scholarly journals The Anti-platelet and Anticoagulant Potential of Isorhamnetin and Its New Derivatives Isolated From Sea Buckthorn Fruits in Whole Blood

2021 ◽  
Author(s):  
Agata Rolnik ◽  
Bartosz Skalski ◽  
Jerzy Zuchowski ◽  
Anna Stochmal ◽  
Beata Olas
Keyword(s):  
Whole Blood ◽  
Thrombus Formation ◽  
Blood Platelets ◽  
Circulatory System ◽  
Sea Buckthorn ◽  
Phenolic Fraction ◽  
Adhesion Properties ◽  
Dietary Components ◽  
Analysis System

Abstract Blood platelets play a crucial role in hemostasis, the process responsible for keeping blood flowing in the circulatory system. However, unnecessary platelet activation can lead to aggregation at the site of atherosclerotic plaque rapture and the formation of a thrombus, which promotes atherothrombotic diseases. Various dietary components, such as phenolic compounds, are known to demonstrate antiplatelet and anticoagulant properties, and it is possible that these could form an important element in the prophylaxis and therapy of cardiovascular diseases. Our present study examines the biological activity of isorhamnetin (compound 1) and two isorhamnetin derivatives, compound 2 (3-O-beta-glucoside-7-O-alpha-rhamnoside) and compound 3 (3-O-beta-glucoside-7-O-alpha-(3”’-isovaleryl)-rhamnoside), isolated from the phenolic fraction of sea buckthorn fruit, against human washed blood platelets and human whole blood in vitro. The anti-platelet and anticoagulant potential was determined using (A) flow cytometry, (B) the thrombus-formation analysis system (T-TAS) and (C) colorimetry. The tested flavonoids demonstrated anticoagulant and anti-platelet potential, including anti-adhesive activity, with these effects being more intense in compound 2 than isorhamnetin. Compound 2 inhibited GPIIb/IIIa and P-selectin expression on blood platelets from whole blood, and demonstrated anti-adhesion properties in washed blood platelets and anti-coagulant potential in whole blood, measured by T-TAS.

scholarly journals Anti-Platelet Properties of Phenolic and Nonpolar FractionsIsolated from Various Organs of Elaeagnus rhamnoides (L.) A. Nelson in Whole Blood

2021 ◽  
Vol 22 (6) ◽  
pp. 3282
Author(s):  
Bartosz Skalski ◽  
Joanna Rywaniak ◽  
Aleksandra Szustka ◽  
Jerzy Żuchowski ◽  
Anna Stochmal ◽  
...  
Keyword(s):  
Whole Blood ◽  
Thrombus Formation ◽  
Blood Platelets ◽  
Sea Buckthorn ◽  
Platelet Activity ◽  
Bioactive Substances ◽  
Reducing Conditions ◽  
Triterpenic Acids ◽  
Platelet Proteome ◽  
Analysis System

Sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson) is a shrub growing in coastal areas. Its organs contain a range of bioactive substances including vitamins, fatty acids, various micro and macro elements, as well as phenolic compounds. Numerous studies of sea buckthorn have found it to have anticancer, anti-ulcer, hepatoprotective, antibacterial, and antiviral properties. Some studies suggest that it also affects the hemostasis system. The aim of the study was to determine the effect of six polyphenols rich and triterpenic acids rich fractions (A–F), taken from various organs of sea buckthorn, on the activation of blood platelets using whole blood, and to assess the effect of the tested fractions on platelet proteins: fraction A (polyphenols rich fraction from fruits), fraction B (triterpenic acids rich fraction from fruits), fraction C (polyphenols rich fraction from leaves), fraction D (triterpenic acids rich fraction from leaves), fraction E (polyphenols rich fraction from twigs), and fraction F (triterpenic acids rich fraction from twigs). Hemostasis parameters were determined using flow cytometry and T-TAS (Total Thrombus-formation Analysis System). Additionally, electrophoresis was performed under reducing and non-reducing conditions. Although all tested fractions inhibit platelet activation, the greatest anti-platelet activity was demonstrated by fraction A, which was rich in flavonol glycosides. In addition, none of the tested fractions (A–F) caused any changes in the platelet proteome, and their anti-platelet potential is not dependent on the P2Y12 receptor.

