• Journal of Animal Science and Technology
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• v.48 no.2
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• pp.191-202
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• 2006

In addition to the removal of dying or dead lutein cells by phagocytosis in many species, macrophages exert both luteotropic effect during maturation period and luteolytic effect during degenerative period via mediating autocrine/paracrine actions of self-producing cytokines in the corpus luteum. In this experiment, immunohistochemical and transmission electron microscopic (TEM) studies were performed to observe the morphologic changes of luteal macrophages during luteolysis. A small number of macrophages and low immunoreactivity were present at the mature stage. The number of macrophages and immunoreactivity gradually increased along the advance of luteolysis. Two subtypes of macrophages could be observed through TEM observation. One type of macrophage located between the large lutein cells contained no lipid droplets in their cytoplasm at mature stage. The other type of macrophage located near the blood vessels contained many lipid droplets in their cytoplasm during luteolysis. Particularly, no phagocytic macrophages were observed, which suggested the macrophages in the porcine corpus luteum did not involve in the phagocytotic elimination of dying lutein cells.

• Radiation Oncology Journal
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• v.18 no.1
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• pp.60-67
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• 2000

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 matatonin treatment. To analysis crypt survival, microcolony survival assay was done according to Withers and 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 its protective effect seems not to be related with apoptosis protection effect.

• Radiation Oncology Journal
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• v.25 no.4
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• pp.227-232
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• 2007

Purpose: To investigate the flavopiridol effect on radiation-induced apoptosis and expression of apoptosisrelated genes of human laryngeal and lung cancer cells. Materials and Methods: A human laryngeal cancer cell line, AMC-HN3 and a human lung cancer cell line, NCI-H460, were used in the study. The cells were divided into four groups according to the type of treatment: 1) control groups; 2) cells that were only irradiated; 3) cells treated only with flavopiridol; 4) cells treated with flavopiridol and radiation simultaneously. The cells were irradiated with 10 Gy of X-rays using a 4 MV linear accelerator. Flavopiridol was administered to the media at a concentration of 100 nM for 24 hours. We compared the fraction of apoptotic cells of each group 24 hours after the initiation of treatment. The fraction of apoptotic cells was detected by measurement of the sub-G1 fractions from a flow cytometric analysis. The expression of apoptosis-regulating genes, including cleaved caspase-3, cleaved PARP (poly (ADP-ribose) polymerase), p53, p21, cyclin D1, and phosphorylated Akt (protein kinase B) were analyzed by Western blotting. Results: The sub-G1 fraction of cells was significantly increased in the combination treatment group, as compared to cells exposed to radiation alone or flavopiridol alone. Western blotting also showed an increased expression of cleaved caspase-3 and cleaved PARP expression in cells of the combination treatment group, as compared with cells exposed to radiation alone or flavopiridol alone. Treatment with flavopiridol down regulated cyclin 01 expression of both cell lines but its effect on p53 and p21 expression was different according to each individual cell line. Flavopiridol did not affect the expression of phophorylated Akt in both cell lines. Conclusion: Treatment with flavopiridol increased radiation-induced apoptosis of both the human laryngeal and lung cancer cell lines. Flavopiridol effects on p53 and p21 expression were different according to the individual cell line and it did not affect Akt activation of both cell lines.

• Radiation Oncology Journal
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• v.22 no.2
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• pp.145-154
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• 2004

Purpose : To examine whether a synthetic bile acid derivatives (HS-1200) sensitizes the radiation-induced apoptosis in human breast cancer cells (MCF-7) and to investigate the underlying mechanism. Materials and Methods : Human breast cancer cells (MCF-7) in exponential growth phase were treated with HS-1200 for 24 hours at 37$^{\circ}C$ with 5$\%$ CO$_{2}$ in air atmosphere. After removal of HS-1200, cells were irradiated with 2$\~$8 Gy X-ray, and then cultured Ii drug-free media for 24-96 hours. The effect of radiation on the clonogenicity of MCF-7 cells was determined with clonogenic cell survival assay with 16$\mu$M of HS-1200. The induction of apoptosis was determined using agarose gel electrophoresis and Hoechst staining. The expression level of apoptosis-related molecules, such as PARP, Bax, Bcl-2, Bak and AIF, were assayed by Western blotting analysis with 40$\mu$M of HS-1200 combined with 8 Gy irradiation. To examine the cellular location of cytochrome c, bax and AIF immunofluorescent stainings were undertaken. Results : Treatment of MCF-7 cells with 40$\mu$M of HS-1200 combined with 8 Gy irradiation showed several changes associated with enhanced apoptosis by agarose gel electrophoresis and Hoechst staining. HS-1200 combined with 8 Gy irradiation treatment also enhanced production of PARP cleavage products and increased Bax/Bcl-2 ratio by Western blotting. Loss of mitochondrial membrane potential ($\Delta$$\psi$$_{m}$) and increased cytochrome c staining indicated that cytochrome c had been released from the mitochondria in HS-1200 treated cells. Conclusion : We demonstrated that combination treatment with a synthetic chenodeoxycholic acid derivative HS-1200 and irradiation enhanced radiation-induced apoptosis of human breast cancer cells (MCF-7). We suggest that the increased Bax/Bcl-2 ratio In HS-1200 co-treatment group underlies the increased radio sensitivity of MCF-7 cells. Further futures studies are remained elusive.

