[GUIDELINES FOR DIAGNOSIS AND TREATMENT OF PATIENTS WITH MYELODYSPLASTIC SYNDROME].

Myelodysplastic syndrome Working Group of the Croatian Cooperative Group for Hematologic Diseases (CROHEM), Referral center of the Ministry of Health of the Republic of Croatia for diagnostics and treatment of MDS, as well as the Croatian Society for Haematology of the Croatian Medical Association have made Croatian guidelines for diagnosis and treatment of myelodysplastic syndrome (MDS). MDS is a heterogeneous group of clonal hematopoietic stem cell disorders characterized by ineffective hematopoiesis, dysplasia, cytopenia and risk of transformation to acute myeloid leukemia (AML). Diagnosis is based on morphological characteristics of hematopoietic cells supplemented with the cytogenetic analysis and bone marrow flow cytometry. Due to great differences in the natural course of the disease, i.e. time to progression to AML and the expected time of survival several scoring systems have been developed to determine the disease risk. The treatment of patients with MDS is based on the risk factors of the disease as well as the individual risk of treatment.

Biliary brush cytology for the diagnosis of malignancy: a single center experience.

Differentiation between benign and malignant biliary strictures is critical to the provision of adequate treatment. Brush cytology during the endoscopic retrograde cholangiopancreatography (ERCP) is the most commonly used method for obtaining tissue confirmation of the nature of biliary strictures. It's specificity is remarkably high but reported sensitivities for the diagnosis of malignancy are low. Aim of our study was to assess sensitivity and specificity of biliary brush cytology in our institution, to find out main causes of false negative diagnoses and to confirm impression that the team approach has impact on sensitivity. Gold standard for diagnosis was definitive surgical histology or adequate clinical follow up for minimum of six month. Direct smears made by cytotechnician at the endoscopy room, and stained according to Papanicolaou and May-Grünwald Giemsa (MGG) were examined for well-recognized features of malignancy on conventional smears as a part of diagnostic routine. Cytologic diagnoses were benign, atypical/reactive, suspicious for malignancy and malignant. Of 143 brushings with available definitive diagnosis 36 (25%) had malignant cytologic diagnosis and 91(63.6%) were classified as benign, 3 were atypical/reactive and 13 suspicious for malignancy with 20 "false-negative" cases. When specimens with atypical and suspicious cytology were excluded from data analysis sensitivity was 64% and specificity was 100% and when suspicious findings were taken into account as true positives sensitivity rose to 71%. We find that biliary brush cytology, although mainly depending on the skill of endoscopist, as well as the experience of the cytologist, is a valuable method for obtaining accurate tissue diagnosis of biliary strictures, thus solving eternal diagnostic dilemma: benign or malignant.

Flow cytometric analysis of deep-seated lymph nodes.

Flow cytometry (FC) immunophenotyping is an important tool in the evaluation of lymphadenopathy and is widely used in the diagnosis of non-Hodgkin's lymphomas (NHLs) on fine-needle aspirates of lymph nodes and extranodal sites. Because at least 80% of NHLs are of B-cell type, detection of immunoglobulin (Ig) light-chain-restriction is the most commonly used method for confirmation of monoclonality. The aim of our study was to evaluate usefulness of endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) for FC analysis from deep-seated lymph nodes and to compare results of FC clonality analysis to cytomorphologic diagnosis of sampled lymph nodes. For cytological diagnosis direct smears were made, selected slide was stained for rapid-on site evaluation procedure. Sixteen patients with suspected NHL of deep-seated lymph nodes obtained by EUS-FNA were submitted for FC clonality analysis using four-color multiparameter flow cytometry stained with kappa/lambda/CD19/CD45. Clonality analysis was performed on 11 samples. Monoclonality was demonstrated in seven of 11 cases cytologically diagnosed as NHL and four of 11 cases cytologically diagnosed as benign were polyclonal. Our results show that EUS-FNAC with FC is a sensitive and specific tool in the diagnosis of deep-seated B-NHL. Cytologic diagnosis combined with FC clonality analysis can be performed in majority of cases and may eliminate need for open biopsy in some cases.

