Evaluation of the hypercoagulability markers and global haemostatic tests and global haemostatic tests in pregnancies complicated with pre-eclampsia

Preeklampsija je multisistemski poremećaj koji se javlja kao komplikacija trudnoće i predstavlja jedan od vodećih uzroka mortaliteta i morbiditeta majki, ali i njihovog potomstva. Smatra se da preeklampsija nastaje kao posledica poremećaja u ranom razvoju placente koji dovodi do aktivacije maternalnog vaskularnog endotela što rezultuje generalizovanom vazokonstrikcijom, značajnim metaboličkim promenama, disfunkcijom endotela, pojačanim inflamatornim odgovorom i aktivacijom koagulacije. TakoĎe, dokazano je da pojava preeklampsije u trudnoći ukazuje na postojanje nepovoljnog kardio-metaboličkog profila ţene, te je u novijim vodičima preeklampsija uvrštena u dugoročne faktore rizika za razvoj kardiovaskularnih bolesti (KVB). Pored toga, preeklampsija i KVB imaju zajedničke faktore rizika, a oba poremećaja su okarakterisana i zajedničkim patofiziološkim promenama. Klinički simptomi preeklampsije, hipertenzija i proteinurija, ispoljavaju se nakon 20. nedelje gestacije, mada dijagnoza preeklampsije moţe da se postavi i u odsustvu proteinurije ukoliko je novonastala hipertenzija udruţena sa pojavom trombocitopenije, poremećene funkcije jetre, novorazvijene bubreţne insuficijencije, plućnog edema ili novonastalih vizuelnih odnosno cerebralnih poremećaja, te uteroplacentalnog poremećaja koji rezultuje zastojem u rastu ploda. S obzirom da je hiperkoagulabilno stanje u preeklampsiji dodatno pojačano u odnosu na normalnu trudnoću cilj ove studije je bio da se ispita korisnost globalnih testova hemostaze, endogenog trombinskog potencijala i ukupnog hemostatskog potencijala, u proceni poremećaja hemostaze u preeklampsiji, pre i nakon poroĎaja, kao i da se utvrdi povezanost ispitivanih parametara sa ishodima preeklamptične trudnoće. Analizirane su takoĎe i karakteristike fibrinskih ugrušaka u normalnoj i preeklamptičnoj trudnoći. Pored toga, odreĎivani su različiti fenotipovi ekstracelularnih vezikula i analizirana je njihova povezanost sa ispitivanim hemostatskim parametrima. U studiju je uključeno 46 trudnica sa preeklampsijom i 80 zdravih trudnica...Pre-eclampsia is a multisystem disorder that occurs as a complication of pregnancy and is one of the leading causes of maternal mortality and morbidity, as well as of their offspring. The major cause of pre-eclampsia is considered to be a defect in early placental development leading to maternal vascular endothelial activation which results in generalized vasoconstriction, significant metabolic changes, endothelial dysfunction, enhanced inflammatory response, and coagulation activation. Also, it has been proven that the occurrence of pre-eclampsia in pregnancy indicates the existence of an unfavourable cardio- metabolic profile in women, and recent guidelines include pre-eclampsia as a long-term risk factor for the development of cardiovascular disease (CVD). Moreover, pre-eclampsia and CVD share common risk factors, and both disorders are characterized by common pathophysiological changes. Clinical symptoms of preeclampsia, hypertension and proteinuria, can be seen after 20 weeks of gestation, although the diagnosis of pre-eclampsia can be made in the absence of proteinuria if the new-onset hypertension is associated with thrombocytopenia, impaired liver function, new-onset renal failure, pulmonary oedema or new-onset visual or cerebral disorders, and uteroplacental disorder that results in fetal growth retardation. Bearing in mind that the hypercoagulable state in pre-eclampsia is further enhanced compared to normal pregnancy, the aim of this study was to evaluate the usefulness of global haemostatic assays, endogenous thrombin potential and overall haemostatic potential in the assessment of the haemostatic disorders in pre-eclampsia before and after delivery and to analyse the results of these assays in relation to the outcomes of preeclamptic pregnancy. The characteristics of fibrin clots in normal and preeclamptic pregnancies were also analyzed. Furthermore, different phenotypes of extracellular vesicles were determined and their association with the investigated hemostatic parameters was analyzed. The study involved 46 pregnant women with pre-eclampsia and 80 healthy pregnant women..

