Does plateletpheresis induce a hypercoagulable state? A global assessment of donor's hemostatic system by ROTEM.

Since there is still debate on the effects of plateletpheresis on coagulation system, we aimed to perform a global assessment of donor's hemostatic function undergoing plateletpheresis by rotation thromboelastometry (ROTEM) analysis and to clarify if plateletpheresis procedure induces a hypercoagulable state. Thirty male plateletpheresis donors were included in the study. Four blood samples were drawn at different time intervals: before the beginning of the apheresis procedure; immediately after the completion of the apheresis procedure; 24 h and 7 days after the apheresis procedure. "Hypercoagulability" was diagnosed readily by having an accelerated clot formation, as evidenced by shortening of CFT and an increase of the clot strength, as evidenced by increasing of MCF. In INTEM assay, CFT value after apheresis was significantly prolonged compared with baseline value while CFT value 7 days after apheresis was significantly shortened compared with values immediately and 24 h after apheresis (p < 0.001). However, CFT-INTEM still did not show any shortening in any of the measurements when compared to pre-apheresis value. MCF value after apheresis was significantly shortened compared with baseline value while MCF value 7 days after apheresis was significantly prolonged compared with values immediately and 24 h after apheresis (p < 0.001). However, MCF-INTEM still did not show any increase in any of the measurements when compared to pre-apheresis value. There was no significant difference in CT value between four measurements (p = 0.064). In EXTEM assay, CFT value after apheresis was significantly prolonged compared with baseline value while CFT value 7 days after apheresis was significantly shortened compared with values immediately and 24 h after apheresis (p < 0.001). However, CFT-EXTEM still did not show any shortening in any of the measurements when compared to pre-apheresis value. MCF values immediately and 24 h after apheresis were significantly shortened compared with baseline value while MCF value 7 days after apheresis was significantly prolonged compared with values immediately and 24 h after apheresis (p < 0.001). However, MCF-EXTEM still did not show any increase in any of the measurements when compared to pre-apheresis value. We found no differences in CT value between four measurements (p = 0.208). Since ROTEM tracings on both INTEM and EXTEM assays did not reveal any significant shortening of CFT and increasing of MCF in any of the measurements after apheresis procedure, we concluded that plateletpheresis does not induce a hypercoagulable state in healthy donors.

Thromboelastometry profile in children with beta-thalassemia.

Beta (ß)-thalassemia is characterized by a hypercoagulable state and an increased risk of thrombosis, which can result in significant morbidity and mortality. The coagulation pattern and determinants of thrombosis in patients with ß-thalassemia remain largely unknown. The aim of this study was to evaluate the whole blood thromboelastometry (TEM) profile of ß-thalassemic children by ROTEM(®). ROTEM(®) assays (INTEM, EXTEM) and traditional coagulation parameters (platelet count, prothrombin time, activated partial thromboplastin time, and fibrinogen) were performed on blood samples from 17 subjects with ß-thalassemia and 19 non-thalassemic controls. Maximum clot firmness (MCF) was significantly higher in subjects with ß-thalassemia than in controls on EXTEM and INTEM analysis (p < 0.001 and p < 0.001, respectively). Of the patients with ß-thalassemia, MCF was higher and clot formation time was shorter in splenectomized subjects than in non-splenectomized subjects on EXTEM and INTEM (p = 0.026, p = 0.002, p < 0.001, p < 0.001, respectively). TEM profiles in ß-thalassemic children were more hypercoagulable compared with controls. Larger prospective studies are needed to evaluate the relevance of the association between ROTEM(®) profile and thromboembolic events in patients with ß-thalassemia.

The role of hemostatic mechanisms in the development of thrombosis in Behcet's disease: an analysis by modified rotation thromboelastogram (ROTEM).

