187 research outputs found
DETECTION OF CALR MUTATIONS USING HIGH RESOLUTION MELTING CURVE ANALYSIS (HRM-A); APPLICATION ON A LARGE COHORT OF GREEK ET AND MF PATIENTS
Background and Objectives
Somatic mutations in the calreticulin gene (CALR) are detected in approximately 70% of patients with essential thrombocythemia (ET) and primary or secondary myelofibosis (MF), lacking the JAK2and MPLmutations. To determine the prevalence of CALRframeshift mutations in a population of MPN patients of Greek origin, we developed a rapid low-budget PCR-based assay and screened samples from 5 tertiary Haematology units. This is a first of its kind report of the Greek patient population that also disclosed novel CALRmutants.
Methods
MPN patient samples were collected from different clinical units and screened for JAK2and MPLmutations after informed consent was obtained. Negative samples were analyzed for the presence of CALRmutations. To this end, we developed a modified post Real Time PCR High Resolution Melting Curve analysis (HRM-A) protocol. Samples were subsequently confirmed by Sanger sequencing.
Results
Using this protocol we screened 173 MPN, JAK2and MPLmutation negative, patients of Greek origin, of whom 117 (67.63%) displayed a CALRexon 9 mutation. More specifically, mutations were detected in 90 out of 130 (69.23%) essential thrombocythaemia cases (ET), in 18 out of 33 (54.55%) primary myelofibrosis patients (pMF) and in 9 out of 10 (90%) cases of myelofibrosis secondary to ET (post-ET sMF). False positive results were not detected. The limit of detection (LoD) of our protocol was 2%. Furthermore, our study reavealed 6 rare novel mutations which are to be added in the COSMIC database.
Conclusions
Overall, our method could rapidly and cost-effectively detect the mutation status in a representative cohort of Greek patients; the mutation make-up in our group was not different from what has been published for other national groups
Serum Soluble TACI, a BLyS Receptor, Is a Powerful Prognostic Marker of Outcome in Chronic Lymphocytic Leukemia
BLyS is involved in CLL biology and its low soluble serum levels related to a shorter time to first treatment (TFT). TACI is a BLyS receptor and can be shed from cells' surface and circulate in soluble form (sTACI). We investigated the impact of serum BLyS and sTACI levels at diagnosis in CLL patients and their relationship with disease parameters and patients' outcome. Serum BLyS was determined in 73 patients, while sTACI in 60. Frozen sera drawn at diagnosis were tested by ELISA. sTACI concentrations correlated with BLyS ( = −0.000021), b2-microglobulin ( = 0.005), anemia ( = −0.03), thrombocytopenia ( = 0.04), Binet stage ( = 0.02), and free light chains ratio ( = 0.0003). Soluble BLyS levels below median and sTACI values above median were related to shorter TFT ( = 0.0003 and 0.007). During a ten-year followup, sTACI levels, but not BLyS, correlated with survival ( = 0.048). In conclusion, we confirmed the prognostic significance of soluble BLyS levels with regard to TFT in CLL patients, and, more importantly, we showed for the first time that sTACI is a powerful prognostic marker, related to parameters of disease activity and staging and, more importantly, to TFT and OS
Antiretroviral activity of 5-azacytidine during treatment of a HTLV-1 positive myelodysplastic syndrome with autoimmune manifestations
Myelodysplastic syndromes (MDS) are often accompanied by autoimmune phenomena. The underlying mechanisms for these associations remain uncertain, although T cell activation seems to be important. Human T-lymphotropic virus (HTLV-1) has been detected in patients with myelodysplastic syndromes, mostly in regions of the world which are endemic for the virus, and where association of HTLV-1 with rheumatological manifestation is not rare. We present here the case of a 58 year old man who presented with cytopenias, leukocytoclastic vasculitis of the skin and glomerulopathy, and was diagnosed as MDS (refractory anemia with excess blasts - RAEB 1). The patient also tested positive for HTLV-1 by PCR. After 8 monthly cycles of 5-azacytidine he achieved a complete hematologic remission. Following treatment, a second PCR for HTLV-1 was carried out and found to be negative. This is the first report in the literature of a HTLV-1-positive MDS with severe autoimmune manifestations, which was treated with the hypomethylating factor 5-azacitidine, achieving cytogenetic remission with concomitant resolution of the autoimmune manifestations, as well as HTLV-1-PCR negativity. HTLV-1-PCR negativity may be due to either immune mediated clearance of the virus, or a potential antiretroviral effect of 5-azacytidine. 5-azacytidine is known for its antiretroviral effects, although there is no proof of its activity against HTLV-1 infection in vivo
Toxic iron species in lower-risk myelodysplastic syndrome patients:course of disease and effects on outcome
Atypical Chronic Myelogenous Leukemia, BCR-ABL1 Negative: Diagnostic Criteria and Treatment Approaches
Atypical chronic myelogenous leukemia (aCML), BCR/ABL1 negative is a
rare myelodysplastic/myeloproliferative neoplasm, usually manifested
with hyperleukocytosis without monocytosis or basophilia, organomegaly,
and marked dysgranulopoiesis. In this review, we will discuss the
classification and diagnostic criteria of aCML, as these have been
formulated during the past 30 years, with a focus on the recent advances
in the molecular characterization of the disease. Although this entity
does not have a definitive molecular profile, its molecular
characterization has contributed to a better understanding and more
accurate classification and diagnosis of aCML. At the same time, it has
facilitated the identification of adverse prognostic factors and the
stratification of patients according to their risk for leukemic
transformation. What is more, the molecular characterization of the
disease has expanded our therapeutic choices, thoroughly presented and
analyzed in this review article
Induction chemotherapy and post-remission imatinib therapy for de NovoBCR-ABL-positive AML
Factors affecting response to 5-azacytidine and prognosis of myelodysplastic syndrome. Is long-term survival a realistic goal?
