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

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

OBJECTIVE: To investigate the change of placental apoptosis and the expression of their mediator in preeclampsia women. METHODS: Placental tissues from 10 cases of preeclampsia and 15 cases of normal pregnancy were analyzed using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling(TUNEL) staining. Expressions of bcl-2, bax, caspase-3 was also assessed using immunohistochemistry. RESULTS: In TUNEL staining, the number of apoptotic nuclei were significantly increased in the trophoblast of preeclampsia than normal pregnancy. Bcl-2 was mainly expressed in syncytiotrophoblast and bax was mainly expressed in cytotrophoblast. Bcl-2 expression was decreased and bax expression was increased in the preeclampsia than normal, but the difference was not significant. Caspase-3 was mainly expressed in the cytotrophoblast and expression was significantly increased in the preeclampsia than normal pregnancy(p<0.05). CONCLUSION: Placental apoptosis, especially accompanied with increased expression of caspase-3 in cytotrophoblast, might be related with in the pathogenesis of preeclampsia.
Apoptosis* ; Caspase 3 ; Deoxyuridine ; Female ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Placenta ; Pre-Eclampsia* ; Pregnancy ; Trophoblasts

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

OBJECTIVE: This study is designed to investigate how apoptosis is presented and how the genes of p53 and bcl-2 are expressed depending on graded injury in experimental spinal cord injury. METHODS: Experimental spinal cord injury was made on rats with weight drop method. Two different amounts of impact were applied on rat spinal cord. Rats were categorized into three groups (control; five rats, mild injury; five rats, severe injury; five rats). Fourty eight hours following cord injury, cord specimen was harvested from injury epicenter. TUNEL staining was done for apoptotic detection and immunohistochemical staining for p53 and bcl-2 expression. Positively stained cells were counted and mean values were compared among three groups. RESULTS: TUNEL positive cells increased depending on injury severity(p=0.027). The p53 positive cells increased in both injury groups compared to control group(p=0.001). Bcl-2 positive cells decreased as injury amount increased(p=0.002). The p53 expression increased in proportion to TUNEL staining in correlation curve in white matter(correlation coefficient, 0.387). The bcl-2 expression was inversely proportional to TUNEL staining and steeper decrease was found in gray matter than in white matter (correlation coefficient, -0.875). CONCLUSION: Apoptosis increases as the injury grading elevated within 20gm-cm of impact. The p53 seems to promote apoptosis in white matter, but do not show proportional relationship with injury amount. Bcl-2 appeared to be protective to cell death due to apoptosis.
Animals ; Apoptosis* ; Cell Death ; Contusions* ; In Situ Nick-End Labeling ; Rats* ; Spinal Cord Injuries ; Spinal Cord*

OBJECTIVE: To verify the effect of Matrigel, a ECM complex from Engelbreth-Holm-Swarm(EHS) mouse sarcoma on the preimplantation development and apoptosis of mouse fertilized eggs. METHOD: Late Pronucleus stage eggs were cultured through the blastocyst stage in the presence of Matrigel (0.5%, v/v). Characteristics of apoptosis and cell number assessed by Hoecst staining and TUNEL labeling at the blastocyst stage, respectively. RESULTS: Morphological development, number of cells per embryo was significantly increased but rate and number of TUNEL positive nuclei of the embryo were decreased in the presence of Matrigel. CONCLUSION: This result suggested that at low concentration of Matrigel improves both viability and morphological development in the preimplantation mouse embryos.
Animals ; Apoptosis* ; Blastocyst ; Cell Count ; Eggs ; Embryonic Structures* ; In Situ Nick-End Labeling ; Mice* ; Ovum ; Sarcoma ; Zygote

BACKGROUND: Genistein and daidzein are two major soybean isoflavones. They have received increasing attention because of their possible roles for cancer prevention. However, their mechanisms of action and molecular targets on the human colon cancer cells are not fully understood. METHODS: Human colon cancer HCT-116 cells were treated with genistein and daidzein to investigate their effects on the cell growth and this was analyzed with MTT assay. TUNEL assay and Hoechst33342 stain were carried out to identify apotosis. RESULTS: Daidzein was able to inhibit cell proliferation and induce apoptosis of the HCT-116 cells, but genistein didn't affect the cell growth. The ER antagonist ICI182780 didn't attenuate the antiproliferative and proapoptotic effects of daidzein: this means the effect of daidzein on the HCT-116 cells may not be dependent on the ER pathway. The other soybean isoflavone, genistein, attenuated the effects of daidzein on the HCT-116 cells and its mechanism should be elucidated. CONCLUSIONS: These data suggest that daidzein may act as a preventive agent on human colon cancer, and its mechanism of action doesn't involve the ER-dependent pathway.
Apoptosis ; Cell Proliferation ; Colon* ; Colonic Neoplasms* ; Genistein* ; HCT116 Cells* ; Humans* ; In Situ Nick-End Labeling ; Isoflavones ; Soybeans

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*