Evaluation of the hypercoagulability markers and global haemostatic tests and global haemostatic tests in pregnancies complicated with pre-eclampsia

Preeklampsija je multisistemski poremećaj koji se javlja kao komplikacija trudnoće ipredstavlja jedan od vodećih uzroka mortaliteta i morbiditeta majki, ali i njihovog potomstva.Smatra se da preeklampsija nastaje kao posledica poremećaja u ranom razvoju placente kojidovodi do aktivacije maternalnog vaskularnog endotela što rezultuje generalizovanomvazokonstrikcijom, značajnim metaboličkim promenama, disfunkcijom endotela, pojačaniminflamatornim odgovorom i aktivacijom koagulacije. TakoĎe, dokazano je da pojavapreeklampsije u trudnoći ukazuje na postojanje nepovoljnog kardio-metaboličkog profilaţene, te je u novijim vodičima preeklampsija uvrštena u dugoročne faktore rizika za razvojkardiovaskularnih bolesti (KVB). Pored toga, preeklampsija i KVB imaju zajedničke faktorerizika, a oba poremećaja su okarakterisana i zajedničkim patofiziološkim promenama.Klinički simptomi preeklampsije, hipertenzija i proteinurija, ispoljavaju se nakon 20. nedeljegestacije, mada dijagnoza preeklampsije moţe da se postavi i u odsustvu proteinurije ukolikoje novonastala hipertenzija udruţena sa pojavom trombocitopenije, poremećene funkcijejetre, novorazvijene bubreţne insuficijencije, plućnog edema ili novonastalih vizuelnihodnosno cerebralnih poremećaja, te uteroplacentalnog poremećaja koji rezultuje zastojem urastu ploda.S obzirom da je hiperkoagulabilno stanje u preeklampsiji dodatno pojačano u odnosuna normalnu trudnoću cilj ove studije je bio da se ispita korisnost globalnih testovahemostaze, endogenog trombinskog potencijala i ukupnog hemostatskog potencijala, uproceni poremećaja hemostaze u preeklampsiji, pre i nakon poroĎaja, kao i da se utvrdipovezanost ispitivanih parametara sa ishodima preeklamptične trudnoće. Analizirane sutakoĎe i karakteristike fibrinskih ugrušaka u normalnoj i preeklamptičnoj trudnoći. Poredtoga, odreĎivani su različiti fenotipovi ekstracelularnih vezikula i analizirana je njihovapovezanost sa ispitivanim hemostatskim parametrima.U studiju je uključeno 46 trudnica sa preeklampsijom i 80 zdravih trudnica...Pre-eclampsia is a multisystem disorder that occurs as a complication of pregnancyand is one of the leading causes of maternal mortality and morbidity, as well as of theiroffspring. The major cause of pre-eclampsia is considered to be a defect in early placentaldevelopment leading to maternal vascular endothelial activation which results in generalizedvasoconstriction, significant metabolic changes, endothelial dysfunction, enhancedinflammatory response, and coagulation activation. Also, it has been proven that theoccurrence of pre-eclampsia in pregnancy indicates the existence of an unfavourable cardio-metabolic profile in women, and recent guidelines include pre-eclampsia as a long-term riskfactor for the development of cardiovascular disease (CVD). Moreover, pre-eclampsia andCVD share common risk factors, and both disorders are characterized by commonpathophysiological changes. Clinical symptoms of preeclampsia, hypertension andproteinuria, can be seen after 20 weeks of gestation, although the diagnosis of pre-eclampsiacan be made in the absence of proteinuria if the new-onset hypertension is associated withthrombocytopenia, impaired liver function, new-onset renal failure, pulmonary oedema ornew-onset visual or cerebral disorders, and uteroplacental disorder that results in fetal growthretardation.Bearing in mind that the hypercoagulable state in pre-eclampsia is further enhancedcompared to normal pregnancy, the aim of this study was to evaluate the usefulness of globalhaemostatic assays, endogenous thrombin potential and overall haemostatic potential in theassessment of the haemostatic disorders in pre-eclampsia before and after delivery and toanalyse the results of these assays in relation to the outcomes of preeclamptic pregnancy. Thecharacteristics of fibrin clots in normal and preeclamptic pregnancies were also analyzed.Furthermore, different phenotypes of extracellular vesicles were determined and theirassociation with the investigated hemostatic parameters was analyzed.The study involved 46 pregnant women with pre-eclampsia and 80 healthy pregnantwomen..

Uticaj DOAK i DOAC-REMOVE® na testove koagulacije u toku testiranja trombofilije kod bolesnika lečenih primenom DOAK

