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Mircea Cucuianu, Ioana Brudasca, Luminita Plesca, Gyorgy Bodizs, Dan Colhon and Andrei Cucuianu

Abstract

Protein C is a vitamin K-dependent serine protease secreted by the hepatocytes as an inactive zymogen and activated by thrombin bound to endothelial thrombomodulin. An endothelial protein C receptor (EPCR) is involved in both activation and enhancement of protein C activity, resulting in proteolytic degradation of clotting factors Va and VIIIa, thereby providing an efficient anticoagulant mechanism. Evidence was also provided that proinflammatory cytokines would impair the endothelia-mediated activation and activity of the Protein C system by inducing an internalization and proteolytic degradation of thrombomodulin and by shedding EPCR from the surface of endothelial cells membrane. Clinical and experimental studies also emphasized that an inflammatory acute phase reaction is accompanied by a commuted hepatic protein synthesis leading to an increase of plasma fibrinogen, factor VIII:C and of α1 protease inhibitor, while the plasma level of protein C zymogen decrease. On the other hand infusions of activated protein C were reported to protect from a toxico-septic shock by exerting not only anticoagulant but also anti-inflammatory effects.

Open access

Elena-Cristina Selicean, Mariana Patiu, Andrei Cucuianu, Delia Dima and Minodora Dobreanu

Abstract

Morphological and immuno- flow cytometry assisted analysis of peripheral blood and bone marrow are mandatory investigations in the diagnosis of acute leukemia. Cytology and immunophenotyping complement each other primarily because they have as common object malignant cell phenotype as a whole. The aim of our study was to analyze correlations between cytology and immunophenotyping on a group of patients investigated for acute myeloid leukemia. In our study the degree of correlation between blast percentage determined by cytology and immunophenotyping was low (r=0.049). The degree of correlation between myeloperoxidase positivity in cytochemistry and immunophenotyping was also low, with better results for cytochemistry. Expression of immunophenotypic markers was consistent with the composition of our group regarding French-American-British classes, except for HLA-DR (49.0%), TdT (3.77%), CD14 (5.66%), CD15 (5.66%). We also discuss the importance of interpreting with caution positivity for erythroid and megakaryocytic markers and differential diagnosis of cases simultaneously expressing CD7 and CD56. In conclusion, interpretation of immunophenotyping by flow citometry, done in close conjunction with morphology, is mandatory to facilitate the use of optimized sample processing methods and of standardized panels, for both appropriate diagnosis and follow-up.

Open access

Bogdan Fetica, Ana-Maria Fit, Luminița Blaga, Annamaria Fulop, Bogdan Pop, Delia Dima, Andrei Cucuianu and Ljubomir Petrov

Abstract

Chronic lymphocytic leukemia (CLL) has a heterogeneous clinical course. Among useful markers in identifiyng patients with poor outcome are unmutated IgVH, ZAP-70 and CD38 expression. Both ZAP-70 and CD38 were shown to be capable of identifying aggressive CLL.

We analysed data from 35 patients diagnosed with CLL based on morphological and immunophenotypical criteria. In all cases peripheral blood immunophenotyping was performed as initial diagnostic test. Immunohistochemical expression of ZAP-70 and CD38 was evaluated on 21 cases of lymph node biopsies and 14 cases of bone marrow biopsies, performed at the time of diagnosis. In addition in-situ hybridization for EBER-1 was evaluated.

The median age of patients was 60 years and we noted a slight male predominance. The immunophenotypic criteria (C23+, CD5+, CD20+, CD10-, CD3-, cyclinD1-) for B-cell CLL were achieved in all 35 patients. We found that CLL cases showing expression of both markers (ZAP-70+CD38+ patients) are characterised by an unfavourable clinical course as compared with cases that did not show expression of markers (ZAP-70-CD38- patients). Our data showed significant differences in terms of overall survival at 5 years between the two groups. We also found statistically significant differences between patients ZAP-70-CD38- and patients with one or both positive markers (ZAP-70+ and/or CD38+).

Prognostic information given by ZAP-70 and CD38 could be used in guiding treatment decisions and they probably should be recommended to all patients with B-CLL in trying to obtain a more clear profile of the disease at the time of diagnosis.

Open access

Delia Dima, Adrian P. Trifa, Mariana Paţiu, Cristian S. Vesa, Ioana C. Frinc, Ljubomir Petrov and Andrei Cucuianu

Abstract

Introduction. Since the introduction of the tyrosine kinase inhibitor (TKI) imatinib mesylate (IM) in the treatment of chronic myeloid leukemia (CML), a dramatic improvement in hematologic, cytogenetic and molecular responses was noted. Also, the overall survival increased significantly. Unfortunately, in certain patients, resistance to TKI develops relatively early, especially due to point mutations in the ABL kinase domain, among which the T315I mutation confers resistance to all three currently available TKIs (imatinib, dasatinib, nilotinib). Methods. We performed a prospective study on 74 patients diagnosed with chronic phase CML, for whom we analyzed the T315I mutation. Mutational analysis was performed using ARMS-PCR (with subsequent confirmation by direct sequencing) at regular intervals of 6 months or in case of suboptimal response, loss of response or progression. Correlations between the T315I mutation and disease characteristics, response to treatment and survival were analyzed. A comparative analysis between patients positive and negative for the mutation was performed. The patients were followed and evaluated according to European Leukemia Net (ELN) criteria. Results. T315I mutation was detected in 3 patients (4.05%) and its presence was correlated with younger age at diagnosis, second line TKI therapy, progressive disease and decreased survival from the moment of detection. Conclusions. ARMS-PCR is a sensitive, easy to use method for the detection of T315I mutation in chronic phase CML patients