ABO Mismatched Platelet, Plasma and Cryoprecipitate Transfusions and Bleeding in Transfused Surgical Patients.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4166-4166
Author(s):  
Neil Blumberg ◽  
Kelly F. Gettings ◽  
Joanna M. Heal
Keyword(s):  
Platelet Function ◽  
Whole Blood ◽  
Blood Platelets ◽  
Blood Groups ◽  
Surgical Patients ◽  
Red Cell ◽  
Group A ◽  
Red Cell Transfusion ◽  
Platelet Transfusions

Abstract Three observations led us to investigate whether infusion of ABO non-identical platelets might impair, rather than improve hemostasis. (1) Exposure of platelets to immune complexes or platelet specific antibody can interfere with platelet function in vitro (Thromb Haemost76: 774, 1996). (2) In surgical patients receiving similar numbers of platelet transfusions, those receiving ABO non-identical platelets require 50% more red cell transfusions (Transfusion41: 790, 2001). (3) Patients with acute leukemia receiving prophylactic platelet transfusions typically are reported with serious bleeding at a rate of 15–20%, yet the bleeding rate in patients receiving only ABO identical platelets is below 5% (BMC Blood Disorders4: 6, 2004). In this study, the number of red cell transfusions and clinical outcomes during March 2002-Feb 2003 for all surgical patients of blood groups B and AB (B/AB) who received platelet transfusions were compared with patients of blood groups O and A (O/A). Recipients of blood groups B/AB would not be expected to experience excess bleeding, as measured by red cell transfusions, compared with patients of blood groups O/A. However, because of the lower prevalence of blood groups B/AB in the donor blood supply B/AB recipients may more frequently receive ABO mismatched platelet transfusions. O/A surgical patients (n=281) who received platelet transfusions required a mean of 13 ± 13 (SD) red cell transfusions as compared with B/AB patients (n=54), who required 19 ± 25 red cells (p =0.0086). O/A patients also had shorter length of stay, mean = 25 ± 34 days as compared with B/AB patients at 36 ± 59 days (p =0.064). Rates of mortality and nosocomial infections were not statistically significantly different. O/A patients received a mean of 14 ± 19 units of whole blood platelets during and after surgery, compared with 16 ± 16 units for B/AB patients (p =0.47). O/A patients received a mean of 3.3 ± 6.2 ABO non-identical platelets in contrast with B/AB patients who received 7.5 ± 11 (p = 0.0001). Both groups received similar numbers of ABO identical platelets: 11 ± 16 (O/A) versus 9 ± 12 (B/AB) (p =0.35). All but two patients received only ABO identical FFP and both groups received similar total amounts of FFP (mean of 9 units versus 11 units). While O/A patients received similar mean amounts of cryoprecipitate (6 units) to B/AB patients (8 units), the B/AB patients received significantly more ABO non-identical cryoprecipitate (mean = 4.2 vs. 2.3 units; p = 0.02). To study the effects of ABO incompatible plasma on platelet function, we measured PFA-100 (epinephrine cartridge) closure times in reconstituted whole blood exposing group A platelets to either group A or O plasma. In four of seven instances, closure times for A platelets exposed to O plasma were prolonged by more than 50 seconds, compared with A platelets exposed to allogeneic A plasma. These preliminary results support previous observations that exposure to ABO non-identical platelet transfusions is associated with increased red cell transfusions. One possible mechanism is impaired platelet function caused by antibody or immune complex binding. We speculate that transfusion of ABO mismatched platelets, FFP and/or cryoprecipitate may in some instances exacerbate bleeding, rather than correcting defects in hemostasis. Though further investigation is needed before suggesting changes in clinical practice these findings raise the possibility that use of ABO identical blood components might reduce red cell transfusion needs in bleeding surgical patients.

