Małgorzata Wągrowska-Danilewicz

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.

Trends in the Incidence of Biopsy-Proven Glomerular Diseases in the Adult Population in Central Poland in the Years 1990-2010

Background: There is a paucity of epidemiological data on biopsy-proven glomerulonephritis (GN) in Poland. The aim of this study was to assess the epidemiology of renal diseases based on histological diagnosis in the region of Central Poland over the last two decades. Methods: Retrospective analysis of the results of 746 consecutive native kidney biopsies performed in the Caucasian adults from 1990 to 2010 in a single tertiary nephrology center serving an area of Central Poland. Results: Primary GN was found in 81.4% of all biopsies. The mesangioproliferative GN including IgA nephropathy was the most frequent type of primary GN (51.2%). Membranoproliferative GN was diagnosed between 1990 and 2000 more frequently than in the following decade (26.7 vs. 7.3%, p < 0.001). There was a significant increase in the incidence of FSGS (4.8 vs. 17.3%, p < 0.001) and MCD (5.1 vs. 11.2%, p < 0.001) over time. Secondary GN was documented in 18.6% of biopsies and lupus nephritis was the most frequent cause (34.5%). Conclusion: Our analysis showed the decrease in the incidence of membranoproliferative GN with the parallel increase of FSGS and MCD over the last twenty years. Mesangioproliferative GN including IgA nephropathy remains the most frequent type of GN observed in our region.

Amyloidosis of the head and neck: a clinicopathological study of cases with long-term follow-up

Extracellular deposits of insoluble proteinaceous material giving a starch-like reaction when treated with iodine and sulphuric acid and accumulating in tissue was for the first time described by Rokitansky in 1842 [1]. It was not until 1851 that Virchow applied the term “amyloidosis” to describe this deposition [2]. An amorphous substance called amyloid (insoluble fibril-forming protein) is deposited in extracellular spaces of organs and tissues. Amyloid deposits, under the electron microscope, appear as non-branching fibrils with a cross-linked, β-pleated sheet conformation. They are eosinophilic after haematoxylin–eosin staining and display apple-green birefringence with polarized light when stained with Congo red. Chronic inflammations of bacterial or non-bacterial origin and immunological immune-competent neoplasm cells are examples of factors that induce amyloid fibril biosynthesis. The organs and tissues where amyloid deposits occur become stiff and plastic, having a hyaline-like appearance that leads to a loss of previous function [3].

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.

Expression of α5β1 and α6β1 integrins in IgA nephropathy (IgAN) with mild and severe proteinuria. An immunohistochemical study

Immunoperoxidase staining was carried out using monoclonal antibodies against integrin α5β1 and integrin α6β1 on renal biopsy specimens from patients with IgA nephropathy with mild proteinuria (n = 15) and with severe proteinuria (n = 10). Our study revealed increase in glomerular immunoexpression of α5β1 in renal biopsies in IgA nephropathy with severe proteinuria. There were no statistical differences between interstitial α5β1 immunostaining, as well astubular α6β1 immunoexpression in renal tissue between patients with mild and severe proteinuria. The intensity of interstitial α5β1 integrin immunoexpression was positively correlated with the degree of interstitial fibrosis in both studied groups, meanwhile the intensity of tubular α6β1 integrin immunoexpression was not related to the degree of renal interstitial fibrosis in patients with mild and severe proteinuria. Our results suggest that elevated immunoexpression of α5β1 integrin on endothelial glomerular cells in the renal tissue in patients with severe proteinuria may point to the role of this integrin in the mechanism of glomerular injury in these cases of IgA nephropathy. The positive association between the interstitial expression of α5β1 integrin and the relative interstitial cortical volume in renal biopsies in patients with mild and severe proteinuria suggests that α5β1 integrin may play a role in the pathogenesis of chronic progressive renal disease in both studied group. Lack of positive correlation between tubular α6β1 integrin immunoexpression and the relative interstitial cortical volumen may indicate that this molecule play no role in the patomechanism of interstitial fibrosis in IgA nephropathy with mild and severe proteinuria.

Quantitative Analysis of Interstitial Mast Cells in AA and AL Renal Amyloidosis

Eighteen renal biopsy specimens obtained from patients with AA-type renal amyloidosis (AA) and 11 from patients with AL-type renal amyloidosis (AL), for whom both light and electron microscopy as well as immunofluorescence microscopy and full clinical data were available, were examined quantitatively. The cases were selected on the basis of immunohistochemical studies. As a control, we used 10 biopsy specimens from the kidneys removed because of trauma. Morphometric investigations were carried out by a computer image analysis system to find an answer to the question of whether mast cells can correlate with tubulointerstitial fibrosis in AA and AL renal amyloidosis, and to examine the relationship between mast cells and interstitial alpha-smooth muscle actin (alpha-SMA) expression and interstitial infiltrates. The morphometric study revealed that the mean values of the interstitial tryptase-positive cells, expression of alpha-SMA, interstitial volume, CD68+, CD45RB+, CD43+ and CD20+ cells were increased in AA as compared with the AL group, most of them significantly. Most of these parameters were also significantly increased in both AA and AL patients as compared with the control group. In both the AA group and the AL group, there existed some significant positive correlations between interstitial tryptase-positive cells and interstitial expression of alpha-SMA, interstitial volume and CD68+ cells. Interestingly, in AA cases, but not in AL cases, we noted a significant relationship between interstitial tryptase-positive cells and CD43+ cells. Our findings demonstrate that mast cells belong to the constitutive cell types in the interstitium in renal amyloidosis, in particular in amyloid type A. In addition, in both the AA group and the AL group, the significant positive correlations between interstitial mast cell count and relative interstitial volume and interstitial expression of alpha-SMA suggest that these cells play a role in the development of interstitial fibrosis.

