23 results on '"Paro MM"'
Search Results
2. Fine needle aspiration cytology of hepatic and pancreastic tumours
- Author
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Kardum-Selin, Ika, Šušterčić, Dunja, Fabijanić, I, Jelić-Puškarić, B, Kušec, Rajko, Kardum-Paro, MM, Planinc-Peraica, Ana, Ostojić-Kolonić, Slobodana, Odak, D, Anić, P, Škegro, Dinko, Čolić-Cvrlje, Vesna, Papa, Branko, Jakšić, Branimir, Jonjić, Nives, and Kardum-Skelin, Ika
- Subjects
cytologic fine needle aspirates - Abstract
Imaging methods (ultrasound-US, computerised tomography-CT and magnetic resonance-MR) have ensured the visualisation of intraabdominal tumours (of liver, pancreas, kidneys, adrenal glands and retroperitoneal nodes) which are very rarely approachable on palpation, and consequently in guided aspiration. Aim of the study: 1. To analyse the adequacy of cytologic fine needle aspirates from intraabdominal organs and retroperitoneal nodes, 2. To estimate the occurrence rate of malignant lesions, 3. The possibility for typisation of epithelial and nonepithelial tumours by cytomorphology and additional technologies from a cytologic specimen. Patients and Methods: 1528 intraabdominal aspirates were examined: liver (696), pancreas (289). Fine needle aspiration was performed with the CHIBA needle. The preparations were analysed in standard stained smears by the May-Grunwald-Giemsa method, by determination of cellular markers by immunocytochemical analysis on smears and flow cytometry, cytochemical, cytogenetic and/or molecular analyses (PCR). Results: In 1480 patients fine-needle aspiration was performed with the US control, in 12 patients with CT, and in only 36 patients without image control. Insufficient material was obtained in 99 patients (6, 5%). Malignant cells were found in 648 cases in aspirates of both liver and pancreas. Of a total of 528 liver tumours, based on cytomorphology, completed with immunocytochemical markers, the epithelial, mesenchymal or embryonal cellular origin could be determined (92% epithelial, 6, 6% nonepitehelial origin and 1, 4% with suspicious features without precise differentiation of origin and/or type of tumours). In the tumors of epithelial group primary epithelial liver tumours numbered 21, 4% (101 hepatocellular and 3 cholangiocellular carcinoma), plus 306 metastatic ones, while in the epithelial tumors primary site was recognised in 15, 5% cases. In poorly differentiated tumors, the tumor aggressiveness was confirmed by high aneuploidy (DNA image cytometry) and by high proliferation status (AgNOR determination). In nonepithelial tumours the diagnosis of gastrointestinal stromal tumor (GIST) was made in 2, embryonal sarcoma in 1, angiosarcoma in 1, schwannoma in 1, leiomyosarcoma in 1, fibrosarcoma in 3, melanoma in 9 and 15 malignant lymphomas, while others could be diagnosed as sarcomas without precise differentiation. Out of 121 diagnosed malignant tumours of pancreas we found 112 carcinomas, 7 suspicious specimens and 1 mesenchymal tumour. Conclusion: Cytodiagnostic fine-needle aspiration of the liver and pancreas, completed with immunocytochemical markers on smears and flow cytometry, by cytogenetical and molecular analyses from the specimens obtained by cytologic aspiration, has been proved to be a reliable method in diagnostics and subtypisation of epithelial (primary as well as metastatic) and nonepithelial tumors. However, very often, some tumor markers reveal tissue co-expression and, as a rule, are not specific for only one organ. Despite this, their right combination improves the search for the primary site of the metastatic process or, potentially determine the additional diagnostic procedures (CT, US, MR).
- Published
- 2005
3. CLSI-based verification and de novo establishment of reference intervals for common biochemical assays in Croatian newborns.
- Author
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Friščić I, Perkov S, Radeljak A, Stipanović-Kastelić J, and Kardum Paro MM
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- Humans, Infant, Newborn, Reference Values, Croatia, Male, Female, C-Reactive Protein analysis, Creatinine blood, Aspartate Aminotransferases blood, Alanine Transaminase blood, Blood Chemical Analysis standards, gamma-Glutamyltransferase blood, Alkaline Phosphatase blood, Potassium blood, Magnesium blood, L-Lactate Dehydrogenase blood, Chlorides blood, Calcium blood, Blood Glucose analysis, Sodium blood, Bilirubin blood
- Abstract
Introduction: This study aimed to examine whether the Canadian Laboratory Initiative on Pediatric Reference Intervals (CALIPER) reference intervals for 19 commonly used biochemical assays (potassium, sodium, chloride, calcium, magnesium, inorganic phosphorous, glucose, urea, creatinine, direct and total bilirubin, C-reactive protein (CRP), total protein, albumin, aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP) and lactate dehydrogenase (LD)) could be applied to the newborn population of one Croatian clinical hospital., Materials and Methods: Reference interval verification was performed according to the CLSI EP28-A3c guidelines. Samples of healthy newborns were selected using the direct a posteriori sampling method and analyzed on the Beckman Coulter AU680 biochemical analyzer. If verification wasn't satisfactory, further procedure included de novo determination of own reference intervals by analyzing 120 samples of healthy newborns., Results: After the first set of measurements, 14/19 tested reference intervals were adopted for use: calcium, inorganic phosphorous, glucose, urea, creatinine, total bilirubin, CRP, total protein, albumin, AST, ALT, GGT, ALP and LD. A second set of samples was tested for 5 analytes: potassium, sodium, chloride, magnesium and direct bilirubin. The verification results of the additional samples for sodium and chloride were satisfactory, while the results for potassium, magnesium and direct bilirubin remained unsatisfactory and new reference intervals were determined., Conclusions: The CALIPER reference intervals can be implemented into routine laboratory and clinical practice for the tested newborn population for most of the analyzed assays, while own reference intervals for potassium, magnesium and direct bilirubin have been determined., Competing Interests: Potential conflict of interest None declared., (Copyright Croatian Society of Medical Biochemistry and Laboratory Medicine.)