Platelet ADP receptors contribute to the initiation of intravascular coagulation

Blood ◽  
2004 ◽  
Vol 103 (2) ◽  
pp. 594-600 ◽  
Author(s):  
Catherine Leon ◽  
Meike Alex ◽  
Antje Klocke ◽  
Eberhard Morgenstern ◽  
Christine Moosbauer ◽  
...  
Keyword(s):  
Whole Blood ◽  
Ex Vivo ◽  
Thrombus Formation ◽  
Intravascular Coagulation ◽  
Adp Receptors ◽  
Adp Receptor ◽  
Deficient Mice ◽  
Storage Pools

Abstract While the adenosine 5′-diphosphate (ADP) pathway is known to enhance thrombus formation by recruiting platelets and leukocytes to the primary layer of collagen-adhering platelets, its role for the initiation of coagulation has not been revealed. Ex vivo inhibition of the P2Y12 ADP receptor by clopidogrel administration diminished the rapid exposure of tissue factor (TF), the major initiator of coagulation, in conjugates of platelets with leukocytes established by the contact of whole blood with fibrillar collagen. Under in vitro conditions, the P2Y12 and P2Y1 ADP receptors were both found to be implicated in the exposure of TF in collagen-activated whole blood. Immunoelectron-microscopy revealed that collagen elicited the release of TF from its storage pools within the platelets. Functional activation of the intravascular TF was reduced by inhibition of the ADP receptors, partially due to the disruption of the platelet-neutrophil adhesions. Injection of collagen into the venous system of mice increased the number of thrombin-antithrombin complexes, indicative for the formation of thrombin in vivo. In P2Y1-deficient mice, the ability of collagen to enhance the generation of thrombin was impaired. In conclusion, the platelet ADP pathway supports the initiation of intravascular coagulation, which is likely to contribute to the concomitant formation of fibrin at the site of the growing thrombus.

CD44 sensitivity of platelet activation, membrane scrambling and adhesion under high arterial shear rates

2016 ◽  
Vol 115 (01) ◽  
pp. 99-108 ◽  
Author(s):  
Kousi Alzoubi ◽  
Madhumita Chatterjee ◽  
Britta Walker ◽  
Patrick Münzer ◽  
Dong Luo ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Thrombus Formation ◽  
Blood Platelets ◽  
Protein Abundance ◽  
Integrin Activation ◽  
Platelet Surface ◽  
Shear Rates ◽  
Occlusive Vascular Disease ◽  
Phosphatidylserine Exposure

SummaryCD44 is required for signalling of macrophage migration inhibitory factor (MIF), an anti-apoptotic pro-inflammatory cytokine. MIF is expressed and released from blood platelets, key players in the orchestration of occlusive vascular disease. Nothing is known about a role of CD44 in the regulation of platelet function. The present study thus explored whether CD44 modifies degranulation (P-selectin exposure), integrin activation, caspase activity, phosphatidylserine exposure on the platelet surface, platelet volume, Orai1 protein abundance and cytosolic Ca2+-activity ([Ca2+]i). Platelets from mice lacking CD44 (cd44-/- ) were compared to platelets from corresponding wild-type mice (cd44+/+ ). In resting platelets, P-selectin abundance, αllbβ3 inte-grin activation, caspase-3 activity and phosphatidylserine exposure were negligible in both genotypes and Orai1 protein abundance, [Ca2+]i, and volume were similar in cd44-/- and cd44+/+ platelets. Platelet degranulation and αllbβ3 integrin activation were significantly increased by thrombin (0.02 U/ml), collagen related peptide (CRP, 2 µg/ml and Ca2+-store depletion with thapsigargin (1 µM), effects more pronounced in cd44-/- than in cd44+/+ platelets. Thrombin (0.02 U/ml) increased platelet [Ca2+]i, caspase-3 activity, phosphatidylserine exposure and Orai1 surface abundance, effects again significantly stronger in cd44-/- than in cd44+/+ platelets. Thrombin further decreased forward scatter in cd44-/- and cd44+/+ platelets, an effect which tended to be again more pronounced in cd44-/- than in cd44+/+ platelets. Platelet adhesion and in vitro thrombus formation under high arterial shear rates (1,700 s-1) were significantly augmented in cd44-/- mice. In conclusion, genetic deficiency of CD44 augments activation, apoptosis and prothrombotic potential of platelets.

scholarly journals The Changes of Blood Platelet Reactivity in the Presence of Crude Extracts Isolated from Leaves and Twigs of Elaeagnus Rhamnoides (L.) A. Nelson Measuring in Whole Blood