The introduction of hypomethylating agents (HMAs) 5-azacytidine and decitabine has altered the prognosis of patients with myelodysplastic syndrome (MDS). Over the past few years, the International Prognostic Scoring System (IPSS) and the revised IPSS (IPSS-R) have been used both to define the prognosis of patients with MDS and to select patients to be treated with HMAs. Nevertheless, the prognosis of individual patients with MDS can differ considerably from the one calculated with the use of the above-mentioned prognostic systems. Thus, some patients may achieve long-term survival irrespective of their initial prognostic score. Several factors besides those used to define the IPSS/IPSS-R are analyzed in this review article; these include age and gender, the baseline hematologic characteristics, the comorbidities, the cytogenetic and molecular profile of the patients, as well as their response to treatment with 5-azacytidine. Thus, insight into a more personalized way of managing patients with MDS is given and long-term survival is set as a more realistic goal of treatment with 5-azacytidine. © 202
Significance of macrophage inflammatory protein-1 alpha (MIP-1 alpha) in multiple myeloma
Macrophage inflammatory protein-1 alpha (MIP-1 alpha) is a member of the
CC chemokine family and is primarily associated with cell adhesion and
migration. It is produced by myeloma (MM) cells and directly stimulates
osteoclast formation and differentiation in a dose dependent way. MIP-1
alpha protein levels were elevated in the bone marrow plasma of MM
patients and correlated with disease stage and activity. MIP-1 alpha was
also elevated in the serum of myeloma patients with severe bone disease
and correlated positively with bone resorption markers providing
evidence for a causal role of MIP-1 alpha in the development of lytic
bone lesions in MM. MIP-1 alpha has also been found to stimulate
proliferation, migration and survival of plasma cells. Mice, which were
inoculated with myeloma cells and treated with a monoclonal rat
anti-mouse MIP-1 alpha antibody, showed a reduction of both paraprotein
and lytic lesions. In addition, MIP-1 alpha enhanced adhesive
interactions between myeloma and marrow stromal cells, increasing the
expression of RANKL and IL-6, which further increased bone destruction
and tumor burden. Myeloma patients with high MIP-1 alpha serum levels
have poor prognosis. The positive correlation between MIP-1 alpha and
beta(2)-microglobulin that has been observed in MM patients at diagnosis
further supports the notion that MIP-1 alpha is not only a chemokine
with osteoclast activity function but is also implicated in myeloma
growth and survival. Therefore, MIP-1 alpha pathway may serve as a
target for the development of novel anti-myeloma therapies
Heparin induced thrombocytopenia: Contemporary therapeutic approaches in light of the new oral anticoagulants
Heparin induced thrombocytopenia (HIT) is a prothrombotic syndrome initiated by platelet- activating auto-antibodies with potentially devastating complications. Once the diagnosis of HIT is suspected, discontinuation of heparin and treatment with an alternative anticoagulant are mandatory. While established drugs for HIT are no longer available, parenteral factor Xa inhibitors, thrombin inhibitors and perhaps the direct oral anticoagulants provide additional treatment options. The aim of this review was to highlight the current clinical aspects regarding HIT focusing on the role of novel medications. © Schattauer 2015
Autoimmune hemolytic anemia during alpha-interferon treatment in a patient with chronic myelogenous lenkemia
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