Health-related quality of life in Croatian general population and multiple myeloma patients assessed by the EORTC QLQ-C30 and EORTC QLQ-MY20 questionnaires

Background. The impact of disease and treatment on the patient's overall well-being and functioning is a topic of growing interest in clinical research and practice. The aim of this study is to obtain reference data on quality of life of Croatian general population. Further, we aim to assess the impact of the disease and its primary systemic treatment on their health related quality of life (HrQoL) in multiple myeloma (MM) patients. Patients and methods. Participants for the first part of the study were randomly selected from adult Croatian population. In the clinical part of the study MM patients were included as prospectively diagnosed within two years in two major Croatian haematological centres. The EORTC QLQ-C30 in both trials and QLQ-MY20 in MM patients only were applied for HrQoL assessment. Results. Gender, age and place of residence have great impact on quality of life scores in Croatian population. The MM patients at the time of diagnosis have lower QLQ-C30 scores for global quality of life, functional and symptom scale scores, as well as single items. The type of disease followed by the choice of therapy options are important HrQoL determinants. Conclusions. The norm values available now for Croatian population will help to interpret HrQoL for clinicians and aid in planning cancer care interventions. This study identified treatment effect consistent with those from other observational studies and provided new data on HrQoL across two different treatment choices for MM patients

Detection of t(14;18) by PCR of IgH/BCL2 Fusion Gene in Follicular Lymphoma from Archived Cytological Smears

According to WHO classification follicular lymphoma (FL) is a neoplasm composed of follicle centre (germinal centre) B-cells, which usually has at least a partially follicular pattern. Bone marrow (BM) infiltration by lymphoma occurs in 40-70% of cases at the time of diagnosis. The characteristic chromosomal translocation of follicular lymphoma is t(14;18)(q32;q21) with transposition of BCL2 oncogene to the regulatory region of immunoglobulin heavy chain gene IgH. Aim of this study was to determine the frequency of PCR detection of IgH/BCL2 in DNA samples isolated from archival cytological slides of lymph node aspirates, bone marrow and/or peripheral blood (PB) obtained from patients with histologicaly confirmed follicular lymphoma using primers and protocol proposed by BIOMED-2 consortium. We also compared molecular with cytomorphological findings in bone marrow/peripheral blood and tested this method of detection of IgH/BCL2 molecular marker in monitoring minimal residual disease (MRD) in routine clinical setting. DNA was successfully isolated from all archival cytological slides obtained by fine needle aspiration of lymph nodes as well as from 75% of smears of bone marrow aspirates from 19 patients. Fusion oncogene was detected in 10 of 19 patients (52%). For patients with PCR IgH/BCL2 positive lymph nodes, molecular test found BM infiltration in 5 cases (83%), while cytomorphology detected infiltration in three of eight cases (37%) available for comparison. May-Grünwald-Giemsa stained cytological smears can be used for PCR-based ancillary methods and the rate of detection of IgH/BCL2 rearrangement is similar to results reported for paraffin-embedded tissues. For patients with detectable baseline molecular marker, PCR is a highly suitable method for detection of bone marrow involvement and monitoring MRD

Comparison of sensitivity of nested PCR and quantitative PCR in Bcr-Abl p210 transcript detection in chronic myelogenous leukemia