2021 ◽  
Author(s):  
Bartosz Skalski ◽  
Joanna Rywaniak ◽  
Jerzy Żuchowski ◽  
Anna Stochmal ◽  
Beata Olas
Keyword(s):  
Phenolic Compounds ◽  
Platelet Activation ◽  
Whole Blood ◽  
Leaf Extract ◽  
Platelet Reactivity ◽  
Blood Platelets ◽  
Blood Platelet ◽  
Surface Expression ◽  
Sea Buckthorn ◽  
Crude Extracts

Abstract Uncontrolled blood platelet activation is an important risk factor of cardiovascular disease (CVDs). Various studies on phenolic compounds indicate that they have a protective effect on the cardiovascular system through different mechanisms, including the reduction of blood platelet activation. One of the plants that is particularly rich in phenolic compounds is sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson). The aim of the present study in vitro was to determine the anti-platelet properties of crude extracts isolated from leaves and twigs of E. rhamnoides (L.) A. Nelson in whole blood using flow cytometric and total thrombus-formation analysis system (T-TAS). The aim of our study was also analyze of blood platelet proteoms in the presence of different sea buckthorn extracts. A significant new finding is a decrease surface expression of P-selectin on blood platelets stimulated by 10 µM ADP and 10 µg/mL collagen, and a decrease surface expression of GPIIb/IIIa active complex on non-activated platelets and platelets stimulated by 10 µM ADP and 10 µg/mL collagen in the presence of sea buckthorn leaf extract (especially at the concentration 50 µg/mL). The twig extract also displayed antiplatelet potential. However, this activity was higher in the leaf extract than in the twig extract in whole blood. In addition, our present findings clearly demonstrate that investigated plant extracts have anticoagulant properties (measured by T-TAS). Therefore, the two tested extracts may be promising candidates for the natural anti-platelet and anticoagulant supplements.

scholarly journals Cardiovascular risk factors are associated with augmented thrombogenicity in healthy individuals: analysis using the Total Thrombus-formation Analysis System

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Yuu Oda ◽  
Takashi Ito ◽  
Yoichiro Yamada ◽  
Tadashi Koga ◽  
Tomoka Nagasato ◽  
...  
Keyword(s):  
Risk Factors ◽  
Cardiovascular Risk ◽  
Healthy Volunteers ◽  
Cardiovascular Risk Factors ◽  
Whole Blood ◽  
Thrombus Formation ◽  
Reference Intervals ◽  
Blood Samples ◽  
Whole Blood Samples ◽  
Analysis System

Abstract Background Rupture of an atherosclerotic plaque and subsequent exposure of the subendothelial prothrombotic matrix to blood cause arterial thrombosis. Circulating platelets play an indispensable role in the growth of arterial thrombi partially owing to their unique ability to adhere to the subendothelial matrix and to aggregate to each other under flow conditions. Recently, the Total Thrombus-formation Analysis System (T-TAS) was developed for ex vivo analysis of the thrombogenic potential of whole blood samples under flow conditions. Despite the potential clinical utility of the T-TAS in assessing the risk for thrombosis and bleeding, reference intervals for T-TAS analysis in healthy individuals have not been determined. Methods In total, 122 whole blood samples were collected from healthy volunteers ranging in age from 25 to 45 years. T-TAS analysis and hematological, physiological, and lifestyle assessments were conducted in these subjects. Whole blood samples anticoagulated with hirudin were perfused into a collagen-coated microchip (PL chip). The time to 10 kPa and the area under the flow pressure curve up to 10 min (AUC10) were analyzed as representative variables for thrombogenic potential. Reference intervals, which were defined as 2.5–97.5 percentiles, were determined. Additionally, univariate and multivariate analyses were performed to identify factors associated with the AUC10 in the T-TAS. Results The time to 10 kPa and the AUC10 widely varied, even in healthy volunteers. The reference intervals were 1.50–4.02 min and 223.4–456.8, respectively, at a shear rate of 1500 s− 1. Univariate and multivariate analyses showed that platelet counts were most significantly associated with the AUC10 of the T-TAS. The presence of one or more cardiovascular risk factors of a high body mass index, a high pulse pressure, high fasting serum glucose levels, high low-density lipoprotein-cholesterol levels, a history of smoking, and no habitual exercise, had the second largest effect on the AUC10 of the T-TAS. Conclusions Healthy volunteers who had any cardiovascular risk factors showed augmented thrombogenicity, even in artificial uniform capillaries, compared with those without any risk factors in the T-TAS.