- Published
- 2024
- Full Text
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4. Fragility spinal fractures among cirrhotic liver transplant candidates in Croatia.
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Jurina A, Delimar V, Giljević Z, Filipec Kanižaj T, Matković A, Vidović D, Jurjević N, Vidjak V, Duić Ž, Ćuk M, Japjec M, Dujmović T, Radeljak A, Kardum Paro MM, Vučić-Lovrenčić M, and Starešinić M
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- Adult, Humans, Absorptiometry, Photon methods, Bone Density, Bone Diseases, Metabolic, Croatia epidemiology, Cross-Sectional Studies, Lumbar Vertebrae diagnostic imaging, Renal Insufficiency, Chronic epidemiology, Liver Cirrhosis epidemiology, Liver Transplantation, Spinal Fractures diagnostic imaging, Spinal Fractures epidemiology
- Abstract
Introduction: Existing data on fragility spinal fractures prevalence in liver transplant candidates are scarce and inconsistent. This may be due to other comorbidities, besides hepatic osteodystrophy (HO), that contribute to bone loss and fragility fracture prevalence in chronic liver disease (CLD)., Objectives: The aim of this study was to investigate the prevalence of spinal thoracic and lumbar fragility fractures among cirrhotic, non-chronic kidney disease (CKD), non-diabetic liver transplant candidates and to explore their relationship with clinical characteristics, laboratory markers and dual-energy x-ray absorptiometry (DXA) results., Material and Methods: This cross-sectional observational study was conducted at Merkur University Hospital, Croatia, between February 2019 and May 2023. Adult patients with liver cirrhosis referred for liver transplantation were included. Patients with acute infection, CKD, diabetes mellitus, malignancies, inflammatory bone diseases and those on corticosteroid or antiresorptive therapy were excluded. Clinical, laboratory and radiological assessment was carried out and patients were accordingly allocated into non-fractured and fractured group for the purpose of statistical analysis., Results: A total of 90 patients were included in the study. There was 123 fractures, 87 (70.7 %) in the thoracic and 36 (29.3 %) in the lumbar region. Eighty-nine (72.4 %) fractures were grade 1, 31 (25.2 %) were grade 2 and 3 (2.4 %) were grade 3. Patients in the fractured group were significantly older (p < 0.001). No significant differences between fractured and non-fractured group according to laboratory and DXA parameters were noted. Subgroup with lumbar fractures had significantly lower bone mineral density values at L1-L4 region. Statistically significant negative correlation between bone specific alkaline phosphatase (BALP) and hip total BMD (rho = -0.414, p < 0.001) and spine total BMD (rho = -0.258, p = 0.014) values was found., Conclusion: Present study confirmed detrimental impact of CLD and HO on bone strength. DXA measurement correlated with the presence of lumbar fragility fractures. A combination of standard X-ray imaging and DXA is needed for adequate bone evaluation in pretransplant period and BALP could be useful for detecting HO in CLD. Searching for other risk factors and implementing bone turnover markers and additional imaging techniques for bone loss evaluation in liver transplant candidates is needed., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 Elsevier Ltd. All rights reserved.)
- Published
- 2024
- Full Text
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5. Diagnostic validation of two SARS-CoV-2 immunochromatographic tests in Slovenian and Croatian hospitals.
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Ifko M, Tkalčić Švabek Ž, Friščić I, Kardum Paro MM, Prkačin I, Đerek L, Livun A, and Skvarč M
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- Hospitals, Humans, Predictive Value of Tests, Sensitivity and Specificity, COVID-19, SARS-CoV-2
- Abstract
Aim: To diagnostically validate two point-of-care (POC) rapid antigen tests for SARS-CoV-2 by comparing their results with those of laboratory-based real-time polymerase chain reaction tests (RT-PCR)., Methods: The study enrolled 455 patients from two Slovenian and two Croatian hospitals. The NADAL COVID-19 Ag Test (Nal von Minden, Moers, Germany) and ALLTEST COVID-19 Antigen Test (Hangzhou ALLTEST Biotech Co., Ltd, Hangzhou, China) were diagnostically validated in emergency care departments of two Slovenian hospitals, while only ALLTEST COVID-19 Antigen Test was validated in two Croatian hospitals., Results: The antigen test results were in very good agreement with the RT-PCR results (Cohen's Kappa between 0.747 and 0.891 for the NADAL COVID-19 and between 0.820 and 0.954 for the ALLTEST COVID-19). The NADAL COVID-19 Ag Test had the sensitivity between 66.67% and 92.31%, with a negative predictive value between 85.51% and 99.2%. The ALLTEST COVID-19 Antigen Test had the sensitivity between 81.39% and 91.11%, with a negative predictive value between 85.45% and 98.78%., Conclusion: The antigen tests are practical and reliable screening assays for SARS CoV-2 in emergency care departments. Both antigen tests can be used as screening tests to reduce the number of patients waiting for RT-PCR results. Even more, they can be used to quickly isolate COVID-19 patients and reduce hospital transmissions.
- Published
- 2021
6. Laboratory professionals' attitudes towards ISO 15189:2012 accreditation: an anonymous survey of three Croatian accredited medical laboratories.
