Differential glomerular immunoexpression of matrix metalloproteinases MMP-2 and MMP-9 in idiopathic IgA nephropathy and Schoenlein-Henoch nephritis.

Both idiopathic IgA nephropathy (IgAN) and Schoenlein-Henoch nephritis (SHN) are characterized by cell proliferation and abnormal extracellular matrix (ECM) remodeling by mesangial cells leading to fibrosis, sclerosis and end-stage renal disease. Matrix metalloproteinases MMP-2 and MMP-9 are reported as the most important proteolytic enzymes involved in remodeling of ECM. Therefore, the aim of the present study was to determine glomerular immunoexpression of MMP-2 and MMP-9 in IgAN and SHN. Another purpose of this study was to examine the relationship between expression of MMPs and mesangial cells, a-smooth muscle actin (alpha-SMA) staining, and monocytes/macrophages. Fifteen patients with idiopathic IgAN and 12 with SHN were examined by percutaneous renal biopsy. Glomerular staining intensity of MMP-2 and MMP-9 was recorded semiquantitatively, whereas mesangial cells, glomerular alpha-SMA staining and glomerular CD 68+ cells were assessed quantitatively using computer image analysis system. Our study revealed that the mean values of glomerular immunoexpression of MMP-2, mesangial cells, alpha-SMA staining and glomerular CD 68+ cells were in SHN patients significantly increased as compared to IgAN cases whereas glomerular staining for MMP-9 did not differ in these groups. Moreover, a glomerular staining of MMP-2 was significantly positively correlated with mesangial cells as well as glomerular alpha-SMA staining in both SHN and IgAN. A positive significant correlation between glomerular MMP-2 staining and glomerular CD68+ cells was noted only in SHN group. The correlations of glomerular MMP-9 and these parameters were weak and not significant. In conclusion, our results confirm increased glomerular staining of MMP-2 but not MMP-9 in SHN patients. A suggestion that augmented mesangial cells proliferation in these cases depends on MMP-2, alpha-SMA and monocytes/macrophages needs further investigations including double staining study

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

Human papillomavirus in sinonasal inverted papillomas and squamous cell carcinoma of the larynx. In situ hybridization with human papillomaviruses DNA probes

Background and Purpose: a substantial number of studies have reported evidence on the involvement of human papillomaviruses (HPV) in oral and sinonasal papillomas and carcinomas. Oncogenic HPV subtypes 16 and 18 seem to play an important role in carcinogenesis, whereas low risk HPV subtypes 6 and 11 are usually characteristic of benign lesions. The aim of the present study was to evaluate a role for HPV in the development of sinonasal inverted papillomas and squamous cell carcinomas of the larynx. Materials and Methods: A ten sinonasal inverted papillomas and eight squamous cell carcinomas of the larynx were retrieved from archival material. All examined cases showed histological features of HPV infections (koilocytosis, binucleated squamous cells and abnormal mitoses). All cases have been analyzed using in situ hybridization. A wide spectrum probe for HPV was used initially. Further subtyping was carried out using specific probes for HPV low risk and HPV high risk. Results: Of the ten studied cases of inverted papillomas, one was positive for both: wide spectrum probe and low risk probe, and one was positive for HPV high risk probe only. One of the eight cases of carcinomas was positive for both HPVDNAprobes: wide spectrum probe and high risk probe. Additional positive reactions for HPV DNA using high risk probe were noted in three examined cases of squamous cell carcinomas of the larynx. Conclusions: Our study confirms the role of high risk HPV subtypes in carcinomas. In contrast to other studies, one case only suggests the possible involvement of HPV 6 and HPV 11 in etiology of sinonasal inverted papillomas. The data concerning of the role of HPV in transformation inverted papillomas to squamous cell carcinomas still need further confirmation

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. (Folia Histochemica et Cytobiologica 2011; Vol. 49, No. 1, pp. 34–40

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.

Immunohistochemical study on neuropilin 1 (NRP1) immunoexpression in oral squamous cell carcinoma

  Introduction. Neuropilins (NRPs) are multifunctional glycoproteins that play an important role in angiogenesis and cancer progression. The aim of the study was to examine the immunoexpression of neuropilin 1 (NRP1), the number of NRP1+ infiltrating cells and CD163+ macrophages, and density of microvessels (MVD) in oral squamous cell carcinoma (OSCC). Material and methods. The study was performed on 45 OSCC patients with metastases (OSCCM+), 51 patients without metastases (OSCCM-) and 17 control cases. The microvessels were identified by the presence of CD31 and the expression of the studied proteins was assessed by immunohistochemistry. Results. The immunoexpression of NRP1, the mean numbers of NRP1+, CD163+ infiltrating cells, and MVD were significantly increased in OSCCM+ patients in comparison to OSCCM-, and control groups. Moreover, in OSCCM- patients all these parameters were also significantly increased in comparison to controls. In OSCCM+ and OSCCM- groups, there were positive correlations between the immunoexpression of NRP1 and MVD (r = 0.41, p < 0.006; r = 0.51, p < 0.001, respectively), and between the number of NRP1+ infiltrating cells and CD163+ macrophages (r = 0.56, p < 0.001, r = 0.49, p < 0.001, respectively). Conclusions. The present study revealed overexpression of NRP1 in OSCC, especially in OSCC patients with metasta­sis, suggesting that NRP1 could potentially contribute to metastasis of oral cancer. The correlation between the number of NRP1+ infiltrating cells and CD163+ macrophages suggests that NRP1+ infiltrating macrophages are present in tumor microenvironment and may play a role in the progressions of oral cancer

