Olga Stasikowska-Kanicka

Expression of arachidonate metabolism enzymes and receptors in nasal polyps of aspirin-hypersensitive asthmatics.

Background: The pathogenesis of rhinosinusitis in aspirin-exacerbated airway disease is closely linked to the disequilibrium in arachidonic acid metabolism. Although considerable amounts of data concerning impaired eicosanoid production are available, the precise mechanism and pathogenesis of the disease are still unknown. The aim of the present study was to assess the expression of enzymes belonging to the arachidonic acid cascade and receptors for arachidonate derivative metabolites in nasal polyps from aspirin- hypersensitive (AH) and aspirin-tolerant (AT) patients with rhinosinusitis. Methods: Cells expressing cysteinyl leukotriene (CysLT) receptors (CysLT1 and CysLT2), arachidonate 5-lipoxygenase, leukotriene B4 receptor type 1, E-prostanoid receptors (EP2 and EP4), cyclooxygenase (COX)-1 and COX-2 were detected by immunocytochemistry in nasal polyps obtained from 10 AH patients and 18 AT patients. Results: There was a significantly higher density of cells expressing CysLT1 and CysLT2 receptors in nasal polyps from AH patients than from AT patients (p < 0.001). In contrast, the density of cells expressing EP2 receptor and COX-2 was significantly lower in AH patients than in AT patients (p < 0.02). The number of COX-2-positive epithelial cells was significantly reduced in AH polyps (p < 0.04). Conclusions: The elevated number of nasal polyp cells expressing CysLT receptors and lack of cells expressing EP2 receptor and COX-2 may be related to a more severe course of hyperplastic rhinosinusitis in aspirin hypersensitivity.

Increased tissue immunoexpression of YKL-40 protein in high grade serous ovarian cancers

YKL-40 is a glycoprotein secreted by numerous human cells, such as cartilage, synovial, and endothelial cells. The biological role of YKL-40 has not yet been fully unveiled, however, its participation is perceived in angiogenesis, growth, proliferation, differentiation, and remodeling processes. The primary goal of our study was to evaluate possible differences in tissue immunoexpression of YKL-40, assumed between high grade and low-grade ovarian cancers and between the above-mentioned cancer types and benign lesions. Another purpose was to find out whether immunoexpression of the studied protein could correlate with the tumor proliferation process, evaluated by Ki-67 immunoexpression. The analysis comprised 45 women, diagnosed and treated for epithelial ovarian tumors at the Medical University of Lodz between 1997 and 2002. YKL-40 protein immunoexpression was semiquantitatively assessed, whereas immunoexpression of Ki-67 was evaluated using a computer image analysis system. Significantly higher immunoexpression values of both examined proteins were observed in high-grade serous ovarian cancers vs. low-grade and benign tumors. Moreover, a significant positive correlation was identified between the immunoexpressions of YKL-40 and Ki-67 proteins in the studied groups of tumors. In conclusion, the obtained data suggest an overt prominence of TKL-40 tissue immunoexpression of YKL-40 in high-grade serous ovarian tumors, which could then be approached as a helpful, additional marker to identify more aggressive ovarian cancers.

IL-17 Expression in Dermatitis Herpetiformis and Bullous Pemphigoid

Dermatitis herpetiformis (DH) and bullous pemphigoid (BP) are skin diseases associated with eosinophilic and neutrophilic infiltrations. Although cytokines are critical for the inflammatory process, there are single findings concerning concentration of IL-17 in bullous diseases. The goal of this study was to assess IL-17 expression in DH and BP patients. Skin biopsies were taken from 10 DH, 14 BP patients and from 10 healthy subjects. The localization and expression of IL-17 was studied by immunohistochemistry and the serum concentration was measured by immunoassays. Expression of IL-17 in the epidermis and in influxed cells in dermis was detected in skin biopsies. Expression of IL-17 was statistically higher in epidermis and infiltration cells in specimens from BP than from DH patients. Examined interleukin expression was detected in perilesional skin of all patients but it was much lower than in lesional skin. The expression of IL-17 was not observed in biopsies from healthy people. Serum level of IL-17 was statistically higher in BP and DH groups as compared to control group. Our results provide the evidence that IL-17 may play an essential role in activating and recruiting eosinophils and neutrophils, which ultimately contribute to the tissue damage in DH and BP.

Assessment of apoptosis and MMP-1, MMP-3 and TIMP-2 expression in tibial hyaline cartilage after viable medial meniscus transplantation in the rabbit.