Quantitative Analysis of the Interstitial Myofibroblasts in Idiopathic Mesangiocapillary Glomerulonephritis Type I

Fourteen renal biopsy specimens from patients with mesangiocapillary glomerulonephritis type I (MCGN-I), for whom both light and electron microscopies as well as immunofluorescence microscopy and full clinical data were available, were examined quantitatively. As a control, 10 biopsy specimens of the kidney from patients with minimal change disease (MCD) were used. Morphometric investigations were performed by a computer image analysis system to investigate whether myofibroblasts have a role in tubulointerstitial fibrosis in MCGN-I and, in particular, to examine the relationship between alpha-SMA expression and interstitial infiltrates. The morphometric study revealed that in MCGN-I patients the mean values for the expression of alpha-SMA, interstitial volume, CD68+, CD45RB+, CD43+ and CD20+ cells were significantly increased, as compared with a control group. In the MCGN-I group, there were significant positive correlations between the interstitial expression of alpha-SMA and the interstitial volume as well as CD68+ and CD45RB+ cells. The correlations between the interstitial expression of alpha-SMA and CD43+ as well as CD20+ cells were positive, but without statistical significance. In conclusion, our study suggests that interstitial a-SMA positive cells play a role in the development of interstitial fibrosis in MCGN-I. The significant correlation between the interstitial expression of alpha-SMA and interstitial CD68+ cells may identify monocytes/macrophages as important mediators in the process of inducing the myofibroblast phenotype in resting fibroblasts.

Tubular NF-κB is overexpressed in proteinuric patients with IgA nephropathy.

Increasing evidence suggests that nuclear factor κB (NF-κB) plays a pivotal role in many glomerulopathies. Therefore, the aim of the present study was to determine the tubular immunoexpression of NF-κB in non-proteinuric (n = 22) and proteinuric patients (n = 16) with IgA nephropathy (IgAN). Another purpose of this study was to examine the possible relationship between NF-κB immunoexpression and proteinuria, interstitial fibrosis as well as interstitial infiltrates. Tubular immunoexpression of NF-κB, interstitial monocytes/macrophages, T lymphocytes, B lymphocytes and interstitial area were determined using a computer image analysis system. The mean values of the tubular immunoexpression of NF-κB, interstitial area and interstitial monocytes/macrophages were in proteinuric IgAN patients significantly increased compared to non-proteinuric IgAN cases, whereas interstitial T and B lymphocytes did not differ between these groups. In proteinuric patients, tubular immunoexpression of NF-κB was highly significantly positively correlated with the degree of proteinuria. Moreover, in both the non-proteinuric and the proteinuric groups with IgAN, tubular immunoexpression of NF-κB was positively correlated with the interstitial area and interstitial monocytes/macrophages. Our findings raise the possibility that proteinuria causes tubular overexpression of NF-κB and, in the process, recruitment of monocytes/macrophages and tubulointerstitial injury in IgAN patients.

Prognostic significance of apoptosis in laryngeal cancer. A quantitative immunomorphological study

The main purpose of the study was to investigate the prognostic significance of apoptosis of cancer cells and to examine the relationship between apoptosis and morphological markers of the host immune response in laryngeal cancer. Apoptotic tumour cells, detected by the TUNEL technique, expression of HLA DR (an antigen belonging to human leukocyte-associated antigens class II) in cancer cells, the number of tumour infiltrating T cells (CD45RO+ cells), B cells (CD20+ cells), macrophages (CD68+ cells) and mast cells as well as the mitotic index were investigated in 28 laryngeal cancers. Sections were studied morphometrically using image analysis. Significant inverse correlations between the number of apoptotic tumour cells and HLA DR-positive tumour cells as well as between the number of apoptotic tumour cells and the number of CD45RO+ cells at the tumour periphery were observed. Analysis of survival showed that patients with high rates of apoptosis had significantly worse prognosis as compared to patients with low apoptotic rates. Differences in HLA DR expression and numbers of CD45RO+ cells were also found between groups of patients with high and low numbers of apoptotic cells. Whether these findings are due to immunosuppressive effects of apoptosis needs further investigation.

A study of apoptosis in human glomerulonephritis as determined by in situ non-radioactive labelling of DNA strand breaks.

It is postulated that programmed cell death via apoptosis may be critical for successful remodelling of glomeruli after inflammation injury. Recently, several methods of analysis of apoptotic cells have been developed, most of them based on the detection of DNA degradation. To determine whether apoptosis is present in renal tissue we compared 36 renal biopsy specimens in patients with various types of glomerulonephritis with 8 samples of the normal kidney using the In situ Cell Death Detection kit, POD, which detects single cell apoptosis in formalin-fixed tissue. In addition, we tried to correlate apoptosis with the type of renal injury. The apoptotic cells were scored using a computerized digital image analyzer. There were greater numbers of glomerular apoptotic cells in IgA nephropathy (IgA-N) compared with the normal kidney, minimal change nephrotic syndrome (MCNS) and mesangial proliferative glomerulonephritis (Mes-Pro-GN). In proliferative types of glomerulonephritis the intensity of apoptosis in glomeruli correlated significantly with the total number of glomerular cells. The relationship between apoptotic cells in glomeruli and degree of proteinuria showed statistical significance only in patients with Mes-Pro-GN. There was no difference in the number of interstitial and tubular apoptotic cells between all studied groups. Our data suggest the possibility of altered regulation in apoptotic mode of cell death in mechanisms of glomerular damage.