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

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

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

We studied the mechanism and inhibition of cell death by exposure to UV alone or combination of UV exposure and intracellular zinc depletion with TPEN in cultured human retinal pigment epithelial cells (RPE). Cell death was quantified by measuring LDH release from injured cells. RPE were exposed to UV at 253.7 nm for 1~20 minutes.The 2~5 minutes UV exposure was duration-dependently cytotoxic, whereas 1minute exposure was minimally so.And exposure to TPEN induced concentration-dependent cell death at 1~4 micrometer range ;0.5 micrometer TPEN was minimally toxic.Then, cultures were exposed to varying exposure durations of UV in the absence or presence of 0.5 micrometer TPEN.At any point, the presence of TPEN markedly increased UV toxicity.In contrast, cell membrane-impermeable zinc chelator showed no toxicity.On the other hand, addition of a protein synthesis inhibitor or caspase inhibitor was markedly protective.In addition, RPE injury with exposure to combination of UV 1 min and TPEN 0.25 micrometer was accompanied by TUNEL and Hoechst staining positivity indicating that the toxicity is mainly apoptosis.Electron microscopic examinations revealed that nuclear fragmentation occurred even in sublethal UV or TPEN exposure, suggesting that the injury process already began at these condition consistently with the death being apoptosis. The present study has shown that combination of known risk factors may act synergistically to induce ARMD etc.The RPE injury induced by low dose UV and minimal intracellular zinc depletion was inhibited with protein synthesis inhibitor or caspase inhibitor, so these results suggested the possibility of prevention or treatment of RPE dysfunction.
Apoptosis ; Cell Death ; Epithelial Cells* ; Hand ; Humans* ; In Situ Nick-End Labeling ; Retinaldehyde* ; Risk Factors ; Zinc*

The apoptotic effect of bacteria-derived beta-glucan was investigated in human colon cancer cells SNU-C4 using terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, reverse transcription-polymerase chain reaction (RT-PCR) expressions of Bcl-2, Bax, and Caspase-3 genes, and assay of caspase-3 enzyme activity. beta-Glucan of 10, 50, and 100 microg/mL decreased cell viability in a dose-dependent manner with typical apoptotic characteristics, such as morphological changes of chromatin condensation and apoptotic body formation from TUNEL assay. In addition, beta-glucan (100 microgram/mL) decreased the expression of Bcl-2 by 0.6 times, whereas the expression of Bax and Caspase-3 were increased by 3.1 and 2.3 times, respectively, compared to untreated control group. Furthermore, the caspase-3 activity in the beta-glucan-treated group was significantly increased compared to those in control group (P < 0.05). Bacterial derived beta-glucan could be used as an effective compound inducing apoptosis in human colon cancer.
Apoptosis ; Caspase 3 ; Cell Survival ; Chromatin ; Colon ; Colonic Neoplasms ; DNA Nucleotidylexotransferase ; Humans ; In Situ Nick-End Labeling

PURPOSE: Replication-competent adenoviruses (Ads) are promising new modalities for the treatment of cancer. Selective replication of a viral agent in tumor may lead to improved efficacy over non-replicating Ads due to viral multiplication, lysis of the infected cancer cell and spread to surrounding cells. In our previous studies it was shown that the E1B 55 kD-deleted Ad (YKL-1) exhibits tumor specific replication and cell lysis, but with reduced cytolytic effects compared to the wild type adenovirus (Int J Cancer 2000;88: 454-463). Thus, improving the potency of oncolytic Ads remains an important goal for cancer gene therapy. To increase the oncolytic ability of YKL-1, an adenovirus death protein (ADP) gene was reintroduced under the control of a CMV or MLP promoter at the E3 region of the YKL-1, generating an YKL-cADP and YKL-mADP, respectively. MATERIALS AND METHODS: The in vitro cytolytic effect of ADP expressing Ads was evaluated by MTT assay, and the induction of apoptosis by ADP expressing Ads was examined by TUNEL analysis. Finally, the antitumor effect of ADP expressing Ads was demonstrated in C33A xenograft tumor model. RESULTS: The YKL-cADP exerted a markedly enhanced cytolytic effect against H460 and SK-Hep1 cancer cell lines. The TUNEL assay indicated that the ADP-mediated cytotoxicity was largely driven by apoptosis. Finally, the YKL-cADP showed a superior antitumor effect than the YKL-1 or YKL-mADP in C33A xenografts. CONCLUSION: These lines of evidence demonstrate that the YKL-cADP induces efficient cell lysis, which is critical for the addition of therapeutic value to replicating Ads in cancer gene therapy.
Adenosine Diphosphate ; Adenoviridae* ; Apoptosis ; Cell Line ; Genes, Neoplasm ; Heterografts ; In Situ Nick-End Labeling