Uvod: Za otkrivanje minimalne ostatne bolesti u bolesnika s kroničnom mijeloičnom leukemijom (KML) koji su postigli potpunu kliničku remisiju i potpun citogenetski odgovor može se primijeniti ugnježđena PCR (engl. nestedPCR, nPCR) i kvantitativna PCR u stvarnom vremenu (engl. quantitative real-time PCR, qPCR). Cilj liječenja je postizanje molekularne remisije, pa postizanje visoke osjetljivosti molekularne pretrage ima presudnu ulogu i kliničku primjenu. Usporedili smo razinu osjetljivosti nPCR i qPCR u otkrivanju BCR-ABL p210 prijepisa u modelu razrjeđenja Bcr/Abl-pozitivnih stanica. Materijal i metode: Za određivanje razine osjetljivosti načinjena su serijska razrjeđenja stanične linije K562 (Bcr-Abl pozitivne) sa staničnom linijom NB4 (Bcr-Abl negativnom) (raspon razrjeđenja pozitivnih stanica: 10-3-10-7). Izolirani uzorci RNA prepisani su u cDNA i testirani na p210 prijepis pomoću nPCR i qPCR. Objema metodama također su ispitani uzorci koštane srži i periferne krvi dvoje bolesnika s KML na terapiji imatinib-mesilatom. Rezultati: U testu staničnog razrjeđenja nPCR je pokazala osjetljivost za otkrivanje Bcr-Abl pozitivnih stanica od 10-5, dok je qPCR pokazala osjetljivost od 10-6. Obje su metode otkrile p210 prijepise u uzorcima koštane srži bolesnika s KML. Međutim, qPCR je uz to otkrila prijepise i u uzorcima periferne krvi, no uz nižu razinu prijepisa u usporedbi s uzorcima koštane srži. Zaključak: Iako se često navodi kako je nPCR za otprilike 1 log osjetljivija od qPCR, u našem pokusu s razrjeđenjem na biljeg pozitivne stanične linije K562 dokumentirali smo višu osjetljivost za standardiziranu metodu qPCR, koja je razvijena za potrebe Europskog programa za borbu protiv karcinoma.Background: For minimal residual disease detection in chronic myelogenous leukemia (CML) patients who have achieved complete clinical remission and complete cytogenetic response, nested PCR (nPCR) and quantitative realtime PCR (qPCR) can be used. Achieving of molecular remission is the goal of therapy, so it is of critical importance and clinical utility to obtain high sensitivity of molecular testing. We compared the level of sensitivity of nPCR and qPCR in the detection of BCR-ABL p210 transcripts in a Bcr/Abl-positive cell dilution model. Materials and Methods: For determination of sensitivity level, serial dilutions of K562 cell line (Bcr-Abl-positive) in NB4cell line (Bcr-Abl-negative) were made (range of dilution of positive cells: 10-3-10-7). Isolated RNA samples were transcribed into cDNA and tested for p210 transcript by nPCR and qPCR. Bone marrow and peripheral blood samples of two CML patients on imatinib mesylate therapy were also tested by both methods. Results: In the cell dilution test, nPCR showed sensitivity for detecting Bcr-Abl positive cell of 10-5, and qPCR showed a sensitivity of 10-6. Both methods detected p210 transcripts in bone marrow samples of CML patients. However, qPCR also detected transcripts in peripheral blood samples, with a lower transcript level in comparison to bone marrow samples. Conclusion: Although frequently quoted as nPCR being by approximately 1 log more sensitive than qPCR, in our marker-positive cell line K562 dilution experiment we documented higher sensitivity of the standardized Europe Against Cancer Program developed qPCR method

The Accuracy of Fine Needle Aspiration Cytology and Flow Cytometry in Evaluation of Nodal and Extranodal Sites in Patients with Suspicion of Lymphoma

Today lymphomas are defined according to a combination of morphology, immunophenotype, genetic features and clinical presentation, so beside the pure cytomorphologic analysis in diagnosis of lymphoma ancillary techniques such as cytochemistry, immunocytochemistry, molecular diagnosis and flow cytometry (FC) are often used. Our goal was to determinate how is information given by fine-needle aspiration cytology (FNAC) and FC correlated with pathohistologic diagnosis and to evaluate ability to diagnose and subclassify malignant lymphomas by FNAC and FC. This study is a retrospective chart review of patients with suspicion of lymphoma processed at University Hospital Dubrava in Zagreb. After analysis 50 patients fulfilled inclusion criteria for this study (FNAC diagnosis with or without FC and consecutive confirmatory pathohistological diagnosis). When analyzing accuracy of FNAC according to suspicion of lymphoma or NHL and differential diagnosis lymphoma sensitivity was 97.7%, specificity 85.7% and the diagnostic accuracy was 96%. When analyzing accuracy of FNAC according to the subclassification of lymphoma, sensitivity was 74.4%, specificity 85.7% and the diagnostic accuracy 76%. Combined FNAC and FC improved sensitivity, positive predictive value, negative predictive value and diagnostic accuracy. Sensitivity was 79.1% and the diagnostic accuracy 80%. We have shown that these methods can distinguish benign lymphadenopaties from lymphomas and also subclassify lymphomas and quickly provide clinicians with that information

The degree of anisocytosis predicts survival in patients with primary myelofibrosis

Introduction: Red cell distribution width (RDW) provides a quantitative measure of anisocytosis and it is associated with the presence of subclinical systemic inflammation and a poor outcome in a variety of diseases when elevated. Anisocytosis is a feature of primary myelofibrosis (PMF) but it’s prognostic role in PMF has not yet been evaluated. ----- Patients and methods: 33 newly-diagnosed patients with PMF were analyzed. Baseline RDW values were obtained in addition to CRP, LDH, complete blood count, iron metabolism parameters and JAK2 V617F mutational status. Patients were staged according to IPSS prognostic scoring system, liver and spleen size were assessed by palpation. ----- Results: Median RDW was 19.0% (15.2%-22.5%). RDW correlated significantly with hemoglobin level (p=0.005), CRP (p=0.031), spleen size (p=0.036) and IPSS score (p=0.003). Patients with more pronounced anisocytosis had an inferior overall survival (OS) – very-high RDW (≥19.0%) vs. high RDW (15.1%-18.9%) subgroup, HR 5.37, p=0.002. RDW remained significantly associated with OS (p=0.002) in a multivariate model including IPSS score, hemoglobin level and CRP. ----- Conclusion: A higher degree of anisocytosis is associated with more advanced disease features and a decreased overall survival. RDW encompasses standard prognostic score and may help in the rapid detection of patients with an unfavorable prognosis