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Triphosphate Nick End Labeling (triphosphate + nick_end_labeling)
Selected AbstractsTopical dorsal skin immersion in seawater induces apoptosis and proliferation in hairless miceTHE JOURNAL OF DERMATOLOGY, Issue 10 2007Min Hong PAN ABSTRACT Recreational and occupational exposure to seawater (SW), have increased but the effect of SW on skin has not been elucidated. The purpose of present study was to assess the effects of SW immersion on the dorsal skin in hairless mice. Adult hairless mice were individually immersed in SW for 3 h, 6 h and 12 h; then, full-thickness dorsal skin of 2 cm diameter was excised for pathological examination (light microscope), apoptosis detection (terminal deoxynucleotidyl transferase-mediated 2,-deoxyuridine 5,-triphosphate nick end labeling [TUNEL]) and proliferation index evaluation (immunohistochemistry). Normal and normal saline (NS)-immersed skin were used as controls. Histological examination revealed that there were randomly distributed cell deaths, presenting cell shrinkage, condensation of nuclear chromatin and eosinophilic cytoplasm in the epidermis, and neutrophil infiltration in the dermis, after SW immersion. Moreover, TUNEL showed low levels of apoptosis in normal (9.07 ± 0.70%) and NS-immersed skin (9.99 ± 1.22%). There was an apparent increase in the 6-h and 12-h SW immersed groups (29.90 ± 6.85%, P < 0.01; 45.46 ± 6.12%, P < 0.01, respectively). Ki-67 antigen was located in the basal layer of the epidermis and hair follicles, the rates of Ki-67-positive cells were 7.90 ± 1.45% and 7.76 ± 1.52% in normal and NS-immersed skin, respectively, and in the 12-h SW immersed group, the rate of Ki-67-positive cells reached 23.85 ± 4.21% (threefold, P < 0.01). In each group, the rate of apoptosis was higher than that of proliferation. We conclude that SW immersion can cause time-dependent apoptosis and proliferation in the epidermis, and the overall effect of SW immersion is injury to the epidermis. [source] Fas-mediated upregulation of vascular endothelial growth factor and monocyte chemoattractant protein-1 expression in cultured dermal fibroblasts: Role in the inflammatory responseTHE JOURNAL OF DERMATOLOGY, Issue 2 2007Masao FUJIWARA ABSTRACT The Fas,Fas ligand interaction is the most important pathway in starting apoptosis. In addition, several recent reports have emerged documenting non-apoptotic roles for Fas. However, a non-apoptotic role of Fas in dermal fibroblasts remains unknown. The present study investigated whether Fas stimulation not only promotes apoptosis but also stimulates elements of the inflammatory response such as angiogenesis and macrophage infiltration. Fas stimulation was performed by treating cultured human dermal fibroblasts with an agonistic anti-Fas monoclonal antibody (mAb). Anti-Fas mAb-treated fibroblasts showed a significantly greater increase of caspase-3 and caspase-8 activity compared with control fibroblasts. Addition of the anti-Fas mAb induced DNA fragmentation, as confirmed by the DNA ladder assay. Terminal deoxynucleotidyl transferase-mediated 2,-deoxyuridine 5,-triphosphate nick end labeling (TUNEL) staining showed that treatment with the anti-Fas mAb induced an increase of apoptotic fibroblasts in a time-dependent manner. At both mRNA and protein levels, anti-Fas mAb-treated fibroblasts showed significantly higher expression of vascular endothelial growth factor (VEGF) and monocyte chemoattractant protein (MCP)-1 compared with control fibroblasts. A pan-caspase inhibitor (Z-VAD-FMK) significantly inhibited VEGF and MCP-1 expression. After transplantation of fibroblasts into mice with severe combined immunodeficiency, the nodules derived from anti-Fas mAb-treated fibroblasts showed more abundant neovascularization, increased macrophage infiltration, and more apoptotic cells in comparison with nodules derived from control fibroblasts. The results of both in vitro and in vivo studies confirmed significantly higher angiogenic activity and macrophage chemotactic activity of anti-Fas mAb-treated fibroblasts compared with control fibroblasts. [source] Cytotoxicity and antiangiogenesis by fibroblast growth factor 2,targeted Ad-TK cancer gene therapy,THE LARYNGOSCOPE, Issue 4 2009Koichiro Saito MD Abstract Objectives: Human head and neck squamous cell carcinoma (HNSCC) in addition to lung, skin, ovarian, and other cancers overexpress fibroblast growth factor (FGF) receptors on both individual tumor cells and endothelial cells within the tumor microenvironment. The purpose of this study was to investigate whether FGF2-targeted gene therapy could redirect adenoviral vectors encoding the herpes simplex virus thymidine kinase gene (Ad-TK) to FGF receptors on tumor and endothelial cells with the intent of improving both the efficiency of transgene expression and the antitumor response. Study Design and Methods: An Ad-TK vector consisting of a conjugate of FGF2 linked to a Fab, fragment against the adenoviral knob region was directly delivered to human HNSCC xenograft tumors in nude mice, which were subsequently dosed with ganciclovir. Tumor specimens were assessed for herpes simplex virus thymidine kinase (HSV- tk) transgene mRNA expression, FGF1/2 receptor expression, terminal deoxynucleotidyl transferase biotin,deoxy uridine triphosphate nick end labeling assay for apoptosis, CD31 immunohistochemistry to estimate tumor microvessel density, and tumor volume change. Results: FGF2-retargeted Ad-TK gene therapy demonstrated significant increases in both HSV- tk mRNA expression and cellular apoptosis levels, and a significant decrease in tumor volume size compared with all other groups. Furthermore, microvessel density was significantly lower in the FGF2-retargeted Ad-TK group, indicating a strong antiangiogenesis effect. Conclusions: These data suggest that FGF2-retargeted Ad-TK produces a combination of expected direct antitumor cytotoxicity and a newly reported antiangiogenesis effect that could prove promising as a novel therapeutic approach in the treatment of FGF receptor,expressing cancers. Laryngoscope, 2009 [source] Programmed cell death in varicocele-bearing testesANDROLOGIA, Issue 1 2009A. Hassan Summary Accelerated apoptosis is a significant factor in the pathophysiology of male infertility disorders associated with abnormal spermatogenesis. This study aimed to investigate apoptosis in varicocele-bearing testes. Sixty four men with varicocele (18 fertile and 46 infertile) were studied compared with eight men with obstructive azoospermic as controls. Apoptosis was assessed in testicular biopsy specimens using terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) method as well as electron microscopy. The results demonstrated that the occurrence of apoptotic changes comprised all types of germ cells but not affecting Sertoli cells. Mean tubular apoptotic indices of fertile or infertile men with varicocele were significantly higher than controls (mean ± SD 4.55 ± 1.03%, 6.29 ± 1.82% versus 2.71 ± 0.45%, P < 0.05). Mean Leydig cells apoptotic indices of infertile men with varicocele were significantly higher than those of fertile men without varicocele as well as controls (1.18 ± 0.38%, 0.68 ± 0.15%, 0.31 ± 0.21%, P < 0.05). Apoptotic indices were nonsignificantly correlated with Johnsen score, testicular volume or varicocele grade. It is concluded that testicular apoptosis is increased in varicocele-associated men either fertile or infertile who may be implicated in associated spermatogenic dysfunction. [source] | ||||||||||