BETA-CATENIN EXPRESSION IN MALIGNANT MELANOMA

 Martina Žigmund1, Nives Pećina-Šlaus1, 2, Vesna Kušec3, Tamara Nikuševa–Martić1, 2, Mirjana Čačić4, Mario Šlaus5, Floriana Bulić-Jakuš1

 1Department of Biology, School of Medicine University of Zagreb, Zagreb, Croatia, 2Croatian Institute for brain research, School of Medicine University of Zagreb, Zagreb, Croatia, 3Clinical Institute of Laboratory Diagnostics, University Hospital Centre Zagreb, Zagreb, Croatia, 4Department of pathology, Clinical Hospital Centre Zagreb, Zagreb, Croatia, 5Department of Forensic Medicine, School of Medicine University of Zagreb, Zagreb, Croatia.

 Beta-catenin is bound to E-cadherin in adherence junction formation, but also functions as a signaling molecule in the WNT pathway. We investigated beta-catenin expression in 41 superficial spreading melanomas. Our melanoma sample was analyzed by immunohistochemistry and evaluated by image analysis as staining density, i.e. light permeability (LP). In normal skin beta-catenin showed homogenous membranous staining of the epidermis. Comparison of relative LP for beta-catenin in tumor tissue and adjacent skin did not show any differences (157.8 compared to 156.6; t = -1.087, P0.283). However, the cellular location of beta-catenin changed considerably in the melanoma. The protein was observed in the cytoplasm in 32% of patients, in 29% in the cell membrane, in 24% in both the cytoplasm and membrane, in 5% in the cytoplasm and nucleus, while in 9.8% of patients beta-catenin was not observed. There was a marked difference in beta-catenin distribution correlated to the Clark stages of melanoma progression. Patients with Clark 4 and 5 had significantly less beta-catenin compared to patients with Clark 2 and 3 (P=0.01).

Our results suggest that changes of beta-catenin levels have a role in melanoma and could be used as markers of disease progression.

 

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PP02

FINE NEEDLE ASPRATION CYTOLOGY (FNAC) IN A CASE OF POROCARCINOMA

 E. Garcia-Ureta, E. Carro Rey, O. Robles Viega, R. Alvarez Rodriguez  

Citologia Servicio De Anatomía Patológica Hospital Universitario Juan Canalejo, La Coruña; Spain

             INTRODUCTION: Porocarcinoma is a very rare malignant tumor of the eccrine sweat gland. Just a few porocarcinoma have been studied by FNAC. FNAC findings of a case of porocarcinoma are reported.

            CASE REPORT: A 63 year-old woman presented with a painless non ulcerated nodule located in the parotid gland. She underwent surgery, two months earlier, and removed a lesion of 2 cm on the same cheek. FNAC of the mass was performed according to the standard procedure, using a 23 gauge needle. Smears were prepared from FNAC of the lump, the slides were routinely processed (MGG, Pap).

            RESULTS OF CYTOLOGICAL FINDINGS: The aspirates were cellular and consisted of aggregates of cells occasionally with duct differentiation. Two types of cells were found, one had abundant cytoplasm, some of them with intracytoplasmatic lumina, in which small to large nuclei; with small nucleoli, chromatin was finely granular. Another type of cell had scanty cytoplasm and a round to oval nucleus with small nucleoli.

The patient underwent surgery and histopathological analysis revealed an enlarged lymph node with a metastatic process caused by a porocarcinoma. The previous biopsy of the cheek was revised and the diagnosis of porocarcinoma was made.

            CONCLUSIONS: The cytological features in this case was representative with presence of cuticular and poroid cells. The main difficulty was not to think of this process. The differential diagnosis included other eccrine sweat gland tumors.

 

 

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PP03

FREQUENCY OF LENTIGO MALIGNA MELANOMA IN TWO CROATIAN REGIONS

 A. Krvavica1, M. Vučić2, M. Ulamec2, C. Lež, 3 M. Belicza2

1Department of Pathology, General Hospital Zadar, Croatia; 2Ljudevit Jurak Department of Pathology, Sestre milosrdnice University Hospital, Zagreb, Croatia; 3Department of Pathology, General Hospital Zabok, Croatia

 
AIM OF THE STUDY: Lentigo maligna melanoma accounts for 5 to 10% of all primary skin melanomas and occurs mainly in the elderly; it is an invasive melanoma that arises from preinvasive lesion lentigo maligna confined within the epidermis. Sir John Hutchinson first described lentigo maligna in 1890; the disease continues to be called Hutchinson melanotic freckle on occasion. The aim of our study was to analyze lentigo maligna melanoma subtype during a 7-year period (1999-2005) among the biopsy specimens in the General Hospital Zadar and Sestre milosrdnice University Hospital Zagreb.

MATERIALS AND METHODS: We used data bases from the Department of Pathology, General Hospital Zadar and the computerized Tumor Registry Thanatos from Ljudevit Jurak University Department of Pathology, Sestre milosrdnice University Hospital Zagreb. We compared clinical parameters (sex, age and localization) and histological parameters (Clark, Breslow level of invasion, and pT classifications) of lentigo maligna melanoma between the two climatically different regions.

RESULTS: Lentigo maligna and lentigo maligna melanoma accounts for 7.9 % of all diagnosed primary skin melanomas in Zadar and for 8% in Zagreb. Generally, patients with lentigo maligna melanoma are older than 40 years. Mean age at time of diagnosis of lentigo maligna melanoma in Zadar was 61.3 years and in Zagreb 62.5 years (in both hospitals average age for all types of primary skin melanomas was 59.2 years). Preinvasive lesion, lentigo maligna was diagnosed more frequently in Zagreb with mean age of 68.5 years in both hospitals. F/M ratio was 2.3/1 amongst Zadar patients compared to 2.8/1 amongst Zagreb patients. Lentigo maligna melanoma most commonly affects the sun-exposed skin of the head and neck, with a predilection for the nose and cheek and skin of lower extremities. Clark/ Breslow stages and pT classification among the patients in both Hospitals were distributed equally (in majority of the cases Clark and Breslow stage II and pT1).

DISSCUSION AND CONCLUSION: The overall incidence of cutaneous melanoma is increasing faster than any other neoplasm. Increased exposure to ultraviolet radiation and long-term cumulative ultraviolet injury has been a major risk factor for lentigo maligna melanoma. The incidence of lentigo maligna and lentigo maligna melanoma increases in sunnier climates. They often present with a prolonged phase of slow growth but once the invasion has occurred the prognostic features are identical to all other melanoma subtypes. In our study distribution of analyzed parameters of lentigo maligna melanoma subtype were similar for costal and continental part; except for more frequently diagnosed preinvasive lesions of lentigo maligna in the Zagreb hospital. Many patients with lentigo maligna melanoma have a history of multiple actinic lesions and non-melanoma skin cancers.  Education plays an integral role in earlier recognition and follow-up care of patients with lentigo maligna melanoma.

References

1. STEVENSON O, AHMED I. Am J. Clin Dermatol 2005;6:151-64.
2. WHITEMAN DC, WATT P, PURDIE DM et al. J Natl Cancer Inst 2003;95:1801-2.
3. SWETTER SM, BOLDRICK JC, JUNG SY et al. J Invest Dermatol 2005;125:685-91.
4. DURNICK A, STOLZ W, LANDTHALER M, VOGT T. Dermatol Surg 2004;30:813-6.

 

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PP04

BALANITIS CIRCUMSCRIPTA PLASMACELLULARIS

 Z. Paštar1, J. Radoš2, J. Lipozenčić2, M. Skerlev2, D. Lončarić2

 1Health Department, Ministry of Defense, Zagreb, Croatia; 2Department of Dermatology and Venereology, Zagreb University Hospital Center and School of Medicine, Zagreb, Croatia

 
DEFINITION: Balanitis circumscripta plasmacellularis (BCP) is an idiopathic, benign condition of the elderly uncircumcised men genitalia1, 2, 3. Etiology is unknown. It is presented as sharply demarcated bright red glistering patch on the glans and prepuce. Analogous lesions of the female genitalia are known as vulvitis plasmacellularis.

OBJECTIVE: Our purpose is to discuss histological patterns in BCP that are seen in our patients.

Sequence of histopathologic changes is compatible with the thesis that balanitis of Zoon results from irritation or mild trauma affecting barely keratinized skin in a moist environment.

The earliest histopathological changes are thickening of the epidermis, parakeratosis and a patchy lichenoid infiltrate of lymphocytes and some plasma cells.  More advanced cases show atrophy of the epidermis, superficial erosions, scattering of neutrophils in the upper reaches of the epidermis, scant spongiosis, extravasation of erythrocytes and a much denser infiltrate with many plasma cells.  At even later stages additional findings may be: subepidermal clefts, sometimes with loss of the entire epidermis, marked fibrosis of the superficial dermis, and many siderophages. Plasma cells usually exceed 50% of the present cells, although they can be assessed as low as few, moderately dense to dense infiltrate, but they could always be detected.2 Lymphocytes are constantly present, polymorphonuclear leucocytes are never predominant while eosinophils are sparse2.

CONCLUSION: Disorders that clinically mimic BCP are numerous and recognition of histopathologic features of BCP allows differentiation from premalignant, infective and other inflammatory penile lesions which may be more responsive to treatment.

 References:

1  . DAVIS DA, COHEN PR.  Balanitis circumscripta plasmacellularis.  J Urol 1995;153:424-6.

2  . WEYERS W, ENDE Y, SCHALLA W, DIAZ-CASCAJO C.  Balanitis of Zoon: a clinicopathologic study of 45 cases.  American Journal of Dermatopathology 2002;24:459-67.

3    JOHNSON RA. Diseases and Disorders of the Male Genitalia.  In:Freedberg IM, Eisen AZ, Wolff K, Austen KF, Goldsmith LA, Katz SI, editors. Fitzpatrick`s Dermatology in General Medicine, 6th ed. McGraw Hill, New York (NY), 2003:1091-1107.

 

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PP05

LOFGREN’S SYNDROME PRESENTED WITH ERYTHEMA NODOSUM LIKE ERUPTION – A CASE REPORT

 J. Radoš

 University Department of Dermatology and Venereology, Zagreb University Hospital Center, Zagreb, Croatia

 
BACKGROUND: Sarcoidosis is a multisystemic granulomatous disease of unknown etiology that has a wide variety of clinical manifestations. Cutaneous involvement is divided into specific categories with histology of non-caseous epitheliod granulomas and non-specific categories which do not show granulomas, the most common being erythema nodosum. Löfgren’s syndrome, which is defined as the association of erythema nodosum or periarticular ankle inflammation with unilateral or bilateral hilar or right paratracheal lymphadenopathy, is an acute and usually self limited form of sarcoidosis.

OBJECTIVE: To report a very uncommon histological pattern in acute sarcoidosis.