- Author
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Lapić I, Rogić D, Ivić M, Tomičević M, Kardum Paro MM, Đerek L, and Alpeza Viman I
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- Adult, Croatia, Female, Humans, Male, Quality Assurance, Health Care, Surveys and Questionnaires, Accreditation, Attitude of Health Personnel, Laboratories, Hospital standards
- Abstract
Introduction: Effective implementation and continual compliance with ISO 15189:2012 require ongoing commitment and active involvement of laboratory staff. Our aim was to assess attitudes regarding accreditation implementation by conducting a survey in three Croatian accredited medical laboratories., Materials and Methods: An anonymous survey consisting of 34 questions was distributed either electronically or in a paper form a week prior to scheduled annual audits. Distributions of answers regarding age, work experience, laboratory workplace, and education level and according to the respective laboratory were compared., Results: The overall response rate was 76% (225/297). Preference towards working in an accredited laboratory and a positive attitude were revealed by 70% and 56% participants, respectively, with better process documentation as the main advantage. Only 14% of responders considered themselves completely familiar with ISO 15189:2012. Total of 68% of responders felt that accreditation increases the usual workload, with excessive paperwork as the main contributor. Half of the responders declared partial agreement that accreditation requirements and expectations were clearly explained and claimed that their suggestions were taken into account only occasionally, which was especially emphasized by technical staff. The vast majority (89%) completely follow the prescribed protocols. Only 27% consider turnaround time monitoring useful. Competence assessment is considered efficient by 41% of responders. The majority (73%) prefer an online audit in times of COVID-19., Conclusions: Despite an overall positive attitude towards accreditation, further efforts are needed in providing better education about ISO 15189:2012 for technical staff and modifying formats of competence assessment, in order to achieve better adherence to ISO 15189:2012 requirements., Competing Interests: Potential conflict of interest None declared., (Croatian Society of Medical Biochemistry and Laboratory Medicine.)
- Published
- 2021
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7. Risk analysis of the preanalytical process based on quality indicators data.
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Flegar-Meštrić Z, Perkov S, Radeljak A, Kardum Paro MM, Prkačin I, and Devčić-Jeras A
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- Humans, Patient Safety, Risk Assessment, Risk Management, Clinical Laboratory Services standards, Diagnostic Errors prevention & control, Diagnostic Errors statistics & numerical data, Health Plan Implementation statistics & numerical data, Quality Indicators, Health Care statistics & numerical data
- Abstract
Background: Improving quality and patient safety in the medical biochemistry laboratory accredited according to the International Standard Organization (ISO 15189:2012) requires the patient-centered evaluation of errors based on the implementation of quality indicators (QIs) across the total testing process. Our main goal was to achieve quality improvement of the preanalytical process in an emergency laboratory which had the highest error rate using risk management principles., Methods: Failure mode and effects analysis (FMEA) was applied to analyze predefined preanalytical QIs and score laboratory failures for the failure demerit value (FDV), probability of failure (PF) and probability of failure remedy (PFR). Based on obtained scores (on a 10-point scale) risk priority numbers (RPNs) were calculated., Results: A total of five failure modes were identified in the preanalytic process. The calculated risks were "sample hemolysis" (RPN, 168),"misidentified samples" (RPN, 108),"samples clotted" (RPN, 90),"sample volume error" (RPN, 72) and "samples transported at inappropriate temperature" (RPN, 24). The activation of corrective risk-reducing measures for failure modes with RPN≥30 resulted in quality improvement with the significant decrease in reevaluated RPNs., Conclusions: The implementation of a preanalytical quality monitoring system based on observation of evidence-based QIs and patient-centered evaluation of errors through risk analysis with regular tailored education as well as implementing process improvements can effectively reduce preanalytical errors in the emergency laboratory and improve patient safety.
- Published
- 2017
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8. Fine-needle aspiration cytology yield as a basis for morphological, molecular, and cytogenetic diagnosis in alk-positive anaplastic large cell lymphoma with atypical clinical presentation.
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Bogdanic M, Ostojic Kolonic S, Kaic G, Kardum Paro MM, Lasan Trcic R, and Kardum-Skelin I
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- Anaplastic Lymphoma Kinase, Biomarkers, Tumor genetics, Biomarkers, Tumor metabolism, Biopsy, Fine-Needle, Diagnosis, Differential, Humans, Karyotype, Lymphoma, Large-Cell, Anaplastic genetics, Lymphoma, Large-Cell, Anaplastic metabolism, Male, Receptor Protein-Tyrosine Kinases genetics, Young Adult, Lymphoma, Large-Cell, Anaplastic pathology, Sarcoma pathology
- Abstract
ALK positive anaplastic large cell lymphoma is a T-cell lymphoma usually occurring in children and young adults. It frequently involves lymph nodes and extranodal sites and is associated with favorable prognosis. A 20-year old man was admitted for painful mass in the left axilla with overlying skin redness. Clinical presentation and US findings were highly suspicious for sarcoma. Definitive diagnosis was established cytolologically and using ancillary technologies from cytological samples. Fine needle aspiration cytology of tumor mass (lymph node conglomerate and surrounding tissue) show predominance of large, pleomorphic, atypical cells with large nuclei and vacuolised cytoplasm. Atypical cells immunocytochemically were positive for LCA, CD30, CD3, EMA, and ALK; negative for CD15 and CD56. NPM-ALK transcript was detected by reverse transcriptase-polymerase chain reaction (RT-PCT). Molecular analysis of TCRß and TCRγ genes demonstrated clonal TCR genes rearrangement. Complex karyotype with multiple numerical and structural changes was found on conventional cytogenetics. These findings excluded sarcoma and corroborated the diagnosis of ALK positive ALCL. Cutaneous involvement in ALCL can clinically mimic sarcoma, especially in cases with localized disease without B symptoms. In those cases, immunostaining, PCR, and conventional cytogenetics are helpful to exclude sarcoma. Diagn. Cytopathol. 2017;45:51-54. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)
- Published
- 2017
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9. Follicular and mantle cell lymphoma characteristics present simultaneously in the same lymph node.
