O. Meltem Akay

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.

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

Behcets 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.

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 5µM 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.

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), 5 q -, - 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.

Platelet dysfunction and other hemostatic disorders in women with menorrhagia: the utility of whole blood lumi-aggregometer

Menorrhagia is a very common clinical problem among women of reproductive age and is unexplained in more than 50% of cases [1]. The widely accepted clinical definition of menorrhagia is blood loss of 80 ml, or more, per period [2]. Underlying bleeding disorders, such as von Willebrand disease (VWD) and platelet abnormalities may present as menorrhagia. However, the relevance of a systemic screening for hemostasis in women with menorrhagia remains controversial [3]. Here, we describe the findings of a prospective study investigating the frequency of platelet dysfunction and other hemostatic defects among women with unexplained menorrhagia using whole blood lumi-aggregometer. Sixty-seven Turkish white women aged between 17 and 50 years, with a physician diagnosis of menorrhagia were screened after the study was approved by the local ethics committee. Women with known bleeding or other systemic disorders such as renal, hepatic and endocrine diseases were excluded initially. All cases were examined at the gynecology department and had pelvic ultrasonography; those with submucous uterine fibroids, fibroids more than 2 cm in diameter, uterine polyps, ovarian tumors and intrauterine device were excluded. Women were required to have not ingested combined oral contraceptives and other hormonal based therapy at least one cycle prior to sampling. We also attempted to prevent any medication especially non-steroidal anti-inflammatory drugs, aspirin as well as alcohol in the last 10 days prior to the aggregation studies. All subjects were evaluated on days 3–7 of their menstrual cycle to minimize interindividual variation. Sixteen age-matched women with no menorrhagia served as controls. Both patients and controls underwent the following laboratory tests: hemoglobin, platelet count, activated partial thromboplastin time (aPTT), factors (F) VIII, IX, XI, ristocetin cofactor activity (RCof), platelet aggregation and ATP release. Platelet function testing was performed on whole blood platelet lumi-aggregometer (Chronolog Corporation, Model 560-Ca) using luminescence method in diluted blood (1:1 blood normal saline ratio). Citrated blood was collected under light tourniquet through 19 gauge needles into 4.5-ml vacutainers (Becton Dickinson) containing 3.2% trisodium citrate in a 9:1 blood anticoagulant ratio. The citrate tubes were thoroughly mixed by gentle inversion before dispensing 450 ll of citrated whole blood into cuvettes (chronolog No: 367) each containing 450 ll normal saline and a disposable siliconized stir bar. After the tubes were warmed at 37 C and stirred in the incubation wells, 100 ll luciferin (chronolog No: 395 chrono Lume Reagent) was added to the cuvette and the luminesence callibrated 5 min later; the impedance electrode was placed into the cuvette and calibration checked. After agonist addition platelet aggregation and ATP release tracings were measured over 6 min. The agonist used and their final concentrations were; ADP (Chrono Par 384) 5.0 mM, Arachidonic acid (AA) (Chrono Par 390) 0.5 mM, Ristocetin (Chrono Par 396) 1.0 mg/ml, and Collagen (Chrono Par 385) 2 lg/ml. Calculated platelet aggregation (ohms) and ATP release (nmol) normal ranges (as mean ± standard deviation) were 8–21 ohms and 0.4–2.9 nmol for ADP, 13–23 ohms and 0.8–3.5 nmol for AA, 17–28 ohms and 0.5–2.7 nmol for collagen, 5–16 ohms for ristocetin. O. M. Akay (&) Z. Gulbas Department of Hematology, Eskisehir Osmangazi University Medical School, 26480 Eskisehir, Turkey e-mail: melhak@hotmail.com