• Title/Summary/Keyword: Cycle-Dependent

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Walnut phenolic extracts reduce telomere length and telomerase activity in a colon cancer stem cell model

  • Shin, Phil-Kyung;Zoh, Yoonchae;Choi, Jina;Kim, Myung-Sunny;Kim, Yuri;Choi, Sang-Woon
    • Nutrition Research and Practice
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    • v.13 no.1
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    • pp.58-63
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    • 2019
  • BACKGROUND/OBJECTIVES: Telomeres are located at the chromosomal ends and progressively shortened during each cell cycle. Telomerase, which is regulated by hTERT and c-MYC, maintains telomeric DNA sequences. Especially, telomerase is active in cancer and stem cells to maintain telomere length for replicative immortality. Recently we reported that walnut phenolic extract (WPE) can reduce cell viability in a colon cancer stem cell (CSC) model. We, therefore, investigated the effect of WPE on telomere maintenance in the same model. MATERIALS AND METHODS: $CD133^+CD44^+$ cells from HCT116, a human colon cancer cell line, were sorted by Fluorescence-activated cell sorting (FACS) and treated with WPE at the concentrations of 0, 10, 20, and $40{\mu}g/mL$ for 6 days. Telomere lengths were assessed by quantitative real-time PCR (qRT-PCR) using telomere specific primers and DNA extracted from the cells, which was further adjusted with single-copy gene and reference DNA ($ddC_t$). Telomerase activity was also measured by qRT-PCR after incubating the PCR mixture with cell protein extracts, which was adjusted with reference DNA ($dC_t$). Transcriptions of hTERT and c-MYC were determined using conventional RT-PCR. RESULTS: Telomere length of WPE-treated cells was significantly decreased in a dose-dependent manner ($5.16{\pm}0.13$ at $0{\mu}g/mL$, $4.79{\pm}0.12$ at $10{\mu}g/mL$, $3.24{\pm}0.08$ at $20{\mu}g/mL$ and $3.99{\pm}0.09$ at $40{\mu}g/mL$; P = 0.0276). Telomerase activities concurrently decreased with telomere length ($1.47{\pm}0.04$, $1.09{\pm}0.01$, $0.76{\pm}0.08$, and $0.88{\pm}0.06$; P = 0.0067). There was a positive correlation between telomere length and telomerase activity (r = 0.9090; P < 0.0001). Transcriptions of both hTERT and c-MYC were also significantly decreased in the same manner. CONCLUSION: In the present cell culture model, WPE reduced telomere maintenance, which may provide a mechanistic link to the effect of walnuts on the viability of colon CSCs.

Ginsenoside 20(S)-protopanaxadiol induces cell death in human endometrial cancer cells via apoptosis

  • Jo, Hantae;Jang, Dongmin;Park, Sun Kyu;Lee, Mi-Gi;Cha, Byungsun;Park, Chaewon;Shin, Yong Sub;Park, Hyein;Baek, Jin-myoung;Heo, Hyojin;Brito, Sofia;Hwan, Hyun Gyu;Chae, Sehyun;Yan, Shao-wei;Lee, Changho;Min, Churl K.;Bin, Bum-Ho
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.126-133
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    • 2021
  • Background: 20(S)-protopanaxadiol (20(S)-PPD), one of the aglycone derivatives of major ginsenosides, has been shown to have an anticancer activity toward a variety of cancers. This study was initiated with an attempt to evaluate its anti-cancer activity toward human endometrial cancer by cell and xenograft mouse models. Methods: Human endometrial cancer (HEC)-1A cells were incubated with different 20(S)-PPD concentrations. 20(S)-PPD cytotoxicity was evaluated using MTT assay. Apoptosis was detected using the annexin V binding assay and cell cycle analysis. Cleaved poly (ADP-ribose) polymerase (PARP) and activated caspase-9 were assessed using western blotting. HEC-1A cell tumor xenografts in athymic mice were generated by inoculating HEC-1A cells into the flank of BALB/c female mice and explored to validate 20(S)-PPD anti-endometrial cancer toxicity. Results: 20(S)-PPD inhibited HEC-1A cell proliferation in a dose-dependent manner with an IC50 value of 3.5 μM at 24 h. HEC-1A cells morphologically changed after 20(S)-PPD treatment, bearing resemblance to Taxol-treated cells. Annexin V-positive cell percentages were 0%, 10.8%, and 58.1% in HEC-1A cells when treated with 0, 2.5, and 5 μM of 20(S)-PPD, respectively, for 24 h. 20(S)-PPD subcutaneously injected into the HEC-1A cell xenograft-bearing mice three times a week for 17 days manifested tumor growth inhibition by as much as 18% at a dose of 80 mg/kg, which sharply contrasted to controls that showed an approximately 2.4-fold tumor volume increase. These events paralleled caspase-9 activation and PARP cleavage. Conclusion: 20(S)-PPD inhibits endometrial cancer cell proliferation by inducing cell death via a caspase-mediated apoptosis pathway. Therefore, the 20(S)-PPD-like ginsenosides are endowed with ample structural information that could be utilized to develop other ginsenoside-based anticancer agents.