IN VIVO CHANGES IN SYSTEMIC PCO2 AND PO2 INFLUENCE THE THROMBOEMBOLIC REACTION FOLLOWING WALL PUNCTURE IN VENULES BUT NOT IN ARTERIOLES

1987 ◽  
Author(s):  
Mirjam G A oude Egbrink ◽  
Geert Jan Tangelder ◽  
Dick W Slaaf ◽  
Robert S Reneman
Keyword(s):  
Thrombus Formation ◽  
Fluid Dynamic ◽  
Blood Platelets ◽  
Control Group ◽  
Blood Gas ◽  
Systemic Blood ◽  
Normal Ranges ◽  
Experimental Group

Changes in pH and PCO2 influence the aggregation of blood platelets in response to various agents in vitro. In the present study intravital video-microscopy was used to investigate whether changes in systemic blood gas values influence the thromboembolic reaction in vivo as induced by vessel wall injury.The microtrauma was induced by puncturing the walls of microvessels in the rabbit mesentery (diameter range: 20-40 μm) with glass micropipets (tip diameters: 6-8 μm). The thromboembolic reactions were compared in two groups of anesthetized rabbits. The control group was ventilated to keep the blood gas values within normal ranges (means: pH=7.40, pCO2=32.9 mmHg, pO2=104.7 mmHg). The experimental group breathed spontaneously (mean blood gas values: pH=7.34, pCO2=50.5 mmHg, pO2=48.1 mmHg). The pCO2 and pO2 values were significantly different between both groups.In arterioles and venules of both groups bleeding and thrombus formation started immediately following wall puncture. Bleeding times were short (medians between 1.0 and 2.6 s). Parts of the thrombi started to embolize between 11.4 and 18.2 s following wall puncture (medians). In the control group embolization continued for 101 s in the arterioles and 17 s in the venules; during these periods 6 and 1 emboli were produced, respectively (all median values). In the experimental group the duration of embolization in the arterioles was 143 s in which period 7.5 emboli were produced, values not significantly different from control. In the venules of the experimental group embolization and hence platelet reaction went on uninhibited during the whole observation period of 600 s and 30 emboli were produced. Fluid dynamic factors cannot explain the differences in thromboembolic reaction between the control and experimental venules; vessel diameters and red blood cell velocities were not significantly different between both groups. Therefore, it is likely that the change in thromboembolic reaction in the venules results from the changes in systemic PCO2 and/or pO2. The different reactions in arterioles and venules in response to the altered systemic blood gas values might arise from different reactions in the vessel walls.

The influence of different rewetting procedures on the thrombogenicity of nanoporous poly(ether imide) microparticles

2020 ◽  
pp. 1-14
Author(s):  
S. Braune ◽  
J. Bäckemo ◽  
S. Lau ◽  
M. Heuchel ◽  
K. Kratz ◽  
...  
Keyword(s):  
Platelet Activation ◽  
Thrombus Formation ◽  
Blood Platelets ◽  
Saline Group ◽  
Vinyl Pyrrolidone ◽  
Covalent Attachment ◽  
Reference Ranges ◽  
Chemical Treatments ◽  
Phosphate Buffered Saline

