TGF-ß1 expression in chromophobe renal cell carcinoma and renal oncocytoma.

Distinguishing renal oncocytoma (RO) from the eosinophilic variant of chromophobe renal cell carcinoma (ChRCC) under the light microscope is a common diagnostic problem. Our recent research has shown significant difference between the presence of tumor fibrous capsule in ChRCCs and ROs. Transforming growth factor beta 1 (TGF-ß1) is a potent cytokine involved in regulating a number of cellular processes. Two main purposes of this research were to investigate whether the TGF-ß1 staining could be related to the presence of tumor fibrous capsule and if it could be used in the differential diagnosis between ChRCC and RO. We investigated 34 cases: 16 ChRCCs (8 eosinophilic and 8 classic) and 18 ROs. All available slides of each tumor, routinely stained with hematoxylin and eosin (H&E) were first analyzed to note the presence of tumor fibrous capsule. One paraffin embedded tissue block matching the representative H&E slide was selected for the immunohistochemical analysis. TGF-ß1 expression was analyzed semiquantitatively in the tumor tissue, the tumor fibrous capsule, if present and the peritumoral renal parenchyma. Intensity of TGF-ß1 expression was weaker in ChRCCs than the one observed in ROs (P<0.05). The type of reaction in ChRCCs was predominantly membranous unlike in ROs, which exhibited a predominantly cytoplasmic reaction (P<0.05). Moreover, none of the ROs showed membranous type of reaction for TGF-ß1. In the group of ChRCCs, tumors with capsule had statistically significant higher quantity of TGF-ß1 expression in tumor tissue and in peritumoral renal parenchyma compared to the tumors without capsule (P<0.05). Our results showed different types of TGF-ß1 expression in ChRCCs and ROs: ChRCCs had predominantly membranous type of reaction, and ROs predominantly cytoplasmic. Furthermore, ChRCCs with capsule had statistically significant higher quantity of TGF-ß1 expression in tumor tissue and in peritumoral renal parenchyma compared to the tumors without capsule. Based on these findings we can speculate that it could be possible that TGF-ß1 plays a role in the formation of fibrous capsule in ChRCCs.

Micropapillary urothelial carcinoma of the ureter.

Micropapillary urothelial carcinoma (MPUC) is a rare aggressive variant of urothelial carcinoma, associated with advanced tumor stage, high tendency to invade lymphovascular spaces, and metastasize to lymph nodes and other organs. Therefore, it has a poor prognosis. One of the most prominent histological features is the presence of small, round empty spaces surrounding infiltrating tumor nests. If detected, even a small focus of micropapillary pattern may be therapeutically significant; the higher proportion of micropapillary component, the worse the prognosis. Radical nephroureterectomy is the treatment of choice even in the setting of superficially invasive disease. Although, MPUC has been well studied in urinary bladder, only a few cases of MPUC in upper urinary tract have been described. We are describing a case of a 79-year old woman with micropapillary urothelial carcinoma involving ureter and review the literature of this rare entity.

MAGE-A3/4 and NY-ESO-1 antigens expression in metastatic esophageal squamous cell carcinoma.

In the present study we analyzed immunohistochemical expression of MAGE-A 3/4 and NY-ESO-1 in 55 samples of esophageal squamous cell carcinomas (ESCC) and their respective lymph node metastases. To our knowledge this is the first study to assess and compare the expression of these antigens in ESCC lymph node metastases. Fifty (90.9%) primary ESCC were positive for MAGE-A 3/4 and 53 (96.6%) were positive for NY-ESO-1. MAGE-A 3/4 was expressed in all lymph node metastases and the intensity of expression was high in a majority of cases. NY-ESO-1 was negative in 2 (7.1%) lymph nodes metastases, while the reaction was predominantly moderate in the positive group. In primary tumors MAGE-A 3/4 showed a significantly higher intensity of expression compared to NY-ESO-1 (P=0.047), while in lymph node metastases the intensity of expression was not significantly different (P=0.387). Primary tumors with and without lymph node metastases showed no significant differences in MAGE-A 3/4 (P=0.672) and NY-ESO-1 (P=0.444) expression. Intensity of MAGE-A 3/4 (P=0.461) and NY-ESO-1 (P=0.414) expression in primary tumors was not significantly different compared to the expression in their respective lymph nodes metastases. Expression of MAGE-A 3/4 in primary tumors showed significant positive correlation with primary tumor expression of NY-ESO-1 (P=0.021) but no significant correlation with the expression of MAGE-A 3/4 in lymph node metastases (P=0.056). Expression of NY-ESO-1 in primary tumors showed significant positive correlation with the expression of NY-ESO-1 in lymph node metastases (P=0.001) and significant negative correlation with patients’ age (P<0.001). Expression of MAGE-A 3/4 and NY-ESO-1 in primary tumors and lymph node metastases showed no significant correlation with prognostic parameters such as tumor grade and TNM stage (P>0.05). We have shown different levels of MAGE-A 3/4 and NY-ESO-1 expression in almost all specimens of primary tumor and lymph node metastases, suggesting that ESCC may be possible target of immunotherapy and anti-tumor vaccination. High levels of expression in lymph node metastases indicate possible clinical benefit of postoperative vaccine with MAGE-A3 and NY-ESO-1 in advanced stage of disease.

