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

BACKGROUND: Recently, the pulse-in-pulse mode of intense pulsed light (IPL) has been used increasingly for the treatment of melasma. OBJECTIVE: To observe the morphologic changes in the melanophore in adult zebrafish after irradiation with conventional and pulse-in-pulse IPL and Q-switched Nd:YAG (QSNY) laser. METHODS: Adult zebrafish were irradiated with conventional and pulse-in-pulse mode of IPL. The conditions for conventional IPL were 3 mJ/cm², 560 nm filter, and pulse widths of 7, 20, and 35 msec. The pulse-in-pulse conditions were 3 mJ/cm² and on-time 1/off-time 2. The QSNY laser was used with the settings of 1,064 nm, 0.4 J/cm², a 7 mm spot size, and one shot. Specimens were observed using a light microscope, a transmission electron microscope (TEM), a scanning electron microscope (SEM) and a confocal microscope. RESULTS: After conventional IPL irradiation with a 7 msec pulse width, melanophore breakage was observed using light microscopy. Under TEM, irradiation with conventional IPL for 7 msec and pulse-in-pulse IPL induced melanophore thermolysis with vacuolization. However, changes in the melanophore were not observed with 35 msec IPL. Under SEM, unlike the control and QSNY groups, IPL-irradiated zebrafish showed finger-like fusion in the protein structure of scales. Specimens examined by a confocal microscope after conventional IPL irradiation showed a larger green-stained area on TUNEL staining than that after pulse-in-pulse mode IPL irradiation. CONCLUSION: Zebrafish irradiated with long pulse-IPL showed no morphologic changes using light microscopy, while morphological changes in melanophores were evident with use of TEM. Pulse-in-pulse mode IPL caused less damage than conventional IPL.
Adult ; Humans ; In Situ Nick-End Labeling ; Melanophores* ; Melanosis ; Microscopy ; Weights and Measures ; Zebrafish*

PURPOSE: This study was aimed to evaluate changes in the stromal keratocyte after ablation of 50 micrometer and 100 micrometer with use of photorefractive keratectomy(PRK). METHODS: At 4 hours, 24 hours, 72 hours, 7 days and 1 month after PRK, each group of rabbits including normal control group was treated with terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling(TUNEL) staining using ApopTag(R) kit in vivo, then apoptotic keratocytes were evaluated with light microscope. RESULTS: There was no response with TUNEL staining of the epithelial cells, stromal keratocyte, and endothelium in normal cornea. In the ablation group, however, regardless of the depth of photorefractive ablation, the TUNEL signal was maximal after 4 hours, and it decreased with time. The signal was more intense in 100 micrometer ablation group than 50 micrometer ablation group, although the signal was not observed at the endothelial cells in both groups. The number of apoptotic stromal keratocytes at each time point of 4 hr, 24 hr, 72 hr, and 1 week was 57+/-8.9, 49+/-7.5, 36+/-5.1, and 12+/-1.3 cells/field in 100 micrometer ablation group, and 31+/-4.4, 28+/-4.6, 21+/-3.9, and 5+/-1.1 cells/field in 50 micrometer ablation group, and the difference between the two groups was statistically significant(P<0.05). CONCLUSION: The more the amount of ablation with photorefractive keratectomy, the stronger the apoptotic response. The postoperative apoptotic response was observed especially within 1 week. These findings suggest that early suppression of postoperative apoptosis within 1 week will influence on the prognosis of visual quality after photorefractive keratectomy, and more studies will be needed in the future.
Apoptosis* ; Cornea ; Endothelial Cells ; Endothelium ; Epithelial Cells ; In Situ Nick-End Labeling ; Photorefractive Keratectomy* ; Prognosis ; Rabbits

We have studied morphologic characteristics and apoptosis on the fetal brain of the trisomy 16 mouse, a model for human trisomy 21 syndrome. This study was based on serial sections of the whole brain from a sample of sixteen trisomy 16 mice and forty-six age-matched control littermates from embryonic day (ED) 12 to ED 18. Trisomy 16 brains showed a reduction of telencephalic size and abnormal cortical development. At ED 13 trisomy 16 and control brains appeared similar. By ED 14 difference in the cortical thickness and telencephalic growth became evident, and by ED 16 a marked size difference had developed between the trisomy 16 and control brains. By ED 18, however, the thickness of the trisomy 16 cortex had increased considerably and was not significantly different with respect to the thickness and cross-sectional areas of the pallium and its constituent cortical layers. The cell density of the trisomy 16 cortex had persistently decreased before ED 17, when the cell density of control and trisomy 16 corteces was similar within each layer. At ED 18 cell density of trisomy 16 cortex in each layer increased. There was inverse relationship between a number of TUNEL positive apoptotic cells and cell density in the trisomy 16 brains. Our results suggest that developmental abnormalities of the trisomy 16 brain indicated developmental delay of the telencephalon growth, which may be caused by apoptosis rather than by a proliferation defect.
Animals ; Apoptosis* ; Brain* ; Cell Count ; Down Syndrome ; Humans ; In Situ Nick-End Labeling ; Mice* ; Telencephalon ; Trisomy*

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

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 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

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

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