CASE REPORT: A 57-year old women with an explosive onset of erythematous, subcutaneous nodules on calves and maculopapular eruption on the dorsal side of the feet with periarticular ankle inflammation was hospitalized at our Department in March 2006. She was febrile, with malaise and keratoconjunctivitis, without respiratory difficulties. Laboratory test revealed high erythrocyte sedimentation rate, pathological liver test, and elevated serum angiotensin converting enzyme. A chest X-ray revealed bilateral hilar lymphadenopathy. Histopathological findings from erythema nodosum like lesion showed circumscribed granulomas of epitheloid cells through the dermis with extension into subcutaneous tissue. The histology of maculopapular lesion showed discrete, round to oval granulomas in superficial dermis. Other causes of granulomatous inflammation were excluded.

CONCLUSIONS:While erythema nodosum is the most common non-specific skin lesion of sarcoidosis, the erythema nodosum like eruption in our case was a specific sarcoid lesion. Cutaneous sarcoidal granulomas appear to be associated to poorer prognosis and increased incidence of pulmonary fibrosis and uveitis. This patient should be followed up carefully.

References

1. OKAMOTO H, et al. Erythema nodosum-like eruption in sarcoidosis. Clini and Exp Dermatol 1994;19:507-510.
2. MANA J, et al. Lofgren’s syndrome revisited: a study of 186 patients. Am J Med. 1999;107:240-245.
3. AHMED I, et al. Subcutaneous sarcoidosis: Is it specific subset of cutaneous sarcoidosis frequently associated with systemic disease? J Am Acad Dermatol 2006;54:55-60.

 

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PP06

GLIOBLASTOMAS EXHIBIT LOSS OF HETEROZYGOSITY OF THE TUMOR    SUPRESSOR GENE APC

Nives Pećina-Šlaus1, 3, Vili Beroš2 , Tamara Nikuševa-Martić1, 3, Floriana Bulić-Jakuš3

 1Laboratory of Neurooncology, Croatian Institute for Brain Research, School of Medicine University of Zagreb, Zagreb, Croatia; 2Department of Neurosurgery, Sestre milosrdnice University Hospital, Zagreb, Croatia;3Department of Biology, School of Medicine, University of Zagreb, Zagreb, Croatia

This study analyzes tumor suppressor gene – adenomatous polyposis coli (APC) in 28 patients with glioblastoma, the most aggressive astrocytic form. APC protein has structural role in adherens junctions, but also plays a signaling role as a negative regulator of the WNT pathway. The etiology and pathogenesis of tumors of the central nervous system are still inadequately explained. Our interest in APC gene stemmed principally from the findings that wild-type APC protein is highly expressed in the central nervous system, and upon the finding that it is critically involved in particular syndromes, among which the brain tumors play a significant role. Glioblastoma samples were tested for gene instability by PCR/loss of heterozygosity using RFLP method. Two polymorphic markers were used: an Rsa I polymorphic site in exon 11, and an Msp I polymorphic site in exon 15. The results of our analysis for both markers showed allelic loss of the APC gene in 33.3% of our sample out of 21 heterozygous patients (heterozygosity 75%). Another 23.8% of samples demonstrated allelic imbalance of the APC allele in tumor tissue. Altogether, there were 12 samples (57%) demonstrating instability of this tumor suppressor gene. Despite increasing knowledge on glioma biology and genetics, the prognostic tools for glioblastoma still need improvement. Our findings on genomic instability of APC gene may contribute to better understanding of glioblastoma genetic profile and could be used as a prognostic marker of disease evolution and progression.

  

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PP07

DISTRIBUTION OF TENASCIN-C IN GLIOBLASTOMA – ITS RELEVANCE TO TUMOR CELLS PROLIFERATION AND PATIENT SURVIVAL

 S. Behrem, K. Žarković¹, N. Eškinja², N. Jonjić

 Department of Pathology, Rijeka University School of Medicine, Rijeka, Croatia; ¹Department of Pathology, University Hospital Center Zagreb, Zagreb, Croatia; ²Department of Neurosurgery, University Hospital Center Rijeka, Rijeka, Croatia

AIM: To analyze the distribution pattern of tenascin-C, an extracellular matrix protein, in glioblastoma and to investigate its association with tumor cells proliferation and patient survival.

 METHODS: Forty-six cases of glioblastoma were analyzed in a retrospective study. All cases were immunohistochemically stained for tenascin-C and MIB-1 (Ki-67). The proliferation index was expressed as a percentage of Ki-67 positive cells. Survival probabilities were computed according to the Kaplan-Meier method.

 RESULTS: Tenascin-C showed the focal and the diffuse expression in the intercellular space. Focal distribution was found in 66% of the tumors and the diffuse one in 34% of the cases. In 10 cases diffuse tenascin expression was found in the peritumoral brain tissue. The proliferative index was higher in tumors with diffuse tenascin-C expression (median 31%, range 8.96-53.63%), in comparison to those with focal tenascin-C expression (median 20%, range 5.17-46.47%). This difference was significant (p=0.008). But, the survival rate was significantly higher in patients with diffuse, rather than focal tenascin-C expression (p=0.039).

 CONCLUSION: Proliferative activity is associated with a distribution pattern of tenascin-C in glioblastoma. Moreover, the pattern of localization is also associated with the survival rate. Higher survival rate in patients with diffuse intercellular tenascin-C expression, in spite of higher proliferative index, and finding the diffuse tenascin-C expression in peritumoral tissue, may indicate some protective role of tenascin-C in tumor progression.

 References

1.  LEINS A et al. Expression of tenascin in various human brain tumors and its relevance for survival in patients    with astrocytoma. Cancer 2003;98:2430.

2.  KIM et al. Expression of tenascin-C in astrocytic tumors: its relevance to proliferation and angiogenesis. Surg   Neurol 2000;54:235.

3.  EMOTO K. et al. Annexin II overexpression correlates with stromal tenascin-C overexpression: a prognostic marker in colorectal carcinoma. Cancer 2001;92:1419.

  

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PP08

MENINGEAL INVOLVEMENT IN MULTIPLE MYELOMA (MM) – REPORT OF TWO CASES WITH CYTOLOGIC AND IMMUNOCYTOCHEMICAL DIAGNOSIS

 E. Garcia-Ureta, E. Carro Rey, O. Robles Viega, R. Alvarez Rodriguez

 Citologia Servicio De Anatomía Patológica Hospital Universitario Juan Canalejo, La Coruña, Spain

  INTRODUCTION: Although neurological manifestations often complicate the course of patients with multiple myeloma (MM) the invasion of the SCN is very rare, with few reported cases in the literature. We describe cytological cerebrospinal fluid (CSF) findings in two patients with MM who presented with meningeal myelomatosis.

            CASE 1: A 47 year-old man with MM with several episodes of neurological manifestations.

            CASE 2: A 53 year-old woman with MM with several hospital admissions, sometimes with neurological symptoms.

            MATERIAL AND METHODS: The CSF was obtained by lumbar needle biopsy and valued by cytology for cellularity and differential cell count. Smears were prepared from sediment obtained by cytocentrifugation and stained with MGG and Pap and ICQ.

            RESULTS: Cytological findings are summarized in the following table:

 

		C ml	H ml	DIFFERENTIAL COUNT
				P C	L	M	N	E
CASE I	I	38	16	72	18	6	4
	II	117	32	79	13	7	1
CASE II	I	79	210	83	7	5	4	1
	II	43	85	93	2	4	1
	III	142	176	87	3	4	6

 PC: plasma cells; L: lymphocytes; M: monocytes; N: neutrophils; E: eosinophils

All samples presented cellularity increase, with plasmatic cells prevalence.

 CONCLUSIONS: The meningeal myelomatosis, although rare, should be considered in patients with M.M. and their diagnosis is based on cytological studies and on the monoclonality demonstration of plasmatic cells present in CSF.

  

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PP09

SPINDLE CELL HEMANGIOMA WITH THE ATYPICAL LOCATION

 J. Talan-Hranilović1, M. Vučić1, D. Bedek2, K. Tomić1, T. Sajko3, V. Lupret3

 1Ljudevit Jurak University Department of Pathology; 2Department of Interventional Radiology and 3Department of Neurosurgery, Sestre milosrdnice University Hospital, Zagreb, Croatia

 
INTRODUCTION: Spindle cell hemangioma is a vascular tumor first described as a new entity in 1986 by Weiss and Enzinger, as a rare type of haemangioma. The tumor occurs usually in young adults and affects the subcutis of distal extremities or arose in the small bones of the hands, feet and tibia. Location within the spinal canal is extremely rare.

            CASE REPORT: A 31 year-old male patient complained on weakness of both legs that started in May 2005. In September 2005 he noticed paresthesias in both legs that slowly progressed toward his stomach. He was admitted at our Department of neurosurgery in March 2006 due to progressive weakness of his legs. Upon admission the neurological examination revealed a spastic paraparesis (3/5), hyperactive reflexes and paresthesias distal from Th10 dermatome. MR of the thoracic spine showed an intraspinal, extradural mass lesion, measuring 5,3 x 1,2 cm at the Th1-Th3 level.

Intraoperatively, C7-Th3 laminectomy was performed. On the right side of the spinal canal, a purple-grayish tumorous lesion was visualized. It was located extradurally compressing the dura to the left. The lesion was well demarcated from the dura, and it was removed completely in a microneurosurgical manner. Histologically H&E slides revealed a lesion which was composed of thin-walled cavernous vessels lined by flattened endothelial cells and containing thrombi. There were cellular spindled stromal areas between the cavernous spaces that were assumed to be representing collapsed small vascular canals. Some of endothelial cells contained prominent intracytoplasmatic vacuoles. Postoperative recovery was uneventful. Paresthesias regressed almost immediately after surgery and the amelioration of leg weakness was noticed.

            MATERIAL AND METHODS: Immunohistochemical analysis by CD 34, Vimentin, SMA and Ki-67 (DAKO, Copenhagen, Denmark) was performed in formalin fixed and paraffin embedded tumor tissues. As a visualization system labeled streptavidin biotin method (LSAB) was used on Dako Tech Mate automatic immunostainer using microwave streptavidin immunoperoxidase-MSIP protocol.

            DISCUSSION AND CONCLUSION: occasionally the spindle cell hemangioma is associated with Maffucci syndrome and was also seen in the Klippel-Trenaunay syndrome. The tumor arose in the vicinity of clearly abnormal vessels, supporting the idea that the spindle cell hemangioma is most likely a vascular malformation in which variation in blood flow gives rise to alternating areas of vascular expansion and collapse. Although, usually it was believed to be a tumor with limited metastatic potential and it is regarded as a benign tumor, about 60% of spindle cell hemangiomas recur and there is no evidence that these lesions have the ability to metastasize. This case is reported as a unique location of this rare type of hemangioma in the spinal canal.