Prognostic value of galectin-3 in primary cutaneous melanoma.

BACKGROUND: Galectin-3, one of the ß-galactoside-binding lectins, has been suggested as a marker of disease progression in melanoma patients because of its overexpression observed in recent studies. However, prognostic value of galectin-3 in primary cutaneous melanoma (PCM) has not been clearly defined. OBJECTIVES: The aim of the study was to analyse whether the intensity of galectin-3 expression can predict survival in patients with PMC. METHODS: Galectin-3 expression was evaluated using immunohistochemistry in 104 PCM samples, including 71 (68.2%) superficial spreading (SSM) and 33 (31.8%) nodular melanomas (NM). RESULTS: Significant difference of galectin-3 expression between SSM and NM was determined (P < 0.001). Increased galectin-3 expression was positively correlated with tumour thickness (P < 0.001), Clark (P < 0.001) and Breslow (P < 0.001) stage, mitotic rate (P < 0.001), presence of tumour ulceration (P < 0.001), lymphatic invasion (P = 0.018), positive sentinel lymph node (P < 0.022) and distant metastases (P < 0.001). Kaplan-Meier analysis showed an association between increased galectin-3 expression and reduced recurrence-free survival (RFS) (P = 0.001) and reduced disease-specific survival (DSS) (P = 0.015). In Cox proportional hazards regression analysis, significant predictors of reduced RFS were positive sentinel lymph node (P = 0.025) and lymphovascular invasion (P = 0.021), whereas predictors of DSS were tumour thickness (P = 0.012), lymphovascular invasion (P = 0.047), Clark stage (P = 0.029) and location of tumour on upper extremities (P = 0.024). CONCLUSIONS: Our results support the potential role of galectin-3 in PCM development, progression and metastasis. Moreover, galectin-3 could serve as an additional prognostic marker that might help in further stratifying the risk of disease progression and metastasis in patients with PMC.

Expression and prognostic role of syndecan-2 in prostate cancer.

Syndecans are a four-member family of transmembrane heparan sulphate proteoglycans that have different functions in cell signalling, adhesion, cytoskeleton organization, migration, proliferation, and angiogenesis. Several studies investigated the role of syndecan-2 (SDC2) in different carcinomas; however, only one being focused on SDC2 in prostate cancer. SDC2 expression and relationship with established prognostic features were assessed in a cohort of 86 patients treated with radical prostatectomy for clinically localized prostate adenocarcinoma. SDC2 expression was present in the majority of prostate cancers and absent in only 11.6% of cases. SDC2 expression was also recorded in cells of prostatic intraepithelial neoplasia, whereas normal prostatic epithelial tissue and stroma did not express SDC2. SDC2 overexpression in prostate cancer was significantly associated with established features indicative of worse prognosis such as higher preoperative PSA (P=0.011), higher Gleason score (P<0.001), positive surgical margins (P<0.003), and extraprostatic extension of disease (P<0.003). Moreover, expression of SDC2 was also associated with biochemical disease progression on univariate analysis (P<0.001). Study results supported the potential role of SDC2 in prostatic carcinogenesis and cancer progression. Moreover, SDC2 could serve as an additional prognostic marker that might help in further stratifying the risk of disease progression in patients with prostate cancer.

Myofibroblastic stromal reaction and expression of tenascin-C and laminin in prostate adenocarcinoma.

The aim of this study was to analyse relationship between changes of the stroma and expression of tenascin-C (TN-C) and laminin in prostate carcinoma. Tenascin-C immunostaining was increased, and laminin decreased in carcinomas compared with peritumoral tissue and benign prostate hyperplasia (P<0.05). Statistical analysis confirmed connection between stromal changes and TN-C expression in prostate carcinoma (P<0.05). Gleason pattern 3 carcinomas showed more pronounced stromal reaction and TN-C expression compared with Gleason pattern 4 carcinomas (P<0.05). The main cells in prostate cancer stroma are myofibroblasts that are also responsible for tenascin production. Degradation of laminin was not connected with myofibroblastic stromal changes.