Nanoporous microparticles prepared from poly(ether imide) (PEI) are discussed as candidate adsorber materials for the removal of uremic toxins during apheresis. Polymers exhibiting such porosity can induce the formation of micro-gas/air pockets when exposed to fluids. Such air presenting material surfaces are reported to induce platelet activation and thrombus formation. Physical or chemical treatments prior to implantation are discussed to reduce the formation of such gas nuclei. Here, we report about the influence of different rewetting procedures – as chemical treatments with solvents – on the thrombogenicity of hydrophobic PEI microparticles and PEI microparticles hydrophilized by covalent attachment of poly(vinyl pyrrolidone) (PVP) of two different chain lengths. Autoclaved dry PEI particles of all types with a diameter range of 200 – 250 μm and a porosity of about 84% ±2% were either rewetted directly with phosphate buffered saline (24 h) or after immersion in an ethanol-series. Thrombogenicity of the particles was studied in vitro using human sodium citrated whole blood (60 min, 5 rpm vertical rotation). Numbers of non-adherent platelets were quantified, and adhesion of blood cells was qualitatively analyzed by bright field microscopy. Platelet activation (percentage of CD62P positive platelets and amounts of soluble P-Selectin) and platelet function (PFA100 closure times) were analysed. Retention of blood platelets on the particles was similar for all particle types and both rewetting procedures. Non-adherent platelets were less activated after contact with ethanol-treated particles of all types compared to those rewetted with phosphate buffered saline as assessed by a reduced number of CD62P-positive platelets and reduced amounts of secreted P-Selectin (P < 0.05 each). Interestingly, the hydrophilic surfaces significantly increased the number of activated platelets compared to hydrophobic PEI regardless of the rewetting agent. This suggests that, apart from wettability, other material properties might be more important to regulate platelet activation. PFA100 closure times were reduced and within the reference ranges in the ethanol group, however, significantly increased in the saline group. No substantial difference was detected between the tested surface modifications. In summary, rewetting with ethanol resulted in a reduced thrombogenicity of all studied microparticles regardless of their wettability, most likely resulting from the evacuation of air from the nanoporous particles.

scholarly journals Evaluation of the Antithrombotic Effects of Rivaroxaban and Apixaban Using the Total Thrombus-Formation Analysis System®: In Vitro and Ex Vivo Studies

2016 ◽  
Vol 8 (12) ◽  
pp. 899-907 ◽  
Author(s):  
Hidekazu Sugihara ◽  
Yoshiaki Idemoto ◽  
Takashi Kuwano ◽  
Yoshihisa Nagata ◽  
Joji Morii ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Thrombus Formation ◽  
Antithrombotic Effects ◽  
Analysis System

scholarly journals Preparations from Selected Cucurbit Vegetables as Antiplatelet Agents – An in Vitro Study

2021 ◽  
Author(s):  
Agata Rolnik ◽  
Bartosz Skalski ◽  
Anna Stochmal ◽  
Beata Olas
Keyword(s):  
Platelet Activation ◽  
Cucurbita Pepo ◽  
Thrombus Formation ◽  
Blood Platelets ◽  
In Vitro Study ◽  
Blood Platelet ◽  
Arachidonic Acid Metabolism ◽  
Platelet Activity ◽  
Activated Platelets

Abstract Increased blood platelet activation plays an important role in cardiovascular diseases (CVDs). Recent experiments indicate that certain fruits and vegetables, including onion, garlic, and beetroot, have anti-platelet potential and therefore may reduce the likelihood of CVDs. While vegetables from the Cucuritaceae family are known to exerting beneficial antioxidant and anti-inflammatory effects, their effects on blood platelet activation are poorly understood. Therefore, the aim of the present study was to determine the effect on platelet adhesion of preparations from selected cucurbits: pumpkin (Cucirbita pepo; fruit without seeds), zucchini (Cucurbita pepo convar. giromontina; fruit with seeds), cucumber (Cucumis sativus; fruit with seeds), white pattypan squash (Cucurbita pepo var. patisoniana; fruit without seeds) and yellow pattypan squash (Cucurbita pepo var. patisoniana, fruit without seeds). It also evaluates the activity of these preparations on enzymatic lipid peroxidation in thrombin-activated washed blood platelets by TBARS assay. The study also determines the anti-platelet and anticoagulant properties of these five cucurbit preparations in whole blood by flow cytometry and with the total thrombus-formation analysis system (T-TAS) and evaluates the cytotoxicity of the tested preparations against platelets based on LDH activity. The results indicate that the yellow Cucurbita pepo var. patisoniana preparation demonstrated stronger anti-platelet properties than the other tested preparations, reducing the adhesion of thrombin-activated platelets to collagen/fibrinogen, and inhibiting arachidonic acid metabolism and GPIIb/IIIa expression on 10 µM ADP-activated platelets. None of the preparations was found to cause platelet lysis. Our findings provide new information on the anti-platelet activity of the tested cucurbit preparations and their potential for treating CVDs associated with platelet hyperactivity.

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