 References:

1. WEISS SW, GOLDBLUM JR. Editors, Enzinger and Weiss’ s soft tissue tumors, (4th edition) Mosby, St.   Louis (2001) pp. 855-7.

2. KEEL SB, ROSENBERG AE. Hemorragic epitheloid and spindle cell hemangioma: a newly recognized, unique vascular tumor of bone. Cancer 1999; 85:1966-72. 

3. MENTZEL T, CALONJE E, FLETCHER CD. Vascular tumors of the skin and soft tissue. Overview of newly characterized entities and variants. Pathol 1994;15:259-70.

4. FLETCHER CD. Vascular tumors an update with emphasis on the diagnosis of angiosarcoma and borderline vascular neoplasms. Mongr Pathol 1996; 38: 181-206.

5. KEMPSON RL, FLETCHER CD, EVANS HL, HENDRICKSON MR, SIBLEY RK. Tumors of soft tissues. In: Rosai J and Sobin LH, Editors, Atlas of tumor pathology, fascicle 30, 3rd series, AFIP, WashingtonDC (2001), pp.307-70.

 

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PP10

PRIMARY OSTEOSARCOMA OF THE BREAST- A CASE REPORT

 L.Gashi-Luci1, Sh.Demaqi2, F.Kurshumliu1, A.Mustafa1

 1Institute of Anatomic Pathology, Prishtina, Kosovo; 2Clinic of Surgery, Thoracic surgery ward, Prishtina, Kosovo

 Osteosarcoma is a malignant bone producing tumor, very uncommon in extrasceletal sites.

This report describes a case of primary osteosarcoma of the breast arising in the lower medial quadrant of the right breast of a 50 years old female patient. The patient presented with a well demarcated node in close proximity to the areola. After preliminary examinations, the patient underwent radical mastectomy. On gross examination the tumor measuring 5.5 cm in diameter, well demarcated, firm, translucent, white-gray in color and with areas of necrosis was found. Histopathology revealed a tumor composed of a spindle to oval cell population with variable amounts of osteoid and areas of cartilage. The tumor cells were negative to PanCytokeratin. The diagnosis was consistent with osteosarcoma. Primary osteosarcoma is a rare mesenchymal tumor of the breast. This prompted us to report on a further case of this rare neoplasm.

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PP11

COMPARISON OF HER-2/neu BREAST CARCINOMA STATUS BETWEEN POŽEGA COUNTY AND ZAGREB

 I. Bagarić1, J. Fajdić,2 M. Vučić3, T. Leniček3, A. Dubravić3, T. Dolić4, M. Belicza3

 1Department of Pathology, 2Department of Surgery, General Hospital Požega; 3Ljudevit Jurak Department of Pathology, Sestre milosrdnice University Hospital; 4Roche d.o.o. Zagreb, Croatia

 
INTRODUCTION: HER-2/neu oncogen is located on chromosome 17q and encodes a transmembrane glycoprotein with intracellular tyrosine kinase activity. The HER-2/neu gene amplification or protein overexpression has been associated with a more aggressive breast tumor biology and the resistance to some types of chemotherapy. HER-2/neu overexpression is correlated with shorter disease-free and overall survival.

            AIM: The aim of the study was to evaluate and compare immunohistochemically determined HER-2/neu status of breast cancer patients in two Croatian regions (Požega and Zagreb). We analyzed data covering a 3- year period, between 2003 and 2005.

            PATIENTS AND METHODS: Immunohistochemistry for HER-2/neu (DAKO, Copenhagen, Denmark; donated by Roche d.o.o. Zagreb) was performed using labeled streptavidin biotin method (LSAB) visualization system on Dako Tech Mate automatic immunostainer with microwave streptavidin immunoperoxidasa-MSIP protocol. Semiquantification was performed using guidelines for scoring Hercep test by Dako.

            RESULTS: Immunohistochemistry was performed on 106 patients in Departments of the General Hospital Požega, and on 258 patients in Ljudevit Jurak Department of Pathology, University Hospital Sestre milosrdnice (Figure 1,2). The results of HER-2/neu testing in Požega County were: HER-2/neu score 0 in 61 patients; 1+ in 18 patients; 2+ in 15 patients and 3+ in 12 patients. Zagreb results were: HER-2/neu score 0 in164 patients; 1+ in 40 patients; 2+ in 15 patients and 3+ in 39 patients. The average age of HER-2/neu 3+ patients was 55.1 years in Požega County and 58.7 in Zagreb (Figure 3,4).

            CONCLUSION: HER-2/neu is a well established prognostic factor for breast cancer patients. Our results revealed that both groups of primary breast cancer (from Požega County and Zagreb) showed similar HER-2/neu distribution. There were no regional variations in HER-2/neu status distribution in breast cancer patients of two Croatian regions. Evaluation of HER-2/neu status in combination with other prognostic parameters should help in determination of breast cancer prognosis and therapy. HER-2 over-expression provides a new target in breast cancer therapy with development of monoclonal antibody targeted against HER-2 – trastuzumab (Herceptin®).

References:

1. LIPTON A et al. Cancer 2005;104:257-63.
2. JAKIĆ- RAZUMOVIĆ J et al. Cancer Therapy 2005; 3:167-76.
3. VUČIĆ M et al. Acta Clin Croat 2003;42:183-8.

 

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PP12

MALIGNANT PHYLLODES TUMOR WITH THREE DIFFERENT SARCOMATOUS COMPONENTS – A CASE REPORT

Tatjana Bujas1, Mira Mlinac Lucijanić1, L. Tomić2, P. Kirac3, D.Tomas3

1Department of Pathology, General Hospital Karlovac, Croatia; 2Department of Gynecology and Obstetrics, Josip Benčević General Hospital; Slavonski Brod, Croatia; 3Department of Surgery and Ljudevit Jurak University Department of Pathology, Sestre milosrdnice University Hospital, Zagreb, Croatia

 
INTRODUCTION: Malignant phyllodes tumor of the breast is a rare biphasic neoplasm. Its stroma shows frankly sarcomatous, usually fibrosarcomatous changes. Heterologous differentiation such as liposarcoma, osteosarcoma, chondrosarcoma or rabdomyosarcoma may occur.

CASE REPORT: A 71year-old woman was admitted because of a painless lump in her right breast. Physical examination revealed a movable mass, about 3 cm in diameter, in the upper outer quadrant of the right breast. The patient underwent ultrasonography and mammography showing a 3.5 cm oval and relatively well circumscribed mass with coarse, amorphous calcifications (Fig. 1). Right breast 20x16x4.5 cm and axilla 9x7x3 cm were admitted for histopathological examination. In the upper outer quadrant a whitish-gray, hard mass with infiltrative borders that measured 3.3 cm in maximum diameter was found. Microscopically, the tumor was consisted of three different sarcomatous components. The most prevalent was made of osteoid surrounded by proliferating atypical osteoblasts and multinucleated, osteoclast-like giant cells. Focally, trabeculae of woven bone with coarse calcifications surrounded with atypical osteoblasts were also noted (Fig. 2). Second sarcomatous component contained lipoblasts, atypical mesenchymal cells with high mitotic activity, and mature lipomatous cells (Fig. 3). In some areas the tumor was composed of round to oval atypical cells with scanty, poorly outlined cytoplasm in the chondromatous matrix with partially myxoid changes (Fig. 4). The entire tumor was sampled for histology to find its epithelial component. In few additional samples epithelial component was found which consisted of luminal epithelial and myoepithelial cells surrounded with hypercellular, mitotically active stroma that showed enhanced intracanalicular growth pattern with leaf-like projections into dilated lumens (Fig 5). Immunohistochemical analysis (CK, EMA, SMA, desmin, S-100) confirmed existence of three different sarcomatous components. Nine lymph nodes measuring from 1 to 5 cm were found in axillary fat tissue and were without signs of the tumor. The diagnosis of malignant phyllodes tumor with associated osteosarcomatous, chondrosarcomatous and liposarcomatous differentiation was established.

            DISSCUSION: To our knowledge this is the second report in English literature of malignant phyllodes tumor with associated different sarcomatous components. The first case of a 96-year-old female patient with malignant phyllodes tumor, which stroma contained elements of leiomyosarcoma, liposarcoma, rhabdomyosarcoma and malignant fibrous histiocytoma, was reported by Guerrero et al. Due to overgrowth of the sarcomatous components malignant phyllodes tumor may be confused with pure sarcomas of the breast or with carcinosarcomas. In our case, because of three-associated sarcomatous components the main differential diagnosis was carcinosarcoma. After surgical excision, careful gross examination and thorough sampling of the specimen is recommended to establish definitive diagnosis. In our case, evidence of malignant epithelial component was not found and the diagnosis of malignant phyllodes tumor with associated osteosarcomatous, chondrosarcomatous and liposarcomatous stromal differentiation was established.

 References

1. TAVASSOLI FA, DEVILEE P. (Eds.) World health organization classification of tumours. Pathology and genetics of tumours of the breast and female genital organs. Lyon: IARC Press, 2003, pp 9-110.
2. SILVER SA, TAVASSOLI FA. Osteosarcomatous differentiation in phyllodes tumors. Am J Surg Pathol 1999; 23: 815-821.
3. VERA-SEMPERE F, GARCIA MARTINEZ A. Malignant phyllodes tumor of the breast with predominant chondrosarcomatous differentiation. Pathol Res Pract 2003; 199: 841-845.
4. GUERRERO MA, BALLARD BR, GRAU AM. Malignant phyllodes tumor of the breast: rewiev of the literature and case report of stromal overgrowth. Surg Oncol 2003; 12: 27-37.

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 PP13

METASTATIC AND SURVIVAL RATE IN PATIENTS WITH ORAL CARCINOMA ACCORDING TO THE TUMOR’S MORPHOLOGICAL FEATURES

 V. Janevska1, L. Spasevska1, G. Petrusevska1, S. Kostadinova1, D. Popovic2, R. Jovanovic1

 1Institute of Pathology; 2Clinic of Maxillofacial Surgery, Skopje, Macedonia

 AIM OF THE STUDY: is to analyze morphological features of oral spinocellular carcinomas in patients treated with surgical therapy only and to find out which of them influence the metastatic and survival rate.

MATERIAL AND METHODS: 75 cases of oral carcinomas were analyzed using: 1. clinical parameters 2. radiological methods (US, KT, MRI) 3. histopathological parameters 4. type of surgical intervention. Analyzed histopathological parameters were: degree of cellular differentiation, nuclear pleomorphism, number of mitoses, type of tumor invasion, inflammatory reaction of the host tissue, desmoplasia, invasion into vascular channels and the depth of tumor’s invasion. The malignancy was graded according to the Nason Richards system.

The follow up period was 4 to 6 years.

RESULTS: Sixty patients had simultaneous neck dissection and in 15 cases the lymph nodes were followed ¸up through regular monthly control. The depth and the type of tumor’s and vascular invasion were strongly associated with occurrence of neck metastases. The survival period was connected to the degree of differentiation, vascular invasion, presence of neck metastases and the tumor size (T).