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Korać P, Horvat T, Kardum Paro MM, Ajduković R, Džebro S, and Dominis M
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- Adult, B-Lymphocytes metabolism, Dendritic Cells metabolism, Humans, Immunohistochemistry, Immunophenotyping, Lymph Nodes metabolism, Lymphoma, Follicular complications, Lymphoma, Follicular diagnosis, Lymphoma, Follicular genetics, Lymphoma, Mantle-Cell complications, Lymphoma, Mantle-Cell diagnosis, Lymphoma, Mantle-Cell genetics, Male, B-Lymphocytes pathology, Biomarkers, Tumor genetics, Dendritic Cells pathology, Lymph Nodes pathology, Lymphoma, Follicular pathology, Lymphoma, Mantle-Cell pathology
- Abstract
Follicular lymphoma is composed of clonal germinal center B cells. It shows a follicular pattern lacking mantle zones, with a network of interfollicular dendritic cells. Transformation to more aggressive lymphomas is documented, but the only connections to mantle cell lymphoma are described cases of composite lymphoma consisting of these 2 entities. We discuss here a case of a lymph node harboring CD20, CD10, BCL2, BCL6, cyclin D1, CD5, Ki67, and SOX11 with CD21, showing an almost intact network of dendritic cells in one part of a lymph node, and CD20, CD5, SOX11, BCL6, cyclin D1, CD10, Ki67, and CD21 cells restricted to the mantle area in another part of the same lymph node. Both parts of the lymph node had BCL2 rearrangement, a lack of t(11:14)(q13;q32), the presence of SOX11 expression, and the same clonal band. The described case suggests heterogenous development of small cell lymphomas and indicates the possibility of differentiation regression.
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- 2013
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10. Neonatal hyperimmune T-cell reaction mimicking T-cell non-Hodgkin's lymphoma following BCG and hepatitis B co-vaccination.
- Author
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Dotlic S, Vranic S, Jakovljevic G, Ilic I, Kardum-Paro MM, and Dojcinov SD
- Subjects
- Adjuvants, Immunologic adverse effects, BCG Vaccine immunology, Diagnosis, Differential, Diagnostic Errors, Granuloma etiology, Granuloma pathology, Hepatitis B Vaccines immunology, Humans, Immune System Diseases etiology, Infant, Newborn, Lymphadenitis etiology, Lymphoma, T-Cell etiology, Male, Vaccines, Combined immunology, BCG Vaccine adverse effects, Hepatitis B Vaccines adverse effects, Immune System Diseases diagnosis, Lymphadenitis diagnosis, Lymphoma, T-Cell diagnosis, Vaccines, Combined adverse effects
- Abstract
We describe a case of a 2-week-old male infant who presented with a rapidly enlarging inguinal mass after having received both the bacille Calmette-Guérin (BCG) and hepatitis B vaccines at birth. The clinical picture raised suspicion of a neoplasm, and an excision biopsy was performed. It showed complete effacement of the lymph node architecture by a diffuse proliferation of monomorphic, mitotically active, and medium-sized T-cell blasts with strong expression of CD99. Coalescent necrotizing granulomas were also seen. The lymph node culture was negative for BCG. Upon expert review and additional molecular diagnostics, the initial pathological diagnosis of lymphoblastic T-cell lymphoma was changed to ectopic BCG lymphadenitis and hyperimmune post-vaccinal reaction. The atypical T-cell proliferation was most likely a result of the adjuvant effects of the co-administered vaccines. Post-vaccinal reactions usually involve the injection site or result in localized lymph node enlargements in the areas draining the inoculation site. This case highlights the importance of the clinical context for accurate interpretation of the pathological findings. In the setting of post-vaccinal lymphadenopathy, a biopsy is rarely needed but, when performed, should be interpreted with great caution.
- Published
- 2012
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11. [Significance of participation in programs of external quality assessment in molecular diagnostic--our experience].
- Author
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Paro MM, Siftar Z, Juretić D, and Flegar-Mestrić Z
- Subjects
- Croatia, Europe, Humans, Laboratories standards, Pathology, Molecular standards, Quality Assurance, Health Care
- Abstract
Harmonization of molecular diagnostic tests in laboratories in the Republic of Croatia has only just started. According to laboratory accreditation standard ISO 15189 participation in external quality assessment (EQA) schemes or programs is a prerequisite and support tool for clinical laboratory accreditation process. As there are no national quality assurance schemes yet, an European external quality assessment (EQA) scheme or program should be found. Because of variation in the molecular diagnostic test performance of clinical laboratories across Europe, EQA is recognized as a system whereby a set of reagents and techniques are assessed by an external provider making inter-laboratory performance comparability possible through already integrated recommendations and practice guidelines of molecular diagnostic test performance. Today, wide range of various EQA schemes and programs already in action have been available and most of them began within the last ten years. This paper is therefore intended to present and summarize the four-year EQA activities in the Institute of Clinical Chemistry, Merkur University Hospital, in three different international EQA schemes: United Kingdom National External Quality Assessment Scheme (UK NEQAS), the European Molecular Genetic Quality Network (EMQN) and Multi-National External Quality Assay program (EQUAL- qual)) and to point out their educational role in standardization of laboratory performance of any test intended for patient testing. from a laboratory point of view.
- Published
- 2011
12. [The unclassifiable myeloproliferative neoplasm--morphological, cytogenetic and clinical features].