Behcet's disease (BD) is a multisystemic disorder characterised by recurrent oral and genital ulcers. Vasculitis and thrombotic events are the most important causes of mortality. Thrombosis is the major clinical finding in patients with BD, but the cause of the thrombosis is still unclear. Thromboelastography is an alternative method to evaluate almost all steps of the hemostatic system. Today, the modified rotation thromboelastogram (ROTEM) is a newer coagulation test and a more powerful technique. Our aim in this study was to investigate whether hemostatic mechanisms play a role in the development of thrombosis in BD patients by using ROTEM. Thirty BD patients, 20 ankylosing spondylitis patients, and 14 healthy controls who are not taking anti-aggregant or anti-coagulant therapy were included in the study. Whole blood count, protrombin time, activated protrombin time, fibrinogen, D-dimer levels, and ROTEM parameters (clotting time, clot formation time (CFT), and maximum clot formation(MCF)) were determined by INTEM and EXTEM analysis. Of the 30 patients with BD, 19 were women and 11 were men, and mean age was 40.6 ± 11.2. Two of the BD patients had vascular involvement, but none of them were in active phase of the disease during the study. In INTEM assay, MCF (p < 0,001) value was significantly increased, and CFT (p>0.05) value was decreased in BD patients compared with the control group. In the EXTEM assay, there was a similar significant increase in MCF (p=0.002) value and a decrease in CFT (p=0.002) value in BD patients compared with the control group. The results of our study indicated that primary hemostatic mechanisms which can be detected by ROTEM may play a role in the development of thrombosis in patients with BD.

Evaluating coagulation disorders in the use of bevacizumab for metastatic colorectal cancer by thrombelastography.

The aim of this study was to evaluate coagulation disorders in patients with metastatic colorectal cancer treated with bevacizumab by using rotation thrombelastogram (ROTEM(®)) and correlate ROTEM(®) parameters with routine coagulation tests. A total of 18 colorectal cancer patients who received bevacizumab combined with chemotherapy were included. There was no statistically significant difference between results of platelet count, prothrombin time (PT) and activated partial thromboplastin time (APTT), fibrinogen, and D-Dimer obtained at baseline and on day 1 of chemotherapy cycles 4, 8, and 12. CFT value was significantly increased on day 1 of cycle 12 compared with baseline value by both INTEM and EXTEM assays while CT and MCF showed no significant difference. Correlation analysis revealed significant correlation between laboratory parameters and ROTEM(®) parameters. Platelet count showed a positive correlation with MCF in INTEM (r = 0.627) and EXTEM (r = 0.699) assays while showed a negative correlation with CFT in EXTEM (r = -603). There was a significant negative correlation between fibrinogen levels and CFT in INTEM (r = -0.617) and EXTEM (r = -0.512). Our data demonstrated the value of TEG over conventional coagulation tests in evaluating antiangiogenesis agents-induced coagulation disorders.

Aspirin-resistance frequency: a prospective study in 280 healthy Turkish volunteers.

This study reports the frequency of aspirin resistance and its correlation with clinical and biochemical parameters among 280 healthy Turkish volunteers (179 men, 101 women) who were taking 100 mg of aspirin 7 days or more. Aspirin resistance was detected by optical platelet aggregometry, using adenosine diphosphate and arachidonic acid, and defined as a mean aggregation of 64% or more with 5AmicroM adenosine diphosphate and a mean aggregation of 20% or more with 0.5-mg/mL arachidonic acid. Of the study population, 27.5% (26 women [25.5 %] and 51 men [28.5 %]) were aspirin resistant. The current findings indicate that aspirin resistance is an important and real laboratory diagnosis given its frequency of 27.5% in the study population. These results of this large trial evaluating the frequency of aspirin resistance in healthy subjects indicate that aspirin resistance should be diagnosed so that individuals with no response can receive alternative or additional antiplatelet therapy.

Laboratory investigation of hypercoagulability in cancer patients using rotation thrombelastography.