Aberrant Tubulointerstitial Immunoexpression of Matrix Metalloproteinases MMP-2, MMP-9 and Tissue Inhibitor of Matrix Proteinase-2 (TIMP-2) in Acute Cellular Rejection of Human Renal Allograft

Acute cellular rejection (ACR) may initiate chronic allograft dysfunction with alterations in the extracellular matrix compartment (ECM). Turnover of ECM proteins is regulated by matrix metalloproteinases (MMPs). The aim of the present study was to determine the immunoexpression of MMP-2, MMP-9 and TIMP-2 in ACR, and to examine the relationship between expression of MMPs and monocytes/macrophages, transforming growth factor β-1 (TGFβ-1), and α-smooth muscle actin (α-SMA). Immunoperoxidase study with antibodies against MMP-2, MMP-9, TIMP-2, CD68, TGFβ-1 and α-SMA was carried out on 24 renal allograft biopsy specimens from patients with ACR and 11 allograft biopsy specimens from patients with no signs of rejection. Our study revealed increased immunoexpression of MMP-2, MMP-9 and TIM-2 in ACR as compared with controls, and signifi cant positive correlations between immunostaining of MMP-2 and TGF-β-1, as well as between MMP-2 and α-SMA. Increased immunoexpression of MMP-9 was positively correlated with α-SMA, and the number of interstitial CD68+cells. In conclusion our study supports a role of gelatinases in tissue damage in human renal acute cellular allograft rejection and provides some interesting insights into early renal remodeling which may lead to chronic allograft dysfunction

One week’s holiday sun exposure induces expression of photoaging biomarkers

Introduction. Skin aging is accompanied by the upregulation of the expression of various matrix metalloproteinases (MMPs). It was shown that exposure to ultraviolet radiation (UVR) may induce skin expression of MMPs and dysregulation of the transforming growth factor beta (TGF-β)/Smad pathway. The aim of our study was to compare the effects of short holiday UVR exposure and lifetime UVR exposure, on the expression of MMP-8, TGF-β1, and Smad2 in human skin biopsies. Material and methods. Skin biopsies were taken from the outer upper arm of 15 elderly people with significant photoaging (mean age 64.1 years) (Group 1) and from 15 healthy young adult volunteers (mean age 24.1 y) who participated in a six-day sun holiday. Biopsies were taken twice: 24 hours before leaving for holiday (Group 2a) and 24 hours after returning (Group 2b). The expression of TGF-β1, Smad2, and MMP-8 was examined by immunochemistry and measured semiquantitatively by two independent pathologists. Results. The mean expression of TGF-β1 in dermal fibroblasts and keratinocytes in Group 1 and Group 2b was significantly lower than in Group 2a (0.54% ± 0.44% and 0.48% ± 0.51% vs. 1.48% ± 0.72%, respectively). The percentage of Smad2 (+) cells in Group 1 and Group 2b was lower than in Group 2a (2.13% ± 1.39% and 1.81% ± 1.16% vs. 4.13% ± 1.58%, respectively). The MMP-8 expression in Group 2b was 1.36% ± 0.68% and was significantly higher than in Group 1 (0.34% ± 0.42%) and Group 2a in which the protein was not detected (p < 0.001). Conclusions. We conclude that the decrease in the expression of TGF-β1 and Smad2 is a persistent biomarker of skin photoaging, while the increased expression of MMP-8 in keratinocytes can be regarded as a marker of acute sun exposure

Different distribution of c-kit positive interstitial cells of Cajal-like in children’s urinary bladders