Primary glomangioma of the esophagus mimicking esophageal papilloma.

We report a case of glomangioma of the esophagus in a 28-year-old woman who presented with a 3-year history of vague discomfort, pain and heat in the neck. At initial gross examination, the tumor mimicked an esophageal papilloma. The resected esophageal specimen contained a polypoid, whitish-gray mass, which measured 3 cm in maximum diameter. Microscopically the tumor consisted of loose fibrovascular stroma heavily infiltrated with mononuclear inflammatory cells and covered with focally hyperkeratotic, parakeratotic and acanthotic squamous epithelium without atypia. In the deeper area immediately above the true muscular layer of the esophageal wall, microscopical examination revealed the neoplasm consisting of numerous, small-to-medium branched vessels covered by regular endothelium and filled with erythrocytes. The loose stroma around the vessels contained poorly circumscribed nests of small, round to oval cells with a uniform appearance. Immunohistochemically, the tumor cells were immunoreactive for smooth muscle actin and vimentin and non-immunoreactive for CD34, CD117, desmin, pan-cytokeratin, synaptophisin, neuron-specific enolase and S-100 protein. Despite its bland histology, the infiltrative growth pattern was suggestive of aggressive behavior; thus, an appropriate clinical follow-up was recommended. An accurate diagnosis and an understanding of the behavior of these rare tumors, especially in an unusual location, are crucial to their management and clinical outcome.

Correlation between Bcl-2 and Bax in atrophic and hypertrophic type of actinic keratosis.

BACKGROUND: Recent investigations consider actinic keratosis (AK) as an earliest visible pattern of squamous cell carcinoma (SCC). We have analysed the expression of apoptosis-related proteins TP53, Bcl-2 and Bax in 30 atrophic and 30 hypertrophic AK cases. MATERIAL AND METHODS: Immunohistochemical analysis was performed following microwave streptavidin immunoperoxidase protocol on DAKO TechMate Horizon automated immunostainer (DAKO, Copenhagen, Denmark). Monoclonal antibody for TP53 and Bcl-2 and polyclonal antibody for Bax (DAKO, Copenhagen, Denmark) were used. RESULTS: Expression of TP53 showed no significant differences between two analysed groups (chi2-test, P = 0.35636) whereas expression of Bcl-2 and Bax protein was significantly higher in atrophic compared to hypertrophic AK (chi2-test, P = 0.01458 and P = 0.00358, respectively). Comparison of Bcl-2 : Bax ratio in two analysed AK showed significantly higher value in hypertrophic compared to atrophic AK (Mann-Whitney U test, P = 0.02272). Statistical analysis did not show any correlation between patient's sex and age, localization and size of the lesion with expression of investigated oncoproteins (anova, P > 0.05). CONCLUSIONS: Our results may indicate higher resistance of keratinocytes on apoptotic stimuli in hypertrophic compared to atrophic AK. Thus, we suppose that keratinocytes in hypertrophic AK live longer and probably have higher propensity for additional mutations and conversion to overt SCC.

Renal artery changes in patients with primary renal cell carcinoma.

Arterial fibromuscular dysplasia (FMD) is a noninflammatory, nonatherosclerotic, occlusive condition of the systemic arteries, most frequently affecting renal arteries. Renal cell carcinoma (RCC) might be associated with arterial hypertension; however, there are no data in the literature regarding the relationship between RCC and associated renal artery changes. We analyzed a consecutive series of 57 (35 male and 22 female) patients aging from 35 to 79 years (mean 58.9 years) who underwent nephrectomy due to RCC in the year 2003. The patients had RCC measuring from 2 to 16 cm (mean 7.1 cm). Specimens were routinely fixed, embedded in paraffin, cut, and stained with hematoxylin and eosin, Mallory trichrome method, and orcein. Renal arteries of 26 patients (20 male, 6 female) showed no changes. In these patients, RCC measured 2.5-11 cm in largest diameter (mean 6.6 cm). In 24 patients (10 male, 14 female), renal arteries showed FMD. RCCs in these patients measured between 2 and 16 cm (mean 8.0 cm). Seven patients had atherosclerotic changes in renal arteries. In this series, FMD was found in a significant proportion of patients with RCC, mainly in women. The cause of such changes and their relationship with RCC and systemic hypertension should be further analyzed.