CONCLUSION: There are strong prognostic histological factors in oral carcinomas and simultaneous neck dissection is recommended in these patients.

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PP14

OCCULT PAPILLARY CARCINOMA OF THE THYROID GLAND PRESENTING AS CERVICAL CYSTIC MASS – A CASE REPORT

 K Horvat1, A Borovečki1, A Škrtić1, S Džebro1, T. Znaor2, S. Gašparov1, M. Dominis1

 1Department of clinical pathology and cytology, 2Department of otorhinolaryngology, University Hospital Merkur, Zagreb, Croatia

 Occult papillary carcinoma of the thyroid has been defined as a papillary carcinoma measuring 1 cm or less in diameter. A solitary cystic mass in the lateral aspect of the neck may rarely be the only initial presenting symptom of occult papillary carcinoma.

A 50 year old male patient presented with a palpable mass localized in the anterolateral neck region, in the middle third of m. sternocleidomastoideus. The mass was evaluated by ultrasound, which revealed an inhomogenous cystic mass. A FNA of the mass was suspect to thyroid neoplasm. A technetium-99 thyroid scan suggested only normal uptake. Ultrasound of the thyroid gland was not performed.

Resected mass consisted of fat tissue with an encapsulated oval cystic tumor, 3x2x1,5 cm and six lymph nodes, 0,4-1 cm in diameter. The oval cystic mass morphologically revealed papillary carcinoma, CK7, TTF-1, thyroglobulin positive, CK20 negative. Remaining six lymph nodes showed only reactive features and fat tissue without metastases. The dilemma was either of ectopic thyroid tissue transforming in papillary carcinoma in branchial cleft cyst or the lymph node metastasis from a primary thyroid tumor. Complete examination of thyroid gland was suggested.

Total thyroidectomy and modified neck dissection was performed. There were no macroscopic changes on thyroid gland. Histological examination revealed two foci of papillary carcinoma, measuring 0,6 and 0,3 cm in diameter. The neck dissection specimen exhibited 12 lymph nodes without metastatic papillary carcinoma.

 References

1. SEVEN H et al., Incidence of occult thyroid carcinoma metastases in lateral cervical cyst. Am J Otolaryngol 2004;25:11-17.
2. MONCHIK JM et al., Occult papillary carcinoma of the thyroid presenting as a cervical cyst. Surgery 2001;129:429-32.
3. CHAN JKC, Thyroid and parathyroid in: Weidner N et al. Modern surgical pathology. Philadelphia: Elsevier Science, 2003:1714-5.

 

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PP15

MICROVESSEL DENSITY AND MAST CELL DENSITY CORRELATE WITH IMMUNOHISTOCHEMICAL EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR IN GASTRIC CARCINOMA

 Ovidiu Mederle1, Sorin Ioanovici2, Marius Raica1, Anca Maria Cimpean1, Dragos Izvernariu1

 1Department of Histology, Victor Babes University of Medicine and Pharmacy; 2Department of Gastroenterology, Victor Babes University of Medicine and Pharmacy, Timisoara, Romania

 AIM OF THE STUDY: is to investigate the relationship between VEGF, mast cells, and microvessel density on gastric carcinoma.

MATERIALS AND METHODS: 32 cases with gastric cancer (T1–2 cases, T2–11 cases, T3–19 cases) were investigated. Specimens were processed according to the conventional histological procedure, and slides were stained with haematoxylin-eosin. Additional slides were immunostained for polyclonal VEGF and combined CD34 – alcian blue- safranin pH0.2. VGEF reaction was scored as negative (0), weakly positive (+), moderate (++), and intense (+++). The mast cells and blood vessels were counted on the same slides, using the hot spot method. Images were captured as JPEG, and analyzed with Lucia G version 6.1

RESULTS: Adenocarcinoma of intestinal type was found in 18 cases, and the diffuse type was noticed in 14 cases. VEGF was expressed in 22 out of 32 patients (15 with strong cytoplasmic reaction with granular pattern). The expression of VGEF did not correlate with pathologic type or stage of the tumor. On the other hand, VEGF was weakly or moderately positive in intestinal metaplasia and dysplasia found in the carcinoma periphery, however, it was constantly negative in the normal mucosa. We found an increased number of microvessel and mast cells in 19 out of 22 VEGF positive cases. In VEGF positive cases the median microvessel density (MVD) was 47.2, and mast cell density was 38.5 (x200). In VEGF negative cases, MVD was 29.2, and the mast cell density was 24.7.

CONCLUSIONS: Both MVD and mast cell density correlate with immunohistochemical expression of VEGF. Our results suggest that mast cells are involved in gastric cancer associated angiogenesis, and that such tumors are candidates for targeted therapy with VEGF inhibitors.

 

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PP16

GASTROINTESTINAL MELANOMA – REVIEW OF THE COMPUTER DATA BASE IN THE PERIOD 1996-2005.

T. Bujas1, K. Tomić2, M. Perić Balja3, S. Domitrović-Krušlin4, D. Baličević4, B. Krušlin4

 1Department of Pathology, General Hospital Karlovac, Croatia; 2Department of Pathology, General Hospital Dr. Josip Benčević, Slavonski Brod , Croatia; 3Department of Pathology, University Hospital for Tumors, Zagreb, Croatia; 4Ljudevit Jurak Department of Pathology, Sestre milosrdnice University Hospital, Zagreb, Croatia

 AIM OF THE STUDY: Malignant melanoma is very rare in the gastrointestinal system; it comprises about 0.2-3% of all melanomas and has extremely poor prognosis. Surgical treatment ranges from radical resection with lymphadenectomy to local excision alone. Primary melanoma is most frequently found in anorectal region of the gastrointestinal system.

Aim of the study was to analyze frequency and sites of primary gastrointestinal melanoma in our bioptic material collected during a 10 year period.

MATERIALS AND METHODS: We used the computer database from Ljudevit Jurak University Department of Pathology and Surgical Department, Sestre milosrdnice University Hospital, Zagreb in the time period from 1996 to 2005.

RESULTS: During the analyzed period there were 2831 patients (M:F=1748:1083) with diagnosed gastrointestinal tumors. Female patients were between 22-86 years old (mean 62,0) while males were between 26-84 (mean 68,0). In the examined time period malignant melanoma was diagnosed only in 16 (0,6 %) patients (M:F=12:4).

At the time of diagnosis male patients with melanoma were between 22-79 years old (mean 54,0), while females were between 42–70 (mean 59,0). Out of all patients with gastrointestinal melanomas only 5 (31.3%) cases (M:F=3:2) were primary gastrointestinal, while 11 (68.7%) cases were metastases from skin melanoma. Two cases of primary melanomas in gastrointestinal system were originated from gallbladder (M:F=1:1), while 3 cases were diagnosed in the anorectal region (M:F=3:1). Female patients with primary gastrointestinal melanoma were 52 and 57 years old, while males were between 52 and 75 (mean 66,0). Metastastatic melanomas usually involved small intestine (9/11 cases), while only one case was found in stomach and one case in colon.

CONCLUSION: Although primary gastrointestinal melanomas are very rare, they are almost never suspected on clinical examination. In our bioptic material primary melanomas were found in anorectal region and gallbladder, while metastatic melanomas were usually found in the small intestine.

 References

1. PANTALONE D. et al. Eur J Surg 2001;166:583-4.
2. KIM C.J. et al. Cancer Control 2002;9:9-15.
3. POGGI SH. et al. J Clin Gastroenterol 2000;30:441-4.
4. MÜLLER D. et al. Acta Clin Croat 2001;40:203-7.
5. VUČIĆ M. et al. Acta Clin Croat 2001;40:287-91.

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 PP17

TUMOR BUDDING AS A PARAMETER OF AGGRESSIVENESS IN COLORECTAL CARCINOMA

 L. Spasevska, V. Janevska, V. Janevski, R. Jovanovic, S. Kostadinova-Kunovska, V. Filipovski

 Institute of Pathology, Faculty of Medicine, Skopje, Macedonia

 AIM OF THE STUDY: To evaluate the significance of tumor budding in correlation with histopathological parameters and distant metastasis.

METHODS: Surgically resected specimens from 39 pT2 or pT3 colorectal adenocarcinomas were studied. The presence of tumor budding was examined according to Moradomi`s criteria using hematoxylin-eozin stained sections. Budding was divided into two grades based on the number of “budding” foci within a microscopic field of x250: high-grade budding (≥10 foci in a field) and low-grade budding (<10 foci in a field).

RESULTS: Budding was observed in 79,2% of the lesions, 41,2% of them with high-grade, and showed strong correlation with the histopathological characteristics which are generally thought to represent tumor aggressiveness, such as extramural spread, infiltrative growth pattern, number of lymph nodes involved, extramural venous invasion and distant metastasis (p<0,05). Significant correlation between the tumor budding and the tumor differentiation, tumor type, lymphocyte infiltration, and maximum diameter of the tumor, were not found. Multiple regression analysis for all parameters analyzed, showed the strongest impact of tumor budding on the number of lymph nodes affected (b=0,69, p=0,015; correlation coefficient 0,69, p<0,05).

CONCLUSION: Tumor budding would be a simple and inexpensive marker for estimating aggressiveness of colorectal adenocarcinoma, as well as, for predicting lymph node metastases.

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 PP18

HISTOLOGICAL TYPES OF LYMPHOMA IN THE GASTROINTESTINAL SYSTEM DURING 10 YEARS IN OUR BIOPTIC MATERIAL

 T. Bujas1, K. Tomić2, M. Perić Balja3, H. Čupić4, D. Baličević4, M. Belicza4

 1Department of Pathology, General Hospital Karlovac, Croatia; 2Department of Pathology, General Hospital Dr. Josip Benčević, Slavonski Brod, Croatia; 3Department of Pathology, University Hospital for Tumors, Zagreb, Croatia; 4Ljudevit Jurak Department of Pathology, Sestre milosrdnice University Hospital, Zagreb,Croatia

 AIM OF THE STUDY: Gastrointestinal tract is the most commonly involved extranodal site for non-Hodgkin lymphoma that usually affects the stomach, less frequently small intestine and very rarely colon or esophagus.

Aim of the study was to analyze frequency and types of gastrointestinal lymphoma in our bioptic material during the last 10 years.

MATERIAL AND METHODS: We used the computer database (Thanatos) from Ljudevit Jurak University Department of Pathology for the time period from 1996-2005 for all patients who underwent gastrointestinal tract surgical procedures at the Department of Surgery, Sestre milosrdnice University Hospital, Zagreb.

RESULTS: In the examined time period there were 2828 patients (M:F=1750:1078) presenting with gastrointestinal tumors. Esophagus was involved in 108 cases, stomach in 720 cases, and 2000 were in the small intestine and colon.

