• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.287-294
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• 2011

Autophagy is a process of intracellular bulk protein degradation, in which the accumulated proteins and cytoplasmic organelles are degraded. It plays important roles in cellular homeostasis, apoptosis, and development, but its role during early embryo development remains contentious. Therefore, in the present study, we investigated the effects of 3-methyladenine (3-MA) on early embryonic development in pigs, we also investigated several indicators of developmental potential, including mitochondrial distribution, genes expressions (autophagy-, apoptosis- related genes), apoptosis and ER-stress, which are affected by 3-MA. After in vitro maturation and fertilization, presumptive pig embryos were cultured in PZM-3 medium supplemented with 3-MA for 2 days at $39^{\circ}C$ 5% $CO_2$ in air. Developmental competence to the blastocyst stage in the presence of 3-MA was gradually decreased according to increasing concentration. Thus, all further experiments were performed using 2 mM 3-MA. Blastocysts that developed in the 3-MA treated group decreased LC3-II intensity and expressions of autophagy related genes than those of the untreated control, resulting in down-regulates the autophagy. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 3-MA treated group compared with control ($6.0{\pm}1.0$ vs $3.3{\pm}0.6$, p<0.05). Also, the expression of the pro-apoptotic gene Bax increased in 3-MA treated group, whereas expression of the anti-apoptotic gene Bcl-XL decreased. Mito Tracker Green FM staining showed that blastocysts derived from the 3-MA treated group had lower mitochondrial integrity than that of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. Then, the expression of the spliced form of pXBP-1 product (pXBP-1s) increased in 3-MA treated group, resulting increase of ER-stress. Taken together, these results indicate that inhibition of autophagy by 3-MA is closely associated with apoptosis and ER-stress during preimplantation periods of porcine embryos.

• The Journal of the Korean bone and joint tumor society
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• v.14 no.2
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• pp.106-118
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• 2008

Purpose: Disturbed cell cycle regulatory proteins are key events underlying the development and/or progression of human malignancies. The aim of this study is to evaluate the protein expression status involved in G1/S cell cycle in human soft tissue sarcoma. Materials and Methods: We simultaneously evaluated the expression of Cyclin D1, Cyclin E, CDK4, CDK2, p16, p27, Rb, E2F1, p53 and Ki-67 by immunohistochemistry in 43 cases of soft tissue sarcoma Results: The Cyclin D1, Cyclin E, CDK4, CDK2, E2F1, and p53 were expressed in 25 (58.1%), 18 (41.9%), 13 (30.2%), 33 (76.7%), 20 (46.5%), and 18 cases (41.9%). The p16, p27, and Rb expressions were decreased in 26 (60.5%), 22 (51.2%) and 19 cases (44.2%). All of the cases showed alterations of more than one out of the above proteins. The increased Cyclin E expression and Ki-67 labeling index (LI) were significantly associated with histologic grade. The Cyclin E and E2F-1 expressions were increased in relapsed cases and the CDK4 expression was increased in cases of metastasis. Conclusion: Alterations of G1/S cell cycle regulatory proteins may play an important role in the tumoriogensis of soft tissue sarcomas. Our results suggest that increased expressions of Cyclin E, E2F1, and CDK4 were associated with tumor relapse or metastasis and could be considered as parameters of prognosis of soft tissue sarcoma.

• Molecules and Cells
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• v.42 no.6
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• pp.448-459
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• 2019

The phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) signaling pathway is a promising target for gastric cancer (GC) treatment; however the efficacy of PI3K/mTOR dual inhibitors in GC has not yet been maximized. Additionally, the effect of autophagy regulation by PI3K/mTOR dual inhibitors has not been clearly elucidated in GC treatment. We aimed to show that our newly developed PI3K/mTOR dual inhibitor, CMG002, when combined with an autophagy inhibitor, chloroquine (CQ), potently induces effective cancer cell death in Epstein-Barr virus (EBV)-associated gastric cancer (EBVaGC) cells, where both the PI3K/AKT/mTOR and autophagy pathways play important roles in disease pathogenesis. EBV- and mock-infected AGS and NUGC3 GC cell lines were treated with CMG002 +/- CQ. PI3K/AKT/mTOR signaling pathway mediators, cellular apoptosis and autophagy markers were confirmed by Western blot assay. Cell viability was assessed using the Cell Counting Kit-8 (CCK-8) assay. CMG002 effectively blocked the PI3K/AKT/mTOR pathway by markedly decreasing phosphorylation of AKT and its downstream mediator S6. CMG002 induced G0/G1 cell cycle arrest and enhanced apoptotic cell death in AGS and NUGC3 cells, particularly EBV-infected cells compared with mock-infected cells, as confirmed by flow cytometric analyses and TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) assays. The combination of CMG002 plus CQ synergistically increased apoptotic cell death in EBV-infected GC cell lines when compared with CMG002 alone (P < 0.05). Our results suggest that the new PI3K/mTOR dual inhibitor, CMG002, when used in combination with the autophagy inhibitor, CQ, provides enhanced therapeutic efficacy against EBVaGC.