Intraoperative imprint cytological assessment of the subareolar tissue of the nipple areola complex (NAC).

One of the criteria of selection for skin sparing mastectomy (SSM) with nipple areola complex (NAC) preservation is to exclude the neoplastic involvement of subareolar tissue (NAC base) in order to minimize the possibility of local recurrence. The most common way to assess the possible neoplastic involvement is intraoperative frozen section of the NAC base tissue. Because of its limitations, particularly the false negative results due to unsampling, we tried to use intraoperative imprint cytology for more thorough intraoperative assessment. The aim was to compare intraoperative imprint findings with the definitive histology of the NAC base, to evaluate diagnostic accuracy of this method and possibility to substitute frozen section for intraoperative assessment of NAC base. A prospective clinical study was conducted of 208 consecutive female patients who underwent open biopsy because of carcinoma. Intraoperative imprints were taken from the excised subareolar tissue which was then routinely processed for definitive histology. Imprint findings designated positive, negative, suspicious or atypia, were compared with definitive histological findings. Our results with 7.5% false negative rate, 9.8% false positive rate, sensitivity of 50% and specificity of 87.58% argue that imprint cytology might not be sufficient as an exclusive method for the intraoperative assessment of the NAC base though it should be used routinely in conjunction with frozen section examination.

Fine-needle aspiration cytology of apocrine hidradenoma.

An apocrine hidradenoma is a benign adnexal neoplasm, usually covered by intact skin, but may show superficial ulceration and serous discharge. This feature is raising the possibility of malignancy as it was in our case of macroscopically suspicious tumour. We described cytomorphologic features of cutaneous nodule that might be a lead to the cytologic diagnosis of hidradenoma, but primary or secondary malignant tumour has been ruled out first.

Clinical cytology and primary health care of children and adults.

Clinical cytology is a diagnostic branch of medicine, best known by the Papa test in gynaecology. But, cytology can be applied in almost all fields of clinical medicine. Its advantages--high accuracy, simplicity, with little or no aggressiveness and low cost--are not used as widely as they could be. Medical practice, as well as medical research, and also medical education, are nowadays often directed at profitable use and not at the real benefit of the patient. Primary practitioners do not have enough chance to get acquainted with clinical cytology as a whole although they need true information, based on the cost-effectiveness and patient-benefit. A panel discussion on this subject was organised at the 4th Croatian Congress of Clinical Cytology, in Split, October 11-14, 2009 by the Croatian Society for Clinical Cytology-Croatian Medical Association, to inform primary practitioners about the possibilities of cytodiagnostics in the health care of children and adults. Indications for cytodiagnostics in infectious diseases (T. Jeren and A. Vince), haematology (I. Kardum-Skelin), pulmonology (S. Smojver-Jezek), thyroid diseases (A. Knezevic-Obad), breast diseases (I. Kardum-Skelin), gastroenterology and urology (G. Kaic) were discussed, as well as technical procedures and the interpretation of the cytological findings. Moderator (Z. Znidarcic) opened the panel with presentation about the role of clinical cytology, particularly in the primary health care. The discussion finally pointed at the necessity of better communication between primary practitioners and cytologists. This review article presents contents of the panel discussion.

The importance of urgent cytological examination of synovial fluids in differentiation inflammatory and non-inflammatory joint diseases.