Introduction The porpuse of this animal study was to assess chondrocyte apoptosis and MMP-1, MMP-3 and TIMP-2 expression in rabbit tibial cartilage 6 months after viable medial meniscal autografts and allografts. Material and methods Twenty white male New Zealand rabbits were chosen for the study. The medial meniscus was excised from 14 animals and stored under tissue culture conditions for 2 weeks, following which t of them were implantated as autografts and 7 as allografts. The control group consisted of 6 animals which underwent arthtrotomy. When the animals were eutanized, the tibial cartilage was used for immunohisochemical examination. Apoptosis (TUNEL method) and MMP-1, MMP-3 and TIMP-2 expression were estimated semiquantatively. Results An increased level of chodrocyte apoptosis in the tibail cartilage was observed after both kinds of transplants (p < 0.05), allografts (1.43 ±0.98) and autografts (0.86 ±0.69); no statistical diferences existed between them. An increased level of metalloproteinases and TIMP-2 expression was obreved only after allografts with statistical differences among the allograft group, the autograft group nad the control group (p < 0.05). Conclusions Our findings suggest that the meniscal graft does not protect the hyaline cartilage against excessive apoptosis. The results of experimantal studies on humans indicate the need to device a method of apoptosis inhibition in the hyaline cartilage to improve long-term results of meniscal transplantation.

Effect of human papillomavirus on cell cycle-related proteins p16INK4A, p21waf1/cip1, p53 and cyclin D1 in sinonasal inverted papilloma and laryngeal carcinoma. An in situ hybridization study

Human papillomavirus (HPV) infection is implicated as an important risk factor in the development of head and neck cancers. Many studies focusing on the relationships between HPV infection and cell cycle proteins immunoexpression in laryngeal lesions have provided contradictory results. The aim of this study was to evaluate the relationships between HPV DNA presence and p16INK4a, p21waf1/cip1, p53 and cyclin D1 immunoexpression in heterogenous HPV-positive and HPV-negative groups of laryngeal cancers and inverted papillomas. The HPV DNA expression was detected using an in situ hybridization method and immunoexpression of p16INK4a, p21waf1/cip1, p53 and cyclin D1 using immunohistochemistry. The immunoexpression of p21waf1/ /cip1 and p53 proteins was lower in the HPV-positive group compared to the HPV-negative group, although only the difference of p53 staining was statistically significant. The immunoexpression of p16INK4a and cyclin D1 was significantly increased in the HPV-positive group compared to the HPV-negative group. The increased immunoexpression of p16INK4a and cyclin D1, and the lower immunoexpression of p21waf1/cip1 and p53 in the HPV-positive group compared to the HPV-negative group, supports the hypothesis that HPV may play an important role in cell cycle dysregulation.

Mediators of Mast Cells in Bullous Pemphigoid and Dermatitis Herpetiformis

Bullous pemphigoid (BP) and dermatitis herpetiformis (DH) are skin diseases associated with inflammation. However, few findings exist concerning the role of mast cells in autoimmune blistering disease. Skin biopsies were taken from 27 BP and 14 DH patients, as well as 20 healthy individuals. Immunohistochemistry was used to identify the localization and mast cell expression of TNFα and MMP9 in skin lesions and perilesional skin. The serum concentrations of TNFα, MMP9, chymase, tryptase, PAF, and IL-4 were measured by immunoassay. TNFα and MMP9 expression in the epidermis and in inflammatory influxed cells in the dermis was detected in skin biopsies from patients. Although these mediators were found to be expressed in the perilesional skin of all patients, the level was much lower than that in lesional skin. Increased serum PAF levels were observed in BP patients. Mast cells may play an essential role in activating inflammation, which ultimately contributes to the tissue damage observed in BP and DH. Our findings suggest that differences in the pattern of cytokine expression directly contribute to variations in cellular infiltration in DH and BP.

