Multiple Electrode Whole Blood Aggregometry, PFA-100, and In Vivo Bleeding Time for the Point-of-Care Assessment of Aspirin-Induced Platelet Dysfunction in the Preoperative Setting

2011 ◽  
Vol 113 (1) ◽  
pp. 31-39 ◽  
Author(s):  
Csilla Jámbor ◽  
Klaus-Werner von Pape ◽  
Michael Spannagl ◽  
Wulf Dietrich ◽  
Andreas Giebl ◽  
...  
Keyword(s):  
Whole Blood ◽  
Bleeding Time ◽  
Point Of Care ◽  
Platelet Dysfunction ◽  
Care Assessment ◽  
Whole Blood Aggregometry ◽  
Multiple Electrode

Whole Blood Multiple Electrode Aggregometry Is a Reliable Point-of-Care Test of Aspirin-Induced Platelet Dysfunction

2009 ◽  
Vol 109 (1) ◽  
pp. 25-31 ◽  
Author(s):  
Csilla Jámbor ◽  
Christian F. Weber ◽  
Konstanze Gerhardt ◽  
Wulf Dietrich ◽  
Michael Spannagl ◽  
...  
Keyword(s):  
Whole Blood ◽  
Point Of Care ◽  
Platelet Dysfunction ◽  
Multiple Electrode Aggregometry ◽  
Point Of Care Test ◽  
Multiple Electrode

scholarly journals The Role of Whole Blood Platelet Aggregation Studies in the Diagnosis of Unexplained Bleeding Tendencies

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 2260-2260
Author(s):  
Nicole De Simone ◽  
Ravi Sarode ◽  
Sean Yates ◽  
Karen Matevosyan ◽  
Manasa Reddy ◽  
...  
Keyword(s):  
Arachidonic Acid ◽  
Platelet Aggregation ◽  
Platelet Function ◽  
Whole Blood ◽  
Selective Serotonin ◽  
Impedance Method ◽  
Platelet Dysfunction ◽  
Atp Secretion ◽  
Abnormal Results

Abstract Introduction: Platelet aggregation studies (PAS) are an important and underutilized diagnostic test (due to non-availability in most clinical laboratories and the requirement to be performed within 4 hours of sample collection) used in the evaluation of unexplained mucocutaneous type of bleeding after ruling out von Willebrand disease. Platelet aggregation studies are typically performed by one of two methods: impedance method using whole blood aggregometry (WBA) and light transmission aggregometry (LTA) using platelet rich plasma (PRP). WBA confers several advantages over LTA. First, it does not require centrifugation, which not only reduces testing time by half, but also avoids platelet activation and loss of giant thrombocytes. Second, in vivo conditions are better replicated reflecting the natural milieu including red and white blood cells, which are known to affect platelet function in vivo. In addition, WBA requires smaller blood volume making testing feasible for neonates and pediatric patients. Lastly, simultaneous assessment of platelet ATP release is performed to assess secretion defects. Despite these advantages, WBA is not commonly used. Aims: To analyze our data to further support the diagnostic utility of WBA in identifying platelet dysfunction as the etiology of bleeding tendencies. Methods: A retrospective chart review of patients on whom PAS were performed between June 2011 and September 2014. Results: We performed 202 PAS on 162 patients. 82% of patients were females and the average age was 28 years (range 9 months-87 years). 24 (15%) patients were pediatric (range 9 months-18 years). 83 of 162 (51%) patients had abnormal results (52% of adults and 50% of the pediatric cases). 26 of the 162 (16%) patients had repeat studies performed. Of these patients, 77% (20/26) had reproducible findings that confirmed the previous results. 8% (2/26) had normalized platelet function after discontinuation of medications (e.g. statins, fish oil, selective serotonin reuptake inhibitor) known to induce platelet dysfunction. 15% (4/26) had different responses to agonists on repeat testing. Abnormal WBA studies revealed decreased to absent responses to various agonists described in table 1. In patients on selective serotonin release inhibitors (SSRIs), there was a spectrum of responses to agonists; the most common abnormality was global dysfunction. Abnormalities to single agonists, such as ADP and AA, were also seen in patients taking SSRIs. Non-steroidal anti-inflammatory drugs affected aggregation with arachidonic acid (AA) and AA+ADP. Statins affected aggregation with AA alone, AA+ADP and AA+ATP secretion. 3 patients had platelet dysfunction consistent with Acquired Glanzmann's Syndrome due possibly to autoantibodies in the setting of chronic lymphocytic leukemia. Conclusion: Over 50% patients tested by WBA had abnormal platelet function giving high positive predictive value for this test in a selected group of patients who otherwise would have carried a non-specific bleeding diagnosis with non-specific treatment. Table 1. Distribution of Agonists Eliciting Impaired Responses Agonists Eliciting Impaired Response Number of Studies with Abnormal Results AA+Collagen (Aspirin like defect) 27 (23%) AA+Collagen+ADP 22 (18%) AA+ADP 21 (17%) AA+Collagen+ADP+Ristocetin (Global dysfunction) 19 (15%) ADP 11 (9%) AA 7 (6%) ADP+Collagen 4 (3%) AA+ADP+Ristocetin 3 (2%) Decreased ATP Secretion 8 (7%) AA=Arachidonic Acid Disclosures No relevant conflicts of interest to declare.