The aim of this study was to imply the possibilities of the urgent cytological examination of synovial fluids in differential diagnosis of arthropathies and to motivate the clinicians to use this method. It gave valuable information particularly with respect to differentiate the inflammatory and non-inflammatory joint diseases. This study included 115 synovial fluids obtained by fine needle aspiration (FNA) of the swollen knee from the patients in the period between 2003 and 2008. At our department the urgent cytological examination of the synovial fluids consisted of macroscopic analysis that includes volume, colour, clarity, viscosity and mucin clot test, native microscopic analysis for crystals and tissue fragments, counting the total nucleated cell count and semiquantitative microscopic analysis for neutrophil granulocyte percentage on the slides stained with Hemacolor rapid staining. All cytological analyses were done within one hour since FNA. According to our results the clarity, viscosity, mucin clot test, the total nucleated cell count and the neutrophil granulocyte percentage enabled distinction between inflammatory and non-inflammatory diseases with statistically significant difference at the 0.01 level but we could not differentiate these two groups of illnesses according to volume and colour. In inflammation the total nucleated cell count and the neutrophil granulocyte percentage was greater than in non-inflammation, the clarity was only translucent and opaque, the viscosity was low and the mucin clot test was negative. In non-inflammatory diseases the clarity varied from transparent to opaque, the total nucleated cell count and the neutrophil granulocyte percentage was smaller than in inflammatory diseases, the viscosity was high and consequently the mucin test was highly positive in all samples. Crystals were detected in only 12 samples of synovial fluids, mostly in inflammation and they were all monosodium urate (MUS) so we could diagnose gout. We could conclude that the urgent cytological analysis of the synovial fluid is a very useful, simple and reliable basic diagnostic screening test in differentiation inflammatory and non-inflammatory joint diseases and we recommended using it as the initial test in the diagnostic procedure of these illnesses using our protocol.

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.

Misleading presentations of malignant breast diseases--role of clinical cytology.

We described two examples with misleading presentations to draw attention to the role of clinical cytology as apart of multidisciplinary approach to breast lesions. In the first case--Paget's disease of the nipple, there was no obvious clinical and radiological evidence of breast cancer, while the second case--primary non-Hodgkin lymphoma of the breast imitated advanced breast carcinoma. The question is whether accurate and fast diagnoses can be made without cytological examinations. It must be kept in mind that first-hand clinical information and contact with the patient is necessary in rendering accurate cytological diagnoses.

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.

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.

Hydronephrotic Kidney Previously Treated for Tuberculosis: Rare Primary Squamous Cell Carcinoma of Renal Pelvis Diagnosed by Fine-needle Aspiration Cytology.

Primary squamous cell carcinoma (SCC) of the renal pelvis is a very rare tumor often associated with renal calculi and chronic infections. There are only a few articles in literature which report renal pelvis SCC in kidneys treated for renal tuberculosis, diagnosed after nephrectomy. We report the case of SCC in a hydronephrotic kidney previously treated for tuberculosis, diagnosed by ultrasound (US)-guided fine-needle aspiration cytology (FNAC), prior to core biopsy and nephrectomy. Our report highlights the utility of FNAC and the need for a careful search for renal collecting system tumors, in patients previously treated for renal tuberculosis.

Alpha-methylacyl-CoA racemase (AMACR) in fine-needle aspiration specimens of prostate lesions.

The elevated expression of P504S gene and its product alpha-methylacyl-CoA racemase (AMACR) can serve as a molecular marker for prostate cancer. The goal of this study is to investigate P504S/AMACR expression in fine-needle aspiration smears and correlate it with cytological diagnosis. Immunocytochemistry was performed in 35 patients with morphological diagnosis of prostate carcinoma (n = 16), atypia (n = 15), and benign hyperplasia (n = 4). Among 16 malignant cases there were two low-grade, eight intermediate, and six high-grade prostate carcinomas. Cytoplasmic positivity is analyzed qualitatively as predominantly diffuse or focal and quantitatively as <5%, 5-50%, and >50% of cells. Benign cases showed no P504S/AMACR expression. Positive staining was recorded in 75% of malignant cases, but in the majority of them it was weak and focal or diffuse and in a small amount of cells. The most intensive staining was seen in low-grade carcinomas and some atypical cases. This observation indicates a correlation between P504S/AMACR expression and differentiation of cells. P504S/AMACR staining might be of great value in cytodiagnosis of prostate lesions as well as an example of the characterization of cells at the molecular level using fresh tissue obtained by fine-needle aspiration.