CYP24A1 expression inversely correlates with melanoma progression: clinic-pathological studies.

The major role of 24-hydroxylase (CYP24A1) is to maintain 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) homeostasis. Recently, it has been discovered that CYP24A1 also catalyses the hydroxylation of 20(OH)D3, producing dihydroxy-derivatives that show very effective antitumorigenic activities. Previously we showed a negative correlation of vitamin D receptor (VDR) and CYP27B1 expression with progression, aggressiveness and overall or disease-free survivals of skin melanomas. Therefore, we analyzed CYP24A1 expression in relation to clinicopathomorphological features of nevi, skin melanomas and metastases. In melanocytic tumors, the level of CYP24A1 was higher than in the normal epidermis. The statistically highest mean CYP24A1 level was found in nevi and early stage melanomas. With melanoma progression, CYP24A1 levels decreased and in advanced stages were comparable to the normal epidermis and metastases. Furthermore, the CYP24A1 expression positively correlated with VDR and CYP27B1, and negatively correlated with mitotic activity. Lower CYP24A1 levels correlated with the presence of ulceration, necrosis, nodular type and amelanotic phenotypes. Moreover, a lack of detectable CYP24A1 expression was related to shorter overall and disease-free survival. In conclusion, the local vitamin D endocrine system affects melanoma behavior and an elevated level of CYP24A1 appears to have an important impact on the formation of melanocytic nevi and melanomagenesis, or progression, at early stages of tumor development.

The analysis of receptor-binding cancer antigen expressed on SiSo cells (RCAS1) immunoreactivity within the microenvironment of the ovarian cancer lesion relative to the applied therapeutic strategy.

RCAS1 is involved in generating the suppressive profile of the tumor microenvironment that helps cancer cells evade immune surveillance. The status of the cells surrounding the cancer nest may affect both the progression of the cancer and the development of metastases. In cases of ovarian cancer, a large number of patients do not respond to the applied therapy. The patient's response to the applied therapy is directly linked to the status of the tumor microenvironment and the intensity of its suppressive profile. We analyzed the immunoreactivity of RCAS1 on the cells present in the ovarian cancer microenvironment in patients with the disease; these cells included macrophages and carcinoma-associated fibroblasts. Later we analyzed the immunoreactivity levels within these cells, taking into consideration the clinical stage of the cancer and the therapeutic strategy applied, such as the number of chemotherapy regiments, primary cytoreductive surgery, or the presence of advanced ascites. In the patients who did not respond to the therapy we observed significantly higher immunoreactivity levels of RCAS1 within the cancer nest than in those patients who did respond; moreover, in the non-responsive patients we found RCAS1 within both macrophages and carcinoma-associated fibroblasts. RCAS1 staining may provide information about the intensity of the immuno-suppressive microenvironment profile found in cases of ovarian cancer and its intensity may directly relate to the clinical outcome of the disease.

[Evaluation of the number of enterochromaffin cells in gastric mucosa in subjects with functional dyspepsia]. / Ocena liczby komórek enterochromafinowych w blonie sluzowej zoladka u osób z dyspepsja czynnosciowa.

UNLABELLED: Enterochromaffin cells (EC) in gastric mucosa produce of serotonin and melatonin and trough its paracrine activity able to change motoric and secretory function. AIM OF OUR STUDIES: To evaluate the density of enterochromaffin cells in gastric mucosa in subjects with functional dyspepsia. MATERIAL AND METHODS: The investigations were performed in 50 subjects including 25 subjects with Postprandial Distress Syndrome and 25 subjects with Epigastric Pain Syndrome accordingly to Rome Criteria III. The comparative group comprised 25 healthy subjects. To identify enterochromaffin cells the bioptates were collected from the corpus and prepyloric part of the stomach and stained immunohistochemically. RESULTS: Number of EC cells in corpus part were--healthy subjects 1.68 +/- 0.35, in patients with PDS--1.26 +/-0.37 (p < 0.001) and in EPS--2.11 +/- 0.43 (p < 0.01). In antrum Number of EC cells were: K--2.52 +/- 0.52, PDS--1.79 +/- 0.3 (p < 0.01), EPS--2.69 +/- 0.46 (p > 0.05). The results of urea breath test (UBT-13C) were: PDS--13.10 +/- 6.49% per hundred, EPS--18.98 +/- 10.11% per hundred (p < 0.05). In group of patients with EPS the positive correlation between number of enterochromaffin cells and urea breath test results was observed. CONCLUSIONS: The number of EC cells in gastric mucosa in subjects with functional dyspepsia is different than in healthy subjects. Evaluation of the number of enterochromaffin cells could have clinical value in differential diagnosis in functional dyspepsia.