The goal of this study was laboratory testing for hypercoagulability in patients with solid tumors using rotation thrombelastogram (ROTEM) and correlate ROTEM parameters with routine coagulation tests. A total of 78 untreated patients with cancer were included: 28 gastrointestinal system tumors (group 1), 27 respiratory system tumors (group 2), and 23 miscellaneus group of ovarian, renal, nasopharyngeal, mesothelioma, and unknown origin (group 3). Platelet count was significantly increased in group 2 in respect to group 3 (P < 0.05) and fibrinogen level was significantly increased in group 2 in respect to group 1 (P < 0.05). There was no statistically significant difference between subgroups in respect to TEG parameters. Tumor-node-metastasis (TNM) stages of patients was not also associated with either of TEG parameters. Correlation analysis revealed significant correlation between laboratory parameters and ROTEM parameters. Fibrinogen showed the strongest correlation with MCF (r > 0.7) and CFT in all assays (INTEM, EXTEM, FIBTEM, APTEM). There were also statistically significant correlations between platelet number and other ROTEM parameters (INTEM-CFT, -MCF, EXTEM-CFT, -MCF, FIBTEM-MCF, APTEM-CFT, -MCF). In conclusions, our data demonstrates thromboelastographic signs of hypercoagulability in patients with solid tumors. ROTEM is able to identify the contribution of fibrinogen and platelets to clot strength in this patient population.

Prognostic impact of chromosome alterations detected by FISH in Turkish patients with B-cell chronic lymphocytic leukemia.

The goal of this study was to evaluate the relation of chromosomal abnormalities detected by fluorescence in situ hybridization (FISH) in the prognosis of B-cell chronic lymphocytic leukemia (B-CLL) patients. We evaluated the common recurrent chromosomal aberrations in 79 B-CLL patients (51 men, 28 women; mean age 64.3+/-1.2) by FISH analysis using 11q22.3 (ATM), 13q14.3 (13S319 and 13S25), CEP12, and 17p13.1 (TP53) specific probes. Of the 79 patients analyzed by FISH, 40 or 50.6% had at least one aberration. In particular, 34 (43%) patients had a single abnormality and 6 (7.6%) patients had 2 abnormalities. The most frequent abnormality was 13q14.3 deletion, which was detected in 26 (32.9%) patients. Trisomy 12 was seen in 12 (15.2%) cases, and was followed by 17p13.1 (TP53) deletions and 11q22.3 (ATM) deletions in 6 (7.6%) and 4 (5.1%) patients, respectively. When the overall frequencies of these chromosomal aberrations were distributed according to RAI stages, the majority of patients with 13q14.3 deletion (55%), trisomy 12 (70%), and ATM or TP53 deletions (66.7 %) were in advanced stages of disease (RAI II-IV). The overall survival durations in good, intermediate, and poor prognostic groups were 51+/-1.3, 50.9+/-8.6, and 12+/-3.3 months, respectively. Our data suggests that FISH analysis of B-CLL patients provides important diagnostic, clinical, and prognostic information which may help clinicians assess the prognosis and make appropriate treatment decisions.

Platelet function testing during 5-day storage of single and random donor plateletpheresis.

Platelet concentrates are routinely manufactured from whole blood by differential centrifugation (random donor platelets-RDP) or by plateletpheresis (single donor platelets-SDP). These platelet concentrates have a storage period of 5 days and many different approaches exist to measure the condition of platelets during their storage. In this study, platelet aggregation testing using adenosine diphosphate (ADP) and collagen and flow cytometric platelet activation analysis using CD41 FITC and CD62 PE before and after ADP was performed on days 1, 3 and 5 of storage of platelet preparations. Thirty three RDPs, stored in Baxter and Kansuk blood bags and 18 SDPs stored in Fresenius blood bags were evaluated. In RDPs and in SDPs; ADP and collagen induced PA responses were decreased significantly on the 3rd and 5th days compared to 1st day. CD62 positive platelet percentage after ADP were decreased significantly on the 3rd and 5th days compared to the 1st day in Kansuk bags. Flow cytometric analysis revealed minor changes in CD41 expression after ADP on the 3rd day compared to 1st day and on the 5th day compared to 3rd day. Differences in CD62 positive platelet percentage were not significant between the RDPs and SDPs. Our results suggest that: (1) ADP and collagen induced PA responses decrease both in RDPs and SDPs during storage. (2) Flow cytometric analysis does not show major significant changes in platelet activation after ADP during storage. (3) Continous shaking on the agitator does not cause a significant change in CD62 positive platelet percentage during storage. (4) Platelet aggregation responses in RDPs stored in Baxter and Kansuk blood bags do not differ during storage.