• The Korean Journal of Physiology and Pharmacology
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• v.25 no.2
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• pp.147-157
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• 2021

Coronary microembolization (CME) is associated with cardiomyocyte apoptosis and cardiac dysfunction. Puerarin confers protection against multiple cardiovascular diseases, but its effects and specific mechanisms on CME are not fully known. Hence, our study investigated whether puerarin pretreatment could alleviate cardiomyocyte apoptosis and improve cardiac function following CME. The molecular mechanism associated was also explored. A total of 48 Sprague-Dawley rats were randomly divided into CME, CME + Puerarin (CME + Pue), sham, and sham + Puerarin (sham + Pue) groups (with 12 rats per group). A CME model was established in CME and CME + Pue groups by injecting 42 ㎛ microspheres into the left ventricle of rats. Rats in the CME + Pue and sham + Pue groups were intraperitoneally injected with puerarin at 120 mg/kg daily for 7 days before operation. Cardiac function, myocardial histopathology, and cardiomyocyte apoptosis index were determined via cardiac ultrasound, hematoxylin-eosin (H&E) and hematoxylin-basic fuchsin-picric acid (HBFP) stainings, and TdT-mediated dUTP nick-end labeling (TUNEL) staining, respectively. Western blotting was used to measure protein expression related to the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β) pathway. We found that, puerarin significantly ameliorated cardiac dysfunction after CME, attenuated myocardial infarct size, and reduced myocardial apoptotic index. Besides, puerarin inhibited cardiomyocyte apoptosis, as revealed by decreased Bax and cleaved caspase-3, and up-regulated Bcl-2 and PI3K/Akt/GSK-3β pathway related proteins. Collectively, puerarin can inhibit cardiomyocyte apoptosis and thus attenuate myocardial injury caused by CME. Mechanistically, these effects may be achieved through activation of the PI3K/Akt/GSK-3β pathway.

• Korean Journal of Food Science and Technology
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• v.44 no.4
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• pp.452-459
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• 2012

This study was conducted in order to investigate the effect of resveratrol on the induction of apoptosis in MDA-MB-231 breast cancer cells. The result of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl terazolium bromide (MTT) assay shows that cell viability significantly decreased in a dose and time-dependent manner. 4',6-diamidino-2-phenylindole (DAPI) staining shows significantly increased chromatin condensation in a dose and time-dependent manner. Resveratrol increased the expression of p53, cleaved-caspase-3, and cleaved-caspase-9, whereas the expression of PI3K/Akt decreased in a time-dependent manner. We investigated the in vivo tumor growth inhibitory effect of resveratrol. Tumor volume was significantly decreased in the 50 mg/kg resveratrol-administration group compared to the control group. In the 50 mg/kg treated group. Apoptosis cells were frequently observed by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay. Immunohistochemistry staining shows increased the expression of p53, cytochrome-C, and cleaved-caspase-3 in the 50 mg/kg treated group. These results indicate that resveratrol induced apoptosis through PI3K/Akt and p53 signal pathway in MDA-MB-231 cell.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.18 no.10
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• pp.707-716
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• 2017

The advances in image display devices necessitate display images suitable for the user's purpose. The display devices should be able to provide object-based image information when a depthmap is required. In this paper, we represent the algorithm using a histogram-based image segmentation method for depthmap generation. In the conventional K-means clustering algorithm, the number of centroids is parameterized, so existing K-means algorithms cannot adaptively determine the number of clusters. Further, the problem of K-means algorithm tends to sink into the local minima, which causes over-segmentation. On the other hand, the proposed algorithm is adaptively able to select centroids and can stand on the basis of the histogram-based algorithm considering the amount of computational complexity. It is designed to show object-based results by preventing the existing algorithm from falling into the local minimum point. Finally, we remove the over-segmentation components through connected-component labeling algorithm. The results of proposed algorithm show object-based results and better segmentation results of 0.017 and 0.051, compared to the benchmark method in terms of Probabilistic Rand Index(PRI) and Segmentation Covering(SC), respectively.

• Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
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• v.31 no.4
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• pp.331-339
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• 2013

LiDAR has been one of the widely used and important technologies for 3D modeling of ground surface and objects because of its ability to provide dense and accurate range measurement. The objective of this research is to develop a method for automatic detection and modeling of railroad power lines using high density LiDAR data and RANSAC algorithms. For detecting railroad power lines, multi-echoes properties of laser data and shape knowledge of railroad power lines were employed. Cuboid analysis for detecting seed line segments, tracking lines, connecting and labeling are the main processes. For modeling railroad power lines, iterative RANSAC and least square adjustment were carried out to estimate the lines parameters. The validation of the result is very challenging due to the difficulties in determining the actual references on the ground surface. Standard deviations of 8cm and 5cm for x-y and z coordinates, respectively are satisfactory outcomes. In case of completeness, the result of visual inspection shows that all the lines are detected and modeled well as compare with the original point clouds. The overall processes are fully automated and the methods manage any state of railroad wires efficiently.

