Citomorfološka i imunocitokemijska analiza u dijagnostici T i B nodalnih i Hodgkinovih limfoma [Cytomorphologic and immunocytochemical analyses in nodal T- and B-cell and Hodgkin´s lymphoma diagnostics]

Introduction: According to the World Health Organization classification of lymphoid neoplasms, it is important to recognize cell morphology and reveal its phenotype, then combine it with different genotypic information and clinical data to provide appropriate therapy. In theory, this enables cytomorphology, not just pathology, to serve as a primary morphologic method in evaluation of lymphoma patients. ----- Aim: To assess the efficacy and reproducibility of FNA and immunocytochemistry based lymphoma diagnostics, estimate usefulness in diagnosing T-, B-cell or Hodgkin´s lymphoma, and classify it upon the WHO classification. ----- Patients and methods: Specimens of 250 biopsy proven lymphoma patients, newly recognized or with relapsing disease, in whom FNA preceded other procedures were included in the study, regardless of it´s adequacy. Smears were stained with May - Grőnwald Giemsa, and immunocytochemical analyses were performed; those diagnosed as lymphoma were classified according to WHO classification. Two blindly performed consecutive cytomorphological analyses, indicated as cytomorphologist A and B, were compared with histopathology. Data obtained were submitted to ROC analysis, χ2 or Fischer exact test along with descriptive statistical methods. ----- Results: Eleven samples were found inadequate by each cytomorphologist, in six patients histopathologically diagnosed as cHL, in five as DLBCL and in one as CLL/SLL. Compared with histopathologic diagnoses exact agreement in lymphoma subtype was reached in 197/239 (82.4%) patients. Cytomorphologist A diagnosed B-cell lymphomas with 99.2% sensitivity and 99.0% specificity; T-cell lymphomas with 95.8% sensitivity and 98.4% specificity; 80 Hodgkin´s lymphoma with 95.0% sensitivity and 98.8% specificity. Cytomorphologist B diagnosed B-cell lymphomas with 100% sensitivity and 99.0% specificity; T-cell lymphomas with 100% sensitivity and 98.9% specificity; Hodgkin´s lymphoma with 95.0% sensitivity and 100% specificity. Statistically significant difference (p≤0.05) was found comparing histopathologic and cytomorphologic diagnoses in some subtypes of B and T-cell lymphomas (NMZL, FL, ALCL), but not comparing diagnoses between two cytomorphologists. ROC analysis (confidence interval 95%) proved high reliability of the method. ----- Conclusion: FNA, corroborated with immunocytochemistry, is an accurate method in the diagnosing T-, B-cell and Hodgkin`s lymphoma, as well as in lymphoma subtypes with characteristic cytomorphologic picture, such as CLL/SLL, BL, precursor T-cell lymphoma, PTCL and MF progressing to lymph node. High rate of consensus diagnoses between two cytomorphologists indicates reproducibility of the method

FNA based diagnosis of head and neck nodal lymphoma [Citomorfološka dijagnoza limfoma u području glave i vrata]

Fine-needle aspiration (FNA) biopsy has become a well established technique in the diagnosis, staging, and follow-up of patients with head and neck lesions. As in lymphoma diagnostics, FNA serves as a screening method in evaluating potentially affected lymph node for open or core biopsy. According to the World Health Organization classification of lymphoid neoplasms, today it is important to recognize cell morphology and reveal its phenotype, then combine it with different genotypic information and clinical data to provide appropriate therapy. The aim of this study was to assess the efficacy of FNA and immunocytochemistry based lymphoma diagnostic in head and neck region. We conducted a retrospective study during a period of three years where cases with either FNA diagnosis or clinical suspicion of newly recognized or relapsing lymphoma were reviewed. In the study were included patients that were referred to our laboratory from hematology department, in whom head and neck lymphadenopathia was found and lymph node FNA preceded other procedures. Two hundred eighty-five aspirations from 248 patients fulfilled study criteria. Adequate specimens were diagnosed as lymphoma in 100 cases (36%), in 65 male and 35 female patients, 76 in patients with newly discovered disease and 24 in patients with prior lymphoma diagnosis. Overall sensitivity of FNA specimens in the diagnosis of head and neck lymphomas was 90%, specificity 88%, predictive value of a positive result 97%, and predictive value of negative result 61%. Based on our results FNA corroborated with immunophenotyping by immunocytochemistry can be method of choice in primary lymphoma diagnosis as a method complementary to histopathology in lymphoma diagnostics