scholarly journals PENGARUH EKSTRAK ETANOL RIMPANG KUNYIT (Curcuma Longa) TERHADAP PEMERIKSAAN HEMOSTASIS DARAH

2021 ◽  
pp. 280-292
Author(s):  
Rini Roslaeni ◽  
Raya Agung Maha Sakti ◽  
Mochamad Fathan Zulfahmi ◽  
Maharany Rizky Yuniar ◽  
Evi Sovia ◽  
...  
Keyword(s):  
Prothrombin Time ◽  
Whole Blood ◽  
Bleeding Time ◽  
Curcuma Longa ◽  
Per Oral

Kunyit (Curcuma longa) merupakan tanaman obat yang banyak terdapat di Asia, sering digunakan sebagai bumbu ataupun obat tradiosional. Tanaman kunyit memiliki akar (rimpang) berwarna kuning tua dan mengandung senyawa penting, diantaranya curcumin dan ar-tumeron yang berperan dalam menghambat pembekuan darah. Saat ini salah satu penyebab utama morbiditas dan mortalitas di dunia adalah penyakit kardiovaskular seperti stroke dan penyakit jantung koroner. Potensi kunyit yang dapat bersifat antikoagulan diharapkan menjadi alternatif pencegahan penyakit kardiovaskular. Tujuan penelitian untuk mengetahui efek ekstrak etanol rimpang kunyit secara in vivo terhadap bleeding time (BT), dan secara in vitro terhadap prothrombin time (PT) dan retraksi bekuan. Uji in vitro menggunakan sampel darah manusia yang ditambahkan ekstrak rimpang kunyit, sedangkan uji in vivo dengan cara memberikan ekstrak rimpang kunyit secara per oral terhadap mencit. Pemeriksaan PT menggunakan metode tilt tube, pemeriksaan retraksi bekuan menggunakan whole blood lalu didiamkan selama 2 jam dan dihitung sisa serumnya (volume serum/volume darah awal x100%), sedangkan pemeriksaan BT dilakukan dengan menginsisi ekor tikus. Hasil penelitian menunjukkan rerata PT dan BT memanjang pada semua semua kelompok uji. Semakin besar dosis ekstrak rimpang kunyit maka semakin memanjang nilai PT dan BT. Retraksi bekuan pada kelompok uji menunjukkan rerata persentase serum yang rendah dibandingkan kontrol negatif, dan semakin besar dosis ekstrak yang diberikan maka semakin rendah nilai retraksi bekuannya. Hal tersebut diduga karena zat-zat aktif seperti curcumin dan ar-tumeron dapat menghambat proses pembekuan darah sehingga memengaruhi hasil pemeriksaan hemostasis darah.

