Prognostic value of survivin expression in Wilms tumor.

PURPOSE: To determine survivin expression patterns in Wilms tumor (WT) and compare it with the expression in normal renal tissue. Also, to analyse cytoplasmic and nuclear survivin expression in relation to histological type, prognostic group and tumor stage. METHODS: Immunohistochemical expression of survivin was analysed in 59 cases of primary WT and in 10 normal kidney specimens, taken from the same patients, but distant from the tumor. RESULTS: 51 out of 59 cases of WT (86.44%) showed decreased cytoplasmic survivin expression and 4 out of 59 cases of WT (6.78%) showed nuclear overexpression of survivin. There was statistically significant difference in the frequency of decreased cytoplasmic expression of survivin in individual components of WT (p=0.005). Decreased cytoplasmic expression of survivin in epithelial, blastemal and stromal component was found significantly more often in low stage WT compared to high stage WT (Fisher exact test, p=0.0002, p=0.002, p=0.002, respectively). There was no statistically significant difference in the frequency of survivin nuclear overexpression between different stages of WT (Fisher exact test, p=0.564), histological types (Fisher exact test, p=0.915), or between different prognostic groups (Fisher exact test, p=1). CONCLUSION: Decreased survivin cytoplasmic expression or nuclear overexpression may be related to favorable prognosis of WT.

Prognostic significance of clinical parameters and Epstein-Barr virus infection in non-endemic undifferentiated carcinoma of nasopharyngeal type: a Serbian report.

Undifferentiated carcinoma of nasopharyngeal type (UCNT) is very rare tumour in Serbia, like in most of the countries of Europe, with incidence less than 0.5 per 100,000 people per year. The aim of this study was to assess the presence of Epstein-Barr virus (EBV) in the UCNT of a non-endemic population in Serbia and identify the main clinical parameters that interfere with patients' survival rate. This study included 102 patients with UCNT who were diagnosed between 1996 and 2003. Biopsies were analysed for EBV-encoded RNA (EBER) by in situ hybridization of tumour tissue microarray. Of 102 patients, 76 were men and 26 were women with ages ranging between 18 and 82 years (median 52.5, mean 53.0±14.1). Survival rates were 80, 39 and 31% for one, three and five years, respectively. Ninety-three of 102 cases were EBER positive (92%). Factors with unfavourable prognostic values were age over 50 years at the time of diagnosis, advanced clinical stage, therapy other than chemoradiotherapy and EBER negative status. In regard to the clinical data, EBER expression in UCNT was shown to be a strong independent predictor of overall and progression-free survival. To our knowledge, the current report constitutes the largest European non-endemic series of UCNT samples from a single institution with correlation between survival and clinical parameters/EBER status.

Large-cell neuroendocrine carcinoma of the ampulla of Vater.

Large-cell neuroendocrine carcinoma is a high-grade neuroendocrine carcinoma, originally described in the lung. The tumor rarely occurs in extrapulmonary sites like the gastrointestinal tract, and only few examples have been described in the ampulla of Vater. A new case of large-cell neuroendocrine carcinoma of the ampulla of Vater in a 60-year-old man is reported. After pancreatoduodenectomy, macroscopic examination revealed ulcerated tumor in the region of the ampulla of Vater. Microscopically, the tumor exhibited organoid, predominantly nested growth pattern, consisting of large, polygonal cells with pleomorphic nuclei. Average number of mitoses was 36 per 10 high-power fields. Small and large areas of necrosis were identified. Immunohistochemically, the tumor cells were positive for synaptophysin, chromogranin A, PGP 9.5, neuron-specific enolase, pancytokeratin, CK8 and somatostatin and negative for CK7, CK20, S-100, TTF-1, HMB-45, CD117, E-cadherin and regulatory peptides. Ki-67 proliferative index was 41%. Histone deacetylase (HDAC) analysis showed almost identical results for HDAC1, HDAC2 and HDAC3--60, 60.3 and 61%, respectively. Two months after surgery, liver metastases occurred, confirming highly aggressive behavior of large-cell neuroendocrine carcinoma.

