Background:Cytomegalovirus (CMV) infection influence platelet recovery in bone marrow transplantation patients. Patients with CMV infection, primary, reactivated, or latent, require more platelet unit transfusions, this procedure determines increases the risk of transfusion‐related infections. In literature there are many reports regarding CMV implication in bone marrow recovery secondary transplant procedure.Aims:The association with CMV infection and slowly recovery of bone marrow secondary chemotherapy in AML was rarely reportedMethods:We report 2 cases with delay in platelet recovery after intensive chemotherapy‐ consolidation phase of acute myeloid leukaemia patients.Results:Patients male 46 years and female 55 years were diagnosed with AML 1. Diagnose was put using WHO criteria and was not identified any genetic abnormalities. We started 3+7 regimen with obtaining complete remission status. After bone marrow recovery we started consolidation phase with high doses Cytarabine 2 applications. After second application we observed delay in platelet recovery (more than 1 months). Bone marrow biopsy indicated severe aplasia. Serology for CMV infection was positive for IgG antibodies, PCR for CMV was positive indicated active CMV infection. Patients were referred to infectionist and was started antiviral treatment – Gancyclovir. During treatment patients presented increases of platelet count and absence of platelet unit transfusion, in 1 month patients had G1 trombocytopenia (improving of CTCAE grade from G4 to G1)Summary/Conclusion:The delay in bone marrow recovery after intensive chemotherapy used as treatment of acute myeloid leukemia patient represent a warning to check CMV infection presence. This attitude could avoid complications like infections or/and transfusions related.

Background:Patients with Myeloid Metaplasia with Myelofibrosis (MMM) present at the onset or during evolution thrombotic complications. In the assessment of thrombosis risk are important the presence of JAK2 mutation as well as platelet function. Receptors that define the status of activated platelets (CD62P, CD36) are better expressed in chronic myeloproliferative neoplasms (MPNs) patients with thrombotic complications. Numerous studies have shown a high oxidative status in MPNs patients, a situation that correlates with a higher thrombotic risk.Aims:The aim of our study was to determine if resting membrane potential could be correlated with alterations in expression of platelet receptors and reactive species production (ROS)MethodsThis retrospective study included 35 patients with MMM (16 male and 19 female), median age 61.1 (95% CI of median 60,71‐72) as well as 15 healthy volunteers. The diagnosis of MMM was made according to World Health Organization (WHO) classification of MPNs. We evaluated the flow cytometry markers of platelet adhesion (CD42a, CD42b), aggregation (CD41, CD61) and CD36. Production of reactive species (ROS) was examined using fluorescence method with DCFDA and was assessed by aria under curve (AUC) in serial measurements during 900 sec. The determination of platelet membrane potential was done by fluorescence method using 3,3′‐dipropyl‐2,2′‐thiadicarbocyanine iodide (DiSC3‐5)Results:The ROS production is significantly higher in MMM group: median value of AUC patient group 12780882862,5000 compared with controls 11621365017,5000, p= 0.04. Patients in advanced phase (grade 5 Hackett of splenomegaly) has high level of ROS production compared with patients diagnosed in early stage (grade 1 or 2 Hackett of splenomegaly) median value: 13178014970 vs 12303298585, p = 0.001. There are no differences between groups that was splitted by type of treatment Ruxolitinib/Hydroxiureea or type of mutation diagnosed JAK2V617F/CALR/no mutation. Patients with MMM present higher resting membrane potential (RMP) compared with controls (median value ‐ 63 mV vs–57.5 mV, p = 0.05). The expression of CD36 receptor was higher in MMM group vs controls: median value 27.91 vs 20.91, p = 0.003. Number of microparticle (MP) derivated from platelet, erythrocyte and endothelial cells are no different between patient and control group. We obtain only lower receptor expression on MP surface for CD41/CD61, CD42b and GlyA in patient group compared with control group, p < 0.05. There are significant correlation between resting membrane potential and expression of CD42b (r ‐0.45, p = 0.04) and GlyA receptors (r ‐0.45, p = 0.04).Summary/Conclusion:Patients with MMM especially those with have a higher level of ROS production especially in advanced phase. The hyperpolarisation of cell macrophages membrane could be associated with high ROS production, no reports are regarding platelet membrane. In our study we did not obtain any significant correlation between RMP and ROS production. The expression of CD 36 is higher in MMM patients but was not correlated with ROS production. It was proved that generation of intracellular ROS is associated with CD36 signaling and represent one start point for thrombosis. Low expression of CD42b and GlyA of microparticle surface is associated with lower RMP that correspond to activated status of platelet membrane. We have to verify these results in higher number of patients in order to find significant correlations