• Korean Journal of Veterinary Research
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• v.41 no.2
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• pp.203-210
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• 2001

Here, we compared the effectiveness of 50 MeV($p{\to}RBe^+$) cyclotron fast neutrons versus $^{60}Co$ ${\gamma}$-rays by the apoptotic fragment frequency in both rat peripheral lymphocytes and crypt cells to check a radiobiological endpoint. The incidence of apoptotic cell death was increased in all irradiated groups, and radiation at all doses trigger rapid changes in both crypt cells and peripheral lymphocytes. These data suggest that apoptosis may play an important role in homeostasis of damaged radiosensitive target organ by removing damaged cells. The curve of dose-effect relationship for these data of apoptotic fragments frequencies was $y=0.3+(6.512{\pm}0.279)D(r^2=0.975)$ after neutrons, while $y=0.3+(4.435{\pm}0.473)D+(-1.300{\pm}0.551)D^2(r^2=0.988)$ after ${\gamma}$-rays. In addition, $y=3.5+(118.410{\pm}10.325)D+(-33.548{\pm}12.023)D^2(r^2=0.992)$ after ${\gamma}$-rays in rat lymphocytes. A significant dose-response relationship was found between the frequency of apoptotic cell and dose. These data show a trend towards increase of the numbers of apoptotic cells with increasing dose. Dose-response curves for high and low linear energy transfer (LET) radiation modalities in these studies were different. The relative biological effectiveness (RBE) value for crypt cells was 1.919. In addition, there were significant peaks on apoptosis induction at 4 and 6h after irradiation, and the morphological findings of the irradiated groups were typical apoptotic fragments in crypt cells that were hardly observed in the control group. Thus, apoptosis induction in both crypt cells and peripheral lymphocytes could be a useful endpoint of rat model for studying screening test and microdosimetic indicator to evaluate the biological effects of radiation-induced cell damage.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.329-336
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• 2017

This study examined the effects of polyphenols in grape pruning stem extracts (GPSE) using grape stems discarded after harvest. The inhibitory effects on allergy, proliferation, and apoptosis in UVB-induced HR-1 hairless mice were analyzed. The applicability as a material for functional food and functional cosmetics was evaluated. The contents of the active ingredients of GPSE were analyzed by HPLC. After UVB irradiation on the dermis of HR-1 hairless mice, the number of mast cells was determined by toluidine blue staining to confirm that the skin allergic reaction was caused by GPSE. The results were similar to the normal group in the group applied GPSE, and there was no allergic reaction in the GPSE application group and a significant decreased compared to the sun cream control. PCNA immunohistochemical staining of the epidermal proliferation factor had an inhibitory effect on epidermal epithelial cell proliferation in all concentrations of GPSE and serum base mixture as an average of 42% compared to the control group. The mixture of GPSE and serum base suppressed the apoptosis inhibition rate by 27% on average compared to the control. These results confirmed the inhibitory effects GPSE on the allergic, proliferation, and apoptosis activities by with a serum base on UVB-induced skin damage. GPSE is a functional ingredient with potential skin protection effects, and has a high utilization as an ingredient for functional cosmetics.

• Radiation Oncology Journal
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• v.27 no.4
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• pp.218-227
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• 2009

Purpose: To investigate the radiobiologic effects of neutron and X-ray irradiation on DU-145 prostate carcinoma cells by identifying the differences of HIF-$1\alpha$ expression and apoptosis. Materials and Methods: Nude mice were injected with the human prostate cancer cell line, DU-145, and then irradiated with 2 Gy and 10 Gy X-rays, or 0.6 Gy and 3.3 Gy neutrons, respectively. The mice were sacrificed at 24 hours and 120 hours after irradiation. The expression levels of HIF-$1\alpha$, Bcl-2 and Bax were compared with immunohistochemical staining and western blotting. The apoptotic indexes were compared with the Terminal deoxynucleotidyl biotin-dUTP nick and labeling (TUNEL) assay. Results: At day 1, HIF-$1\alpha$ and Bcl-2 expression decreased, while Bax expression and the number of TUNEL positive cells increased in neutron irradiated groups for the control and X-ray irradiated groups. The Bcl-2/Bax ratio was significantly lower in the neutron irradiated groups regardless of dose (p=0.001). The same pattern of the differences in the expressions of the HIF-$1\alpha$, Bcl-2, Bax, Bcl-2/Bax ratio, and apoptotic indexes were indentified at day 5. HIF-$1\alpha$ expression was related with Bcl-2 (p=0.031), Bax (p=0.037) expressions and the apoptotic indexes (p=0.016) at day 5. Conclusion: Neutron irradiation showed a decrease in HIF-$1\alpha$, Bcl-2 expression, and Bcl-2/Bax ratio, but increased Bax expression regardless of dose. This study suggests that the differences radiobiological responses between photon and neutron irradiation may be related to different HIF-$1\alpha$ expression and subsequent apoptotic protein expressions.

