Effect of Progesterone and Synthetic Progestins on Whole Blood Clot Formation and Erythrocyte Structure

AbstractCombined oral contraceptive (COC) use is a risk factor for venous thrombosis (VT) and related to the specific type of progestin used. VT is accompanied by inflammation and pathophysiological clot formation, that includes aberrant erythrocytes and fibrin(ogen) interactions. In this paper, we aim to determine the influence of progesterone and different synthetic progestins found in COCs on the viscoelasticity of whole blood clots, as well as erythrocyte morphology and membrane ultrastructure, in an in vitro laboratory study. Thromboelastography (TEG), light microscopy, and scanning electron microscopy were our chosen methods. Our results point out that progestins influence the rate of whole blood clot formation. Alterations to erythrocyte morphology and membrane ultrastructure suggest the presence of eryptosis. We also note increased rouleaux formation, erythrocyte aggregation, and spontaneous fibrin formation in whole blood which may explain the increased risk of VT associated with COC use. Although not all COC users will experience a thrombotic event, individuals with a thrombotic predisposition, due to inflammatory or hematological illness, should be closely monitored to prevent pathological thrombosis.

Download Full-text

The Haemocompatibility of Biomaterials In Vitro: Investigations on the Mechanism of the Whole-blood Clot Formation Test

Alternatives to Laboratory Animals ◽

10.1177/026119299202000305 ◽

1992 ◽

Vol 20 (3) ◽

pp. 390-395 ◽

Cited By ~ 1

Author(s):

Thomas Groth ◽

Katrin Derdau ◽

Frank Strietzel ◽

Frank Foerster ◽

Hartmut Wolf

Keyword(s):

Whole Blood ◽

Blood Clotting ◽

Blood Clot ◽

Formation Rate ◽

Clot Formation ◽

Coagulation Cascade ◽

Contact Activation ◽

Human Fibrinogen ◽

Static Conditions

Twenty years ago Imai & Nose introduced a whole-blood clotting test for the estimation of haemocompatibility of biomaterials in vitro In our paper a modification of this assay is described and the mechanism of clot formation further elucidated. It was found that neither the inhibition of platelet function nor the removal of platelets from blood significantly changed the clot formation rate on glass and polyvinyl chloride in comparison to the rate tor whole blood. Scanning electron microscopy demonstrated that platelets were not involved in clot formation near the blood/biomaterial interface. Thus, it was concluded that the system of contact activation of the coagulation cascade dominates during clot formation under static conditions. The latter conclusion was supported by the fact that preadsorption of human serum albumin or human fibrinogen onto the glass plates used, decreased the clot formation rate in the same manner.

Download Full-text

Potential inheritance patterns of a prothrombin gene mutation in a 23-year-old female and ethical considerations of a positive diagnosis: a case report

Dentistry 3000 ◽

10.5195/d3000.2017.69 ◽

2017 ◽

Vol 5 (1) ◽

Author(s):

Hannah Brand

Keyword(s):

Gene Mutation ◽

Blood Clot ◽

Clot Formation ◽

Ethical Implications ◽

Increased Risk ◽

Blood Clots ◽

History Of ◽

Prothrombin Gene Mutation ◽

Prothrombin Gene ◽

The Individual

Background: Prothrombin, also called Factor II, is a blood clotting protein found in all individuals that is necessary to form blood clots. In most individuals, a balance between bleeding and blood clot formation occurs. However, in individuals with a mutation in the prothrombin gene, the balance is disrupted due to excess production of prothrombin which leads to an increase in blood clot formation (1). Inherited predispositions to blood clot formation are termed hereditary thrombophilia. Prothrombin G20210A mutations are one of the most common hereditary gene associations. Case Description: This report examines the case of a 23-year-old female who has tested positive for the prothrombin gene mutation. The individual has an extensive history of blood clots including 8 deep vein thromboses (DVTs), 4 pulmonary embolisms with one complete infarction, 4 superficial clots, and a miscarried pregnancy attributed to her thrombophilia. The individual has a significant family history of the mutation and takes Coumadin daily for prevention of further clots. Practical and Ethical Implications: Despite a strong familial history of blood clots and related hospitalizations, the parents of the case individual do not want to get their other children tested for the mutation for fear that the children will be denied insurance coverage in their futures. The case individual will likely continue to be on blood thinners indefinitely. This will affect many aspects of her life, including dental treatment, as she will be at an increased risk for bleeding.

Download Full-text

Influence of Factor Xllla Activity on Human Whole Blood Clot Lysis In Vitro

Thrombosis and Haemostasis ◽

10.1055/s-0038-1651088 ◽

1987 ◽

Vol 57 (02) ◽

pp. 171-175 ◽

Cited By ~ 44

Author(s):

J W C M Jansen ◽

F Haverkate ◽

J Koopman ◽

H K Nieuwenhuis ◽

C Kluft ◽

...