STANDARDISED FORM TO REPORT THE FINDING OF CYTOLOGICAL URINALYSIS – GUIDELINES OF THE CROATIAN SOCIETY FOR CLINICAL CYTOLOGY

Radi jasnijeg priopćavanja rezultata citološke analize urina i izjednačavanja terminologije članovi Hrvatskog društva za kliničku citologiju izradili su standardizirani obrazac za priopćavanje nalaza citološke analize urina. Prihvaćene su dijagnostičke kategorije Pariške klasifi kacije citologije urina te dodani opisi netumorske problematike urološke citologije. Predviđeno je da nalaz sadržava podatke o pacijentu, vrsti analiziranog uzorka, makroskopski pregled, metode obrade uzorka, primjerenost uzorka, citomorfološki opis i semikvantitativnu procjenu epitelnih i upalnih stanica, mikroorganizme, nestanične elemente, prisutnost i morfologiju eritrocita, dijagnozu prema Pariškoj klasifikaciji, komentar i zaključak te preporuke.In order to report clearly the results of cytological urinalysis and to standardise the terminology, the members of the Croatian Society for Clinical Cytology have elaborated a standardised form to report the finding of cytological urinalysis. The diagnostic criteria of the Paris Classification of Urinary Cytology have been accepted and further nonmalignant challenges have been added. The report contains information on the patient, type of specimen, macroscopic assessment, method of processing, adequacy of specimen, cytomorphological description and semi-quantitative analysis of epithelial and inflammatory cells, non-cellular elements, existence and morphology of erythrocytes, diagnosis in line with the Paris classification, comment, conclusion and recommendation

Apoptosis of Leukemic Cells: A Case Report

Transformation of leukemic cells is associated with delay in maturation and in apoptosis, and to altered responsiveness to growth factors. However, some studies have revealed that Fas (CD95/APO1) which mediates apoptotic signal and decrease of anti-apoptotic Bcl-2 are frequently observed in acute myeloid leukemia (AML) M4/M5 leukemic cells. The aim of the study was to compare cytomorphology and cytochemistry of bone marrow (BM) apoptotic leukemic cells to preserved peripheral blood (PB) leukemic cells in our patient, a 76-year-old man with AML-M5b treated at Zagreb University Hospital Center. BM and PB of the AL patient were analyzed after Pappenheim and cytochemical stainings, and leukemic cells were classified according to FAB and WHO classification. Analysis of PB revealed leukocytosis and 80–90% monocytic cells (46% monoblasts, 29% promonocytes and 11% monocytes). Only a few preserved monoblasts and promonocytes were found in BM, together with numerous morphologically altered cells with characteristic chromatin condensation and pyknosis of nucleus, as well as nuclear fragmentation and formation of apoptotic bodies. Thus, cytomorphology of PB leukemic cells pointed to proliferation of immature monocytic cells, and cytomorphology of BM to cell apoptosis. Cytochemistry of PB monocytic cells and BM apoptotic cells confirmed monocytic cell lineage because esterase was strongly positive in almost all BM apoptotic leukemic cells and PB leukemic cells, and esterase was completely inhibited with sodium fluoride. On the basis of these findings, AML-M5b was diagnosed in our patient. There are many possible explanations for our observation of BM leukemic cell apoptosis in a patient with AML-M5. The most reliable one is that apoptosis was induced ex vivo after BM aspiration in course of the air drying of BM specimen before staining. Mass BM leukemic cell apoptosis that was recorded in contrast to numerous preserved leukemic cells in PK could be probably connected to unfavorable ratio of relatively low concentration of cytokines in relation to high leukemic cell number in BM aspirated cytologic specimen