Anti-cancer effects of Salicornia herbacea extract in OVCAR-3 cells by induction of apoptosis pathway (함초 추출물의 세포사멸 유도에 의한 난소암세포에서 항암 효과)

  • La, Yu Ri;Lee, You Rim;Lee, Dong Seok;Kim, Soo Hwan;Lee, Hyeong Seon
    • Korean Journal of Food Science and Technology
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    • v.53 no.1
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    • pp.34-39
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    • 2021
  • This study investigated the anti-cancer effects of Salicornia herbacea L. fractions in human ovarian cancer cells (OVCAR-3). S. herbacea powder was extracted with 95% EtOH and sequentially fractionated with hexane, dichloromethane (DCM), ethyl acetate, butanol, and H2O. Further, the growth inhibitory effects of the six fractions were determined using the MTS assay. The DCM fraction dramatically decreased cell viability. Similarly, the cell cycle was arrested at the subG1 phase in DCM-treated cells. To confirm apoptosis, the cells were stained with annexin V/FITC-PI solution. Total, early, and late apoptotic cells were significantly increased in the DCM fraction. The mRNA expression of Bcl-2 was reduced, whereas the pro-apoptotic factors Bax and Bak were increased in DCM fraction-treated cells. These results indicated that the DCM fraction in S. herbacea exhibited strong apoptotic effects through the p53-dependent signaling pathway.

Inhibition of Proliferation of LNCaP Prostate Cells by Corni Fructus Extract Is Associated with a Decrease in the Expression of Benign Prostatic Hyperplasia-Causing Factors (산수유 추출물에 의한 LNCaP 전립선 세포의 증식 억제 및 양성 전립선 비대증 유발 인자의 발현에 미치는 영향)

  • Kim, Min Yeong;Ji, Seon Yeong;Hwangbo, Hyun;Lee, Hyesook;Hong, Su Hyun;Choi, Yung Hyun
    • Journal of Korean Medicine for Obesity Research
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    • v.21 no.1
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    • pp.10-21
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    • 2021
  • Objectives: Benign prostatic hyperplasia (BPH) is a progressive pathological condition characterized by excessive proliferation of the prostate. In this study, we evaluated the effect of Corni Fructus water extract (CF) on the promotion of prostate cell proliferation by dihydrotestosterone (DHT). Methods: The effect of CF on the proliferation of LNCaP prostate cells was evaluated, and DHT was treated to induce an in vitro BPH model. To study the mechanism of inhibition of cell proliferation and BPH by CF, changes in the expression of key factors related to cell cycle and BPH were investigated. We further investigated the effect on the production of reactive oxygen species (ROS) and nitric oxide (NO) to evaluate the antioxidant and anti-inflammatory efficacy of CF. Results: Inhibition of LNCaP cell proliferation by CF was associated with decreased expression of cyclin D1 and cyclin A and increased expression of cyclin-dependent kinase inhibitor p21. CF also suppressed expression of BPH inducing factors such as 5α-reductase type 2 and androgen receptor (AR) as well as prostate specific antigen (PSA). Furthermore, CF significantly blocked DHT-induced LNCaP cell proliferation and effectively attenuated DHT-induced expression of BPH mediators and cyclins. In addition, CF inhibited DHT-induced oxidative and inflammatory reactions by inhibiting production of ROS and NO. Conclusion: Our results demonstrated that CF probably acted as 5α-reductase type 2 inhibitor, preventing the 5α-reductase type 2-AR signaling pathway, thereby reducing the conversion of testosterone to DHT and the expression of PSA, which is at least correlated with the antioxidant and anti-inflammatory activities of CF.