Evaluation of inherited and acquired platelet function disorders in iron deficient women with menorrhagia by whole blood lumi-aggregometer.

The commonest cause of iron deficiency anemia (IDA) in premenopausal women is often menstrual blood loss. However, no organic pathology is identified in more than 50% of menorrhagic women. We therefore investigated inherited and acquired bleeding disorders among women with unexplained menorrhagia who developed IDA. In vitro whole blood platelet aggregation (PA) with ADP, arachidonic acid (AA), ristocetin and collagen was studied in addition to full blood count, serum iron levels, serum iron binding capacity, transferrin saturation, ferritin, prothrombin and activated partial thromboplastin time, fibrinogen, D-Dimer, Factor VIII, Factor IX, Factor XI, ristocetin cofactor activity, blood type and bleeding time in 67 women before and after therapy. Before therapy; decreased agonist induced PA was observed in 20% of women by ADP, in 12% by AA, in 2% by ristocetin and in 6% by collagen. After oral iron therapy, decreased platelet aggregation was shown in 8% of women with ADP and 2% of women with AA while initial abnormal ristocetin and collagen induced platelet aggregation responses became normal. Also there was a statistically significant increase of ristocetin cofactor activities and FXI levels after iron repletion. We conclude that; rather than von Willebrand disease, platelet function abnormalities and FXI deficiency are the most common hemostatic disorders in women with unexplained menorrhagia and significant portion of these disorders can be reversed by iron therapy.

C-Reactive Protein in the Follow-up and Estimation of Infections in Acute Leukemic Patients.

Fifty-five febrile neutropenic episodes were monitored by C-reactive protein (CRP) in 26 patients with acute leukemia. In nonfebrile period of patients, serum CRP level was 0.89 mg/dL (range 0.1-8.8 mg/dL) while it was found to be raised to 9.57 mg/dL (range 0.5-24.1 mg/dL) in the febrile group (p= 0.0001). Serum CRP level at the onset of fever was 9.25 mg/dL (range 0.1-16.2 mg/dL) in patients in whom fever was treated succesfully with antipseudomonal beta-lactam antibiotic and aminoglycoside therapy declined to normal levels (p= 0.01); 17.0 mg/dL (range 5.2-33.5 mg/dL) in patients still persisting fever with this combined therapy and so obtained vancomycin (p= 0.001); 16.6 mg/dL (range 0-38.7 mg/dL) in patients required amphotericin B in addition (p= 0.001). In the initial febrile episode; fever resolved at day 3.3 ± 1.21 in patients with serum CRP value below 5 mg/dL; at day 4.42 ± 1.88 in patients with serum CRP value between 10-15 mg/dL; at day 5.14 ± 1.68 in patients with serum CRP value above 15 mg/dL. There was no statistical difference at the time of fever resolution among the first three groups. There was a remarkable difference between groups with CRP values below 5 mg/dL and above 15 mg/dL (p= 0.04). We conclude that determination of serum CRP level before and after infection in febrile neutropenic patients is useful for the follow-up and estimation of febrile neutropenic episode in acute leukemic patients.

Langerhans cell histiocytosis with transformation to acute leukemia showing 45,X, t(8; 21), 5q-, -Y karyotype.

A 31 -year-old man was admitted to hospital with onset of difficulty in walking and urinary incontinence, leading to the diagnosis of Langerhans cell histiocytosis (LCH) which was replacing a thoracic vertebra. Four months after the completion of radiation therapy, he was referred to our department with persistent fever and severe pyogenic ulceration mainly affecting the right-hip. A diagnosis of acute non-lymphoblastic leukemia (ANLL) was made. Cytogenetic studies showed 45,X, t(8; 21), 5q-, -Y We report this case because, development of acute leukemia after LCH is rare and the literature searched for any cytogenetic study in these kind of cases yielded no data.