Background: Patients with acute myeloid leukemia older than 60 years old have a poor prognosis and frequently are not fit for standard chemotherapy. Aims: We retrospectively analysed diagnosis characteristics and treatment outcome in order to identify particularities present in elderly acute myeloid leukemia (AML) group. Methods: We enrolled 62 AML patients older than 60 years diagnosed in Hematology Department Colentina Clinical Hospital between 2015-2019. The diagnosis of AML was made according to French-American-British Cooperative Group (FAB) classification combined with the 2008 revision of the World Health Organization (WHO) classification of myeloid neoplasms and acute leukemia. Results: The median age was 70 years (range 72 to 76 years), and the male:female ratio was 1:1.The most frequent AML subtype was AML1 – 29 cases (46,77%) and in many cases AML was secondary myelodisplastic syndrome evolution – 38 cases (61%) Analysis of evolution data revealed an AML subgroup, consisting of 28 newly diagnosed patients, with a particular short survival – less than 2 months. Eleven patients from this subgroup were not treated due infection presence at diagnosis, this complication has negative influence of evolution of AML patients. In 17 patients we started treatment with hypomethylating agents (Azacytidina or Decytabina) and in 3 patients Mercaptopurine. The treatment outcome analysis identified 16 AML patients with unfavourable evolution due infection associated with severe neutropenia; 1 patient was lost due disease progression. Molecular diagnosis revealed FLT3-ITD mutation in 4 patients, 3 of them being lost immediately after diagnosis (1-2 moths). Cytogenetic results were available for 35 AML patients. The majority of patients had complex karyotype changes or other adverse or intermediate risk cytogenetics (25 out of 35 patients). The analyses of survival rate was performed by Kaplan Meier statistic method and used as endpoint the death and as analyses factors: the progression risk, treatment, presence of pancytopenia, infection. We obtained significant difference in patients group older than 75 years old who have infection – median survival rate 1 months (95%CI 1-8 mo) compared with those without infection 21 months (9-27mo), p = 0.05Summary/Conclusion: Acute myeloid leukemia diagnosed in our patients group was frequently secondary to MDS. Intermediate and high risk cytogenetic abnormalities were detected in a majority of these patients. Infection represent an important unfavourable factor in evolution and treatment outcome of patient older than 75 years

We present the case of a patient diagnosed in SUUB Hematology with adult T-cell leukemia- lymphoma. The particularities of this case are the delay in diagnosis, the apparent onset of disease with severe eye and skin determination in the leukemic phase, and the particular lung involvement in the fi nal stage of the disease.

Abstract Myelodysplastic syndromes (MDS) are a heterogeneous group of clonal hematopoietic stem cell disorders; they are characterized by ineffective hematopoiesis and a predilection to the development of acute myeloid leukemia (AML). For a rapid evaluation of the outcome in myelodysplastic patients non-cytogenetic prognostic scores can be used. Aim: This study proposed to demonstrate that age and gender are important factors in the outcome of the patients diagnosed with myelodysplastic syndrome. Materials and methods: This study was conducted in the Department of Hematology of the Emergency University Hospital Bucharest during October 2008 and October 2012. Results: Male sex and age higher than 60 years are associated with high risk in the studied cases by using the Spanish prognostic score. According to Goasguen score: male sex and age, patients older than 60 years, present characteristics associated with an intermediate risk. Based on the Dusseldorf score, age over 60 years and female gender were associated with pronounced risk in the examined group. By examining the Bournemouth score in our group, we found that age > 60 years correlated with a higher frequency of risk, but no significant differences regarding the sex of patients were observed. Conclusions: We concluded that age > 60 years and male gender are important predisposing factors in the survival. Abbreviations MDS = myelodysplastic syndromes, AML = acute myeloid leukemia, LDH = lactate dehydrogenase, FAB = French-American-British, WHO = World Health Organization, IPSS = International Prognostic Scoring System, ALIP = abnormal localization of immature precursors, WPSS = WHO classification-based prognostic scoring system, FISH = fluorescence in situ hybridization, del = deletion.