Multiple Electrode Whole-Blood Aggregometry and Bleeding in Cardiac Surgery Patients Receiving Thienopyridines

2011 ◽  
Vol 91 (1) ◽  
pp. 123-129 ◽  
Author(s):  
Marco Ranucci ◽  
Ekaterina Baryshnikova ◽  
Giorgio Soro ◽  
Andrea Ballotta ◽  
Donatella De Benedetti ◽  
...  
Keyword(s):  
Cardiac Surgery ◽  
Whole Blood ◽  
Whole Blood Aggregometry ◽  
Multiple Electrode

scholarly journals A Novel Point-of-Care Whole Blood Coagulation Assay to Monitor Emicizumab Therapy in Patients with Hemophilia

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 2475-2475 ◽  
Author(s):  
Michael A Suster ◽  
Debnath Maji ◽  
Lalitha V. Nayak ◽  
Christina Jenkins ◽  
Susan Hunter ◽  
...  
Keyword(s):  
Whole Blood ◽  
Research Funding ◽  
Hemophilia A ◽  
Ex Vivo ◽  
Point Of Care ◽  
Blood Samples ◽  
University Patents ◽  
Whole Blood Samples ◽  
Western Reserve

Abstract Introduction: Emicizumab is a humanized, bispecific antibody against activated Factor IX (FIX/IXa) and FX/Xa that mimics the cofactor function of activated FVIII (FVIIIa) by spatially relocating FIXa and FX to the appropriate position in the tenase complex. Though the bleeding rate in patients on emicizumab is remarkably reduced, hemostatic monitoring is important in the event of breakthrough bleeding, development of anti-drug antibodies, and for surgery. An APTT is markedly shortened in the presence of small amounts of emicizumab, and, hence not useful. The thrombin generation assay (TGA), thromboelastometry, and clot waveform analysis measure the hemostatic effect of emicizumab, both in vivo and ex vivo. However, these assays are time-consuming, need expert interpretation, and not widely available. We have developed a novel, point-of-care, whole blood assay based on a dielectric microsensor (ClotChip) that can obtain global blood coagulation assessment in a miniaturized, portable measurement platform. The aim of this study was to assess the sensitivity of ClotChip to detect the addition of variable concentrations of emicizumab ex vivo into hemophilic whole blood and the feasibility of monitoring emicizumab therapy in vivo using ClotChip . Methods: After IRB approval, patients accrued for the study were adults with severe hemophilia A with inhibitors starting emicizumab therapy (n=2), and a child with severe hemophilia A without inhibitors (n=1). Blood samples were obtained by venipuncture into collection tubes containing 3.2% sodium citrate anticoagulant. Samples were collected both prior to (n=2), and at time points of 30 min (n=1), 1 hour (n=1), 1 week 1 (n=1), and 8 weeks (n=1) after initiation of emicizumab therapy. Whole blood samples from hemophilia patients not on emicizumab were spiked with different concentrations of emicizumab, with and without the addition of replacement therapy. Whole blood samples were then tested with the ClotChip. Coagulation