• Applied Microscopy
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• v.26 no.3
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• pp.369-377
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• 1996

Ultrastructural changes of chloride cells in the gill from seawater-adapted killifish (Orizias latipes) were studied with a transmission electronmicroscope. Chloride cells contain many mitochondria and specifically developed tubular systems. Apical pits, formed by several neighboring chloride cells, were exposed to the environment. Degeneration and death of the chloride cells by apoptosis occurred more frequently than by necrosis. Apoptotic chloride cells shrank, became to apoptotic bodies, and eventually were phagocytosed and digested by the microphages around them. We conclude that apoptosis plays an important role in increased cellular turnover of chloride cells for the osmoregulation caused by changes in salinity of the environment.

• Proceedings of the Korean Information Science Society Conference
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• 2011.06c
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• pp.184-187
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• 2011

셀 트래킹의 목적은 셀의 이동(translocation), 분할(mitosis), 통합(fusion), 아포토시스(apoptosis), 셀의 모양 변형, 셀들 간의 상호 작용 등을 포함하는 모든 셀의 행동들을 분석하기 위한 것이다. 셀은 시간이 경과함에 따라 새롭게 나타나기도, 죽기도 하며 한 개 이상의 셀이 부분적으로 겹쳐 클러스터를 형성하기도 하고 클러스터는 다시 여러 개의 셀로 분리되기도 한다. 본 연구에서는 현미경으로부터 얻은 이미지에서 셀 트래킹을 위한 이미지 처리 방법과 오목 정점을 이용하여 셀과 클러스터를 구분하여 계수하는 방법을 제시한다. 또한 타원 근사법(ellipse fitting)을 통해 클러스터를 몇 개의 셀로 분리하기 위한 방법을 제시하고 결과를 분석한다.

• Korean Journal of Veterinary Research
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• v.41 no.4
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• pp.571-578
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• 2001

To evaluate if the apoptotic fragment assay could be used to estimate the dose prediction after radiation exposure, we examined apoptotic mouse crypt cells per 1,000 cells after whole body $^{60}Co$ $\gamma$-rays and 50MeV ($p{\rightarrow}Be^+$) cyclotron fast neutron irradiation in the range of 0.25 to 1 Gy, respectively. The incidence of apoptotic cell death rose steeply at very low doses up to 1 Gy, and radiation at all doses tigger rapid changes in crypt cells in stem cell region. These data suggest that apoptosis may play an important role in homeostasis of damaged radiosensitive target organ by removing damaged cells. The curve of dose-effect relationship for the data of apoptotic fragments was obtained by the linear-quadratic model $y=0.18+(9.728{\pm}0.887)D+(-4.727{\pm}1.033)D^2$ ($r^2=0.984$) after $\gamma$-rays irradiation, while $y=0.18+(5.125{\pm}0.601)D+(-2.652{\pm}0.7000)D^2$ ($r^2=0.970$) after neutrons in mice. The dose-response curves were linear-quadratic, and a significant dose-response relationship was found between the frequency of apoptotic cell and dose. These data show a trend towards increase of the numbers of apoptotic crypt cells with increasing dose. Both the time course and the radiation dose-response curve for high and low linear energy transfer (LET) radiation modalities were similar. The relative biological effectiveness (RBE) value for crypt cells was 2.072. In addition, there were significant peaks on apoptosis induction at 4 and 6h after irradiation, and the morpholoigcal findings of the irradiated groups were typical apoptotic fragments in crypt cells that were hardly observed in the control group. Thus, apoptosis in crypt cells could be a useful in vivo model for studying radio-protective drug sensitivity or screening test, microdosimetric indicator and radiation-induced target organ injury. Since the apoptotic fragment assay is simple, rapid and reproducible in the range of 0.25 to 1 Gy, it will also be a good tool for evaluating the dose response of radiation-induced organ damage in vivo and provide a potentially valuable biodosimetry for the early dose prediction after accidental exposure.