Background/Aim. Direct oral anticoagulants (DOACs) administration significantly interferes with coagulation as-says. The aim of the study was to evaluate the effect of DOACs and DOAC-Remove® on coagulation assays dur-ing thrombophilia testing. Methods. The study was car-ried out from January 2019 to the end of June 2020. It in-cluded 30 DOAC-treated patients, 14 females and 16 males aged 23 to 63 (median age 47.6 years), tested for thrombophilia due to venous thromboembolism (VTE). Thrombophilia testing was performed using DOAC-Remove® tablets (activated charcoal). The results before and after DOAC-Remove® were compared. Results. Posi-tive lupus anticoagulant (LA) results were observed in 20% apixaban, 100% dabigatran, and 70% rivaroxaban-treated patients, while in samples after DOAC-Remove®, the LA positivity was observed only in one from the apix-aban group. Before DOAC-Remove®, the activated pro-tein C (APC) resistance (APC-R) was measurable in 40% dabigatran and 80% rivaroxaban-treated patients, while, after using DOAC-Remove®, the APC-R was measurable in all cases. Comparing the results obtained from the sam-ples before and after DOAC-Remove®, a difference was noted in relation to all dilute Russell's viper venom time (dRVVT) coagulation tests, except for the dRVVT ratio in the apixaban group. Clot-based methods for detecting the APC resistance were significantly affected by dabigatran and less by rivaroxaban. Conclusion. DOACs were prac-tically inactivated after the addition of the DOAC-Remove®, which made it possible to perform analyses for the LA and APC-R testing freely and obtain relevant re-sults.Uvod/Cilj. Primena direktnih oralnih antikoagulansa (DOAK) značajno utiče na testove koagulacije. Cilj rada bio je da se pro- ceni uticaj DOAK i DOAC-Remove® tableta (aktivni ugalj) na testove koagulacije tokom ispitivanja trombofilije. Metode. Istraživanjem, sprovedenim od januara 2019. do juna 2020. godine, obuhvaćeno je 30 bolesnika lečenih DOAK-om i testiranih na trombofiliju zbog venskog tromboembolzma (VTE). Bilo je 14 žena i 16 muškaraca, starosti od 23 do 63 godine (medijana 47,6 godina). Ispitivanje trombofilije izvršeno je upotrebom DOAC-Remove® tableta (aktivni ugalj). Upoređivani su rezultati pre i posle primene DOAC-Remove®. Rezultati. Pozitivni rezultati za lupus antikoagulantni (LA) test dobijeni su kod 20% bolesnika lečenih apiksabanom, kod 100% bolesnika lečenih dabigatranom i kod 70% lečenih riva- roksabanom, a u uzorcima posle DOAC-Remove® pozitivnost na LA dobijena je samo kod jednog bolesnika iz grupe lečnih apiksabanom. Pre primene DOAC-Remove®, rezistencija na aktivisani protein C (activated protein C resistance – APC-R) bila je merljiva kod 40% i 80% bolesnika lečenih dabigatranom, od- nosno rivaroksabanom, dok je posle primene DOAC- Remove®, APC-R bila merljiva u svim slučajevima. Upoređivanjem rezultata dobijenih iz uzoraka pre i posle primene DOAC-Remove®, primećena je razlika u odnosu na sve testove vremena koagulacije izvršene razblaženim Russell- ovim zmijskim otrovom (dilute Russell’s viper venom time – dRVVT), osim dRVVT u grupi bolesnika lečenih apiksabanom. Na koagulacionu metodu za otkrivanje APC-R značajno je uti- cao dabigatran, a manje rivaroksaban. Zaključak. Nakon primene DOAC-Remove® tableta, DOAK su praktično inaktivisani što je omogućilo izvođenje analiza za LA i APC-R i dobijanje relevantnih rezultata testova

Evaluation of the hypercoagulability markers and global haemostatic tests and global haemostatic tests in pregnancies complicated with pre-eclampsia