Background: The diagnosis and management of the patients with chronic lymphoproliferative diseases have become dependent on immunological criteria. Flow cytometry immunophenotyping is used for rapid and specific diagnosis but there are cases when we are not facing a typical immunophenotype, so there is a constant need to find new markers and new combinations of markers that would allow the improvement and the development of our diagnosis. Aim: Our aim was to evaluate CD 200 expression in different B-cell chronic lymphoproliferative disorders. CD200 is a membrane glycoprotein belonging to the immunoglobulin superfamily and the over-expression of CD200 has been reported in a number of malignancies, including CLL, as well as on cancer stem cells. Methods: We analyzed the CD200 expression in 122 patients diagnosed with chronic lymphoproliferative disorders (100 patients with CLL, 10 patients with splenic marginal zone lymphoma (SMZL), 10 patients with MCL and 2 patients with hairy cell leukemia), in the Department of Hematology of the University Emergency Hospital, Bucharest. We performed immunophenotypical analysis of peripheral blood and bone marrow aspiration on BD FACS Calibur flowcytometer. Results: CD200 was brightly expressed in all 100 CLL patients (100%). In SMZL patients, CD200 was dim positive (40%-60%), in patients with HCL. CD200 was also bright positive (96% and 97%) and in patients with MCL CD200 was negative (1-10%); CD 200 was significantly higher in CLL patients compared with other B-cell chronic lymphoproliferative disorders. We found 14 patients with CD19, CD5 positive population and CD23- , but with high expression of CD 200. Cyclin D1 was negative on bone marrow biopsy in 13/14 of these patients. (1/14 patients were without bone marrow involvement); Conclusions: CD200 has a great impact in diagnosing B- chronic lymphoproliferative disorders, especially when we want to determine the origin of a CD19, CD5 positive population and distinguish between CLL and MCL. CD 23 is a reliable marker in those cases, but, as we showed, CD23 might have a lower specificity than CD200 for CLL. We added CD200 in our panels in order to diagnose chronic lymphoproliferative disorders, not to replace CD 23, but to improve and save time in our diagnostic process. The high expression of CD200 in CLL and HCL could open the option for new- targeted therapy (anti-CD200).

Background: Chronic lymphoproliferative disorders (CLD) are frequently found in patients with hepatitis viral infections, which can lead to changes in pathogenesis. Hepatitis viruses are hepatotrope viruses, potentially lymphotrope and also potentially oncogenic (hepatocellular carcinoma) viruses. HBV and HCV are involved in autoimmune disorders and in the ethiopathogeny of chronic lymphoproliferative disorders. Aim: Detection of immunophenotype changes of malignant lymphocytes in CLD – especially CLL – associated with hepatitis viral infections. Materials and methods: Bone marrow aspirate, peripheral blood samples on EDTA were available for analysis from 58 patients from a follow–up schedule of the Department of Hematology SUUB from March 2008 until June 2009. The patients were diagnosed with chronic lymphoproliferative disorders associated with hepatitis virus B/C/D infections. A group of 28 consecutive unselected patients with CLL who met the diagnostic criteria of the National Cancer Institute–Working Group (NCI NCIWG), and associated hepatitis viral infection (v–CLL) were studied for the expression of several immunophenotypical markers, in comparison to CLL patients without viral infection (control group). Immunophenotyping analysis was performed on a FACS Calibur flowcytometer with a large panel according to EGIL/WHO recommendations. The diagnosis was completed after the histological and immunochemical analysis from tumoral lesions. Results: demographics characteristics – male/female ratio 1/2, average age 64 years. Disease type: 90% B–CLD, 5% T–CLD, 5% Hodgkin's disease. The viral infections: 58,53% HCV, 34,41Z% HBV, 2,43% HBV+HDV, 2,43% HCV+HDV, 2,43% HBV+HCV+HDV. We found in CLL with viral coinfection (v–CLL) cases an elevated expression of B–cell markers – CD19 (Md95/92), CD20 (Md 90/39), CD79b (Md58/31), CD23 (Md67/37). Poor prognosis markers have a higher expression in v–CLL: CD38 (Md49/24), Bcl2 (Md 46/5), cyclin D19 (Md 11/0,5). No change in ZAP–70 expression was observed: Md 59,5/59,1. Discussions: Hepatitis viruses could be involved in the pathogenesis of CLD, but as a trigger for a more aggressive outcome. Higher expression of B–cell markers CD19, CD20 in CLL with viral infection suggests a change to atypical CLL, sustained by elevated expression of known poor prognosis markers bcl–2, cyclin D1 and CD38. Lack of ZAP–70 expression could be explained by a strong correlation with a basic unmutated IgVH status, not related to the viral infection. We found a higher frequency of HCV infection in patients with CLD and especially in CLL patients, which were analyzed extensively for immunophenotypical changes. In the present study, we demonstrated that this CD5+ B cell population with clonal expansion, defining CLL patients, has a different immunophenotype, probably related to the hepatitis viral infection.