We describe the presence of c-kit positive interstitial cells of Cajal-like (ICCs-like) in the walls of the urinary bladders of children. An immunohistochemical study of specimens, obtained at autopsy from either the trigonum (Group A) or the corpus (Group B), was performed using antibodies against c-kit (CD 117). Histological morphometry of the immunoexpression of c-kit positive ICCs-like was performed by means of image analysis system. The c-kit positive ICCs-like were identified by their morphology and counted in the vesical muscle layer in ten adjacent high power fields, each of 0.0479 mm2. The areas of the epithelial and subepithelial layers containing c-kit positive mast cells (rounded body with no dendritic processes) were neglected. The results were expressed as the number of ICCs-like cells per mm2. Differences between groups were tested using unpaired Student&#8217;s t-test preceded by evaluation of normality and Levene&#8217;s test. Results were considered statistically significant if p < 0.05. In Group A, the mean number of ICCs-like cells was statistically significantly higher (41.5 cells/mm2) than in Group B (30.4 cells/mm2), p < 0.05. ICCs-like cells were found within the smooth muscle layer of the urinary bladder. There was a different distribution of these cells in particular parts of the bladder, which was probably due to the different roles of the trigonum and the corpus in the bladders of children. (Folia Histochemica et Cytobiologica 2011; Vol. 49, No. 3, pp. 431&#8211;435

Usefulness of serum VEGF concentration measurement to estimate aortic aneurysm risk of rupture

Wstęp. Celem niniejszej pracy jest ocena przydatności oznaczania wartości stężeń VEGF w surowicy u chorych z tętniakami aorty brzusznej w celu oceny ryzyka pęknięcia. Analizowano stężenia VEGF w surowicy pacjentów z tętniakami aorty w zależności od objawów klinicznych tętniaka oraz zbadano korelację wartości stężeń VEGF w surowicy z ekspresją tego czynnika w ścianie aorty.Materiał i metody. Materiał badawczy stanowiły próbki surowicy pobrane od chorych przed zabiegiem oraz fragmenty ściany aorty uzyskane od tych samych pacjentów podczas operacji rekonstrukcyjnej aorty. Wyodrębniono następujące grupy: I - tętniaki operowane planowo (n = 49), II - tętniaki objawowe i pęknięte (n = 19), III - niedrożności aortalno-biodrowe (n = 17), IV - grupa kontrolna, którą stanowiły fragmenty ściany aorty pobranej od dawców narządów (n = 9) oraz surowica pobrana u chorych operowanych planowo z powodu przepuklin i żylaków kończyn dolnych (n = 34). Stężenie VEGF w surowicy oznaczano metodą ELISA. Ekspresję VEGF w tkance oceniono morfometrycznie, zliczając komórki VEGF-dodatnie po wykonaniu odczynów immunohistochemicznych. Wyniki. Ekspresja VEGF w ścianie aorty była największa w grupie II. Stwierdzono istotną statystycznie różnicę tej wartości w porównaniu z grupą I (p < 0,003). Wykazano statystycznie znamienne różnice ekspresji VEGF między pozostałymi grupami (p < 0,001). Największą średnią wartość stężenia VEGF w surowicy odnotowano w grupie II, ale różnice między wartościami w poszczególnych grupach nie były istotne statystycznie, mimo że korelacja między wartościami VEGF w surowicy a ekspresją w tkance okazała się silna (p < 0,001). Wniosek. Wyniki badań wskazują na istotną rolę VEGF (i angiogenezy) w rozwoju tętniaka aorty, jednak brak specyficzności uniemożliwia zastosowanie pomiaru jego stężeń w ocenie dynamiki powiększania się tętniaka i ryzyka jego pęknięcia.Background. Risk of abdominal aortic aneurysm (AAA) rupture is difficult to estimate. Angiogenesis in aneurysm walls is an important morphologic finding. One of the most important factors stimulating angiogenesis is vascular endothelial growth factor (VEGF). The aim of our study was to evaluate if the two values: VEGF expression in aortic wall tissues, and VEGF serum concentration correlate with clinical manifestations of aneurysms, and if these two values correlate with each other. Material and methods. Aorta tissue samples were taken in the operating room from patients undergoing aorta reconstruction for aneurysms: electively (group I, n = 49), emergency (group II, n = 19) or because of aortoiliac occlusion (AIO) (n = 17). Control tissue was taken from healthy organ donors (n = 9). Blood samples were obtained from these patients before surgery. Control serum samples were taken from patients undergoing surgery because of hernias and varices. Expression of VEGF in tissue was measured with use of morphometric analysis in slides after immunohistochemistry with anti-VEGF antibodies. Vascular endothelial growth factor serum concentration was measured with the use of ELISA. Results. The highest level of serum VEGF was observed in the symptomatic AAA group (mean value: 404.3 pg/ml; sv = 331.7). Electively operated AAA showed lower serum VEGF concentration (mean value = 285.3; sv = 300.9), AIO and control: 366.4 and 277.3 respectively. These differences were not significant. Strong correlation was observed between VEGF serum level and VEGF tissue expression. Significant differences were shown in VEGF positive cell numbers between all examined groups (mean cell number in AAA symptomatic = 140.9, elective AAA = 108.5, AIO = 51.4, control = 21.0). Conclusions. There is strong correlation in VEGF tissue expression with clinical manifestation of AAA. Vascular endothelial growth factor serum concentration is not a good clinical marker to estimate the risk of rupture