Syringoid eccrine carcinoma.

A rare case of a syringoid eccrine carcinoma in a 52-year-old male patient is described. During the first hospitalization, syringoma was diagnosed, both clinically and histologically, keeping in mind the possibility of its malignant alteration. Difficulties in making a diagnosis with histological and immunohistochemical examinations are reported. The possibility of differential diagnosis of primary breast carcinoma and methods of its exclusion are also described. Treatment of the patient was surgical with good results.

Parachordoma: a case report.

Parachordoma is a very rare soft tissue tumor with histological features similar to chordoma and chondrosarcoma. It should be distinguished from metastatic chordoma and extraskeletal myxoid chondrosarcoma because of its different treatment and prognosis. In this paper we report one case of parachordoma in a 20-year-old female patient. The tumor occurred in the subcutaneous tissue of the left hand as a painless, fixed, slow-growing mass. Pathologic analysis revealed a tumor composed of lobules of cells with variably vacuolated cytoplasm (physaliphorous cells) separated by fibrous septa, predominantly arranged in peculiar small or large alveolar structures. Immunohistochemistry showed positive staining of the tumor cells with cytokeratin 8/18, S-100 protein and vimentin. The patient is well and without recurrence 20 months after surgery.

Langerhans' cell granulomatosis in an adult: a 22-year follow up.

A 57-year-old patient with Langerhans' cell granulomatosis (LCG) is presented. During 22 years of treatment, there were five relapsing infiltrations at different sites of the skull bones, which were treated by surgery, local radiotherapy, and chemotherapy. During the last relapse, the right temporal bone was infiltrated by granuloma and the petrous bone was destroyed with an intimate spread to the internal carotid artery. After two palliative surgical resections and ineffective radiotherapy, 12 cycles of chemotherapy (vinblastine plus prednisolone) were applied and a clinical remission of the disease was achieved. Special attention is paid to the complexity of diagnosis and choice of therapy. It is concluded that the behavior of LCG may change with time, and assume an aggressive form of the disease. Chemotherapy is the treatment of choice for this type of multifocal malignant form of LCG. Prognosis of the disease is unpredictable.

nNOS expression in reactive astrocytes correlates with increased cell death related DNA damage in the hippocampus and entorhinal cortex in Alzheimer's disease.

The immunocytochemical distribution of the neuronal form of nitric oxide synthase (nNOS) was compared with neuropathological changes and with cell death related DNA damage (as revealed by in situ end labeling, ISEL) in the hippocampal formation and entorhinal cortex of 12 age-matched control subjects and 12 Alzheimer's disease (AD) patients. Unlike controls, numerous nNOS-positive reactive astrocytes were found in AD patients around beta-amyloid plaques in CA1 and subiculum and at the places of clear and overt neuron loss, particularly in the entorhinal cortex layer II and CA4. This is the first evidence of nNOS-like immunoreactivity in reactive astrocytes in AD. In contrast to controls, in all but one AD subject, large numbers of ISEL-positive neuronal nuclei and microglial cells were found in the CA1 and CA4 regions and subiculum. Semiquantitative analysis showed that neuronal DNA fragmentation in AD match with the distribution of nNOS-expressing reactive astroglial cells in CA1 (r = 0.74, P < 0.01) and CA4 (r = 0.58, P < 0.05). A portion of the nNOS-positive CA2/CA3 pyramidal neurons was found to be spared even in the most affected hippocampi. A significant inverse correlation between nNOS expression and immunoreactivity to abnormally phosphorylated tau proteins (as revealed by AT8 monoclonal antibody) in perikarya of these CA2/3 neurons (r = -0.85, P < 0.01) suggests that nNOS expression may provide selective resistance to neuronal degeneration in AD. In conclusion, our results imply that an upregulated production of NO by reactive astrocytes may play a key role in the pathogenesis of AD.

DNA ploidy analysis and cell proliferation in congenital sacrococcygeal teratomas.