Decreased expression of CYP27B1 correlates with the increased aggressiveness of ovarian carcinomas.

CYP27B1 hydroxylates 25-hydroxyvitamin D3 in position C1α into biologically active 1,25-dihydroxyvitamin D3, calcitriol. CYP27B1 is expressed in normal tissues and tumors. Since calcitriol indicates anticancer activities and CYP27B1 expression can be deregulated during malignant progression, we analyzed its expression in ovarian cancers in relation to pathomorphological features of tumors and overall survival (OS). Expression of CYP27B1 was evaluated in 61 ovarian tumors, 18 metastases and 10 normal ovaries. Normal ovarian epithelium showed the highest levels CYP27B1 with a significant decrease in its expression in ovarian cancers. Both poorly differentiated primary tumors and metastases showed the lowest level of CYP27B1 expression, while non-metastasizing tumors showed a higher CYP27B1 level than tumors that developed metastases. The expression of CYP27B1 was positively correlated with a lower proliferation rate, lower dynamism of tumor growth and tumor infiltrating lymphocyte response. Furthermore, CYP27B1 expression was negatively correlated with tumor cell modeling of their microenvironment. CYP27B1 expression was also associated with longer OS time. In summary, our results suggest that local expression of CYP27B1 in ovarian tumor cells can modify their behavior and promote a less aggressive phenotype by affecting local concentrations of active of vitamin D levels within the tumor microenvironment.

[Pleural mesothelioma--case with effusion in both pleural cavities]. / Miedzybloniak oplucnej--przypadek z wysiekiem w obu jamach oplucnej.

Etiology of the pleural exudate is not always easy to establish with the routine diagnostic procedures. We report the history of a 55-year-old man, driver--without evident occupational exposure to asbestos dust. Patient was treated in hospital because of recurrent bilateral sanguineous pleural fluid. Repeated basic laboratory examinations of pleural exudate did not contribute to establishing etiology of the disease. At the beginning of hospital stay antituberculosis therapy was applied but was unsuccessful. Rapid accumulation of the fluid, deterioration of general condition of the patient, presence of dysplasia in the cells of the sediment of the exudate indicated possibility of diagnosis of neoplasm of the pleura. Intravenous injections of cisplatin and intrapleural application of bleomycin did not influence, however, the course of the disease. Final diagnosis was possible only after pleural biopsy (with Abrams needle) was performed. Histopathologic examination of the specimen disclosed: malignant mesothelioma biphasic type. Patient died after 3 months of observations and attempt at treatment.

[Histoclinic of nonspecific inflammatory bowel diseases]. / Histoklinika nieswoistych stanów zapalnych jelita grubego.

In the paper pathomorphological indices of changes in inflammatory bowel lesions were presented. Additionally a clinicopathological correlation has been performed. The necessity of collaboration between gastroenterologist and pathomorphologist in diagnostic process of inflammatory bowel lesions has been indicated. The collaboration should mainly result from the lack of repeated morphological changes in most cases of inflammatory bowel lesions, but also from lack of specificity of signs and symptoms in these inflammations.

Application of PCR methods to evaluate EGFR, KRAS and BRAF mutations in a small number of tumor cells in cytological material from lung cancer patients.

