAFI) is one of the key proteins of fi brinolysis and infl ammation systems. Based on literature data, information is presented on the biological eff ects of TAFI, the mechanisms of its activation, gene polymorphism that determines the duration of its action and activity. Nosological forms are listed, where TAFI is important in the pathogenesis of the development of diseases, its role in experimental and laboratory studies, and the possibility of pharmacological infl uence on the activity of this protein.

Involvement of plasminogen activator inhibitor-1 in p300/p53-mediated age-related atrial fibrosis.

Plasminogen activator inhibitor-1 (PAI-1), a key regulator of the fibrinolytic system, is also intimately involved in the fibrosis. Although PAI-1 may be involved in the occurrence of atrial fibrillation (AF) and thrombosis in the elderly, but whether it participated in aging-related atrial fibrosis and the detailed mechanism is still unclear. We compared the transcriptomics data of young (passage 4) versus senescent (passage 14) human atrial fibroblasts and found that PAI-1 was closely related to aging-related fibrosis. Aged mice and senescent human and mouse atrial fibroblasts underwent electrophysiological and biochemical studies. We found that p300, p53, and PAI-1 protein expressions were increased in the atrial tissue of aged mice and senescent human and mouse atrial fibroblasts. Curcumin or C646 (p300 inhibitor), or p300 knockdown inhibited the expression of PAI-1 contributing to reduced atrial fibroblasts senescence, atrial fibrosis, and the AF inducibility. Furthermore, p53 knockdown decreased the protein expression of PAI-1 and p21 in senescent human and mouse atrial fibroblasts. Our results suggest that p300/p53/PAI-1 signaling pathway participates in the mechanism of atrial fibrosis induced by aging, which provides new sights into the treatment of elderly AF.

Evaluation of the antioxidant, thrombolytic, and antimicrobial effects of Corchorus aestuans L. leaf extracts: an in-vitro and in-silico study

Corchorus aestuans L. (Malvaceae), a little annual plant, grows in wastelands at low and medium altitudes. It has traditionally been used as an antihistaminic, anti-inflammatory, and antibacterial for chronic cystitis, gonorrhea, and dysuria. Our research aimed to investigate the antioxidant, antimicrobial, and thrombolytic potential of several solvent extracts. Three different solvents were used in the extraction process: methanol, hydro-methanol, and water. Using established qualitative methodologies, phytochemical screening was carried out. Folin-Ciocalteu and aluminum chloride techniques were used to determine the total phenol and flavonoid contents. The antioxidant activity (total antioxidant capacity, DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging, and Fe3+ reducing power) was evaluated using spectrophotometric methods. Besides, clot lysis and disc-diffusion methods were used to assess thrombolytic and antibacterial activity. The GC-MS analysis was also performed to discover prominent phytochemicals. Phytochemical screening revealed the presence of glycosides, alkaloids, flavonoids, phenols, tannins, etc. Among all the extracts, hydro-methanol extract (MACA) possesses the highest total phenol and flavonoid contents (778.2±20.82 mg GAE/g dry extract and 535.44±20.95 mg QE/g dry extract respectively). Similarly, MACA showed the highest total antioxidant capacity (664.31±16.84mg/g equivalent to AA), DPPH scavenging activity (IC50: 26.30±2.82 μg/mL), and ferric reducing capacity (EC50: 18.58±2.37 μg/mL). Moreover, among the three extracts, MACA showed the highest percentages of clot lysis (40.49 ±1.97 %) and an antibacterial effect against B. subtilis. GC-MS revealed 17 phytoconstituents, whereas (1s,15s) bicyclo [13.1.0] hexadecan-2-one had the greatest binding score with 6NGJ (-7.9 kcal/mol), and 1A5H (-6.9 kcal/mol) proteins. The ADMET studies showed they are non-carcinogenic, and well-absorbed, while PASS prediction and drug-likeness data showed antioxidant, anti-inflammatory, and good oral bioavailability. Therefore, further research will potentiate to discover compounds with antioxidant, thrombolytic, and antibacterial properties, with their mechanism of action.