Ecotoxicity Evaluation of PFCs using Marine Invertebrate, Sea Urchin (Mesocentrotus nudus) (둥근성게(Mesocentrotus nudus)를 이용한 과불화화합물의 생태독성평가)

  • Choi, Hoon;Lee, Ju-Wook;Lee, Seung-Min;Jeon, Hyung-Ju;Heo, Seung;Hwang, Un-Ki
    • Journal of Marine Life Science
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    • v.6 no.2
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    • pp.80-87
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    • 2021
  • In this study, the toxic effects of PFOA and PFOS potassium salt on Mesocentrotus nudus using 10 min-fertilization rate and 48 h-normal embryogenesis were confirmed through the calculation of toxicity values such as Non-observed effective concentration, Low-observed effective concentration, and 50% of effective concentration. The case of 10 min-fertilization rate and 48 h-normal embryogenesis showed the concentration-dependent reduction pattern when exposed to PFOA and PFOS potassium salt, in tested concentration, respectively. The EC50 values of 10 min-fertilization rates for PFOA and PFOS potassium salt were 1346.43 mg/l and 536.18 mg/l, respectively, and the EC50 values of 48 h-normal embryogenesis were 42.67 mg/l and 17.81 mg/l, respectively. Both toxicity test methods showed high toxicity sensitivity to PFOS potassium salt. Recent studies have shown that the concentration of PFOA and PFOS in the marine environment has continuously decreased, and it is not enough to show acute toxicity to sea urchin. However, PFOA and PFOS have a very long half-life and can accumulate throughout the life of marine life, so it is still observed at a high concentration in shellfish. Therefore, a study on chronic toxicity through the whole-life cycle of marine organisms in coastal environments should be needed.

Development of Optimal Antiviral Coating Method for the Air Filtration System of Subway Station (지하역사 승강장 공조 시스템 필터용 항바이러스 코팅 성능 및 재생 성능 평가)

  • Park, Dae Hoon;Hwang, Jungho;Shin, Dongho;Kim, Younghun;Lee, Gunhee;Park, Inyong;Kim, Sang Bok;Hong, Keejung;Han, Bangwoo
    • Particle and aerosol research
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    • v.18 no.1
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    • pp.9-21
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    • 2022
  • In this study, a novel antiviral coating method for the air filtration system of subway station was investigated. Using dry aerosol coating process, we developed a high-performance antiviral air filter with spark discharger and carbon brush type ionizer. Silver nanoparticles were produced by a spark discharge generation system with ion injection system and were used as antiviral agents coated onto a medium grade air filter. The pressure drop, filtration efficiency, and antiviral ability of the filter against aerosolized MS2 virus particles as a surrogate of SARS-CoV-2 virus were tested with dust contamination. Dust contamination caused the increase of the filtration efficiency and pressure drop, while the antiviral agents (in this study, silver nanoparticles) coating did not have any significant effect on the filtration efficiency and pressure drop. Using these properties, we suggested a novel method to maximize the antiviral performance of the antiviral air filter that was contaminated by dust particles. Moreover theoretical analysis of antiviral ability with dust contamination and re-coated antiviral agents was carried out using a mathematical model to calculate the time-dependent antiviral effect of the filter under actual conditions of subway station. Our model can be used to apply on antiviral air filtration system of subway station for prevention of pandemic diffusion, and predict the life cycle of an antiviral filter.

