[INITIAL RESULTS OF DIAGNOSTIC APPLICATION OF ENDOBRONCHIAL ULTRASOUND IN CLINICAL HOSPITAL DUBRAVA]. / PRVI REZULTATI DIJAGNOSTICKE PRIMJENE ENDOBRONHALNOG ULTRAZVUKA U KLINICKOJ BOLNICI DUBRAVA.

Endobronchial ultrasound (EBUS) enables visualization of mediastinal and hilar lymph nodes as well as, in case of clinical indication, its transbronchial needle aspiration (TBNA). Simultaneous application of color doppler gives the additional prospects to investigate the blood vessels. The primary indication for EBUS is radiographic finding of mediastinal and/or hilar lymph nodes enlargement as well as the staging of non-small cell lung cancer (NSCLC). In this article we present the initial results of application of this valuable diagnostic procedure performed at the Department for pulmonary diseases in Clinical hospital Dubrava in Zagreb. According to its high sensitivity and specificity, and being safe and well tolerated, EBUS-TBNA was positioned as basic diagnostic procedure in patients with mediastinal lymphadenopathy.

Evaluation of sentinel node biopsy in oral carcinomas.

It is still a matter of debate whether sentinel node biopsy might replace neck dissection in patients with clinically negative neck lymph nodes who suffer from oral squamous cell carcinoma. In 30 patients (26 male, 4 female, average age 59.4 years) with oral squamous cell carcinoma we performed ultrasound guided punction of the lymph nodes which were lymphoscintigraphically seen together with histopatological analysis of the dissected node. Sentinel lymph node was seen in 93% cases. By use of lymphoscintigraphy sentinel node was verified in 23 patients. Ultrasound guided punction showed presence of regional disease in 10% of cases, whereas sentinel biopsy revealed 23 of the converted necks. Histopathological findings were positive in 33% of our patients. The results of this study revealed that sentinel biopsy did not reveal 27% of the patients with positive neck histopathology. In conclusion, sentinel node biopsy should be performed in selective cases as in some localizations it is easier to perform neck dissection in comparison to the sentinel node biopsy.

[Croatian Society for Clinical Cytology guidelines for thyroid cytology]. / Smjernice u citoloskoj dijagnostici stitnjace Hrvatskoga drustva za klinicku citologiju.

The main purpose of thyroid FNA (fine needle aspiration) is to separate malignant and possibly malignant nodules from benign thyroid lesions. Every patient with thyroid nodule is a candidate for FNA. Before a decision to perform an FNA, a complete history, a physical examination directed to the thyroid and cervical lymph nodes, a serum thyrotropin level, and thyroid ultrasound should be obtained. Thyroid lesion with a maximum diameter greater than 1.5 cm or nodule of any size with sonographically suspicious features is an indication for FNA. Ultrasound-guided FNA of the thyroid is recommended. The requisition form that accompanies FNA should contain the identifying data, location and size of the nodule, and relevant laboratory and clinical data. FNA diagnosis of thyroid disease is a clinicocytologic diagnosis, and correlation with clinical findings is mandatory for success. Thyroid FNA classification scheme consists of a four diagnostic categories according to the risk of malignancy: benign lesions, indeterminate lesions according to malignancy, malignant tumors, and non-diagnostic. Ancillary studies (immunocytochemistry, RT-PCR, flow cytometry) are usually helpful in borderline cases.

The importance of team work of cytologist and surgeon in preoperative diagnosis of intraoral minor salivary gland tumours.