Changes in common melanocytic naevi after intense sun exposure: digital dermoscopic study with a 1-year follow-up.

BACKGROUND: Exposure to ultraviolet (UV) radiation induces acute changes in common melanocytic naevi (CMN). AIM: To analyse changes in size and dermoscopic structures of CMN in healthy individuals under the influence of intense sun exposure and to investigate effectiveness of sunscreen usage in preventing the appearance of dermoscopic changes. METHODS: The subjects were divided into an SS group and a non-SS group, according to whether they used sunscreen or not. Digital epiluminiscence microscopy (DELM) was performed before sun exposure and 28 days and 1 year after the cessation of intense sun exposure. RESULTS: Eleven subjects with 60 naevi fulfilled the inclusion criteria. A significant increase in one diameter of the CMN was found 28 days after cessation of sun exposure, irrespective of sunscreen usage. There was no significant change in total dermoscopy score during the follow-up period in either the SS or non-SS group. There were definitive DELM changes at the 1-year follow-up in 10% of CMNs < 5 mm in size. Changes in pigmented network, globules and streaks were fully reversible in the remaining CMN. The use of sunscreen had no influence on these changes. CONCLUSIONS: The increases in the size of melanocytic naevi that we found 28 days after cessation of intense sun exposure are probably induced by UV radiation, whereas changes in DELM features that were not reversible in some small naevi at the 1-year follow-up could be related both to sun exposure and to the natural evolution of the naevi. The use of sunscreen does not seem to prevent these changes. Small naevi in low-risk young and middle-aged patients seem to be more prone to sun-induced DELM changes.

[The excision width in surgical treatment of basal cell carcinoma].

Basal cell carcinoma originates from pluripotent cells of basal layer of epiderm, external covering of hair follicles, sebaceous glands or other skin adnexa. It is characterized by local infiltrating and sometimes destructive growth. There are several types of basal cell carcinomas that may be manifested in over 12 clinical forms. Surgical treatment depends to a large extent on the histological type, localization and its clinical manifestation. The analysis included 250 patients of both gender and different age, operated for basal cell carcinoma. Clinical characteristics of basal cell carcinoma and the width of the excision were described. It was concluded that the width of the excision of basal cell cancer was in relation to histological type.

Cyclin A and beta-catenin expression in actinic keratosis, Bowen's disease and invasive squamous cell carcinoma of the skin.