Gastrointestinal lymphoma was diagnosed in only 18 (0,6 %) cases (M:F=9:9). Male patients were between 50 and 73 years of age (mean 60,0 years); while females were between 49-82 (mean 65,0). The majority (16/18) of gastrointestinal tract lymphomas were found in the stomach (M:F=7:9), while only two cases were diagnosed in the small intestine. Both patients with small intestine lymphoma were male, 64 and 67 years old. Histologically all examined lymphomas were classified according to the World Health Organization (WHO) as diffuse large cell lymphoma, B phenotype.

CONCLUSION: Gastrointestinal lymphoma was more commonly diagnosed in the sixth decade of life, with stomach being the predominant site of involvement.

In patients having MALT lymphoma diagnosed on endoscopic biopsy, surgical operation was not performed. Therefore confirmation of MALT lymphomas was impossible.

Lymphomas involving the gastrointestinal tract, although rare, should be considered in differential diagnosis when dealing with gastrointestinal tract malignancies.

 References

1.       KONIARIS LG. et al. J Am Coll Surg 2003;197:127-31.

2.       GUERNEY KA. et al. Br J Cancer 1999;79:1922-34.

3.       LONGO WE. et al. Am Surg 1995;61:495-500.

4.       KURODA T. et al. J Gastroenterol 1996;31:437-40.

 

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PP19

CORTICOSTEROIDS AND NOT BETA – CAROTENE POTENTIATES GASTRIC CARCINOMA IN THE RAT EXPERIMENTAL MODEL INDUCED BY DMBA

 Narcisa Radbea1, Ovidiu Mederle2, Gheorghe Darabus1, Horatiu Papiu3, Marius Raica2

 1Faculty of Veterinary Medicine, Timisoara, Romania; 2Victor Babes University of Medicine and Pharmacy, Department of Histology, Timisoara, Romania; 3County Hospital, Department of Surgery, Arad, Romania

 AIM OF THE STUDY: is to create an experimental model of gastric carcinoma in the rat using dimethylbenzantracene (DMBA), and to evaluate the effect of corticosteroids and beta-carotene.

MATERIAL AND METHODS: The experiment was performed with Sprague-Dowley adult rats (250-300 g).The control group included five animals which did not receive DMBA. DMBA was administered in tap water (concentration 0.01 mg/ml) for all other animals (n=25). Two subgroups (2 and 3) received only DMBA, and were sacrificed after 6 and 12 months. Other two subgroups (4 and 5) received DMBA and dexamethasone, and the last subgroup received DMBA, dexamethasone and beta-carotene. The stomach was removed and paraffin blocks were performed from all anatomic areas.

RESULTS: We did not find any dysplastic, metaplastic or malignant lesion in the control group. Only dysplasia in one case, and metaplasia of colic type in two cases were found in the second group. In the third group, treated with DMBA only for 12 months, we found a gastric carcinoma (diffuse type) in one case, and severe dysplasia in two cases. In the 4th subgroup 2 cases had gastric carcinoma (diffuse type), and two cases severe dysplasia. In the 5th subgroup we found two gastric carcinomas and one dysplasia. The microscopic examination showed only diffuse type gastric carcinoma, severe dysplasia, and type II of intestinal metaplasia, as demonstrated by histochemistry. The proliferation rate assessed with PCNA and Ki67 was similar in gastric carcinoma and severe dysplasia.

CONCLUSION: The experimental model of gastric cancer induced with DMBA in the rat showed the development of a malignant lesion in 5 out of 20. Dysplasia was found in 6 cases. Gastric carcinogenesis seems to be stimulated by the presence of dexamethasone, and was not significantly influenced by beta-carotene.

 

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PP20

ORTHOTOPIC LIVER TRANSPLANTATION DUE TO MULTIPLE METASTASES OF SOLID-PSEUDOPAPILLARY TUMOR OF THE PANCREAS – CASE REPORT

 M. Milković Periša, A. Škrtić, A. Borovečki, S. Gašparov, S. Džebro, M. Dominis

 Department of Clinical Pathology and Cytology University Hospital Merkur, Zagreb, Croatia

 A 20 year-old woman was admitted to our hospital three years ago because of a pain in the upper left abdomen. Ultrasound showed a round, solitary, encapsulated tumor of the pancreas. A partial pancreatectomy and spleenectomy were done. The encapsulated tumor was localized in the pancreas and measured 10 cm in greatest diameter. The tumor was necrotic, pseudocystic and partially papillary. Morphological features consisted from solid and pseudopapillary structures with eosinophil monomorfic cells with small, round to oval nuclei. A solid-pseudopapillary tumor of the pancreas is usually a benign neoplasm affecting predominantly young women. Despite its benign biological behavior solid-pseudopapillary tumor can produce metastases in regional lymph nodes, liver, peritoneum and greater omentum. She was well untill January 2006 when she was presented with multiple liver tumors found by ultrasound and CT. FNA revealed tumor cells of the same type as in the primary tumor. The only effective therapy in such cases is enucleation of metastatic tumors. Due to metastases in different segments of the liver the enucleation of tumors was not possible. Split living donor transplantation was performed in March 2006. There were 16 metastatic nodules found in the liver ranging in size between 0.5-2 cm. The patient is alive and well.

 References

1. KLÖPPEL G et al. Solid-pseudopapillary neoplasm in: Hamilton SR, Aaltonen LA, eds. Pathology&Genetics: Tumors of the Digestive System. Lyon: IARCPress, 2000:246-248.
2. SAIURA A et al. Hepatogastroenterology 2000;47:887.

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 PP21

MYXOID LEIOMYOSARCOMA OF THE LIVER

 M. Sičaja1, D. Romić1, D. Rahelić2, Asraa Namiq3, J. Forster3, I. Damjanov3

 1Medical students, The University of Zagreb School of Medicine, Zagreb, Croatia, 2Clinical Hospital Dubrava, Zagreb, Croatia, 3The Department of Pathology, The University of Kansas School of Medicine, Kansas City, Kansas, USA

 Myxoid leiomyosarcoma is a subtype of malignant smooth muscle cell tumors originally recognized for the first time in the uterus. This is a report of a myxoid leiomyosarcoma arising in a cirrhotic liver. The tumor was resected from a 64-year-old man. On gross examination, it was soft and hemorrhagic. The mass did not have a distinct capsule, but was nevertheless sharply demarcated from the surrounding liver parenchyma. The tumor was composed of deceptively benign looking smooth muscle cells with clear cytoplasm suspended in a myxoid stroma with foci of hemorrhage. Routinely performed immunohistochemical stains showed that the tumor cells were positive for vimentin and smooth muscle actin. Tumor cells did not react with the antibodies to desmin, keratins, S100, CD31, CD34, CD117, coagulation factor VIII, and melanocytic antigens (Melan-1 and HMB45). The immunohistochemistry for MIB-1 disclosed a moderate proliferative activity. Immunohistochemistry and electron microscopy confirmed that this was a smooth muscle cell neoplasm. The abundance of glycogen and ultrastructural signs of smooth muscle differentiation were considered consistent with an immature smooth muscle cell phenotype consistent with the diagnosis of myxoid leiomyosarcoma. In contrast to the uterine tumors, the cells of the present tumor contained abundant cytoplasmic glycogen and thus resembled more fetal than adult smooth muscle cells. We suggest that this ‘‘fetal phenotype’’ may account in part for the propensity of myxoid leiomyosarcomas to grow more aggressively and to metastasize more often than other low-mitotic smooth muscle cell tumors. Since myxoid leiomyosarcomas are aggressive tumors, it is important to recognize them histologically and also bear in mind that these tumors can occur even in unusual extrauterine locations such as a cirrhotic liver.

 

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PP22

FINE NEEDLE ASPIRATION CYTOLOGY OF PULMONARY HAMARTOMAS.

CYTOLOGYCAL STUDY OF 11 CASES

 E. Garcia-Ureta, E. Carro Rey, O. Robles Viega, R. Alvarez Rodriguez

 Citologia Servicio De Anatomía Patológica Hospital Universitario Juan Canalejo, La Coruña, Spain

             INTRODUCTION: The lung hamartoma, although rare, is a benign tumor more frequent in lungs; its approximate incidence is 0,25 % in the general population. Cytological studies of these processes are very reduced, partly due to the difficulty of obtaining material of cartilaginous lesions and partly for the difficulty of interpreting these findings. Present work wants to expose the results of 11 cases diagnosed by cytology.

            MATERIAL AND METHODS: We have assessed cytological findings of 11 cases (8 men and 3 women), picked up in our service during the years 1990 until 2005, with a cytological diagnosis of lung hamartoma in the material obtained by PAAF and confirmed histologically. The needle biopsy technique and the handling of the aspired material were carried out and routinely processed (MGG. Pap).

            RESULTS: The cellularity was moderate to abundant in 6 cases and scarce in 5 cases. Combined presence of epithelial and mesenchymal components have been confirmed in 7 cases. The existence of epithelial cells was observed in 7 cases, generally small, uniform in size and forms, without mitosis or atypia presence. The fibromyxoid tissue was present in 10 cases, varying from lax and delicate to more cohesive and structured. The chondroid material varied from myxocartilage to mature cartilage and was confirmed in 7 cases, the presence of well-formed cartilage was found in 4 cases, which helped greatly in establishing the diagnosis since its sole presence can be explained in very few situations. In 3 cases adipose tissue was shown, sometimes in continuity with fibromyxoide or condroide tissue. The existence of smooth muscle fibers was demonstrated in 2 cases. In 4 cases a discreet inflammatory cellularity was confirmed, formed by some few lymphocytes and histiocytes, necrotic material not being evident. These cytological features were a reflection of the lung hamartomas composition.

            CONCLUSIONS: These cytologyic findings confirm the value of the PAAF in the diagnosis of the lung hamartoma. The best results for assessment of cytological FNAC findings in lung hamartomas can be achieved by interpretation of the cytological findings within the context of clinical and radiological findings, so they should be used previous to any aggressive diagnosis procedure, especially in view of the low risk of complications.

 

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PP23

SEVERE PULMONARY HYPERTENSION IN INFANTS WITH CONGENITAL HEART MALFORMATIONS

 S. Duganovska, G. Petrusevska, B. Bogoeva, S. Kostadinova, V. Janevska, S. Banev, R. Jovanovic

 Institute of Pathology, Medical Faculty, Skopje, Macedonia

 We present a study of 4795 autopsies performed during a period of 8 years at the Institute of Pathology in Skopje. We analysed the morphology of congenital heart malformations (CHM) and changes in lung blood vessels in cases of developed left-right shunt. We used standard tissue staining: HE, Van Gieson-Elastica, trichrom Masson and Reticulin. We performed histomorphometrical analysis using the LUCIA M – NIKON Image analysing system. The thickness of the media of the lung muscle arteries has been measured and then compared to the complete total diameter of the blood vessel. Thickness coefficient obtained was compared to the graduation results according to the Heath-Edwards classification (I, II, III, IV) and to the control group. The changes in lung blood vessels in a group of 38 cases of CHM were classified according to the above mentioned classification system. Most of the cases were classified in group I (63,1%), group II (18,4%) regarding the cases with VSD, AV-canal, PDA, ASD/VSD; group III (7,8%) regarding the cases with VSD, CoA, TGA+VSD; group IV (10,5%) regarding the cases with VSD, AV-canal, Single ventricle. Microscopically, there were aneurismal, plexiform and dilatation lesions of the blood vessels. Results obtained from this study emphasize the necessity of lung biopsy while evaluating reversible and irreversible phases of the disease, which is very important for further surgical treatment of children with CHM.