Specific inhibition on PAI-1 reduces the dose of Alteplase for ischemic stroke treatment

Administration of recombinant tPA (rtPA, or trade name Alteplase®) is an FDA-approved therapy for acute ischemic stroke (AIS), but poses the risk of hemorrhagic complications. Recombinant tPA can be rapidly inactivated by the endogenous inhibitor, plasminogen activator inhibitor 1 (PAI-1). In this work, we study a novel treatment approach that combines a PAI-1 inhibitor, PAItrap4, with a reduced dose of rtPA to address the hemorrhagic concern of rtPA. PAItrap4 is a highly specific and very potent protein-based inhibitor of PAI-1, comprising of a variant of uPA serine protease domain, human serum albumin, and a cyclic RGD peptide. PAItrap4 efficiently targets and inhibits PAI-1 on activated platelets, and also possesses a long half-life in vivo. Our results demonstrate that PAItrap4 effectively counteracts the inhibitory effects of PAI-1 on rtPA, preserving rtPA activity based on amidolytic and clot lysis assays. In an in vivo murine stroke model, PAItrap4, together with low-dose rtPA, enhances the blood perfusion in the stroke-affected areas, reduces infarct size, and promotes neurological recovery in mice. Importantly, such treatment does not increase the amount of cerebral hemorrhage, thus reducing the risk of cerebral hemorrhage. In addition, PAItrap4 does not compromise the normal blood coagulation function in mice, demonstrating its safety as a therapeutic agent. These findings highlight this combination therapy as a promising alternative for the treatment of ischemic stroke, offering improved safety and efficacy.

Diagnostic Utility of a Combined MPO/D-Dimer Score to Distinguish Abdominal Aortic Aneurysm from Peripheral Artery Disease.

Abdominal aortic aneurysm (AAA) and peripheral artery disease (PAD) share pathophysiological mechanisms including the activation of the fibrinolytic and innate immune system, which explains the analysis of D-dimer and myeloperoxidase (MPO) in both conditions. This study evaluates the diagnostic marker potential of both variables separately and as a combined MPO/D-dimer score for identifying patients with AAA versus healthy individuals or patients with PAD. Plasma levels of MPO and D-dimer were increased in PAD and AAA compared to healthy controls (median for MPO: 13.63 ng/mL [AAA] vs. 11.74 ng/mL [PAD] vs. 9.16 ng/mL [healthy], D-dimer: 1.27 μg/mL [AAA] vs. 0.58 μg/mL [PAD] vs. 0.38 μg/mL [healthy]). The combined MPO/D-dimer score (median 1.26 [AAA] vs. -0.19 [PAD] vs. -0.93 [healthy]) showed an improved performance in distinguishing AAA from PAD when analysed using the receiver operating characteristic curve (area under the curve) for AAA against the pooled data of healthy controls + PAD: 0.728 [MPO], 0.749 [D-dimer], 0.801 [score]. Diagnostic sensitivity and specificity ranged at 82.9% and 70.2% (for score cut-off = 0). These findings were confirmed for a separate collective of AAA patients with 35% simultaneous PAD. Thus, evaluating MPO together with D-dimer in a simple score may be useful for diagnostic detection and the distinction of AAA from athero-occlusive diseases like PAD.

Effects of proprotein convertase subtilisin-kexin type 9 inhibitors on inflammatory and hemostatic parameters in post myocardial infarction patients

Despite progress in treatment, elevated levels of low-density lipoprotein cholesterol (LDL-C) and lipoprotein (a) (Lp(a)), represent a significant part of the residual risk. Both are associated with inflammation and the coagulation fibrinolytic system. The purpose of our research was to evaluate the effect of proprotein convertase subtilisin/kexin type 9 inhibitors (PCSK9i) on lipid parameters and indicators of inflammation, coagulation and fibrinolysis. We included 100 post myocardial infarction (MI) patients with insufficiently controlled LDL-C values despite the maximum dose of statin, and highly elevated Lp(a). Patients received alirocumab or evolocumab (150 mg sc or 140 mg sc every two weeks, respectively), or placebo for 6 months.In patients receiving PCSK9i, a significant decrease in total cholesterol (TC), LDL-C, triglycerides (TG) and Lp(a), and an increase in high density lipoprotein cholesterol (p < 0.001 for all) was found. Before treatment, the concentrations of TC, LDL-C and TG correlated with the concentrations of thrombin activatable fibrinolysis inhibitor (r = 0.41, p < 0.001; r = 0.353, p < 0.001; r = 0.311, p = 0.003, respectively), and plasminogen activator inhibitor-1 (r = 0.302, p = 0.007; r = 0.218, p = 0.049; r = 0.278; p = 0.013, respectively). The concentrations of TC and LDL-C correlated with overall fibrinolytic potential (r = −0.220, p = 0.034; r = −0.207, p = 0.047, respectively). The concentration of TG was related to the concentration of interleukin 6 (r = 0.290, p = 0.004) and interleukin 8 (r = 0.332, p = 0.001). No correlations between Lp(a) and inflammatory or hemostatic variables were found. No associations were found after treatment. Our results show that inflammatory cytokines and fibrinolytic parameters are related to LDL-C and not Lp(a) in post-MI patients before and with neither of them following PCSK9i treatment.The trial registration number: NCT04613167, Date of registration: November 3, 2020.