BACKGROUND: Actinic keratosis (AK) has been defined as a precancerous lesion or an early phase in the evolution of squamous cell carcinoma (SCC) and histological changes seen in the individual cells of an AK are indistinguishable from those seen in SCC, which invade the dermis. Cyclin A is an increasingly utilized proliferation marker that has functions in both S phase (DNA replication) and initiation of mitosis, whereas alterations of beta-catenin, the molecule involved in cell-cell adhesion and in signalling transduction, could promote invasive and proliferative capacities of malignant tumours. OBJECTIVES: To determine cyclin A and beta-catenin expression pattern in cutaneous SCC and in in situ lesions classified as keratinocytic intraepidermal neoplasia (KIN) and, using traditional terms, as AK and Bowen's disease (BD), and to analyse it in relation to SCC differentiation, diameter and thickness. METHODS: Immunohistochemical staining was performed on 110 formalin-fixed paraffin-embedded tissue samples with the streptavidin-biotin technique using antibodies to cyclin A and beta-catenin. On histological examination, 53 lesions were diagnosed as AK, 16 as BD and 41 as SCC-11 well differentiated (WD), 16 moderately differentiated (MD) and 14 poorly differentiated (PD). Using KIN classification, 22 lesions were KIN1, 23 were KIN2 and 24 were KIN3. For cyclin A, distribution and labelling index (LI), and for beta-catenin, level of membranous staining and presence of aberrant (nuclear/cytoplasmic) localization were examined. RESULTS: Diffuse cyclin A presence was observed more frequently in BD than in AK (P < 0.0001) or SCC (P = 0.0002), and in SCC-PD compared with SCC-WD (P < 0.0001) or SCC-MD (P = 0.0003). Differences between KIN3 and KIN2, as well as KIN3 and KIN1 lesions, were statistically significant (P < 0.0001), and the same result appeared when KIN1 and KIN2 cases were grouped and compared with those of KIN3 (P < 0.0001). Cyclin A LI was significantly lower (P < 0.05) in AK than in BD or SCC, but no difference between BD and SCC was found, and LI in BD was even higher than in SCC-WD or SCC-MD, while analysis regarding SCC differentiation and KIN classification revealed the same correlation as for the cyclin A distribution. Reduced or absent beta-catenin membranous staining was found in 90 cases (81.8%), more often in SCC than in AK (P = 0.03) or in AK and BD grouped together (P = 0.02). There was no statistical difference between SCCs of various level of differentiation, or between different KIN grades. Diffuse loss of membranous beta-catenin staining showed 36 lesions (32.7%), more frequently SCC than AK (P = 0.003) or AK and BD grouped (P = 0.006), as well as SCC-PD compared with SCC-WD (P = 0.01) and SCC-MD (P = 0.03), whereas all KIN comparisons remained nonsignificant. Aberrant beta-catenin cellular localization demonstrated 28 lesions (25.5%), most often in the basal or peripheral parts and in the lesions with diffuse beta-catenin loss (P = 0.009), but revealed no correlation with the histological type, SCC level of differentiation or KIN grades. Diffuse loss of membranous beta-catenin staining was found to be significantly more frequent in SCC thicker than 4 mm (P = 0.03), while all other comparisons between cyclin A or beta-catenin with the tumour size remained nonsignificant. Cyclin A LI was higher in cases with diffuse loss of membranous staining (P = 0.001) or with aberrant cellular localization of beta-catenin (P = 0.002). CONCLUSIONS: Cyclin A LI showed greater difference between AK and BD than between BD and SCC, suggesting that increased proliferation (measured by cyclin A LI) characterizes progression of in situ lesions from AK to BD, whereas reduced beta-catenin expression separates more clearly SCC from the in situ lesions. Diffuse pattern of loss of membranous beta-catenin staining correlated better with the type of lesion, SCC differentiation and tumour size than reduced expression in general or aberrant cellular localization of beta-catenin. KIN classification does not seem to be supported by our findings, except when KIN1 and KIN2 lesions (in situ, partial thickness) are grouped.

Umbilical metastasis (Sister Joseph's nodule) as a first sign of a disseminated ovarian carcinoma: comparative immunohistochemical analysis of primary tumor and its metastases.

The case of a 46-year-old female with umbilical metastasis as a first sign of an ovarian carcinoma is reported with the results of immunohistochemical analysis of primary tumor and lymph node and umbilical metastases. All specimens were positive for cytokeratin 7, CA 125, E-cadherin, alpha-, beta-, and gamma-catenin, as well as for MSH2. Staining with cytokeratin 20 and MLH1 was negative, and Ki-67 labeled from 5% (in the center of the lesions) to over 25% (at the periphery of the lesions) of the nuclei. Beta-catenin showed membranous positivity in the central parts and absence of staining at the periphery of ovarian tumor and umbilical metastasis, whereas lymph node metastasis presented with uniform reaction throughout. The results of immunohistochemical staining could point to the mechanisms employed by malignant tumors during invasion and growth of metastasis and suggest the possible role of the microenvironment in the expression of some adhesion molecules on tumor cells.

Giant nevus lipomatosus superficialis with multiple folliculosebaceous cystic hamartomas and dermoid cysts.