FNA Based Diagnosis of Head and Neck Nodal Lymphoma

Fine-needle aspiration (FNA) biopsy has become a well established technique in the diagnosis, staging, and follow-up of patients with head and neck lesions. As in lymphoma diagnostics, FNA serves as a screening method in evaluating potentially affected lymph node for open or core biopsy. According to the World Health Organization classification of lymphoid neoplasms, today it is important to recognize cell morphology and reveal its phenotype, then combine it with different genotypic information and clinical data to provide appropriate therapy. The aim of this study was to assess the efficacy of FNA and immunocytochemistry based lymphoma diagnostic in head and neck region. We conducted a retrospective study during a period of three years where cases with either FNA diagnosis or clinical suspicion of newly recognized or relapsing lymphoma were reviewed. In the study were included patients that were referred to our laboratory from hematology department, in whom head and neck lymphadenopathia was found and lymph node FNA preceded other procedures. Two hundred eighty-five aspirations from 248 patients fulfilled study criteria. Adequate specimens were diagnosed as lymphoma in 100 cases (36%), in 65 male and 35 female patients, 76 in patients with newly discovered disease and 24 in patients with prior lymphoma diagnosis. Overall sensitivity of FNA specimens in the diagnosis of head and neck lymphomas was 90%, specificity 88%, predictive value of a positive result 97%, and predictive value of negative result 61%. Based on our results FNA corroborated with immunophenotyping by immunocytochemistry can be method of choice in primary lymphoma diagnosis as a method complementary to histopathology in lymphoma diagnostics

FNA Based Diagnosis of Head and Neck Nodal Lymphoma

Fine-needle aspiration (FNA) biopsy has become a well established technique in the diagnosis, staging, and follow-up of patients with head and neck lesions. As in lymphoma diagnostics, FNA serves as a screening method in evaluating potentially affected lymph node for open or core biopsy. According to the World Health Organization classification of lymphoid neoplasms, today it is important to recognize cell morphology and reveal its phenotype, then combine it with different genotypic information and clinical data to provide appropriate therapy. The aim of this study was to assess the efficacy of FNA and immunocytochemistry based lymphoma diagnostic in head and neck region. We conducted a retrospective study during a period of three years where cases with either FNA diagnosis or clinical suspicion of newly recognized or relapsing lymphoma were reviewed. In the study were included patients that were referred to our laboratory from hematology department, in whom head and neck lymphadenopathia was found and lymph node FNA preceded other procedures. Two hundred eighty-five aspirations from 248 patients fulfilled study criteria. Adequate specimens were diagnosed as lymphoma in 100 cases (36%), in 65 male and 35 female patients, 76 in patients with newly discovered disease and 24 in patients with prior lymphoma diagnosis. Overall sensitivity of FNA specimens in the diagnosis of head and neck lymphomas was 90%, specificity 88%, predictive value of a positive result 97%, and predictive value of negative result 61%. Based on our results FNA corroborated with immunophenotyping by immunocytochemistry can be method of choice in primary lymphoma diagnosis as a method complementary to histopathology in lymphoma diagnostics