 

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PP24

CYTOLOGICAL FINDINGS IN A CASE OF PRIMARY PULMONARY PLASMACYTOMA (PPP)

 E. Garcia-Ureta, E. Carro Rey, O. Robles Viega, R. Alvarez Rodriguez

 Citologia Servicio De Anatomía Patológica Hospital Universitario Juan Canalejo, La Coruña, Spain

             OBJETIVE: Extramedullary plasmacytoma is a rare immunoproliferative monoclonal disease and only a few cases diagnosed by cytological studies have been reported. It has to be differentiated from other neoplasm and reactive plasma cell proliferations. Cytological findings of bronchial brushing and washing of PPP are shown in the case report.

            CASE REPORT: A 58 year-old man was hospitalized for assessment of a right hilar mass that was noted on a chest radiograph; laboratory data were all within normal limits. Material for cytological and histological examination was obtained during fiberoptic bronchoscopy from the endobronchial lesion. Brushing and washing cytological samples were processed by conventional preparations, Thin-Prep technique, staining techniques and immunocytochemistry.

            RESULTS OF CYTOLOGICAL FINDINGS: Cytological examination reveals a dispersed population of cells with a plasmacytoid appearance with eccentric nuclei, abundant cytoplasm and the characteristic paranuclear hof. Binucleation was common and mitotic figures were easily identified. Inmunocytochemical stains revealed that the tumor cells stained for kappa light chain, the cytological diagnosis was plasmacytoma and was confirmed histologically. Work up to rule out multiple myeloma was completed. Skeletal survey showed no other sites of involvement. Serum electrophoresis was normal and Bence Jones proteinuria was absent. Bone marrow examination demonstrated no abnormalities. Based on these findings an extramedulary plasmacytoma of the lung (PPP) was diagnosed.

            CONCLUSIONS: Cytology material of brushing and washing the affected area can be a safe and fast procedure for diagnosing myeloma of the lung. Mainly, these tumors need to be distinguished from reactive inflammatory processes, plasmacytoid lymphoma and plasma cell granuloma.

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PP25

THYMOMA ASSOCIATED WITH MULTIPLE PRIMARY NEOPLASMS IN A SINGLE PATIENT

 S. Šitić1, L. Brčić2, Z. Juroš3, I. Nikolić3, B. Krušlin4

 1Clinical Hospital Merkur, Zagreb, Croatia; 2University of medicine, Zagreb, Croatia; 3Clinic for pulmonary diseases, Zagreb, Croatia; 4Ljudevit Jurak University Department of Pathology, Sestre Milosrdnice University Hospital, Zagreb, Croatia

 We present a case of a 66 year old woman with three primary neoplasms. Her past medical history was notable for mild hypertension, old CVI and she has been under psychiatric supervision due to personality disorder for a year.

In 2003, she underwent surgery due to breast cancer and was treated with chemotherapy. In the same year, after radiological investigation, a well circumscribed mass of an unknown origin was detected in the apical region of the left lung. Extensive clinical investigation followed and colorectal adenocarcinoma was diagnosed. She underwent surgical resection. She was treated with another chemotherapy for colorectal adenocarcinoma but nevertheless, previously detected mass in the lung seemed to progress. During the preoperative analysis another tumor mass was detected in the posterior mediastinum. Both, mediastinal and lung tumors were resected, consequently, lung adenocarcinoma and mediastinal thymoma were diagnosed.

According to the literature, it is well known that thymoma is associated with an increased risk of other malignancies. The etiopathogenetic mechanism is still unknown.

It seems that extensive clinical examination and long-term follow-up of patients with thymoma should be recommended.

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PP26

IPSILATERAL RENAL CELL CARCINOMA AND ADRENAL PHEOCHROMOCYTOMA

 S. Shala1, I. Brigić2, K. Tomić3, R. Mati1, V. Kotori1, A. Mustafa1, D. Tomas4 (Kosovo, Croatia)

 1Institute of Anatomic Pathology, Prishtina, Kosovo; 2Department of Urology Sestre milosrdnice University Hospital, Zagreb, Croatia; 3Department of Pathology, General Hospital Slavonski Brod, Croatia; 4Ljudevit JurakUniversity Department of Pathology, Sestre milosrdnice University Hospital, Zagreb, Croatia

 This report describes a case of a 66 year-old male presenting with a well-demarcated yellowish tumor mass with yellowish color, measuring 3.5 cm near the hilum of the right kidney and centrally located well-demarcated reddish mass of the right suprarenal gland measuring 2 cm in diameter.

Histopathology of the renal mass revealed a tumor composed of atypical epithelial cells with abundant clear cytoplasm and nucleus with prominent nucleoli arranged in alveolar and solid growth pattern consistent with Renal Cell Carcinoma. While mass in suprarenal gland was composed of polymorphic and polygonal tumor cells with abundant granular cytoplasm arranged in clusters consistent with Pheochromocytoma.

The association of Renal Cell Carcinoma and Pheochromocytoma is infrequent and mostly found in constitution of von Hippel-Lindau disease.

 

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PP27

TUMORS OF THE ADRENAL CORTEX

 B. Bogoeva, S. Banev, B. Krstevska, V. Filipovski, S. Kostadinova

 Institute of Pathology, Clinic of Endocrinology, Skopje, Macedonia

             INTRODUCTION: Integrated research by clinical endocrinologists, urologists and histopathologists had resulted in major advances in understanding of the adrenal and its diseases, especially differentiation between malignant and benign tumors.

            THE AIM OF THE STUDY: to report histopathological findings of adrenocortical tumors.

            MATERIAL AND METODS: We analyzed 22 cases with adenoma and 12 with carcinoma (2 cases with metastases) which occurred in the period of last 10 years.

Clinical manifestation: abdominal mass and pain, hormonal excess, hypertension, Cushing’s syndrome with or without virilization, primary hiperaldosteronizam etc.

Histopathological examinations were performed by light microscopy after routine staining: HE, PAS and Sudan and immunohistochemical: Ki 67, chromograninA, NSE.

The differentiation between benign and malignant tumors was done on a summation of different parameters:tumor weight (benign < 100g and malignant >150g), encapsulation, preservation of mainly normal or mainly abnormal structure, nuclear atypia (slight, moderate or strong atypia), nuclear hiperchromasia (moderate or marked), structure of nucleoli (normal or abnormal), mitotic activity defined as number of mitotic figures per ten high-power fields (two, more than two), regressive changes such as necrosis, hemorrhage, fibrosis or calcification (little or no change, moderate or extensive), invasion of the capsule and\or of the blood vessel (no invasion, invasion).

            CONCLUSION: Differentiation between malignant and benign tumors is difficult. The highest discriminating parameters were invasion of the capsule or the blood vessel wall, followed by nuclear hiperchromasia, mitotic activity and regressive changes. They corresponded with high tumor weight.

 

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PP28

ADRENAL GLAND SCHWANNOMA MIMICKING BREAST CANCER METASTASIS – A CASE REPORT

 M. Perić Balja1, K. Tomić2, T. Bujas3, M. Ulamec4, A. Reljić5, B. Krušlin4

 1Department of Pathology, University Hospital for Tumors, Zagreb, Croatia; 2Department of Pathology, General Hospital Dr. Josip Benčević, Slavonski Brod, Croatia; 3Department of Pathology, General Hospital Karlovac, Karlovac, Croatia; 4 Ljudevit Jurak University Department of Pathology, Sestre Milosrdnice University Hospital, Zagreb; 5Department of Urology, Sestre Milosrdnice University Hospital, Zagreb, Croatia

             INTODUCTION: Schwannoma is a benign, usually encapsulated nerve sheath tumor originating from Schwann cells. Typically, schwannomas affect the cutaneous nerves of the head and neck, upper and lower extremities and trunk. They predominantly occur in females between the second and fifth decade of life. Visceral schwannomas are very rare, and according to literature were described in the heart, kidney and lung. The retroperitoneal space is another localization of schwannomas. Except in cases of von Reclinghausen´s disease, the adrenal localization of schwannoma is particularly rare.

            CASE REPORT: We present a case of a 55 year-old female patient with previously diagnosed breast cancer treated with mastectomy followed by lymphadenectomy. Histologically, breast cancer was classified as ductal invasive breast cancer, histological grade 2, without lymph nodes metastases. Hormone receptors (estrogen and progesterone) were highly positive while HER2/neu was negative. Clinical examination did not find any signs of neurofibromatosis.

Patient was treated with hormonal therapy. Three months after mastectomy ultrasound examination revealed a hormonally inactive tumor in the left adrenal gland that was suspected for a metastatic process. After adrenalectomy, macroscopic examination revealed a partially encapsulated tumor within the adrenal gland with yellowish solid cut surface, measuring 5x4x3 cm.

Microscopically, the tumor was composed of solid sheets and bundles of uniform spindle shaped cells, along with various vascular and inflammatory elements embedded in an extensive extracellular matrix. No Verocay bodies were found. Immunohistochemicaly, tumor cells were positive for S-100 protein. Extensive histological analysis revealed no ganglion cells.

            DISCUSSION: Histological and immunohistochemical analysis of described adrenal gland tumor was consistent with the diagnosis of schwannoma. Adrenal tumors are frequently incidental and asymptomatic discoveries, and their therapy is a subject of controversial discussion. The combination of adrenal gland schwannoma and primary breast cancer is very rare, very probably incidental, but preoperatively difficult to distinguish from metastasis.

 References:

1. ARENA V et al. Folia Neuropatol 2004; 42:177-9.

2. TERADA T et al. Virchows Arch 2004; 444:95-97.

3. SCHEITHAUER BW, WOODRUFF JM, ERLANDSON RA. Tumors of the Peripheral Nervous  System, Armed Forces Institute of Patology, 1997, 105-176.