Nevus lipomatosus superficialis (NLS) is a rare benign condition characterized by papules and nodules usually in the pelvic and sacral areas, with ectopic mature adipose tissue reaching the superficial dermis. We report the case of a 47-year-old female with large NLS in the sacral region that had been present since birth and, after an asymptomatic course for most of the time, became associated with episodes of mild pain that prompted the patient to look for medical help. After a complete resection of the cerebriform plaque, measuring 23 cm in diameter, histopathological examination revealed typical NLS architecture in as yet undescribed association with multiple folliculosebaceous cystic hamartomas (FCHs) and dermoid cysts.

Cadherins and integrins in renal cell carcinoma: an immunohistochemical study.

AIMS AND BACKGROUND: The aim of this study was to determine the expression of cadherins and integrins in renal cell carcinoma (RCC) and their relationship with tumor morphology and TNM status. METHODS: Cadherin and integrin expression was investigated using an indirect immunoperoxidase technique, applying antibodies to E-, N-, P- and VE-cadherin and to alpha1, alpha2, alpha3, alpha4, alpha5, alpha6, and alpha(v) integrin subunits. Correlation of semiquantitatively scored adhesion molecule levels with histopathological parameters (cytology, growth pattern, nuclear grade) and TNM status was performed for 24 RCCs (17 clear cell, 3 granular, 3 spindle cell and 1 chromophobe cell type according to the WHO classification). RESULTS: E-cadherin and N-cadherin were present in most cases (88% and 67%, respectively) and were usually coexpressed. T3 RCCs displayed higher E-cadherin and N-cadherin levels than T1/T2 tumors regardless of tumor grade, suggesting that impairment of their function might exist without actual loss from tumor cells. P-cadherin was found focally in two RCCs only, while VE-cadherin was present on stromal vessel endothelium in five tumors, showing no differences with regard to cell type, growth pattern, tumor grade or TNM status. All integrins were present in the studied RCCs (ranging from 12% for alpha5 to 79% for alpha3), including those that are normally absent from adult kidney tissue (alpha4 and alpha5). Tumors of higher grade showed increased alpha(v) and decreased alpha6 levels, while RCCs with metastases less often showed diffuse alpha3 presence and never expressed alpha5 integrin. CONCLUSIONS: Our results suggest that the level of expression of N-cadherin and some integrins (most notably alpha3, alpha6 and alpha5) is associated with the capacity of RCC for local and distant spread, regardless of tumor grade.

Immunohistochemical analysis of HLA class II antigens and tumor infiltrating mononuclear cells in renal cell carcinoma: correlation with clinical and histopathological data.

Cryostat sections of 37 renal cell carcinomas (RCC)--25 clear cell type, 10 granular and 2 chromophobe--were studied with indirect immunoperoxidase method applying monoclonal antibodies (MoAb) to HLA-DR, -DP and -DQ antigens for analysis of HLA class II antigens, and anti-CD14, -CD3, -CD4 and -CD8 MoAb for tumor infiltrating mononuclear cells (TIM). Number of positive cells was estimated semiquantitatively and results of immunohistochemical investigation were correlated with clinical (patient age and sex, tumor size and TNM stage) and histopathological (cytology, histology, grade) characteristics of RCC. All RCC expressed HLA-DR, 92% -DQ and 73% -DP antigens with level of expression in hierarchy- DR>-DQ>-DP, but no statistically important correlation could be established with any of the histopathological or clinical parameters analyzed. Monocytes were more abundant than T lymphocytes and CD4+ than CD8+ T cells, whereas tumors with T lymphocyte predominance and approximately equal number of CD4+ and CD8+ T cells had greatest average diameter. Inadequate activation of T lymphocytes by tumor cells (despite capability of antigen presentation) could be the reason for association of parameters which indicates more aggressive tumor behavior with aberrant HLA class II antigen expression on RCC.

HLA class I antigens expression in renal cell carcinoma: histopathological and clinical correlation.