Immunohistochemical study on survivin in sinonasal tumors and its relationship with the immunoexpression of Ki67 and Bcl-2

The immunoexpression of the inhibitor of apoptosis protein survivin has been shown to be a significant prognostic factor in various human cancers. Immunohistochemical method was used to examine the expression of survivin, Ki67 and Bcl-2 in 20 cases of sinonasal inverted papillomas (IPs), 12 cases of sinonasal squamous cell carcinoma (SNCs) and 19 cases of nasal chronic sinusitis as a control. Nuclear immunostaining for survivin was observed in 14 of 20 (70%) cases of sinonasal IPs and 10 of 12 (83.4%) cases of SNCs. Apart from nuclear, also weak cytoplasmic immunoexpression of survivin was detected in 2 of 20 cases (10%) of sinonasal IP and moderate intense staining in 9 of 12 cases (75%) of SNC. There was no immunostaining for survivin in 19 control cases. The immunoexpression of survivin, Ki67 and Bcl-2 was significantly higher in SNCs than in sinonasal IPs and control group. Moreover, nuclear survivin and Ki67 antigen immunoexpression were significantly higher in sinonasal IPs group as compared to control group. There were statistically significant positive correlations between nuclear (but not cytoplasmic) immunoexpression of survivin and Ki67 antigen, as well as Bcl-2 oncoprotein in both tested tumors. In conclusion, our findings suggest that survivin, Ki67 and Bcl-2 may be involved in sinonasal tumorigenesis.

Evidence for apoptosis, MMP-1, MMP-3 and TIMP-2 expression and their effect on the mechanical and biochemical properties of fresh viable knee medial meniscal allografts and autografts in the rabbit

Introduction The study sought evidence for apoptosis, the expression of MMP-1, MMP-3 and TIMP-2 and their effect on the mechanical and biochemical properties of rabbit fresh knee medial meniscal grafts in a 6-month follow-up. Material and methods Forty white male New Zealand rabbits were chosen for the study. The medial meniscus was excised from 28 animals and stored under tissue culture conditions for 2 weeks, following which 14 of them were implanted as autografts and 14 as allografts. When the animals were euthanized, 20 menisci were used for immunohistochemical examinations. Apoptosis (TUNEL method) and MMP-1, MMP-3 and TIMP-2 immunoexpression were estimated semiquantitatively. The other 20 menisci were subjected to biochemical analysis and their degree of elasticity was evaluated. Results An increased level of apoptosis (p <0.05) was observed both in allografts (1.57 ±0.98) and autografts (0.86 ±0.69); no statistical differences existed between them. An increased level of metalloproteinases and TIMP-2 expression was observed only in the allografts (p < 0.05). The highest decrease of degree of elasticity and the most significant changes in biochemical composition were observed in allografts (p < 0.05). Conclusions The studies confirmed the existence of excessive apoptosis in both kinds of fresh viable medial meniscal implants: auto- and allografts. Our results suggest that apoptosis and increased MMP-1 and MMP-3 expression have an adverse effect on the biological properties of implants. The results of experimental studies on humans indicate the need to devise a method of apoptosis inhibition in transplanted menisci to improve long-term results.

Opioid-receptor gene expression and localization in cancer cells

This study examined the presence and cellular localization of three types of opioid receptors (MOR, DOR and KOR) in five human cancer cell lines: MCF-7, MDA-MB-231, HT-29, MGH-U1 and SH-SY5Y. Expression levels of opioid receptors were measured quantitatively using real-time PCR, and the localizations of the receptors in the cells were determined by immunocytochemistry. All three types of opioid receptors were present in each of the five cell lines examined. However, three of the cell lines (MCF-7, HT-29 and SH-SY5Y) showed significantly higher levels of MOR mRNA and protein than the other two types of receptors (DOR and KOR). Immunocytochemistry revealed that MOR, DOR and KOR receptors were predominantly present on the surface of cell membranes, although these receptors were also occasionally present in the cell cytoplasm.

E-cadherin expression is more associated with histopathological type of thyroid cancer than with the metastatic potential of tumors

The aim of this study was to evaluate the relationship between abnormal expression of E-cadherin (E-CAD) and the extracapsular extension of tumors, lymph node involvement and the presence of metastasis in various types of thyroid cancers. Histopathological specimens of 35 benign thyroid lesions and 122 malignant tumors (papillary, follicular, poorly differentiated and undifferentiated cancers) were analyzed. E-CAD immunostaining intensity, its subcellular localization, homogeneity within lesion, and the relation of staining intensity between tumor and surrounding thyroid parenchyma were evaluated. The obtained results show that the variants of differentiated cancers with a poorer prognosis (i.e. tall cell and follicular variants of papillary cancer and widely invasive follicular cancers) present reduced intensity of E-CAD expression, its abnormal localization or heterogeneity of staining more frequently than classical papillary cancers and minimally invasive follicular cancers. However, the assessment of E-CAD expression does not allow the prediction of extrathyroidal growth of thyroid cancers.