Vasoinhibin's Apoptotic, Inflammatory, and Fibrinolytic Actions Are in a Motif Different From Its Antiangiogenic HGR Motif.

Vasoinhibin, a proteolytic fragment of the hormone prolactin, inhibits blood vessel growth (angiogenesis) and permeability, stimulates the apoptosis and inflammation of endothelial cells, and promotes fibrinolysis. The antiangiogenic and antivasopermeability properties of vasoinhibin were recently traced to the HGR motif located in residues 46 to 48 (H46-G47-R48), allowing the development of potent, orally active, HGR-containing vasoinhibin analogues for therapeutic use against angiogenesis-dependent diseases. However, whether the HGR motif is also responsible for the apoptotic, inflammatory, and fibrinolytic properties of vasoinhibin has not been addressed. Here, we report that HGR-containing analogues are devoid of these properties. Instead, the incubation of human umbilical vein endothelial cells with oligopeptides containing the sequence HNLSSEM, corresponding to residues 30 to 36 of vasoinhibin, induced apoptosis, nuclear translocation of NF-κB, expression of genes encoding leukocyte adhesion molecules (VCAM1 and ICAM1) and proinflammatory cytokines (IL1B, IL6, and TNF), and adhesion of peripheral blood leukocytes. Also, intravenous or intra-articular injection of HNLSSEM-containing oligopeptides induced the expression of Vcam1, Icam1, Il1b, Il6, and Tnf in the lung, liver, kidney, eye, and joints of mice and, like vasoinhibin, these oligopeptides promoted the lysis of plasma fibrin clots by binding to plasminogen activator inhibitor-1 (PAI-1). Moreover, the inhibition of PAI-1, urokinase plasminogen activator receptor, or NF-κB prevented the apoptotic and inflammatory actions. In conclusion, the functional properties of vasoinhibin are segregated into 2 different structural determinants. Because apoptotic, inflammatory, and fibrinolytic actions may be undesirable for antiangiogenic therapy, HGR-containing vasoinhibin analogues stand as selective and safe agents for targeting pathological angiogenesis.

Evaluation of phytochemical, antibacterial, thrombolytic, anti-inflammatory, and cytotoxicity profile of Achyranthes aspera aerial part extracts

This investigation was designed and performed to compare the phytochemical profiling, activities of antibacterial, thrombolytic, anti-inflammatory, and cytotoxicity of methanol extract (ME-E) and aqueous extract (AQ-E) of aerial parts of Achyranthes aspera through in-vitro approach. Also characterize the functional groups of bioactive compounds in the ME-E through Fourier-transform infrared (FTIR) spectroscopy analysis. Interestingly, qualitative phytochemical screening proved that the ME-E contain more number of vital phytochemicals such as phenolics. saponins, tannins, alkaloids, flavonoids, cardiac glycosides, steroids, and phlobatannins than AQ-E. Similarly, the ME-E showed notable antibacterial activity as dose dependent manner against Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa at 1000 μg mL−1 concentration. ME-E also showed 75.2 ± 2% of clot lysis (thrombolytic activity) at 1000 μg mL−1 dosage and it followed by AQ-E 51.24 ± 3%. The ME-E showed moderate and AQ-E demonstrate poor anti-inflammatory activity evidenced by albumin denaturation inhibition and anti-lipoxygenase assays. Furthermore, the ME-E demonstrated a dose dependent cytotoxicity was noted against brine shrimp larvae. In support of this ME-E considerable activities, the Fourier transform infrared (FTIR) analysis confirmed that this extract contain more number peaks attributed to the stretch of various essential functional groups belongs to different bioactive compounds. Hence this ME-E of A. aspera can be considered for further in depth scientific investigations to validate their maximum biomedical potential.

Cyclical endometrial repair and regeneration: Molecular mechanisms, diseases, and therapeutic interventions.