- Author
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Borovecki A, Skrtić A, Paro MM, Lasan R, and Dominis M
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- Adult, Aged, Aged, 80 and over, Bone Marrow pathology, Cytogenetic Analysis, Female, Genes, abl genetics, Humans, Janus Kinase 2 genetics, Male, Middle Aged, Mutation, Myeloproliferative Disorders genetics, Myeloproliferative Disorders pathology, Myeloproliferative Disorders diagnosis
- Abstract
Myeloproliferative neoplasm, unclassifiable (MPN,U) has clinical, laboratory and morphological features of an MPN but fails to meet the criteria for any of the specific MPN entities. Because overlapping features, morphological findings in bone marrow, BCR-ABL1 fusion gene, V617F JAK2 mutation and cytogenetic abnormalities were analyzed in ten patients diagnosed with MPN,U. Bone marrow biopsy showed hypercellularity with trilineage myeloproliferation, dispersed megakaryocytes with mild pleomorphism and mature nuclei, and absence of reticulin fibrosis. All patients were BCL-ABL1 negative, while V617F JAK2 mutation was found in 6 of 8 patients. Trisomy 8 was found in two patients and t(6;12)(q12;p13) in one patient. Morphological features of MPN,U are nonspecific, however, in study cases they were most similar to diagnostic morphological features of polycythemiea vera. The high frequency of V617F JAK2 mutation in MPN,U cases analyzed revealed that its presence does not confirm a specific type of MPN.
- Published
- 2011
13. [Fine needle aspirate of lymph node as the analytical sample for immunophenotyping].
- Author
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Svencbir V, Anić V, Siftar Z, Paro MM, Kolonić SO, Bolanca IK, and Kardum-Skelin I
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- Clone Cells, Humans, Lymphoma, Non-Hodgkin immunology, Lymphoproliferative Disorders immunology, B-Lymphocytes immunology, Biopsy, Fine-Needle, Flow Cytometry, Immunophenotyping, Lymph Nodes pathology, Lymphoma, Non-Hodgkin diagnosis, Lymphoproliferative Disorders diagnosis
- Abstract
In modern clinical laboratory routine, cell analysis by flow cytometry means help in setting up the diagnosis by determination of B-lymphocyte clonality and thus separation of benign and malignant lymphoproliferative diseases. The aim of this study was to assess the value of cytologic diagnosis and adequacy of the material obtained by fine needle aspiration (FNA) of lymph nodes for flow cytometry analysis in cases of benign lesions and primary malignant lesions of lymph nodes. In addition, the aim was to determine B-lymphocyte clonality in different groups of benign and malignant lymph node lesions. The study was based on medical documentation, cytologic smears of FNA lymph node samples and results of flow cytometry immunophenotyping. A total of 239 patients were included over a one-year period. Patients were classified according to cytologic findings in the groups of non-Hodgkin's lymphoma of B cell origin (55%), benign lymphoproliferative disease (22%), undefined group of monomorphic population of lymphatic cells (16%), and the rest in the group of non-Hodgkin's non B cell origin. Study results showed FNA to be an appropriate method for obtaining sufficient numbers of cells for analysis by flow cytometry because there was no inadequate samples in our study group. In some cases of monomorphic lymphoid cell population, cytologic diagnosis was limited to small cell lymphomas, so determining the clonality by flow cytometry is crucial in separating malignant from benign lymphoproliferative disease. It is concluded that FNA associated with the flow cytometry method is a simple and safe method in the diagnosis of lymphoproliferative disease.
- Published
- 2011
14. [Flow cytometry and acute renal rejection confirmed by histopathologic analysis].
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Siftar Z, Sokolić I, Paro MM, Nazor A, Knotek M, Sabljar-Matovinović M, Vidas Z, and Flegar-Mestrić Z
- Subjects
- Acute Disease, Adolescent, Adult, Aged, Antilymphocyte Serum therapeutic use, Female, Graft Rejection pathology, Graft Rejection prevention & control, Humans, Immunosuppressive Agents therapeutic use, Kidney pathology, Lymphocyte Count, Male, Middle Aged, Young Adult, Flow Cytometry, Graft Rejection diagnosis, Kidney Transplantation
- Abstract
Transplantation of solid organs, tissues or hematopoietic cells is now standard in the treatment of patients with terminal stage disease in order to cure and improve the recipients' quality of life. The study included 54 patients having undergone single or multiple organ transplantation. All patients received a combination of immunosuppressant therapy consisting of corticosteroids, calcineurin inhibitor (cyclosporine; tacrolimus), anti-CD25 (daclizumab) and mycophenolate-mofetil. In 24 patients, acute rejection was stratified by histopathologic analysis of renal biopsy. Fifteen highly sensitized patients were administered antithymocyte globulin (ATG) therapy. Absolute count and percentage of B/T lymphocyte subsets, NK cells and CD25+ or CD69+ activated T cells were measured on a flow cytometer (EPICS XL, Coulter) using single platform standardized protocol. Upon ATG therapy, rapid decline to a very low level of T and NK cell lymphocyte count was observed, as well of B lymphocytes, resulting in redistribution of lymphocyte compartment. Between consecutive measurements, kinetic changes of lymphocyte subset numbers (absolute count or percentage) did not differ in a large spectrum of immune parameters between the groups with and without rejection episode and having received quadruple immunosuppressive induction and maintenance therapy. Immunologic monitoring must be initiated prior to transplantation and continued consistently and frequently post-transplantation. Such a program is expensive and time-consuming and stressful for the patient, therefore, prospective studies should identify whether treatment decisions can be based reliably on these immune parameters. Serial measurement of immune cell counts is necessary for maintenance of ATG therapy and could be useful for monitoring patient recovery.
- Published
- 2011
15. [Association of CD34 cell surface antigen expression with cytomorphological characteristics of acute promyelocytic leukemia blasts and clinical characteristics of patients: one center experience].