„THE PARIS SYSTEM (TPS) FOR REPORTING URINARY CYTOLOGY” – PRESENTATION OF DIAGNOSTIC CRITERIA IN URINARY CYTOLOGY

U ovom radu prikazujemo Parišku klasifikaciju dijagnostičkih kategorija u citologiji urina. Svjetska je radna skupina za citologiju urina osnovana 2013. godine radi uvođenja izjednačene terminologije i načina izvještavanja o nalazu u urološkoj citologiji. Smjernice za citologiju urina pod nazivom The Paris System (TPS) for Reporting Urine Cytology objavljene su 2015. godine, a uključuju specifične dijagnostičke kategorije te pouzdane citomorfološke kriterije za dijagnostiku lezija prijelaznog epitela. Dijagnostičke kategorije Pariške klasifikacije jesu: negativno na karcinom visokog stupnja (engl. Negative for High-Grade Urothelial Carcinoma – NHGUC), atipične stanice prijelaznog epitela (engl. Atypical Urothelial Cells – AUC), suspektno na karcinom prijelaznog epitela visokog stupnja (suspektno) (engl. Suspicious for High-Grade Urothelial Carcinoma – SHGUC), karcinom prijelaznog epitela visokog stupnja (engl. High-Grade Urothelial Carcinoma – HGUC), neoplazma prijelaznog epitela niskog stupnja (engl. Low-Grade Urothelial Neoplasm – LGUN) i drugi maligni primarni i metastatski tumori te različite lezije.In this paper the Paris System (TPS) of classification in diagnostic criteria for urinary cytology is presented. The International Working Group for urinary cytology was founded in 2013 with the aim to introduce consistent terminology and a standardised way of reporting findings in urine cytology. The guidelines for urinary cytology published in 2015 under the title “The Paris System (TPS) for Reporting Urinary Cytology” include specific diagnostic categories and reliable cytomorphological features in diagnosing lesions of the transitional epithelium. The diagnostic criteria of the Paris System classification are: Negative for High-Grade Urothelial Carcinoma – NHGUC, Atypical Urothelial Cells – AUC, Suspicious for High-Grade Urothelial Carcinoma – SHGUC, High-Grade Urothelial Carcinoma – HGUC, Low-Grade Urothelial Neoplasm – LGUN, other malignant primary and metastatic tumours and different lesions

FLUDARABINE, CYCLOPHOSPHAMIDE AND RITUXIMAB (FCR) IN THE TREATMENT OF CHRONIC LYMPHOCYTIC LEUKEMIA (CLL): UNIVERSITY HOSPITAL CENTRE ZAGREB EXPERIENCE

U kliničkim je studijama kombinacija fludarabina, ciklofosfamida i rituksimaba (FCR) pokazala odlične rezultate u liječenju bolesnika s kroničnom limfocitnom leukemijom (KLL) i postala zlatni standard u prvoj liniji liječenja takvih bolesnika bez znatnijih komorbiditeta. Cilj rada bio je ispitati terapijsku djelotvornost, toksičnost i provedivost ovog protokola u svakodnevnoj kliničkoj praksi. Retrospektivno su analizirani tijek i ishodi liječenja 43-oje bolesnika s KLL-om sa Zavoda za hematologiju Klinike za unutarnje bolesti Kliničkoga bolničkog centra Zagreb. Shema primjene rituksimaba razlikovala se od one u kliničkim studijama; primjenjivan je infuzijski u dozi od 375 mg/m2 u svim ciklusima, u ukupno osam doza u prvoj, odnosno šest u kasnijim linijama liječenja. Na liječenje je odgovorilo 95% bolesnika, a 83% postignulo je kompletnu remisiju. Trogodišnje preživljenje i preživljenje bez progresije bolesti u prvoj liniji liječenja (29 bolesnika) bilo je 90 i 80%, a u kasnijim linijama 86 i 62%. Teške neutropenije zabilježene su u 46% bolesnika, a teške infekcije u 9% bolesnika. Ishodi liječenja i toksični profil u svakodnevnome kliničkom radu usporedivi su s onima iz kliničkih studijaIn clinical trials the combination of fludarabine, cyclophosphamide and rituximab (FCR) demonstrated superior results and became the gold standard for first-line treatment of fit patients with chronic lymphocytic leukemia (CLL). The aim of this study was to evaluate the efficacy, toxicity and feasibility of this protocol in everyday clinical practice. We ­retrospectively analyzed the outcomes of 43 CLL patients treated at the Division of Haematology, Department of Internal Medicine, University Hospital Centre Zagreb. The dosing of rituximab differed from that in clinical trials, we administered 375mg/m2 of rituximab per cycle, in previously untreated patients for eight and in relapsed/refractory patients for six cycles. The response rate was 95% with 83% of complete remissions. Twenty–nine patients received FCR as a front–line therapy; three-year overall and progression–free survival were 90% and 80%, respectively. In relapsed/refractory disease three–year overall and progression–free survival were 86% and 62%, respectively. Severe neutropenias occurred in 46% and serious infections in 9% of patients. According to these results, the toxicity profile and treatment outcomes in everyday routine clinical practice are similar to those reported in clinical trials