Immunohistochemical analysis of HLA class I antigens expression in 26 renal cell carcinomas (RCCs)--18 clear cell, 6 granular and 2 chromophobe--was performed with indirect immunoperoxidase method. Results were correlated with extent and immunophenotype of tumor infiltrating mononuclear cells, histopathological (histology, cytology, grade, presence of necrosis) and clinical (tumor diameter, TNM classification) characteristics of RCC. 4 (15%) RCCs showed reduced HLA class I presence and this was associated with greater tumor diameter and more frequent T3, T4 and M1 stage. All tumors with altered HLA class I antigens expression were grade 2 or 3 and strong correlation with presence of necrosis (p=0.006) was noticed, while mononuclear cell infiltrates (especially CD8+ T lymphocytes) were less extensive compared to tumors with normal HLA class I level. Our results suggest more aggressive clinical behavior of RCCs with reduced HLA class I antigens expression, probably due to impaired cellular antitumor immune response.

[Histogenesis of renal cell carcinoma]. / Histogeneza karcinoma bubreznih celija.

Immunomorphological characteristics of 27 renal cell carcinomas--18 clear cell, 6 granular, 2 chromophobic, 1 sarcomatoid--as well as 1 oncocytoma were analyzed. The investigation was performed on cryostat sections with indirect immunoperoxidase technique using monoclonal antibodies to intermediate filaments-cytokeratin and vimentin--and renal differentiation antigens--CD10 and CD24. All carcinomas, with the exception of chromophobic type, showed cytokeratin/vimentin coexpression together with strong CD24 and weak (or absent) CD10 staining indicating at primitive cells with initial differentiation toward proximal tubule epithelium as most probable site of origin. In chromophobic cells only cytokeratin and CD24 antigen presence was observed, pattern similar to that seen in oncocytoma. It could be supposed that those two tumors have closely related histogenesis, originating from more differentiated cells with tendency to develop toward distal tubule epithelium.

[Ultrastructural changes in transplantation glomerulopathy]. / Ultrastrukturne promene transplantacione glomerulpatije.

Twenty-eight biopsy specimens were obtained from patients 3-95 months after kidney transplantation and studied by light, electron and in some cases also by immunofluorescence microscopy. Electron microscopic studies showed that the most frequent glomerular lesion was widening of lamina rata interna which is accompanied with subendothelial accumulation of finely granular material, formation of new subendothelial basement membrane and deposition of microfibrils and fine filaments. The mesangial changes were mainly those of mesangiolysis and mesangial sclerosis with deposition of mesangial matrix and microfibrils, but little cellular proliferation. Fragmented red blood cells were seen in nearly half of the patients. Arterial intimal thickening and occasionally also thrombosis produced ischaemic changes in the kidney and in the glomeruli and contributed to the process of transplant rejection.

[Electron microscopic analysis of eosinophilic renal cell carcinoma]. / Elektronosko-mikrospopska analiza eosinofilnog karcinoma celija bubrega.

Renal cell carcinoma with cytoplasmic eosinophilic globules visualized on routine histologic preparations was analyzed. Eosinophilic globules in cytoplasm of the cells in renal cell carcinoma are very rate and till today we have not heard or found in the literature an attempt to analyze and describe them and that was the aim of our study. By electron microscopy, the globules most closely resembled non-membrane bound filamentous material that normally constitutes the cytoskeleton of normal and neoplastic renal epithelium.

[Intercellular adhesion molecule-1 expression in renal cell carcinoma]. / Ispoljavanje intercelulne adhezivne molekule-jedan u karcinomu celija bubrega.