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 PP29

THE COMPARISON OF THE OCCURRENCE OF MALIGNANT UROLOGIC TUMORS IN TWO EIGHT-YEAR PERIODS (1980-1987 COMPARED TO 1998-2005) AT THE LJUDEVIT JURAK UNIVERSITY DEPARTMENT OF PATHOLOGY

 A. Dubravić1, T. Leniček1, K. Tomić2, M. Ulamec1, B. Krušlin1, M. Belicza1

 1Ljudevit Jurak Department of Pathology, Sestre milosrdnice University Hospital, Zagreb;
2Department of Pathology, General Hospital Dr. Josip Benčević, Slavonski Brod, Croatia

             AIM OF THE STUDY: According to the department’s cancer registry data in the last eight years urological cancers represented 32.2% (3015 cases) of all male cancers, and 8.7% (594 cases) of all female cancers.

The aim of this study is to determine the frequency and the pattern of urologic cancers diagnosed at our department in two eight-year periods a decade apart.

            MATERIALS AND METHODS: Histopathological data for the two periods were obtained from the department’s computer-based carcinoma registry, former being a period of 1980-1987 and the latter of 1998-2005. Patients were assessed according to the sex and the localization of the tumor.

            RESULTS: 1459 urologic cancers were reported over a period of 1980-1987 (83.6 % in males, 16.4% in females), opposed to 3609 cases (83.5% in males, 16.5% in females) diagnosed in the period of 1998-2005. The organ-specific distribution in males, in the former period, was as follows: urinary bladder 62.5%, prostate 24.7%, kidney 7.5%, testis 3.9% and finally, renal pelvis and ureter with 1.4%. In the latter period prostate cancer (diagnosed in 42.6% of cases) switched places with urinary bladder carcinoma (diagnosed in 36.9% of cases), with the remaining distributions’ ranks staying the same. One must emphasize that the frequency of prostate cancer rises noticeably in the last six years. In females a predominance of bladder tumors was found in both periods (74.2% of all urinary cancers in the first and 59.4% in the second period), followed by kidney and renal pelvis and ureter malignancies.

            DISCUSSION AND CONCLUSION: The absolute number of all urinary carcinomas increased three to four times during the observed periods, with the absolute number of prostate cancer quadrupling, mostly due to more sophisticated diagnostic methods. The bladder cancer is the most common urinary tumor in females, followed by kidney tumors showing slow but steady absolute and relative increase.

 References:

1.       PLESKO I. et al. Neoplasma 2004;51:248-54.

5. TRETARRE B. et al. Prog Urol 2003;13:394-403.
6. FISCHER CG. et al. Cancer 1998;82:775-83.

 

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PP30

MICROPAPILLARY SUBTYPE OF URINARY BLADDER CARCINOMA – A REVIEW OF TEN CASES

 M. Perić Balja1, T. Bujas2, K. Tomić3, T. Leniček4, M. Sučić5, B. Krušlin4

 1Department of Pathology, University Hospital for Tumors, Zagreb, Croatia; 2Department of Pathology, General Hospital Karlovac, Croatia; 3Department of Pathology, General Hospital Dr. Josip Benčević, Slavonski Brod, Croatia; 4 Ljudevit Jurak University Department of Pathology, Sestre Milosrdnice University Hospital, Zagreb, Croatia; 5Department of Urology, Sestre Milosrdnice University Hospital, Zagreb, Croatia

 
AIM: Micropapillary carcinoma represents an uncommon variant of urothelial carcinoma associated with higher grade and advanced stage at time of diagnosis, as well as with poorer prognosis. Even though the morphology may be deceptive, this tumor type is considered to be a tumor with an aggressive behavior. Aim of this study was to review patients with micropapillary variant of urothelial carcinoma in our database and compare them with patients with “classical” urinary bladder carcinoma.

MATERIALS AND METHODS: We used the histopathological database (Thanatos) from the Ljudevit Jurak Department of Pathology for all urologic patients with diagnosed urinary bladder cancer, during the time period from January 1st, 2004 to March 31st, 2006.

RESULTS: In the examined period there were 390 patients with urinary bladder cancer (M:F=298:92). Male patients were ageing from 26 to 91 years (mean 68.1) while females were ageing between 35 and 90 (mean 69.0). From the total number of patients with urinary bladder cancer there were only 10 (2.6%) cases with the micropapillary variant (M:F=8:2). In the group of patients with micropapillary carcinoma, at the time of diagnosis, males were ageing between 57-88 years (mean 70.9), while females were in age 76 and 81 years (mean 78.5). All micropapillary carcinomas were invasive at the time of diagnosis, and in 60% of all cases infiltration of muscular layer of urinary bladder was present. In 3 (30%) cases, after the diagnosis of micropapillary carcinoma, radical cystectomy was performed followed by lymphadenectomy, while only 7.9 % (30) cases with “classic” urothelial carcinoma were treated with radical operation. Lymph nodes metastases were confirmed histologically in all 3 cases with micropapillary cancer and in 38.1% (8/21) cases with classical urothelial cancer.

CONCLUSION: In our series micropapillary carcinoma was an aggressive tumor, often in advanced stage of disease, with a high incidence of lymph node metastases. Therefore, correct diagnosis is important to perform adequate, usually radical treatment.

 References

1.  AMIN MB et al. Am J Surg Pathol 1994;18:1224-32.

2.  DHOUIB RS et al. Pathologica 2005; 97:338-40.

3.  EBLE JN et al. Tumours of the Urinary System and Male Genital Organs,W Classification of Tumours, 2004

 

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PP31

TOPOISOMERASE IIα AND COLLAGEN IV EXPRESSION IN CARCINOMA OF THE UTERINE CERVIX

 Š. Smrkolj, M.Eržen, S.Rakar

 Department of Gynecology and Obstetrics, University Medical Centre Ljubljana, Slovenia

 AIM OF THE STUDY: The aim of this study was to analyze the immunohistochemical expression of topoisomerase IIα (topoII) and collagen IV in cervical cancer.

METHODS: A series of 114 cervical carcinomas were analyzed using standard IHC staining (IHS) procedures for topoII and collagen IV. The intensity and percentage of nuclear IHC staining was graded into four categories: negative, slight, moderate, intensive. Mean patients’ age was 42,3±11,5 years.

RESULTS: A high IHC expression of topoII was present in 55.3% of cases, and collagen IV in 28.1%. Intensive IHS for collagen IV was associated with weakly IHS to topoII. The IHS reaction to topoII was significantly decreased in adenocarcinoma compared to intensive IHS in squamous-cell carcinoma and was significantly associated to nuclear grade (p=0,038) and defense reaction (p=0,001). In the multivariate analysis the intensity of IHS to collagen IV was associated significantly with lymphovascular invasion (OR=5.906; 95%CI 2.18-15.96). The Kaplan-Meier analysis showed no significant differences in survival with regard to the intensity of IHC expression of topoII and collagen IV.

CONCLUSIONS: IHC expression of topoII and collagen IV is significantly correlated with the defense reaction. The survival analysis did not show significant differences with regard to expressions of topoII and collagen IV. The two markers of IHC indicated the existence of relevant factors at the molecular level that might complement the assessment of the prognosis of cervical cancer, resulting in the appropriate modification of the type and extension of cervical cancer treatment.

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PP32

MESONEPHRIC ADENOCARCINOMA OF UTERINE CERVIX- A CASE REPORT

 A. Škrtić1, A. Borovečki1, M. Milković Periša1, K. Horvat1, S. Džebro1, V. Kukura2, S. Gašparov1

 1Department of clinical pathology and cytology, 2Department of gynecology and obstetrics, University Hospital Merkur, Zagreb, Croatia

 Malignant mesonephric tumor of uterine cervix arising from mesonephric remnants is a very rare type of adenocarcinoma. The tumor is often located in the lateral and posterior wall of the cervix but may involve the cervix circumferentially as well. Morphological diagnosis is difficult because there are no characteristic immunohistochemical markers for this entity, especially for distinction from diffuse form of florid mesonephric hyperplasia.

A 53 years old female patient underwent hysterectomy and left adnexectomy due to hypermenorrhea and myomatous uterus. The uterine cervix was circumferentially and diffusely enlarged. Small tubular glands with intraluminal eosinophilic secretion, focally cystic, or branching tubules infiltrated circumferentially nearly the entire uterine cervix and isthmus, but without involvement of uterine corpus, surgical resection margins nor endocervical surface.

Neoplastic cells were immunoreactive with CK AE1/AE3, CK7, EMA, Ber-EP4, CD10, E- cadherin, p53; and negative for CK20, calretinin, alfa- inhibin, SMA, and vimentin. The tumor was negative for estrogen, progesterone, and androgen receptors. Ki-67 proliferation index in neoplastic cells was 8%.

Differential diagnosis included diffuse form of florid mesonephric hyperplasia and tubular morphologic pattern of mesonephric adenocarcinoma.

According to gross, morphologic finding, and phenotype profile of neoplastic cells we have decided that this is a case of adenocarcinoma of mesonephric type of uterine cervix.

 References

1. SILVER S.A. et al. Mesonephric adenocarcinomas of uterine cervix. Am J Surg Pathol2001;25:379-387.
2. ORDI J. et al. Mesonephric adenocarcinoma of the uterine corpus. Am J Surg Pathol 2001;25:1540-45.
3. BAGUE S. et al. Malignant mesonephric tumors of the female genital tract. Am J Surg Pathol2004;28:601-7.

 

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PP33

THE PROLIFERATIVE ACTIVITY OF ENDOTHELIAL CELLS ASSESSED BY IMMUNOHISTOCHEMICAL EXPRESSION OF Ki67 AND PCNA IN SOFT TISSUE TUMORS – ASSOCIATED ANGIOGENESIS

 Marius Raica1, Anca Maria Cimpean1, Virginia Martin2, Ovidiu Mederle1

 1Victor Babes University of Medicine and Pharmacy, Department of Histology&Cytology, Timisoara, Romania, 2 Clinic Hospital, Department of Pathology, Arad, Romania

 BACKGROUND: Angiogenesis is a process of new blood vessels formation from preexisting ones. Tumor associated angiogenesis is currently investigated through microvessel density (MVD), but MVD tells almost nothing about the tumor responsiveness to antiangiogenic therapy. Our purpose was to investigate proliferative activity of endothelial cells in the tumor area.

MATERIAL AND METHODS: We investigated 44 patients with primary soft tissue sarcoma. After pathologic diagnosis and immunophenotyping we discovered 4 fibrosarcoma, 6 leiomyosarcoma, 2 rhabdomyosarcomas, 10 liposarcoma, 14 malignant fibrous histiocytoma (MFH), 4 myxoid sarcoma, and 4 pleomorphic sarcoma. Slides from each case were stained for CD34, CD31, PCNA, and Ki67; we used the LSAB2/EnVision operating system, and DAB was used as chromogen. MVD was calculated in each case, using the hot spot method. Observations were focused on blood vessels with endothelium positive for both CD34 and CD31, and aspects were compared with results obtained with proliferation markers.