Bioactivity enhancement of zirconia substrate by surface coating of diopside bioceramics using sol-gel method (솔젤법에 의한 다이옵사이드 생체 세라믹의 표면코팅 및 지르코니아 기판의 생체활성 증진)

  • Park, Hyunjung;Lee, Jong Kook
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.32 no.5
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    • pp.183-190
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    • 2022
  • Diopside (CaMgSi2O6) is known to have high bioactivity as well as excellent mechanical properties. In this study, we tried to improve the bioactivity of zirconia ceramics by surface coating of diopside and its bioactivity was investigated through an in vitro test. Surface coating on zirconia substrate was prepared by sol-gel method using a diopside sol which was prepared by dissolving Ca(NO3)2·4H2O, MgCl2·6H2O and Si(OC2H5)4 in ethanol with a fixed molar ratio and then hydrolysis. To examine the bioactivity of diopside coating, we examined the surface dissolution and the precipitation of new hydroxyapatite particles through in vitro test in SBF (Simulated Body Fluid) solution. Dense and thick diopside coating layers could be fabricated on zirconia substrate by sol-gel method. Also, we confirmed that they contained high bioactivity from the in vitro test, indicated the precipitation of hydroxyapatite particles after the 14 days immersion in SBF solution. In addition, we checked that the bioactivity of diopside coated layers was dependent on the repeated coating cycle and coating thickness.

The Effect of Oocyte Activation on Development of Porcine Cloned Embryos

  • Kim, Y.S.;Lee, S. L.;Park, G. J.;S. Y. Choe
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.124-124
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    • 2003
  • The successful development of embryos cloned by nuclear transfer (NT)have been dependent on a wide range of known factors including cell cycle of donor and recipient ooplast, oocyte quality, NT procedure and oocyte activation. The present study compared the development of cloned porcine embryos following different activation treatments. Cumulus-oocyte complexes (COCs) were aspirated from 26 mm follicles of slaughterhouse ovaries and cultured for 22 h in NCSU #23 medium supplemented with 10% porcine follicular fluid, 0.57 mM cysteine, 0.5 g/mL LH, 0.5 g/mL FSH and 10 ng/mL EGF. The COCs were further cultured for an additional 22 h in the same medium at $39{\cird}C$ in an atmosphere of 5% $CO_2$ in air, without hormonal supplements. Primary cultures of fibroblasts isolated from a female fetus on day 40 of gestation were established in DMEM + 15% FCS. For nuclear donation, cells at the 5th-6th passage were cultured in DMEM +0.5% FCS for 5 days in order to arrest the cells in G0/Gl. After enucleation, oocytes were reconstructed by transfer of donor cells and fusion with three DC pulses (1.4 KV/cm, 30 sec) in 0.28 M mannitol containing 0.01 mM $CaCl_2$ and 0.01 mM $MgCl_2$. Eggs were then divided into three treatment groups, control (without further treatment, Group 1), eggs cultured in 10 g/ml cycloheximide (CHX) for 5 h (Group 2), and eggs cultured in 1.9 mM 6-dimethylaminopurine (6-DMAP) for 5 h (Group 3). The eggs were then cultured in sets of 30 in 60 I drops of NCSU#23 supplemented with 4mg/ml BSA (essentially fatty acid free) until day 7 at $39{\circ}C$ in a humidified atmosphere of 5% $CO_2$. On day 4 the culture were fed by adding 20 I NCSU #23 supplemented with 10% FBS. Development rates into blastocysts were significantly higher (P<0.05) in Group 3 embryos compared to Group 1 controls ($27.6 \mu 2.7% vs. 20.1 \mu 4.1%$, respectively), but rates did not differ in Group 2 compared to control ($23.8 \mu 5.7%$). Total cell number in Group 3 blastocysts was however significantly higher (P<0.05) than in Groups 1 and 2 ($44.6 \mu 2.4 vs. 19.9 \mu 1.9 and 21.9 \mu 2.1$, respectively). These results suggest that 6-DMAP is more efficient than cycloheximide in the activation of electrically fused NT oocytes during in vitro production of cloned porcine embryos.