Tumours arising from oral minor salivary glands may exhibit an overlap of clinical and morphological features that may produce diagnostic and therapeutic dilemmas. The aim of this study is to asses the value of fine needle aspiration cytology (FNAC) in differentiation of benign and malignant tumours and to render a specific diagnosis. We evaluated the team work of surgeon and cytologist to improve diagnostic accuracy. Two steps are important for accuracy: sampling aspirate that should be done together by surgeon and cytologist and cytological microscopic analysis of the smears that should be performed by an experienced cytologist. The study included 132 patients with intraoral minor salivary gland tumours between 2002 and 2011. Adequate material was obtained from 121 (91.7%) patients. FNAC was usually performed by cytologist in a team with maxillofacial surgeon at cytology department that is more convenient for preparing the samples and especially for ROSE procedure (rapid-on site evaluation) of smears. In such a way the cytologist checked the adequacy of samples and decided whether some ancillary techniques should be used and therefore repeat FNAC. A total of 82 patients underwent surgery, 40 with malignant and 42 with benign tumours. Preoperative cytological diagnoses were compared with histopathological ones using histopathology as a gold standard. The most common benign tumour was pleomorphic adenoma and among malignant tumours adenoid cystic carcinoma. The most commonly affected site was the palate. The team work of surgeon and cytologist achieved specificity of 95.1%, sensitivity of 97.6% and diagnostic accuracy of 96.3%. We can conclude that although subclassification of some tumour types of salivary glands remains poor, FNAC is invaluable in patient triage and therefore should be considered in the first line investigations of these lesions by the cytologist and surgeon.

Diffuse sclerosing variant of thyroid carcinoma presenting as Hashimoto thyroiditis: a case report.

The aim of report is to present a case of a rare diffuse sclerosing variant of a papillary thyroid carcinoma. A 15-year old girl referred for ultrasound examination because of painless thyroid swelling lasting 10 days before. An ultrasound of the neck showed diffusely changed thyroid parenchyma, without nodes, looking as lymphocytic thyroiditis Hashimoto at first, but with snow-storm appearance, predominantly in the right lobe. Positive thyroid peroxidase antibodies (TPO-AT) also suggested Hashimoto thyroiditis. Repeated US-FNAB (fine needle-aspiration biopsy) of the right lobe revealed diffuse sclerosing variant of papillary thyroid carcinoma and patient underwent total thyreoidectomy. Patohistologic finding confirmed diffuse sclerosing variant of a papillary thyroid carcinoma in the both thyroid lobes and several metastatic lymph nodes. Two months later patient recived radioablative therapy with 3700 MBq (100 mCi) of 1-131 followed by levothyroxine replacement. At the moment, patient is without evidence of local or distant metastases and next regular control is scheduled in 6 months. In conclusion, a diffuse sclerosing variant is rare form of papillary thyroid carcinoma that echographically looks similar to Hashimoto thyroiditis and sometimes could be easily overlooked.

[Trends in stage and age characteristics of the female breast cancer treated in 11-year period at University Hospital "Dubrava"]. / Analiza dobne distribucije i stupnja lokalne prosirenosti raka dojke kod bolesnica operiranih u 11-godisnjem razdoblju u Klinickoj bolnici "Dubrava".

In a retrospective study we analyzed and compared the factors of local spread of breast cancer, namely tumor size and status of regional lymph nodes and patient age in the period from 1997 to 2007. The study includes only those patients in whom the diagnosis was made in an open surgical biopsy and it was possible to determine the local extent of disease at the time of diagnosis (pT and pN stage), a total of 1202 patients. We also compared the findings in patients younger and older than 50 years. The average tumor size at diagnosis was 2.2 cm and ranged between 1.7 cm (2005) and 2.4 cm (1997, 1999). The percentage of metastases in regional lymph nodes varied between 37% (2001) and 50% (2000 and 2006). The average of 6% of all cases of breast cancer was diagnosed in non-invasive (in situ) stage. The mean patient age was 56.8 years and for the observed age ranged between 54.3 (1999) and 59.1 (2004) years. There was no statistically significant difference in relation to disease stage or patient age at diagnosis observed throughout the entire period of time. There was also no statistical difference in stage of disease at the time of diagnosis in patients younger and older than 50 years.

Schwannoma of the tongue in a child.

A schwannoma or neurilemmoma is a benign, slow growing, usually solitary and encapsulated tumour originating from Schwann cells of the nerve sheath. Approximately 25-40% of all schwannomas are seen in the soft tissues of the head and neck, often originate from the acoustic nerve. Intraoral schwannomas are rare and account for 1% of schwannomas of the head and neck region. We report the case of a 10-year-old boy diagnosed with a schwannoma of the tongue. The purpose of this report is to emphasize the possibility of diagnosing schwannoma among all other lingual lesions in children. The disease itself was diagnosed histologically after complete surgical excision. Five years after surgical treatment, the patient is without signs of recurrence. This paper highlights the importance of a multidisciplinary approach in the diagnosis and surgical treatment of this very rare entity.