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Ostojić A, Pazur M, Siftar Z, Paro MM, Jelić-Puskarić B, Gredelj-Simec N, Radić-Kristo D, Kardum-Skelin I, Vrhovac R, and Jaksić B
- Subjects
- Adolescent, Adult, Aged, Antigens, Surface metabolism, Bone Marrow pathology, Female, Humans, Leukemia, Promyelocytic, Acute immunology, Leukemia, Promyelocytic, Acute pathology, Male, Middle Aged, Young Adult, Antigens, CD34 metabolism, Leukemia, Promyelocytic, Acute diagnosis
- Abstract
The aims of the study were to investigate the association between cytomorphology and immunophenotypic expression of CD34 cell surface antigen of blasts and their relationship with clinical and laboratory characteristics of patients with acute promyelocytic leukemia (APL). Sixteen consecutive patients (male 69% and female 31%) diagnosed with APL at Department of Hematology, Merkur University Hospital between August 1998 and December 2010 were included in the study. The mean age of patients was 43.9 (range: 18-78, SD 14.9). The patients' clinical and laboratory features, cytomorphological characteristics of APL-blasts and their immunophenotype determined by flow cytometry were analyzed. Patients were divided into two groups, CD34- and CD34+, and were then compared according to clinical and laboratory characteristics. There was no difference according to age, sex or white blood cell count between two groups. The mean value of hypogranular/agranular APL-blasts was markedly higher in CD34+ group than CD34- group (34%, range 9-60, SD 24.4 vs. 11.5%, range 0-38, SD 13.7), with borderline statistical significance (P=0.055). CD34- patients had significantly better overall survival than CD34+ ones (P=0.02). Patients without Auer rods detected in APL-blasts had higher CD34 expression (69.4% +/- 33.8) compared to patients with detected Auer rods (7.3% +/- 24.8), but statistical significance was not reached (p=0.053). Our results are consistent with the results of other published studies and point to the fact that higher CD34 expression and lower cytoplasmic granularity of APL-blasts are factors that seem to define a specific subgroup of APL patients. Together with other diagnostic tools currently available, they could be of value in planning treatment of APL patients.
- Published
- 2011
16. Flow cytometry immunophenotyping (FCI) of fine needle aspirates (FNAs) of lymph nodes.
- Author
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Paro MM, Siftar Z, Kardum-Skelin I, Sustercić D, Nazor A, Flegar-Mestrić Z, and Jaksić B
- Subjects
- Antigens, CD analysis, Antigens, CD immunology, Biopsy, Fine-Needle, Flow Cytometry methods, Flow Cytometry standards, Humans, Immunophenotyping standards, Lymphoma, Non-Hodgkin immunology, Retrospective Studies, Immunophenotyping methods, Lymph Nodes pathology, Lymphoma, B-Cell pathology, Lymphoma, Non-Hodgkin pathology
- Abstract
Flow cytometry immunophenotyping (FCI) has an important role in the clinic work-up of fine needle aspirates (FNAs) of lymph nodes. Its standardization has been defined by proposed analytical protocols and procedures used to assure proper analytical results also in those non-routine samples. In Institute of Clinical Chemistry, "Merkur" University Hospital, FCI is accredited method according to laboratory accreditation standard ISO 15189. According to this laboratory accreditation standard, participation in external quality assessment (EQA) programs is a prerequisite for assuring integrity and quality of the entire laboratory process. A critical analysis of our institutional experience in the feasibility of FCI of the material obtained by FNA of lymph nodes with suspected lymphoma represented the purpose of the study. During an eight-year period in Institute of Clinical Chemistry, "Merkur" University Hospital, a total of 1295 FNA analysis was done, 245 of them with a possible diagnosis of B-cell Non-Hodgkin lymphomas (B-NHL) formed the basis of the study. Lymphocytes were isolated on density gradient according to Boyum et al. The average feasibility of FNAs for FCI analysis was 86% (ranged 78-93%). An acceptable total cell number in FNAs for FCI analysis (4257) was established. In total population of respondents statistical significances in expressions of cellular antigens CD3, CD5, CD22, CD23, CD19 and CD5 on B-cells (CD5+CD19+) between patient's with final diagnosis of benign, reactive lymphoid proliferations and patient's with diagnosis of B-NHL were found. EQA results analysis showed that all results were either inside target values (X +/- 1SD) or inside accepted values (X +/- 2SD). Compatibility of the restriction of immunoglobulins light chains determinated by FCI and cytomorphology diagnosis depends on the choice of criterion values of the light chains ratio which determine the monoclonality. According to the matrix of shares of all classified data of retained neural network, ranges of diagnostic sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and prevalency of 82%, 72%, 93%, 48%, and 72% were produced. As a conclusion, FCI is a reliable methodology for phenotyping FNAs of lymph nodes with suspected B-NHLs detecting their clonality easily.
- Published
- 2010
17. Accreditation of medical laboratories in Croatia--experiences of the Institute of Clinical Chemistry, University Hospital "Merkur", Zagreb.
- Author
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Flegar-Mestrić Z, Nazor A, Perkov S, Surina B, Kardum-Paro MM, Siftar Z, Sikirica M, Sokolić I, Ozvald I, and Vidas Z
- Subjects
- Accreditation methods, Croatia, Documentation standards, Humans, Medical Audit, Quality Assurance, Health Care methods, Quality Control, Accreditation organization & administration, Chemistry, Clinical standards, Hospitals, University standards, Laboratories, Hospital standards, Quality Assurance, Health Care organization & administration
- Abstract
Since 2003 when the international norm for implementation of quality management in medical laboratories (EN ISO 15189, Medical laboratories--Particular requirements for quality and competence) was established and accepted, accreditation has become practical, generally accepted method of quality management and confirmation of technical competence of medical laboratories in the whole world. This norm has been translated into Croatian and accepted by the Croatian Institute for Norms as Croatian norm. Accreditation is carried out on voluntary basis by the Croatian Accreditation Agency that has up to now accredited two clinical medical biochemical laboratories in the Republic of Croatia. Advantages of accredited laboratory lie in its documented management system, constant improvement and training, reliability of test results, establishing users' trust in laboratory services, test results comparability and interlaboratory (international) test results acceptance by adopting the concept of metrological traceability in laboratory medicine.