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The Moderating Effect of Job Satisfaction on Social Exclusion and Quality of Life of the Disabled (장애인의 사회적 배제와 삶의 질에 대한 직무만족의 조절효과)

  • Aeran Kwon;Hyeongki Lee
    • Journal of Internet of Things and Convergence
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    • v.10 no.1
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    • pp.13-20
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    • 2024
  • The purpose of this study is to objectively quantify the degree of social exclusion of the disabled and verify the impact of the derived degree of social exclusion on the quality of life of the disabled. And the purpose is to verify whether job satisfaction moderates the impact of social exclusion and quality of life of disabled people. The subjects of the study were 1,280 people extracted through a panel survey on employment of the disabled. The independent variables of social exclusion are economic difficulties, employment status, mental health, and social activities. The dependent variable was quality of life, and the control variable was job satisfaction. The research results are as follows. First, the sub-factors of social exclusion that affect quality of life were economic difficulties, mental health, participation in social activities, and employment status. Second, job satisfaction showed a moderating effect in the relationship between social exclusion and quality of life. And the sub-factors of social exclusion that have a moderating effect on quality of life were identified as economic difficulties, employment status, and social activities. In follow-up research, it will be necessary to investigate employment programs for the disabled in developed countries and analyze the implementation background, implementation details, achievements, and limitations, thereby contributing to the establishment of employment policies for the disabled. It is hoped that through this research, the vicious cycle of social exclusion and discrimination of the disabled will be improved and an opportunity will be provided to improve the quality of life through the economic activities of the disabled.

Study of the Cytotoxic Protective Effects of Jinsimyusaeng-hwan (Modified Ojayeonjong-hwan) Against Oxidative Stress (산화 스트레스에 대한 진심유생환(오자연종환 가감방) 추출물의 세포 독성 보호 효과에 관한 연구)

  • Yujin Jung;Sanghoon Hong;Myungho Jin
    • Journal of Society of Preventive Korean Medicine
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    • v.28 no.2
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    • pp.113-130
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    • 2024
  • Objectives : This study was conducted to evaluate the cytotoxic protective effects of Jinsimyusaeng-hwan (Modified Ojayeonjong-hwan) against oxidative stress. Methods : 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis-(3-ethyl-benzothiazoline-6-sulfonic acid)(ABTS) radical scavenging activity and ferric reducing ability of plasma (FRAP) method were used to estimate the antioxidant activity of Jinsimyusaeng-hwan. C2C12 myoblasts were used to reevaluate the antioxidant effects. And apoptosis analysis, mitochondrial membrane potential analysis, measurement of intracellular reactive oxygen species levels were conducted to investigate antioxidant activity of Jinsimyusaeng-hwan. Results : In comparison of DPPH free radical and ABTS cationic radical scavenging activity, it increased as the concentration of water extracts of Jinsimyusaeng-hwan(WEJ) and 70% ethanol extracts of Jinsimyusaeng-hwan (EEJ) increased. In the results of comparing the total phenol content and reducing power using the FRAP method, extracts with high total phenol content also showed high reducing power. In comparison of the protective effect against H2O2-induced oxidative stress in C2C12 myoblasts, WEJ had no significant effect, but the EEJ inhibited H2O2-mediated cytotoxicity in a concentration-dependent manner. The cytotoxic protective effect of EEJ against oxidative stress in C2C12 myoblasts was correlated with their inhibitory effects on H2O2-induced apoptosis and cell cycle arrest. In H2O2-treated C2C12 myoblasts, the apoptosis inhibitory effects of EEJ were associated with suppression of mitochondrial dysfunction and DNA damage. The protective effect of EEJ against H2O2-induced oxidative stress in C2C12 myoblasts were directly related to the inhibition of ROS generation. Conclusion : Jinsimyusaeng-hwan extracts have cytotoxic protective effects against oxidative stress, and it was better in 70% ethanol extract than in water extract.