The presence of intercellular adhesion molecule-1 (ICAM-1) was analyzed using indirect immunoperoxidase technique in frozen sections of 27 renal cell carcinomas (RCC)--18 clear cell, 6 granular, 2 chromophobe and 1 sarcomatoid. Great variations in ICAM-1 expression were observed in clear and granular cell RCC, without correlation with mononuclear (lympho-monocytic) cell infiltration. Sarcomatoid type displayed widespread ICAM-1 distribution and intense mononuclear infiltrate, while chromophobic RCC, despite better prognosis, expressed ICAM-1 weakly and focally, almost without mononuclear infiltrate. With such results, no relation between ICAM-1 expression on tumor cells and behavior of various RCC types could be established and possible explanations are: different histogenesis of some RCC (chromophobe), nonspecific upregulation of ICAM-1 as a result of ischaemic-necrotic processes, or negative effect of ICAM-1 expression on complement action.

Immunomorphological characteristics of renal cell carcinoma.

Immunomorphological characteristics of 27 renal cell carcinoma (RCC): 18 clear cell, 6 granular (chromophilic), 2 chromophobe, 1 spindle cell (sarcomatoid) as well as of 1 oncocytoma, were analyzed. The investigation was performed on cryostat sections by immunoperoxidase technique applying a panel of monoclonal antibodies which defined: proximal (TNE3, TN5, 5D9) and distal (TN8, TN9, 7C2) tubular antigens; intercellular adhesion molecule 1 (ICAM1); HLA class II (-DQ, -DR and -DP) antigens, intermediary filaments (cytokeratin and vimentin); and antigens on tumour infiltrating mononuclear leucocytes (TT1, TT2 and LeuM3 for CD4, CD8 and CD14 antigens, respectively). All RCC with exception of chromophobe co-expressed cytokeratin and vimentin. In addition, they were usually positive for all proximal and two distal tubular markers (TN8, TN9) indicating primitive cells which could differentiate into the epithelium of both parts of tubule system as the most probable originators of in RCC. Almost all RCC but the chromophobe aberrantly expressed HLA class II antigens which great variability from case to case. The presence of HLA-DR antigens was more intensive and widespread than of HLA-DQ and -DP antigens. Expression of ICAM1 mostly correlated with presence of HLA class II antigens, particularly with -DR on tumour cells of RCC. HLA-DR antigen expression was always more prominent than mononuclear cell infiltrate (among which macrophages prevailed over T cells) which could suggest that increased histocompatibility antigen expression precedes mononuclear cell influx. In contrast to all other RCC, chromophobe tumours had quite distinct features revealing the most intense reaction with 7C2 (MAb that produced the weakest reaction with other tumour types), absence of vimentin and very weak reaction with antibodies for HLA class II Ag and ICAM1.(ABSTRACT TRUNCATED AT 250 WORDS)

[Immunohistochemical analysis of malignant tumors of the thyroid gland using 6 relevant markers]. / Imunohistohemijska analiza malignih tumora stitaste zlezde primenom sest relevantnih markera.

The results of immunohistochemical study of malignant thyroid tumours using six markers-thyroglobulin (TGB), keratin, carcinoembryonic antigen (CEA), epithelial membrane antigen (EMA), calcitonin and leucocyte common antigen (LCA)-are presented. Thirty-three cases of thyroid carcinomas were studied: 11 papillary, 8 follicular, 7 medullary and 7 anaplastic carcinomas. All cases of papillary and follicular tumours showed positive reaction with TGB and 4 papillary and 3 follicular carcinomas reacted with CEA also. 100% reactivity with calcitonin and CEA were observed in the medullary carcinoma group, while anaplastic carcinomas revealed TGB (one case), CEA (two) and EMA (three) reactivity. None of the studied tumours responded to LCA. TGB and calcitonin proved to be useful in distinguishing tumours originating in follicular cells from those originating in parafollicular cells and, along with LCA, enabled discrimination of anaplastic carcinomas from medullary carcinomas and malignant lymphomas. CEA can be only used as an auxiliary marker in cases of medullary carcinoma, while EMA and high molecular weight keratins did not prove to be reliable markers for analysis of malignant thyroid tumours.