was induced with CaCl2 addition. ClotChip is based on the electrical technique of dielectric spectroscopy (DS) integrated into a low-cost (material cost < $1), small- sized (26mm × 9mm × 3mm), and disposable microfluidic biochip with miniscule sample volume (< 10 µL). ClotChip curves were calculated as blood permittivity at 1MHz, and the time to reach a peak in permittivity (TpeakFig 1A) was taken as an indication of coagulation time. TGA using low tissue factor concentration was also performed on blood samples according to the manufacturer's direction. Results: We observed a decrease in the ClotChip Tpeak parameter for post-therapy samples (30 min and 1 hr.) compared to baseline (pre-therapy) for hemophilia patients with inhibitors on emicizumab therapy (Fig 1B). A time-dependent decrease was observed in ClotChipTpeak after emicizumab administration with week 1 and 8 samples showing normal values. Ex vivo spiking with emicizumab in blood from patients with hemophilia with and without inhibitors showed a concentration dependent decrease in ClotChip Tpeak parameter (Fig 1C). Addition of rFVIII or rFIX in emicizumab-spiked blood from the patient without inhibitors further decreased ClotChipTpeak in a concentration dependent manner (Fig 1D). Similarly, addition of rFVIII to the emicizumab spiked blood from the inhibitor patient further decreased ClotChip Tpeak compared to emicizumab alone (Fig 1E). ClotChip Tpeak exhibited strong negative correlation with ETP (rs = 0.81, Fig 1F) and peak thrombin (rs = -0.82) from TGA assays. Conclusions: Our studies demonstrate the feasibility of monitoring emicizumab therapy in patients with hemophilia with and without inhibitors, using a novel, microfluidic, dielectric sensor (ClotChip), allowing whole blood assessment of hemostasis in a single disposable sensor. ClotChip has potential to fulfill an unmet clinical need to assess global coagulation potential in hemophilia patients on emicizumab therapy, especially when additional hemostatic therapy is needed for treatment of breakthrough bleeds. Additional spiking studies to assess the addition of bypassing agents (rFVII or FEIBA) to blood from emicizumab-treated patients are underway. Disclosures Suster: Case Western Reserve University: Patents & Royalties: licensed to XaTek, Inc.; XaTek, Inc.: Consultancy. Maji:Case Western Reserve University: Patents & Royalties: Licensed to XaTek, Inc. . Schmaier:Temple University: Patents & Royalties; Shire: Consultancy, Honoraria, Research Funding; Enzyme Research Laboratories: Honoraria; Cleveland Clinic Foundation: Research Funding; Alnylam: Research Funding; Biomotiv: Consultancy. Mohseni:Case Western Reserve University: Patents & Royalties: Licensed to XaTek, Inc.; XaTek, Inc.: Consultancy. Ahuja:Shire: Honoraria, Speakers Bureau; Bayer: Honoraria; Bioverativ: Honoraria, Speakers Bureau.