Keyword(s):

Whole Blood ◽

Major Part ◽

Blood Clot ◽

Tissue Type ◽

Clot Lysis ◽

Type Plasminogen Activator ◽

Blood Clots ◽

Lysis Rate ◽

Per Se

SummaryWe studied the influence of Factor XIII a (F XIII a) activity on the lysis rate of fresh whole human blood clots, without using anticoagulants. Clotting was induced by exogenous thrombin, lysis by tissue-type Plasminogen Activator (t-PA) added before clotting. After various periods of time, lysis rates were determined by measuring the radioactivity in the supernatant of the clot originating from 125I-Fibrinogen added before clotting.Lysis rates were determined in the presence of endogenous F XHIa and compared with those obtained after specific inhibition of F XIII a activity. We used an IgG fraction of an antiserum quenching the F XIII a activity. Addition of increasing amounts of the antibodies to normal blood resulted in a dramatic increase in clot lysis rate, concomitant with loss of F XIII activity. Lysis of blood clots from a patient with a congenital, homozygous, functional α2-Antiplasmin (α2-AP) deficiency (α2-AP-Enschede) was not or slightly increased by the anti F XIII antibodies indicating that fibrin-fibrin crosslinking per se does not contribute essentially to resistance of the blood clot against fibrinolysis. Both active α2-AP and F XIII a are required for the major part of the F XIII-dependent resistance of whole blood clots against lysis.

Download Full-text

Studies on Thrombolysis with Streptokinase

Thrombosis and Haemostasis ◽

10.1055/s-0038-1654310 ◽

1971 ◽

Vol 25 (02) ◽

pp. 354-378 ◽

Cited By ~ 5

Author(s):

R Gottlob ◽

L Stockinger ◽

U Pötting ◽

G Schattenmann

Keyword(s):

Electron Microscopy ◽

Cross Section ◽

Whole Blood ◽

Jugular Vein ◽

Thin Sections ◽

The Third ◽

Morphological Findings ◽

Blood Clots ◽

Arteries And Veins

SummaryIn vitro whole blood clots of various ages, experimental thrombi produced in the jugular vein of rabbits and human thrombi from arteries and veins were examined in semi-thin sections and by means of electron microscopy.In all types of clots examined a typical course of retraction was found. Retraction starts with a dense excentrical focus which grows into a densification ring. After 24 hours the entire clot becomes almost homogeneously dense; later a secondary swelling sets in.Shortly after coagulation the erythrocytes on the rim of the clot are bi-concave discs. They then assume the shape of crenate spheres, turn into smooth spheres and finally become indented ghosts which have lost the largest part of their contents. In the inner zone, which makes up the bulk of the clot, we observed bi-concave discs prior to retraction. After retraction we see no crenations but irregularly shaped erythrocytes. Once the secondary swelling sets in, the cross-section becomes polygonal and later spherical. After extensive hemolysis we observe the “retiform thrombus” made up of ghosts.Experimental and clinical thrombi present the same morphology but are differentiated from in vitro clots by: earlier hemolysis, immigration of leukocytes, formation of a rim layer consisting of fibrin and thrombocytes, and the symptoms of organization. Such symptoms of organization which definitely will prevent lysis with streptokinase were found relatively late in experimental and clinical thrombi. Capillary buds and capillary loops were never found in clinical thrombi prior to the third month.The morphological findings agree with earlier physical and enzymatic investigations. The observation that phenomena of reorganization occur relatively late and frequently only in the rim areas of large thrombi explains why lytic therapy is possible in some of the chronic obliterations.

Download Full-text

TITANIUM NANOPARTICLES CONJUGATED WITH STREPTOKINASE AS A MODIFIED THROMBOLYTIC AGENT

International Journal of Current Pharmaceutical Research ◽

10.22159/ijcpr.2020v12i1.36824 ◽

2020 ◽

pp. 18-25

Author(s):

HAYDER H. ABED ◽

ESTABRAQ AR. ALWASITI ◽

AMIR T. TAWFEEQ

Keyword(s):

Thrombolytic Agent ◽

Blood Clot ◽

Control Group ◽

Thrombolytic Activity ◽

Protein Assay ◽

Blood Clots ◽

Ft Ir ◽

Titanium Nanoparticles ◽

D Dimer

Objective: Blood clots are the main cause of death worldwide by stroke and myocardial infarction. Streptokinase a thrombolytic agent that is used in the treatment of circulatory disorders. Methods: Titanium Nanoparticles was supplied from Changsha Santech Co. Its characterized were studied using (FT-IR, XRD, AFM, FE-SEM). Streptokinase at concentration 0.1 mg/ml was conjugated with Titanium nanoparticles using PH equal to 5.2 with continuous stirring. Formation of Streptokinase loading Titanium nanoparticles confirmed using FT-IR, Ninhydrine’s test and Bradford protein assay. Physicochemical Properties were studied in vitro. Thrombolytic activity in vitro was determined using d–dimer indicator and weight of blood clot after treatment as indicators of thrombolytic activity. Results: Titanium nanoparticles show particle size at range 31 nm. The thrombolytic activity of streptokinase loading Titanium nanoparticles shows significant value in d-dimer and weight of blood clot compared with the control group and non-significant compared with an equivalent amount of streptokinase alone. Conclusion: Titanium nanoparticles conjugated with streptokinase show high thrombolytic activity against blood clots in vitro.