- Published
- 2010
18. External quality assessment in clinical cell analysis by flow cytometry. Why is it so important?
- Author
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Siftar Z, Paro MM, Sokolić I, Nazor A, and Mestrić ZF
- Subjects
- Antigens, CD34 metabolism, Guidelines as Topic, HLA-DR Antigens metabolism, Humans, Lymphocyte Subsets metabolism, Lymphocyte Subsets pathology, Quality Control, Reproducibility of Results, Flow Cytometry standards, Immunophenotyping standards, Laboratories, Hospital standards, Leukemia pathology, Lymphoma pathology, Quality Assurance, Health Care
- Abstract
Participation in external quality assessment is an integral part of laboratory work and mandatory when the results have a clinical application, which is one of the requirements of standard 15189 for accreditation of medical laboratories. Institute of Clinical Chemistry, the first laboratory accredited for clinical cell analysis by flow cytometry in Croatia, participated in UKNEQAS for Leukocyte Immunophenotyping in 3 schemes: "Immune Monitoring", "CD34 Stem Cell Enumeration" and "Leukaemia Immunophenotyping". For sample processing on EPICS XL flow cytometer, lyse/no wash preparation technique with ammonium chloride (NH4Cl) or ImmunoPrep lysing reagent was employed. In "Immune monitoring" programme CD45/sideward light scatter (SSC) proposed gating strategy was adopted for lymphocyte subsets, while modified ISHAGE protocol was used for CD34+ cell enumeration. Absolute count determination was performed on flow cytometer using FlowCount beads solution. In the period from the beginning of 2006 until the middle of 2009 a total number of 100 stabilized whole blood samples were processed. The relative and absolute enumeration results for lymphocyte subsets were within tolerable limits, in 97.1 and 97.1% of cases, and 95 and 90% of CD34+ cell enumeration, respectively. In immune monitoring CD45/SSC proposed gating strategy is the most frequent analysis used (> 85% participants) and ISHAGE protocol for CD34+ cell determination with continuous rise from 76 to 83%. A number of participants who accept beads method for absolute count enumeration on flow cytometer get greater, 69 to 86%, while FlowCount was the second of bead-based techniques used (25 and 35%). Sample treatment in lyse/no wash technique using NH4Cl lysing solution was dominant procedure used by more than 1/3 participants, although its home made solution has replaced slowly by commercial reagents. The unacceptable results, 6 of 244, were obtained for 20 most frequently determined cell antigens in "Leukaemia Immunophenotyping" samples screened for leukaemia/lymphoma. Processing results of all participants showed that the deviation from laboratory guidelines and the use of older methods for cell identification, quantification of cell counting on haematology analyser, or usage an antibody conjugated with fluorochrome lesser fluorescence quantum often lead to an unacceptable result, although is noticeable trend to accept new referrals and protocols to reduce the inter-laboratory differences.
- Published
- 2010
19. FOXP1 and BCL2 show similar immunoenzymatic pattern in bone marrow trephines of chronic lymphocytic leukemia patients.
- Author
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Korać P, Vintar MG, Ajduković R, Kardum Paro MM, Jaksić B, and Dominis M
- Subjects
- Adult, Aged, Aged, 80 and over, B-Lymphocytes pathology, Bone Marrow pathology, Disease Progression, Female, Humans, Immunohistochemistry, Leukemia, Lymphocytic, Chronic, B-Cell pathology, Leukemia, Lymphocytic, Chronic, B-Cell physiopathology, Male, Middle Aged, Neoplasm Staging, Prognosis, B-Lymphocytes metabolism, Biomarkers, Tumor metabolism, Forkhead Transcription Factors metabolism, Leukemia, Lymphocytic, Chronic, B-Cell diagnosis, Proto-Oncogene Proteins c-bcl-2 metabolism, Repressor Proteins metabolism
- Abstract
Indolent B lymphoproliferative disorder, chronic lymphocytic leukemia (CLL) represents one of the most common hematologic diseases in the Western world. Although there are many disease development markers known so far, for example, B-cell lymphoma/leukemia (BCL) 2, new ones are needed for better understanding course of the disease. FOXP1 is known to be strongly expressed after B-cell activation. Its essential role in B-cell development suggested that it could also have a role in a various tumor B-cells. We have analyzed 74 bone marrow samples from B-CLL patients for presence of FOXP1 and its gene aberrations in tumor cells. Our results showed presence of FOXP1 protein mostly in the same tumor cells as BCL2 protein, and their specific immunostaining pattern. Diffuse immunostaining pattern of both proteins is present in patients with higher clinical stages of B-CLL and with some other markers that indicate worse outcome of the disease. Thus, FOXP1 and/or BCL2 immunostaining of bone marrow trephine sections could serve as an immunohistochemical marker in B-CLL.