RESULTS: Blood vessels of all tumors were stained for CD34, 38 cases were, also, stained with anti-CD31. High value of MVD was found in liposarcoma and MFH (mean average 43.8/x200). MVD did not correlate with the outcome of the patient or grade of the tumor. Blood vessels in tumor area stained with PCNA and Ki67 showed activated endothelial cells only in vessels devoid of perivascular cells. Mature blood vessels were consisted of endothelial cells that were negative for both markers, but positive reaction was noticed in pericytes/smooth muscle cells. Based on this, tumor associated blood vessels were classified as immature, intermediate, and mature.

CONCLUSION: The proliferative activity of endothelial cells in blood vessels associated to the soft tissue sarcoma was demonstrated through PCNA and Ki67 in immature and intermediate vessels, which were presumed to be the main target for potential antiangiogenic therapy.

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 PP34

CYTOLOGICAL FINDINGS IN A CASE OF PRIMARY PULMONARY PLASMACYTOMA (PPP)

 E. Garcia-Ureta, E. Carro Rey, O. Robles Viega, R. Alvarez Rodriguez

 Citologia Servicio De Anatomía Patológica Hospital Universitario Juan Canalejo, La Coruña, Spain

             OBJETIVE: Extramedullary plasmacytoma is a rare immunoproliferative monoclonal disease and only a few cases diagnosed by cytological studies have been reported. It has to be differentiated from other neoplasm and reactive plasma cell proliferations. Cytological findings of bronchial brushing and washing of PPP are shown in the case report.

            CASE REPORT: A 58 year-old man was hospitalized for assessment of a right hilar mass that was noted on a chest radiograph; laboratory data were all within normal limits. Material for cytological and histological examination was obtained during fiberoptic bronchoscopy from the endobronchial lesion. Brushing and washing cytological samples were processed by conventional preparations, Thin-Prep technique, staining techniques and immunocytochemistry.

            RESULTS OF CYTOLOGICAL FINDINGS: Cytological examination reveals a dispersed population of cells with a plasmacytoid appearance with eccentric nuclei, abundant cytoplasm and the characteristic paranuclear hof. Binucleation was common and mitotic figures were easily identified. Inmunocytochemical stains revealed that the tumor cells stained for kappa light chain, the cytological diagnosis was plasmacytoma and was confirmed histologically. Work up to rule out multiple myeloma was completed. Skeletal survey showed no other sites of involvement. Serum electrophoresis was normal and Bence Jones proteinuria was absent. Bone marrow examination demonstrated no abnormalities. Based on these findings an extramedulary plasmacytoma of the lung (PPP) was diagnosed.

            CONCLUSIONS: Cytology material of brushing and washing the affected area can be a safe and fast procedure for diagnosing myeloma of the lung. Mainly, these tumors need to be distinguished from reactive inflammatory processes, plasmacytoid lymphoma and plasma cell granuloma.

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 PP35

FETAL SURVIVAL DETERMINED BY 5-AZACYTIDINE IMPACT ON THE PLACENTA

 Lj. Šerman1, N. Sinčić1, M. Vlahović1, F. Bulić-Jakuš1, G. Jurić-Lekić2, A. Šerman3, A. Katušić1

 1Department of Biology, 2Department of Histology and Embryology, School of Medicine, Zagreb, Croatia; 3University hospital Sveti duh, Zagreb, Croatia

DNA methylation as a regulatory mechanism for mammalian gene expression is crucial for normal placentation as well as fetal development. Investigation of DNA methylation is performed by applying a demethylating agent 5-azacytidine to pregnant rats. 5azaC incorporates into the DNA instead of cytosine and prevents its methylation (Shiota K, et al. Genes Cells 2002; 7: 961), subsequently it changes expression of genes involved in normal placental and fetal development.

Single dose of 5azaC (5 mg/kg) was administered to pregnant rats at different days, namely 11th, 12th and 13th, of gestation. Placentas and fetuses were isolated at day 20, weighted and histologically analyzed.

After administering 5azaC on the 11th day of gestation, treated placentas were significantly smaller. Labyrinth, as a functional main part of the placenta, was reduced and consequently complete fetal resorption occurred. The same agent administrated at day 12 of gestation resulted in 24% of fetal survival, while after its application only one-day later (13th day of gestation) survival was 96%. Placentas treated on day 12 of gestation demonstrated a slightly recovered labyrinth, while placentas of day 13 showed almost normal distribution of placental layers.

These results confirmed epigenetic influence upon placental development. 5azaC caused changes in its structure, especially the reduction of labyrinthine part that is crucial for fetal survival. After establishment of normal placental layers distribution, 5azaC has no impact on placental morphology, neither on fetal survival. On the other hand, the influence of 5azaC remains, visible in appearance of fetal malformations (Sinčić N. et al. Period biol 2002; 104: 441).

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 PP36

CD105/SMOOTH MUSCLE ACTIN DOUBLE IMMUNOSTAINING DISCRIMINATE BETWEEN IMMATURE AND MATURE TUMOR BLOOD VESSELS

Anca Maria Cimpean, Marius Raica, Cristian Suciu

 Victor Babes University of Medicine and Pharmacy, Department of Histology and Cytology, Timisoara, Romania

 AIM OF THE STUDY: to demonstrate the value of the double immunostaining for endothelial and perivascular cell to discriminate mature from immature tumor-associated blood vessels in mammary carcinoma.

MATERIAL AND METHODS: 22 patients with ductal invasive mammary carcinoma were investigated. Tissue specimens were fixed in buffer formalin, embedded in paraffin and sections were stained with haematoxylin-eosin. Additional sections 5μm thick were performed for immunohistochemistry, in order to demonstrate the type and incidence of newly formed blood vessels in the tumor area, classified according to the Gee’s system. We used the specific endothelial marker CD105 (endoglin) to highlight activated endothelial cells and antibody against smooth muscle cell actin (SMA) for perivascular cells, applying Envision Doublestain system (HRP/DAB-APAAP/Fast Red). Both CD105 positive and SMA positive blood vessels were counted using the hot spot method, assisted by Lucia G microscopic image analysis program.

RESULTS: We found an inverse correlation between the expression of CD105 and SMA in vessels located outside the tumor area. We noticed a strong positive signal for CD105 in the intratumor single endothelial cells and intermediate vessels. Only few blood vessels were positive for both CD105 and SMA within the tumor. As the signal for the endothelial marker decreased or became negative, the expression for SMA increased. Using the double immunohistochemical staining method, we found 37% of immature vessels (single endothelial cells), 41% of intermediate, and 12% of mature (with pericyte/smooth muscle cells) in the tumor area. Outside the tumor area the values were 4%, 14%, and 82%, respectively.

CONCLUSION: Our results demonstrate significant predominance of immature and intermediate blood vessels in the tumor area of the mammary carcinoma. The differentiation between vessels with and without perivascular cells coverage using double immunostaining for CD105 /SMA may be an important step in the selection of patients which could benefit from antiangiogenic therapy with monoclonal antibody against CD105 antigen.

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 PP37

SECONDARY LEIOMYOMATOSIS IN A BOSNIAN MALE – A CASE REPORT

 Semir Vranić1, Nurija Bilalović1, Zinaida Ćerimagić2, Antonietta Gatti3, Klaus Kayser4

 1Department of Clinical Pathology and Cytology, Clinical Center of the University of Sarajevo, Sarajevo, Bosnia and Herzegovina; 2Department of Thoracic Surgery, Clinical Center of the University of Sarajevo, Bosnia and Herzegovina; 3University of Modena and Reggio Emilia, Modena, Italy; 4UICC-TPCC, Institute of Pathology, Charite, Berlin, Germany

 We describe an interesting case of chronic allergic pneumonia with severe interstitial fibrosis and secondary leimyomatosis in a 37 year-old Bosnian male who was wounded in 1993, during the war in Bosnia. The patient was admitted at the Department of Thoracic Surgery in January 2006 due to progressive respiratory problems. The clinical diagnosis was suspected inflammatory cystic changes of the lung. The surgical procedure was performed followed by histopathological evaluation. The specimen showed progressive interstitial fibrosis (Vimentin positive), disturbed lung structure with squamous cell metaplasia and moderate atypia (CK7 positive), hyperplastic bronchial mucosa, numerous eosinophils with obliterative small vessels and broad areas with proliferating smooth muscle cells (SMA positive). These changes were consistent with the above-mentioned diagnosis and were probably caused by the war trauma of the lungs. Immunohistochemical evaluation for estrogen and progesterone receptors was negative.

Further diagnostic procedures (detection of residual nanoparticles) will be performed in order to clarify potential causes of such complex pulmonary changes.

 

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PP38

MUTILATION OF THE HUMAN BODY

 M. Marcikić, B. Dumenčić, Elizabeta Matuzalem

 Medical Faculty, Department of Pathology and Forensic Medicine, Osijek, Croatia

 Mutilation of the human body is a phenomenon documented from ancient times. It is one of the darkest traits that can occur in a human being. The cases were classified into four groups, with regard to the primary motives that were considered to underlie the mutilation. The first group covered defensive mutilation, where the motive is to get rid of the body and/or make its identification more difficult. This type is also denoted as dismemberment. The second one was defined as aggressive mutilation, where the act of killing is brought by a state of outrage, and is followed by mutilation of the body. And the last one was offensive mutilation, necrosadistic murders, motivated by a necorphilic urge to kill and to carry out sexual activities with a dead body, with prior or subsequent mutilation, or a sexual sadistic need to carry out sexual activities while inflicting pain or injuries, or killing and where the mutilation may be initiated in a living person and continued after the killing, or may be commenced after the killing. Necromaniac mutilation, carried out on a dead body. This is sometimes seen in regular necrophilia, as defined above, or with a purpose of using some body parts as a trophy, symbol or fetish. We had been presented with bizarre pattern of killing associated with mutilation of a dead body. The body of a 67 year old lady had been found dismembered in her house. The event had occurred in January 2005. The corpse was decapitated, arms and legs amputated. Abdominal and thoracic cavities had been cut wide open and organs removed with a knife. Pieces of body parts were put in the toilet and the bathtub. In spite of the gross mutilation of the body which may cover the cause of death, evidence of blunt vital craniocerebral injuries were confirmed. Facial destruction prevented visual identification of the dead body. The final identity of the body was verified by dactiloscopy. The killer was the 29 year old lady’s son. He was an introverted, lonely man with history of mental health problems. Neighbors called the police alarmed by the noise coming from the lady’s house. The perpetrator stayed in the house until the police arrived. The local community was appalled with what happened in their vicinity. This eccentric killing coupled with gross mutilation of the victim’s body raised many questions about the motivation, stress factors and personality of the killer. Mutilation of this type has been reported in cases where the perpetrator and the victim have been closely related or where the motive for the killing was revenge. Common pattern in these cases was the presence of multiple injuries indicating a prompt action. Psychotic perpetrators with schizophrenia or affective disorders were among those in this group. These were basic elements of the aggressive mutilation as was our case. Questions whether such horrible familiar tragedy could be foreseen and prevented or not were frequently asked in the aftermath of the event.