Download Full-text

Studies of T-lymphocytes in preleukemic disorders and acute nonlymphocytic leukemia: in vitro radiosensitivity, mitogenic responsiveness, colony formation, and enumeration of lymphocytic subpopulations

Blood ◽

10.1182/blood.v61.3.449.449 ◽

1983 ◽

Vol 61 (3) ◽

pp. 449-455 ◽

Cited By ~ 30

Author(s):

SJ Knox ◽

BR Greenberg ◽

RW Anderson ◽

LS Rosenblatt

Keyword(s):

Whole Blood ◽

Colony Formation ◽

Tritiated Thymidine ◽

Lymphocyte Stimulation Test ◽

Acute Nonlymphocytic Leukemia ◽

Clinically Normal ◽

Normal Individuals ◽

Increased Risk ◽

X Irradiation

Abstract Tritiated thymidine incorporation in a whole blood lymphocyte stimulation test (LST) and lymphocyte colony formation (CFU-L) from whole blood were measured following in vitro x-irradiation. Lymphocytes from patients with myelodysplastic disorders, acute nonlymphocytic leukemia, and patients at increased risk for leukemia because of their primary disease and/or cytotoxic therapy were found to be significantly more sensitive to in vitro x-irradiation than lymphocytes from clinically normal individuals. Cloning efficiencies and mitogenic responsiveness of patient lymphocytes were significantly depressed as compared to normal values. Using monoclonal antibodies to specific surface markers, quantitative abnormalities in lymphocytic subpopulations from myelodysplastic patients also were observed. These findings are suggestive of a defect at the T-cell level that may directly or indirectly affect hematopoiesis.

Download Full-text

Thrombin generation and fibrin clot formation under hypothermic conditions: An in vitro evaluation of tissue factor initiated whole blood coagulation

Journal of Critical Care ◽

10.1016/j.jcrc.2013.10.010 ◽

2014 ◽

Vol 29 (1) ◽

pp. 24-30 ◽

Cited By ~ 9

Author(s):

Matthew F. Whelihan ◽

Armin Kiankhooy ◽

Kathleen E. Brummel-Ziedins

Keyword(s):

Tissue Factor ◽

Blood Coagulation ◽

Whole Blood ◽

Thrombin Generation ◽

Fibrin Clot ◽

Clot Formation ◽

In Vitro Evaluation

Download Full-text

In vitro whole blood clot lysis by rt-PA is inversely correlated with baseline plasma levels of t-PA antigen

Fibrinolysis and Proteolysis ◽

10.1016/0268-9499(94)90401-4 ◽

1994 ◽

Vol 8 ◽

pp. 43

Author(s):

M. Colucci ◽

S. Scopece ◽

A. Gelato ◽

L.G. Cavallo ◽

N. Semeraro

Keyword(s):

Whole Blood ◽

Plasma Levels ◽

Blood Clot ◽

Clot Lysis

Download Full-text

Role of factor XIII in fibrin clot formation and effects of genetic polymorphisms

Blood ◽

10.1182/blood.v100.3.743 ◽

2002 ◽

Vol 100 (3) ◽

pp. 743-754 ◽

Cited By ~ 243

Author(s):

Robert A. S. Ariëns ◽

Thung-Shenq Lai ◽

John W. Weisel ◽

Charles S. Greenberg ◽

Peter J. Grant

Keyword(s):

Genetic Polymorphisms ◽

Factor Xiii ◽

Blood Clot ◽

Site Directed Mutagenesis ◽

Clot Formation ◽

Factor Xiiia ◽

Clotting Factors ◽

Functional Features

Abstract Factor XIII and fibrinogen are unusual among clotting factors in that neither is a serine protease. Fibrin is the main protein constituent of the blood clot, which is stabilized by factor XIIIa through an amide or isopeptide bond that ligates adjacent fibrin monomers. Many of the structural and functional features of factor XIII and fibrin(ogen) have been elucidated by protein and gene analysis, site-directed mutagenesis, and x-ray crystallography. However, some of the molecular aspects involved in the complex processes of insoluble fibrin formation in vivo and in vitro remain unresolved. The findings of a relationship between fibrinogen, factor XIII, and cardiovascular or other thrombotic disorders have focused much attention on these 2 proteins. Of particular interest are associations between common variations in the genes of factor XIII and altered risk profiles for thrombosis. Although there is much debate regarding these observations, the implications for our understanding of clot formation and therapeutic intervention may be of major importance. In this review, we have summarized recent findings on the structure and function of factor XIII. This is followed by a review of the effects of genetic polymorphisms on protein structure/function and their relationship to disease.

Download Full-text