The epidermal growth factor receptor (EGFR) mutation status in the tyrosine kinase domain is known to be a predictor of the response to gefitinib or erlotinib in lung cancer; thus, a non-surgical procedure of tumor specimen collection is critical for mutation analysis. The aim of the present study was to analyze the EGFR, KRAS and BRAF status in limited cytological material. To the best of our knowledge, this is the first time that the quantitative scale of tumor cells and the percentage of tumor cells in cytological material were evaluated at the early stages of pathomorphological material qualification for EGFR, KRAS and BRAF mutation analysis. Our results revealed that even 100-1,000 tumor cells from fine needle aspiration (FNA) samples provided reliable results of mutation analysis when sensitive real-time polymerase chain reaction (PCR) methods were used. EGFR mutations were detected in 10% (7/71) and KRAS mutations were detected in 35% (19/54) of the lung adenocarcinoma cases. In addition, we reported the most common inhibiting mutation (p.T790M) found in coexistence with p.L858R in an FNA sample from a patient, for whom short-term improvement after erlotinib treatment was observed before further progression of the disease. Subsequently, mutual exclusion of EGFR and KRAS mutations was observed. Cytological samples with a small number of tumor cells obtained via FNA, endobronchial ultrasound (EBUS)-transbronchial needle aspiration (TBNA) or brushing are suggested to be used for diagnostic purposes after careful selection by cytopathologists and analysis using a validated, sensitive real-time PCR method.

Automated recognition and counting of the immunoreactive neuroendocrine cells in chronic gastritis (the preliminary study).

The paper presents the designed software CAMI (Computerized Analysis of Microscopic Images) for a digital reconstruction of the diversiform glands seen in chronic inflammatory gastric mucosa, and for automated recognition and quantization of the immunoreactive neuroendocrine (NE) cells appearing within mucosal glands. Digital reconstruction of the individual gastric gland is difficult due to variable shapes of the glandular cross-sections. Fifteen gastric biopsy specimens representing chronic gastritis were stained routinely with H+E and immunohistochemically with 3 NE markers: Chromogranin A, Somatostatin and Serotonin. Two expert pathologists counted manually the NE cells with the light microscope in 4 types of glandular cross-sections: round, short- oblique, long- oblique and longitudinal. The automated counting of the NE cells was performed on the digital images presenting the same microscopic areas which were selected for the manual reading. The first step of image analysis was concerned to the cell extraction and recognition of the cytoplasmic immunoreactivity. The unstained nuclei of the NE cells were spotted by the sequential thresholding algorithm combined with the artificial neural network of Support\Vector Machine (SVM) type. The second step of image analysis comprised reconstruction of the glands. The presumed shape of each gastric gland was defined by the cellular lining of viewed glandular cross-section. The designed algorithm for gland reconstruction was based on the cell masks. The third step of analysis dealt the cell counting. Every recognized gland with the face cells was used for the NE cell evaluation. The results of the automated quantization compared with manual counting results for the number of NE cells showed high concordance in 3 types of glandular cross-sections: round, short- and long- oblique. A difference noticed in the results of the longitudinal glands should be verified in the extended study. The designed software CAMI is more adequate for the gland recognition with an discontinuous gland face seen in the immunohistochemical digital images, which appear to be a difficult problem for the accurate automated analysis of the cellular component of glands.

Increased cyclooxygenase expression and thymine dimer formation after repeated exposures of humans to low doses of solar simulated radiation.

The impact of repeated doses of solar simulated radiation (SSR) has not been evaluated, particularly to determine if photoadaptation and photoprotection develop over time. In this study, erythema, pigmentation, cyclooxygenase (COX)-1 and 2 expression and thymine dimer (dTT) formation were evaluated in the skin of irradiated subjects of phototype II or III. Groups of 7-10 volunteers were whole-body irradiated with a low dose of SSR on each of 10 consecutive days followed by a single erythemal ultraviolet B (UVB) dose on a small body area, or irradiated only with the single erythemal UVB dose on a small body area, or irradiated with the low dose of SSR on each of 30 consecutive days, or were unirradiated. Erythema and pigmentation were measured 24 h after the final SSR or UVB, and skin biopsies collected for the assessment of COX(+) cells and dTT(+) nuclei. The repeated SSR exposures induced a small increase in pigmentation without erythema, and were slightly protective against the erythemal effects of the subsequent high UVB dose. The number of COX-1(+) and 2(+) cells increased as a result of 10-days SSR and rose still further after 30-days SSR, indicating that photoadaptation had not developed. The SSR exposures did not result in any protection against the further increase in COX-1 and 2 expression caused by the erythemal UVB dose. In contrast, for dTT formation, the repeated SSR exposures led to a limited degree of both photoadaptation and photoprotection.