No abstract available.
Animals* ; Brain* ; Hypothermia* ; In Situ Nick-End Labeling* ; Models, Animal*

PURPOSE: Although immunohistochemically detectable metallothionein (MT) overexpression has been described in proliferation epithelial tumor cells, the clinical significance of the expression remains to be elucidated. Therefore, the present article is focused on evaluating the possible significance of MT expression in colonic adenocarcinoma and its relationship with p53 overexpression, Topoisomerase II-alpha as new cell proliferating marker and apoptosis. METHODS: The following formalin-fixed paraffin embedded surgical or biopsied samples were immunohistochemically stained for MT, p53 and topoisomerase II-alpha, and performed in situ TUNEL method for evaluation of apoptotic cell ; normal control mucosa (78 cases), tubular adenomas (20 cases) and adenocarcinomas with various degree of differentiation (78 cases). RESULTS: The MT immunohistochmical reactivity was decreased in colonic adenocarcinoma than that of normal glandular epithelial and tubular adenoma, with the frequency of MT expression in colonic adenocarcinoma depending upon tumor differentiation only. But the frequency of p53 expression was correlated with T-stage, lymph node metastasis and clinical staging, while topoisomerase II-alpha expression and apoptosis in colonic adenocarcinoma were correlated with lymph node metastasis and clinical staging. The immunohistochemical expression of MT and p53 expression in colonic adenocarcinoma was inversely correlated. Also, the inverse correlation between MT expression and expression of toposiomerase II-alpha indices and apoptotic indices were noted. CONCLUSION: These data suggest that MT expression may play a role in proliferative activity and apoptosis in colonic adenocarcinoma. Although MT expression is correlated to tumor differentiation, further studies of a possibility of prognostic factor, such as p53, are required for the determination of significant relationships in other clinicinopathologic indices.
Adenocarcinoma* ; Adenoma ; Apoptosis* ; Colon* ; In Situ Nick-End Labeling ; Lymph Nodes ; Metallothionein* ; Mucous Membrane ; Neoplasm Metastasis ; Paraffin

PURPOSE: The purpose of this study was to determine whether Fas-L expression is associated with increased apoptotic induction of tumor-infiltrating lymphocytes (TIL) in human gastric carcinomas. MATERIALS AND METHODS: The author analysed 38 cases of early gastric carcinoma (EGC) and 61 cases of advanced gastric carcinoma (AGC) who received gastric resection, in whom the number of diffuse type was 38 cases and the number of intestinal type was 61 cases. The author used immunohistochemical staining for Fas, Fas-L and CD45, and TUNEL in situ apoptosis detection kit. TIL were detected by CD45 and apoptosis of TIL were detected by CD45 expression and TUNEL positivity on serial histologic sections. RESULTS: Fas-L was localized to neoplastic cells in 61% (23/38) of EGC group and 66% (40/61) of AGC group. The extent of Fas-L expression was variable, with both Fas-L positive and negative neoplastic region occuring within tumors. TIL adjacent to Fas-L expressing tumor region were decreased in number and TIL adjacent to FasL-negative tumor region were increased in number; apoptotic induction of TIL showed just the opposite pattern (p<0.05). Fas expression was found essentially homogeneously throughout the tumor mass independent of tumor stage. Fas expression showed 64% (39/61) of intestinal type and 68% (26/38) of diffuse type. Labeling indices for tumoral apoptosis in EGC and AGC were 6.72% and 7.13%, respectively and this difference was statistically insignificant. Co-expression of Fas-L and Fas, which occurred over large areas of the tumors, did not result in an enhanced rate of tumor cell apoptosis. In addition, factors such as tumor stage and other prognostic factors were not concerned in Fas and Fas-L expression, number of TIL and apoptotic induction. CONCLUSION: These findings suggest Fas-mediated apoptotic depletion of TIL in response to Fas-L expression by stomach cancers, and provide the evidence to support the Fas counterattack as a mechanism of immune escape in gastric cancer. In addition, gastric carcinoma cells of the intestinal and diffuse type did not differ in their expression of the apoptotic receptor Fas.
Apoptosis* ; Humans ; In Situ Nick-End Labeling ; Lymphocytes, Tumor-Infiltrating ; Stomach Neoplasms ; United Nations