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.

Fine needle aspiration cytology of chondroid syringoma.

Chondroid syringoma (CS) is a rare, benign, appendageal tumor with diagnostic feature of myxochondroid stroma supporting eccrine and apocrine epithelial structures. The usual presentation is a painless, slowly growing mass, typically located in the head and neck region. It usually affects middle-aged and older male patients. Because of its unremarkable clinical presentation it is often overlooked. It should be included in the differential diagnosis of cutaneous head and neck tumors, especially in middle-aged men. Optimal treatment of CS is total surgical excision. We present a 63 year-old man with a small nodule on the neck with 5 years of duration. The diagnosis was made initially on fine needle aspiration cytology that was performed by ultrasound guidance and confirmed subsequently by histology. FNA cytology may be very useful to determine diagnosis before excision.

Fine needle aspiration cytology of metastatic Merkel cell carcinoma.

Merkel cell carcinoma (MCC) is uncommon cutaneous malignant neuroendocrine tumour of the elderly people with rapidly growing skin nodules found frequently on sun-exposed areas of the body. MCC is often an aggressive tumour with high tendency for local recurrence, lymph node involvement and distant metastases. This paper reports a case of metastatic MCC diagnosed by fine needle aspiration cytology (FNAC), flow cytometric deoxiribonucleated acid (DNA) analysis, pathohistology and electron microscopy. The cytological features in aspirate (stained with Papenheim and Papanicolaou staining) included increased cellularity, discohesive groups of small-to-medium size malignant cells with uniform, round-to-oval nuclei with moulding effect, fine chromatin, multiple micronucleoli and scanty cytoplasm. DNA flow cytometric analysis of the aspirate showed unexpected results for clinically aggressive behaviour of this tumour (the patient died after 21 months), and revealed that tumour contained diploid peak with DNA index of 1.1. The proliferation was high with elevated S-phase fraction (21%). The cytological diagnosis of possible metastatic MCC was confirmed by histological one as well as by electron microscopy presented the pathognomonic features for this tumour: dense-core neurosecretory granules with diameter of 100-250 nm surrounded by whorls of intermediate filaments. MCC provides an enormous challenge for the morphologist because of a wide range of differential diagnosis and for the clinician because this tumour has a highly malignant potential for local recurrence, nodal and distant spread and very often is combined with other tumours. Therefore it is important to perform FNAC of different lesions in the same patient because it can distinguish MCC from the other tumours.

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.

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.

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.

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 in diagnosing rare breast carcinoma--two case reports.

In this paper, we are presenting the two cases of very rare tumors: breast sebaceous carcinoma, which has been described for the first time in Croatian medical literature, and pure breast squamous carcinoma. First case, sebaceous carcinoma, is still quite unknown regarding its morphological characteristics and biological behavior. In the second case, squamous carcinoma, also very rare, was found in a patient who previously had a number of diagnosed head and neck skin carcinomas, and was diagnosed as primary squamous breast carcinoma. As a first case we present a 85-year-old female with a two months history of swelling of the left breast under the mammilla. The second one, a 69-year-old female presented to our hospital in January 2008 with a two months history of a lump in the lower outer region of the left breast and enlarged lymph nodes in left axillary region. Fine needle aspiration cytology (FNAC) of the breast was performed in order to diagnose the exact type of both tumours. This methodology was found important in diagnosis, but in both cases showed certain limitations in diagnosing such rare tumors. The final diagnoses were determined after carefully synthesizing the histological findings and clinical data. Careful and accurate classification of these tumors is necessary. A detailed analysis of their biological behavior and response to the therapy is necessary in order to formulate definite recommendations in managing these patients/diseases.

Monocytes in metabolic disorders--opportunities for flow cytometry contributions.

Chronic inflammation has arised as a major underlying cause of atherosclerosis, obesity and diabetes. It is mediated by cells of innate immune system like macrophages but also by their antecedents, circulating monocytes. Roles of monocyte subsets and different markers of monocyte activation in the context of metabolic disorders have been reviewed. Applying cell based approach through flow cytometry in this field has resulted with new understanding of pathophysiologic mechanisms. Possible implications of these insights in diagnosis, prognosis and revealing of therapeutic targets in metabolic disorders remain a challenge for future.