Platelet activation and endogenous thrombin potential in pre-eclampsia

Background: Microparticles may be generated from a number of cell types and are known to play a role in haemostasis by a variety of mechanisms. We investigated the role of platelet, red cell, and leucocyte-derived microparticles in the measurement of thrombin generation.Methods: Four parameters of thrombin generation (the endogenous thrombin potential (ETP), lag time, time to peak, peak height) and microparticle content was determined in 35 plasma samples from normal individuals pre and post filtration to remove microparticles. Immunofluorescent flow cytometry was used to identify and enumerate platelet, leucocyte, monocyte and red cell derived microparticles in plasma samples based on the expression of CD42b, CD45, CD15, and Glycophorin A respectively. Expression of phosphatidylserine and tissue factor by microparticles was determined by Annexin V and anti CD142 binding. The pre and post filtration results were compared.Results: There was a significant decrease in ETP and Peak Height, and an increase in the time to peak post filtration (P < 0.001). A significant decrease in the number of CD421, CD451, CD151, CD1421, and Annexin V1 microparticles was also observed. The change in CD42b1 microparticles correlated highly with the change in Annexin V1 microparticles (r 5 0.68). Whilst the change in ETP correlated best with the change in CD151 microparticles (r 5 0.45) and the change in time to peak correlated with the change in Annexin V binding (r 5 0.52) (P < 0.01).Conclusion: The presence of micropartcles in plasma significantly affects thrombin generation. V C 2010

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“…Both calibrator and thrombin generation wells were measured in triplicate. Fluorescence was measured at 390/460 nm every 30 s for 40 min (11).…”

Section: Thrombin Generation Assaymentioning

confidence: 99%

“…Beads 1.1 lm (1 ll) from a manufacturers stock solution diluted 1:1,000 in filtered TS were added to each sample together with Accucount beads (25 ll). Samples were diluted to 1 ml with filtered TS and analysed immediately by flow cytometry as described previously (11)(12)(13).…”

Section: Immunolabellingmentioning

confidence: 99%

Flow cytometric analysis of microparticle phenotype and their role in thrombin generation

2010

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“…This assay is known to define the dynamics and intensity of thrombin development; thrombin is the key hemostatic enzyme and relates to the group of integral indexes of the coagulation system (Hemker et al, 2000;Hemker et al, 2003;Hemker et al, 2006;Regnault et al, 2003;Wielders et al, 1997). This methodological approach has been successfully tested as a part of the complex estimation of the hemostasis system during pregnancy (Dargaud et al, 2010), preeclampsia (Macey et al, 2010) and oral contraceptive intake (Tchaikovski et al, 2007). Recent publication presented interesting data on the specifics of thrombin generation in blood plasma within IVF cycle (Westerlund et al, 2012).…”

Section: Data On the Ivf Results As Affected By Pathologies Of Hemostmentioning

confidence: 99%

The Means of Progress in Improving the Results of in vitro Fertilization Based on the Identification and Correction of the Pathology of Hemostasis

Момот¹,

Lydina²,

Цывкина³

et al. 2012

Enhancing Success of Assisted Reproduction

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“…34 Ayrıca sağlıklı kadınlar, sağlıklı gebeler ve preeklamptik gebelerden oluşan gruplarda TGA testinin incelendiği bir ça-lışmada, ETP değeri, preeklamptik gebelerde sağ-lıklı gebelere göre ve sağlıklı gebelerde gebe olmayan kadınlara göre istatistiksel olarak anlamlı düzeyde daha yüksek bulunmuştur. 35 Benzer olarak preeklampsi öyküsü olan kadınlar ile sağlıklı kadınlarda TGA testinin incelendiği bir araştır-mada, preeklampsi öyküsü olan kadınlarda gebelikten ortalama 15 ay sonra trombin üretimi yüksek olarak saptanmıştır. 21 Bizim çalışmamızda da özellikle trombofili pozitif olan hasta grubunda TGA testi, ETP değeri sağlıklı kontrol grubuna göre daha yüksek bulunmuştur.…”

Section: Discussionunclassified

“…[39][40][41][42] Macey ve ark.nın preeklamptik gebeler, sağ-lıklı gebeler ve sağlıklı gebe olmayan kadınlarda yaptığı çalışmada ise duraklama zamanı ve pik zamanı preeklamptik gebelerde preeklampsisi olmayan gebelere göre, preeklampsisi olamayan gebelerde ise sağlıklı kadınlara göre daha kısa bulunmuştur. 35 Bu çalışmada hastaların trombofili göstergeleri hakkında veri bulunmamaktadır. Farklı protrombotik durumlarda TGA testinin ETP değeri dışındaki parametrelerinin değişimleri hakkında daha fazla çalışmaya ihtiyaç vardır.…”

Section: Discussionunclassified