• Journal of Korea Water Resources Association
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• v.55 no.spc1
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• pp.1177-1185
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• 2022

The water and sewage system is an infrastructure that provides safe and clean water to people. In particular, since the water and sewage pipelines are buried underground, it is very difficult to detect system defects. For this reason, the diagnosis of pipelines is limited to post-defect detection, such as system diagnosis based on the images taken after taking pictures and videos with cameras and drones inside the pipelines. Therefore, real-time detection technology of pipelines is required. Recently, pipeline diagnosis technology using advanced equipment and artificial intelligence techniques is being developed, but AI-based defect detection technology requires a variety of learning data because the types and numbers of defect data affect the detection performance. Therefore, in this study, various defect scenarios are implemented using 3D printing model to improve the detection performance when detecting defects in pipelines. Afterwards, the collected images are performed to pre-processing such as classification according to the degree of risk and labeling of objects, and real-time defect detection is performed. The proposed technique can provide real-time feedback in the pipeline defect detection process, and it would be minimizing the possibility of missing diagnoses and improve the existing water and sewerage pipe diagnosis processing capability.

• Journal of Veterinary Science
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• v.23 no.2
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• pp.40.1-40.13
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• 2022

Background: Somatic cell nuclear transfer (SCNT) is used widely in cloning, stem cell research, and regenerative medicine. The type of donor cells is a key factor affecting the SCNT efficiency. Objectives: This study examined whether urine-derived somatic cells could be used as donors for SCNT in pigs. Methods: The viability of cells isolated from urine was assessed using trypan blue and propidium iodide staining. The H3K9me3/H3K27me3 level of the cells was analyzed by immunofluorescence. The in vitro developmental ability of SCNT embryos was evaluated by the blastocyst rate and the expression levels of the core pluripotency factor. Blastocyst cell apoptosis was examined using a terminal deoxynucleotidyl transferase dUTP nick end-labeling assay. The in vivo developmental ability of SCNT embryos was evaluated after embryo transfer. Results: Most sow urine-derived cells were viable and could be cultured and propagated easily. On the other hand, most of the somatic cells isolated from the boar urine exhibited poor cellular activity. The in vitro development efficiency between the embryos produced by SCNT using porcine embryonic fibroblasts (PEFs) and urine-derived cells were similar. Moreover, The H3K9me3 in SCNT embryos produced from sow urine-derived cells and PEFs at the four-cell stage showed similar intensity. The levels of Oct4, Nanog, and Sox2 expression in blastocysts were similar in the two groups. Furthermore, there is a similar apoptotic level of cloned embryos produced by the two types of cells. Finally, the full-term development ability of the cloned embryos was evaluated, and the cloned fetuses from the urine-derived cells showed absorption. Conclusions: Sow urine-derived cells could be used to produce SCNT embryos.

• Journal of Pharmacopuncture
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• v.4 no.1
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• pp.123-124
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• 2001

Purpose. Free radicals are implicated in the pathophysiology of aging, ischemic injury and neurodegenerative disorders. To deform]no whether Hominis Placenta extract prevents $H_2O_2$-induced apoptosis, we have performed morphological and biochemical analyses for the detection of apoptotic phenomena in the pineal tumor cell line $PGT-{\beta}$ We have also peformed cytochemical and immunocytochemical analyses for the detection of changes in nitric oxide synthase (NOS) activity and estimated the expression . of apoptotic genes using reverse transcription-polymerase chain reaction (RT-PCR) Methods. $PGT-{\beta}\;cells$ were pretreated with Hominis Placenta extracts $(0,\;10^{-2}\;{\mu}g/ml)$ for 2 hours and then exposed to $H_2O_2\;(0,\;50\;{\mu}M)$ for 3 hours. Appearance of apoptotic characteristics were monitored using 4, 6-diamidino-2-phenylindole dihydrochloride (DAPI) staining assay, terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) assay and flow cytometric analysis. NOS activity was measured by NADPH-diaphorase cytochemistry. Expression of inducible NOS (iNOS) and nuclear factor kappa B (NF k B) was assessed via immunocytochemistry. The expression of apoptotic genes was examined by RT-PCR. Results. After 3 flours of exposure to $H_2O_2$, it was shown that $PGT-{\beta}\;cells$ treated with $H_2O_2(50\;{\mu}M)$ exhibit classical apoptotic features and increases in NOS activity and caspase-3 expression. Treatment with Hominis Placenta extract resulted in a reduced occurrence of apoptotic features. DAPI staining, TUNEL and flow cytometric assays revealed decreases in the occurrence of nuclear fragmentation and in the sub-Gl fraction in the $PGT-{\beta}\;cells$ treated with Hominis Placenta extract. Cells treated with Hominis Placenta extract also showed lower activity of NADPH-diaphorase and immunoreactivities of both iNOS and NF k B than those of $H_2O_2$-treated cells which were not treated with Hominis Placenta extract. By RT-PCR, it was shown that the level of caspase-3 mRNA was derreased In the cells treated with Hominis Placenta . extract. Conclusions. This study shows that Hominis Placenta extract prevents $H_2O_2$-induced apoptosis in $PGT-{\beta}\;cells$; inhibitions of iNOS and caspnse-3 are possible mechanisms of the protection against apoptosis.