Preeklampsija je multisistemski poremećaj koji se javlja kao komplikacija trudnoće i predstavlja jedan od vodećih uzroka mortaliteta i morbiditeta majki, ali i njihovog potomstva. Smatra se da preeklampsija nastaje kao posledica poremećaja u ranom razvoju placente koji dovodi do aktivacije maternalnog vaskularnog endotela što rezultuje generalizovanom vazokonstrikcijom, značajnim metaboličkim promenama, disfunkcijom endotela, pojačanim inflamatornim odgovorom i aktivacijom koagulacije. TakoĎe, dokazano je da pojava preeklampsije u trudnoći ukazuje na postojanje nepovoljnog kardio-metaboličkog profila ţene, te je u novijim vodičima preeklampsija uvrštena u dugoročne faktore rizika za razvoj kardiovaskularnih bolesti (KVB). Pored toga, preeklampsija i KVB imaju zajedničke faktore rizika, a oba poremećaja su okarakterisana i zajedničkim patofiziološkim promenama. Klinički simptomi preeklampsije, hipertenzija i proteinurija, ispoljavaju se nakon 20. nedelje gestacije, mada dijagnoza preeklampsije moţe da se postavi i u odsustvu proteinurije ukoliko je novonastala hipertenzija udruţena sa pojavom trombocitopenije, poremećene funkcije jetre, novorazvijene bubreţne insuficijencije, plućnog edema ili novonastalih vizuelnih odnosno cerebralnih poremećaja, te uteroplacentalnog poremećaja koji rezultuje zastojem u rastu ploda. S obzirom da je hiperkoagulabilno stanje u preeklampsiji dodatno pojačano u odnosu na normalnu trudnoću cilj ove studije je bio da se ispita korisnost globalnih testova hemostaze, endogenog trombinskog potencijala i ukupnog hemostatskog potencijala, u proceni poremećaja hemostaze u preeklampsiji, pre i nakon poroĎaja, kao i da se utvrdi povezanost ispitivanih parametara sa ishodima preeklamptične trudnoće. Analizirane su takoĎe i karakteristike fibrinskih ugrušaka u normalnoj i preeklamptičnoj trudnoći. Pored toga, odreĎivani su različiti fenotipovi ekstracelularnih vezikula i analizirana je njihova povezanost sa ispitivanim hemostatskim parametrima. U studiju je uključeno 46 trudnica sa preeklampsijom i 80 zdravih trudnica...Pre-eclampsia is a multisystem disorder that occurs as a complication of pregnancy and is one of the leading causes of maternal mortality and morbidity, as well as of their offspring. The major cause of pre-eclampsia is considered to be a defect in early placental development leading to maternal vascular endothelial activation which results in generalized vasoconstriction, significant metabolic changes, endothelial dysfunction, enhanced inflammatory response, and coagulation activation. Also, it has been proven that the occurrence of pre-eclampsia in pregnancy indicates the existence of an unfavourable cardio- metabolic profile in women, and recent guidelines include pre-eclampsia as a long-term risk factor for the development of cardiovascular disease (CVD). Moreover, pre-eclampsia and CVD share common risk factors, and both disorders are characterized by common pathophysiological changes. Clinical symptoms of preeclampsia, hypertension and proteinuria, can be seen after 20 weeks of gestation, although the diagnosis of pre-eclampsia can be made in the absence of proteinuria if the new-onset hypertension is associated with thrombocytopenia, impaired liver function, new-onset renal failure, pulmonary oedema or new-onset visual or cerebral disorders, and uteroplacental disorder that results in fetal growth retardation. Bearing in mind that the hypercoagulable state in pre-eclampsia is further enhanced compared to normal pregnancy, the aim of this study was to evaluate the usefulness of global haemostatic assays, endogenous thrombin potential and overall haemostatic potential in the assessment of the haemostatic disorders in pre-eclampsia before and after delivery and to analyse the results of these assays in relation to the outcomes of preeclamptic pregnancy. The characteristics of fibrin clots in normal and preeclamptic pregnancies were also analyzed. Furthermore, different phenotypes of extracellular vesicles were determined and their association with the investigated hemostatic parameters was analyzed. The study involved 46 pregnant women with pre-eclampsia and 80 healthy pregnant women..

Phosphatidylserine Exposing Extracellular Vesicles in Pre-eclamptic Patients

Background: Pre-eclampsia (P-EC) is associated with systemic inflammation, endothelial dysfunction and hypercoagulability. The role of extracellular vesicles (EVs) in coagulation disturbances affecting the development and severity of P-EC remains elusive. We aimed to evaluate the concentration of EVs expressing phosphatidylserine (PS) and specific markers in relation to the thrombin and fibrin formation as well as fibrin clot properties, in pregnant women with P-EC in comparison to healthy pregnant women of similar gestational age. Methods: Blood samples of 30 pregnant women diagnosed with P-EC were collected on the morning following admission to hospital and after delivery (mean duration 5 days). The concentration of the PS-exposing EVs (PS+ EVs) from platelets (CD42a+, endothelial cells (CD62E+), and PS+ EVs expressing tissue factor (TF) and vascular cell adhesion molecule 1 (VCAM-1) were measured by flow cytometry. Further phenotyping of EVs also included expression of PlGF. Markers of maternal haemostasis were correlated with EVs concentration in plasma. Results: Preeclamptic pregnancy was associated with significantly higher plasma levels of PS+ CD42a+ EVs and PS+ VCAM-1+ EVs in comparison with normotensive pregnancy. P-EC patients after delivery had markedly elevated concentration of PS+ CD42a+ EVs, CD62E+ EVs, TF+ EVs, and VCAM-1+ EVs compared to those before delivery. Inverse correlation was observed between EVs concentrations (PS+, PS+ TF+, and PlGF+) and parameters of overall haemostatic potential (OHP) and fibrin formation, while PS+ VCAM-1+ EVs directly correlated with FVIII activity in plasma. Conclusion: Increased levels of PS+ EVs subpopulations in P-EC and their association with global haemostatic parameters, as well as with fibrin clot properties may suggest EVs involvement in intravascular fibrin deposition leading to subsequent microcirculation disorders