Fludarabin, ciklofosfamid i rituksimab (FCR) u liječenju bolesnika s kroničnom limfocitnom leukemijom (KLL): iskustvo Kliničkoga bolničkog centra Zagreb [Fludarabine, cyclophosphamide and rituximab (FCR) in the treatment of chronic lymphocytic leukemia (CLL): University Hospital Centre Zagreb experience]

In clinical trials the combination of fludarabine, cyclophosphamide and rituximab (FCR) demonstrated superior results and became the gold standard for first-line treatment of fit patients with chronic lymphocytic leukemia (CLL). The aim of this study was to evaluate the efficacy, toxicity and feasibility of this protocol in everyday clinical practice. We ­retrospectively analyzed the outcomes of 43 CLL patients treated at the Division of Haematology, Department of Internal Medicine, University Hospital Centre Zagreb. The dosing of rituximab differed from that in clinical trials, we administered 375mg/m2 of rituximab per cycle, in previously untreated patients for eight and in relapsed/refractory patients for six cycles. The response rate was 95% with 83% of complete remissions. Twenty–nine patients received FCR as a front–line therapy; three-year overall and progression–free survival were 90% and 80%, respectively. In relapsed/refractory disease three–year overall and progression–free survival were 86% and 62%, respectively. Severe neutropenias occurred in 46% and serious infections in 9% of patients. According to these results, the toxicity profile and treatment outcomes in everyday routine clinical practice are similar to those reported in clinical trials

FLUDARABINE, CYCLOPHOSPHAMIDE AND RITUXIMAB (FCR) IN THE TREATMENT OF CHRONIC LYMPHOCYTIC LEUKEMIA (CLL): UNIVERSITY HOSPITAL CENTRE ZAGREB EXPERIENCE

U kliničkim je studijama kombinacija fludarabina, ciklofosfamida i rituksimaba (FCR) pokazala odlične rezultate u liječenju bolesnika s kroničnom limfocitnom leukemijom (KLL) i postala zlatni standard u prvoj liniji liječenja takvih bolesnika bez znatnijih komorbiditeta. Cilj rada bio je ispitati terapijsku djelotvornost, toksičnost i provedivost ovog protokola u svakodnevnoj kliničkoj praksi. Retrospektivno su analizirani tijek i ishodi liječenja 43-oje bolesnika s KLL-om sa Zavoda za hematologiju Klinike za unutarnje bolesti Kliničkoga bolničkog centra Zagreb. Shema primjene rituksimaba razlikovala se od one u kliničkim studijama; primjenjivan je infuzijski u dozi od 375 mg/m2 u svim ciklusima, u ukupno osam doza u prvoj, odnosno šest u kasnijim linijama liječenja. Na liječenje je odgovorilo 95% bolesnika, a 83% postignulo je kompletnu remisiju. Trogodišnje preživljenje i preživljenje bez progresije bolesti u prvoj liniji liječenja (29 bolesnika) bilo je 90 i 80%, a u kasnijim linijama 86 i 62%. Teške neutropenije zabilježene su u 46% bolesnika, a teške infekcije u 9% bolesnika. Ishodi liječenja i toksični profil u svakodnevnome kliničkom radu usporedivi su s onima iz kliničkih studijaIn clinical trials the combination of fludarabine, cyclophosphamide and rituximab (FCR) demonstrated superior results and became the gold standard for first-line treatment of fit patients with chronic lymphocytic leukemia (CLL). The aim of this study was to evaluate the efficacy, toxicity and feasibility of this protocol in everyday clinical practice. We ­retrospectively analyzed the outcomes of 43 CLL patients treated at the Division of Haematology, Department of Internal Medicine, University Hospital Centre Zagreb. The dosing of rituximab differed from that in clinical trials, we administered 375mg/m2 of rituximab per cycle, in previously untreated patients for eight and in relapsed/refractory patients for six cycles. The response rate was 95% with 83% of complete remissions. Twenty–nine patients received FCR as a front–line therapy; three-year overall and progression–free survival were 90% and 80%, respectively. In relapsed/refractory disease three–year overall and progression–free survival were 86% and 62%, respectively. Severe neutropenias occurred in 46% and serious infections in 9% of patients. According to these results, the toxicity profile and treatment outcomes in everyday routine clinical practice are similar to those reported in clinical trials