BACKGROUND: The studies on cryopreserved arterial allograft have been focused on cooling methods, pre-treatment, cryoprotectant agents, and preservation temperature. But recently, several studies have reported that thawing methods also play an important role in the occurrence of macroscopic and microscopic cracks. This study was designed to investigate the cell injury after thawing, using a rabbit model to clarify the effect of thawing methods on cryopreserved arteries. MATERIAL AND METHOD: Segments of the rabbit aorta were obtained and divided into 3 groups (n=60) according to whether the specimens were fresh (control, n=20), cryopreserved and rapidly thawed (RT) at 37oC (n=20), or cryopreserved and subjected to controlled, automated slow thawing (ST)(n=20). Cell damage was established using the TUNEL method and the morphological changes were also evaluated. RESULT: In the group that was rapidly thawed, the expression of TUNEL (+) cells increased significantly more than in the slowly thawed group. In addition, the endothelial denudation, microvesicles and edema were significant in the rapidly thawed group compared with those changes in the slowly thawed group. CONCLUSION: Our study suggests that the rapid thawing method may be one of the major causes of cellular damage and delayed rupture in cryopreserved arterial allografts. The expression of TUNEL (+) cells and structural changes were significantly low in the slowly thawed group, which might have contributed to the improvement of graft failure after transplantation.
Allografts ; Aorta* ; Arteries ; Cryopreservation ; Edema ; In Situ Nick-End Labeling ; Rupture ; Transplants

The purpose of this study was to determine the role of apoptosis in the contact lens-worn cornea and the pathophysiologic influence of the contact lens to the rabbit corneal tissue, We had 4 experimental groups; soft contact-worn, RGP contact-worn, mechanically scraped and ormal control groups. The corneas were prepared for routine H & E staining and apoptosis evaluation. Keratocyte and epithelia cell morphology of the cornea were examined in each group using light microscopy. Nuclear DNA fragmentation was detected with the TUNEL assay for 3`-hydroxy DNA ends. The apoptosis assay demonstrated: (a) both the normal cornea and the contact lens-worn cornea exhibited no apoptosis, (b) silight degree of apoptosis was corneal apoptosis detected n deratocytes of the soft contact lensworn cornea, and (c) the anterior stromal keratocytes were found to be frequently undergoing apoptotic change in the scraped cornea. Theses findings suggest that the possible hypoxia induced by soft contact lens-wearing may have a role in apoptosis of anterior stromal keratocytes. To be clinically significant, we need more evaluations and long term studies of apoptosis in contact lens-worn cornea.
Anoxia ; Apoptosis* ; Cornea* ; DNA ; DNA Fragmentation ; In Situ Nick-End Labeling ; Microscopy

The authorsinvestigated the effects of UV-B and amniotic membrane graft about PRK induced inflammatory cell infiltration into corneal stroma, lipid peroxidation and keratocyte apoptosis. Total 20 white rabbits were divided into 5 groups; 1)mechanical epithelial removal, 2)epithelial removal and UV-B irradiation, 3)PRK only, 4) PRK and UV-B irradiation, 5)Amniotic membrane graft after PRK and UV-B irradiation. All corneas were harvested after 24hrs. H & E stain for PMNs infiltration, MDA immunohistochemical stain for lipid peroxidation and TUNEL stain for keratocyte apoptosis were performed. UV-B had little effect on infiltration of inflammatory cell into corneal stroma, lipid peroxidation and keratocyte apoptosis. Amniotic membrane suppressed infiltration of PMNs into corneal stroma, lipid peroxidation and keratocyte apoptosis. Environmental UV-B exposure should not be avoided after PRK. Amniotic membrane graft is beneficial to reduce keratocyte apoptosis and related corneal haze.
Amnion* ; Apoptosis* ; Cornea ; Corneal Stroma ; In Situ Nick-End Labeling ; Inflammation* ; Lipid Peroxidation* ; Membranes ; Rabbits ; Transplants

BACKGROUND: Pilomatricoma is a benign epidermal appendage tumor with differentiation toward hair matrix cells. Histologically, pilomatricoma comprises masses of immature basophilic cells, few transitional cells, and clusters of shadow cells. The mechanism leading to the formation of shadow cells is still unknown. OBJECTIVE: The aim of this study is to examine the expression of p53, bcl-2, Ki-67, and apoptotic rate for the investigation of the cell turnover and cell differentiation within pilomatricoma. METHODS: Immunohistochemical staining(LSAB technique) using monoclonal antibodies including bcl-2, p53, and Ki-67(MIB-1) is performed on skin biopsy specimens of pilomatricoma, and TUNEL staining for detecting apoptotic cells is also performed. RESULTS: The expression of Ki-67 and bcl-2 is noted in basal basophilic cells more than overlying basophilic cells. The p53 protein is observed to be alike on basal and overlying basophilic cells. But apoptotic cells are only expressed in transitional cells. CONCLUSION: The result of this study suggests that high proliferative area, such as basal basophilic cells manifested by over-expression of Ki-67 and bcl-2, is regulated by the p53 protein inducing apoptosis. Thereafter, basophilic cells may progress to shadow cells through apoptotic transitional cells by the action of p53 protein.
Antibodies, Monoclonal ; Apoptosis* ; Basophils ; Biopsy ; Cell Differentiation ; Hair ; In Situ Nick-End Labeling ; Pilomatrixoma* ; Skin

