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

PURPOSE: To evaluate protective mechanism of melatonin against radiation damage and its relationship with apoptosis in mouse jejunum. MATERIALS AND METHODS:' 168 mice were divided into 28 groups according to radiation dose and melatonin treatment. To analysis crypt survival, microcolony survival assay was done according to Withers an (l Elkind's method. To analysis apoptosis, TUNEL assay was done according to Labet-Moleur's method. RESULTS: Radiation protection effect of melatonin was demonstrated by crypt survival assay and its effect was stronger in high radiation dose area. Apoptosis index with 8 Gy irradiation was 18.4% in control group and 16.5% in melatonin treated group. After 18 Gy, apoptosis index was 17.2% in control group and 15.4% in melatonin treated group. Apoptosis index did not show statistically significant difference between melatonin treated group and control group. CONCLUSION: Melatonin shows clear protective effect in mouse jejunum against radiation damage but it.', protective effect seems not to be related with apoptosis protection effect.
Animals ; Apoptosis* ; Cell Survival* ; In Situ Nick-End Labeling ; Jejunum ; Melatonin* ; Mice* ; Radiation Protection

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*

This study evaluated the response of the anterior stromal keratocytes in Rabbits following deepithelialization and 3 diopter(37micrometer)- and 12 diopter(99micrometer) PRK. The corneal sections obtained from the operated area on postoperative 3, 7 and 14 days were stained with hematoxylin and eosin. Keratocyte apoptosis were monitored using the terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling(TUBEL) staining with ApopTag kit for the corneal section obtained on postoperative 3 day. The corneal haze on postoperative 14 day were graded using a slit lamp biomicroscopy. The number of anterior stromal keratocytes had decreased significantly and positive TUNEL staining was noted in the anterior stroma after PRK and deepithelialization campared with that of controls. The decreased keratocyte numbers were recovered on postoperative 7 day after deepithelialization and on postoperative 14 day after PRK. The newly appeared deratocytes were pyknotic, variable-shaped and crosswisely oriented in appearance, and especially increased following 12 diopter PRK. Both the keratocyte loss and corneal haze grading was increased related to the increased ablation depth after PRK. In conclusion, the loss of anterior stromal keratocytes after PRK is mediated by apoptosis and followed by reactive cellular proliferation might be a important role in the corneal haze.
Apoptosis ; Cell Proliferation ; Cornea* ; Eosine Yellowish-(YS) ; Hematoxylin ; In Situ Nick-End Labeling ; Photorefractive Keratectomy* ; Rabbits

p27kip1, a cyclin dependent kinase inhibitor, has been recognized as a negative regulator of cell cycle. To investigate the role of p27kip1 on progression of cancer and apoptotic pathway, we analyzed p27kip1 expression using immunohistochemical stain in 40 cases of prostatic adenocarcinoma and apoptotic index by TUNEL method in 30 cases of prostatic adenocarinoma. Both were correlated with Gleason grade and Gleason score. Loss of p27kip1 expression was more frequent in prostatic adenocarcinomas of higher score (Gleason score 7 to 10) (60.7%) than in those of lower score (Gleason score 4 to 6) (33.3%) (p<0.05). The value of mean apoptotic index of carcinoma was 1.13+/-0.26, 1.80+/-0.91, 2.06+/-0.79, and 2.12+/-0.82 in grade 2, 3, 4, and 5, respectively, and was positively correlated with grade of carcinoma (p<0.05). Mean apoptotic index of higher Gleason score (score 7 to 10; 2.05+/-0.63) was also significantly increased than in lower Gleason score (score 4 to 6; 1.34+/-0.39) (p<0.05). Mean apoptotic index in cases with and without p27kip1 expression was 1.92+/-0.86 and 1.89+/-0.81, respectively (p>0.05). These results suggest that loss of p27kip1 expression and increased apoptotic index may be the morphologic markers to predict the behavior of prostatic adenocaricnoma. The role of p27kip1 on apoptotic pathway seems to be meager in this study and needs further study.
Adenocarcinoma* ; Cell Cycle ; Cyclins ; In Situ Nick-End Labeling ; Neoplasm Grading ; Phosphotransferases

PURPOSE: To investigate the efficacy of an amniotic membrane contact lens on corneal epithelial wound healing. METHODS: We made a model with a corneal epithelial wound by applying 6 mm round filter paper soaked with 1 N NaOHonto the central cornea in 24 eyes of 12 rabbits. The rabbits were divided into three groups: AMCL (amniotic membrane contact lens), T-AMT (temporary amniotic membrane transplantation) and the control group. We evaluated corneal wound healing every postoperative day using a digital photo slitlamp and fluorescein dye. The corneas were harvested for histopathologic studies after seven days and analyzed with hematoxylin-eosin (H & E) stain and TUNEL staining. RESULTS: The average wound healing time was similar between the amniotic membrane contact lens and the temporary amniotic membrane transplantation group. The number of the infiltrated PMNs (polymorphonuclear cells) was 8.8+/-2.58, 8.6+/-2.19 and 48.6+/-7.12 in the AMCL, T-AMT and control groups, respectively. Apoptotic keratocytes were 3.8+/-1.1, 3.6+/-1.09 and 23.2+/-5.06 in the AMCL, T-AMT and control groups, respectively. In the AMCL and T-AMT groups, the number of infiltrated PMNs and apoptotic keratocytes were significantly less than those the control group (p<0.05). There were not significant differences in the number of PMNs and apoptotic cells in the AMCL and the T-AMT groups. CONCLUSIONS: Amniotic membrane contact lenses have the benefits of being an easily applied method and having a wound healing ability comparable to that possible with conventional suture methods.
Amnion ; Contact Lenses ; Cornea ; Eye ; Fluorescein ; In Situ Nick-End Labeling ; Membranes ; Rabbits ; Sutures ; Transplants ; Wound Healing

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

PURPOSE: Gene-attenuated replication-competent adenoviruses are emerging as a promising new modality for the treatment of cancer. In an effort to continually improve upon cancer gene therapy, we have modified gene- attenuated replication-competent adenoviruses so as to cause them to replicate efficiently and lyse the infected cancer cells more effectively. MATERIALS AND METHODS: We modified the E1 region of the adenovirus (Ad) systematically, generating Ad-deltaE1B19, Ad-deltaE1B55, Ad-deltaE1B19/55, and Ad-WT. The cytopathic effects (CPE) and viral replication of these four gene modified adenoviruses were compared, and the morphology and DNA fragmentation of the infected cells was evaluated. RESULTS: Among the constructed adenoviruses, E1B 19kD-inactivated adenovirus (Ad-deltaE1B19) was the most potent, inducing the largest-sized plaques and markedCPE. Moreover, cells infected with Ad-deltaE1B19 showed complete cell lysis with disintegrated cellular structure whereas cells infected with Ad-WT maintained intact cellular and nuclear membrane with properly structured organelles. TUNEL assay was also used to monitor DNA integrity, and a more profound induction of apoptosis was observed in the Ad-deltaE1B19 infected cells in comparison to wild type adenovirus infected cells. CONCLUSION: We demonstrate that the inactivation of the E1B19kD gene in a replicating adenovirus leads to increased CPE, rapid viral release, improved cell-to-cell viral spread and increased induction of apoptosis.
Adenoviridae* ; Apoptosis ; Cellular Structures ; DNA ; DNA Fragmentation ; Genes, Neoplasm* ; In Situ Nick-End Labeling ; Nuclear Envelope ; Organelles

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

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*