- Published
- 2009
- Full Text
- View/download PDF
20. Immunohistochemical analysis of ZAP-70 expression in chronic lymphocytic leukemia.
- Author
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Korać P, Ajduković R, Kardum Paro MM, Jaksić B, and Dominis M
- Subjects
- Adult, Aged, Aged, 80 and over, Ataxia Telangiectasia Mutated Proteins, Biomarkers, Tumor biosynthesis, Cell Cycle Proteins genetics, Chi-Square Distribution, Chromosome Aberrations, DNA-Binding Proteins genetics, Female, Fluorescent Antibody Technique, Humans, Leukemia, Lymphocytic, Chronic, B-Cell diagnosis, Leukemia, Lymphocytic, Chronic, B-Cell genetics, Male, Microscopy, Fluorescence, Middle Aged, Mutation, Prognosis, Protein Serine-Threonine Kinases genetics, Reproducibility of Results, Sensitivity and Specificity, Tumor Suppressor Protein p53 genetics, Tumor Suppressor Proteins genetics, Immunohistochemistry methods, Leukemia, Lymphocytic, Chronic, B-Cell metabolism, ZAP-70 Protein-Tyrosine Kinase biosynthesis
- Abstract
This paper shows a protocol for the detection of ZAP-70 expression in B-CLL (B cell chronic lymphocytic leukemia) tumor cells by common immunohistochemical methods. The study was conducted on bone marrow trephine biopsies from 62 B-CLL patients at the time of diagnosis. Immunohistochemical reactions based on peroxidase and alkaline phosphatase reactions were used, as well as double immunofluorescent labeling for ZAP-70 detection as an indirect marker of mutated and unmutated CLL. Clinical relevance of the ZAP-70 expression detection method was assessed using chi2 test between ZAP-70 positivity data and other known prognostic factors, i.e., clinical and cytogenetics data. ZAP-70 was detected in 13 out of 62 patients. Statistically significant results were obtained for ZAP-70 positive cases and known indicators of worse prognosis. Immunohistochemical analysis supported by double immunofluorescent labeling, as shown here, is an easy and reliable technique for the detection of ZAP-70 expression in B-CLL tumor cells applicable in every hematopathology laboratory.
- Published
- 2009
- Full Text
- View/download PDF
21. [Single cell--definitive diagnosis! Where the profession ends and the art begins?].
- Author
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Kardum-Skelin I, Fabijanić I, Jelić-Puskarić B, Siftar Z, Kardum Paro MM, Trcić RL, Mahovlić V, Kusec R, and Seili-Bekafigo I
- Subjects
- Cell Biology, Cytological Techniques, Diagnosis, Computer-Assisted, Histocytochemistry, Humans, Molecular Diagnostic Techniques, Cytodiagnosis
- Abstract
Cell is a morphologically and functionally the tiniest living organism, present from the very beginning of life on the Earth. Specialized cell types make up specific tissues and organs of the human body. Cell itself and cell elements are liable to morphological, functional, phenotypic and genotypic alterations in various physiological and pathological states. These alterations are studied by cytodiagnosis to diagnose the disease at cellular level. Cytologic examinations belong to the group of morphological, non-aggressive or minimally invasive tests that are easy to perform for both the patient and the professional. In addition, these tests are highly reliable and preferred to the related diagnostic procedures for providing immediate orientation and definitive diagnosis, thus saving both time and money. With the introduction of adjunctive technologies such as cell-surface marker analysis, computer image analysis, molecular and cytogenetic technologies performed on cytologic smears, cytology has become an ever more important factor in the diagnosis, subtyping and prognosis of malignant tumors. Thorough knowledge of cell morphology is a basis for proper performance and understanding of cytology techniques. A cytologist needs to be familiar with clinical manifestations of the disease and to be informed on all relevant data on the patient and his current and previous medical history, in order to be able to issue findings that are understandable and usable to all clinicians and patients. It requires close collaboration between the cytology laboratory and the ward, and among the cytologist, the patient and the clinician. Good collaboration with other diagnostic professions such as pathology, laboratory, molecular and cytogenetic diagnosis is by no means less important. Cytology as a profession implies knowledge of the morphological characteristics of normal cells and cells in various physiologic states, along with due knowledge of the morphology, phenotypic and genetic features of pathologic alterations. However, synchronizing and combining cytologic morphology with other sophisticated diagnostic procedures to reach an accurate diagnosis, subtyping and prognosis of tumor disease is artistry indeed.
- Published
- 2008
22. Acute leukemia in patients with untreated chronic lymphocytic leukemia: a report of two cases with remarkably similar time cluster.
- Author
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Pavlisa G, Kolonic SO, Minigo H, Kardum-Skelin I, Kardum-Paro MM, and Jaksic B
- Subjects
- Acute Disease, Bone Marrow Examination, Chronology as Topic, Environmental Exposure adverse effects, Female, Humans, Immunophenotyping, Leukemia etiology, Leukemia, Lymphocytic, Chronic, B-Cell etiology, Leukemia, Radiation-Induced, Middle Aged, Uranium, Warfare, Leukemia diagnosis, Leukemia, Lymphocytic, Chronic, B-Cell diagnosis, Neoplasms, Multiple Primary diagnosis
- Published
- 2006
- Full Text
- View/download PDF
23. CD38 on B-cell chronic lymphocytic leukemia cells has higher expression in lymph nodes than in peripheral blood or bone marrow.
- Author
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Jaksic O, Paro MM, Kardum Skelin I, Kusec R, Pejsa V, and Jaksic B
- Subjects
- ADP-ribosyl Cyclase 1, Antibodies, Monoclonal metabolism, Cell Division, Flow Cytometry, Humans, Immunophenotyping, Lymph Nodes metabolism, Membrane Glycoproteins, Platelet Endothelial Cell Adhesion Molecule-1 biosynthesis, ADP-ribosyl Cyclase biosynthesis, Antigens, CD biosynthesis, Bone Marrow Cells metabolism, Leukemia, B-Cell metabolism, Leukemia, Lymphocytic, Chronic, B-Cell metabolism, Lymphatic Metastasis
- Published
- 2004
- Full Text
- View/download PDF
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