BACKGROUND: Congenital sacrococcygeal teratoma is the most common germ cell tumor in infants and children. It usually is diagnosed at birth, is benign, and consists of fully differentiated mature tissues. Congenital sacrococcygeal teratomas (SCTs) also may contain immature tissues, most commonly of neural origin. The proportion of malignant teratomas increases with advancing age, but the relation between mature and immature SCTs is not well understood. Thus, it is very important to determine proliferative activity, DNA ploidy, and DNA index to predict biologic behavior of these tumors. METHODS: DNA ploidy and cell proliferation were analyzed by flow cytometry, and the expression of proliferating cell nuclear antigen (PCNA) and Ki-67 were analyzed immunohistochemically on paraffin embedded tissue. RESULTS: All the tumors that were surgically treated within 3 months after birth, including immature teratoma, were diploid. Strongly positive PCNA immunostaining was found in both immature teratomas, and weakly positive PCNA was found in nine cases. Weak positivity for Ki-67 was observed in 2 cases, and moderate positivity was observed in 6 cases including immature teratomas. CONCLUSION: The value of flow cytometry in the prediction of biologic behavior of congenital SCT should be analyzed further. Our results suggest that Ki-67 and especially PCNA may reflect the proliferative activity of these tumors.

Ultrastructural analysis and TUNEL demonstrate motor neuron apoptosis in Werdnig-Hoffmann disease.

Werdnig-Hoffmann disease (WHD) is the most severe clinical type of spinal muscular atrophy characterized by loss of lower motor neurons and paralysis. We examined the hypothesis that disease pathogenesis is based on an inappropriate persistence of normally occurring motor neuron programmed cell death. The diagnosis of WHD was made on the basis of clinical findings, electromyoneurography, and biopsy, and further confirmed by mutation analysis of the survival motor neuron (SMN) and neuronal apoptosis inhibitory protein (NAIP) genes using PCR. We used ultrastructural analysis as well as TUNEL and ISEL methods to assess DNA fragmentation, and immunocytochemistry to identify expression of the apoptosis-related proteins bcl-2 and p53. A significant number of motor neurons in the spinal cord of children with WHD were shown to die by apoptosis. As revealed by TUNEL, dying neurons in WHD patients comprised 0.2%-6.4% of the neuron numbers counted. This finding contradicts earlier studies that failed to find such evidence and suggests that early blockade of prolonged motor neuron apoptosis may be a potential therapeutic strategy for WHD.

Calcified leiomyoma of the lateral pterygoid muscle in an 8-year-old boy.

Deep soft tissue leiomyomas are extremely rare benign tumors in childhood. An unusual case of benign calcified leiomyoma of the pterygoid muscle in an 8-year old boy is presented. Clinical manifestations and radiographic and histologicpathologic findings, as well as the differential diagnosis and possible histogenesis of this rare tumor, are discussed.

[Carcinoma of the urinary bladder resembling lymphoepithelioma]. / Karcinom mokracnog mjehura slican limfoepiteliomu.

Lymphoepithelioma-like carcinoma of the bladder is rare and has only recently been described at this site. We report a case of lymphoepithelioma-like carcinoma of the urinary bladder in a 70-years old male patient who presented with painless hematuria lasting for one month. The patient underwent transurethral resection two times. Histopathologic diagnosis was transitional cell carcinoma. Histopathologic examination of the third biopsy showed tumor tissue with typical syncitial growth pattern of atypical, large, epitheloid cells with ill-defined cytoplasmic borders, prominent nucleoli and numerous mitoses. A prominent lymphocytic infiltrate was found as a component of the tumor. Immunohistochemistry showed positive reaction for cytokeratin and epithelial membrane antigen, and negative results for leukocyte common antigen, CD3, CD20, CD30, CD68 and PSA. The lymphoid infiltrate was an admixture of T and B cells. The tumor invaded the muscle wall. After last surgery the patient underwent chemotherapy by local application of Adriablastine. At present the patient is without recurrence 10 months after the last surgery.

Inflammatory myofibroblastic tumor with extensive involvement of the bowel in a 7-year-Old child.

We present a case of unusual localization of inflammatory fibroblastic tumor in the terminal ileum, cecum, and ascending colon in a 7-year-old child. Segmental resection of the terminal ileum, cecum, and ascending colon with a tumor mass up to 6 cm in diameter was performed. Pathohistological examination of biopsy specimen was performed on routine hematoxylin-eosin sections, as well as immunohistochemically with primary antibodies to CD3, CD20, CD68, factor VIII, vimentin, smooth muscle actin, desmin, cytokeratin and S-100 protein, and k and l light chains. The tumor was composed of highly vascularized tissue with interlacing fascicles of elongated spindle cells admixed with plasma cells, histiocytes, lymphocytes, and eosinophils. The diagnosis of inflammatory myofibroblastic tumor was confirmed by immunohistochemistry. Inflammatory myofibroblastic tumor cannot be distinguished clinically from highly malignant neoplasm or some other conditions. Surgical resection and careful pathohistological analysis are needed, and a long-term follow-up is recommended.