Hypoxia Reoxygenation Treatment Induces Platelet Hyperactivity and Relieves Calpain-1-Mediated Inhibition of Platelet Aggregation in a Mouse Model of Severe Sickle Cell Disease

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 413-413
Author(s):  
Jennifer O. Nwankwo ◽  
Rod R. Warburton ◽  
Thomas Gremmel ◽  
Anja J. Gerrits ◽  
Lauren J. Richey ◽  
...  
Keyword(s):  
Steady State ◽  
Platelet Aggregation ◽  
Platelet Activation ◽  
Sickle Cell ◽  
Whole Blood ◽  
Research Funding ◽  
Blood Platelet ◽  
Platelet Dysfunction ◽  
Inhibition Of Platelet Aggregation

Abstract Introduction: With an estimated 15 million patients and no drug that addresses its etiology, sickle cell disease (SCD) remains an area of unmet need. Vaso-occlusive pain crisis (VOPC), the hallmark of SCD, is initiated by sickle RBCs (sRBCs) recruiting leukocytes and platelets to potentiate vessel occlusion. ADP released by sRBCs is a potent activator of platelets, and sickle cell patients are known to have activated platelets in circulation both at steady state and during VOPC. However, the mechanism underlying platelet dysfunction in SCD is not fully understood. Platelet activation mediated by the protease activated receptors (PAR1 and PAR4 in humans, PAR3 and PAR4 in mice), triggers PLC-β activation resulting in calcium mobilization. The increased calcium flux leads to activation of GPIIbIIIa/aIIbb3, GP1b, and P-selectin involved in platelet aggregation, adhesion, and rolling. Prior evidence has established a role of the calcium-activated cysteine protease, calpain-1 in platelet activation. Washed platelets from calpain-1 knockout C57BL/6 mice demonstrated impaired platelet aggregation. However, due to the critical contribution of sRBCs to platelet dysfunction in SCD, whole blood (impedance) aggregometry represents a physiological assessment of platelet aggregation. Methods: Townes SCD mice (SS) were backcrossed with calpain-1 knockout (CKO) mice to generate SCD mice lacking calpain-1 (SSCKO). Humanized mice (AA) were used as controls. Using flow cytometry, we evaluated in vivo platelet activation following stimulation with ADP and Thrombin GPRP. Platelet counts were obtained via ADVIA 120 and flow cytometry. For platelet aggregation, 500 μL of blood was harvested from the vena cava of AA, SS, SSCKO, and CKO mice. Whole blood aggregation in response to PAR4 stimulation was assessed using the Roche Multiplate Analyzer. A separate group of mice were challenged with hypoxia/reoxygenation (H/R) treatment (3 hours of 10.5% O2, followed by 4 hours of 21% O2) prior to platelet aggregation testing. SCD mice are characterized by tissue infarcts suggestive of thrombus formation. To examine whether H/R treatment induces formation of fibrin thrombi, we harvested brain, lungs, heart, kidneys, liver, and spleen following blood collection, and performed histology. Results: Compared to AA, SS and SSCKO mice are thrombocytopenic. Similar to Berkeley, Townes mouse platelets are activated in vivo, demonstrated by activated GPIIbIIIa on circulating platelets. At steady state, PAR4 agonist-induced platelet aggregation is similar in AA and SS mice (64 U v. 53 U, n = 6-10/group, p = 0.3). As depicted in Fig 1., SSCKO mice show significantly reduced platelet aggregation compared to SS mice (13 U v 53 U, p<0.001, n = 6-10 per group). As expected, H/R treatment induces platelet hyperactivity in SS, but not AA mice (71 U v. 59 U, p = 0.04, n = 6/group). Interestingly, the H/R-induced platelet hyperactivity partially relieves the calpain-1-mediated inhibition of platelet aggregation in SSCKO mice at steady state (Fig 2) (53 U v. 13 U, p<0.001, n = 6-8/group). Furthermore, CKO mice do not show defective platelet aggregation at both steady state, and following H/R treatment, indicating an important role for sRBCs and adhesion in whole blood aggregation. Despite attenuated platelet aggregation, SSCKO platelets at steady state show comparable expression of activated GPIIbIIIa relative to their AA and SS counterparts, indicating that the impaired aggregation at steady state is not due to defective integrin translocation. Since the Roche Multiplate Analyzer takes platelet adhesion into account in determining aggregation, we hypothesize that defective SSCKO whole blood aggregation at steady state is due to reduced cell adhesion. Finally, while silent infarcts were detected in all tissues, fibrin thrombi were not detected, a finding consistent with studies in the Berkeley model reporting minimal thrombi formation in tissues. In summary, we report that calpain-1 is required for platelet aggregation in steady state SCD but not following H/R treatment. This is the first study of H/R-induced platelet dysfunction in the Townes model, and raises the possibility of targeting calpain-1 as a treatment for platelet hyperactivity in SCD. Figure 1. Impaired steady state whole blood platelet aggregation in SSCKO mice (panel 3). Figure 1. Impaired steady state whole blood platelet aggregation in SSCKO mice (panel 3). Figure 2. H/R relieves calpain-1-mediated inhibition of platelet aggregation in SS-CKO mice (panel 3). Figure 2. H/R relieves calpain-1-mediated inhibition of platelet aggregation in SS-CKO mice (panel 3). Disclosures Jakubowski: Eli Lilly and Company: Employment, Equity Ownership. Frelinger:GL Synthesis: Research Funding; Celerion: Research Funding; Megakaryon: Research Funding; Bristol-Myers Squibb: Research Funding; Sysmex: Research Funding; Eisai: Research Funding; Baxalta: Research Funding; Pfizer: Research Funding; GE Global Research: Research Funding; NIH: Research Funding.

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