BACKGROUND: The cause of acrodermatitis enteropathica(AE) is closely related to zinc deficiency. Zinc is a potent inhibitor of endonuclease. Acute rises in the apoptosis in lymphoid and myeloid cell lines during zinc deficiency has recently been reported. The method of terminal transferase mediated dUTP biotin nick end labeling(TUNEL) is used in situ labelling of apoptotic nuclei in routine tissue sections. OBJECTIVE: The purpose of this study is to clarify our hypothesis that apoptosis resulted from zinc deficiency might cause keratinocytes damages in AE. METHOD: We stained 6 AE biopsy specimen with TUNEL technique. RESULTS: In acroderrratitis enteropathica, apoptotic keratinocytes were shown in the entire epidermis as compared to normal, controlled skin, in which it was found only at the uppermost layer of this stratified epithelium. CONCLUSION: This result suggests that apoptosis resulting from zinc deficiency might play a role in keratinocyte death in AE.
Acrodermatitis* ; Apoptosis ; Biopsy ; Biotin ; Epidermis ; Epithelium ; In Situ Nick-End Labeling ; Keratinocytes* ; Myeloid Cells ; Skin ; Transferases ; Zinc

PURPOSE: The study was aimed to detect the differences in the cell viability and the apoptosis induction after irradiation on normal and tumorigenic cells. MATERIALS AND METHODS: The study, that was generated for two human normal cells(RHEK, HGF-1) and two human tumor cells(KB, HT-1080), was tested using MTT assay at 1 day and 3 day after irradiation and TUNEL assay under confocal laser scanning microscope at 1 day after irradiation. Single irradiation of 0.5, 1, 2, 4, and 8 Gy were applied to the cells. The two fractions of 1, 2, 4, and 8 Gy were separated with a 4 hour time interval. The irradiation was done with 5.38 Gy/min dose rate using Cs-137 irradiator at room temperature. RESULTS AND CONCLUSIONS: 1. In 3-day group, the cell viability of HGF-1 cell was significantly decreased at 2, 4 and 8 Gy irradiation, the cell viability of KB cell was significantly decreased at 8 Gy irradiation and the cell viability of HT-1080 cell was significantly decreased at 4 and 8 Gy irradiation. 2. There was significant difference between RHEK and KB cell line in the cell viability of 3-day group at 8 Gy irradiation. There was significant difference between RHEK and HGF-1 cell line in the cell viability of 3-day group at 4 and 8 Gy irradiation. 3. There was a significantly decreased cell viability in 3-day group than those in 1-day group at 2, 4 and 8 Gy on HGF-1 cell, at 4 and 8 Gy on HT-1080 cell, at 8 Gy on KB cell. 4. We could detect DNA fragmented cells only on KB cell. Number of apoptotic cells of KB cell was significantly increased at 4 and 8 Gy irradiation. However, there was no correlation between cell viability and apoptosis. 5. On all 4 cell lines, there were no differences between single and split irradiation method in cell viability and apoptosis.
Apoptosis* ; Cell Line ; Cell Survival* ; DNA ; Humans ; In Situ Nick-End Labeling ; KB Cells ; Radiation Dosage*

PURPOSE: This study was performed to investigate apoptosis by radiation in the developing fetal rat brain. MATERIALS AND METHODS: Fetal brains were irradiated in utero between the 17th and 19th days of fetal life (E17-19) by linear accelerator. A dose of irradiation ranging from 1 Gy to 4 Gy was used to evaluate dose dependency. To test time dependency the rats were irradiated with 2 Gy and then the fetal brain specimens were removed at variable time course; 1, 3, 6, 12 and 24 hours after the onset of irradiation. Immunohistochemical staining using in situ TdT-mediated dUTP nick end labelling (TUNEL) technique was used for apoptotic cells. The cerebral cortex, including three zones of cortical zone (CZ), intermediate zone (IZ), and ventricular zone (VZ), was examined. RESULTS: TUNEL positive cells revealed typical features of apoptotic cells under light microscope in the fetal rat cerebral cortex. Apoptotic cells were not found in the cerebral cortex of non-irradiated fetal rats, but did appear in the entire cerebral cortex after 1 Gy irradiation, and were more extensive at the ventricular and intermediate zones than at the cortical zone. The extent of apoptosis was increased with increasing doses of radiation. Apoptosis reached the peak at 6 hours after the onset of 2 Gy irradiation and persisted until 24 hours. CONCLUSION: Typical morphologic features of apoptosis by irradiation were observed in the developing fetal rat cerebral cortex. It was more extensive at the ventricular and intermediate zones than at the cortical zone, which suggested that stem cells or early differentiating cells are more radiosensitive than differentiated cells of the cortical zone.
Animals ; Apoptosis* ; Brain ; Cerebral Cortex* ; In Situ Nick-End Labeling ; Particle Accelerators ; Rats* ; Stem Cells