I. Voican, Universitary Emergency Hospital, Bucharest ABSTRACT Among all BCR-ABL negative chronic myeloproliferative diseases, Polycythemia Vera and Essential Thrombocythemia are most frequently associated with thrombotic events, o en with unusual locations. Hereby we present the case of a patient concomitantly diagnosed with suprahepatic veins thrombosis and PV, where the clinical-pathological picture corroborated with paraclinical data raised the problem of a diff erential diagnosis with Mosse Syndrome. Rapid initiation of an intense cytostatic and anticoagulant treatment, together with pathogenic and symptomatic measures regarding hepatic cirrhosis insured a favorable evolution of the disease, with normalization of hematological parameters, disappearance of phenomenon of liver failure and diminution of the thrombotic process.

Purpose. Even though there are no solid data regarding chemotherapy treated leukemia during pregnancy, the results based on short series reports show that the management of such condition can be safely achieved during the second and third trimester. We present three personal cases of pregnant women treated with cytostatic agents, two of them accidentally receiving complete chemotherapy during the entire pregnancy without malformative consequences. First case. A 19 yrs old woman diagnosed with chronic myeloid leukemia who conceived spontaneously and mistook the pregnancy signs for a relapse of the disease. During the pregnancy she continued the treatment, receiving until the fifth month an association of Hydroxyurea and alfa-interferon and afterwards switched to Imatinib until term. She presented at 38–39 weeks and delivered by cesarean section a little girl of 3510 g in a perfect state of health. The blood count of both mother and child were normal. Second case. A similar situation in a young woman with lymphoblastic acute leukemia under treatment with Vincristin, Methotrexat, Purinethol. She presented in advanced spontaneous labour at 33–34 weeks and delivered a little girl of 1700 g without malformative signs and normal blood count. Third case. A 17 years old girl who was diagnosed with acute myeloid leukemia at 29 weeks pregnancy. She received induction chemotherapy with Ara-C, due to the significant bone marrow infiltrate and disease induced disseminated intravascular coagulopathy. She presented premature uterine contractions at 32 weeks and delivered by cesarean section a premature boy of 1750g with Apgar score 8. The infant did not present any malformation (by clinical and ultrasound examination) and the blood count was normal. The studies have shown so far that in the case of chronic myeloid leukemia, the treatment with Imatinib was associated with 50% apparently normal live infants and that chemotherapy for acute leukemia during the second or third trimester may not require termination of pregnancy, because both remission and delivery of a normal infant are likely to be obtained.

Patients with chronic myeloproliferative disorders have frequently thrombosis, especially for JAK2-positive patients. The aim of this study is to identify and correlate abnormalities of platelet aggregation for 14 JAK-positive patients, with thrombosis incidence. Thromboses were present more frequently in the JAK-positive group than the JAK-negative group. Patients with JAK2 genotype have more diminished platelet response for ADP, epinephrine, and collagen than the JAK-negative group and normal response for ristocetin, but these results do not have statistical significance to correlate with a higher risk of thrombosis

Purpose. Even though there are no solid data regarding chemotherapy treated leukemia during pregnancy, the results based on short series reports show that the management of such condition can be safely achieved during the second and third trimester. We present three personal cases of pregnant women treated with cytostatic agents, two of them accidentally receiving complete chemotherapy during the entire pregnancy without malformative consequences. First case. A 19 yrs old woman diagnosed with chronic myeloid leukemia who conceived spontaneously and mistook the pregnancy signs for a relapse of the disease. During the pregnancy she continued the treatment, receiving until the fifth month an association of Hydroxyurea and alfa-interferon and afterwards switched to Imatinib until term. She presented at 38–39 weeks and delivered by cesarean section a little girl of 3510 g in a perfect state of health. The blood count of both mother and child were normal. Second case. A similar situation in a young woman with lymphoblastic acute leukemia under treatment with Vincristin, Methotrexat, Purinethol. She presented in advanced spontaneous labour at 33–34 weeks and delivered a little girl of 1700 g without malformative signs and normal blood count. Third case. A 17 years old girl who was diagnosed with acute myeloid leukemia at 29 weeks pregnancy. She received induction chemotherapy with Ara-C, due to the significant bone marrow infiltrate and disease induced disseminated intravascular coagulopathy. She presented premature uterine contractions at 32 weeks and delivered by cesarean section a premature boy of 1750g with Apgar score 8. The infant did not present any malformation (by clinical and ultrasound examination) and the blood count was normal. The studies have shown so far that in the case of chronic myeloid leukemia, the treatment with Imatinib was associated with 50% apparently normal live infants and that chemotherapy for acute leukemia during the second or third trimester may not require termination of pregnancy, because both remission and delivery of a normal infant are likely to be obtained.