Evaluation of the JAK2V617F Mutational Burden in Patients with Philadelphia Chromosome Negative Myeloproliferative Neoplasms: A Single-center Experience.

The identification of the JAK2V617F mutation in several distinct myeloproliferative neoplasms (MPNs) raised the question how one single mutation incites expression of at least three different clinical phenotypes, i.e., polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF). In order to further evaluate already published data on the correlation between mutant JAK2V617F allele burden and specific hematological and clinical parameters, we tested the level of the JAK2 mutation in 134 JAK2+ patients with different MPNs. The patients were diagnosed according to the 2008 WHO criteria and followed for a median of 48 months. The JAK2 V617F quantification was done with a real time polymerase chain reaction (real time-PCR) method. The median allele burden was lowest in ET (25.8%), followed by 34.6% in PV and 51.8% in PMF patients (p<0.01). There was statistically significant association between the mutational load of 10.0-50.0% and blood count parameters in the PV patients (p<0.05). In PMF patients the mutational load was in correlation with older age and leukocyte count that were higher in patients with the mutational load of 10.0-50.0% and >50.0% compared to those with a mutational load of <10.0%. There were no statistically significant associations between the allele burden and blood counts in the ET cohort. Our study confirmed an association between the JAK2V617F allele burden and the distinct MPN phenotypes, indicating unfavorable prognosis in patients with a higher JAK2 allele burden. Our results suggest that JAK2 quantification should be incorporated in the diagnostic work-up of MPN patients as a useful tool for optimal treatment decision.

Haematology in the Republic of Macedonia: present situation and brief history.

The development of clinical haematology in Macedonia has taken place over the past nine decades. The greatest expansion of its development took place in the second half of the 20th century. The oficial start of clinical haematology dates from 1956, when the Department of Haematology was founded within the framework of the Internal Medicine Clinic in Skopje. In the beginning, haematology represented a form of virtual sub-specialty, but its expansion was so progressive and rapid that it reached the highest peaks of Yugoslav haematology in those times. The period from 1968 to 1979 was a period of integral development of haematology and blood-transfusion science in Macedonia. Nowadays, the autonomous Public Health Institution, the University Hematology Clinic, is a unique healthcare, educational and scientific establishment in the Republic of Macedonia in its field of work. The diagnostics algorithm comprises cyto-morphologic and cyto-chemical analysis, through immunologic characterization with the assistance of a flow cytometer, to sophisticated molecular analysis for detecting genetic abnormalities. The therapeutic approach is based upon modern poly-haemotherapeutic protocols, application of monoclonal antibodies, immuno-modulatory agents, molecular target therapy and the use of alogeneic and autologous transplantation of fresh bone-marrow and frozen haemopoietic stem-cells. The current motto of the Haematology Clinic is: always help those who seek help, provide precise and early diagnostics, and apply all up-to-date therapeutic strategies, scientific research, continual education and day-to-day implementation of the latest achievements in the field of haematology in daily practice.

Minimal screening analysis based algorithm for diagnosis and clinical stratification of patients with acute myeloid leukaemia (AML): single centre experience.

In this paper we present our results from a study designed in order to establish and standardize a diagnostic algorithm for acute myeloid leukaemia (AML) in the Republic of Macedonia. A total of 146 consecutive adult patients (>15 years) were enrolled in the study. First, we determined the correct lineage assignment of the blast cells and evaluated the incidence of the favourable PML/RARα, AML1/ETO, CBFß/MYH11 genetic markers among the AML cases. Additionally, the obtained results were correlated with patients' age, comorbidities, and performance status, and each single AML patient was stratified to effective treatment strategy. Our results showed that morphology and cytochemistry established a lineage in 132 (89.1%) of the patients, but not in 16 cases that presented as acute leukaemia, of which 7 were assigned as myeloid, and in two a non-haematopoietic malignancy was indicated with immunophenotyping. Mulitparameter flow cytometry immunophenotyping also changed the assigned lineage based on morphology and cytochemistry in 5 (3.3%) of the patients from lymphoid to myeloid and improved diagnosis in 21 (14.1%) cases. By using a reverse transcriptase-polymerase chain reaction (RT-PCR) essay 28 (23.1%) patients were classified in the prognostically favourable AML genetic group; 8 patients expressed the fusion transcript PML/RARα AML1/ETO and 15 CBFß/MYH11. Moreover, analyses of the age, performance status and comorbidities further strtified an additional 12.5% of the patients to a different risk-adapted therapy. The applied minimal screening-analysis-based diagnostic algorithm enabled improved and more precise diagnosis and clinical stratification in 37.2 % of AML patients from our study group.

A case report of aggressive adult neuroblastoma mimicking acute leukemia with fulminant course and fatal outcome.

A case of aggressive adult neuroblastoma mimicking acute leukemia with fulminant course and fatal outcome is described. Pancytopenia and circulating blasts cells at presentation suggested the diagnosis of acute leukemia in the previously healthy 38 years old Caucasian male patient, but flow-cytometry analysis of the bone marrow disclosed the correct diagnosis of neuroblastoma. The immunophenotype was CD45-/CD56+/CD9+ in around 50% of the mononuclear cells, indicating neuroectodermal origin of the malignant cells. Subsequently, the diagnosis was confirmed by immunohistochemical staining of a bone marrow biopsy. A review of the reported cases of neuroblastoma with leukemic features showed that several of them were misdiagnosed as having leukemia and that the diagnosis of neuroblastoma was made at autopsy examination, indicating that misdiagnosis may happen more often than is appreciated. It is in our opinion that the diagnosis of neuroblastoma should be considered in all cases of acute leukemia and pancytopenia, regardless of the age group of the patients.

Prognostic value of immunoglobulin variable heavy chain gene mutation status: long term follow-up in a series of chronic lymphocytic leukemia patients.

B-cell chronic lymphocytic leukemia (CLL) is a clinically heterogeneous disease, with many patients surviving for decades with minimal or no treatment, whereas others succumb rapidly to their disease despite therapy. Classical staging systems and laboratory features help predict survival in CLL, but they do not distinguish patients who will progress from those whose disease will remain indolent. In recent years, new molecular prognostic factors have emerged that have significantly improved prediction of the risk for disease progression. The mutational status of the immunoglobulin variable heavy chain genes (VH) is one of the major molecular prognostic factors. In this study we evaluated the association between the immunoglobulin VH gene mutation status and the clinical characteristics and outcome in 65 CLL patients that had been followed for a considerably long period at our institution. At diagnosis, patients with unmutated VH genes had higher median lymphocyte counts (P=0.001), higher total tumor mass score (P=0.001) and more often presented at an advance clinical stage (P=0.005) compared to patients utilizing mutated VH genes. Moreover, the median survival of patients with unmutated VH genes was considerably shorter (VH unmutated, 56 months, VH mutated, 125 months; P<0.001). These data confirmed the prognostic value of immunoglobulin VH genes mutational status in CLL, which divides the disease in two prognostic subsets in terms of overall survival and clinical characteristics of the disease. Analysis of the mutational status of the immunoglobulin VH genes may allow for an individualized approach to CLL treatment in the near future.