The endometrium is a unique human tissue with an extraordinary ability to undergo a hormone-regulated cycle encompassing shedding, bleeding, scarless repair, and regeneration throughout the female reproductive cycle. The cyclical repair and regeneration of the endometrium manifest as changes in endometrial epithelialization, glandular regeneration, and vascularization. The mechanisms encompass inflammation, coagulation, and fibrinolytic system balance. However, specific conditions such as endometriosis or TCRA treatment can disrupt the process of cyclical endometrial repair and regeneration. There is uncertainty about traditional clinical treatments' efficacy and side effects, and finding new therapeutic interventions is essential. Researchers have made substantial progress in the perspective of regenerative medicine toward maintaining cyclical endometrial repair and regeneration in recent years. Such progress encompasses the integration of biomaterials, tissue-engineered scaffolds, stem cell therapies, and 3D printing. This review analyzes the mechanisms, diseases, and interventions associated with cyclical endometrial repair and regeneration. The review discusses the advantages and disadvantages of the regenerative interventions currently employed in clinical practice. Additionally, it highlights the significant advantages of regenerative medicine in this domain. Finally, we review stem cells and biologics among the available interventions in regenerative medicine, providing insights into future therapeutic strategies.

Value of novel thrombotic markers for predicting occurrence of the malignant cerebral artery infarction: a prospective clinical study

ObjectiveThis study investigated the diagnostic performance of thrombin–antithrombin complex (TAT), plasmin–α2 plasmin inhibitor complex (PIC), tissue plasminogen activator–plasminogen activator inhibitor complex (t-PAIC), and thrombomodulin (TM) in predicting the progression of massive cerebral infarction to the malignant cerebral artery infarction.MethodA total of 71 patients with massive cerebral infarction confirmed by imaging examination were divided into malignant cerebral artery infarction group (MCAI) and non-malignant cerebral artery infarction group (NMCAI) based on whether they progressed to MCAI after admission. TAT, PIC, t-PAIC, and TM were measured immediately after admission. The predictive performance was analyzed by the receiver characteristic operating curve (ROC).ResultThe median plasma concentrations of TM, PIC, TAT, and t-PAIC in the MCAI patients at admission were 10.65 IU/mL, 1.17 μg/mL, 12.25 ng/mL, and 13.85 ng/mL, respectively, which were higher than those in the NMCAI patients (9.00 IU/mL, 1.07 μg/mL, 4.60 ng/mL, and 8.70 ng/mL), and the difference was statistically significant (p = 0.045, p = 0.035, p = 0.004, and p = 0.003). Elevated plasma t-PAIC concentration was shown to be an independent risk factor for progression of massive cerebral infarction to MCAI (OR = 1.131) by multivariate logistic regression analysis. ROC curve analysis showed that t-PAIC was the best predictor of MCAI (AUC = 74.7%), with a sensitivity of 75.0% and specificity of 75.9% when t-PAIC concentration was ≥12.4 ng/mL; TAT had the highest specificity in predicting MCAI, with a specificity of 90.7% when the TAT concentration was ≥13.5 ng/mL.ConclusionThe detection of PIC, TAT, t-PAIC, and TM is a comprehensive assessment of vascular endothelial damage and activation of the coagulation and fibrinolytic systems and has predictive value for poor prognosis in patients with MCAI. The widespread use of these tests will likely greatly improve the early diagnosis rate of MCAI.

Hemorrhagic Coagulation Disorders and Ischemic Stroke: How to Reconcile Both?

Coagulation and fibrinolytic system disorders are conditions in which the blood's ability to clot is impaired, resulting in an increased risk of thrombosis or bleeding. Although these disorders are the expression of two opposing tendencies, they can often be associated with or be a consequence of each other, contributing to making the prognosis of acute cerebrovascular events more difficult. It is important to recognize those conditions that are characterized by dual alterations in the coagulation and fibrinolytic systems to reduce the prognostic impact of clinical conditions with difficult treatment and often unfortunate outcomes. Management of these individuals can be challenging, as clinicians must balance the need to prevent bleeding episodes with the potential risk of clot formation. Treatment decisions should be made on an individual basis, considering the specific bleeding disorder, its severity, and the patient's general medical condition. This review aims to deal with all those forms in which coagulation and fibrinolysis represent two sides of the same media in the correct management of patients with acute neurological syndrome. Precision medicine, personalized treatment, advanced anticoagulant strategies, and innovations in bleeding control represent future directions in the management of these complex pathologies in which stroke can be the evolution of two different acute events or be the first manifestation of an occult or unknown underlying pathology.